ORCID Profile
0000-0003-2710-9324
Current Organisations
Forschungszentrum Jülich
,
Monash University
,
Murdoch University
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Publisher: Springer Science and Business Media LLC
Date: 30-09-2023
Publisher: Elsevier BV
Date: 11-2018
Abstract: Cryptosporidium species differ in host range. Parasite-host coevolution, host adaptation, and geographic segregation have led to the formation of subtype families with unique phenotypic traits within the major human-pathogenic species C. parvum and C. hominis. Transmission intensity, genetic ersity, and occurrence of genetic recombination and selective pressure have further shaped their population genetic structures. Panmixia appears to be common within the zoonotic C. parvum, especially its hypertransmissible IIaA15G2R1 subtype. Genetic recombination in C. hominis, in contrast, is more restricted to virulent subtypes, especially IbA10G2. Nonhuman primates and equine animals are commonly infected with genetically ergent C. hominis populations. Systematic studies of these and other host-adapted Cryptosporidium spp. are likely leading to improved understanding of population structures underlying various transmission patterns and intensities of Cryptosporidium.
Publisher: Elsevier BV
Date: 10-2014
DOI: 10.1016/J.EXPPARA.2014.07.004
Abstract: Little is known about the prevalence and pathogenesis of trypanosomes in Australian monotremes, and few genetic characterisation studies have been conducted with these haemoparasites. During the present investigation, molecular and microscopic methods were used to screen peripheral blood (n=28) and ectoparasites (n=10 adult ticks n=5 tick nymphs n=1 leech and n>500 tick eggs) collected from wild Tasmanian platypuses (Ornithorhynchus anatinus), for the presence of trypanosomatid-specific DNA and/or trypomastigotes. The genes for the small ribosomal subunit RNA (18S rDNA) and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) were lified and sequenced, prior to conducting phylogenetic analyses. The detection rate of the parasite-specific 18S rDNA in platypus blood was 85.7% (n=24/28), and the leech was also positive at both loci. Microscopically, high parasitaemia and the presence of abundant trypomastigotes, morphologically consistent with Trypanosoma binneyi Mackerras (1959), were observed in the blood films. Phylogenetic analyses at the 18S locus revealed the existence of four trypanosomatid-like genotypes, with variable similarity to two previously-described genotypes of T. binneyi (range of genetic p-distance: 0.0-0.5%). For the gGAPDH locus, for which only one T. binneyi sequence is available in GenBank, three genotypes closely related T. binneyi were identified (range of genetic p-distance: 0.1-0.4%). The leech-derived trypanosome isolate was virtually identical (at the two loci studied) to the other parasites sequenced from infected platypuses however, the molecular or morphological identification of the leech species was not possible. Although further studies are required, the molecular detection of trypanosomes in an aquatic leech removed from a platypus, suggests the possibility that these haematophagous hirudineans may be a vector for T. binneyi (and closely related genotypes).
Publisher: Elsevier BV
Date: 06-2008
DOI: 10.1016/J.EXPPARA.2008.02.005
Abstract: This study analysed the intracellular delivery capacity of insect derived pyrrhocoricin with a peptide cargo in Cryptosporidium parvum in vitro using fluorescence microscopy. Results revealed that pyrrhocoricin was capable of acting as a delivery vehicle in transducing peptides across the parasite cell membrane for multiple life-cycle stages. The successful transduction may aid in target validation and the delivery of future peptide-based drugs against this important human pathogen.
Publisher: Elsevier BV
Date: 12-2015
Publisher: Public Library of Science (PLoS)
Date: 18-10-2013
Publisher: Elsevier BV
Date: 10-2014
DOI: 10.1016/J.EXPPARA.2014.07.008
Abstract: A new Caryospora coccidian species is described from the laughing kookaburra (Dacelo novaeguineae). Sporulated oocysts (n=30) are ovoid in shape with a smooth, colourless, bilayered oocyst wall and measure 31.4×29.3 (30.0-32.0×28.0-31.0) μm with a shape index of 1.1. Oocysts contain one spheroidal to subspheroidal sporocyst, 21.2×20.6 (20.0-24.0×20.0-21.0) μm. A spheroidal shaped sporocyst residuum is present micropyle, Stieda, substieda and parastieda bodies are absent. Vermiform sporozoites (n=8) are arranged either parallel or randomly in the sporocyst, measuring 17.0×4.8 (16.0-18.0×4.0-6.0) μm, with a L/W ratio of 3.5. There is a large spheroidal, posterior refractile body in the middle of the sporozoite. Morphologically, this new species is most similar to Caryospora. The prevalence of this parasite was 6.7% in birds s led in the morning and 33.3% from those s led after midday. Further molecular characterisation was conducted at two loci the 18S and 28S ribosomal RNA (rRNA). At the 18S locus, the new species of Caryospora was most closely related to Besnoitia besnoiti (99.2% similarity) and Hammondia triffittae (98.8% similarity). Although, no 28S partial sequences from Caryospora were available in GenBank, the highest similarity was with B.besnoiti (91.3%). Based on morphological and molecular data, this coccidian parasite is a new species that to date has not been reported. The new coccidian parasite is named Caryospora daceloe n. sp. after its host D. novaeguineae (the laughing kookaburra).
Publisher: Elsevier BV
Date: 06-2011
DOI: 10.1016/J.VETPAR.2011.02.011
Abstract: A total of 763 faecal s les were collected from western grey kangaroos (Macropus fuliginosus) in Western Australia and screened for the presence of Cryptosporidium by PCR at the 18S ribosomal RNA (rRNA) locus. S les that were positive at the 18S locus were also lified at the actin locus. The overall prevalence was 9.3% (71/763). At the 18S rRNA locus, sequences were obtained for 28 of the 71 positives. Sequence analysis identified four species Cryptosporidium fayeri in seven isolates, Cryptosporidium marcopodum in four isolates, Cryptosporidium xiaoi in six isolates and a novel genotype (kangaroo genotype I) in eleven isolates. Analysis at the actin locus confirmed the genetic distinctness of the novel genotype. The results of the present study indicate that in addition to C. fayeri and C. marcopodum, kangaroos may be capable of being infected with a wider range of Cryptosporidium species and genotypes including livestock species such as C. xiaoi. The novel genotype identified in the kangaroos most likely represents a cryptic species that requires further analyses to confirm its species status.
Publisher: Cambridge University Press (CUP)
Date: 07-2009
Abstract: Invasive aquatic weeds negatively affect bio ersity, fluvial dynamics, water quality, and water storage and conveyance for a variety of human resource demands. In California's Sacramento–San Joaquin River Delta, one submersed species—Brazilian egeria—and one floating species—waterhyacinth—are actively managed to maintain navigable waterways. We monitored the spatial and temporal dynamics of these species and their communities in the Sacramento-San Joaquin River Delta using airborne hyperspectral data and assessed the effect of herbicide treatments used to manage these species from 2003 to 2007. Each year, submersed aquatic plant species occupied about 12% of the surface area of the Delta in early summer and floating invasive plant species occupied 2 to 3%. Since 2003, the coverage of submersed aquatic plants expanded about 500 ha, whereas the coverage of waterhyacinth was reduced. Although local treatments have reduced the coverage of submersed aquatic plants, Delta-wide cover has not been significantly reduced. Locally, multiyear treatments could decrease submersed aquatic plants spread, given that no residual plants outside the treated area were present. In contrast, the spread of waterhyacinth either has been constant or has decreased over time. These results show that (1) the objectives of the Egeria densa Control Program (EDCP) have been hindered until 2007 by restrictions imposed on the timing of herbicide applications (2) submersed aquatic plants appeared to function as ecosystem engineers by enabling spread to adjacent areas typically subject to scouring action (3) repeated herbicide treatment of waterhyacinth has resulted in control of the spread of this species, which also appears to have facilitated the spread of waterprimrose and floating pennywort. These results suggest that management of the Delta aquatic macrophytes may benefit by an ecosystem-level implementation of an Integrated Delta Vegetation Management and Monitoring Program, rather than targeting only two problematic species.
Publisher: Elsevier BV
Date: 04-2016
Publisher: Wiley
Date: 12-2003
DOI: 10.1111/J.1751-0813.2003.TB14602.X
Abstract: Cryptosporidiosis is an enteric disease of animals and humans that can be fatal in immunocompromised in iduals. There is no known effective treatment for cryptosporidiosis. Bilbies are threatened marsupials and are bred in captivity as part of a recovery program to re-introduce this species to the southwest of Western Australia. Cryptosporidium muris infection was detected in the faeces of bilbies at a captive breeding colony. Stress associated with a high density of bilbies in enclosures may have predisposed some of the bilbies to infection with C. muris. C. muris has been described in mice and was found in the faeces of one mouse trapped in the breeding enclosures. It is likely the bilbies acquired the infection from mice by faecal contamination of food and water. The infection cleared within 2 months from some bilbies, however others remained infected for 6 months and treatment was attempted with dimetridazole. Subsequently the parasite was no longer be detectable in the faeces.
Publisher: Springer Science and Business Media LLC
Date: 10-09-2014
DOI: 10.1007/S00436-014-4118-Z
Abstract: Blood and ectoparasitic ticks were collected from migratory seabirds in New Zealand, including Australasian gannets (n = 13) from two sites and red-billed gulls (n = 9) and white-fronted terns (n = 2) from a third location. Blood smears were screened for parasite presence by microscopy, while DNA from blood s les was subjected to PCR for the presence of tick-transmitted protozoan haemoparasites belonging to the order Piroplasmida. Parasites were identified by comparing small subunit ribosomal RNA (18S rDNA) gene sequences to related sequences on GenBank. Analyses indicated that nine birds were infected with unknown variants of a Babesia poelea-like parasite (recorded as genotypes I and II), while four harboured a piroplasm that was genetically similar to Babesia kiwiensis. There was no parasite stratification by bird species both the gannets and gulls were positive for all three parasites, while the terns were positive for the B. kiwiensis-like and the B. poelea-like (genotype I) parasites. The B. kiwiensis-like parasite found in the birds was also found in two species of ticks: Carios capensis and Ixodes eudyptidis. This represents the first report of Babesia-positive ticks parasitising seabirds in New Zealand. The lack of host specificity and evidence of wide ranging distributions of the three piroplasm genotypes suggests there is a high degree of haemoparasite transmission occurring naturally between New Zealand seabird populations and species.
Publisher: American Society for Microbiology
Date: 06-2004
DOI: 10.1128/AEM.70.6.3761-3765.2004
Abstract: Histological, morphological, genetic, and phylogenetic analyses of a Cryptosporidium molnari -like isolate from a guppy ( Poecilia reticulata ) identified stages consistent with those of C. molnari and revealed that C. molnari is genetically very distinct from all other species of Cryptosporidium . This study represents the first genetic characterization of C. molnari .
Publisher: Elsevier BV
Date: 10-2020
Publisher: Elsevier BV
Date: 10-2012
DOI: 10.1016/J.EXPPARA.2012.07.003
Abstract: A total of 597 faecal s les were collected from western grey kangaroos (Macropus fuliginosus), Euros (M. robustus), red kangaroos (M. rufus) in Western Australia and Eastern Grey Kangaroos (M. giganteus) from Victoria and screened for the presence of Eimeria by PCR at the 18S ribosomal RNA (rRNA) locus. The overall prevalence was 24.3% (145/597). At the 18S rRNA locus, sequences were obtained for 25 of the 145 positives. Phylogenetic analysis indicated that all the macropod-derived Eimeria species grouped in a separate marsupial clade that included Eimeria trichosuri from brushtail possums. At least 6 different clades were identified within the marsupial isolates and many of the genotypes identified are likely to be valid species, however morphological and biological data need to be collected to match sequences to previously characterized Eimeria species or identify if they are new species.
Publisher: Cambridge University Press (CUP)
Date: 03-2002
DOI: 10.1079/JOH200198
Abstract: Confusion exists over the species status and host-specificity of the tapeworm Rodentolepis (= Hymenolepis ) nana . It has been described as one species, R. nana , found in both humans and rodents. Others have identified a subspecies R. nana var. fraterna , describing it as morphologically identical to the human form but only found in rodents. The species present in Australian communities has never been identified with certainty. Fifty one human isolates of Rodentolepis (= Hymenolepis ) nana were orally inoculated into Swiss Q, BALB/c, A/J, CBA/CAH and nude (hypothymic) BALB/c mice, Fischer 344 and Wistar rats and specific pathogen free (SPF) hamsters. Twenty four human isolates of R. nana were cross-tested in flour beetles, Tribolium confusum . No adult worms were obtained from mice, rats or hamsters, even when immunosuppressed with cortisone acetate. Only one of the 24 s les developed to the cysticercoid stage in T. confusum however, when inoculated into laboratory mice the cysticercoids failed to develop into adult worms. The large s le size used in this study, and the range of techniques employed for extraction and preparation of eggs provide a comprehensive test of the hypothesis that the human strain of R. nana is essentially non-infective to rodents.
Publisher: Elsevier BV
Date: 11-2014
DOI: 10.1016/J.TVJL.2014.08.001
Abstract: Faecal excretion of C ylobacter spp. and Salmonella enterica in sheep in Australia was determined using a quantitative multiplex PCR (qPCR) targeting the C ylobacter spp. purine biosynthesis gene (PurA) and the S. enterica outer membrane protein (ompF). The mutiplex qPCR was specific and C ylobacter spp. and S. enterica were each detected with a sensitivity of 5 organisms/µL faecal DNA extract. This multiplex qPCR was used to determine the prevalence and concentration of C ylobacter spp. and S. enterica in 3412 faecal s les collected from 1189 lambs on eight farms across South Australia (n = 2 farms), New South Wales (n = 1), Victoria (n = 2) and Western Australia (n = 3) at three s ling periods (weaning, post-weaning and pre-slaughter). The overall prevalences of C ylobacter spp. and S. enterica were 13.3% and 5.0%, respectively, with the highest prevalence for C ylobacter spp. in South Australia and the highest prevalence for S. enterica in New South Wales. C ylobacter jejuni was the only C ylobacter sp. identified from a subset of 120 positive s les sequenced at the 16S locus. S. enterica serovar Typhimurium was the only serovar of S. enterica identified from a subset of 120 positive s les sequenced at the ompF locus. Across all states, C ylobacter spp. had the highest median bacterial concentration in faeces at weaning and post-weaning (medians of 3.4 × 10(6) and 1.1 × 10(5), respectively), whereas S. enterica had the highest median bacterial concentration at pre-slaughter (1.8 × 10(5)/g faeces).
Publisher: Palacky University Olomouc
Date: 14-10-2016
DOI: 10.5507/FOT.2015.030
Publisher: Elsevier BV
Date: 06-2011
DOI: 10.1016/J.EXPPARA.2011.02.013
Abstract: To identify the animal sources for Cryptosporidium and Giardia contamination, we genotyped Cryptosporidium and Giardia spp. in wildlife from Sydney's water catchments using sequence analysis at the 18S rRNA locus for Cryptosporidium and 18S rRNA and glutamate dehydrogenase (gdh) for Giardia. A total of 564 faecal s les from 16 different host species were analysed. Cryptosporidium was identified in 8.5% (48/564) s les from eight host species and Giardia was identified in 13.8% (78/564) from seven host species. Eight species/genotypes of Cryptosporidium were identified. Five G. duodenalis assemblages were detected including the zoonotic assemblages A and B.
Publisher: Elsevier BV
Date: 09-2014
DOI: 10.1016/J.TVJL.2014.05.037
Abstract: The prevalence and faecal shedding of Chlamydia spp. in sheep in Australia has not been well described. Two species-specific quantitative PCRs (qPCRs) targeting the chlamydial outer membrane protein cell surface antigen gene (ompA) were validated and used to determine the prevalence and faecal shedding of C. abortus and C. pecorum from faecal s les of lambs at three s ling times (weaning, post-weaning and pre-slaughter) from eight farms in South Australia, New South Wales, Victoria and Western Australia. A total of 3412 faecal s les were collected and screened from approximately 1189 lambs across the four states. C. abortus was not detected in any of the s les screened. The overall prevalence of C. pecorum was 1027/3412 (30.1%) and median bacterial concentrations at weaning, post-weaning and pre-slaughter were 1.8 × 10(7), 1.2 × 10(7) and 9.6 × 10(5)/g faeces, respectively. A subset of C. pecorum positive s les from each farm, (n = 48) was sequenced to confirm their identity. The present study demonstrates that C. pecorum is prevalent in Australian sheep, highlighting a need for further research on the impact of this bacterium on production.
Publisher: Wiley
Date: 11-12-2007
DOI: 10.1111/J.1550-7408.2007.00299.X
Abstract: The morphology and infectivity of the oocysts of a new species of Cryptosporidium from the faeces of the red kangaroo (Macropus rufus) are described. Oocysts are structurally indistinguishable from those of Cryptosporidium parvum. Oocysts of the new species are passed fully sporulated, lack sporocysts, and measure 4.5-5.1 microm (mean=4.9) x 3.8-5.0 microm (mean=4.3 microm) with a length to width ratio 1.02:1.18 (mean 1.14) (n=50). Oocysts were not infectious for neonate ARC Swiss mice. Multi-locus analysis of numerous unlinked loci demonstrated this species to be distinct (90.64%-97.88% similarity) from C. parvum. Based on biological and molecular data, this Cryptosporidium infecting marsupials is proposed to be a new species Cryptosporidium fayeri n. sp.
Publisher: Springer Science and Business Media LLC
Date: 14-06-2016
Publisher: Elsevier BV
Date: 03-2007
DOI: 10.1016/J.VETPAR.2006.10.005
Abstract: Little is known about the species of Cryptosporidium infecting cats. The limited number of genetic studies conducted to date, have all identified C. felis as the species of Cryptosporidium in cats. We report a morphological and genetic description of a natural C. muris infection in a cat. Oocysts were viable and were successfully transmitted to laboratory mice. Further studies are required to determine the range and prevalence of Cryptosporidium species infecting cats.
Publisher: CRC Press
Date: 06-04-2015
DOI: 10.1201/B18317
Publisher: MDPI AG
Date: 14-10-2019
DOI: 10.3390/MICROORGANISMS7100452
Abstract: Cryptosporidium parvum is a protozoan parasite that can cause moderate-to-severe diarrhea. Insulinase-like proteases (INS) are one of the largest protein families within the small proteome of the pathogen. However, their roles in C. parvum biology remain un-elucidated. In this study, a member of the protein family, INS-15 of C. parvum encoded by cgd3_4260, was cloned, expressed and characterized to understand its function. INS-15 and its domain I were expressed in Escherichia coli and polyclonal antibodies against the domain I and one specific polypeptide were prepared in rabbits. The role of INS-15 protein in the C. parvum invasion was preliminarily studied. Recombinant INS-15 protein and its domain I were successfully expressed in E. coli, together with various degraded products. The cgd3_4260 gene had a peak expression at 2 h of in vitro C. parvum culture, while the INS-15 protein was expressed in the mid-anterior region of sporozoites and the area of merozoites opposite to the nucleus. Anti-INS-15 domain I antibodies reduced the invasion of C. parvum sporozoites by over 40%. The anterior location of INS-15 in invasion stages and partial reduction of in vitro growth indicate that INS-15 plays some roles in the invasion or early development of C. parvum.
Publisher: Cambridge University Press (CUP)
Date: 10-2002
DOI: 10.1017/S0031182002002172
Abstract: Iran is an important endemic focus of cystic hydatid disease (CHD) where several species of intermediate host are commonly infected with Echinococcus granulosus. Isolates of E. granulosus were collected from humans and other animals from different geographical areas of Iran and characterized using both DNA (PCR-RFLP of ITS1) and morphological criteria (metacestode rostellar hook dimensions). The sheep and camel strains/genotypes were shown to occur in Iran. The sheep strain was shown to be the most common genotype of E. granulosus affecting sheep, cattle, goats and occasionally camels. The majority of camels were infected with the camel genotype as were 3 of 33 human cases. This is the first time that cases of CHD in humans have been identified in an area where a transmission cycle for the camel genotype exists. In addition, the camel genotype was found to cause infection in both sheep and cattle. Results also demonstrated that both sheep and camel strains can be readily differentiated on the basis of hook morphology alone.
Publisher: Wiley
Date: 11-2002
DOI: 10.1111/J.1550-7408.2002.TB00224.X
Abstract: The structure and infectivity of the oocysts of a new species of Cryptosporidium from the feces of humans are described. Oocysts are structurally indistinguishable from those of Cryptosporidium parvum. Oocysts of the new species are passed fully sporulated, lack sporocysts. and measure 4.4-5.4 microm (mean = 4.86) x 4.4-5.9 microm (mean = 5.2 microm) with a length to width ratio 1.0-1.09 (mean 1.07) (n = 100). Oocysts were not infectious for ARC Swiss mice, nude mice. Wistar rat pups, puppies, kittens or calves, but were infectious to neonatal gnotobiotic pigs. Pathogenicity studies in the gnotobiotic pig model revealed significant differences in parasite-associated lesion distribution (P = 0.005 to P = 0.02) and intensity of infection (P = 0.04) between C. parvum and this newly described species from humans. In vitro cultivation studies have also revealed growth differences between the two species. Multi-locus analysis of numerous unlinked loci, including a preliminary sequence scan of the entire genome demonstrated this species to be distinct from C. parvum and also demonstrated a lack of recombination, providing further support for its species status. Based on biological and molecular data, this Cryptosporidium infecting the intestine of humans is proposed to be a new species Cryptosporidium hominis n. sp.
Publisher: Public Library of Science (PLoS)
Date: 18-12-2019
Publisher: Elsevier BV
Date: 06-1998
DOI: 10.1016/S0304-4017(98)00122-8
Abstract: Faecal s les were collected from domestic cats in the metropolitan area of the city of Perth, Western Australia, and screened for the presence of Cryptosporidium by both microscopy and PCR. Of 162 s les screened, two were positive for Cryptosporidium (a prevalence of 1.2%). S le Ct33 was from an 18-month-old female and s le Ct131 from a 12-month-old female. Morphological studies revealed oocysts with an average size of 4.6 x 4.0 microm, smaller in size than isolates typically seen in humans (5.0 x 4.5 microm). Sequence analysis of PCR products showed sequences from cat isolates to be different to previously sequenced human and calf isolates, with cat isolates exhibiting 8.1% sequence ergence from these isolates. Phylogenetic analysis grouped the cat isolates into a distinct group, separate from other C. parvum isolates and Cryptosporidium species. These results lend support to the existence of a cat-adapted Cryptosporidium strain or species.
Publisher: JSTOR
Date: 10-1996
DOI: 10.2307/3283880
Publisher: Elsevier BV
Date: 08-2018
DOI: 10.1016/J.VETPAR.2018.07.005
Abstract: Cryptosporidium and Giardia are common parasites of ruminant livestock worldwide. These parasites are associated with diarrhoea outbreaks in young goats (pre-weaning), but the impacts on health and productivity for older goats (post-weaning) are not well understood. Here we show Cryptosporidium faecal shedding is associated with reduced growth and diarrhoea in goats aged approximately 9-15 months. Goats were s led four times at one-month intervals. Faecal shedding for a range of pathogens were determined using quantitative PCR and sequencing (Cryptosporidium, Giardia, Eimeria, Salmonella, C ylobacter), and microscopy (trichostrongylid nematode worm egg count and Entamoeba). Cryptosporidium faecal shedding was associated with 1.5 kg lower growth for the one-month period following s ling. Specifically, C. xiaoi was associated with 1.9 kg lower growth in the following month. This is the first report of production impacts associated with C. xiaoi in ruminants older than 3 months of age. Cryptosporidium shedding was associated with an over 4-fold increase in risk of diarrhoea, with C. parvum associated with 10-fold and C. ubiquitum associated with 16-fold increase in risk of diarrhoea. Notably, C. xiaoi shedding was not associated with increased risk of diarrhoea. Giardia shedding was associated with looser faecal consistency, but not diarrhoea. Higher Eimeria oocyst counts were weakly associated with lower live weight, poorer body condition and looser faecal consistency. Shedding of other enteric pathogens were not associated with impacts on live weight, growth or diarrhoea risk. This study challenges the two notions that Cryptosporidium infections only impact health and productivity of goats during the pre-weaning period, and that Cryptosporidium (and specifically C. xiaoi) infections in the absence of diarrhoea are asymptomatic. Recognising the potential for impacts of Cryptosporidium infection on growth rates in the absence of diarrhoea will support improved design for experiments testing impacts of Cryptosporidium on ruminant health and production. Improved understanding of the role of protozoan infections on animal health has implications for the management of goats in order to reduce adverse impacts on farm profitability, animal welfare and public health risk.
Publisher: Elsevier BV
Date: 08-2012
DOI: 10.1016/J.EXPPARA.2012.04.015
Abstract: Cryptosporidiosis, caused by the protozoan parasite Cryptosporidium, represents the major public health concern of water utilities in developed nations due to its small size, resistance to disinfection and ability to be shed in large numbers in faeces. In Australia, recreational access is not allowed on direct supply sources, however, in Western Australia, limited recreational access to drinking water catchments has been allowed, although only in the outer catchment. Recreational activities within 2 km of the drinking water body is prohibited. The present study compared the amount, prevalence and species of Cryptosporidium in recreational versus non-recreational water catchments in the south west of Western Australia (WA). Recreational water catchments, which allowed swimming and c ing had a higher prevalence of Cryptosporidium and the majority of s les were the human-associated C. hominis. Non-recreational catchments had a lower prevalence and all the s les genotyped were C. parvum. Risk analysis identified increasing population as strongly correlated with an increase in the prevalence of Cryptosporidium in recreational catchments. This suggests that recreational access to drinking water catchments is a serious public health risk and government policy limiting activities to the outer catchment should be supported.
Publisher: Wiley
Date: 08-2007
Abstract: The accurate identification of Cryptosporidium (Protozoa: Apicomplexa) species and genotypes is central to the understanding of the transmission and to the diagnosis and control of cryptosporidiosis. In this study, we demonstrate the effectiveness of nonisotopic SSCP analysis of a approximately 300 bp region of the small subunit (pSSU) of ribosomal DNA for the specific identification of and delineation among 18 different Cryptosporidium species and genotypes from a wide range of hosts. This mutation scanning approach allowed the rapid and reliable differentiation between species/genotypes differing by as little as 1.3% in the pSSU sequence, with the capacity to detect point mutations. The present findings confirm the usefulness of this tool for the rapid genetic screening of Cryptosporidium s les from any host species, providing a foundation for detailed systematic, epidemiological and ecological studies. Although applied herein to pSSU, this low cost approach should be applicable to a wide range of genetic loci for population genetic investigations of Cryptosporidium.
Publisher: Microbiology Society
Date: 10-2016
Abstract: Recently, two novel species of Anaplasmataceae were detected in the Australian paralysis tick, Ixodes holocyclus, by 16S rRNA gene metabarcoding. Analysis of these sequences suggested that these novel organisms are closely related to the genus 'Candidatus Neoehrlichia'. In this study, phylogenetic analysis of 16S rRNA (1264 bp), groESL (1047 bp) and gltA (561 bp) gene sequences, and concatenated (2872 bp) sequences, all concur that these novel species belong in the genus 'Candidatus Neoehrlichia' and are most closely related to, but distinct from the only other recognised members of this genus, 'Candidatus Neoehrlichia mikurensis' and 'Candidatus Neoehrlichia lotoris'. Based on their unique molecular signature, we propose to designate these species 'Candidatus Neoehrlichia australis' (reference strain HT41R) and 'Candidatus Neoehrlichia arcana' (reference strain HT94R). Identical 'Candidatus Neoehrlichia australis' 16S rRNA, groESL and gltA sequences were detected in 34/391 (8.7 %) in idual Ixodes holocyclus ticks, and sequences were most similar to 'Candidatus Neoehrlichia lotoris' (96.2 %, 83.1 % and 67.2 %, respectively) and 'Candidatus Neoehrlichia mikurensis' (96.2 %, 84 % and 68.4 % respectively). Likewise, identical 'Candidatus Neoehrlichia arcana' 16S rRNA, groESL and gltA sequences were detected in 12/391 (3.1 %) Ixodes holocyclus ticks, and sequences were most similar to 'Candidatus Neoehrlichia lotoris' (98.5 %, 88.7 % and 79.3 %, respectively) and 'Candidatus Neoehrlichia mikurensis' (96.3 %, 84 % and 67.4 % respectively). These new species are the first Anaplasmataceae (except Wolbachia spp.) to be found to be endemic to Australia. The pathogenic consequences of these organisms are yet to be determined.
Publisher: Elsevier BV
Date: 03-2013
DOI: 10.1016/J.VETPAR.2012.12.008
Abstract: The third-stage larvae of several genera of anisakid nematodes are important etiological agents for zoonotic human anisakiasis. The present study investigated the prevalence of potentially zoonotic anisakid larvae in fish collected on the coastal shelves off Madang and Rabaul in Papua New Guinea (PNG) where fish represents a major component of the diet. Nematodes were found in seven fish species including Decapterus macarellus, Gerres oblongus, Pinjalo lewisi, Pinjalo pinjalo, Selar crumenophthalmus, Scomberomorus maculatus and Thunnus albacares. They were identified by both light and scanning electron microscopy as Anisakis Type I larvae. Sequencing and phylogenetic analysis of the ribosomal internal transcribed spacer (ITS) and the mitochondrial cytochrome C oxidase subunit II (cox2) gene identified all nematodes as Anisakis typica. This study represents the first in-depth characterisation of Anisakis larvae from seven new fish hosts in PNG. The overall prevalence of larvae was low (7.6%) and no recognised zoonotic Anisakis species were identified, suggesting a very low threat of anisakiasis in PNG.
Publisher: Elsevier BV
Date: 12-1997
Publisher: IWA Publishing
Date: 02-2014
Publisher: IWA Publishing
Date: 10-2022
DOI: 10.2166/WH.2022.114
Abstract: Contamination of drinking water from Norovirus (NoV) and other waterborne viruses is a major public health concern globally. Increasingly, quantitative microbial risk assessment (QMRA) is being used to assess the various risks from waterborne pathogens and evaluate control strategies. As urban populations grow and expand, there is increasing demand for recreational activities in drinking water catchments. QMRA relies on context-specific data to map out the pathways by which viruses can enter water and be transferred to drinking water consumers and identify risk factors and appropriate controls. This review examines the current evidence base and assumptions for QMRA analysis of NoV and other waterborne viral pathogens and recommends numerical values based on the most recent evidence to better understand the health risks associated with recreators in Australian drinking water sources these are broadly applicable to all drinking water sources where recreational access is allowed. Key issues include the lack of an agreed upon data and dose-response models for human infectious NoV genotypes, faecal shedding by bathers, the extent of NoV infectivity and aggregation, resistance (secretor status) to NoV and the extent of secondary transmission.
Publisher: Elsevier BV
Date: 05-2015
DOI: 10.1016/J.IJPARA.2015.01.009
Abstract: Cryptosporidium is an enteric parasite that is considered the second greatest cause of diarrhoea and death in children after rotavirus. Currently, 27 species are recognised as valid and of these, Cryptosporidium hominis and Cryptosporidium parvum are responsible for the majority of infections in humans. Molecular and biological studies indicate that Cryptosporidium is more closely related to gregarine parasites rather than to coccidians. The identification of gregarine-like gamont stages and the ability of Cryptosporidium to complete its life cycle in the absence of host cells further confirm its relationship with gregarines. This opens new avenues into the investigation of pathogenesis, epidemiology, treatment and control of Cryptosporidium. Effective drug treatments and vaccines are not yet available due, in part, to the technical challenges of working on Cryptosporidium in the laboratory. Whole genome sequencing and metabolomics have expanded our understanding of the biochemical requirements of this organism and have identified new drug targets. To effectively combat this important pathogen, increased funding is essential.
Publisher: Elsevier BV
Date: 07-2016
DOI: 10.1016/J.EXPPARA.2016.04.011
Abstract: A new Isospora (Apicomplexa: Eimeriidae) species is described from a single red-browed finch (Neochmia temporalis) (subspecies N. temporalis temporalis), that was part of a captive population in Western Australia. Sporulated oocysts of this isolate are spherical, 18.3 (18.2-18.9) × 18.2 (18.2-18.6) μm, with a shape index (length/width) of 1.0 and a smooth and bilayered oocyst wall, 1.2 μm thick (outer layer 0.9 μm, inner 0.3 μm). A polar granule is present, but the oocyst residuum and a micropyle are absent. The sporocysts are ovoid-shaped, 13.3 (9.5-16.4) × 8.6 (6.8-10.0) μm, with a shape index of 1.5. An indistinct Stieda body is present, but the substieda body is absent. A sporocyst residuum is present and composed of numerous granules of different size scattered among the sporozoites. Morphologically, the oocysts from this isolate are different from those of all known valid Isospora spp. Molecular analysis was conducted at 4 loci the 18S and 28S ribosomal RNA (rRNA), the mitochondrial cytochrome oxidase (COI) gene and the heat shock protein 70 (hsp70) gene. At the 18S locus, this new isolate exhibited 99.9%, 99.8%, 99.7%, and 99.5% similarity to I. sp. MAH-2013a from a superb starling (L rotornis superbus), I. MS-2003 from a Southern cape sparrow (Passer melanurus), I. sp. Tokyo from a domestic pigeon (Columba livia domestica) and I. MS-2003 from a Surinam crested oropendula (Psarocolius decumanus). At the 28S locus, this new isolate exhibited 99.7% similarity to both an Isospora sp (MS-2003) from a Northern house sparrow (Passer domesticus) and an Isospora sp. (MS-2003) from a Southern cape sparrow. At the COI locus, this new isolate exhibited 98.9% similarity to an Isospora sp. ex Apodemus flavicollis. At the hsp70 locus, this new isolate exhibited 99% similarity to isolate MS-2003 (AY283879) from a wattled starling (Creatophora cinerea). Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora neochmiae n. sp. after its host, the red-browed finch (Neochmia temporalis).
Publisher: Elsevier BV
Date: 08-2019
Publisher: Elsevier BV
Date: 04-2015
DOI: 10.1016/J.EXPPARA.2015.02.001
Abstract: Cryptosporidium is an important enteric pathogen that infects a wide range of humans and animals. Rapid and reliable detection and characterisation methods are essential for understanding the transmission dynamics of the parasite. Sanger sequencing, and high-throughput sequencing (HTS) on an Ion Torrent platform, were compared with each other for their sensitivity and accuracy in detecting and characterising 25 Cryptosporidium-positive human and animal faecal s les. Ion Torrent reads (n = 123,857) were obtained at both 18S rRNA and actin loci for 21 of the 25 s les. Of these, one isolate at the actin locus (Cattle 05) and three at the 18S rRNA locus (HTS 10, HTS 11 and HTS 12), suffered PCR drop-out (i.e. PCR failures) when using fusion-tagged PCR. Sanger sequences were obtained for both loci for 23 of the 25 s les and showed good agreement with Ion Torrent-based genotyping. Two s les both from pythons (SK 02 and SK 05) produced mixed 18S and actin chromatograms by Sanger sequencing but were clearly identified by Ion Torrent sequencing as C. muris. One isolate (SK 03) was typed as C. muris by Sanger sequencing but was identified as a mixed C. muris and C. tyzzeri infection by HTS. 18S rRNA Type B sequences were identified in 4/6 C. parvum isolates when deep sequenced but were undetected in Sanger sequencing. Sanger was cheaper than Ion Torrent when sequencing a small numbers of s les, but when larger numbers of s les are considered (n = 60), the costs were comparative. Fusion-tagged licon based approaches are a powerful way of approaching mixtures, the only draw-back being the loss of PCR efficiency on low-template s les when using primers coupled to MID tags and adaptors. Taken together these data show that HTS has excellent potential for revealing the "true" composition of species/types in a Cryptosporidium infection, but that HTS workflows need to be carefully developed to ensure sensitivity, accuracy and contamination are controlled.
Publisher: No publisher found
Date: 2005
Publisher: Elsevier BV
Date: 12-2001
Publisher: Springer Science and Business Media LLC
Date: 21-05-2016
Publisher: Elsevier BV
Date: 03-2016
Publisher: Elsevier BV
Date: 03-2016
Publisher: Elsevier BV
Date: 2018
Publisher: Elsevier BV
Date: 12-2018
DOI: 10.1016/J.SCITOTENV.2018.07.024
Abstract: Wastewater recycling is an increasingly popular option in worldwide to reduce pressure on water supplies due to population growth and climate change. Cryptosporidium spp. are among the most common parasites found in wastewater and understanding the prevalence of human-infectious species is essential for accurate quantitative microbial risk assessment (QMRA) and cost-effective management of wastewater. The present study conducted next generation sequencing (NGS) to determine the prevalence and ersity of Cryptosporidium species in 730 raw influent s les from 25 Australian wastewater treatment plants (WWTPs) across three states: New South Wales (NSW), Queensland (QLD) and Western Australia (WA), between 2014 and 2015. All s les were screened for the presence of Cryptosporidium at the 18S rRNA (18S) locus using quantitative PCR (qPCR), oocyst numbers were determined directly from the qPCR data using DNA standards calibrated by droplet digital PCR, and positives were characterized using NGS of 18S licons. Positives were also screened using C. parvum and C. hominis specific qPCRs. The overall Cryptosporidium prevalence was 11.4% (83/730): 14.3% (3/21) in NSW 10.8% (51/470) in QLD and 12.1% (29/239) in WA. A total of 17 Cryptosporidium species and six genotypes were detected by NGS. In NSW, C. hominis and Cryptosporidium rat genotype III were the most prevalent species (9.5% each). In QLD, C. galli, C. muris and C. parvum were the three most prevalent species (7.7%, 5.7%, and 4.5%, respectively), while in WA, C. meleagridis was the most prevalent species (6.3%). The oocyst load/Litre ranged from 70 to 18,055 oocysts/L (overall mean of 3426 oocysts/L: 4746 oocysts/L in NSW 3578 oocysts/L in QLD and 3292 oocysts/L in WA). NGS-based profiling demonstrated that Cryptosporidium is prevalent in the raw influent across Australia and revealed a large ersity of Cryptosporidium species and genotypes, which indicates the potential contribution of livestock, wildlife and birds to wastewater contamination.
Publisher: Elsevier BV
Date: 2015
DOI: 10.1016/J.EXPPARA.2014.11.003
Abstract: Cryptosporidiosis, a gastroenteric disease characterised mainly by diarrheal illnesses in humans and mammals is caused by infection with the protozoan parasite Cryptosporidium. Treatment options for cryptosporidiosis are limited, with the current therapeutic nitazoxanide, only partly efficacious in immunocompetent in iduals. The parasite lacks de novo purine synthesis, and is exclusively dependant on purine salvage from its host. Inhibition of the inosine 5' monophosphate dehydrogenase (IMPDH), a purine salvage enzyme that is essential for DNA synthesis, thereby offers a potential drug target against this parasite. In the present study, a yeast-two-hybrid system was used to identify Phylomer peptides within a library constructed from the genomes of 25 phylogenetically erse bacteria that targeted the IMPDH of Cryptosporidium parvum (IMPcp) and Cryptosporidium hominis (IMPch). We identified 38 unique interacting Phylomers, of which, 12 were synthesised and screened against C. parvum in vitro. Two Phylomers exhibited significant growth inhibition (81.2-83.8% inhibition P < 0.05), one of which consistently exhibited positive interactions with IMPcp and IMPch during primary and recapitulation yeast two-hybrid screening and did not interact with either of the human IMPDH proteins. The present study highlightsthe potential of Phylomer peptides as target validation tools for Cryptosporidium and other organisms and diseases because of their ability to bind with high affinity to target proteins and disrupt function.
Publisher: American Society for Microbiology
Date: 02-2004
DOI: 10.1128/AEM.70.2.891-899.2004
Abstract: The genetic ersity of Cryptosporidium in reptiles was analyzed by PCR-restriction fragment length polymorphism and sequence analysis of the small subunit rRNA gene. A total of 123 s les were analyzed, of which 48 snake s les, 24 lizard s les, and 3 tortoise s les were positive for Cryptosporidium . Nine different types of Cryptosporidium were found, including Cryptosporidium serpentis , Cryptosporidium desert monitor genotype, Cryptosporidium muris , Cryptosporidium parvum bovine and mouse genotypes, one C. serpentis -like parasite in a lizard, two new Cryptosporidium spp. in snakes, and one new Cryptosporidium sp. in tortoises. C. serpentis and the desert monitor genotype were the most common parasites and were found in both snakes and lizards, whereas the C. muris and C. parvum parasites detected were probably the result of ingestion of infected rodents. Sequence and biologic characterizations indicated that the desert monitor genotype was Cryptosporidium saurophilum . Two host-adapted C. serpentis genotypes were found in snakes and lizards.
Publisher: Frontiers Media SA
Date: 15-05-2019
Publisher: Elsevier BV
Date: 10-2012
DOI: 10.1016/J.MOLBIOPARA.2012.08.006
Abstract: Faecal metabolite profiling, though in its infancy, allows for investigation of complex metabolic interactions between gastrointestinal infections or diseases and host health. In the present study, we describe a faecal metabolite extraction method for untargeted gas chromatography-mass spectrometry (GC-MS) analysis using Cryptosporidium positive and negative human faecal s les. The extraction method takes into account the varying faecal consistencies and quantities received for clinical diagnosis. Optimisation was carried out using different extraction solvents and on three different faecal quantities to determine the minimum amount of faecal s le required. The method was validated by untargeted GC-MS analysis on 8 Cryptosporidium positive and 8 Cryptosporidium negative human faecal s les, extracted using the optimised conditions. The method showed good extraction reproducibility with a relative standard deviation of 9.14%. Multivariate analysis of the GC-MS generated dataset showed distinct differences between profiles of Cryptosporidium positive and Cryptosporidium negative s les. The most notable differences included changes in amino acid, nitrogen and energy metabolism, demonstrating the association of infection with Cryptosporidium and altered permeability of the small intestine.
Publisher: Elsevier BV
Date: 11-2017
DOI: 10.1016/J.MEEGID.2017.09.025
Abstract: Cryptosporidium and Giardia are major causes of diarrhoea in developing countries including Ghana, however, nothing is known about the species and subtypes of Cryptosporidium and Giardia in farmers and their ruminant livestock in this country. A total of 925 faecal s les from humans (n=95), cattle (n=328), sheep (n=217) and goats (n=285), were screened for Cryptosporidium and Giardia by quantitative PCR (qPCR) at the 18S rRNA and glutamate dehydrogenase (gdh) loci respectively. Cryptosporidium positives were typed by sequence analysis of 18S and 60kDa glycoprotein (gp60) loci licons. Giardia positives were typed at the triose phosphate isomerase (tpi), beta-giardin (bg) and gdh loci. The prevalence of Cryptosporidium and Giardia by qPCR was 8.4% and 10.5% in humans, 26.5% and 8.5% in cattle, 34.1% and 12.9% in sheep, and 33.3% and 12.3% in goat faecal s les, respectively. G. duodenalis assemblages A and B were detected in humans and assemblage E was detected in livestock. Cryptosporidium parvum was the only species identified in humans C. andersoni, C. bovis, C. ryanae and C. ubiquitum were identified in cattle C. xiaoi, C. ubiquitum and C. bovis in sheep and C. xiaoi, C. baileyi and C. parvum in goats. This is the first molecular study of Cryptosporidium and Giardia in livestock in Ghana. The identification of zoonotic species and the identification of C. parvum subtype IIcA5G3q in livestock, which has previously been identified in children in Ghana, suggests potential zoonotic transmission. Further studies on larger numbers of human and animal s les, and on younger livestock are required to better understand the epidemiology and transmission of Cryptosporidium and Giardia in Ghana.
Publisher: Elsevier BV
Date: 02-2018
DOI: 10.1016/J.TTBDIS.2017.12.011
Abstract: Worldwide, Ehrlichia spp. are emerging infectious organisms of domestic animals and people, however, most Ehrlichia spp. naturally infect wildlife reservoirs causing mainly asymptomatic infections. Australian ecosystems have been under-explored for these potentially pathogenic organisms, and recent studies have identified a range of novel Ehrlichia, and their sister genera, Anaplasma and 'Candidatus Neoehrlichia' species, from native Australian ticks. We used bacterial 16S rRNA (16S) next-generation sequencing and genus-specific PCR to profile the bacterial communities in platypus (Ornithorhynchus anatinus) blood s les and platypus ticks (Ixodes ornithorhynchi), and identified a high prevalence of Ehrlichia sequences. We also observed Ehrlichia-like intra-neutrophilic inclusions (morulae) in PCR-positive stained platypus blood films that were consistent in morphology with other Ehrlichia spp. Bayesian phylogenetic analysis of 16S (1343 bp), gltA (1004 bp), and groEL (1074 bp) gene sequences group the platypus Ehrlichia with 'Candidatus Ehrlichia khabarensis' from far-eastern Russia, and demonstrate that the platypus Ehrlichia is clearly distinct from all other Ehrlichia spp. Enough genetic ergence exists to delineate this platypus Ehrlichia as a separate species that we propose to designate 'Candidatus Ehrlichia ornithorhynchi'. There is no evidence that 'Candidatus Ehrlichia ornithorhynchi' causes disease in wild platypuses, however, the organism does seem to be widespread in Australia, being found in both Queensland and Tasmania. 'Candidatus Ehrlichia ornithorhynchi' is the second native Australian Ehrlichia described and adds to the rapidly growing ersity of recently described native Australian tick-borne bacteria.
Publisher: Public Library of Science (PLoS)
Date: 14-12-2016
Publisher: Springer Science and Business Media LLC
Date: 28-10-2021
DOI: 10.1007/S11686-021-00482-5
Abstract: There is a dearth of research conducted on the Knowledge, Attitude and Practices (KAP) of swimming pool patrons and staff to determine their understanding of the importance of Cryptosporidium and its transmission in swimming pools. We conducted a KAP survey of public swimming pool patrons (n = 380) and staff (n = 40) attending five public swimming pools in Western Australia (WA). Knowledge, attitudes and practices (KAP) of Cryptosporidium varied between patrons and staff but were generally limited. Only 26.1% and 25.0% of patrons and staff had heard of Cryptosporidium, while 17.4% and 10.0% knew that it causes diarrhoea, respectively. Thirty-one percent of patrons were aware of their pool policy concerning gastroenteritis and Cryptosporidium, compared to 62.5% of staff. Less than 50% of patrons demonstrated awareness of how features within the pool environment were relevant to the control of Cryptosporidium. Only about a third of patrons (35%) and staff (37.5%) were aware that showering before swimming reduced the risk of gastroenteritis. Raising awareness about hygiene-related practices through the delivery of targeted health education messages to the general public is essential to reduce the burden of Cryptosporidium infections in aquatic environments.
Publisher: Cambridge University Press (CUP)
Date: 1999
DOI: 10.1017/S0031182098003412
Abstract: The Cryptosporidium ITS1, 5·8S and ITS2 rDNA regions from a number of Cryptosporidium isolates from different hosts and geographical areas were cloned and sequenced in order to investigate the extent of sequence heterogeneity between human and cattle-derived isolates from different geographical locations and also between isolates of Cryptosporidium from different hosts such as cats, pigs, mice and a koala. Calf-derived isolates from different continents were virtually identical as were human-derived isolates from the UK and Australia. Genetic differences between Cryptosporidium isolates were extensive and were in fact greater than the level of nucleotide ergence between Toxoplasma gondii and Neospora caninum rDNA sequences. Based on the sequence information derived from this study, PCR–RFLP of the ITS1 region was undertaken in order to directly lify and genotype Cryptosporidium isolates from different hosts. This PCR–RFLP approach can now be used for molecular epidemiology studies, circumventing the need for costly sequencing and allowing a wider range of genetically different isolates to be examined.
Publisher: American Society for Microbiology
Date: 12-2007
DOI: 10.1128/AEM.00848-07
Abstract: A total of 250 mouse fecal specimens collected from crop farms in Queensland, Australia, were screened for the presence of Cryptosporidium spp. using PCR. Of these, 19 positives were detected and characterized at a number of loci, including the 18S rRNA gene, the acetyl coenzyme A gene, and the actin gene. Sequence and phylogenetic analyses identified two genotypes: mouse genotype I and a novel genotype (mouse genotype II), which is likely to be a valid species. Cryptosporidium parvum , which is zoonotic, was not detected. The results of the study indicate that wild Australian mice that are not in close contact with livestock are probably not an important reservoir of Cryptosporidium infection for humans and other animals.
Publisher: Elsevier BV
Date: 12-2002
DOI: 10.1016/S0020-7519(02)00199-6
Abstract: The present study describes the complete development of all life cycle stages of Cryptosporidium andersoni in the HCT-8 cell line. The in vitro cultivation protocols were the same as those used for the successful growth of all life cycle stages of Cryptosporidium parvum (Int. J. Parasitol. 31 (2001) 1048). Under these culture conditions, C. andersoni grew and proliferated rapidly with the completion of the entire life cycle within 72h post-infection. The developmental stages of C. andersoni are larger than those of C. parvum enabling easier identification of life cycle stages including a previously unrecognised extracellular stage. The presence of this extracellular stage was further confirmed following its isolation from the faeces of infected cattle using a laser microdissection technique. This stage was present in large numbers and some of them were seen undergoing syzgy. Extraction of DNA from the extracellular stage, followed by polymerase chain reaction-restriction fragment length polymorphism and sequencing of the 18S rDNA confirmed that this is a stage in the life cycle of C. andersoni. In vitro, extracellular stages were always observed moving over the HCT-8 cells infected with C. andersoni. Comparative observations with C. parvum also confirmed the presence of extracellular stages. Extracellular stages were recovered from in vitro culture after 5 days post-infection with the cattle genotype of C. parvum and from infected mice. At least two morphologically different stages (stages one and two) were purified from mice after 72h of infection. The presence and morphological characterisation of extracellular developmental stages in the life cycle of Cryptosporidium confirms its relationship to gregarines and provides important implications for our understanding of the taxonomic and phylogenetic affinities of the genus Cryptosporidium. The growth of C. andersoni in cell culture now provides a means of studying its development, metabolism, and behaviour as well as testing its response to different therapeutic agents.
Publisher: Elsevier BV
Date: 05-2020
Publisher: Elsevier BV
Date: 09-2016
DOI: 10.1016/J.VETPAR.2016.08.015
Abstract: Little is known about the epidemiology of Cryptosporidium in Jordan and to date, only one genotyping study has been conducted on Cryptosporidium isolates from Jordanian children. In the present study, a total of 284 faecal s les from Jordanian cattle, sheep, goats and chicken and 48 human faecal s les were screened for the presence of Cryptosporidium using an 18S quantitative PCR (qPCR) and a C. parvum/C. hominis specific qPCR at a lectin locus. Of these, 37 of 284 animal faecal s les were positive by qPCR at the 18S locus giving an overall prevalence of 11.6%. The point prevalence of Cryptosporidium in chickens, sheep, horses, cattle and goats ranged from 4.8% (chickens) to 18.7% (cattle). A total of six species were detected C. xiaoi (n=9),C. andersoni (n=7),C. ryanae (n=5),C. parvum (n=4),C. baileyi (n=1) and a genetically distinct and potentially novel species in two isolates from horses. Sub-genotype analysis of the 4 C. parvum isolates at the 60-kDa glycoprotein (gp60) locus identified subtype IIaA19G2R1 (n=2) and IIaA16GR1 (n=2). For the human s les, 4 positives (8.3% prevalence) were detected. Of these, two were C. parvum (subtypes IIdA20G1 and IIaA15G2R1) and two were C. hominis (subtypes 1bA9G3 and 1bA10G2R2). Further studies are required to better understand the epidemiology and transmission of Cryptosporidium in Jordan.
Publisher: Elsevier BV
Date: 09-2011
DOI: 10.1016/J.EXPPARA.2011.06.009
Abstract: Cryptosporidium oocysts were inoculated into fresh dung (∼1.2×10(4) oocysts per gram wet weight) and fed to dung beetles to assess the effect of dung burial by the dung beetle Bubas bison on the distribution of the oocysts in small cores of soil in the laboratory. The experiment consisted of five replicates of each of two treatments controls (dung but no dung beetles) and the experimental treatment (inoculated dung and seven pairs of dung beetles). After 5 days, when approximately 90% of the dung was buried, the surface and buried dung was recovered and subs led. The oocysts in the subs les were recovered and enumerated using qPCR. Oocyst viability was evaluated using an assay based on the exclusion or inclusion of two fluorogenic vital dyes, 4',6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI). Results revealed that overall 13.7% of oocysts remained on the surface and 86.3% of oocysts were buried. The viability of oocysts in buried dung was only 10% compared to oocysts the surface dung (58%). Therefore, widespread dung burial by B. bison during the winter months could substantially reduce the numbers of Cryptosporidium oocysts available to be washed into waterways following winter rains.
Publisher: Elsevier BV
Date: 11-2015
Publisher: Elsevier
Date: 2003
Publisher: Cambridge University Press (CUP)
Date: 05-2000
DOI: 10.1017/S0031182099005703
Abstract: Isolates of Cryptosporidium muris and C. serpentis were characterized from different hosts using nucleotide sequence analysis of the rDNA 18S and ITS1 regions, and the heat-shock (HSP-70) gene. Phylogenetic analysis confirmed preliminary evidence that C. muris is not a uniform species. Two distinct genotypes were identified within C. muris (1) C. muris genotype A comprising bovine and camel isolates of C. muris from different geographical locations, and (2) C. muris genotype B comprising C. muris isolates from mice, a hamster, a rock hyrax and a camel from the same enclosure. These 2 genotypes may represent separate species but further biological and molecular studies are required for confirmation.
Publisher: Centers for Disease Control and Prevention (CDC)
Date: 02-2014
Publisher: Springer Science and Business Media LLC
Date: 20-04-2017
Publisher: Elsevier BV
Date: 11-2000
DOI: 10.1016/S0020-7519(00)00129-6
Abstract: Understanding the epidemiology of zoonotic parasitic infections is dependent upon the availability of accurate and sensitive diagnostic techniques. The development of molecular diagnostic methods, particularly those utilising PCR for the detection of zoonoses will contribute greatly to the identification and control of these pathogens, by increasing the speed of diagnosis, specificity and sensitivity, reproducibility and ease of interpretation. Molecular characterisation studies allow us to distinguish between closely related infectious agents and to document the patterns of transmission of 'strains' and species within populations. This will allow precise determinations to be made about the aetiological agent, its characteristics and the source of infection. This review focuses on recent detection and characterisation techniques for both emerging and re-emerging parasite zoonoses.
Publisher: Springer Science and Business Media LLC
Date: 11-08-2020
Publisher: Elsevier BV
Date: 04-2009
DOI: 10.1016/J.EXPPARA.2009.01.008
Abstract: Little is known of the prevalence of Giardia species and genotypes in pre- and post-weaned domestic pigs. In the present study, a total of 297 pig faecal s les were screened for the presence of Giardia by PCR and genotyped. An overall prevalence of 31.1% (90/289) (25.8, 36.5 CI) was detected. Giardia was detected in 17% (23/123) (11.8-25.6 CI) of pre-weaned piglet faecal s les and 41% (64/156) (33.3-48.7 CI) post-weaned faecal s les analysed. Sequence analysis identified assemblage A and E in pre- and post-weaned pigs. Assemblage F was identified in one post-weaned pig. Assemblage E was the most prevalent assemblage detected.
Publisher: Elsevier BV
Date: 04-2010
DOI: 10.1016/J.VETPAR.2009.12.021
Abstract: The present study reports for the first time the completion of the life cycle of Cryptosporidium hominis in cell-free culture and multiplication of the parasite via qPCR. In idual life-cycle stages were characterised using Cryptosporidium-specific antibody staining (Sporo-Glo) and fluorescent in situ hybridisation (FISH) staining on cultures inoculated with excysted oocysts and purified sporozoites. In both cultures, C. hominis successfully proliferated and completed its life cycle, however development in cultures inoculated with purified sporozoites lagged behind cultures inoculated with excysted oocysts. Some novel findings of the study include the visualisation of pairing and multiple associations between various developmental stages in a process similar to syzygy and the formation of Cryptosporidium stages (trophozoites and meronts) inside the oocysts without excystation. qPCR analysis revealed a 5-6-fold lification of parasite DNA. Future studies are required to improve the lification of the parasite. The present study confirms the suitability of this culturing model to support the growth and proliferation of C. hominis (which unlike C. parvum, cannot be readily cultured in small animal models) and will greatly assist in our understanding of the developmental biology of Cryptosporidium, its position within the Apicomplexa and its relationship to gregarine protozoa.
Publisher: Elsevier BV
Date: 12-2015
DOI: 10.1016/J.VETPAR.2015.10.013
Abstract: Cryptosporidium is an enteric parasite that infects humans and a wide range of animals. Relatively little is known about the epidemiology and taxonomy of Cryptosporidium in fish. In the present study, a total of 775 fish, belonging to 46 species and comprising ornamental fish, marine fish and freshwater fish were screened for the prevalence of Cryptosporidium by PCR. The overall prevalence of Cryptosporidium in fish was 5.3% (41/775), with prevalences ranging from 1.5 to 100% within in idual host species. Phylogenetic analysis of these Cryptosporidium isolates as well as 14 isolates from previous studies indicated extensive genetic ersity as well as evidence for mixed infections. At the 18S locus the following species were identified Cryptosporidium molnari-like genotype (n=14), Cryptosporidium huwi (n=8), piscine genotype 2 (n=4), piscine genotype 3-like (n=1), piscine genotype 4 (n=2), piscine genotype 5 (n=13), piscine genotype 5-like (n=1) and five novel genotypes (n=5). At the actin locus, species identification agreed with the 18S locus for only 52.3% of isolates sequenced, indicating high levels of mixed infections. Future studies will need to employ both morphological characterization and deep sequencing licon-based technologies to better understand the epidemiological and phylogenetic relationships of piscine-derived Cryptosporidium species and genotypes, particularly when mixed infections are detected.
Publisher: Elsevier BV
Date: 12-2020
Publisher: Canadian Science Publishing
Date: 06-2002
DOI: 10.1139/W02-047
Abstract: The environmental distribution of Giardia spp. and Cryptosporidium spp. is dependent upon human, agricultural, and wildlife sources. The significance of each source with regard to the presence of parasites in the environment is unknown. This 2-year study examined parasite prevalence in human sewage influent, wildlife, and agricultural sources associated with the North Saskatchewan River Basin in Alberta, Canada. Fecal s les were collected from cowcalf, dairy, and hog operations in the watershed area. Sewage-treatment facilities were s led bimonthly during the 2-year study, and wildlife scat was collected at locations along tributaries of the North Saskatchewan River. All s les were analyzed for the presence of Giardia and Cryptosporidium, using sucrose-gradient separation followed by immunofluorescent microscopy. Giardia and Cryptosporidium were detected in all three sources. The lowest prevalence of both Giardia (3.28%) and Cryptosporidium (0.94%) was found in wildlife, with 6 of 19 species testing positive. Sewage influent had the highest prevalence of Giardia (48.80%) and Cryptosporidium parvum-like oocysts (5.42%) however, the concentration of both parasites was minimal compared with the concentration detected in cattle feces. Cowcalf sources contained the highest concentration of Giardia (mean 5800/g feces, P 0.01), and dairy sources contained the highest concentration of C. parvum-like oocysts (mean 295/g feces, P 0.01). Although prevalence and concentration are higher in cattle feces than in sewage, the Giardia and Cryptosporidium in animal manure do not have direct access to water draining into the North Saskatchewan River. PCR-based characterization of rDNA from isolates ofCryptosporidium collected from Alberta human, pig, calf, mature steer, dog, cat, and beaver hosts revealed distinct genetic differences that may reflect host specificity.Key words: Giardia , Cryptosporidium, cattle, pigs, PCR, diagnostics, waterborne parasites.
Publisher: Elsevier BV
Date: 03-2022
DOI: 10.1016/J.MEEGID.2022.105212
Abstract: Cryptosporidium spp. and Giardia duodenalis are important protozoan parasites which are associated with diarrheal diseases in humans and animals worldwide. Relatively little is known about the molecular epidemiology of Cryptosporidium spp. and Giardia duodenalis in the Middle East Countries and North Africa (MENA region). Therefore, this review aimed to inspect published genotyping and subtyping studies on Cryptosporidium spp. and Giardia duodenalis in the MENA region. These studies indicate that both anthroponotic and zoonotic transmission of Cryptosporidium occurs with the predominance of zoonotic transmission in most countries. Seven Cryptosporidium species were identified in humans (C. parvum, C. hominis, Cryptosporidium meleagridis, C. felis, Cryptosporidium muris, C. canis and C. bovis), with C. parvum by far being the most prevalent species (reported in 95.4% of the retrieved studies). Among C. parvum gp60 subtype families, IIa and IId predominated, suggesting potential zoonotic transmission. However, in four MENA countries (Lebanon, Israel, Egypt and Tunisia), C. hominis was the predominant species with five subtype families reported including Ia, Ib, Id, If and Ie, all of which are usually anthroponotically transmitted between humans. In animals, the majority of studies were conducted mainly on livestock and poultry, 15 species were identified (C. parvum, C. hominis, C. muris, Cryptosporidium cuniculus, C. andersoni, C. bovis, C. meleagridis, C. baileyi, C. erinacei, C. ryanae, C. felis, C. suis, Cryptosporidium galli, C. xiaoi and C. ubiquitum) with C. parvum (IIa and IId subtypes) the dominant species in livestock and C. meleagridis and C. baileyi the dominant species in poultry. With G. duodenalis, five assemblages (A, B, C, E and F) were identified in humans and six (A, B, C, E, D and F) in animals in MENA countries with assemblages A and B commonly reported in humans, and assemblages A and E dominant in livestock. This review also identified a major knowledge gap in the lack of Cryptosporidium spp. and Giardia duodenalis typing studies in water and food sources in the MENA region. Of the few studies conducted on water sources (including drinking and tap water), ten Cryptosporidium species and four genotypes were identified, highlighting the potential role of water as the major route of Cryptosporidium spp. transmission in the region. In addition, three G. duodenalis assemblages (A, B and E) were detected in different water sources with AI, AII and BIV being the main sub-assemblages reported. More research is required in order to better understand the molecular ersity and transmission dynamics of Cryptsporidum spp. and Giardia duodenalis in humans, animals, water and food sources in MENA region.
Publisher: Springer Vienna
Date: 13-09-2013
Publisher: Elsevier BV
Date: 10-2008
DOI: 10.1016/J.EXPPARA.2008.06.005
Abstract: Little is known about the prevalence of Isospora in domestic pigs in Western Australia. A total of 289 pig faecal s les were collected from pre- and post-weaned pigs and sows from 1 indoor and 3 outdoor piggeries located in the south-west region of Western Australia. Faecal s les were screened using a PCR-RFLP assay based on the ITS-1 rDNA locus. An overall prevalence of 10.4% (30/289) was identified. Isospora was detected in 16.3% (20/123) of pre-weaned animals and 6.4% (10/156) of post-weaned animals. PCR-RFLP analysis confirmed the presence of Isospora suis in 86.7% of the positive Isospora isolates. Isospora was significantly associated with diarrhea and the findings of this study suggest that management factors such as cleaning practices, flooring types and stocking densities need to be investigated in the porcine host to find new and improved measures for control.
Publisher: Cambridge University Press (CUP)
Date: 11-10-2013
DOI: 10.1017/S0022149X12000594
Abstract: Gastrointestinal parasites of livestock cause diseases of important socio-economic concern worldwide. The present study investigated the prevalence of gastrointestinal parasites in sheep and goats in lowland and highland regions of Papua New Guinea (PNG). Faecal s les were collected from a total of 165 small ruminants (110 sheep and 55 goats) from February to April 2011. Analysis by a modified McMaster technique revealed that 128 animals (72% of sheep and 89% of goats) were infected with one or more species of gastrointestinal parasites. The gastrointestinal parasites found and their prevalences in sheep (S) and in goats (G) were as follows: strongyle 67.3% (S), 85.5% (G) Eimeria 17.3% (S), 16.4% (G) Strongyloides , 8.2% (S), 23.6% (G) Fasciola , 5.5% (S), 18.2% (G) Trichuris , 1.8% (S), 3.6% (G) and Nematodirus , 1.8% (S), 3.6% (G). Two additional genera were found in goats: Moniezia (9.1%) and Dictocaulus (3.6%). This is the first study to quantitatively examine the prevalence of gastrointestinal parasites in goats in PNG. The high rates of parasitism observed in the present study are likely to be associated with poor farming management practices, including lack of pasture recovery time, lack of parasite control measures and poor-quality feed.
Publisher: Elsevier BV
Date: 09-2018
DOI: 10.1016/J.IJPARA.2018.05.002
Abstract: Recent molecular and sero-surveillance studies of the tick-borne pathogen Hepatozoon canis have identified new hosts, potential vector species, and have revealed that H. canis is more widespread than previously thought. We report the first diagnosed case of canine hepatozoonosis in Australia from a Maremma Sheepdog in Sarina, Queensland. Hepatozoon canis was detected with blood smear examination and 18S rRNA sequencing. It is unknown when or how the organism was introduced into Australia, which raises questions about border biosecurity policies and the H. canis infection status of its potential vectors and hosts in Australia. Surveillance for this pathogen is required to determine whether H. canis has established in Australia.
Publisher: Elsevier BV
Date: 04-2018
DOI: 10.1016/J.PREVETMED.2018.02.004
Abstract: Associations between faecal shedding of pathogenic Yersinia enterocolitica (based on the yst virulence gene) with growth, carcass weight and diarrhoea were investigated using an observational longitudinal study of 1200 crossbred prime (meat) lambs on eight Australian farms. Live weight, breech faecal soiling score (scale 1-5) and faecal consistency score (FCS scale 1-5) were recorded, and faecal s les collected from each lamb on three s ling occasions weaning (≈12 weeks of age), post-weaning (≈19 weeks) and pre-slaughter (≈29 weeks). Hot standard carcass weight was measured at slaughter. Faecal s les were screened for presence and concentration of pathogenic Y. enterocolitica using quantitative PCR. Associations of pathogenic Y. enterocolitica detection and shedding intensity with lamb health and production were assessed using general linear models (carcass weight), linear mixed effects models (live weight, FCS and breech soiling score) and non-parametric tests (FCS and breech soiling score). Prevalence for non-pelleted faeces (FCS ≥ 3.0) and diarrhoea (FCS ≥ 4.0) were compared with the two-tailed z-test, odds ratios and relative risk. Lambs shedding pathogenic Y. enterocolitica were 3.78 kg lighter post-weaning (P < 0.001) and 2.61 kg lighter pre-slaughter (P = 0.035) compared to lambs in which pathogenic Y. enterocolitica was not detected. Higher faecal concentration of pathogenic Y. enterocolitica was associated with lower live weight (P < 0.001). There was no association between pathogenic Y. enterocolitica detection and carcass weight. Overall, there was no evidence of association between pathogenic Y. enterocolitica detection and diarrhoea (higher FCS, higher risk for non-pelleted faeces or diarrhoea, or higher breech soiling score). Only one flock had increased relative risk for non-pelleted faeces associated with pathogenic Y. enterocolitica detection, and one other flock had increased relative risk for diarrhoea associated with pathogenic Y. enterocolitica detection. This is the first report of an association between reduced sheep live weight and pathogenic Y. enterocolitica based on the presence of the yst gene for heat stable enterotoxin determined by qPCR in sheep. Notably, impacts on live weight were observed in the absence of diarrhoea.
Publisher: Elsevier BV
Date: 09-2016
DOI: 10.1016/J.VETPAR.2016.08.003
Abstract: Associations between intensity and frequency of Cryptosporidium and Giardia shedding with growth, carcase weight and dressing% were investigated using a longitudinal study of 1182 lambs on eight Australian farms. Live weight was recorded and faecal s les were collected on three s ling occasions weaning (approximately 12 weeks of age), post-weaning (approximately 19 weeks) and pre-slaughter (approximately 29 weeks). Hot standard carcase weight (HSCW) and dressing% were measured at slaughter. Faecal s les were screened for presence and concentration of Cryptosporidium, Giardia and Haemonchus oocysts using a quantitative PCR. Trichostrongylid eggs were quantified with modified McMaster faecal worm egg count (WEC). Protozoan shedding intensity was categorised as high (above median oocyst concentration in positive sheep), low (below median oocyst concentration in positive sheep) or not detected. Shedding was also categorised for shedding type (no shedding, single Giardia infection, single Cryptosporidium infection, concurrent Giardia and Cryptosporidium infection) and lambs were categorised for frequency of shedding (shedding identified on 0, 1, 2 or 3 occasions). Associations of parasite shedding intensity category, shedding type, shedding frequency, WEC and Haemonchus status (positive or negative) with lamb production were assessed using general linear models (HSCW and dressing%) and linear mixed effects models (live weight). High Cryptosporidium parvum shedding was associated with lower live weight, ranging 2.31-4.52kg over the 3 s ling occasions. Cryptosporidium parvum shedding was associated with less HSCW in high (3.22kg less) and low (3.22kg less) shedding lambs post-weaning, and high (2.21kg less) and low (2.60kg less) shedding lambs pre-slaughter as well as lower dressing% (2.7% lower in high shedding lambs post-weaning). Cryptosporidium (all species) shedding pre-slaughter was associated with reduced dressing% in both high (1.25% lower) and low (1.21% lower) shedding lambs. Giardia shedding pre-slaughter was associated with 0.59kg less HSCW in high shedding lambs. Increased frequency of C. parvum and Giardia shedding in a specific animal (repeated detection) were associated with reduced HSCW and dressing%. Concurrent Giardia and Cryptosporidium shedding pre-slaughter was associated with reduced dressing%. No statistically significant main effects for either WEC (P>0.05) or Haemonchus status (P>0.05) were identified for any of the sheep meat productivity measures (live weight, HSCW and dressing%). The findings suggest naturally acquired Cryptosporidium and Giardia infections in grazing sheep are associated with depressed growth, carcase weight and dressing efficiency beyond the neonatal period in sheep representing a range of genetic backgrounds and different sheep production environments.
Publisher: Public Library of Science (PLoS)
Date: 13-07-2017
Publisher: Wiley
Date: 26-06-2017
DOI: 10.1111/AVJ.12597
Publisher: Wiley
Date: 17-01-2021
DOI: 10.1111/ZPH.12806
Publisher: Elsevier BV
Date: 2008
DOI: 10.1016/J.EXPPARA.2007.05.012
Abstract: To investigate the molecular epidemiology of Cryptosporidium species in children in Australia, fecal specimens from 50 Australian children with gastrointestinal symptoms and seven isolates from Australian neonatal dairy calves were genotyped and sub-genotyped at the 18S rDNA and GP60 loci, respectively, and compared with human and animal isolates collected from Europe, the US and Canada (n=35). Results revealed that the majority of the Australian human isolates were infected with C. hominis (41/50), while the remainder were infected with C. parvum. All the Australian cattle as well as cattle from US, Canada, UK and Switzerland were infected with C. parvum. Subtyping of 92 Cryptosporidium isolates at the GP60 locus identified seven subtype families of which six were identified in Australian isolates four C. hominis subtypes and two C. parvum subtypes. Results suggest that although transmission is largely anthroponotic in Australia, cattle may be a source of sporadic human infections.
Publisher: Elsevier BV
Date: 12-2006
DOI: 10.1016/J.VETPAR.2006.07.013
Abstract: A nested PCR that successfully detected Neospora caninum DNA in serum of cattle was used for investigation of selected abortion cases and in a study of healthy pregnant cows at an abattoir. N. caninum DNA was not detected in serum from antibody positive dams that aborted due to N. caninum, but was present in serum of some antibody negative dams that aborted due to other causes. N. caninum DNA was also found in the serum of about half of the animals that aborted of undetermined cause, but was not detected in cow sera from two beef cattle herds in Western Australia with no recent history of abortion. In the abattoir study of 79 dams and their foetuses N. caninum DNA was found in serum of 3 dams and in material from 11 foetuses. The majority of the cows and all foetuses were antibody negative. Our findings suggest that there is no obvious relationship between the presence or absence of N. caninum DNA in serum and the presence of antibodies to N. caninum in dams, the presence of N. caninum DNA in foetuses or abortion due to N. caninum. This is the first report of the detection of N. caninum DNA in serum of cattle rather than the white blood cell fraction. It indicates the presence of free tachyzoites and/or parasite DNA in circulation. The results suggest that persistent infection in the absence of antibodies is a possible outcome of N. caninum infection. Infection of foetuses in the absence of antibodies supports the possibility of persistent infection due to immunotolerance to an early in utero infection. It is therefore important to test for N. caninum DNA as well as antibodies for the detection of exposed and/or infected animals. However, the presence or absence of N. caninum antibodies or DNA did not support nor exclude N. caninum as the cause of abortion. Additional criteria are required for a positive diagnosis of abortion caused by N. caninum.
Publisher: Elsevier BV
Date: 06-2010
DOI: 10.1016/J.EXPPARA.2010.01.011
Abstract: Little is known about the epidemiology of Cryptosporidium in Jordan and no genotyping studies have been conducted on Cryptosporidium isolates from humans or animals from Jordan. Genotyping of 44 Cryptosporidium isolates from Jordanian children at the 18S rRNA locus and a unique diagnostic locus identified four Cryptosporidium species C. parvum (22), C. hominis (20), C. meleagridis (1) and C. canis (1). Sub-genotype analysis of 29 isolates at the 60-kDa glycoprotein (GP60) locus identified three C. parvum, two C. hominis subtype families and one C. meleagridis subtype. Several rare and novel subtypes were identified indicating unique endemicity and transmission of Cryptosporidium in Jordan.
Publisher: Elsevier BV
Date: 06-2014
DOI: 10.1016/J.EXPPARA.2014.03.021
Abstract: Species of Cryptosporidium are extensively recognised as pathogens of domesticated livestock and poultry, companion animals, wildlife, and are a threat to public health. Little is known of the prevalence of Cryptosporidium spp. in humans, domesticated animals or wildlife in Papua New Guinea (PNG). The aim of the present study was to screen sheep and goats for Cryptosporidium using molecular tools. A total of 504 faecal s les were collected from sheep (n=276) and goats (n=228) in village, government and institutional farms in PNG. S les were screened by nested PCR and genotyped at the 18S rRNA and at the 60kDa glycoprotein (gp60) loci. The overall prevalences were 2.2% for sheep (6/278) and 4.4% (10/228) for goats. The species/genotypes identified were Cryptosporidium hominis (subtype IdA15G1) in goats (n=6), Cryptosporidium parvum (subtypes IIaA15G2R1and IIaA19G4R1) in sheep (n=4) and in goats (n=2), Cryptosporidium andersoni (n=1) and Cryptosporidium scrofarum (n=1) in sheep, Cryptosporidium xiao (n=1) and Cryptosporidium rat genotype II (n=1) in goats. This is the first report of Cryptosporidium spp. identified in sheep and goats in PNG. Identification of Cryptosporidium in livestock warrants better care of farm animals to avoid contamination and illness in vulnerable population. The detection of zoonotic Cryptosporidium in livestock suggests these animals may serve as reservoirs for human infection.
Publisher: Elsevier BV
Date: 02-2014
DOI: 10.1016/J.VETPAR.2013.11.014
Abstract: The prevalence of Cryptosporidium in sheep in the eastern states of Australia has not been well described, therefore a study of the prevalence, oocyst concentration, species and subtypes of Cryptosporidium were assessed from lamb faecal s les at three s ling periods (weaning, post-weaning and pre-slaughter) from eight farms across South Australia, New South Wales, Victoria and Western Australia. A total of 3412 faecal s les were collected from approximately 1182 lambs across the four states and screened for the presence of Cryptosporidium using a quantitative PCR (qPCR) at the actin locus. Positives were typed at the 18S locus and at a second locus using C. parvum and C. hominis specific qPCR primers. The overall prevalence was 16.9% (95% CI: 15.6-18.1%) and of the 576 positives, 500 were successfully genotyped. In general, the prevalence of Cryptosporidium was higher in WA than the eastern states. Cryptosporidium prevalence peaked at 43.9% and 37.1% at Pingelly (WA2) and Arthur River (WA1), respectively during weaning and at Pingelly (WA2) during pre-slaughter (36.4%). The range of oocyst shedding at weaning overall across all states was 63-7.9×10(6) and the median was 3.2 × 10(4) oocysts g(-1). The following species were identified C. xiaoi (69%-345/500), C. ubiquitum (17.6%-88/500), C. parvum (9.8%-49/500), C. scrofarum (0.8%-4/500), mixed C. parvum and C. xiaoi (2.4%-12/500), C. andersoni (0.2%-1/500) and sheep genotype 1 (0.2%-1/500). Subtyping of C. parvum and C. ubiquitum isolates identified IIa and IId subtype families within C. parvum (with IId as the dominant subtype) and XIIa within C. ubiquitum. This is the first published description of C. parvum subtypes detected in lambs in Australia.
Publisher: Springer Science and Business Media LLC
Date: 11-03-2018
DOI: 10.1007/S00436-018-5808-8
Abstract: A new Isospora (Apicomplexa:Eimeriidae) species is described from a silvereye (Zosterops lateralis) in Western Australia. Sporulated oocysts of this species are spherical, 24.2 (23.1-25.2) × 23.3 (22.8-23.9) μm, with a shape index (length/width) of 1.02, and with a smooth bi-layered oocyst wall, 1.2 μm thick (outer layer 0.9 μm, inner 0.3 μm). A polar granule is present, but the oocyst residuum and a micropyle are absent. The ovoid-shaped sporocysts are 16.1 (15.7-17.3) × 10.5 (15.7-17.3) μm and have a shape index of 1.53. A hemidome-shaped Stieda and a rectangular-shaped substieda body are present. A sporocyst residuum is present and composed of numerous granules of different sizes scattered among the sporozoites. The oocysts from this isolate are morphologically different from those of all known Isospora spp. This coccidian parasite was molecularly characterised at the 18S, 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) gene. At the 18S locus, based on 1210 bp of sequence, this new isolate exhibited 99.9, 99.8, 99.7 and 99.5% similarity to I. sp. MAH-2013a (KF648870) from a superb starling (L rotornis superbus) in Canada, I. sp. MS-2003 (AY33157) from a Southern cape sparrow (Plocepasser mahali) in America, I. sp. Tokyo (AB75786) from Japan and I. sp. respectively. Further analysis of a subgroup of 300 bp long 18S sequences (n = 11), including I. anthochaerae and the other three Isospora characterised from birds in Western Australia, revealed that I. butcherae n. sp. exhibited 98.3% similarity to both I. sp. MAH-2013a (KF648870) and I. MS-2003 (AY33171). At the 28S locus, this new isolate exhibited 97.3% similarity with I. sp. MS-2003 from a California towhee (Melozone crissalis). At the COI locus, this new isolate exhibited 99.8% similarity to I. neochmiae from a red-browed finch. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora butcherae n. sp. after Mrs. June Butcher for her lifelong dedication as a wildlife rehabilitator. Graphical abstract ᅟ.
Publisher: Elsevier BV
Date: 04-2012
DOI: 10.1016/J.EXPPARA.2012.01.014
Abstract: Cryptosporidium is an enteric parasite of public health significance that causes diarrhoeal illness through faecal oral contamination and via water. Zoonotic transmission is difficult to determine as most species of Cryptosporidium are morphologically identical and can only be differentiated by molecular means. Transmission dynamics of Cryptosporidium in rural populations were investigated through the collection of 196 faecal s les from diarrheic (scouring) calves on 20 farms and 63 faecal s les from humans on 14 of these farms. The overall prevalence of Cryptosporidium in cattle and humans by PCR and sequence analysis of the 18S rRNA was 73.5% (144/196) and 23.8% (15/63), respectively. Three species were identified in cattle Cryptosporidium parvum, Cryptosporidium bovis and Cryptosporidium ryanae, and from humans, C. parvum and C. bovis. This is only the second report of C. bovis in humans. Subtype analysis at the gp60 locus identified C. parvum subtype IIaA18G3R1 as the most common subtype in calves. Of the seven human C. parvum isolates successfully subtyped, five were IIaA18G3R1, one was IIdA18G2 and one isolate had a mix of IIaA18G3R1 and IIdA19G2. These findings suggest that zoonotic transmission may have occurred but more studies involving extensive s ling of both calves and farm workers are needed for a better understanding of the sources of Cryptosporidium infections in humans from rural areas of Australia.
Publisher: Springer Science and Business Media LLC
Date: 23-11-2016
DOI: 10.1007/S00436-015-4831-2
Abstract: Cryptosporidium parvum commonly inhabits the intestinal tract of animals and humans and can cause acute watery diarrhea and weight loss. However, host immune responses to Cryptosporidium infections are not fully understood. IL-17 (also called IL-17A) is a pro-inflammatory cytokine of Th17 cells that plays a role in the host response to Cryptosporidium baileyi infection. The present study examined levels of IL-17-specific messenger RNA (mRNA) and Th17 associating cytokines in C. parvum-infected immune-suppressed BALB/c mice using real-time quantitative PCR (qPCR). Levels of IL-17 protein were determined by ELISA. The results showed that levels of IL-17 mRNA and Th17 cell-related cytokines, namely TGF-β, IL-6, STAT-3, RORγt, IL-22, TNF-α, and IL-23, were significantly increased (P < 0.05) in gut-associated lymphoid tissue (GALT) and spleen. IL-17 protein levels in GALT were also significantly increased (P < 0.05) after infection. The present study suggested that Th17 cells play a role in host-C. parvum interaction. These results could inform future studies of the immune response against C. parvum infection in transient immunosuppressed populations.
Publisher: Springer Science and Business Media LLC
Date: 20-03-2018
Publisher: Oxford University Press (OUP)
Date: 18-11-2010
DOI: 10.1111/J.1574-6968.2010.02134.X
Abstract: Cryptosporidium species generally lack distinguishing morphological traits, and consequently, molecular methods are commonly used for parasite identification. Various methods for Cryptosporidium identification have been proposed, each with their advantages and disadvantages. In this study, we show that capillary electrophoresis coupled with single-strand conformation polymorphism (CE-SSCP) is a rapid, simple and cost-effective method for the identification of Cryptosporidium species and genotypes. Species could be readily differentiated based on the SSCP mobility of lified 18S rRNA gene molecules. Clones that differed by single-nucleotide polymorphisms could be distinguished on CE-SSCP mobility. Profiles of species known to have heterogenic copies of 18S rRNA gene contained multiple peaks. Cloning and sequencing of Cryptosporidium parvum, Cryptosporidium hominis, Cryptosporidium fayeri and Cryptosporidium possum genotype 18S rRNA gene licons confirmed that these multiple peaks represented type A and type B 18S rRNA gene copies. CE-SSCP provides a reliable and sensitive analysis for epidemiological studies, environmental detection and ersity screening.
Publisher: American Society for Microbiology
Date: 2004
DOI: 10.1128/CMR.17.1.72-97.2004
Abstract: There has been an explosion of descriptions of new species of Cryptosporidium during the last two decades. This has been accompanied by confusion regarding the criteria for species designation, largely because of the lack of distinct morphologic differences and strict host specificity among Cryptosporidium spp. A review of the biologic species concept, the International Code of Zoological Nomenclature (ICZN), and current practices for Cryptosporidium species designation calls for the establishment of guidelines for naming Cryptosporidium species. All reports of new Cryptosporidium species should include at least four basic components: oocyst morphology, natural host specificity, genetic characterizations, and compliance with the ICZN. Altogether, 13 Cryptosporidium spp. are currently recognized: C. muris, C. andersoni, C. parvum, C. hominis, C. wrairi, C. felis, and C. cannis in mammals C. baïleyi, C. meleagridis, and C. galli in birds C. serpentis and C. saurophilum in reptiles and C. molnari in fish. With the establishment of a framework for naming Cryptosporidium species and the availability of new taxonomic tools, there should be less confusion associated with the taxonomy of the genus Cryptosporidium. The clarification of Cryptosporidium taxonomy is also useful for understanding the biology of Cryptosporidium spp., assessing the public health significance of Cryptosporidium spp. in animals and the environment, characterizing transmission dynamics, and tracking infection and contamination sources.
Publisher: Springer Science and Business Media LLC
Date: 24-04-2019
Publisher: Springer Science and Business Media LLC
Date: 21-07-2018
DOI: 10.1007/S00436-018-6017-1
Abstract: To identify the gastrointestinal helminths of veterinary, zoonotic and public health importance in farmers and their ruminant livestock in Ghana, faecal s les were collected from 95 farmers and their livestock (cattle = 328, sheep = 285 and goats = 217) and examined by microscopy and/or molecular techniques. Overall, 21 farmers tested positive for at least one gastrointestinal helminth, 80.9% of which were single infections and 19.0% co-infections. The parasites identified in the farmers consisted of hookworms (n = 13) (9 were Necator americanus and the other 4 could not be lified by PCR), Trichostrongylus spp. (n = 9), Schistosoma mansoni (n = 1), Schistosoma haematobium (n = 1) and Diphyllobothrium latum (n = 1). In livestock, strongylid nematodes were dominant (56.6%), followed by Par histomum spp. (16.9%), Dicrocoelium spp. (7.1%), Thysaniezia spp. (5.8%), Trichuris spp. (3.3%), Moniezia spp. (3.1%), Fasciola spp. (2.8%), Toxocara spp. (1.1%) and Schistosoma spp. (0.2%). Genotyping of Trichostrongylus spp. in the farmer's stools identified six T. colubriformis similar to T. colubriformis detected in cattle, sheep and goats in the study, two Trichostrongylus spp. with 98.3% and 99.2% genetic similarity to T. probolurus respectively and one Trichostrongylus spp. which showed 96.6% similarity to both T. probolurus and T. rugatus. Trichostrongylus axei was also identified in cattle, sheep and goats. This is the first molecular characterisation of Trichostrongylus spp. in Ghana and the species identified in the present study suggests zoonotic transmission from cattle, sheep and goats. Further studies involving larger numbers of farmers and their household members are essential to understand the transmission dynamics and impact of these parasites on farming communities in Ghana.
Publisher: Cambridge University Press (CUP)
Date: 11-08-2014
DOI: 10.1017/S0031182014001085
Abstract: Cryptosporidium is increasingly recognized as one of the major causes of moderate to severe diarrhoea in developing countries. With treatment options limited, control relies on knowledge of the biology and transmission of the members of the genus responsible for disease. Currently, 26 species are recognized as valid on the basis of morphological, biological and molecular data. Of the nearly 20 Cryptosporidium species and genotypes that have been reported in humans, Cryptosporidium hominis and Cryptosporidium parvum are responsible for the majority of infections. Livestock, particularly cattle, are one of the most important reservoirs of zoonotic infections. Domesticated and wild animals can each be infected with several Cryptosporidium species or genotypes that have only a narrow host range and therefore have no major public health significance. Recent advances in next-generation sequencing techniques will significantly improve our understanding of the taxonomy and transmission of Cryptosporidium species, and the investigation of outbreaks and monitoring of emerging and virulent subtypes. Important research gaps remain including a lack of subtyping tools for many Cryptosporidium species of public and veterinary health importance, and poor understanding of the genetic determinants of host specificity of Cryptosporidium species and impact of climate change on the transmission of Cryptosporidium.
Publisher: Elsevier BV
Date: 06-2012
DOI: 10.1016/J.VETPAR.2012.01.007
Abstract: Faecal s les (n=1155) were collected from n=111 (Farm A) and n=124 (Farm B) 2-6 week old female lambs on two farms in southern Western Australia across five s ling occasions (spanning 8 months). Genomic DNA was extracted directly from faecal s les and screened by PCR for ITS-2 nuclear ribosomal DNA to detect patent strongylid infections, specifically Teladorsagia circumcincta, Trichostrongylus spp., Haemonchus contortus, Oesophagostomum spp. and Chabertia ovina. The minimum amount of extracted genomic DNA necessary for successful PCR lification was 2.0-5.0 pg. During the five s ling occasions for the two farms, the sensitivities for WEC and PCR identification of strongylid infections varied, with levels of agreement between the two sets of diagnostic results ranging from 85 to 100%. Strongylid species prevalences were high (90.3-97.3%), with T. circumcincta and Trichostrongylus spp. the most prevalent species and together they were the most common mixed strongylid infection H. contortus was not identified in either flock. T. circumcincta was the only species associated with an increased risk of non-pelleted faeces on Farm B, where T. circumcincta-positive lambs were 2.3 and 2.6 times more likely to have non-pelleted faeces than negative lambs at the second and final s lings, respectively. The highest strongylid prevalence, mixed strongylid prevalence and mean number of strongylid species detected per lamb coincided with the highest average flock faecal worm egg counts (WECs) on both farms. There was a positive correlation between the number of strongyle species detected per lamb and both WEC and adjusted WEC (P<0.01 r(2) 0.026-0.591). These results indicate that strongylid eggs were likely to be the main source of strongylid DNA in the faecal DNA extracts. Despite the progress made by the molecular approach utilised in this study, it is incapable of distinguishing between patent and non-patent sources of strongylid DNA. However there is potential for further investigation into the development of a similar molecular procedure which could be used for early larvae detection on pastures.
Publisher: Elsevier BV
Date: 05-2015
DOI: 10.1016/J.MOLBIOPARA.2015.05.002
Abstract: Giardia duodenalis assemblage B is potentially a zoonotic parasite. The characterisation and investigation of isolates has been h ered by greater genetic ersity of assemblage B, limiting the application and utility of current genotyping loci. Since whole genome sequencing is the optimal high-throughput method for gene identification, the present study sequenced assemblage B isolate BAH15c1 and compared the sequence to the draft GS references to identify polymorphic genes for potential use in genotyping assays. The majority of the genome sequence was conserved between the two isolates, producing 508 contigs of 10.4 Mb with 4968 genes. Seventy polymorphic genes for potential use in genotyping assays were identified ranging in variation from elongation factor 1 α, which was the most conserved, through to triose phosphate isomerase, which was the most variable.
Publisher: Elsevier BV
Date: 05-2017
Publisher: Springer Science and Business Media LLC
Date: 1993
DOI: 10.1007/BF00931216
Publisher: Cambridge University Press (CUP)
Date: 07-2009
DOI: 10.1017/S0031182009006313
Abstract: The morphology and genetic characterization of a new species of trypanosome infecting koalas ( Phascolarctos cinereus ) are described. Morphological analysis of bloodstream forms and phylogenetic analysis at the 18S rDNA and gGAPDH loci demonstrated this trypanosome species to be genetically distinct and most similar to Trypanosoma bennetti , an avian trypanosome with a genetic distance of 0·9% at the 18S rDNA and 10·7% at the gGAPDH locus. The trypanosome was detected by 18S rDNA PCR in the blood s les of 26 out of 68 (38·2%) koalas studied. The aetiological role of trypanosomes in koala disease is currently poorly defined, although infection with these parasites has been associated with severe clinical signs in a number of koalas. Based on biological and genetic characterization data, this trypanosome species infecting koalas is proposed to be a new species Trypanosome irwini n. sp.
Publisher: Elsevier BV
Date: 11-2011
DOI: 10.1016/J.MOLBIOPARA.2011.07.007
Abstract: Patent strongylid nematode infections were identified using McMaster worm egg counts (WEC) and PCR assays (ITS-2 nuclear ribosomal DNA) to screen genomic DNA extracted directly from lamb faecal s les. Lambs from four different farms in southern Western Australia were s led rectally on two separate occasions, with McMaster WECs and PCRs conducted on a total of 858 s les. Negative controls (n=96) (WEC <50 eggs per gram [epg]) and positive controls (n=96) (faecal s les spiked with a 100 μL suspension of third-stage larvae (L(3)) containing approximately equal proportions of Teladorsagia circumcincta, Trichostrongylus colubriformis, Haemonchus contortus, Oesophagostomum spp. and Chabertia ovina) were generated. All control s les lified in accordance with positive controls. High levels of agreement (Kappa values ≥ 0.93) were identified between the two diagnostic tests. PCRs detected an additional 2.0% of s les as strongylid-positive but there was no significant difference in the number of strongylid-positive s les identified using PCR or McMaster WEC.
Publisher: Elsevier BV
Date: 11-2019
DOI: 10.1016/J.MEEGID.2019.104021
Abstract: Giardia duodenalis is one of the most common intestinal parasites in humans as well as livestock and wildlife. It is of both public and veterinary health importance in developing nations. A molecular survey of Giardia duodenalis assemblages in ruminants from Yazd Province, Iran was conducted on 484 animal faecal s les collected per rectum from slaughtered ruminants including 192 cattle, 192 sheep and 100 goats from June to November 2017. Species-specific and assemblage-specific PCRs for assemblages A, B and E at the triose phosphate isomerase (tpi) gene were performed, and s les positive for Giardia were confirmed by sequencing. In total, 25 (5.16%) of examined faecal s les including eight cattle (4.2%), twelve sheep (6.2%) and five goats (5%) were infected with G. duodenalis. Assemblage-specific PCR detected G. duodenalis assemblage E in seven faecal s les (six in sheep and one in a goat). Assemblages A and B were not detected. This study provides the first insight into Giardia infection in slaughtered livestock in Iran. Although the prevalence of infection with Giardia in this hot-arid area of Iran was low, educating people about direct contact with livestock such as farmers and abattoirs workers about this zoonotic infection is important.
Publisher: Elsevier BV
Date: 06-2004
Publisher: Public Library of Science (PLoS)
Date: 24-01-2017
Publisher: Elsevier BV
Date: 09-2017
DOI: 10.1016/J.IJPARA.2017.03.003
Abstract: The extent of within-host genetic ersity of parasites has implications for our understanding of the epidemiology, disease severity and evolution of parasite virulence. As with many other species, our understanding of the within-host ersity of the enteric parasite Cryptosporidium is changing. The present study compared Sanger and Next Generation Sequencing of glycoprotein 60 (gp60) licons from Cryptosporidium hominis (n=11), Cryptosporidium parvum (n=22) and Cryptosporidium cuniculus (n=8) DNA s les from Australia and China. Sanger sequencing identified only one gp60 subtype in each DNA s le: one C. hominis subtype (IbA10G2) (n=11), four C. parvum subtypes belonging to IIa (n=3) and IId (n=19) and one C. cuniculus subtype (VbA23) (n=8). Next Generation Sequencing identified the same subtypes initially identified by Sanger sequencing, but also identified additional gp60 subtypes in C. parvum and C. cuniculus but not in C. hominis, DNA s les. The number of C. parvum and C. cuniculus subtypes identified by Next Generation Sequencing within in idual DNA s les ranged from two to four, and both C. parvum IIa and IId subtype families were identified within the one host in two s les. The finding of the present study has important implications for Cryptosporidium transmission tracking as well as vaccine and drug studies.
Publisher: Elsevier BV
Date: 08-2021
Publisher: Elsevier BV
Date: 06-2012
DOI: 10.1016/J.TVJL.2011.06.042
Abstract: Diarrhoea is a widespread problem for sheep enterprises worldwide. A cross-sectional epidemiological study was conducted using a questionnaire to determine the prevalence of diarrhoea and associated risk factors where there was evidence of recent diarrhoea (active diarrhoea or fresh faecal soiling of breech fleece) for meat lambs on farms in southern Western Australia during 2010. The response rate was 41.4% (139/336). Evidence of recent diarrhoea was reported on 64.8% of farms, with a mean of 6.9% lambs affected per farm. Location of a farm and a higher annual rainfall were associated with an increased diarrhoea prevalence. Binary logistic regression analysis suggested that the drinking water source was associated with the incidence of diarrhoea, since lamb flocks supplied with dam water were 117 times (95% CI: 18.2, 754.8) more likely to have observed diarrhoea or fresh breech fleece faecal soiling than lamb flocks supplied with other sources of water. Faecal worm egg counts were used by 65% of respondents to determine whether an anthelmintic treatment was warranted and 74% of respondents administered a treatment to their meat lambs. In response to a range of diarrhoea scenarios presented to respondents (5%, 25% and 50% of the flock with evidence of recent diarrhoea), 15.1% would have elected to administer an anthelmintic treatment regardless of differences in prevalence.
Publisher: Elsevier BV
Date: 03-2015
DOI: 10.1016/J.EXPPARA.2015.01.009
Abstract: The morphological, biological, and molecular characteristics of Cryptosporidium piscine genotype 1 from the guppy (Poecilia reticulata) are described, and the species name Cryptosporidium huwi n. sp. is proposed to reflect its genetic and biological differences from gastric and intestinal Cryptosporidium species. Oocysts of C.huwi n. sp. over-lap in size with Cryptosporidium molnari, measuring approximately 4.4-4.9 µm (mean 4.6) by 4.0-4.8 µm (mean 4.4 µm) with a length to width ratio of 1.04 (0.92-1.35) (n = 50). Similar to C.molnari, C.huwi n. sp. was identified in the stomach only and clusters of oogonial and sporogonial stages were identified deep within the epithelium. However, phylogenetic analysis of 18S rRNA sequences indicated that C. huwi n. sp. exhibited 8.5-9.2% and 3.5% genetic distance from C.molnari isolates and piscine genotype 7 respectively. At the actin locus, the genetic distance between C.huwi n. sp. and C.molnari was 16.6%. The genetic distance between C.huwi n. sp. and other Cryptosporidium species at the 18S locus was 13.2%-17% and at the actin locus was 18.9%-26.3%. Therefore C. huwi n. sp. is genetically distinct from previously described Cryptosporidium species.
Publisher: Elsevier BV
Date: 11-2010
DOI: 10.1016/J.VETPAR.2010.08.006
Abstract: Little is known about the prevalence and genotypes of Cryptosporidium in fish. The present study investigated the prevalence of Cryptosporidium species in 200 aquarium fish of 39 different species in Western Australia by PCR lification at the 18S rRNA locus. A total of 21 positives were detected by PCR (10.5% prevalence) from 13 different species of fish. Nineteen of these isolates were successfully sequenced. Of these, 12 were similar or identical to previously described species/genotypes of Cryptosporidium, while the remaining seven isolates appeared to represent three novel species.
Publisher: Elsevier BV
Date: 12-2021
Publisher: Elsevier BV
Date: 11-2019
DOI: 10.1016/J.ACTATROPICA.2019.105126
Abstract: Gastrointestinal (GIT) parasite infections result in significant economic losses to ruminant livestock production. To determine the prevalence and risk factors associated with GIT parasite infections in livestock from Ghana, a cross-sectional survey was conducted in cattle and small ruminants kept under different management systems in the Coastal Savannah zone from October 2014 to February 2015. Faecal s les were collected from 328 cattle and 502 small ruminants (sheep and goats) and examined by formal ether concentration microscopy. The management systems and environmental conditions of the farm or household were observed, and a questionnaire administered to the livestock owners. Overall, 90.8% (754/830) of livestock were infected with at least one of ten different parasites (Eimeria, Strongylid nematodes, Toxocara, Trichuris, Schistosoma, Dicrocoelium, Par histomum, Fasciola, Moniezia and Thysaniezia), with Eimeria the most prevalent (78.4%). Most (64.5%) livestock had coinfections with two to five parasites with parasite intensity mostly light and at least one parasite was found in 98.6% (140/142) of the herds. Binary logistic regression models were generated to assess the risk factors associated with infection. Earthen floor was positively associated with strongylid infection, multiple ruminant species with Par histomum infection and flock size (>25 animal) with Thysaniezia, Dicrocoelium and Fasciola infections. Separating young animals from older animals was negatively associated with Strongylid infection, feed supplementation with Thysaniezia infection and small ruminant species with Par histomum and Toxocara infections. The findings from this study suggests that good sanitation, proper husbandry practices and improved nutrition can improve livestock health and production in Ghana.
Publisher: Elsevier BV
Date: 10-2020
Publisher: Elsevier BV
Date: 02-2017
DOI: 10.1016/J.VETPAR.2017.01.013
Abstract: Uninucleated Entamoeba cysts measuring 7.3×7.7μm were detected in faecal s les collected from wild Rangeland goats (Capra hircus) after arrival at a commercial goat depot near Geraldton, Western Australia at a prevalence of 6.4% (8/125). Sequences were obtained at the 18S rRNA (n=8) and actin (n=5) loci following PCR lification. At the 18S locus, phylogenetic analysis grouped the isolates closest with an E. bovis isolate (FN666250) from a sheep from Sweden with 99% similarity. At the actin locus, no E. bovis sequences were available, and the isolates shared 94.0% genetic similarity with E. suis from a pig in Western Japan. This is the first report to describe the morphology and molecular characterisation of Entamoeba from Rangeland goats in Western Australia and the first study to produce actin sequences from E. bovis-like Entamoeba sp.
Publisher: Elsevier BV
Date: 12-1998
DOI: 10.1016/S0169-4758(98)01247-2
Abstract: In this article, Una Morgan and Andrew Thompson briefly review the latest information on polymerase chain reaction (PCR)detection of Cryptosporidium parvum in both clinical and environmental s les. Current detection methods for Cryptosporidium are cumbersome, time-consuming and lack sensitivity. A variety of PCR tests have been described recently in the literature and this article discusses the advantages and disadvantages of each new technique and their potential for future diagnosis of Cryptosporidium.
Publisher: Elsevier BV
Date: 03-2011
DOI: 10.1016/J.VETPAR.2010.10.056
Abstract: A total of 364 fecal specimens from randomly selected pre-weaned calves, aged up to 4 months, from 5 different farms in the south of Western Australia and 1 farm from New South Wales were screened for the presence of Cryptosporidium and Giardia using PCR. There were substantial differences in prevalence between the farms and the overall prevalence was 22.3% (81/364) and 26.9% (98/364) respectively for Cryptosporidium and Giardia. For Cryptosporidium, 70 positives were identified at the 18S locus. At a unique diagnostic locus, an additional 12 C. parvum positives were identified. Sequence analysis at the 18S ribosomal RNA locus was successful for 59 of the 70 positive isolates of these 14 were C. parvum, 28 were C. bovis, 15 were C. ryanae, 1 was pig genotype II and 1 was a mixed C. ryanae/C. parvum infection. Sub-typing analysis at the glycoprotein 60 (gp60) locus for 24 C. parvum isolates identified all as IIa 17 were A17G2R1, 1 was A18G3R1 and 6 were A20G3R1. For Giardia, 75 positives were identified at the 18S locus and an additional 23 positives were identified at the gdh locus. The majority of the isolates sequenced were assemblage E, however assemblage A and B and mixed A and E and A, B and E infections as well as the quenda genotype were identified. The findings of the present study indicate that pre-weaned calves are not an important source of zoonotic Giardia species in Australia but may be an important source of zoonotic Cryptosporidium.
Publisher: Springer Science and Business Media LLC
Date: 20-06-2019
DOI: 10.1007/S00436-019-06378-8
Abstract: A new Isospora (Apicomplexa: Eimeriidae) species is described from an Australian raven (Corvus coronoides) in Western Australia. Sporulated oocysts (n = 21) are ovoid, 21.2 (18.4-23.9) μm in length and 18.8 (16.9-20.6) μm in width, with a shape index of 1.13. The bi-layered oocyst wall is smooth and colourless, 1.2 μm thick. A polar granule and oocyst residuum is present, but the micropyle is absent. The sporocysts are ovoid-shaped, 16.3 (13.7-18.9) × 10.7 (8.4-12.9) μm, with a shape index (length/width) of 1.52. Stieda and substieda bodies are present, the Stieda body being small and hemidome-shaped and the substieda being indistinct. Each sporocyst with four vermiform sporozoites arranged head to tail. The sporozoites are crescent-shaped, 9.0 (8.9-9.2) × 2.7 (2.3-3.0) μm, with a shape index (length/width) of 3.33. The sporocyst residuum is present. The isolated oocysts had different morphological characteristics when compared with all known Isospora spp. The coccidian parasite was analysed at the 18S and 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) loci. At the 18S locus, I. coronoideae n. sp. exhibited 98.9% similarity to I. neochmiae from a captive-bred red-browed finch (KT224380) and Isospora sp. from domestic pigeons (Columba livia domestica) (AB757860), 98.5% similarity to I. gryphoni (AF080613) from an American goldfinch and 98.3% similarity to I. manorinae (KT224379) from a yellow-throated miner. At the 28S locus, it exhibited 95.4% and 94.8% similarity to I. manorinae (KT224381) and I. anthochaerae (KF766053), respectively. At the COI locus, it exhibited 99.8% and 99.7% similarity to I. butcherae (KY801687) and I. neochmiae (KT224378), respectively. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora coronoideae n. sp. after its host, the Australian raven (Corvus coronoides) (Passeriformes: Corvidae) (Linnaeus, 1758).
Publisher: Elsevier BV
Date: 02-2013
DOI: 10.1016/J.EXPPARA.2012.11.004
Abstract: A novel Eimeria sp. was identified in faeces collected from a King's skink (Egernia kingii) housed at the Kanyana Wildlife Rehabilitation Centre in Western Australia. Oocysts measure 17.0×15.0 μm with a length/width ratio (L/W) of 1.13. Phylogenetic analysis of 18S rRNA sequences indicated that the novel Eimeria sp. shared the highest genetic similarity to Eimeria antrozoi and Eimeria rioarribaensis from vespertilionid bats from North America (≥98.9%). At the COI locus, bat-derived sequences were not available and phylogenetic analysis placed the novel Eimeria sp. in a clade by itself and shared 98.8% similarity with the rodent-derived species E. falciformis and E. vermiformis. This suggests that the isolate from the King's skink's faeces was probably derived from a mammal, possibly a rodent or a bat.
Publisher: American Society for Microbiology
Date: 07-2013
DOI: 10.1128/JCM.00424-13
Abstract: This report describes a case of cryptosporidiosis from an immunocompetent patient from Perth, Western Australia, suffering from diarrhea and a spectrum of other symptoms. Molecular identification revealed that this patient was infected with three Cryptosporidium species— Cryptosporidium meleagridis , the Cryptosporidium mink genotype, and an unknown Cryptosporidium species.
Publisher: Elsevier BV
Date: 12-2012
DOI: 10.1016/J.VETPAR.2012.07.017
Abstract: A molecular procedure was developed to detect and quantify larvae of different strongylid parasite species recovered from pasture s les. Two lamb flocks (L and S) grazed separate paddocks with different natural larvae challenges (one low [Paddock L] and one high [Paddock S] challenge) on a commercial farm in Western Australia. Pasture s les were collected and analysed for larvae on 9 separate occasions from each paddock. Pregnant Merino ewes were s led on 3 separate occasions (2 pre-partum and 1 post-partum). Following lambing, 203 female crossbred lambs were identified, from which faecal s les were collected across five separate s lings. Lamb production and faecal attributes were recorded. Genomic DNA was extracted directly from lamb faeces, in addition to the genomic DNA extracts from strongylid larval species recovered from pastures. Faecal worm egg counts (FWECs) were undertaken. Species-specific qPCRs and conventional PCRs (ITS-2 nuclear ribosomal DNA) were used to screen s les for strongylid species (Teladorsagia circumcincta, Trichostrongylus spp., Haemonchus contortus, Chabertia ovina and Oesophagostomum venulosum). Negative correlations (r(2)>0.91) were found between qPCR C(q) values and log-transformed pasture larval counts for Trichostrongylus spp. and T. circumcincta. Moderate levels of agreement between pasture larval counts and qPCR results were observed (67%). A clear difference in pasture larval challenge levels was observed between the two flocks using both qPCR and conventional pasture larval counts. It is difficult to draw conclusions on the production performances of lambs from the two experimental flocks, as no further replicates were able to be conducted following this experiment. Flock L had higher dressing percentages than Flock S (P=0.038), along with significantly higher faecal consistency and breech fleece faecal soiling scores at successive s lings. The molecular procedures utilised in this study have the potential to be beneficial for livestock grazing management strategies and parasite surveillance, however further investigation is necessary before they can become part of routine diagnostics.
Publisher: Microbiology Society
Date: 16-12-2021
Abstract: Advances in sequencing technologies have revealed the complex and erse microbial communities present in ticks (Ixodida). As obligate blood-feeding arthropods, ticks are responsible for a number of infectious diseases that can affect humans, livestock, domestic animals and wildlife. While cases of human tick-borne diseases continue to increase in the northern hemisphere, there has been relatively little recognition of zoonotic tick-borne pathogens in Australia. Over the past 5 years, studies using high-throughput sequencing technologies have shown that Australian ticks harbour unique and erse bacterial communities. In the present study, free-ranging wildlife ( n =203), representing ten mammal species, were s led from urban and peri-urban areas in New South Wales (NSW), Queensland (QLD) and Western Australia (WA). Bacterial metabarcoding targeting the 16S rRNA locus was used to characterize the microbiomes of three s le types collected from wildlife: blood, ticks and tissue s les. Further sequence information was obtained for selected taxa of interest. Six tick species were identified from wildlife: Amblyomma triguttatum , Ixodes antechini , Ixodes australiensis , Ixodes holocyclus , Ixodes tasmani and Ixodes trichosuri . Bacterial 16S rRNA metabarcoding was performed on 536 s les and 65 controls, generating over 100 million sequences. Alpha ersity was significantly different between the three s le types, with tissue s les displaying the highest alpha ersity ( P .001). Proteobacteria was the most abundant taxon identified across all s le types (37.3 %). Beta ersity analysis and ordination revealed little overlap between the three s le types ( P .001). Taxa of interest included Anaplasmataceae , Bartonella , Borrelia , Coxiellaceae , Francisella , Midichloria , Mycoplasma and Rickettsia . Anaplasmataceae bacteria were detected in 17.7% (95/536) of s les and included Anaplasma , Ehrlichia and Neoehrlichia species. In s les from NSW, ‘ Ca . Neoehrlichia australis’, ‘ Ca . Neoehrlichia arcana’, Neoehrlichia sp. and Ehrlichia sp. were identified. A putative novel Ehrlichia sp. was identified from WA and Anaplasma platys was identified from QLD. Nine rodent tissue s les were positive for a novel Borrelia sp. that formed a phylogenetically distinct clade separate from the Lyme Borrelia and relapsing fever groups. This novel clade included recently identified rodent-associated Borrelia genotypes, which were described from Spain and North America. Bartonella was identified in 12.9% (69/536) of s les. Over half of these positive s les were obtained from black rats ( Rattus rattus ), and the dominant bacterial species identified were Bartonella coopersplainsensis and Bartonella queenslandensis . The results from the present study show the value of using unbiased high-throughput sequencing applied to s les collected from wildlife. In addition to understanding the sylvatic cycle of known vector-associated pathogens, surveillance work is important to ensure preparedness for potential zoonotic spillover events.
Publisher: Elsevier BV
Date: 2016
Publisher: Elsevier BV
Date: 03-2016
Publisher: Wiley
Date: 09-2016
DOI: 10.1111/PIM.12350
Abstract: Cryptosporidium is a major cause of moderate-to-severe diarrhoea in humans worldwide, second only to rotavirus. Due to the wide host range and environmental persistence of this parasite, cryptosporidiosis can be zoonotic and associated with foodborne and waterborne outbreaks. Currently, 31 species are recognized as valid, and of these, Cryptosporidium hominis and Cryptosporidium parvum are responsible for the majority of infections in humans. The immune status of the host, both innate and adaptive immunity, has a major impact on the severity of the disease and its prognosis. Immunocompetent in iduals typically experience self-limiting diarrhoea and transient gastroenteritis lasting up to 2 weeks and recover without treatment, suggesting an efficient host antiparasite immune response. Immunocompromised in iduals can suffer from intractable diarrhoea, which can be fatal. Effective drug treatments and vaccines are not yet available. As a result of this, the close cooperation and interaction between veterinarians, health physicians, environmental managers and public health operators is essential to properly control this disease. This review focuses on a One Health approach to prophylaxis, including the importance of understanding transmission routes for zoonotic Cryptosporidium species, improved sanitation and better risk management, improved detection, diagnosis and treatment and the prospect of an effective anticryptosporidial vaccine.
Publisher: Wiley
Date: 26-04-2017
DOI: 10.1111/AVJ.12572
Abstract: Develop a multiplex quantitative PCR assay to investigate the prevalence and shedding of Escherichia coli O157/O145, Salmonella spp. and C ylobacter spp. in sheep at sale yards and abattoirs. A qPCR for E. coli O157/O145 was developed, validated and multiplexed with an existing qPCR for C ylobacter and Salmonella enterica. The absolute numbers of E. coli O157/O145, C ylobacter and Salmonella in control s les was determined using droplet digital PCR. These were then used as the controls in the multiplex qPCR on a total of 474 sheep faecal s les collected from two saleyards over a 4-month period (April-July 2014) and 96 effluent s les from an abattoir. The mutiplex qPCR was specific with a sensitivity of 5 organisms/μL faecal DNA extract for C ylobacter, S. enterica and E. coli O157/O145. The overall prevalence of C ylobacter, S. enterica and E. coli O157/O145 in faecal s les was 5.7%, 3.6% and 8.4% and in effluent s les was 18.8%, 6.3% and 5.2%, respectively. The pathogen loads of C ylobacter, S. enterica and E. coli O157/O145 in faecal and effluent s les was also determined via mutiplex qPCR. The overall prevalences of C ylobacter, S. enterica and E. coli O157/O145 were generally low (<6%), but point prevalences ranged considerably in healthy sheep (up to 26% for E. coli O157/O145). Further work to determine risk factors for shedding of bacterial organisms in meat sheep in the pre-slaughter period (on-farm, sale yards and lairage at abattoirs) could further reduce the risk of contamination of meat products.
Publisher: Elsevier BV
Date: 2017
DOI: 10.1016/J.VPRSR.2016.11.006
Abstract: Faecal shedding of Cryptosporidium and Giardia by captured rangeland goats was investigated using a longitudinal study with four faecal s les collected from 125 male goats once monthly for four months, commencing immediately after capture and transport to a commercial goat depot (feedlot). Goats were composite breed and aged approximately 9-12months on arrival. Faecal s les were screened for Cryptosporidium and Giardia presence and concentration using quantitative PCR and sequencing at the 18S ribosomal RNA locus (Cryptosporidium), and glutamate dehydrogenase and β-giardin loci (Giardia). Longitudinal prevalence for Cryptosporidium was 27.2% (point prevalence range 3-14%) with 3 species identified: C. xiaoi (longitudinal prevalence 13.6%), C. ubiquitum (6.4%) and C. parvum (3.2%). Sub-typing at the gp60 locus identified C. ubiquitum XIIa, C. parvum IIaA17G2R1 and C. parvum IIaA17G4R1. This is the first report of the zoonotic C. parvum subtype IIaA17G4R1 in goats. The pattern of genotypes shed in faeces changed over the duration of study with C. ubiquitum identified only at the first and second s lings, and C. parvum identified only at the fourth s ling. Longitudinal prevalence for Giardia duodenalis was 29.6% (point prevalence range 4-12%) with all positives sub-typed as assemblage E. Only 2/125 goats were identified to be shedding Cryptosporidium or Giardia on more than one occasion. This is the first report of Cryptosporidium and Giardia genotypes in captured rangeland goats. Faecal shedding of zoonotic Cryptosporidium spp. and potentially zoonotic G.duodenalis has implications for food safety and effluent management. Keywords: Cryptosporidium Giardia Rangeland goats zoonotic.
Publisher: Springer Science and Business Media LLC
Date: 15-04-2019
DOI: 10.1007/S00436-019-06317-7
Abstract: The present study assessed the prevalence and morphology of Leucocytozoon podargii from wild tawny frogmouths (Podargus strigoides) in Western Australia (WA) and genetically characterised the cytochrome b gene (cyt b) of L. podargii in wild tawny frogmouths from WA and Queensland (QLD). The prevalence of L. podargii in wild tawny frogmouths from WA was 93.3% (14/15 95% CI, 68.1-99.8%). The morphological characters of L. podargii from WA were similar to L. podargii from QLD: the gametocytes were round-oval shape, approximately 8-12 μm in diameter the macrogametocytes were 12.4 μm in diameter microgametocytes were 10.4 μm in diameter and the ratio of macrogametocytes and microgametocytes was 3:2. Sequence analysis of partial cyt b gene fragments revealed that L. podargii sequences isolated from wild tawny frogmouths in WA shared the highest similarity (99.8% at nucleotide level and 100% at protein level) with L. podargii isolated from wild tawny frogmouths in QLD. The mitochondrial 18S rRNA gene of L. podargii gametocytes was quantified using droplet digital PCR (ddPCR), and the highest gametocyte load was detected in the lung. This finding corresponds to the results of the histological study. Based on the morphological and molecular studies, it was concluded that the Leucocytozoon parasite identified from wild tawny frogmouths in WA is consistent with L. podargii from wild tawny frogmouths in QLD, and the present study has genetically characterised two different L. podargii genotypes (QLD and WA) for the first time.
Publisher: Elsevier BV
Date: 06-2014
DOI: 10.1016/J.EXPPARA.2014.03.004
Abstract: In 2012, the first autochthonous Australian case of human babesiosis was reported, after microscopic examinations of blood s les revealed intra-erythrocytic parasites in a hospitalized 56year-old man from NSW, who died in 2011 (Senanayake et al., 2012). Independent molecular analyses carried out in Australia and the USA, identified Babesia microti at the 18S ribosomal RNA (18S rRNA), and the beta-tubulin (β-tubulin) gene loci. Here we present the details of a novel PCR-based assay for the β-tubulin gene that was developed, during the original study, to corroborate the results obtained from the analysis of the 18S rDNA. The complete phylogenetic reconstruction, based on the two loci sequenced from the Australian clinical isolate, is also shown here for the first time.
Publisher: Cambridge University Press (CUP)
Date: 06-2002
DOI: 10.1017/S0031182002002366
Abstract: Since isolates of Hymenolepis nana infecting humans and rodents are morphologically indistinguishable, the only way they can be reliably identified is by comparing the parasite in each host using molecular tools. In the current study, isolates of H. nana from rodent and human hosts from a broad geographical range were sequenced at the ribosomal first internal transcribed spacer (ITS1), the mitochondrial cytochrome c oxidase subunit 1 (C01) gene and the nuclear paramyosin gene loci. Twenty-three isolates of H. nana were sequenced at the ITS1 locus and this confirmed the existence of spacers which, although similar in length (approximately 646 bp), differed in their primary sequences which led to the separation of the isolates into 2 clusters when analysed phylogenetically. This sequence variation was not, however, related to the host of origin of the isolate, thus was not a marker of genetic distinction between H. nana from rodents and humans. Sequencing of a 444 bp fragment of the mitochondrial cytochrome c oxidase 1 gene (C01) in 9 isolates of H. nana from rodents and 6 from humans identified a phylogenetically supported genetic ergence of approximately 5% between some mouse and human isolates. This suggests that H. nana is a species complex, or 'cryptic' species (=morphologically identical yet genetically distinct). A small segment of the nuclear gene, paramyosin, (625 bp or 840 bp) was sequenced in 4 mouse and 3 human isolates of H. nana. However, this gene did not provide the level of heterogeneity required to distinguish between isolates from rodent and human hosts. From the results obtained from faster evolving genes, and the epidemiological evidence, we believe that the life-cycle of H. nana that exists in the north-west of Western Australia is likely to involve mainly 'human to human' transmission.
Publisher: Cambridge University Press (CUP)
Date: 27-04-2011
DOI: 10.1017/S0031182011000369
Abstract: Whole blood collected from koalas admitted to the Australian Zoo Wildlife Hospital (AZWH), Beerwah, QLd, Australia, during late 2006–2009 was tested using trypanosome species-specific 18S rDNA PCRs designed to lify DNA from Trypanosoma irwini, T. gilletti and T. copemani . Clinical records for each koala s led were reviewed and age, sex, blood packed cell volume (PCV), body condition, signs of illness, blood loss, trauma, chlamydiosis, bone marrow disease, koala AIDS and hospital admission outcome (‘survival’ / ‘non-survival’) were correlated with PCR results. Overall 73·8% (439/595) of the koalas were infected with at least 1 species of trypanosome. Trypanosoma irwini was detected in 423/595 (71·1%), T. gilletti in 128/595 (21·5%) and T. copemani in 26/595 (4·4%) of koalas. Mixed infections were detected in 125/595 (21%) with co-infections of T. irwini and T. gilletti (101/595, 17%) being most common. There was a statistical association between infection with T. gilletti with lower PCV values and body condition scores in koalas with signs of chlamydiosis, bone marrow disease or koala AIDS. No association between T. gilletti infection and any indicator of health was observed in koalas without signs of concurrent disease. This raises the possibility that T. gilletti may be potentiating other disease syndromes affecting koalas.
Publisher: Elsevier BV
Date: 09-2013
DOI: 10.1016/J.EXPPARA.2013.06.014
Abstract: Cryptosporidium is an enteric protozoan parasite that is resistant to inactivation by commonly used drinking water disinfectants. Between 2004 and 2010, it was responsible for 60% of all waterborne protozoan parasitic outbreaks reported worldwide. Most sporadic infections in humans and almost all outbreaks are caused by Cryptosporidium parvum and Cryptosporidium hominis. We report the development and validation of a quantitative qPCR assay using minor groove binder (MGB)-probes targeting a unique Cryptosporidium specific protein-coding gene, that directly detects, quantitates and identifies C. hominis and C. parvum in environmental and faecal s les. An internal lification control (IAC) was also developed and included in this assay. The qPCR assay was compared with an 18S nested PCR assay for sensitivity and specificity. The analytical sensitivity for the qPCR assay was 1 oocyst and 1-10 oocysts for the 18S assay. Evaluation of analytical specificity of the qPCR assay revealed no cross-reactions with other genera and detected all C. parvum and C. hominis isolates correctly. The diagnostic sensitivity and specificity of the qPCR was 100% compared to 96.9% and 98.4%, respectively for the 18S assay. The qPCR assay was also highly reproducible with RSD (relative standard deviation) values of 1.4-9.4%, when the assay was performed by four different technicians. When tested on water s les, the qPCR assay was more sensitive than the 18S assay, detecting positives in 37 of 138 water s les compared to 35 for the 18S locus. This qPCR assay should be a valuable tool for the detection and differentiation of C. hominis and C. parvum in both clinical and environmental s les.
Publisher: Elsevier BV
Date: 02-2014
DOI: 10.1016/J.EXPPARA.2013.12.004
Abstract: A novel quantitative PCR (qPCR) for Giardia at the glutamate dehydrogenase (gdh) locus was developed and validated. The qPCR was used to screen a total of 3412 lamb faecal s les collected from approximately 1189 lambs at three s ling periods (weaning, post-weaning and pre-slaughter) from eight farms across South Australia (SA), New South Wales (NSW), Victoria (Vic) and Western Australia (WA). The overall prevalence was 20.2% (95% CI 18.9-21.6) and of the 690 positives, 473 were successfully typed. In general, the prevalence of Giardia varied widely across the different farms with the highest prevalence in one WA farm (42.1%) at pre-slaughter s ling and the lowest prevalence in one Victorian farm (7.2%) at weaning. The range of cyst shedding at weaning, post-weaning and pre-slaughter overall across all states was 63-1.3×10(9) cysts g(-1) (median=1.7×10(4)), 63-1.1×10(9) cysts g(-1) (median=9.6×10(3)), 63-4.7×10(9) cysts g(-1) (median=8.1×10(4)) respectively. Assemblage specific primers at the triose phosphate isomerase (tpi) locus identified assemblage A in 22.4% (106/473) of positive s les typed, assemblage E in 75.9% (359/473) and mixed A and E assemblages in 1.7% (8/473) of s les. A subset of representative s les from the 8 farms (n=32) were typed at both the gdh and beta-giardin loci and confirmed these results and identified sub-assemblage AII in 16 representative assemblage A isolates across the 8 farms. This demonstrates a prevalence of Giardia previously not recognised in Australian sheep, highlighting a need for further research to quantify the production impacts of this protozoan parasite.
Publisher: Elsevier BV
Date: 11-1999
DOI: 10.1016/S0020-7519(99)00109-5
Abstract: Cryptosporidium is an important cause of enteric disease in humans and other animals. Limitations associated with conventional diagnostic methods for cryptosporidiosis based on morphological features, coupled with the difficulty of characterising parasites isolated in the laboratory, have restricted our ability to clearly identify species. The application of sensitive molecular approaches has obviated the necessity for laboratory lification. Such studies have found considerable evidence of genetic heterogeneity among isolates of Cryptosporidium from different species of vertebrate, and there is now mounting evidence suggesting that a series of host-adapted genotypes/strains/species of the parasite exist. In this article, studies on the molecular characterisation of Cryptosporidium during the last 5 years are reviewed and put into perspective with the past and present taxonomy of the genus. The predictive value of achieving a sound taxonomy for the genus Cryptosporidium with respect to understanding its epidemiology and transmission and controlling outbreaks of the disease is also discussed.
Publisher: Cambridge University Press (CUP)
Date: 26-04-2011
DOI: 10.1017/S0031182011000497
Abstract: A total of 41 ticks were collected from 15 quokkas on Bald Island and 2 ticks from a Gilbert's potoroo from Two Peoples Bay. Three species of Ixodid ticks Ixodes australiensis, Ixodes hirsti and Ixodes myrmecobii were identified on the quokkas known to have a high prevalence of Trypanosoma copemani . Tick faeces from ticks isolated from 8 in idual quokkas and a Gilbert's potoroo were examined with one identified as positive for trypanosomes. Faecal examination revealed trypanosomes similar to in vitro life-cycle stages of T. copemani . In total 12 ticks were dissected and trypanosomes found in sections of their midgut and haemolymph, 49 and 117 days after collection. Tick faeces, salivary glands and midguts from I. australiensis were screened using an 18S rRNA PCR with lification seen only from the midguts. Sequencing showed 100% homology to T. copemani (genotype A) and 99·9% homology to the wombat (AII) isolate of T. copemani . Trypanosomes were only detected in I. australiensis as neither I. hirsti nor I. myrmecobii survived the initial 30-day storage conditions. We therefore identify a vector for T. copemani as I. australiensis and, given the detection of trypanosomes in the faeces, suggest that transmission is via the faecal-oral route.
Publisher: Elsevier BV
Date: 12-2002
DOI: 10.1016/S0020-7519(02)00197-2
Abstract: To assess the genetic ersity and evolution of Cryptosporidium parasites, the partial ssrRNA, actin, and 70kDa heat shock protein (HSP70) genes of 15 new Cryptosporidium parasites were sequenced. Sequence data were analysed together with those previously obtained from other Cryptosporidium parasites (10 Cryptosporidium spp. and eight Cryptosporidium genotypes). Results of this multi-locus genetic characterisation indicate that host adaptation is a general phenomenon in the genus Cryptosporidium, because specific genotypes were usually associated with specific groups of animals. On the other hand, host-parasite co-evolution is also common in Cryptosporidium, as closely related hosts usually had related Cryptosporidium parasites. Results of phylogenetic analyses suggest that the Cryptosporidium parvum bovine genotype and Cryptosporidium meleagridis were originally parasites of rodents and mammals, respectively, but have subsequently expanded their host ranges to include humans. Understanding the evolution of Cryptosporidium species is important not only for clarification of the taxonomy of the parasites but also for assessment of the public health significance of Cryptosporidium parasites from animals.
Publisher: Springer Science and Business Media LLC
Date: 21-11-2020
DOI: 10.1007/S00436-019-06547-9
Abstract: The Bellinger River snapping turtle (Myuchelys georgesi) is endemic to Australia and is confined to a highly restricted distribution in the Bellinger River in New South Wales. Routine veterinary health examinations of 17 healthy turtles were undertaken, along with the collection and analysis of blood s les, during conservation efforts to save the species following a catastrophic population decline. Microscopy analysis of blood films detected Haemoproteidae parasites that morphologically resembled Haemocystidium chelodinae inside turtle erythrocytes. Of the 17 turtles examined, 16 were positive for infection with H. chelodinae by both light microscopy and PCR. DNA sequencing of a partial fragment of the mitochondrial cytochrome b (cytb) gene and phylogenetic analysis identified two different H. chelodinae-like genotypes. The phylogenetic relationship of H. chelodinae-like to other Haemoproteidae species based on cytb sequences grouped H. chelodinae-like into the reptile clade, but revealed the Haemocystidium genus to be paraphyletic as the clade also contained Haemoproteus, thus supporting a re-naming of Haemoproteus species from reptiles to Haemocystidium species. This study reports for the first time the genetic characterisation of H. chelodinae-like organisms isolated from a new Testudine host species, the Bellinger River snapping turtle. As evidence grows, further research will be necessary to understand the mode of transmission and to investigate whether these parasites are pathogenic to their hosts.
Publisher: Centers for Disease Control and Prevention (CDC)
Date: 05-2009
Publisher: Elsevier BV
Date: 04-2011
DOI: 10.1016/J.IJPARA.2010.11.007
Abstract: Recent research concerning Giardia duodenalis has focused on resolving possible sub-assemblages within Assemblages A and B to better understand host-specific and zoonotic relationships. In the present study nine cloned, cultured, Assemblage B isolates were used to investigate the intra-Assemblage B substitution patterns of conserved (ssrDNA, ef, h2b, h4) and variable (tpi, gdh, bg) genes to assess their suitability for further application to sub-assemblage analyses. The resolution of each gene was found to be proportional to its substitution rate and for the genetically narrow s le set examined, the variable genes best represented the consensus phylogeny while the conserved genes only established fractions. However it was demonstrated that the spectra of conserved and variable genes were required to ensure accuracy of inferred phylogeny and it was therefore concluded that further research into sub-Assemblage B groups would require a mixture of conserved and variable genes for the multi-locus analyses of this genetically broad assemblage.
Publisher: Elsevier BV
Date: 09-2011
DOI: 10.1016/J.VETPAR.2011.05.004
Abstract: An intestinal Eimeria was previously reported as a significant pathogen of Asian seabass (Lates calcarifer) in nurseries in Vietnam. In the present study, both Eimeria and Cryptosporidium were detected by sequence analyses of fragments of the 18S rRNA gene lified from these Vietnamese L. calcarifer tissues. Based on these analyses, the Eimeria from the Vietnamese L. calcarifer formed clades with the Eimeria detected in L. calcarifer tissues from Australia, but clustered separately from other known Eimeria and Goussia species. The Cryptosporidium detected in L. calcarifer from Vietnam clustered closest with C. parvum and C. hominis. In situ hybridization using DIG-labeled DNA probes generated from 18S PCR products on the Vietnamese L. calcarifer wax block tissues showed that this method could not be used to distinguish between Eimeria and Cryptosporidium, due to the conserved nature of the 18S locus. A previously published study on the morphology of parasite developmental stages and oocysts in the Vietnamese L. calcarifer tissues showed only an intestinal Eimeria infection. The Cryptosporidium could be present at very low levels undetectable by microscopy in intestines, or being ubiquitous, was a possible contaminant from feed or water. While molecular analysis is a very useful tool in the study of disease and identification of aetiological agents, this study reiterates the importance of demonstrating organisms in situ in tissues.
Publisher: Elsevier
Date: 2019
DOI: 10.1016/BS.APAR.2019.07.002
Abstract: A total of eight Giardia species are accepted. These include: Giardia duodenalis (syn. Giardia intestinalis and Giardia lamblia), which infects humans and animals, Giardia agilis, Giardia ardeae, Giardia psittaci, Giardia muris, Giardia microti, Giardia peramelis and G. cricetidarum, which infect non-human hosts including hibians, birds, rodents and marsupials. Giardia duodenalis is a species complex consisting of eight assemblages (A-H), with assemblages A and B the dominant assemblages in humans. Molecular studies to date on the zoonotic potential of Giardia in animals are problematic and are h ered by lack of concordance between loci. Livestock (cattle, sheep, goats and pigs) are predominantly infected with G. duodenalis assemblage E, which has recently been shown to be zoonotic, followed by assemblage A. In cats and dogs, assemblages A, B, C, D and F are commonly reported but relatively few studies have conducted molecular typing of humans and their pets and the results are contradictory with some studies support zoonotic transmission but the majority of studies suggesting separate transmission cycles. Giardia also infects a broad range of wildlife hosts and although much less well studied, host-adapted species as well as G. duodenalis assemblages (A-H) have been identified. Fish and other aquatic wildlife represent a source of infection for humans with Giardia via water contamination and/or consumption of undercooked fish and interestingly, assemblage B and A predominated in the two molecular studies conducted to date. Our current knowledge of the transmission dynamics of Giardia is still poor and the development of more discriminatory typing tools such as whole genome sequencing (WGS) of Giardia isolates is therefore essential.
Publisher: MDPI AG
Date: 19-11-2021
DOI: 10.3390/ANI11113307
Abstract: The enteric parasite, Cryptosporidium is a major cause of diarrhoeal illness in humans and animals worldwide. No effective therapeutics or vaccines are available and therefore control is dependent on understanding transmission dynamics. The development of molecular detection and typing tools has resulted in the identification of a large number of cryptic species and genotypes and facilitated our understanding of their potential for zoonotic transmission. Of the 44 recognised Cryptosporidium species and genotypes, 19 species, and four genotypes have been reported in humans with C. hominis, C. parvum, C. meleagridis, C. canis and C. felis being the most prevalent. The development of typing tools that are still lacking some zoonotic species and genotypes and more extensive molecular epidemiological studies in countries where the potential for transmission is highest are required to further our understanding of this important zoonotic pathogen. Similarly, whole-genome sequencing (WGS) and licon next-generation sequencing (NGS) are important for more accurately tracking transmission and understanding the mechanisms behind host specificity.
Publisher: Elsevier BV
Date: 02-2018
DOI: 10.1016/J.EXPPARA.2018.01.011
Abstract: Little is known about the prevalence of Giardia duodenalis in human patients in Jordan and all previous studies have used direct microscopy, which lacks sensitivity. The present study developed a novel quantitative PCR (qPCR) assay at the β-giardin (bg) locus and evaluated its use as a frontline test for the diagnosis of giardiasis in comparison with a commercially available ELISA using nested PCR and sequencing of the glutamate dehydrogenase (gdh) locus (gdh nPCR) as the gold standard. A total of 96 human faecal s les were collected from 96 patients suffering from diarrhoea from 5 regions of Jordan and were screened using the ELISA and qPCR. The analytical specificity of the bg qPCR assay revealed no cross-reactions with other genera and detected all the Giardia isolates tested. Analytical sensitivity was 1 Giardia cyst per μl of DNA extract. The overall prevalence of Giardia was 64.6%. The clinical sensitivity and specificity of the bg qPCR was 89.9% and 82.9% respectively compared to 76.5 and 68.0% for the ELISA. This study is the first to compare three different methods (ELISA, bg qPCR, nested PCR and sequencing at the gdh locus) to diagnose Jordanian patients suffering from giardiasis and to analyze their demographic data.
Publisher: Cambridge University Press (CUP)
Date: 10-07-2015
DOI: 10.1017/S0031182015000785
Abstract: Trypanosomes are blood-borne parasites that can cause severe disease in both humans and animals, yet little is known of the pathogenicity and life-cycles of trypanosomes in native Australian mammals. Trypanosoma copemani is known to be infective to a variety of Australian marsupials and has recently been shown to be potentially zoonotic as it is resistant to normal human serum. In the present study, in vivo and in vitro examination of blood and cultures from Australian marsupials was conducted using light microscopy, immunofluorescence, scanning electron microscopy and fluorescence in situ hybridization. Promastigote, sphaeromastigote and amastigote life-cycle stages were detected in vivo and in vitro . Novel trypanosome-like stages were also detected both in vivo and in vitro representing an oval stage, an extremely thin stage, an adherent stage and a tiny round stage. The tiny round and adherent stages appeared to adhere to erythrocytes causing potential haematological damage with clinical effects similar to haemolytic anaemia. The present study shows for the first time that trypomastigotes are not the only life-cycle stages circulating within the blood stream of trypanosome infected Australian native marsupials and provides insights into possible pathogenic mechanisms of this potentially zoonotic trypanosome species.
Publisher: Elsevier BV
Date: 02-2013
DOI: 10.1016/J.EXPPARA.2012.11.012
Abstract: A new species, Eimeria tiliquae n. sp. is described from a shingleback skink (Tiliqua rugosa rugosa). Sporulated oocysts (n=50) are spherical to subspherical, with colourless trilaminate oocyst wall, 0.7±0.1 (0.5-0.75) thick. Oocyst with 4 spheroidal to subspheroidal sporocysts. Oocyst length, 13.7±0.9 (12.0-16.3) oocyst width, 12.8±0.9 (11.5-15.0) oocyst length/width (L/W) ratio, 1.07±0.05 (1.0-1.2). Micropyle, oocyst residuum and polar granule absent. Sporocysts with globular sporocyst residuum and 2 sporozoites. Sporocyst length, 6.0±0.6 (5.0-7.5) sporocyst width, 5.4±0.6 (4.0-7.0) sporocyst L/W ratio, 1.11±0.11 (1.0-1.5). Stieda, parastieda and substieda bodies absent. Phylogenetic analysis of 18S rRNA sequences indicated that E. tiliquae n. sp. shared 96.4-96.5% genetic similarity to E. tropidura, its closest relative. Reptile-derived sequences were not available for the mitochondrial cytochrome oxidase gene (COI) and phylogenetic analysis at this locus placed E. tiliquae n. sp. in a clade by itself but grouping closest (92% similarity) with a novel isolate from a King's skink (Egernia kingii) from Western Australia. Based on morphological and molecular data, this isolate is a new species of coccidian parasite that to date has only been found in shingleback skinks.
Publisher: Elsevier BV
Date: 06-2013
DOI: 10.1016/J.MEEGID.2013.01.011
Abstract: In order to examine the prevalence of Cryptosporidium in wild rodents in the Philippines and understand the role wild rodents play in the transmission of this parasite to humans and livestock, 194 fecal s les from wild rats and mice from Luzon and Mindoro islands were examined. Molecular screening at the 18S and actin gene loci identified an overall prevalence of 25.8% (95%CI: 19.8, 32.5). Sequence and phylogenetic analysis of both loci identified C. parvum, C. muris, C. scrofarum, rat genotypes I-IV and a C. suis-like genotype in the rat-derived isolates and is the first report of C. suis-like and C. scrofarum in rats. Mixed infections were identified in 24% of the Cryptosporidium positive isolates. Rat genotypes II, III and IV showed high intragenotypic variation at the 18S gene locus compared to the actin locus.
Publisher: Proceedings of the National Academy of Sciences
Date: 21-10-2019
Abstract: Neospora caninum , a cyst-forming apicomplexan parasite, is a leading cause of neuromuscular diseases in dogs as well as fetal abortion in cattle worldwide. The importance of the domestic and sylvatic life cycles of Neospora , and the role of vertical transmission in the expansion and transmission of infection in cattle, is not sufficiently understood. To elucidate the population genomics of Neospora , we genotyped 50 isolates collected worldwide from a wide range of hosts using 19 linked and unlinked genetic markers. Phylogenetic analysis and genetic distance indices resolved a single genotype of N. caninum . Whole-genome sequencing of 7 isolates from 2 different continents identified high linkage disequilibrium, significant structural variation, but only limited polymorphism genome-wide, with only 5,766 biallelic single nucleotide polymorphisms (SNPs) total. Greater than half of these SNPs (∼3,000) clustered into 6 distinct haploblocks and each block possessed limited allelic ersity (with only 4 to 6 haplotypes resolved at each cluster). Importantly, the alleles at each haploblock had independently segregated across the strains sequenced, supporting a unisexual expansion model that is mosaic at 6 genomic blocks. Integrating seroprevalence data from African cattle, our data support a global selective sweep of a highly inbred livestock pathogen that originated within European dairy stock and expanded transcontinentally via unisexual mating and vertical transmission very recently, likely the result of human activities, including recurrent migration, domestication, and breed development of bovid and canid hosts within similar proximities.
Publisher: Elsevier BV
Date: 04-1996
DOI: 10.1016/0166-6851(96)02577-7
Abstract: Fritillaria species, a well-known Chinese traditional medicine for more than 2,000 years, have become rare resources due to excessive harvesting. In order to balance the economical requirement and ecological protection of
Publisher: Elsevier BV
Date: 2012
DOI: 10.1016/J.TVJL.2011.01.007
Abstract: Two hundred and forty calf faecal s les from 16 Malaysian farms were screened by PCR for Giardia spp. The overall prevalence was 12.5% and the overall farm prevalence was 68.8% (11/16 farms). The prevalence in pre-weaned and weaned calves was 16.7% and 8.3%, respectively. Sequence analysis of 25 isolates identified all as G. duodenalis assemblage E. Management factors associated with an increased risk of infection with Giardia spp. included keeping weaned calves in pens with sand floors and calf age. Keeping pre-weaned calves in pens with concrete floors and calving in single cow calving areas decreased the risk.
Publisher: Elsevier BV
Date: 07-2022
DOI: 10.1016/J.WATRES.2022.118659
Abstract: As urban communities continue to grow, demand for recreational access (including swimming) in drinking water sources have increased, yet relatively little is understood about the public health implications this poses for drinking water consumers. Preventative risk-based approaches to catchment management, informed by quantitative microbial risk assessment (QMRA), requires accurate input data to effectively model risks. A sound understanding of the knowledge gaps is also important to comprehend levels of uncertainty and help prioritise research needs. Cryptosporidium is one of the most important causes of waterborne outbreaks of gastroenteritis globally due to its resistance to chlorine. This review was undertaken by Water Research Australia to provide the most up-to-date information on current Cryptosporidium epidemiological data and underlying assumptions for exposure assessment, dose response and risk assessment for generic components of QMRA for Cryptosporidium and highlights priorities for common research. Key interim recommendations and guidelines for numerical values for relatively simple screening level QMRA modelling are provided to help support prospective studies of risks to drinking water consumers from Cryptosporidium due to body-contact recreation in source water. The review does not cover site-specific considerations, such as the levels of activity in the source water, the influence of dilution and inactivation in reservoirs, or water treatment. Although the focus is Australia, the recommendations and numerical values developed in this review, and the highlighted research priorities, are broadly applicable across all drinking source water sources that allow recreational activities.
Publisher: JSTOR
Date: 10-1997
DOI: 10.2307/3284275
Publisher: Elsevier BV
Date: 12-2015
DOI: 10.1016/J.PROTIS.2015.10.001
Abstract: Piroplasms, tick-transmitted Apicomplexa of the genera Theileria, Babesia and Cytauxzoon, are blood-borne parasites of clinical and veterinary importance. The order Piroplasmida shows a puzzling systematics characterized by multiple clades, soft polytomies and paraphyletic olyphyletic genera. In the present study, screening of platypuses (Ornithorhynchus anatinus), was performed to infer the parasite molecular phylogeny. DNA was extracted from blood, ectoparasites and tick eggs and the 18S rRNA- hsp70-genes were used for the phylogenetic reconstructions. Microscopic analyses detected pleomorphic intra-erythrocytic organisms and tetrads consistent with previous descriptions of Theileria ornithorhynchi Mackerras, 1959, but observation of possible schizonts could not be confirmed. DNA sequences obtained from blood and ticks allowed resolving the systematics of the first piroplasm infecting a monotreme host. Molecularly, T. ornithorhynchi formed a novel monophyletic group, basal to most known piroplasms' clades. The ancestral position of this clade, isolated from an ancient lineage of mammalian host appears particularly fascinating. The present paper discusses the inadequacies of the current molecular systematics for the Piroplasmida and the consequences of incomplete s ling, morphology-based classification and ambiguous microscopic identifications. Likely when the current s ling bias is rectified and more sequence data is made available, the phylogenetic position of T. ornithorhynchi will be further contextualized without ambiguity.
Publisher: Elsevier BV
Date: 08-2015
DOI: 10.1016/J.EXPPARA.2015.05.001
Abstract: Little is known of the prevalence of Cryptosporidium and Giardia in domestic cats in Western Australia and their potential role as zoonotic reservoirs for human infection. In the present study, a total of 345 faecal s les from four different sources were screened for the presence of Cryptosporidium and Giardia by PCR and genotyped by sequence analysis. Oocyst numbers and cyst numbers for Cryptosporidium and Giardia respectively were also determined using quantitative PCR assays. Cryptosporidium and Giardia were detected in 9.9% (95% CI 6.7-13.0) and 10.1% (95% CI 7.0-13.3) of cats in Western Australia respectively. Sequence analysis at the 18S rRNA locus identified five Cryptosporidium species/genotypes C. felis (n = 8), C. muris (n = 1), C. ryanae (n = 1), Cryptosporidium rat genotype III (n = 5) and a novel genotype most closely related to Cryptosporidium rat genotype III in one isolate. This is the first report of C. ryanae and Cryptosporidium rat genotype III in cats. For Giardia, assemblage F the most commonly identified species, while only 1 assemblage sequence was detected. Since most human cases of cryptosporidiosis are caused by C. parvum and C. hominis and human cases of giardiasis are caused by G. duodenalis assemblage A and B, the domestic cats in the present study are likely to be of low zoonotic risk to pet owners in Perth. Risk analyses identified that elderly cats (more than 6 years) were more prone to Cryptosporidium and Giardia infections than kittens (less than 6 months) (P = 0.009). Clinical symptoms were not associated with the prevalence of Cryptosporidium and Giardia infections in cats.
Publisher: Springer Science and Business Media LLC
Date: 12-07-2018
DOI: 10.1007/S00436-018-5989-1
Abstract: A survey was conducted on 30 Danish mink farms from April to October 2016 to determine the prevalence and species of Eimeria in Danish farmed mink. In total, 2.6% of mink faecal s les (108/4140) were positive for Eimeria vison-like oocysts by microscopy, with 24.8% (78/315) of mink being positive at least once during the study period. Morphological analysis of sporulated oocysts (n = 20) identified Eimeria vison-like oocysts measuring 21.0 × 13.8 μm with a length/width (L/W) ratio of 1.5. Phylogenetic analysis of 18S rRNA sequences (1221 bp) from three positive mink indicated that Eimeria vison-like shared the highest genetic similarity to Eimeria sp. ex Apodemus agrarius from a Striped field mouse (A. agrarius) from the Czech Republic (99.6%). Analysis of a shorter region of 18S (531 bp) revealed that the E. vison-like genotype sequences grouped in the same clade and shared 97.7% similarity with E. furonis. At the cytochrome c oxidase subunit I (COI) locus, mink-derived sequences were not available from GenBank and phylogenetic analysis placed the novel E. vison-like in a clade with E. cf. ictidea (99.4% similarity) from a black footed ferret (Mustela nigripes) from Canada.
Publisher: Oxford University Press (OUP)
Date: 05-2005
DOI: 10.1111/J.1365-2672.2005.02562.X
Abstract: Currently cryptosporidiosis represents the major public health concern of water utilities in developed nations and increasingly, new species and genotypes of Cryptosporidium are being identified in which the infectivity for humans is not clear. The complicated epidemiology of Cryptosporidium and the fact that the majority of species and genotypes of Cryptosporidium cannot be distinguished morphologically makes the assessment of public health risk difficult if oocysts are detected in the raw water supplies. The aim of this study was to use molecular tools to identify sources of Cryptosporidium from the Warragamba catchment area of Sydney, Australia. Both faecal and water s les from the catchment area were collected and screened using immunomagnetic separation (IMS) and immunofluorescence microscopy. S les that contained Cryptosporidium oocysts were genotyped using sequence and phylogenetic analysis of the 18S rDNA, and the heat-shock (HSP-70) gene. Analysis identified five Cryptosporidium species/genotypes including C. parvum (cattle genotype), C. suis, pig genotype II, the cervid genotype and a novel goat genotype. Monitoring and characterization of the sources of oocyst contamination in watersheds will aid in the development and implementation of the most appropriate watershed management policies to protect the public from the risks of waterborne Cryptosporidium. This study has shown that quantification by IMS analysis can be combined with the specificity of genotyping to provide an extremely valuable tool for assessing the human health risks from land use activities in drinking water catchments.
Publisher: Elsevier BV
Date: 12-2011
DOI: 10.1016/J.MEEGID.2011.07.021
Abstract: Fleas are important arthropod vectors for a variety of diseases in veterinary and human medicine, and bacteria belonging to the genus Bartonella are among the organisms most commonly transmitted by these ectoparasites. Recently, a number of novel Bartonella species and novel species candidates have been reported in marsupial fleas in Australia. In the present study the genetic ersity of marsupial fleas was investigated 10 species of fleas were collected from seven different marsupial and placental mammal hosts in Western Australia including woylies (Bettongia penicillata), western barred bandicoots (Perameles bougainville), mardos (Antechinus flavipes), bush rats (Rattus fuscipes), red foxes (Vulpes vulpes), feral cats (Felis catus) and rabbits (Oryctolagus cuniculus). PCR and sequence analysis of the cytochrome oxidase subunit I (COI) and the 18S rRNA genes from these fleas was performed. Concatenated phylogenetic analysis of the COI and 18S rRNA genes revealed a close genetic relationship between marsupial fleas, with Pygiopsylla hilli from woylies, Pygiopsylla tunneyi from western barred bandicoots and Acanthopsylla jordani from mardos, forming a separate cluster from fleas collected from the placental mammals in the same geographical area. The clustering of Bartonella species with their marsupial flea hosts suggests co-evolution of marsupial hosts, marsupial fleas and Bartonella species in Australia.
Publisher: Elsevier BV
Date: 2016
DOI: 10.1016/J.EXPPARA.2015.11.001
Abstract: A new species, Choleoeimeria pogonae n. sp. is described from a Western bearded dragon (Pogona minor minor) in Western Australia. Sporulated oocysts (n = 48) were cylindroidal in shape. Oocyst length, 27.0 (26.0-28.3) μm, oocyst width, 15.2 (14.0-16.5) μm, oocyst length/width ratio (L/W) 1.8 (1.6-1.9), each with 4 sporocysts (Eimeria-like) and a polar granule, but lacking a micropyle and oocyst residuum. Sporocysts are ovoidal in shape, sporocyst length, 10.0 (9.0-11.0) μm, sporocyst width 8.5 (7.0-9.5) μm, sporocyst L/W ratio, 1.2 (1.1-1.3). Stieda, substieda and parasubstieda bodies were all absent. Molecular analysis was conducted at the 18S rRNA and cytochrome c oxidase I (COI) loci. Phylogenetic analysis of 18S sequences revealed that C. pogonae n. sp. grouped together with another four Choleoeimeria spp. and exhibited 99.1%-99.4% genetic similarity. At the COI locus, C. pogonae n. sp. was in an independent clade and had the highest similarity (80.4%) to Eimeria cf. mivati from a chicken (Gallus gallus domesticus). According to the morphological and molecular data, this isolate is a new species of coccidian parasite. This study further supports the taxonomy of Choleoeimeria spp. as a new genus based on molecular phylogenetic analysis.
Publisher: Informa UK Limited
Date: 29-05-2023
Publisher: Elsevier BV
Date: 04-2015
DOI: 10.1016/J.EXPPARA.2014.08.018
Abstract: A new species, Isospora streperae n. sp., (Apicomplexa: Eimeriidae) is described from a single grey currawong bird (Strepera versicolour) (subspecies S. v. plumbea) in Western Australia. Sporulated oocysts (n = 32) are spherical to subspherical, with smooth colourless bilayered oocyst wall, 1.0 µm thick (outer layer 0⋅8 µm, inner 0.2 µm thick). Oocyst with a polar granule, an oocyst residuum and two spheroidal to subspheroidal sporocysts. Oocyst length, 23.8 (20.4-25.0) µm oocyst width, 22.5 (20.0-24.6) µm a shape index of 1.06, with Stieda, substieda bodies. Micropyle is absent. Sporocysts with compressed sporocyst residuum and four sporozoites. Sporocyst length, 14.4 (12.5-15.2) µm sporocyst width, 11.2 (10.6-14.0) µm, sporocyst L/W ratio, 1.29. Necropsy of the bird identified haemorrhaging along the ileum and jejunum, which is where Isospora oocysts were also mostly detected. Molecular analysis was conducted at three loci the 18S, 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) gene. At the 18S locus, I. streperae n. sp. exhibited 99.5% and 99.4% similarity respectively to an Isospora sp. (MS-2003) from a Southern cape sparrow (Passer melanurus melanurus) and Isospora dovati from a domestic pigeon (Columba livia domestica). At the 28S locus, I. streperae n. sp. exhibited 96.9% similarity to an Isospora sp. (MS-2003) from a grosbeak starling (Scissirostrum dubium) and 95.8% similarity with the Isospora sp. (MS-2003) from a Southern cape sparrow. At the COI locus, I. streperae n. sp. exhibited 95.0% similarity to Isospora sp. from a yellow-necked mouse (Apodemus flavicollis) from the Czech Republic. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora streperae n. sp. after its host, the grey currawong (Strepera versicolour plumbea).
Publisher: Elsevier BV
Date: 07-2019
DOI: 10.1016/J.WATRES.2019.04.041
Abstract: While some microbial eukaryotes can improve effluent quality in wastewater treatment plants (WWTPs), eukaryotic waterborne pathogens are a threat to public health. This study aimed to identify Eukarya, particularly faecal pathogens including Cryptosporidium, in different treatment stages (influent, intermediate and effluent) from four WWTPs in Western Australia (WA). Three WWTPs that utilise stabilisation ponds and one WWTP that uses activated sludge (oxidation ditch) treatment technologies were s led. Eukaryotic 18S rRNA (18S) was targeted in the wastewater s les (n = 26) for next-generation sequencing (NGS), and a mammalian-blocking primer was used to reduce the lification of mammalian DNA. Overall, bioinformatics analyses revealed 49 eukaryotic phyla in WWTP s les, and three of these phyla contained human intestinal parasites, which were primarily detected in the influent. These human intestinal parasites either had a low percent sequence composition or were not detected in the intermediate and effluent stages and included the amoebozoans Endolimax sp., Entamoeba sp. and Iodamoeba sp., the human pinworm Enterobius vermicularis (Nematoda), and Blastocystis sp. subtypes (Sarcomastigophora). Six Blastocystis subtypes and four Entamoeba species were identified by eukaryotic 18S NGS, however, Cryptosporidium sp. and Giardia sp. were not detected. Real-time polymerase chain reaction (PCR) also failed to detect Giardia, but Cryptosporidium-specific NGS detected Cryptosporidium in all WWTPs, and a total of nine species were identified, including five zoonotic pathogens. Although eukaryotic 18S NGS was able to identify some faecal pathogens, this study has demonstrated that more specific NGS approaches for pathogen detection are more sensitive and should be applied to future wastewater pathogen assessments.
Publisher: Elsevier BV
Date: 08-2014
DOI: 10.1016/J.WATRES.2014.04.023
Abstract: Reliable identification of cyanobacterial isolates has significant socio-economic implications as many bloom-forming species affect the aesthetics and safety of drinking water, through the production of taste and odour compounds or toxic metabolites. The limitations of morphological identification have promoted the application of molecular tools, and encouraged the adoption of combined (polyphasic) approaches that include both microscopy- and DNA-based analyses. In this context, the rapid expansion of available sequence data is expected to allow increasingly reliable identification of cyanobacteria, and ultimately resolve current discrepancies between the two approaches. In the present study morphological and molecular characterisations of cyanobacterial isolates (n = 39), collected from various freshwater sites in Australia, were compared. Sequences were obtained for the small ribosomal subunit RNA gene (16S rDNA) (n = 36), the DNA-dependent RNA polymerase gene (rpoC1) (n = 22), and the phycocyanin operon, with its intergenic spacer region (cpcBA-IGS) (n = 19). Phylogenetic analyses identified three cyanobacterial orders: the Chroococcales (n = 8), Oscillatoriales (n = 6), and Nostocales (n = 25). Interestingly, multiple novel genotypes were identified, with 22% of the strains (17/77) having <95% similarity to available sequences in GenBank. Morphological and molecular data were in agreement at the species level for only 26% of the isolates obtained (10/39), while agreement at the genus level was obtained for 31% (12/39). Confident identification of the remaining 44% of the strains (17/39) beyond the order level was not possible. The present study demonstrates that, despite the taxonomic revisions, and advances in molecular-, and bioinformatics-tools, the lack of reliable morphological features, culture-induced pleomorphism, and proportion of misidentified or poorly described sequences in GenBank, still represent significant factors, impeding the confident identification of cyanobacteria species.
Publisher: American Society for Microbiology
Date: 2011
DOI: 10.1128/JCM.01329-10
Abstract: Although widely used for the characterization of the transmission of intestinal Cryptosporidium spp., genotyping tools are not available for C. muris and C. andersoni , two of the most common gastric Cryptosporidium spp. infecting mammals. In this study, we screened the C. muris whole-genome sequencing data for microsatellite and minisatellite sequences. Among the 13 potential loci (6 microsatellite and 7 minisatellite loci) evaluated by PCR and DNA sequencing, 4 were eventually chosen. DNA sequence analyses of 27 C. muris and 17 C. andersoni DNA preparations showed the presence of 5 to 10 subtypes of C. muris and 1 to 4 subtypes of C. andersoni at each locus. Altogether, 11 C. muris and 7 C. andersoni multilocus sequence typing (MLST) subtypes were detected among the 16 C. muris and 12 C. andersoni specimens successfully sequenced at all four loci. In all analyses, the C. muris isolate (TS03) that originated from an East African mole rat differed significantly from other C. muris isolates, approaching the extent of genetic differences between C. muris and C. andersoni . Thus, an MLST technique was developed for the high-resolution typing of C. muris and C. andersoni . It should be useful for the characterization of the population genetics and transmission of gastric Cryptosporidium spp.
Publisher: Elsevier BV
Date: 11-2013
DOI: 10.1016/J.IJPARA.2013.06.001
Abstract: Giardia duodenalis (syn. Giardia lamblia and Giardia intestinalis) is a common intestinal parasite of humans and mammals worldwide. Assessing the zoonotic transmission of the infection requires molecular characterization as there is considerable genetic variation within G. duodenalis. To date eight major genetic groups (assemblages) have been identified, two of which (A and B) are found in both humans and animals, whereas the remaining six (C to H) are host-specific and do not infect humans. Sequence-based surveys of single loci have identified a number of genetic variants (genotypes) within assemblages A and B in animal species, some of which may have zoonotic potential. Multi-locus typing data, however, has shown that in most cases, animals do not share identical multi-locus types with humans. Furthermore, interpretation of genotyping data is complicated by the presence of multiple alleles that generate "double peaks" in sequencing files from PCR products, and by the potential exchange of genetic material among isolates, which may account for the non-concordance in the assignment of isolates to specific assemblages. Therefore, a better understanding of the genetics of this parasite is required to allow the design of more sensitive and variable subtyping tools, that in turn may help unravel the complex epidemiology of this infection.
Publisher: Royal Society of Chemistry (RSC)
Date: 2021
DOI: 10.1039/D1EW00129A
Abstract: Better chemical removal in waste stabilisation ponds was linked to the presence of a maturation pond, higher solar irradiation, and warmer temperatures.
Publisher: Elsevier BV
Date: 09-2011
DOI: 10.1016/J.PREVETMED.2011.05.016
Abstract: In this study, 96 faecal s les were collected from pregnant Merino ewes, at two broad-acre, commercial sheep farms in southern Western Australia, on two separate occasions (16 and 2 weeks prior to lambing). Following lambing, 111 (Farm A) and 124 (Farm B) female crossbred lambs (2-6 weeks old), were in idually identified using ear tags (a numbered tag and a radio-frequency tag). A total of 1155 faecal s les were collected only from these in idually identified lambs on five separate s ling occasions. All s les were screened using PCR to detect Cryptosporidium (18S rRNA and actin loci) and Giardia duodenalis (glutamate dehydrogenase and triosephosphate isomerise loci). The overall prevalences (lambs positive for a parasite on at least one of the five s lings) at Farm A and B were 81.3% and 71.4%, respectively for Cryptosporidium and similarly 67.3% and 60.5% for Giardia, respectively. Cryptosporidium and Giardia prevalences at in idual s lings ranged between 18.5 and 42.6% in lambs and were <10% in the ewes. Cryptosporidium xiaoi was the most prevalent species detected at all five s lings and was also isolated from lamb dam water on Farm B. Cryptosporidium ubiquitum was most commonly detected in younger lambs and Cryptosporidium parvum was detected in lambs at all five s lings, typically in older lambs and as part of a mixed species infection with C. xiaoi. A novel, possibly new genotype (sheep genotype I), was identified in six Cryptosporidium isolates from Farm B. Giardia duodenalis assemblage E was the most common genotype detected at all five s lings, with greater proportions of assemblage A and mixed assemblage A and E infections identified in older lambs. This longitudinal study identified high overall prevalences of Cryptosporidium and Giardia in lambs grazed extensively on pastures, while reinforcing that s ling a random selection of animals from a flock/herd on one occasion (point prevalence), underestimates the overall prevalence of these parasites in the flock/herd across an extended time period. Based on these findings, grazing lambs were identified as a low risk source of zoonotic Cryptosporidium and Giardia species/genotypes, with these protozoa detected at all five s lings in some lambs, indicating that these in iduals were either unable to clear the naturally acquired protozoan infections or were repeatedly re-infected from their environment or other flock members.
Publisher: Elsevier BV
Date: 12-2012
DOI: 10.1016/J.VETPAR.2012.06.036
Abstract: Current knowledge on the prevalence and genotypes of Cryptosporidium in fishes is still limited. This study investigated the prevalence of Cryptosporidium species in 171 ornamental fishes, belonging to 33 species, collected from 8 commercial aquariums around Perth, Western Australia. All s les were screened by nested PCR targeting the 18S rRNA locus. A total of 6 positives were identified by PCR at the 18S locus from 4 different species of fishes (red eye tetra, Moenkhausia sanctaefilomenae gold gourami, Trichogaster trichopterus neon tetra, Paracheirodon innesi goldfish, Carassius auratus auratus), giving an overall prevalence of 3.5% (6/171). Four different genotypes were identified, only one of which has been previously reported in fish piscine genotype 4 in a neon tetra isolate, a rat genotype III-like isolate in a goldfish, a novel genotype in three isolates from red eye (piscine genotype 7) which exhibited a 3.5% genetic distance from piscine genotype 1 and a piscine genotype 6-like from a gold gourami (1% genetic distance). Further biological and genetic characterisation is required to determine the relationship of these genotypes to established species and strains of Cryptosporidium.
Publisher: Cambridge University Press (CUP)
Date: 08-2003
DOI: 10.1017/S0950268803008781
Abstract: In a study to estimate the frequency of Cryptosporidium infections in Switzerland, stool s les from patients found to be positive for Cryptosporidium spp. by modified Ziehl–Neelson staining and fluorescence microscopy were used for genotyping experiments. With 9 of 12 s les, DNA extraction and subsequent genotyping was successful. All Cryptosporidium -isolates belonged to the bovine genotype. In one stool s le, two strains of Cryptosporidium were demonstrated, suggesting a mixed infection. In comparison with reference strains from calves, one of the isolates showed a full sequence identity and the other a similarity of 97·5%. The fact that only bovine genotypes were detected suggests, that cryptosporidiosis must primarily be considered as a zoonotic disease in Switzerland. This is in contrast to other countries, where the human genotype of C. parvum was shown to dominate the epidemiological situation. The results of our study are supported by the previous finding, that two of the analysed strains originated from patients who used to consume raw milk or raw cream, a known risk factor for cryptosporidiosis.
Publisher: American Society of Parasitologists
Date: 10-2008
DOI: 10.1645/GE-1508.1
Publisher: Springer Science and Business Media LLC
Date: 08-10-2021
Publisher: American Society for Microbiology
Date: 06-2000
DOI: 10.1128/AEM.66.6.2385-2391.2000
Abstract: We have characterized the nucleotide sequences of the 70-kDa heat shock protein (HSP70) genes of Cryptosporidium baileyi , C. felis , C. meleagridis , C. muris , C. serpentis , C. wrairi , and C. parvum from various animals. Results of the phylogenetic analysis revealed the presence of several genetically distinct species in the genus Cryptosporidium and eight distinct genotypes within the species C. parvum . Some of the latter may represent cryptic species. The phylogenetic tree constructed from these sequences is in agreement with our previous results based on the small-subunit rRNA genes of Cryptosporidium parasites. The Cryptosporidium species formed two major clades: isolates of C. muris and C. serpentis formed the first major group, while isolates of C. felis , C. meleagridis , C. wrairi , and eight genotypes of C. parvum formed the second major group. Sequence variations were also observed between C. muris isolates from ruminants and rodents. The HSP70 gene provides another useful locus for phylogenetic analysis of the genus Cryptosporidium .
Publisher: Environmental Health Perspectives
Date: 03-2006
DOI: 10.1289/EHP.8240
Abstract: A workshop titled "Application of Genotyping Methods to Assess Pathogen Risks from Cryptosporidium in Drinking Water Catchments" was held at the International Water Association biennial conference, Marrakech, Morocco, 23 September 2004. The workshop presented and discussed the findings of an interlaboratory trial that compared methods for genotyping Cryptosporidium oocysts isolated from feces. The primary goal of the trial and workshop was to assess the utility of current Cryptosporidium genotyping methods for determining the public health significance of oocysts isolated from feces in potable-water-supply watersheds. An expert panel of 16 watershed managers, public health practitioners, and molecular parasitologists was assembled for the workshop. A subordinate goal of the workshop was to educate watershed management and public health practitioners. An open invitation was extended to all conference delegates to attend the workshop, which drew approximately 50 interested delegates. In this report we summarize the peer consensus emerging from the workshop. Recommendations on the use of current methods by watershed managers and public health practitioners were proposed. Importantly, all the methods that were reported in the trial were mutually supporting and found to be valuable and worthy of further utility and development. Where there were choices as to which method to apply, the small-subunit ribosomal RNA gene was considered to be the optimum genetic locus to target. The single-strand conformational polymorphism method was considered potentially the most valuable for discriminating to the subtype level and where a large number of s les were to be analyzed. A research agenda for protozoan geneticists was proposed to improve the utility of methods into the future. Standardization of methods and nomenclature was promoted.
Publisher: Springer Science and Business Media LLC
Date: 20-05-2016
DOI: 10.1007/S00436-016-5132-0
Abstract: Little is known about the epidemiology of Giardia in Jordan and to date, no genotyping studies have been conducted on Giardia isolates from Jordanians. In the present study, a total of 49 microscopy-positive faecal s les from Jordanian patients suffering from giardiasis were analysed at two loci: the triose phosphate isomerase (tpi) gene and the glutamate dehydrogenase (gdh) gene. At the tpi locus, a total of 28 s les lified and assemblage A was identified in 46.4 % (13/28) s les, while assemblage B was identified in 50 % (14/28) s les and a mixed assemblage A and B was identified in one s le (3.6 %) (Table 1). At the gdh locus 48 isolates lified and of these assemblages A was identified in 43.7 % (21/48) of isolates and assemblage B in 56.3 % (27/48) of isolates. No mixed infections were detected at the gdh locus. Subtyping at the gdh locus identified sub-assemblage AII in 43.7 % (21/48) of isolates and sub-assemblages BIII and BIV in 25 % (12/48) and 31.2 % (15/48) of isolates, respectively, with more genetic ersity in AII isolates than BIII or BIV isolates. Novel sub-types within each sub-assemblage were identified suggesting unique endemicity and anthroponotic transmission of Giardia in Jordanian patients suffering from giardiasis. Further studies are required to better understand the epidemiology and transmission of Giardia in Jordan.
Publisher: Wiley
Date: 25-04-2016
DOI: 10.1111/AVJ.12428
Abstract: To develop molecular tools for the investigation of the prevalence, species and faecal shedding of Yersinia in sheep. A quantitative PCR (qPCR) targeting the β subunit of the Yersinia spp. RNA polymerase gene was developed and validated. The prevalence of pathogenic Y. enterocolitica was determined by screening for the virulent yst gene. These qPCR assays were used to determine Yersinia spp. prevalence and faecal shedding concentration from 3412 faecal s les collected from approximately 1189 lambs (100-180 lambs/flock) on eight farms across Australia. This was a longitudinal study, with sheep s led on three occasions (weaning, post-weaning and pre-slaughter). A subset of up to five positive s les from each s ling on each farm (n = 111) was sequenced. Yersinia spp. (including both pathogenic and non-pathogenic species) were identified in all flocks, with 60.7% of lambs shedding Yersinia spp. on at least one s ling occasion. Point prevalence ranged from 4% to 91% across farms and s ling occasions. Median Yersinia spp. bacterial concentration was 1.1 × 10(6) , 2.8 × 10(6) and 5.6 × 10(5) organisms/g faeces at weaning, post-weaning and pre-slaughter, respectively, across all farms. Pathogenic Y. enterocolitica was identified in all eight flocks s led, with 14.8% of lambs shedding pathogenic Y. enterocolitica on at least one s ling occasion. Yersinia spp. and pathogenic Y. enterocolitica in particular were commonly identified in a s le of Australian sheep flocks using molecular techniques. Further studies into associations between faecal shedding of pathogenic Yersinia spp. and sheep productivity or clinical disease may utilise qPCR in conjunction with other diagnostic tools.
Publisher: Wiley
Date: 1999
DOI: 10.1111/J.1751-0813.1999.TB12428.X
Abstract: Genetic and biological characterisation of 12 isolates of Cryptosporidium from pigs and comparing them with Cryptosporidium isolates from humans and cattle. Cryptosporidium isolates from pigs were compared with those obtained from human and cattle using rDNA sequence analysis. The infectivity of two of the porcine isolates was determined in neonatal mice and the clinical history of the infected pigs recorded. Pig-derived isolates of Cryptosporidium exhibited two distinct genotypes a porcine genotype and a bovine genotype, which is common to cattle and other livestock. The porcine genotype did not produce any infection in neonatal mice whereas the bovine genotype did. Two distinct genetically and biologically differing strains of Cryptosporidium appeared to be associated with acute diarrhoea in pigs. Whether Cryptosporidium was a primary or secondary pathogen is unclear but warrants further investigation. As the bovine genotype is known to infect humans, the results suggest that pigs can act as reservoirs of cryptosporidial infections for humans and other live-stock. The zoonotic potential of the pig-adapted genotype is uncertain and requires further study.
Publisher: Elsevier BV
Date: 02-2011
DOI: 10.1016/J.EXPPARA.2010.10.015
Abstract: A cross-sectional study was carried out to identify species and determine the prevalence of Cryptosporidium sp. shedding in pre-weaned and post-weaned dairy calves and to identify management factors that may be contributing to disease. A total of 240 calf faecal s les were collected from 16 farms in two districts in Johor, Malaysia, and screened by PCR. The overall Cryptosporidium prevalence was 27.1%. The prevalence of Cryptosporidium species in pre-weaned calves was 32.4% for C. parvum, 26.5% for C. bovis, followed by C. andersoni (20.6%), C. ryanae (11.8%) and mixed sp. (8.8%). The prevalence of Cryptosporidium species in post-weaned calves was 35% for C. bovis followed by C. andersoni and C. ryanae (30% each) and mixed sp. (5%). Subtyping analysis of 8 of the 11 C. parvum isolates at the gp60 locus identified five isolates as IIdA15G1, one as IIa18A3R1 and two isolates as IIa17G2R1. Management factors that increased the risk of Cryptosporidium infection included having other cattle farms close by, feeding calves with saleable milk, keeping pre-weaned calves in pens with slatted floors and keeping post-weaned calves in pens with a sand floor.
Publisher: Springer Science and Business Media LLC
Date: 04-01-2018
Publisher: Elsevier BV
Date: 08-2010
DOI: 10.1016/J.EXPPARA.2010.02.017
Abstract: Cryptosporidium is a gastrointestinal parasite that is recognised as a significant cause of non-viral diarrhea in both developing and industrialised countries. In the present study, a longitudinal analysis of 248 faecal specimens from Australian humans with gastrointestinal symptoms from 2005 to 2008 was conducted. Sequence analysis of the 18S rRNA gene locus and the 60kDa glycoprotein (gp60) gene locus revealed that 195 (78.6%) of the cases were due to infection with Cryptosporidium hominis, 49 (19.8%) with Cryptosporidium parvum and four (1.6%) with Cryptosporidium meleagridis. A total of eight gp60 subtype families were identified five C. hominis subtype families (Ib, Id, Ie, If and Ig), and two C. parvum subtype families (IIa and IId). The Id subtype family was the most common C. hominis subtype family identified in 45.7% of isolates, followed by the Ig subtype family (30.3%) and the Ib subtype family (20%). The most common C. parvum subtype was IIaA18G3R1, identified in 65.3% of isolates. The more rare zoonotic IId A15G1 subtype was identified in one isolate. Statistical analysis showed that the Id subtype was associated with abdominal pain (p<0.05) and that in sporadic cryptosporidiosis, children aged 5 and below were 1.91 times and 1.88 times more likely to be infected with subtype Id (RR 1.91 95% CI, 1.7-2.89 p<0.05) and Ig (RR 1.88 95% CI, 1.10-3.24 p<0.05) compared to children aged 5 and above. A subset of isolates were also analysed at the variable CP47 and MSC6-7 gene loci. Findings from this study suggest that anthroponotic transmission of Cryptosporidium plays a major role in the epidemiology of cryptosporidiosis in Western Australian humans.
Publisher: Elsevier BV
Date: 08-2017
DOI: 10.1016/J.TTBDIS.2017.05.009
Abstract: Anaplasma and Ehrlichia spp. are tick-borne pathogens that can cause severe disease in domestic animals, and several species are responsible for emerging zoonoses in the northern hemisphere. Until recently, the only members of these genera reported in Australia (A. marginale, A. centrale, and A. platys) were introduced from other continents, through the importation of domestic animals and their associated ticks. However, unique Anaplasma and Ehrlichia 16S rRNA gene sequences were recently detected for the first time in native Australian ticks, particularly in Amblyomma triguttatum subsp. ticks from southwest Western Australia (WA). We used molecular techniques to survey Am. triguttatum subsp. ticks from four allopatric populations in southern and western Australia for Anaplasma and Ehrlichia species, and described here the phylogeny of these novel organisms. An A. bovis variant (genotype Y11) was detected in ticks from two study sites Yanchep National Park (12/280, 4.3%) and Barrow Island (1/69, 1.4%). Phylogenetic analysis of 16S rRNA and groEL gene sequences concluded that A. bovis genotype Y11 is a unique genetic variant, distinct from other A. bovis isolates worldwide. Additionally, a novel Ehrlichia species was detected in Am. triguttatum subsp. from three of the four study sites Yanchep National Park (18/280, 6.4%), Bungendore Park (8/46, 17.4%), and Innes National Park (9/214, 4.2%), but not from Barrow Island. Phylogenetic analysis of 16S, groEL, gltA, and map1 gene sequences revealed that this Ehrlichia sp. is most closely related to, but clearly distinct from, E. ruminantium and Ehrlichia sp. Panola Mountain. We propose to designate this new species 'Candidatus Ehrlichia occidentalis'. Anaplasma bovis genotype Y11 and 'Candidatus E. occidentalis' are the first Anaplasma and Ehrlichia species to be recorded in native Australian ticks.
Publisher: Elsevier BV
Date: 07-2008
DOI: 10.1016/J.EXPPARA.2008.04.009
Abstract: A total of 289 pig faecal s les were collected from pre-weaned and post-weaned piglets and sows from 1 indoor and 3 outdoor piggeries located in the south-west region of Western Australia and screened at the 18S rRNA locus for the presence of Cryptosporidium. An overall prevalence of 22.1% (64/289) was identified. Cryptosporidium was more prevalent in post-weaned animals (p<0.05) 32.7% (51/156) versus 10.6% (13/123) for pre-weaned animals. Sequence analysis identified Cryptosporidium suis in all pre-weaned isolates genotyped (7/13). In post-weaned pigs that were genotyped (n=38), the non-zoonotic Cryptosporidium species, pig genotype II was identified in 32 s les and C. suis in 6 s les. The zoonotic species Cryptosporidium parvum was not detected, suggesting that domestic pigs do not pose a significant public health risk. Pig genotype II was significantly (p<0.05) associated with 'normal' stools, indicating an asymptomatic nature in the porcine host.
Publisher: Elsevier BV
Date: 08-2010
DOI: 10.1016/J.EXPPARA.2010.02.012
Abstract: Molecular typing at the 18S rRNA and Gp60 loci was conducted on Cryptosporidium-positive stool s les from cases collected during 2007 Western Australian and South Australian outbreaks of cryptosporidiosis. Analysis of 48 Western Australian s les identified that all isolates were C. hominis and were from five different Gp60C. hominis subtype families. The IbA10G2 subtype was most common across all age groups (37/48). In South Australia, analysis of 24 outbreak s les, identified 21 C. hominis isolates, two C. parvum isolates and one s le with both C. hominis and C. parvum. All C. hominis isolates were identified as the IbA10G2 subtype.
Publisher: Elsevier BV
Date: 08-2016
DOI: 10.1016/J.PARINT.2016.03.004
Abstract: The present study describes the first report of Trypanosoma vegrandis in koalas using morphology and sequence analysis of the 18S rRNA gene. The prevalence of T. vegrandis in koalas was 13.6% (6/44). It is likely that the small size of T. vegrandis (<10μm in length), coupled with the difficulties in lifying DNA of this parasite in mixed infections using trypanosome generic primers, are the reason why this organism has not been identified in koalas until now. This study highlights the importance of further research comprising a larger s le size to determine the prevalence of T. vegrandis in koalas as well as its potential impacts upon this marsupial species' health.
Publisher: Elsevier BV
Date: 11-2013
DOI: 10.1016/J.VETPAR.2013.08.031
Abstract: There is still limited information on the distribution of Cryptosporidium species and genotypes in fish. The present study investigated the prevalence of Cryptosporidium species in cultured freshwater (n=132), wild freshwater (n=206) and wild marine (n=276) fish in Papua New Guinea (PNG) by PCR screening at the 18S rRNA locus. A total of seven fish (2 cultured freshwater, 1 wild freshwater and 4 wild marine fish) were identified as positive for Cryptosporidium. Specifically, Cryptosporidium was found in four different host species (Nile tilapia, Oreochromis niloticus silver barb, Puntius gonionotus mackerel scad, Decapterus maracellus and oblong silver biddy, Gerres oblongus), giving an overall prevalence of 1.14% (95% CI: 0.3-2%, n=7/614). Of the seven positive isolates, five were identified as C. parvum and two were a novel piscine genotype, which we have named piscine genotype 8. Piscine genotype 8 was identified in two marine oblong silver biddies and exhibited 4.3% genetic distance from piscine genotype 3 at the 18S locus. Further subtyping of C. parvum isolates at the 60 kDa glycoprotein (gp60) locus identified 3 C. parvum subtypes (IIaA14G2R1, IIaA15G2R1 and IIaA19G4R1) all of which are zoonotic and a C. hominis subtype (IdA15G1). The zoonotic Cryptosporidium were identified in fish s les from all three groups cultured and wild freshwater and wild marine fish. Detection of Cryptosporidium among aquaculture fingerlings warrants further research to gain a better understanding of the epidemiology of Cryptosporidium infection in cultured fish. The identification of zoonotic Cryptosporidium genotypes in fish from PNG has important public health implications and should be investigated further.
Publisher: Elsevier BV
Date: 2011
DOI: 10.1016/J.VETPAR.2010.10.003
Abstract: To date Cryptosporidium muris has been identified by microscopy and genotyping in cats in two studies. We report morphological and genetic evidence of a mixed C. muris and C. felis infection in a cat and provide the first histological, immunohistochemical, in situ hybridisation and genetic confirmation of a C. muris infection in the stomach of a cat. The cat suffered persistent diarrhoea after the initial consultation, which remained unresolved, despite several medical interventions. Further studies are required to determine the range, prevalence and clinical impact of Cryptosporidium species infecting cats.
Publisher: Elsevier BV
Date: 06-2012
DOI: 10.1016/J.VETPAR.2011.12.010
Abstract: Trypanosoma evansi, a blood-borne protozoan parasite with an extensive geographical range is the causative agent of the livestock disease known as surra. A total of 140 out of 179 T. evansi isolates collected between 2006 and 2007 from 44 villages (comprising of 16 reported surra outbreaks) in 3 provinces (Agusan del Sur (ADS), Surigao del Sur (SDS) and Agusan del Norte (ADN)) in Mindanao, Philippines were each successfully genotyped using a suite of 7 polymorphic microsatellites. The study identified 16 multi locus genotypes (MLG) within the T. evansi isolates and evidence of the spread of surra outbreaks from one village to another, most likely due to the movement of infected animals. Genotyping provided evidence of population sub-structuring with 3 populations (I, II and III (only 1 isolate)) identified. The most abundant population was II, which was the predominant population in ADS and SDS (p=0.022). In addition, buffalo mortality was statistically higher in outbreak areas associated with isolates from population I (13.6%) than with isolates from population II (6.9%) (p=0.047). The present study has highlighted the utility of microsatellite loci to improve understanding of the epidemiology of T. evansi and in tracking surra outbreaks.
Publisher: Wiley
Date: 28-10-2007
DOI: 10.1111/J.1751-0813.2007.00220.X
Abstract: This study reports on the epidemiology of Babesia gibsoni in American Pit Bull Terriers living in a region of western Victoria in southern Australia. Both American Pit Bull Terriers (n = 100) and other dog breeds (n = 51) were screened for B gibsoni using immunofluorescent antibody testing (IFAT) and/or polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). A questionnaire was also completed by each dog owner, ascertaining the husbandry and habits of the dogs s led. Fourteen dogs were positive for B gibsoni using IFAT and/or PCR-RFLP and all were American Pit Bull Terriers. Dogs that were male and/or had been bitten by or were biters of other American Pit Bull Terriers were more likely to be B gibsoni positive, thus suggesting that blood-to-blood transmission contributes to the spread of this disease between dogs.
Publisher: Elsevier BV
Date: 05-2014
DOI: 10.1016/J.EXPPARA.2014.02.011
Abstract: A new species, Isospora anthochaerae n. sp. is described from a Red wattlebird (Anthochaera carunculata). Sporulated oocysts (n=37) are subspherical, with smooth colourless to pale brown bilayered oocyst wall, 0.8 μm thick (outer layer 0·6 μm, inner 0.2 μm thick). Oocyst with 2 spheroidal to subspheroidal sporocysts. Oocyst length, 23.4 μm (20.0-26.0) oocyst width, 20.7 μm (19.0-22.0) oocyst length/width (L/W) ratio, 1.1. Micropyle, oocyst residuum and polar granule are absent. Sporocysts with compact sporocyst residuum and 4 sporozoites. Sporocyst length, 14.5 μm sporocyst width, 10.1 μm sporocyst L/W ratio, 1.4. Molecular analysis was conducted at four loci the ribosomal internal transcribed spacer (ITS), the 18S and 28S ribosomal RNA and the mitochondrial cytochrome oxidase gene (COI). At the COI locus, I. anthochaerae n. sp. exhibited 98.5% similarity to Isospora lesouefi from a Regent honeyeater (Xanthomyza phrygia) and 98% similarity with an Isospora sp. (iSAT5) from a blackcap (Sylvia atricapilla). Based on morphological and molecular data, this isolate is a new species of coccidian parasite that to date has only been found in Red wattlebirds.
Publisher: Elsevier BV
Date: 12-2014
DOI: 10.1016/J.IJPARA.2014.08.004
Abstract: Clinical microbiology laboratories rely on quantitative PCR for its speed, sensitivity, specificity and ease-of-use. However, quantitative PCR quantitation requires the use of a standard curve or normalisation to reference genes. Droplet digital PCR provides absolute quantitation without the need for calibration curves. A comparison between droplet digital PCR and quantitative PCR-based analyses was conducted for the enteric parasite Cryptosporidium, which is an important cause of gastritis in both humans and animals. Two loci were analysed (18S rRNA and actin) using a range of Cryptosporidium DNA templates, including recombinant plasmids, purified haemocytometer-counted oocysts, commercial flow cytometry-counted oocysts and faecal DNA s les from sheep, cattle and humans. Each method was evaluated for linearity, precision, limit of detection and cost. Across the same range of detection, both methods showed a high degree of linearity and positive correlation for standards (R(2)⩾0.999) and faecal s les (R(2)⩾0.9750). The precision of droplet digital PCR, as measured by mean Relative Standard Deviation (RSD %), was consistently better compared with quantitative PCR, particularly for the 18S rRNA locus, but was poorer as DNA concentration decreased. The quantitative detection of quantitative PCR was unaffected by DNA concentration, but droplet digital PCR quantitative PCR was less affected by the presence of inhibitors, compared with quantitative PCR. For most templates analysed including Cryptosporidium-positive faecal DNA, the template copy numbers, as determined by droplet digital PCR, were consistently lower than by quantitative PCR. However, the quantitations obtained by quantitative PCR are dependent on the accuracy of the standard curve and when the quantitative PCR data were corrected for pipetting and DNA losses (as determined by droplet digital PCR), then the sensitivity of both methods was comparable. A cost analysis based on 96 s les revealed that the overall cost (consumables and labour) of droplet digital PCR was two times higher than quantitative PCR. Using droplet digital PCR to precisely quantify standard dilutions used for high-throughput and cost-effective lifications by quantitative PCR would be one way to combine the advantages of the two technologies.
Publisher: Springer Science and Business Media LLC
Date: 18-01-2021
Publisher: Elsevier BV
Date: 04-2008
DOI: 10.1016/J.EXPPARA.2007.10.015
Abstract: The ITS1 sequences for C. suis, C. belli, C. rivolta, C. felis, and C. ohioensis-like oocysts were determined and a diagnostic PCR-RFLP assay specific for Cystosisopora species was developed. Phylogenetic analysis of ITS1 sequences of Cystosisopora species along with ITS1 sequences for Toxoplasma, Neospora, Sarcocystis and Eimeria spp. using distance, minimum evolution and parsimony-based methods confirmed previous studies, which suggested that the genus Cystoisospora does not belong to the family Eimeriidae, but should be classified together with the cyst-forming coccidia in the family Sarcocystidae.
Publisher: Elsevier BV
Date: 04-2008
DOI: 10.1016/J.EXPPARA.2007.10.014
Abstract: Screening of 445 animal faecal s les in irrigation catchments in Western Australia (WA) was conducted to identify the prevalence of Cryptosporidium and Giardia species. Of the s les positive for Giardia duodenalis, 30.7% (12/36) were the zoonotic Assemblage A, while approximately 13% (4/30) of Cryptosporidium positives were zoonotic. This is the first finding of Giardia Assemblage A in native marsupials and birds and indicates that marsupials and possibly birds may potentially be a reservoir of zoonotic Giardia.
Publisher: Elsevier BV
Date: 12-2014
DOI: 10.1016/J.EXPPARA.2014.10.010
Abstract: An Eimeria species is described from a dusky moorhen (Gallinula tenebrosa). Sporulated oocysts (n = 40) are ovoid, with a pitted single-layered oocyst wall in young oocysts and a relatively smooth wall in the mature oocysts. Oocyst wall was 1.0 µm thick, oocysts measured 17.3 × 13.3 (16.3-17.9 × 12.7-13.9) µm, oocyst length/width (L/W) ratio, 1.3. Oocyst residuum was absent. A large polar granule was always observed in the centre of the micropyle and many small polar granules were observed when the focus was on the wall. Sporocysts are elongate-ovoid, 8.4 × 5.1 (8.0-8.9 × 4.9-5.5) µm, sporocyst L/W ratio, 1.6 (1.5-1.8), sporocyst residuum was present, composed of numerous granules in a spherical or ovoid mass. Each sporocyst contained 2 elongate sporozoites, 7.7 × 2.6 (7-10 × 2.2-3) µm. A spherical-ellipsoid posterior refractile body was found in the sporozoites. A nucleus is located immediately anterior to the posterior refractile body. When the oocyst measurements and features were compared with valid Eimeria species from hosts in the Rallidae family, this Eimeria species was identified as E. paludosa. This is the first report of E. paludosa in Australia and the dusky moorhen (Gallinula tenebrosa) in a new host for this species. Molecular analysis was conducted at three loci the 18S and 28S ribosomal RNA genes and the mitochondrial cytochrome oxidase gene (COI). At the 18 S locus, E. paludosa shared 97.3% genetic similarity with Eimeria gruis (GenBank accession number: AB544336). It also shared 99.2% genetic similarity with Eimeria crecis (GenBank accession numbers: HE653904 and HE653905) and 98.5% similarity with Eimeria nenei (GenBank accession numbers: HE653906), both of which were identified from a corncrake (Crex crex) in the United Kingdom. At the 28S locus, E. paludosa shared 91.4% similarity with E. papillata from a chicken (Gallus gallus) in the USA. At COI locus, E. paludosa was in a clade by itself and shared 87.2% similarity with E. irresidua, from a European rabbit (Oryctolagus cuniculus) from the Czech Republic. This is the first molecular characterization of E. paludosa.
Publisher: Elsevier BV
Date: 08-2012
DOI: 10.1016/J.EXPPARA.2012.05.009
Abstract: As part of a broader investigation into the potential role of black rats (Rattus rattus) as disease vectors into native small mammal populations of northern Australia, blood and faecal s les from wild black rats were screened by molecular methods, for piroplasms (Babesia and Theileria), trypanosomes and the enteric parasite Cryptosporidium. While piroplasms and trypanosomes were not detected in the blood of these animals, the overall prevalence of Cryptosporidium 18S rDNA in faecal s les was 8.2% (7/85). Co-occurrence of multiple genotypes was observed in 57.1% of the infected in iduals (4/7) cloning and re-sequencing resulted in 14 sequences which broadly grouped with Cryptosporidium sp. rat-genotypes II and III. A novel rat-derived Cryptosporidium sp. genotype at the actin locus was also obtained from five animals. The relatively low infection rate detected, and the epidemiological data on cryptosporidiosis, do not conclusively support a current threat to native Australian mammals from black rats carrying Cryptosporidium. However, this observation is based on s ling limited isolates, in limited regions. Further studies, also including s ling of native mammals, are required on larger s le sizes and from wider geographic areas, to determine the significance of these findings, including the public health importance of Cryptosporidium spp. from rodents.
Publisher: Elsevier BV
Date: 03-2010
DOI: 10.1016/J.IJPARA.2009.08.003
Abstract: Giardia duodenalis is a widespread parasite of mammalian species, including humans. Fecal s les from sporadic human clinical cases of giardiasis in Western Australia were analysed at two loci 18S rRNA and glutamate dehydrogenase (gdh), and G. duodenalis assemblage B isolates were identified in 75% of isolates. Sequence analyses of 124 isolates at the 18S rRNA locus identified 93 isolates as assemblage B and 31 as assemblage A. Analyses of 109 isolates at the gdh locus identified 44 as B3, 38 as B4 and 27 were A2. Infection with Giardia was highest amongst children 56% of infections in this age group. The majority of the isolates were from rural areas (91/124) compared with urban areas (33/124). The assemblage A isolates were completely homogenous genetically at the gdh locus, while assemblage B isolates showed variability at the nucleotide but not at the amino acid level at this locus. Some of the assemblage B3 and B4 subtypes identified in humans were previously identified in marsupials in Australia and in a fox, indicating potential zoonotic transmission.
Publisher: Elsevier
Date: 2003
Publisher: Elsevier BV
Date: 11-2016
DOI: 10.1016/J.WATRES.2016.09.013
Abstract: Parasites of the genus Cryptosporidium are a major cause of diarrhoea and ill-health in humans and animals and are frequent causes of waterborne outbreaks. Until recently, it was thought that Cryptosporidium was an obligate intracellular parasite that only replicated within a suitable host, and that faecally shed oocysts could survive in the environment but could not multiply. In light of extensive biological and molecular data, including the ability of Cryptosporidium to complete its life cycle in the absence of a host and the production of novel extracellular stages, Cryptosporidium has been formally transferred from the Coccidia, to a new subclass, Cryptogregaria, with gregarine parasites. In this review, we discuss the close relationship between Cryptosporidium and gregarines and discuss the implications for the water industry.
Publisher: Elsevier BV
Date: 11-2000
DOI: 10.1016/S0020-7519(00)00135-1
Abstract: There are 10 valid species of Cryptosporidium and perhaps other cryptic species hidden under the umbrella of Cryptosporidium parvum. The oocyst stage is of primary importance for the dispersal, survival, and infectivity of the parasite and is of major importance for detection and identification. Because most oocysts measure 4-6 microm, appear nearly spherical, and have obscure internal structures, there are few or no morphometric features to differentiate species and in vitro cultivation does not provide differential data as for bacteria. Consequently, we rely on a combination of data from three tools: morphometrics, molecular techniques, and host specificity. Of 152 species of mammals reported to be infected with C. parvum or an indistinguishable organism, very few oocysts have ever been examined using more than one of these tools. This paper reviews the valid species of Cryptosporidium, their hosts and morphometrics the reported hosts for the human pathogen, C. parvum the mechanisms of transmission the drinking water, recreational water, and food-borne outbreaks resulting from infection with C. parvum and the microscopic, immunological, and molecular methods used to detect and identify species and genotypes.
Publisher: Elsevier BV
Date: 12-2018
DOI: 10.1016/J.MEEGID.2018.09.013
Abstract: Borrelia are tick-borne bacteria that in humans are the aetiological agents of Lyme disease and relapsing fever. Here we present the first genomes of B. turcica and B. tachyglossi, members of a recently described and rapidly expanding Borrelia clade associated with reptile (B. turcica) or echidna (B. tachyglossi) hosts, transmitted by hard ticks, and of unknown pathogenicity. Borrelia tachyglossi and B. turcica genomes are similar to those of relapsing fever Borrelia species, containing a linear ~ 900 kb chromosome, a single long (> 70 kb) linear plasmid, and numerous short (< 40 kb) linear and circular plasmids, as well as a suite of housekeeping and macronutrient biosynthesis genes which are not found in Lyme disease Borrelia. Additionally, both B. tachyglossi and B. turcica contain paralogous vsp and vlp proteins homologous to those used in the multiphasic antigen-switching system used by relapsing fever Borrelia to evade vertebrate immune responses, although their number was greatly reduced compared to human-infectious species. However, B. tachyglossi and B. turcica chromosomes also contain numerous genes orthologous to Lyme disease Borrelia-specific genes, demonstrating a unique evolutionary, and potentially phenotypic link between these groups. Borrelia tachyglossi and B. turcica genomes also have unique genetic features, including degraded and deleted tRNA modification genes, and an expanded range of macronutrient salvage and biosynthesis genes compared to relapsing fever and Lyme disease Borrelia. These genomes and genomic comparisons provide an insight into the biology and evolutionary origin of these Borrelia, and provide a valuable resource for future work.
Publisher: Wiley
Date: 05-2009
DOI: 10.1111/J.1550-7408.2009.00398.X
Abstract: The morphology and genetic characterisation of a new species of piroplasm identified in the blood of the Gilbert's potoroo (Potorous gilbertii) from the Two Peoples Bay Nature Reserve near Albany, Western Australia, is described from blood and tissue s les from 16 Gilbert's potoroos. Microscopy of blood showed these parasites are highly pleomorphic with a mean length of 1.8 mum and mean width of 0.85 mum. Phylogenetic analysis of 18S rRNA sequence data identified the piroplasm as a new species of Theileria that is closely related to other Australian marsupial piroplasm species. Based on biological and molecular data, it is proposed that the parasite from Gilbert's potoroo be given the name Theileria gilberti n. sp.
Publisher: Elsevier BV
Date: 06-2019
DOI: 10.1016/J.SCITOTENV.2019.03.278
Abstract: Recycled wastewater can carry human-infectious microbial pathogens and therefore wastewater treatment strategies must effectively eliminate pathogens before recycled wastewater is used to supplement drinking and agricultural water supplies. This study characterised the bacterial composition of four wastewater treatment plants (WWTPs) (three waste stabilisation ponds and one oxidation ditch WWTP using activated sludge treatment) in Western Australia. The hypervariable region 4 (V4) of the bacterial 16S rRNA (16S) gene was sequenced using next-generation sequencing (NGS) on the Illumina MiSeq platform. Sequences were pre-processed in USEARCH v10.0 and denoised into zero-radius taxonomic units (ZOTUs) with UNOISE3. Taxonomy was assigned to the ZOTUs using QIIME 2 and the Greengenes database and cross-checked with the NCBI nr/nt database. Bacterial composition of all WWTPs and treatment stages (influent, intermediate and effluent) were dominated by Proteobacteria (29.0-87.4%), particularly Betaproteobacteria (9.0-53.5%) and Gammaproteobacteria (8.6-34.6%). Nitrifying bacteria (Nitrospira spp.) were found only in the intermediate and effluent of the oxidation ditch WWTP, and denitrifying and floc-forming bacteria were detected in all WWTPs, particularly from the families Comamonadaceae and Rhodocyclales. Twelve pathogens were assigned taxonomy by the Greengenes database, but comparison of sequences from genera and families known to contain pathogens to the NCBI nr/nt database showed that only three pathogens (Arcobacter venerupis, Laribacter hongkongensis and Neisseria canis) could be identified in the dataset at the V4 region. Importantly, Enterobacteriaceae genera could not be differentiated. Family level taxa assigned by Greengenes database agreed with NCBI nr/nt in most cases, however, BLAST analyses revealed erroneous taxa in Greengenes database. This study highlights the importance of validating taxonomy of NGS sequences with databases such as NCBI nr/nt, and recommends including the V3 region of 16S in future short licon NGS studies that aim to identify bacterial enteric pathogens, as this will improve taxonomic resolution of most, but not all, Enterobacteriaceae species.
Publisher: Elsevier BV
Date: 2021
Publisher: Elsevier BV
Date: 11-2017
DOI: 10.1016/J.MEEGID.2017.08.033
Abstract: Cryptosporidiosis is a protozoan parasitic disease which affects human and animals worldwide. In adult immunocompetent in iduals, cryptosporidiosis usually results in acute and self-limited diarrhoea however, it can cause life threatening diarrhoea in children and immunocompromised in iduals. In the present study, we compared the prevalence of Cryptosporidium species and gp60 subtypes amongst paediatric oncology patients with diarrhoea (n=160) from King Hussein Medical Centre for Cancer in Jordan, and non-oncology paediatric patients with diarrhoea (n=137) from Al-Mafraq paediatric hospital. Microscopy results using modified acid fast staining identified a significantly (p≤0.05) higher prevalence of Cryptosporidium in paediatric oncology patients with diarrhoea (14.4% - 23/160), compared to non-oncology paediatric patients with diarrhoea only (5.1% - 7/137). With the exception of one s le, all microscopy-positive s les (n=29) and an additional 3/30 microscopy-negative controls were typed to species and subtype level at the 18S and gp60 loci, respectively. All Cryptosporidium positives were typed as C. parvum. Of the 22 typed Cryptosporidium positives from the paediatric oncology patients, 21 were subtyped as IIaA17G2R1 and one as IIaA16G2R1 C. parvum subtypes. The 7 typed positives from the paediatric patients from Al-Mafraq hospital were subtyped as IIaA17G2R1 (n=5) and IIaA16G2R1 (n=2). The 3 additional positives from the 30 microscopy negative control s les were subtyped as IIaA17G2R1. The high prevalence of the IIaA17G2R1 subtype, particularly amongst oncology patients, suggests that an outbreak of cryptosporidiosis may have been occurring in oncology patients during the collection period (April to December, 2016). New therapies for cryptosporidiosis in immunocompromised patients are urgently required.
Publisher: Elsevier BV
Date: 04-2009
DOI: 10.1016/J.VETPAR.2008.12.021
Abstract: A total of 477 faecal s les from pre-weaned sheep from 5 different farms in the south west of Western Australia were screened for the presence of Cryptosporidium and Giardia using PCR. There were substantial differences in prevalence between the farms and overall prevalence was 24.5% and 11.1%, respectively for Cryptosporidium and Giardia. At the 18S locus, 66 Cryptosporidium positives were identified, the majority of which were C. bovis (n=52), followed by the cervid genotype (n=10) and C. parvum (n=2). At a second diagnostic locus, using C. parvum and C. hominis-specific qPCR primers, 63 C. parvum positives were identified, some of which were co-infections with C. bovis. The C. parvum/C. hominis qPCR was more sensitive than the nested 18S PCR at detecting C. parvum. This may be due to the low numbers of oocysts present, as quantitation data indicated that all the C. parvum detected were present in low numbers (1-10 oocysts). It may also be that using C. parvum-specific primers is necessary to determine the true prevalence of C. parvum. Amongst Giardia positive isolates, G. duodenalis genotype E (livestock) was the most prevalent (36/53), with G. duodenalis genotype A detected in five positive isolates. There were also 11 mixed A and E infections detected. The findings of the present study indicate that pre-weaned lambs are not an important source of zoonotic Giardia genotypes in Australia but may be an important source of zoonotic Cryptosporidium.
Publisher: Elsevier BV
Date: 04-2006
DOI: 10.1016/J.VETPAR.2006.01.018
Abstract: Neospora caninum was isolated and established in vitro from the skin lesion of a naturally infected dog. The identity of the parasite was evaluated by immunofluorescent antibody test (IFAT), microscopy, Western blotting and polymerase chain reaction (PCR). N. caninum DNA was detected in the whole blood, serum, skin lesion, rectal scrapings and faeces of the infected dog utilising a nested PCR targeting the Nc-5 gene of N. caninum. Antigenic and genetic characterisation of the isolate, designated WA-K9, at a number of loci including the Nc-5 gene, heat shock protein 70 (HSP-70) gene, alpha-tubulin and beta-tubulin genes revealed no variation between this isolate and two N. caninum isolates from different geographic areas. Clinical aspects of this case, which included cutaneous and neurological disease, are also discussed.
Publisher: IWA Publishing
Date: 10-11-2016
DOI: 10.2166/WH.2016.160
Abstract: Cryptosporidium is the leading cause of swimming pool outbreaks of gastroenteritis. Transmission occurs through the ingestion of oocysts that are passed in the faeces of an infected person or animal when an accidental faecal release event occurs. Cryptosporidium parasites present specific challenges for infection control as oocysts are highly resistant to chlorine levels used for pool disinfection, infected in iduals can shed large numbers of oocysts, there is a long incubation period and shedding of oocysts occurs even after symptom resolution. The purposes of this review are to identify key barriers to limiting swimming pool-associated outbreaks of cryptosporidiosis and to outline needs for research and collaboration to advance co-ordinated management practices. We reviewed swimming pool-associated cryptosporidiosis outbreaks, disinfection teachniques, current regulations and the role of staff and patrons. Key barriers to limiting swimming pool-associated outbreaks of cryptosporidiosis are a lack of uniform national and international standards, poor adherence and understanding of regulations governing staff and patron behaviour, and low levels of public knowledge and awareness.
Publisher: Elsevier BV
Date: 05-2012
DOI: 10.1016/J.EXPPARA.2012.02.021
Abstract: Piroplasms, which include the genera Theileria and Babesia, are blood-borne parasites transmitted mainly by tick vectors. Relatively little is known about their prevalence and clinical impact in Australian marsupials. In the present study the occurrence and molecular phylogeny of these parasites were studied in both wild and captive marsupials from Western Australia (WA) and Queensland (QLD). Blood s les were screened by microscopy and molecular methods, using PCR and DNA sequencing of the 18S ribosomal RNA gene (18S rDNA). Overall, 7.1% of the blood s les (8/113) were positive for piroplasm 18S rDNA. Theileria and Babesia rDNA was detected in 0.9% (1/113) and 6.2% (7/113) of the animals, respectively. The single Theileria positive was identified in one of three boodies (Bettongia lesueur) screened from a wildlife rehabilitation centre in WA, while all seven Babesia positives were detected in WA in wild captured woylies (Bettongia penicillata ogilbyi). Small intraerythrocytic inclusions were observed in blood films made from six of these in iduals. This is the first report of a Babesia sp. in woylies, and Theileria sp. in boodies. Phylogenetic analysis indicated that the woylie-derived Babesia was genetically distinct and most closely related to Babesia occultans, the causative agent of a benign form of cattle babesiosis (genetic similarity 98.4%). The Theileria identified was most closely related to the marsupial-derived species Theileria penicillata from the woylie, Theileria brachyuri from the quokka (Setonix brachyurus), and Theileria sp. from the long-nosed potoroo (Potorous tridactylus).
Publisher: American Society of Parasitologists
Date: 04-2003
Publisher: American Society for Microbiology
Date: 09-2005
DOI: 10.1128/AEM.71.9.4992-4997.2005
Abstract: Little is known of the prevalence of Cryptosporidium and Giardia parasites in sheep and the genotypes that they harbor, although potentially sheep may contribute significantly to contamination of watersheds. In the present study, conducted in Western Australia, a total of 1,647 sheep fecal s les were screened for the presence of Cryptosporidium and Giardia spp. using microscopy, and a subset ( n = 500) were screened by PCR and genotyped. Analysis revealed that although both parasites were detected in a high proportion of s les by PCR (44% and 26% for Giardia and Cryptosporidium spp., respectively), with the exception of one Cryptosporidium hominis isolate, the majority of isolates genotyped are not commonly found in humans. These results suggest that the public health risk of sheep-derived Cryptosporidium and Giardia spp. in catchment areas and effluent may be overestimated and warrant further investigation.
Publisher: Wiley
Date: 23-08-2017
Publisher: Elsevier BV
Date: 04-2021
Publisher: Elsevier BV
Date: 08-2008
DOI: 10.1016/J.MOLBIOPARA.2008.04.006
Abstract: Giardia duodenalis is a widespread parasite of mammalian species, including humans. Due to its invariant morphology, investigation on aspects such as host specificity and transmission patterns requires a direct genetic characterization of cysts/trophozoites from host s les. A number of molecular assays have been developed to help unravel the complex epidemiology of this infection. A coherent picture has emerged from those studies, indicating the existence of seven genetic groups (or assemblages), two of which (A and B) are found in both humans and animals, whereas the remaining five (C-G) are host-specific. Sequence-based surveys have identified a number of genotypes within assemblages A and B in animal species, some of which may have zoonotic potential. Recently, however, molecular approaches have been complicated by the recognition of intra-isolate sequence heterogeneity (i.e., "mixed templates", that affects identification of subtypes within each assemblage), and by the unreliable assignment of isolates to G. duodenalis assemblages generated by different genetic markers. This raises concerns about previous interpretation of genotyping data, especially when single genetic markers have been used. The mechanisms that may be responsible for these findings, including allelic sequence heterozygosity and meiotic recombination, are discussed.
Publisher: Cold Spring Harbor Laboratory
Date: 17-10-2019
DOI: 10.1101/807131
Abstract: Ticks (Acari: Ixodida) transmit a greater variety of pathogens than any other blood-feeding group of arthropods. While numerous microbes have been identified inhabiting Australian Ixodidae, some of which are related to globally important tick-borne pathogens, little is known about the bacterial communities within ticks collected from Australian wildlife. In this study, 1,019 ticks were identified on 221 hosts spanning 27 wildlife species. Next-generation sequencing was used to lify the V1-2 hypervariable region of the bacterial 16S rRNA gene from 238 ticks Amblyomma triguttatum (n=6), Bothriocroton auruginans (n=11), Bothriocroton concolor (n=20), Haemaphysalis bancrofti (n=10), Haemaphysalis bremneri (n=4), Haemaphysalis humerosa (n=13) , Haemaphysalis longicornis (n=4), Ixodes antechini (n=29), Ixodes australiensis (n=26), Ixodes fecialis (n=13), Ixodes holocyclus (n=37), Ixodes myrmecobii ( n =1), Ixodes ornithorhynchi (n=10), Ixodes tasmani (n=51) and Ixodes trichosuri (n=3). After bioinformatic analyses, over 14 million assigned bacterial sequences revealed the presence of recently described bacteria ‘ Candidatus Borrelia tachyglossi’, ‘ Candidatus Neoehrlichia australis’, ‘ Candidatus Neoehrlichia arcana’ and ‘ Candidatus Ehrlichia ornithorhynchi’. Furthermore, three novel Anaplasmataceae species were identified in the present study including a Neoehrlichia sp. in I. australiensis and I. fecialis collected from quenda ( Isoodon fusciventer ) (Western Australia), an Anaplasma sp. from one B. concolor from echidna ( Tachyglossus aculeatus ) (New South Wales), and an Ehrlichia sp. from a single I. fecialis parasitising a quenda (WA). This study highlights the ersity of bacterial genera harboured within wildlife ticks, which may prove to be of medical and/or veterinary importance in the future.
Publisher: Elsevier BV
Date: 08-2001
DOI: 10.1016/S0020-7519(01)00212-0
Abstract: This study describes the complete development (from sporozoites to sporulated oocysts) of Cryptosporidium parvum (human and cattle genotypes) in the HCT-8 cell line. Furthermore, for the first time the complete life cycle was perpetuated in vitro for up to 25 days by subculturing. The long-term maintenance of the developmental cycle of the parasite in vitro appeared to be due to the initiation of the auto-reinfection cycle of C. parvum. This auto-reinfection is characterised by the production and excystation of new invasive sporozoites from thin-walled oocysts, with subsequent maintenance of the complete life cycle in vitro. In addition, thin-walled oocysts of the cattle genotype were infective for ARC/Swiss mice but similar oocysts of the human genotype were not. This culture system will provide a model for propagation of the complete life cycle of C. parvum in vitro.
Publisher: Elsevier BV
Date: 05-2018
DOI: 10.1016/J.WATRES.2018.02.005
Abstract: As part of long-term monitoring of Cryptosporidium in water catchments serving Western Australia, New South Wales (Sydney) and Queensland, Australia, we characterised Cryptosporidium in a total of 5774 faecal s les from 17 known host species and 7 unknown bird s les, in 11 water catchment areas over a period of 30 months (July 2013 to December 2015). All s les were initially screened for Cryptosporidium spp. at the 18S rRNA locus using a quantitative PCR (qPCR). Positives s les were then typed by sequence analysis of an 825 bp fragment of the 18S gene and subtyped at the glycoprotein 60 (gp60) locus (832 bp). The overall prevalence of Cryptosporidium across the various hosts s led was 18.3% (1054/5774 95% CI, 17.3-19.3). Of these, 873 s les produced clean Sanger sequencing chromatograms, and the remaining 181 s les, which initially produced chromatograms suggesting the presence of multiple different sequences, were re-analysed by Next- Generation Sequencing (NGS) to resolve the presence of Cryptosporidium and the species composition of potential mixed infections. The overall prevalence of confirmed mixed infection was 1.7% (98/5774), and in the remaining 83 s les, NGS only detected one species of Cryptosporidium. Of the 17 Cryptosporidium species and four genotypes detected (Sanger sequencing combined with NGS), 13 are capable of infecting humans C. parvum, C. hominis, C. ubiquitum, C. cuniculus, C. meleagridis, C. canis, C. felis, C. muris, C. suis, C. scrofarum, C. bovis, C. erinacei and C. fayeri. Oocyst numbers per gram of faeces (g
Publisher: Elsevier BV
Date: 03-2012
DOI: 10.1016/J.VETPAR.2011.08.016
Abstract: On two separate s ling occasions, faecal s les were collected from lambs (2-5 months of age) grazing pasture on two separate sheep farms in southern Western Australia. Live weight, body condition score (BCS), faecal consistency score (FCS) and faecal dry matter percentage (DM%) were measured. Faecal s les were screened by PCR for Cryptosporidium (18S rRNA, actin and 60 kDa glycoprotein [gp60] loci), Giardia duodenalis (glutamate dehydrogenase [gdh] and β-giardin) and patent strongylid nematode infections (ITS-2 nuclear ribosomal DNA for Haemonchus contortus, Teladorsagia circumcincta, Trichostrongylus spp. Chabertia ovina and Oesophagostomum spp.). Faecal worm egg counts (WECs) were performed using a modified McMaster WEC technique. The WECs were adjusted for FCS and transformed using log(10)(adjusted WEC+25) prior to statistical analyses. Cryptosporidium, Giardia and Trichostrongylus spp. detected by PCR were associated with an increased risk of non-pelleted faeces (FCS ≥ 3.0) for both flocks. Cryptosporidium-positive lambs were 2.8-11.6 times more likely to have non-pelleted faeces and Giardia-positive lambs were 2.4-14.0 times more likely to have non-pelleted faeces compared to lambs negative for each respective parasite. Lambs positive for both Cryptosporidium and Giardia were 2.9-11.8 times more likely to have non-pelleted faeces than lambs positive for only one or neither of these parasites. Mixed internal parasite infections were found to have greater impacts on FCS and BCS than single infections. A higher number of internal parasites detected per lamb was associated with lower BCS and more loose faeces. The relationship between parasite detection and live weight or growth rate were inconsistent for both flocks. Adjusted WEC was correlated with FCS and faecal DM% for one flock only, although little or no correlation was found with live weight and growth rate for both flocks. Cryptosporidium ubiquitum and Cryptosporidium parvum were the most prevalent Cryptosporidium species isolated in the two flocks. Giardia assemblage E was the most commonly isolated genotype assemblage from both flocks, while assemblage A was isolated almost as frequently as assemblage E in the one flock. One flock was a potential source of zoonotic Cryptosporidium and the other flock was a potential source of zoonotic Giardia.
Publisher: Wiley
Date: 29-11-2011
DOI: 10.1111/J.1469-8137.2011.03955.X
Abstract: • Nonnative species may change ecosystem functionality at the expense of native species. Here, we examine the similarity of functional traits of native and nonnative submersed aquatic plants (SAP) in an aquatic ecosystem. • We used field and airborne imaging spectroscopy and isotope ratios of SAP species in the Sacramento-San Joaquin Delta, California (USA) to assess species identification, chlorophyll (Chl) concentration, and differences in photosynthetic efficiency. • Spectral separability between species occurs primarily in the visible and near-infrared spectral regions, which is associated with morphological and physiological differences. Nonnatives had significantly higher Chl, carotene, and anthocyanin concentrations than natives and had significantly higher photochemical reflectance index (PRI) and δ(13) C values. • Results show nonnative SAPs are functionally dissimilar to native SAPs, having wider leaf blades and greater leaf area, dense and evenly distributed vertical canopies, and higher pigment concentrations. Results suggest that nonnatives also use a facultative C(4) -like photosynthetic pathway, allowing efficient photosynthesis in high-light and low-light environments. Differences in plant functionality indicate that nonnative SAPs have a competitive advantage over native SAPs as a result of growth form and greater light-use efficiency that promotes growth under different light conditions, traits affecting system-wide species distributions and community composition.
Publisher: JSTOR
Date: 06-1999
DOI: 10.2307/3285789
Publisher: Elsevier BV
Date: 06-2016
Publisher: Cambridge University Press (CUP)
Date: 09-05-2017
DOI: 10.1017/S0031182017000439
Abstract: Little is known about the genetic ersity of the protozoan parasite, Giardia duodenalis, infecting humans in Queensland, Australia. The present study typed 88 microscopically Giardia -positive isolates using assemblage-specific primers at the triose phosphate isomerase ( tpi ) gene and sequenced a subset of isolates at the glutamate dehydrogenase ( gdh ) gene ( n = 30) and tpi locus ( n = 27). Using the tpi -assemblage specific primers, G. duodenalis assemblage A and assemblage B were detected in 50% (44/88) and 38·6% (34/88) of s les, respectively. Mixed infections with assemblages A and B were identified in 4·5% (4/88) and assemblage E was identified in 6·8% (6/88) of s les. Sequence analysis at the gdh and tpi loci also confirmed the presence of assemblage E in these isolates. Cyst numbers per gram of feces (g −1 ) were determined using quantitative polymerase chain reaction and of the isolates that were typed as assemblage E, cyst numbers ranged 13·8–68·3 × 10 6 cysts g −1 . This is the first report of assemblage E in humans in Australia, indicating that in certain settings, this assemblage may be zoonotic.
Publisher: Public Library of Science (PLoS)
Date: 28-12-2015
Publisher: Elsevier BV
Date: 03-2012
DOI: 10.1016/J.EXPPARA.2011.12.009
Abstract: The dispute on the validity of Cryptosporidium pestis and Cryptosporidium tyzzeri origins from the uncertainty on the identity of Cryptosporidium parvum described by Tyzzer in 1912 and the interpretation of the Principal of Priority of the International Code of Zoological Nomenclature (ICZN). Using a rigid interpretation of the Principal of Priority, one researcher proposed to rename C. parvum as C. pestis and retain C. parvum for Cryptosporidium mouse genotype I on the basis that Tyzzer was probably describing mouse genotype I. However, the ICZN clearly states that the Principle of Priority is to be used to promote stability and is not intended to upset a long-accepted name. Because mice are known to be naturally infected with C. parvum, and the 1985 taxonomic re-description of C. parvum for bovine and human isolates is accepted by almost all Cryptosporidium researchers, the prevailing name C. parvum for the species infective to calves and humans must be retained to avoid confusion. The designation of C. tyzzeri for the mouse genotype I brings further clarity to the taxonomy of Cryptosporidium spp. in humans, cattle, and domestic mice.
Publisher: Elsevier BV
Date: 07-2017
Publisher: Elsevier BV
Date: 07-2017
Abstract: Enteric parasites are major contributors to the global diarrhoeal disease load, infecting >67.2 million people. Their prevalence and clinical impact, however, are underestimated due to lack of adequate detection, which is largely still based on microscopy, particularly in developing countries. New commercially available enteric panel assays, which detect parasites (as well as bacteria and/or viruses) using multiplex PCR, offer enhanced sensitivity and specificity as well as the ability to detect mixed infections, and will play an important role in epidemiological surveillance and outbreak investigations. A major limitation of these technologies, however, particularly for developing countries, is the costs involved. Emerging technologies for low-resource, point-of-care (POC) settings have the potential to dramatically improve the cost and accuracy of enteric parasite detection in the future.
Publisher: Elsevier BV
Date: 09-2008
DOI: 10.1016/J.EXPPARA.2008.04.023
Abstract: The complete development of Cryptosporidium in host cell-free medium first described in 2004, represented a significant advance that can facilitate many aspects of Cryptosporidium research. A current limitation of host cell-free cultivation is the difficulty involved in visualising the life-cycle stages as they are very small in size, morphologically difficult to identify and dispersed throughout the media. This is in contrast to conventional cell culture methods for Cryptosporidium, where it is possible to focus on the host cells and view the foci of infection on the host cells. In the present study, we compared three specific and three non-specific techniques for visualising Cryptosporidium parvum life-cycle stages in cell-free culture antibody staining using anti-sporozoite and anti-oocyst wall antibodies (Sporo-Glo and Crypto Cel), fluorescent in-situ hybridization (FISH) using a Cryptosporidium specific rRNA oligonucleotide probe and the non-specific dyes Texas Red, carboxyfluorescein diacetate succinimidyl ester (CFSE) and 4,6' diamino-2-phenylindole dihydrochloride (DAPI). Results revealed that a combination of Sporo-Glo and Crypto Cel staining resulted in easy and reliable identification of all life-cycle stages.
Publisher: Elsevier BV
Date: 04-2017
DOI: 10.1016/J.VETPAR.2017.01.027
Abstract: The genus term Isospora is now applied specifically to parasites of birds, with the term Cystoisospora preferred for parasites which infect mammals. Isospora is a common parasitic coccidian in birds worldwide, especially in passerine birds, in which it can cause systemic coccidiosis. The complete mitochondrial genome sequences from two recently identified Isospora species Isospora serinuse in a domestic canary and Isospora manorinae in a yellow-throated miner, were sequenced and compared with those of other closely related coccidian species. The complete mitochondrial genome sequence for Isospora serinuse is 6260bp in size and 6223bp for Isospora manorinae. The mitochondrial genomes of Isospora serinuse and Isospora manorinae include three protein-coding genes (COI, COIII and CytB), 19 LSU and 14 SSU rDNA fragments, including one newly identified putative LSU fragment in Isospora sp. The arrangement of coding regions in these two Isospora species were identical to that of available Isospora sp. and Eimeria spp. mitochondrial genomes and the start codon usage for protein coding genes was conservative. Phylogenetic analysis of the mt genome of the two Isospora species based on the three coding regions further support that the monophyletic nature of avian Isospora.
Publisher: Wiley
Date: 2000
DOI: 10.1111/J.1550-7408.2000.TB00016.X
Abstract: A new species of Cryptosporidium is described from the feces of domestic cattle, Bos taurus. Oocysts are structurally similar to those of Cryptosporidium muris described from mice but are larger than those of Cryptosporidium parvum. Oocysts of the new species are ellipsoidal, lack sporocysts, and measure 7.4 x 5.5 microm (range, 6.0-8.1 by 5.0-6.5 microm). The length to width ratio is 1.35 (range, 1.07-1.50). The colorless oocyst wall is < 1 microm thick, lacks a micropyle, and possesses a longitudinal suture at one pole. A polar granule is absent, whereas an oocyst residuum is present. Oocysts were passed fully sporulated and are not infectious to outbred, inbred immunocompetent or immunodeficient mice, chickens or goats. Recent molecular analyses of the rDNA 18S and ITS1 regions and heat-shock protein 70 (HSP-70) genes demonstrate this species to be distinct from C. muris infecting rodents. Based on transmission studies and molecular data, we consider the large form of Cryptosporidium infecting the abomasum of cattle to be a new species and have proposed the name Cryptosporidium andersoni n. sp. for this parasite.
Publisher: Elsevier BV
Date: 11-2008
Publisher: Elsevier BV
Date: 08-2016
DOI: 10.1016/J.VETPAR.2016.06.027
Abstract: Cryptosporidium is a protozoan parasite that infects a wide range of hosts, yet relatively little is known about the epidemiology of cryptosporidiosis in fish. Here we report a disseminated Cryptosporidium infection in a male Koi carp (Cyprinus carpio), with parasite stages identified deep within the epithelium of the intestine, kidneys, spleen, liver and gills causing severe granulomatous inflammatory lesions. Molecular characterization at two loci 18S ribosomal RNA (rRNA) and actin, revealed this to be a novel Cryptosporidium genotype, most closely related to Cryptosporidium molnari.
Publisher: Elsevier BV
Date: 04-2016
DOI: 10.1016/J.EXPPARA.2016.01.013
Abstract: A new Isospora (Apicomplexa:Eimeriidae) species is described from a single yellow-throated miner bird (Manorina flavigula) (subspecies M. f. wayensis) in Western Australia. Sporulated oocysts (n = 32) of this isolate are spherical to subspherical, 22.8 (20.3-23.8) × 18.3 (17.7-18.7) μm, with a shape index (length/width) of 1.25 (1.2-1.3) and a smooth and bilayered oocyst wall, 1.3 μm thick (outer layer 0.9 μm, inner 0.4 μm). A polar granule is present, but the micropyle and oocyst residuum are absent. The sporocysts are lemon-shaped, 15.5 (14.6-15.8) × 9.5 (9.5-10.2) μm, with a shape index of 1.6. Stieda and substieda bodies are present, the Stieda body being knob-like and the substieda body being subspherical-shaped. A sporocyst residuum is present and composed of numerous granules of different size scattered among the sporozoites, a spheroid or subspheroid refractile body is present in the sporozoite. Morphologically, the oocysts from this isolate are different from those of all known valid Isospora spp. Molecular analysis was conducted at 3 loci the 18S and 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) gene. At the 18S locus, this new isolate exhibited 99.2% similarity to Isospora gryphoni and three other Isospora spp. Further analysis of a subgroup of 300 bp long 18S sequences (8), including Isospora anthochaerae was conducted. This new isolate grouped in a clade with I. anthochaerae and exhibited 99.3% similarity. At the 28S locus, this new isolate grouped with I. anthochaerae with which it shared 99.1% similarity. At the COI locus, this new isolate exhibited 96.8% similarity to Isospora sp. JCI-2015 from a spectacled warbler (Sylvia conspicillata) in Spain. Further analysis from a subgroup of shorter COI sequences (n = 13) was performed and this new isolate exhibited 99.1% similarity to I. anthochaerae. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora manorinae n. sp. after its host, the yellow-throated miner (Manorina flavigula wayensis).
Publisher: Elsevier BV
Date: 03-2008
Publisher: Cambridge University Press (CUP)
Date: 06-05-2009
DOI: 10.1017/S0031182009005927
Abstract: Little is known of the prevalence and life-cycle of trypanosomes in mammals native to Australia. Native Australian trypanosomes have previously been identified in marsupials in the eastern states of Australia, with one recent report in brush-tailed bettongs ( Bettongia penicillata ), or woylie in Western Australia in 2008. This study reports a novel Trypanosoma sp. identified in blood smears, from 7 critically endangered Gilbert's potoroos ( Potorous gilbertii ) and 3 quokkas ( Setonix brachyurus ) in Western Australia. Trypanosomes were successfully cultured in vitro and showed morphological characteristics similar to members of the subgenus Herpetosoma . Phylogenetic analysis of 18S rRNA gene sequences identified 2 different novel genotypes A and B that are closely related to trypanosomes previously isolated from a common wombat ( Vombatus ursinus ) in Victoria, Australia. The new species is proposed to be named Trypanosoma copemani n. sp.
Publisher: American Society of Parasitologists
Date: 12-2001
Publisher: Elsevier BV
Date: 11-2020
Publisher: American Society of Parasitologists
Date: 04-2000
Publisher: American Society for Microbiology
Date: 12-2000
DOI: 10.1128/AEM.66.12.5499-5502.2000
Abstract: Nucleotide sequences of the Cryptosporidium oocyst wall protein (COWP) gene were obtained from various Cryptosporidium spp. ( C. wrairi , C. felis , C. meleagridis , C. baileyi , C. andersoni , C. muris , and C. serpentis ) and C. parvum genotypes (human, bovine, monkey, marsupial, ferret, mouse, pig, and dog). Significant ersity was observed among species and genotypes in the primer and target regions of a popular diagnostic PCR. These results provide useful information for COWP-based molecular differentiation of Cryptosporidium spp. and genotypes.
Publisher: Springer Science and Business Media LLC
Date: 25-06-2015
Publisher: Wiley
Date: 02-2001
DOI: 10.1111/J.1751-0813.2001.TB10708.X
Abstract: Cryptosporidium parvum, an intestinal coccidian parasite, was isolated from faeces and intestinal biopsies of a 9-week-old puppy with acute parvoviral gastroenteritis. Gene sequence analysis identified a Cryptosporidium genotype not previously recorded in Australia. The puppy recovered after treatment with crystalloid fluids, synthetic and natural colloids and jejunostomy tube feeding.
Publisher: Cambridge University Press (CUP)
Date: 11-1999
DOI: 10.1017/S0031182099004102
Abstract: The development of molecular diagnostic methods, particularly those utilizing PCR for the detection of parasitic protozoa will contribute greatly to the identification and control of these pathogens, by increasing speed of diagnosis, specificity and sensitivity, reproducibility and ease of interpretation. PCR methods are not without their problems however, and there is a need for laboratory procedures to be refined before PCR-based assays are accepted as the tools of choice for the routine detection of protozoan parasites. The application of PCR detection to various parasites is discussed.
Publisher: Elsevier BV
Date: 12-2011
DOI: 10.1016/J.VETPAR.2011.05.050
Abstract: On two extensive sheep farms in southern Western Australia, 111 (Farm A) and 124 (Farm B) female crossbred lambs (2-6 weeks old) were randomly selected and in idually identified using ear tags (a numbered tag and radio-frequency tag) at marking. On five separate occasions, faecal s les were collected and live weight, body condition score (BCS), faecal consistency score (FCS), breech fleece faecal soiling score and faecal dry matter percentage (DM%) were recorded. Lamb hot carcase weight (HCW) and dressing percentage were measured at slaughter. Faecal s les were screened by PCR for Cryptosporidium (18S rRNA, actin and 60 kDa glycoprotein [gp60] loci), Giardia duodenalis (glutamate dehydrogenase [gdh] and triosephosphate isomerise [tpi]) and C ylobacter jejuni (16S rRNA). Observation of Eimeria oocysts and faecal worm egg counts (WECs) were performed using a modified McMaster technique. The WECs were adjusted for FCS for analyses. Faecal s les were screened for patent strongylid infections using PCR (specifically ITS-2 nuclear ribosomal DNA for Teladorsagia circumcincta, Trichostrongylus spp. and Haemonchus contortus). Lambs positive for Cryptosporidium at least once had lighter HCWs by 1.25 kg (6.6%) (P=0.029) and 1.46 kg (9.7%) (P<0.001) compared to lambs never positive for Cryptosporidium for Farms A and B respectively. Similarly, dressing percentages were 1.7% (P=0.022) and 1.9% (P<0.001) lower in Cryptosporidium-positive lambs on Farms A and B respectively. Lambs positive for Giardia at least once had 0.69 kg (P<0.001) lighter HCWs and 1.7% (P<0.001) lower dressing percentages compared to lambs never positive for Giardia on Farm B only. Cryptosporidium-positive lambs at the second s ling were 4.72 (P=0.010) and 3.84 (P=0.002) times more likely to have non-pelleted faeces compared to Cryptosporidium-negative lambs for Farms A and B respectively. Breech fleece faecal soiling scores of Cryptosporidium-positive lambs were 3.36 (P=0.026) and 2.96 (P=0.047) times more likely to be moderate to severe (scores 3-5), compared to negative lambs at the second s ling for Farms A and B respectively. Live weight, growth rate and BCS were inconsistently associated with protozoa detection across different s lings and farms. Adjusted WEC was correlated positively with FCS and negatively with faecal DM%, differing between s ling occasions and farms. C ylobacter jejuni prevalence was very low (<1%). Adjusted WEC were not correlated with carcase attributes, growth rates or live weights. This study is the first to quantify productivity consequences of naturally acquired protozoa infections in lambs managed under extensive farming conditions.
Publisher: Elsevier BV
Date: 2018
DOI: 10.1016/J.IJPARA.2017.09.004
Abstract: Foodborne illness, the majority of which is caused by enteric infectious agents, costs global economies billions of dollars each year. The protozoan parasite Cryptosporidium is particularly suited to foodborne transmission and is responsible for >8 million cases of foodborne illness annually. Procedures have been developed for sensitive detection of Cryptosporidium oocysts on fresh produce and molecular diagnostic assays have been widely used in case linkages and infection source tracking, especially during outbreak investigations. The integrated use of advanced diagnostic techniques with conventional epidemiological studies is essential to improve our understanding of the occurrence, source and epidemiology of foodborne cryptosporidiosis. The implementation of food safety management tools such as Good Hygienic Practices (GHP), Hazard Analysis and Critical Control Points (HACCP), and Quantitative Microbial Risk Assessment (QMRA) in industrialised nations and Water, Sanitation, and Hygiene (WASH) in developing countries is central for prevention and control and foodborne cryptosporidiosis in the future.
Publisher: Elsevier BV
Date: 03-2006
Abstract: The Australian Research Council (ARC) and the National Health and Medical Research Council (NHMRC) Research Network for Parasitology will focus and coordinate the fundamental, strategic and applied parasitology research in Australia. It will raise the standing of Australia in the field, assist in the community understanding of parasitology, and maintain and improve the capacity of Australia to keep its stock, crops, wildlife and people free from disease. On an international scale, the ARC/NHMRC Network will work with other countries to develop new technologies for the detection and control of parasites.
Publisher: Cambridge University Press (CUP)
Date: 21-07-2010
DOI: 10.1017/S0031182010000971
Abstract: Trypanosoma irwini was previously described from koalas and we now report the finding of a second novel species, T. gilletti , as well as the extension of the host range of Trypanosoma copemani to include koalas. Phylogenetic analysis at the 18S rDNA and gGAPDH loci demonstrated that T. gilletti was genetically distinct with a genetic distance (± s.e .) at the 18S rDNA locus of 2·7±0·5% from T. copemani (wombat). At the gGAPDH locus, the genetic distance (± s.e. ) of T. gilletti was 8·7±1·1% from T. copemani (wombat). Trypanosoma gilletti was detected using a nested trypanosome 18S rDNA PCR in 3/139 (∼2%) blood s les and in 2/29 (∼7%) spleen tissue s les from koalas whilst T. irwini was detected in 72/139 (∼52%) blood s les and T. copemani in 4/139 (∼3%) blood s les from koalas. In addition, naturally occurring mixed infections were noted in 2/139 (∼1·5%) of the koalas tested.
Publisher: Elsevier BV
Date: 07-2015
DOI: 10.1016/J.EXPPARA.2015.04.019
Abstract: A new species, Eimeria collieie n. sp., is described from the western long-necked turtle (Chelodina colliei). Sporulated oocysts (n = 35) are spherical to subspherical, with colourless single layer oocyst wall, 0.6 ± 0.2 (0.4-0.7) µm thick. Oocyst with elongated ellipsoid sporocysts. Oocyst length, 29.8 ± 0.4 (28.2-31.0) µm oocyst width, 29.4 ± 0.3 (28.0-30.8) µm oocyst length/width (L/W) ratio, 1.0 ± 0.03 (1.0-1.05). Micropyle, oocyst residuum and polar granule were absent. Sporocysts with sporocyst residuum and 2 sporozoites. Sporocyst length, 21.6 ± 0.4 (21.2-22.0) µm sporocyst width, 6.0 ± 0.3 (5.7-6.3) µm sporocyst L/W ratio, 3.6 ± 0.2 (3.4-3.8). Stieda, parastieda and substieda bodies were absent. Sporozoite length, 14.0 ± 0.2 (13.8-14.2) µm sporozoite width, 2.6 ± 0.2 (2.4-2.8) µm sporozoite L/W ratio, 5.46 ± 0.10 (5.4-5.6). Molecular analysis was conducted at three loci: the 18S and 28S ribosomal RNA (rRNA), and the mitochondrial cytochrome oxidase gene (COI). At the 18S rRNA locus, E. collieie n. sp. shared 96.4% and 98.3% genetic similarity to E. ranae (GenBank accession number: EU717219) and E. arnyi (AY613853) respectively. At the 28S rRNA locus, E. collieie n. sp. shared 91.6% genetic similarity to E. papillata (GenBank accession number: GU593706) and phylogenetic analysis at this locus placed E. collieie n. sp. in aseparateclade. At the COI locus, E. collieie n. sp. shared 92.7% genetic similarity to Eimeria setonicis (GenBankaccession number: KF225638) from a quokka (Setonix brachyurus) in Western Australia. Reptile-derived sequences were not available for the 28S rRNA and the COI loci. Based on morphological and molecular data, this isolate is a new species of coccidian parasite that, to date, has only been found in western long-necked turtles.
Publisher: Wiley
Date: 03-2000
DOI: 10.1111/J.1751-0813.2000.TB10589.X
Abstract: To perform a morphological and genetic characterisation of a Cryptosporidium infection in an Indian ring-necked parrot (Psittacula krameri) and to compare this with C meleagridis from a turkey. Tissue and intestinal sections from an Indian ring-necked parrot were examined microscopically for Cryptosporidium. The organism was also purified from the crop and intestine, the DNA extracted and a portion of the 18S rDNA gene lified, sequenced and compared with sequence and biological information obtained for C meleagridis from a turkey as well as sequence information for other species of Cryptosporidium. Morphological examination of tissue sections from an Indian ring-necked parrot revealed large numbers of Cryptosporidium oocysts attached to the apical border of enterocytes lining the intestinal tract. Purified Cryptosporidium oocysts measured about 5.1 x 4.5 microns, which conformed morphologically to C meleagridis. The sequence obtained from this isolate was identical to sequence information obtained from a C meleagridis isolate from a turkey. Cryptosporidium meleagridis was detected in an Indian ring-necked parrot using morphological and molecular methods. This is the first time that this species of Cryptosporidium has been reported in a non-galliform host and extends the known host range of C meleagridis.
Publisher: Elsevier BV
Date: 2021
Publisher: Elsevier BV
Date: 02-2007
Publisher: Informa UK Limited
Date: 17-04-2023
Publisher: Elsevier BV
Date: 03-2001
DOI: 10.1016/S0020-7519(00)00164-8
Abstract: Avian isolates of Cryptosporidium species from different geographic locations were sequenced at two loci, the 18S rRNA gene and the heat shock gene (HSP-70). Phylogenetic analysis of the sequence data provided support for the existence of a new avian species of Cryptosporidium infecting finches and a second species infecting a black duck. The identity of Cryptosporidium baileyi and Cryptosporidium meleagridis as valid species was confirmed. Also, C. baileyi was identified in a number of isolates from the brown quail extending the host range of this species.
Publisher: American Association for the Advancement of Science (AAAS)
Date: 31-05-2019
Abstract: Improving costs and scale reflect looming opportunities
Publisher: Elsevier BV
Date: 06-2016
DOI: 10.1016/J.PARINT.2015.12.005
Abstract: The quokka, Setonix brachyurus, is a vulnerable, small marsupial endemic to Western Australia. Blood s les were collected from quokkas from three different geographical locations Two Peoples Bay, Bald Island and Rottnest Island. The overall prevalence of trypanosomes by nested PCR at the 18S ribosomal RNA gene was 57.3% (63/110) with prevalences of 91.4%, 85.3% and 4.9% respectively for Two Peoples Bay, Bald Island and Rottnest Island. Phylogenetic analysis conducted on 47 18S PCR positives identified two Trypanosoma copemani genotypes, with T. copemani genotype B, the most prevalent genotype infecting quokka populations from the three locations with an overall prevalence of 51.8% (24/47) compared to 34% for T. copemani genotype A (16/47). The overall prevalence of mixed T. copemani genotype A and B infections was 14.9% (7/47). Phylogenetic analysis of 26 quokka isolates at the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) locus, largely supported the 18S analysis but identified a mixed infection in one quokka isolate (Q4112-4117 from Two Peoples Bay). T. copemani genotype B has previously only been isolated from quokkas and the Gilbert's potoroo whereas T. copemani genotype A has a wide host range and may be pathogenic. Further work is required to determine the clinical impact of T. copemani on marsupial populations.
Publisher: Elsevier BV
Date: 03-2007
DOI: 10.1016/J.VETPAR.2006.09.022
Abstract: Canine piroplasmosis is an emerging disease worldwide, with multiple species of piroplasm now recognised to infect dogs. A nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was developed for the detection and differentiation of each of the piroplasm species currently known to infect dogs on the basis of the 18S ribosomal RNA gene. The assay can potentially lify and discriminate between Theileria annae, Theileria equi, Babesia conradae, Babesia gibsoni, Babesia sp. (Coco) and each of the Babesia canis subspecies. Non-canine piroplasm species can also potentially be detected using the described assay, however lification of Neospora caninum was also observed. The PCR was found to have a high detection limit, capable of detecting a 2.7x10(-7)% parasitaemia or the equivalent of 1.2 molecules of target DNA when using DNA extracted from whole EDTA blood and detected a parasitaemia of 2.7x10(-5)% using blood applied to both Flinders Technology Associates (FTA) cards and IsoCodetrade mark Stix. The application of blood s les to filter paper may greatly assist in piroplasm identification in regions of the world where local technologies for molecular characterisation are limited. The assay reported here has the potential to be standardised for routine screening of dogs for piroplasmosis.
Publisher: Elsevier BV
Date: 07-2000
DOI: 10.1016/S0169-4758(00)01699-9
Abstract: There is controversy in the taxonomy of Cryptosporidium parasites and the public health significance of Cryptosporidium isolates from various animals. Recent advances in molecular characterization of Cryptosporidium parasites have allowed the re-examination of species structure of the genus Cryptosporidium. Non-parvum Cryptosporidium spp and new C. parvum genotypes in immunocompromised humans can now be clearly detected. In this article, Lihua Xiao and colleagues summarize the current biological and molecular evidence for different Cryptosporidium spp, and the public health importance of these species and new C. parvum genotypes.
Publisher: American Society of Parasitologists
Date: 04-2008
DOI: 10.1645/GE-1344.1
Publisher: Elsevier BV
Date: 06-2015
DOI: 10.1016/J.EXPPARA.2015.03.022
Abstract: Trypanosoma copemani is known to be infective to a variety of Australian marsupials. Characterisation of this parasite revealed the presence of stercorarian-like life-cycle stages in culture, which are similar to T. rangeli and T. cruzi. The blood incubation infectivity test (BIIT) was adapted and used to determine if T. copemani, like T. cruzi and T. rangeli, has the potential to grow in the presence of human serum. To eliminate any effects of anticoagulants on the complement system and on human high density lipoprotein (HDL), only fresh whole human blood was used. Trypanosoma copemani was observed by microscopy in all human blood cultures from day 5 to day 19 post inoculation (PI). The mechanism for normal human serum (NHS) resistance in T. copemani is not known. The results of this study show that at least one native Australian trypanosome species may have the potential to be infective for humans.
Publisher: Cambridge University Press (CUP)
Date: 26-06-2015
DOI: 10.1017/S095026881400106X
Abstract: Cryptosporidiosis is a gastroenteric disease caused by the protozoan parasite Cryptosporidium , which manifests primarily as watery diarrhoea. Transmitted via the faecal–oral route, infection with the parasite can occur through ingestion of water, food or other fomites contaminated with its infective oocyst stage. In the months of November and December 2012, there were 18 notified cases of cryptosporidiosis from Broome, Western Australia. The 5-year average for the Kimberley region for this period is case. Interviews conducted by Broome local government staff on the notified cases revealed that 11/18 cases had been swimming at the Broome public swimming pool. Molecular analyses of extracted DNA performed on 8/18 microscopy-positive faecal s les from interviewed cases and three water s les from different locations at the hypervariable glycoprotein 60 ( gp60 ) gene, identified the C. hominis IbA10G2 subtype in all human s les and one water s le.
Publisher: American Society for Microbiology
Date: 07-2003
DOI: 10.1128/AEM.69.7.3970-3974.2003
Abstract: Over a 3-year period, a total of 646 fecal s les from pigs in 22 indoor and outdoor herds from Western Australia were screened for Cryptosporidium spp. by microscopy. Results revealed that 39 of 646 s les (6.03%) were positive for Cryptosporidium. Cryptosporidium was much more common in outdoor herds (17.2%) than in indoor herds (0.5%) and was more common in animals between the ages of 5 and 8 weeks (69.2%) than in younger animals ( P 0.0001). Molecular characterization of the positive s les at the 18S ribosomal DNA locus identified two distinct genotypes of Cryptosporidium : the previously identified pig genotype I and a novel pig genotype (pig genotype II), both of which warrant species status.
Publisher: Springer Science and Business Media LLC
Date: 20-03-2019
Publisher: Elsevier BV
Date: 04-2022
Abstract: Zoonotic cryptosporidiosis is a major public health problem in industrialized nations in those countries it is caused mainly by Cryptosporidium parvum IIa subtypes that are prevalent in dairy calves. Because of the short history of intensive animal farming in China, strains of C. parvum are found only on some dairy farms in this country and are the IId subtypes. However, the prevalence of C. parvum is increasing rapidly, with IIa subtypes recently detected in a few grazing animals, and both IIa and IId subtypes are emerging in humans. As animal farming intensifies, China may follow in the footsteps of industrialized nations where zoonotic cryptosporidiosis is r ant. One Health and biosecurity measures are urgently needed to slow down the dispersal of autochthonous IId subtypes and imported IIa subtypes.
Publisher: Elsevier BV
Date: 08-2021
DOI: 10.1016/J.MEEGID.2021.104859
Abstract: Cryptosporidium is an important protozoan parasite and due to its resistance to chlorine is a major cause of swimming pool-associated gastroenteritis outbreaks. The present study combined contact tracing and molecular techniques to analyse cryptosporidiosis cases and outbreaks in Western Australia in 2019 and 2020. In the 2019 outbreak, subtyping at the 60 kDa glycoprotein (gp60) gene identified 89.0% (16/18) of s les were caused by the C. hominis IdA15G1 subtype. Amplicon next generation sequencing (NGS) at the gp60 locus identified five C. hominis IdA15G1 subtype s les that also had C. hominis IdA14 subtype DNA, while multi locus sequence typing (MLST) analysis on a subset (n = 14) of C. hominis s les identified three IdA15G1 s les with a 6 bp insertion at the end of the trinucleotide repeat region of the cp47 gene. In 2020, 88.0% (73/83) of s les typed were caused by the relatively rare C. hominis subtype IbA12G3. Four mixed infections were observed by NGS with three IdA15G1/ IdA14 mixtures and one C. parvum IIaA18G3R1 s le mixed with IIaA16G3R1. No genetic ersity using MLST was detected. Epidemiological and molecular data indicates that the outbreaks in 2019 and 2020 were each potentially from swimming pool point sources and a new C. hominis subtype IbA12G3 is emerging in Australia. The findings of the present study are important for understanding the introduction and transmission of rare Cryptosporidium subtypes to vulnerable populations.
Publisher: Springer Science and Business Media LLC
Date: 06-01-2021
Publisher: Elsevier BV
Date: 08-2017
DOI: 10.1016/J.VPRSR.2017.05.001
Abstract: Faecal shedding of Eimeria by captured rangeland goats (Capra hircus) was investigated using a longitudinal observational study. Faecal s les were collected from 125 male goats on four occasions. The first s ling occurred following capture and transport, immediately after arrival at a commercial goat depot (feedlot) in Western Australia, with subsequent 3 s le collections occurring at one month intervals thereafter. Goats were composite breed and aged approximately 9-12months on arrival at the feedlot. Prevalence and shedding intensity (faecal oocyst concentration) for Eimeria were determined using qPCR. Species were identified from in idual oocysts (isolated using micromanipulation) using molecular analysis at two loci, specifically 18S rRNA and mitochondrial cytochrome oxidase gene (COI), and confirmed by microscopy. Longitudinal prevalence (animals positive at least once) for Eimeria spp. by qPCR was 90.4%, with 60% goats shedding Eimeria spp. on more than one occasion. Point prevalence (prevalence at a single s ling occasion) ranged from 2.4% (fourth s ling) to 70.4% (second s ling). Three species were identified at the 18S rRNA locus and confirmed by microscopy: E. christenseni (longitudinal prevalence for single infection 34.4%), E. hirci (17.6%) and E. arloingi (8.8%) over the four s le collections. Mixed infections were identified in 56.8% goats (longitudinal prevalence). 18S rRNA sequences from E. christenseni and E. hirci were 100% homologous with ovine E. ahsata and E. crandallis respectively, and E. arloingi was 100% similar to caprine E. arloingi. At the COI locus, E. christenseni, E. hirci and E. arloingi grouped separately, and were closely related to ovine E. ahsata, with genetic similarities of 96.5%, 92.6% and 91.4% respectively. This is the first report for molecular characteristics of caprine-derived Eimeria spp. using a combination of 18S rRNA and COI. Molecular techniques can be used to identify Eimeria spp. in goat faecal s les, specifically through characterization at 18S locus and other gene loci when used in parallel. Molecular techniques offer some advantages over microscopy for identification of Eimeria species, particularly with respect to precision.
Publisher: Cambridge University Press (CUP)
Date: 05-07-2018
DOI: 10.1017/S0950268818001607
Abstract: Cryptosporidium is a protozoan parasite that causes the diarrhoeal disease, cryptosporidiosis. Although many species have been identified, the majority of human disease worldwide is caused by two species Cryptosporidium parvum and Cryptosporidium hominis. In Australia, data from the National Notifiable Diseases Surveillance System (NNDSS) show that cryptosporidiosis outbreaks occur every few years. To better understand the transmission, trends and nature of cryptosporidiosis outbreaks in Western Australia, epidemiological and genomic data from three cryptosporidiosis outbreaks in 2003, 2007 and 2011 were reviewed. The 2007 outbreak was the largest ( n = 607) compared with the outbreaks in 2003 ( n = 404) and 2011 ( n = 355). All three outbreaks appeared to have occurred predominantly in the urban metropolitan area (Perth), which reported the highest number of case notifications increases in case notifications were also observed in rural and remote areas. Children aged 0–4 years and non-Aboriginal people comprised the majority of notifications in all outbreaks. However, in the 2003 and 2007 outbreaks, a higher proportion of cases from Aboriginal people was observed in the remote areas. Molecular data were only available for the 2007 ( n = 126) and 2011 ( n = 42) outbreaks, with C. hominis the main species identified in both outbreaks. Subtyping at the glycoprotein 60 ( gp60 ) locus identified subtype IbA10G2 in 46.3% and 89.5% of C. hominis isolates typed, respectively, in the 2007 and 2011 outbreaks, with the IdA15G1 subtype was identified in 33.3% of C. hominis isolates typed in the 2007 outbreak. The clustering of cases with the IdA15G1 subtype in the remote areas suggests the occurrence of a concurrent outbreak in remote areas during the 2007 outbreak, which primarily affected Aboriginal people. Both the C. hominis IbA10G2 and IdA15G1 subtypes have been implicated in cryptosporidiosis outbreaks worldwide its occurrence indicates that the mode of transmission in both the 2007 and 2011 outbreaks was anthroponotic. To better understand the epidemiology, sources and transmission of cryptosporidiosis in Australia, genotyping data should routinely be incorporated into national surveillance programmes.
Publisher: Springer Science and Business Media LLC
Date: 13-03-2021
Publisher: Elsevier BV
Date: 05-2019
Publisher: Elsevier BV
Date: 09-2019
DOI: 10.1016/J.MEEGID.2019.05.018
Abstract: Cryptosporidium species are a major cause of diarrhoea worldwide. In the present study, a retrospective analysis of 109 microscopically Cryptosporidium-positive faecal specimens from Western Australian patients, collected between 2015 and 2018 was conducted. Sequence analysis of the 18S rRNA and the 60 kDa glycoprotein (gp60) gene loci identified four Cryptosporidium species: C. hominis (86.2%, 94/109), C. parvum (11.0%, 12/109), C. meleagridis (1.8%, 2/109) and C. viatorum (0.9%, 1/109). Subtyping at the gp60 locus identified a total of 11 subtypes including the emergence of the previously rare C. hominis IfA12G1R5 subtype in 2017 as the dominant subtype (46.7%, 21/45). This subtype has also recently emerged as the dominant subtype in the United States but the reasons for its emergence are unknown. This is also the first report of C. viatorum in humans in Australia and a novel subtype (XVaA3g) was identified in the one positive patient.
Publisher: Elsevier BV
Date: 10-2017
DOI: 10.1016/J.VETPAR.2017.08.014
Abstract: The morphological, biological, and molecular characterisation of a new Cryptosporidium species from the guinea pig (Cavia porcellus) are described, and the species name Cryptosporidium homai n. sp. is proposed. Histological analysis conducted on a post-mortem s le from a guinea pig euthanised due to respiratory distress, identified developmental stages of C. homai n. sp. (trophozoites and meronts) along the intestinal epithelium. Molecular analysis at 18S rRNA (18S), actin and hsp70 loci was then conducted on faeces from an additional 7 guinea pigs positive for C. homai n. sp. At the 18S, actin and hsp70 loci, C. homai n. sp. exhibited genetic distances ranging from 3.1% to 14.3%, 14.4% to 24.5%, and 6.6% to 20.9% from other Cryptosporidium spp., respectively. At the 18S locus, C. homai n. sp. shared 99.1% similarity with a previously described Cryptosporidium genotype in guinea pigs from Brazil and it is likely that they are the same species, however this cannot be confirmed as actin and hsp70 sequences from the Brazilian guinea pig genotype are not available. Phylogenetic analysis of concatenated 18S, actin and hsp70 sequences showed that C. homai n. sp. exhibited 9.1% to 17.3% genetic distance from all other Cryptosporidium spp. This clearly supports the validity of C. homai n. sp. as a separate species.
Publisher: Elsevier BV
Date: 09-2017
Publisher: MDPI AG
Date: 16-12-2020
DOI: 10.3390/MICROORGANISMS8122014
Abstract: Current knowledge of Cryptosporidium species/genotypes in marine fish is limited. Following phylogenetic analysis at the 18S rDNA locus, a recent study identified six new genotypes of Cryptosporidium colonizing edible fish found in European seas. Of these, five grouped in a clade together (#Cryptofish 1–5) and one grouped separately (#Cryptofish 7). In the present study, after phylogenetic analyses of #Cryptofish1, #Cryptofish2, #Cryptofish4, #Cryptofish5 and #Cryptofish7 at the actin locus, the presence of two major clades was confirmed. In addition, when possible, longer 18S licons were generated. In conclusion, the small genetic distances between these genotypes designated as a novel marine genotype I (#Cryptofish 1-5) suggest that they may be genetic variants of the same species, while the designated novel marine genotype 2 (#Cryptofish 7) is clearly representative of a separate species.
Publisher: Elsevier BV
Date: 09-2012
DOI: 10.1016/J.PARINT.2012.02.009
Abstract: The nuclear ribosomal DNA (rDNA) region spanning 5.8S rDNA and the second internal transcribed spacer (ITS-2) of Baylisascaris schroederi isolated from the Qinling subspecies of giant panda in Shaanxi Province, China were lified and sequenced. Sequence variations in the two rDNA regions within B. schroederi and among species in the family Ascarididae were examined. The lengths of B. schroederi 5.8S and ITS-2 rDNA sequences were 156 bp and 327 bp, respectively, and no nucleotide variation was found in these two rDNA regions among the 20 B. schroederi s les examined, and these ITS-2 sequences were identical to that of B. schroederi isolated from giant panda in Sichuan province, China. The inter-species differences in 5.8S and ITS-2 rDNA sequences among members of the family Ascarididae were 0-1.3% and 0-17.7%, respectively. Phylogenetic relationships among species in the Ascarididae were re-constructed by Bayesian inference (Bayes), maximum parsimony (MP), and maximum likelihood (ML) analyses, based on combined sequences of 5.8S and ITS-2 rDNA. All B. schroederi s les clustered together and sistered to B. transfuga with high posterior probabilities/bootstrap values, which further confirmed that nematodes isolated from the Qinling subspecies of giant panda in Shaanxi Province, China represent B. schroederi. Because of the large number of ambiguously aligned sequence positions (difficulty of inferring homology by positions), ITS-2 sequence alone is likely unsuitable for phylogenetic analyses at the family level, but the combined 5.8S and ITS-2 rDNA sequences provide alternative genetic markers for the identification of B. schroederi and for phylogenetic analysis of parasites in the family Ascarididae.
Publisher: Elsevier BV
Date: 03-2010
DOI: 10.1016/J.VETPAR.2009.11.015
Abstract: Little is known about the prevalence and genotypes of Cryptosporidium in fish. The present study investigated the prevalence of Cryptosporidium species in cultured fingerlings (n=227), wild freshwater (n=227) and wild marine/estuarine species (n=255) of fish in Western Australia by PCR lification at the 18S rRNA locus. Results revealed a low prevalence of Cryptosporidium infection in fish hosts 0.8% (6/709). Four species of Cryptosporidium were identified including C. parvum, C. xiaoi and pig genotype II in whiting (Sillago vittata) and a novel Cryptosporidium spp. in mullets (Mugil cephalus). The identification of zoonotic species of Cryptosporidium in fish indicates that future research to gain a better understanding of the public health impacts is warranted. The detection of the protozoa in fish may also be a good sentinel for environmental contamination or ecosystem health.
Publisher: Elsevier BV
Date: 06-2012
DOI: 10.1016/J.EXPPARA.2012.03.013
Abstract: The woylie or brush-tailed bettong (Bettongia penicillata) is a medium-sized native Australian marsupial that has undergone a dramatic decline in numbers in recent years. Trypanosome parasites have been identified in the woylie but little is known about the prevalence and clinical impact of other haemoprotozoan parasites in these marsupials. In the present study, the occurrence and molecular phylogeny of a piroplasm was studied in woylies from six different sites in Western Australia (WA). Blood s les were screened by PCR at the 18S rRNA locus and 80.4% (123/153) of the blood s les were positive for piroplasm DNA. Sequence and phylogenetic analysis of 12 of these positives identified them as Theileria penicillata, and sequencing of cloned PCR products indicated that no other species of Theileria were present. Infected woylies had a lower body weight but microscopic evaluation of the blood films indicated that T. penicillata did not appear to cause red cell injury or anaemia. Further studies are required to determine the clinical significance of T. penicillata in woylies.
Publisher: Elsevier BV
Date: 07-2016
DOI: 10.1016/J.EXPPARA.2016.04.009
Abstract: An Eimeria species is described from a domestic pigeon (Columba livia domestica). Sporulated oocysts (n = 35) were subspherical, with a smooth bi-layered oocyst wall (1.0 μm thick). Oocysts measured 20.2 × 16.1 (22.0-18.9 × 15.7-18.9) μm, oocyst length/width (L/W) ratio, 1.38. Oocyst residuum and a polar granule were present. The micropyle was absent. Sporocysts are elongate-ovoid, 13.0 × 6.1 (14.5-12.5 × 5.5-7.0) μm, sporocyst L/W ratio, 2.13 (2.0-2.2), sporocyst residuum was present, composed of numerous granules in a spherical or ovoid mass. Each sporocyst contained 2 banana-shaped sporozoites, 12.3 × 3.5 (11.8-13.0 × 3.3-3.6) μm. A spherical-ellipsoid posterior refractile body was found in the sporozoites. A nucleus was located immediately anterior to the posterior refractile body. Molecular analysis was conducted at three loci the 18S and 28S ribosomal RNA genes and the mitochondrial cytochrome oxidase gene (COI). At the 18S locus, the new isolate shared 98.0% genetic similarity with three Isospora isolates from Japan from the domestic pigeon (Columba livia domestica). At the 28S locus, it grouped separately and shared 92.4% and 92.5% genetic similarity with Isospora anthochaerae (KF766053) from a red wattlebird (Anthochaera carunculata) from Australia and an Isospora sp. (MS-2003 - AY283845) from a Himalayan grey-headed bullfinch (Pyrrhula erythaca) respectively. At COI locus, this new isolate was in a separate clade and shared 95.6% and 90.0% similarity respectively with Eimeria tiliquae n. sp. from a shingleback skink in Australia and an Eimeria sp. from a common pheasant (Phasianus colchicus) from America. Based on the morphological data, this isolate is most similar to Eimeria labbeana. As no molecular data for E. labbeana is available and previous morphological data is incomplete, we refer to the current isolate as E. labbeana-like.
Publisher: MDPI AG
Date: 11-05-2020
DOI: 10.3390/MICROORGANISMS8050715
Abstract: Cryptosporidium is a major cause of severe diarrhea-related disease in children in developing countries, but currently no vaccine or effective treatment exists for those who are most at risk of serious illness. This is partly due to the lack of in vitro culturing methods that are able to support the entire Cryptosporidium life cycle, which has led to research in Cryptosporidium biology lagging behind other protozoan parasites. In vivo models such as gnotobiotic piglets are complex, and standard in vitro culturing methods in transformed cell lines, such as HCT-8 cells, have not been able to fully support fertilization occurring in vitro. Additionally, the Cryptosporidium life cycle has also been reported to occur in the absence of host cells. Recently developed bioengineered intestinal models, however, have shown more promising results and are able to reproduce a whole cycle of infectivity in one model system. This review evaluates the recent advances in Cryptosporidium culturing techniques and proposes future directions for research that may build upon these successes.
Publisher: Elsevier BV
Date: 05-2008
DOI: 10.1016/J.EXPPARA.2008.01.010
Abstract: Cryptosporidium hominis, which has an anthroponotic transmission cycle and Cryptosporidium parvum, which is zoonotic, are the primary species of Cryptosporidium that infect humans. The present study identified the species/genotypes and subgenotypes of Cryptosporidium in 7 human and 15 cattle cases of sporadic cryptosporidiosis in rural western NSW during the period from November 2005 to January 2006. The species/genotype of isolates was determined by PCR sequence analysis of the 18S rRNA and C. parvum and C. hominis isolates were subgenotyped by sequence analysis of the GP60 gene. Fourteen of 15 cattle-derived isolates were identified as C. parvum and 1 as a C. bovis/C. parvum mixture. Of the human isolates, 4 were C. parvum and 3 were C. hominis. Two different subgenotypes were identified with the human C. hominis isolates and six different subgenotypes were identified within the C. parvum species from humans and cattle. All four of the C. parvum subtypes found in humans were also found in the cattle, indicating that zoonotic transmission may be an important contributor to sporadic human cases cryptosporidiosis in rural NSW.
Publisher: Elsevier BV
Date: 03-2018
DOI: 10.1016/J.TTBDIS.2018.02.001
Abstract: Piroplasms, including the genera Babesia and Theileria, are intra-erythrocytic protozoa that are generally transmitted by ticks and are the aetiological agents for piroplasmosis in animals, as well as humans, worldwide. In Australia, numerous studies have been conducted on piroplasms in domestic animals however, less is known about these protozoa in ticks from native wildlife. The present study characterised piroplasms in Ixodes australiensis (n = 119) and Amblyomma triguttatum (n = 35) ticks collected from kangaroos in Western Australia (WA). Approximately 7.6% (9/119) (95% CI 2.8-12.2) of the I. australiensis ticks were positive for piroplasms using nested-PCR at the 18S rRNA locus, whereas no piroplasm 18S rDNA was detected in the A. triguttatum ticks. All sequences from I. australiensis ticks were identical. Using a 852 bp multiple nucleotide alignment at the 18S rRNA variable region, sequences shared 97.6%, 94.3%, 93.5% and 93.4% pairwise identity with Theileria fuliginosa, Theileria brachyuri, Theileria penicillata, and a Theileria sp. (K1), derived from a burrowing bettong or boodie (Bettongia lesueur), respectively. Phylogenetic analysis revealed that the Theileria sp. from I. australiensis clustered together in the marsupial-associated Theileria group, with T. fuliginosa as closest sister species. Hence, we conclude that this is the first observation of T. fuliginosa-like species in I. australiensis ticks parasitising kangaroos in WA.
Publisher: Elsevier BV
Date: 2019
DOI: 10.1016/J.IJPARA.2018.07.003
Abstract: Foodborne zoonotic pathogens are a serious public health issue and result in significant global economic losses. Despite their importance to public health, epidemiological data on foodborne diseases including giardiasis caused by the enteric parasite, Giardia duodenalis, are lacking. This parasite is estimated to cause ∼28.2 million cases of diarrhoea each year due to contamination of food, but very few foodborne outbreaks have been documented due to the limitations of current detection as well as surveillance methods. The current method for the recovery of Giardia cysts from food matrices using immunomagnetic separation requires further standardisation and cost reduction before it can be widely used. It also should incorporate downstream molecular procedures for genotyping, and traceback and viability analyses. Foodborne giardiasis can be potentially controlled through improvements in national disease surveillance systems and the establishment of Hazard Analysis and Critical Control Point interventions across the food chain. Studies are needed to assess the true prevalence and public health impact of foodborne giardiasis.
Publisher: Springer Science and Business Media LLC
Date: 19-05-2018
Publisher: Elsevier BV
Date: 10-2007
DOI: 10.1016/J.EXPPARA.2007.03.016
Abstract: Babesia gibsoni is a protozoan parasite of dogs worldwide yet both an effective treatment and a reliable method for detecting subclinical cases of this emerging infection remain elusive. Experimental B. gibsoni infections were established in vivo to investigate the efficacy of combined atovaquone and azithromycin drug therapy and to determine the detection limits of a nested-PCR, IFAT and microscopy during various stages of infection. While atovaquone and azithromycin produced a reduction in parasitaemia, it did not eliminate the parasite and drug resistance appeared to develop in one dog. Polymerase chain reaction was found to be most useful in detecting infection in the pre-acute and acute stages, while IFAT was most reliable during chronic infections. Microscopy is suggested to be only effective for detecting acute stage infections. This study also describes the detection of B. gibsoni in tissue s les during chronic infections for the first time, suggesting possible sequestration of this parasite.
Publisher: Wiley
Date: 02-2001
DOI: 10.1002/1522-2683(200102)22:3<433::AID-ELPS433>3.0.CO;2-K
Publisher: Elsevier BV
Date: 03-2017
DOI: 10.1016/J.VETMIC.2017.01.021
Abstract: Q fever is an infectious disease with a global distribution caused by the intracellular bacterium, Coxiella burnetii, which has been detected in a large number of tick species worldwide, including the brown dog tick, Rhipicephalus sanguineus. Recent reports of a high seroprevalance of C. burnetii in Australian dogs, along with the identification of additional Coxiella species within R. sanguineus ticks, has prompted an investigation into the presence and identification of Coxiella species in R. sanguineus ticks in Australia. Using a combination of C. burnetii species-specific IS1111a transposase gene and Coxiella genus-specific 16S rRNA PCR assays, a Coxiella sp. was identified in 100% (n=199) of R. sanguineus ticks analysed, and C. burnetii was not detected in any R. sanguineus ticks studied. Phylogenetic analysis of the 16S rRNA gene revealed the Coxiella sequences were closely related to Coxiella sp. identified previously in R. sanguineus and R. turanicus ticks overseas. This study illustrates the value of using genus specific PCR assays to detect previously unreported bacterial species. Furthermore, the presence of an additional Coxiella sp. in Australia requires further investigation into its potential for contributing to serological cross-reactions during Q fever testing.
Publisher: Elsevier BV
Date: 09-2011
DOI: 10.1016/J.CIMID.2011.07.002
Abstract: A variety of Bartonella species were detected in two species of ticks and three species of fleas collected from marsupial hosts brush-tailed bettong or woylie (Bettongia penicillata) and western barred bandicoots (Perameles bougainville) and from a rodent host Rattus fuscipes in Western Australia. Bartonella species were detected using nested-PCR of the gltA gene and the 16S-23S ribosomal internal transcribed spacer region (ITS), and species were characterized using DNA sequencing of the 16S rRNA, gltA, rpoB, ftsZ genes and the ITS region. Bartonella rattaustraliani and B. coopersplainsensis were detected in Ixodes spp. ticks and fleas (Stephanocircus pectinipes) respectively collected from rodents. Two novel Bartonella species were detected from marsupials Candidatus Bartonella woyliei n. sp. was detected in both fleas (Pygiopsylla hilli) and ticks (Ixodes australiensis) collected from woylies and Candidatus Bartonella bandicootii n. sp. was detected in fleas (Pygiopsylla tunneyi) collected from western barred bandicoots. Concatenated phylogenetic analysis of all 5 loci clarified the marsupial cluster of Bartonella species in Australia and confirmed the species status of these two Bartonella species in ticks and fleas from woylies and western barred bandicoots, which are classified as threatened species and are vulnerable to extinction.
Publisher: Springer Science and Business Media LLC
Date: 26-06-2018
DOI: 10.1515/AP-2018-0049
Abstract: Cryptosporidium is an important enteric parasite that can contribute large numbers of infectious oocysts to drinking water catchments. As a result of its resistance to disinfectants including chlorine, it has been responsible for numerous waterborne outbreaks of gastroenteritis. Wildlife and livestock play an important role in the transmission of Cryptosporidium in the environment. Studies conducted outside Australia have indicated that camels may also play a role in the transmission of zoonotic species of Cryptosporidium . Despite Australia being home to the world’s largest camel herd, nothing is known about the prevalence and species of Cryptosporidium infecting camels in this country. In the present study, C. parvum was identified by PCR lification and sequencing of a formalin-fixed intestinal tissue specimen from a one-week old dromedary camel ( Camelus dromedarius ). Subtyping analysis at the glycoprotein 60 ( gp60 ) locus identified C. parvum subtype IIaA17G2R1, which is a common zoonotic subtype reported in humans and animals worldwide. Histopathological findings also confirmed the presence of large numbers of variably-sized (1–3 µm in diameter) circular basophilic protozoa – consistent with Cryptosporidium spp.– adherent to the mucosal surface and occasionally free within the lumen. Further analysis of the prevalence and species of Cryptosporidium in camel populations across Australia are essential to better understand their potential for contamination of drinking water catchments.
Publisher: Elsevier BV
Date: 08-2018
Publisher: American Society of Parasitologists
Date: 12-2000
Publisher: American Society for Microbiology
Date: 12-2006
DOI: 10.1128/AEM.01352-06
Abstract: A total of 430 avian-derived fecal specimens were randomly collected from selected Western Australian commercial aviaries, poultry farms, hatcheries, wildlife parks, and the Perth Zoo and screened for the presence of Cryptosporidium by PCR. Of these, 27 Cryptosporidium -positive isolates were detected, characterized, and compared with 11 avian-derived isolates from the Czech Republic at the 18S rRNA and actin gene loci. Sequence and phylogenetic analysis identified four genetically distinct genotypes, avian genotypes I to IV, from various avian hosts. In addition, the host range for Cryptosporidium galli was extended. Cryptosporidium muris and Cryptosporidium andersoni were also identified in a tawny frogmouth and a quail-crested wood partridge, respectively.
Publisher: Elsevier BV
Date: 12-2021
DOI: 10.1016/J.IJPARA.2021.08.007
Abstract: The protozoan parasites Cryptosporidium and Giardia are significant causes of diarrhoea worldwide and are responsible for numerous waterborne and foodborne outbreaks of diseases. Over the last 50 years, the development of improved detection and typing tools has facilitated the expanding range of named species. Currently at least 44 Cryptosporidium spp. and >120 genotypes, and nine Giardia spp., are recognised. Many of these Cryptosporidium genotypes will likely be described as species in the future. The phylogenetic placement of Cryptosporidium at the genus level is still unclear and further research is required to better understand its evolutionary origins. Zoonotic transmission has long been known to play an important role in the epidemiology of cryptosporidiosis and giardiasis, and the development and application of next generation sequencing tools is providing evidence for this. Comparative whole genome sequencing is also providing key information on the genetic mechanisms for host specificity and human infectivity, and will enable One Health management of these zoonotic parasites in the future.
Publisher: Wiley
Date: 06-2001
DOI: 10.1111/J.1550-7408.2001.TB00439.X
Abstract: Cryptosporidium meleagridis is a common cause of cryptosporidiosis in birds. In addition, recent reports have described the parasite as an etiologic agent of cryptosporidiosis in both immunocompetent and immunocompromised humans. Therefore, it is important to genetically characterize isolates of C. meleagridis from different hosts and geographic areas, and to develop molecular tools to differentiate isolates from various hosts or areas. In this study, a total of 11 isolates of Cryptosporidium meleagridis from both human and avian hosts were examined at three genetic loci: the small-subunit rRNA, 60-kDa glycoprotein precursor, and 70-kDa heat shock protein genes. Two genotypes of C. meleagridis were seen at the small-subunit rRNA locus. These differed from each other by the presence or lack of a heterogeneous copy of the gene and an ATT repeat. The 60-kDa glycoprotein precursor gene ided these eleven isolates of C. meleagridis into six genotypes with high sequence ersity between groups. The highest genetic heterogeneity, however, was seen at the 70-kDa heat shock protein locus, and was primarily present at the 3' end of the gene. This heterogeneity separated eight isolates of C. meleagridis into six genotypes. These data could be useful in the development of molecular tools to promote understanding of the transmission of C. meleagridis in humans.
Publisher: Cold Spring Harbor Laboratory
Date: 05-11-2019
DOI: 10.1101/819060
Abstract: Invasive rodent species are known hosts for a erse range of infectious microorganisms and have long been associated with the spread of disease globally. The present study describes molecular evidence for the presence of a Trypanosoma sp. from black rats ( Rattus rattus ) in northern Sydney, Australia. Sequences of the 18S ribosomal RNA (rRNA) locus were obtained in two out of eleven (18%) blood s les with subsequent phylogenetic analysis confirming the identity within the Trypanosoma lewisi clade.
Publisher: American Society for Microbiology
Date: 07-2003
DOI: 10.1128/AEM.69.7.4302-4307.2003
Abstract: Isolates of Cryptosporidium from the Czech Republic were characterized from a variety of different hosts using sequence and phylogenetic analysis of the 18S ribosomal DNA and the heat-shock (HSP-70) gene. Analysis expanded the host range of accepted species and identified several novel genotypes, including horse, Eurasian woodcock, rabbit, and cervid genotypes.
Publisher: American Society of Parasitologists
Date: 04-2003
Publisher: Wiley
Date: 10-2002
DOI: 10.1111/J.1751-0813.2002.TB10961.X
Abstract: Small intraerythrocytic parasites were observed in the blood of three related male American Pit Bull Terriers. Two of the dogs, both less than 1-year-old, were anaemic at the time of initial examination and the third, an adult and sire of the two younger dogs, had a normal haemogram and low parasitaemia. The morphological appearance of the erythrocyte inclusions, analysis of a 450-bp region of the 18S rRNA gene and antibody titres provided evidence that this parasite was Babesia gibsoni, a species not previously reported in Australia.
Publisher: Elsevier BV
Date: 05-2011
DOI: 10.1016/J.VETMIC.2010.12.003
Abstract: Bartonella are fastidious, Gram-negative, aerobic bacilli belonging to the Alphaproteobacteria group. In the last ten years, the discovery of new Bartonella species from a variety of mammalian hosts, arthropod vectors and geographical areas has increased. More than 20 species of Bartonella have been identified, of which approximately thirteen are associated with disease in humans and animals. Recently, four novel species of Bartonella were isolated from mammalian hosts in Australia: Bartonella australis from eastern grey kangaroos (Macropus giganteus) and Bartonella rattaustraliani, Bartonella queenslandensis and Bartonella coopersplainsensis from rodents. Bartonella-like organisms have also been detected from Ixodes tasmani ticks collected from koalas (Phascolarctos cinereus). However, very little is known about Bartonella spp. in other marsupials in Australia. We report the identification of a novel Bartonella species detected from fleas (Acanthopsylla jordani) and ticks (Ixodes antechini) collected from a small carnivorous marsupial, Antechinus flavipes (Mardos or Yellow-footed antechinus) in the southwest of Western Australia. New nested-PCRs targeting the gltA gene and the ribosomal ITS region were developed as part of the present study. DNA sequencing of the 16S rRNA, gltA, ftsZ and rpoB genes and the ribosomal ITS region revealed that this detection is a distinct Bartonella species and is related to B. australis isolated from kangaroos. This is the first report of two different possible arthropod vectors in Australia (ticks and fleas) being infected with the same species of Bartonella. We propose the name Candidatus Bartonella antechini n. sp. for the recently characterized organism.
Publisher: Elsevier BV
Date: 12-2012
Publisher: Microbiology Society
Date: 04-2017
Publisher: American Society for Microbiology
Date: 05-2000
DOI: 10.1128/AEM.66.5.2220-2223.2000
Abstract: Genetic and phylogenetic characterization of Cryptosporidium isolates at two loci (18S rRNA gene and heat shock gene) from both Australian and United States dogs demonstrated that dog-derived Cryptosporidium isolates had a distinct genotype which is conserved across geographic areas. Phylogenetic analysis provided support for the idea that the “dog” genotype is, in fact, a valid species.
Publisher: American Society of Parasitologists
Date: 08-2004
DOI: 10.1645/GE-202R1
Publisher: Cambridge University Press (CUP)
Date: 20-08-2012
DOI: 10.1017/S0031182012001151
Abstract: Cryptosporidium is an important enteric parasite that is transmitted via the fecal-oral route, water and food. Humans, wildlife and domestic livestock all potentially contribute Cryptosporidium to surface waters. Most species of Cryptosporidium are morphologically indistinguishable and can only be identified using molecular tools. Over 24 species have been identified and of these, 7 Cryptosporidium species/genotypes are responsible for most human cryptosporidiosis cases. In Australia, relatively few genotyping studies have been conducted. Six Cryptosporidium species ( C. hominis , C. parvum , C. meleagridis , C. fayeri , C. andersoni and C. bovis) have been identified in humans in Australia. However, little is known about the contribution of animal hosts to human pathogenic strains of Cryptosporidium in drinking water catchments. In this review, we focus on the available genotyping data for native, feral and domestic animals inhabiting drinking water catchments in Australia to provide an improved understanding of the public health implications and to identify key research gaps.
Publisher: American Society for Microbiology
Date: 12-2004
DOI: 10.1128/AEM.70.12.7574-7577.2004
Abstract: Of 471 specimens examined from foxes, raccoons, muskrats, otters, and beavers living in wetlands adjacent to the Chesapeake Bay, 36 were positive for five types of Cryptosporidium , including the C. canis dog and fox genotypes, Cryptosporidium muskrat genotypes I and II, and Cryptosporidium skunk genotype. Thus, fur-bearing mammals in watersheds excreted host-adapted Cryptosporidium oocysts that are not known to be of significant public health importance.
Publisher: Elsevier BV
Date: 04-2017
DOI: 10.1016/J.VETPAR.2017.04.007
Abstract: A molecular survey was conducted to provide baseline information on the prevalence, genetic ersity and potential clinical impacts of blood-borne and enteric protozoans in native wild mammals from the Northern Territory (NT). A total of 209 blood and 167 faecal s les were collected from four target species the northern brown bandicoot (Isoodon macrourus), common brushtail possum (Trichosurus vulpecula), northern quoll (Dasyurus hallucatus) and brush-tailed rabbit-rat (Conilurus penicillatus). Blood s les were screened by PCR at the 18S rRNA gene for trypanosomes, piroplasms and haemogregarines, with faecal s les tested for Cryptosporidium spp. at the 18S rRNA locus, and for Giardia spp. at the glutamate dehydrogenase (gdh) and 18S rRNA loci. The potential clinical impact was investigated by associating clinical, haematological and biochemical parameters with presence or absence of infection. Overall, 22.5% (95% CI: 17.0-28.8%) of the animals tested were positive for haemoprotozoans. Trypanosomes were found in 26.6% (95% CI: 18.7-35.7%) of the bandicoots and were identified as Trypanosoma vegrandis G6, except for one unique genotype, most similar to T. vegrandis G3 (genetic distance=7%). The prevalence of trypanosomes in possums was 23.7% (95% CI: 11.4-40.2%), and the genotypes identified clustered within the T. noyesi clade. The presence of Babesia sp. and Hepatozoon sp. was confirmed in bandicoots only, both at a prevalence of 9.7% (95% CI: 2.7-9.2%). The total prevalence of intestinal protozoan parasites observed was relatively low (3% 95% CI: 1.0-6.9%). No evidence of clinical disease associated with protozoan parasitic infection was observed, however bandicoots positive for Trypanosoma exhibited a significantly lower packed cell volume (PCV) compared to negative bandicoots (p=0.046). To the authors' knowledge, this is the first research conducted in the NT to characterise protozoan parasites in threatened native mammals using both molecular and morphological tools and to assess the potential clinical impacts of these agents. The absence of clear signs of major morbidity in infected animals seems to exclude a direct association between infections with these agents and possible population decline events in northern Australian native mammals. However until the cause(s) of population decline are ascertained for each in idual mammal species, further studies are required. The outcome of the present investigation may be used to inform wildlife conservation and zoonotic disease programs.
Publisher: Elsevier BV
Date: 06-2010
DOI: 10.1016/J.IJPARA.2009.12.001
Abstract: Apart from a single record in a shark, there have been no published studies conducted on Giardia genotypes in fish. The present study investigated the prevalence of Giardia in cultured fingerlings (n=227), wild freshwater (n=227) and wild marine/estuarine species (n=255) of fish in Western Australia by PCR lification at the 18S rRNA, glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi) and beta-giardin (bg) loci. Results revealed a low prevalence of Giardia, 3.8% (27/709), in fish hosts. The zoonotic Giardia species, Giardia duodenalis assemblages A, B as well as G. duodenalis assemblage E and Giardia microti were detected. The identification of zoonotic species of Giardia highlights the public health importance of investigating parasites within fish host species.
Publisher: American Society of Parasitologists
Date: 08-2003
DOI: 10.1645/GE-74RI
Publisher: Elsevier BV
Date: 09-2015
Publisher: Elsevier BV
Date: 06-2015
DOI: 10.1016/J.EXPPARA.2015.03.002
Abstract: A cell-free culture system for Cryptosporidium parvum was analysed using scanning electron microscopy (SEM) to characterise life cycle stages and compare gene expression in cell-free culture and cell culture using HCT-8 cells. Cryptosporidium parvum s les were harvested at 2 h, 8 h, 14 h, 26 h, 50 h, 74 h, 98 h, 122 h and 170 h, chemically fixed and specimens were observed using a Zeiss 1555 scanning electron microscope. The presence of sporozoites, trophozoites and type I merozoites were identified by SEM. Gene expression in cell culture and cell-free culture was studied using reverse transcriptase quantitative PCR (RT-qPCR) of the sporozoite surface antigen protein (cp15), the glycoprotein 900 (gp900), the Cryptosporidium oocyst wall protein (COWP) and 18S ribosomal RNA (rRNA) genes in both cell free and conventional cell culture. In cell culture, cp15 expression peaked at 74 h, gp900 expression peaked at 74 h and 98 h and COWP expression peaked at 50 h. In cell-free culture, CP15 expression peaked at 98 h, gp900 expression peaked at 74 h and COWP expression peaked at 122 h. The present study is the first to compare gene expression of C. parvum in cell culture and cell-free culture and to characterise life cycle stages of C. parvum in cell-free culture using SEM. Findings from this study showed that gene expression patterns in cell culture and cell-free culture were similar but in cell-free culture, gene expression was delayed for CP15 and COWP in cell free culture compared with the cell culture system and was lower. Although three life cycle stageswere conclusively identified, improvements in SEM methodology should lead to the detection of more life cycle stages.
Publisher: Elsevier BV
Date: 10-2014
DOI: 10.1016/J.EXPPARA.2014.06.018
Abstract: The prevalence of Eimeria in sheep in Australia has not been well described, therefore a quantitative PCR (qPCR) was developed, validated and used to study the prevalence and oocyst concentration in lamb faecal s les at three s ling periods (weaning, post-weaning and pre-slaughter) from eight farms across South Australia, New South Wales, Victoria and Western Australia. A total of 3412 faecal s les were collected from approximately 1182 lambs across the 4 states and screened for the presence of Eimeria using this qPCR at the 18S ribosomal RNA (rRNA) locus. A subset of positives was typed by sequence analysis at the 18S locus. The overall prevalence was 18.1% (95% CI 16.8-19.3%) and of the 616 positives, 118 were successfully genotyped. The prevalence of Eimeria was highest in NSW and peaked at 70% during the post-weaning period. The range of oocyst shedding per gram of faeces (g(-1)) at weaning, post-weaning and pre-slaughter overall across all states was 23-2.1×10(7), 23-1.3×10(7) and 23-2.1×10(5), respectively. Median Eimeria shedding g(-1) was higher during post-weaning (1.1×10(3)) and pre-slaughter (1.1×10(3)) than during weaning (206). The following species were identified: Eimeria crandallis, Eimeria ahsata, Eimeria ovinoidalis, Eimeria weybridgensis and Eimeria cylindrica. Of these, E. crandallis and E. ovinoidalis, the most pathogenic species in sheep were responsible for 58.5% of infections typed. This highlights a need for further research to quantify the production impacts of Eimeria in sheep.
Publisher: Mary Ann Liebert Inc
Date: 12-2011
Abstract: Bartonella are arthropod-borne, fastidious, Gram-negative, and aerobic bacilli distributed by fleas, lice, sand flies, and, possibly, ticks. The zoonotic Bartonella species, Bartonella henselae and Bartonella clarridgeiae, which are the causes of cat scratch disease and endocarditis in humans, have been reported from cats, cat fleas, and humans in Australia. However, to date, there has been no report of B. henselae or B. clarridgeiae in Australian wild animals and their ectoparasites. B. henselae and B. clarridgeiae were detected in fleas (Ctenocephalides felis) from red foxes (Vulpes vulpes), an introduced pest animal species in Australia, and only B. clarridgeiae was detected in blood from one red fox. Phylogenetic analysis of the ribosomal intergenic spacer region revealed that the B. henselae detected in the current study were related to B. henselae strain Houston-1, a major pathogenic strain in humans in Australia, and confirmed the genetic distinctness of B. clarridgeiae. The identification and characterization of Bartonella species in red foxes in the Southwest of Western Australia suggests that red foxes may act as reservoirs of infection for animals and humans in this region.
Publisher: Elsevier BV
Date: 12-2015
DOI: 10.1016/J.EXPPARA.2015.08.020
Abstract: A new species, Isospora serinuse n. sp., (Apicomplexa:Eimeriidae) is described from a single domestic canary (Serinus canaria forma domestica) (subspecies S. c. domestica) in Western Australia. Sporulated oocysts of Isospora serinuse n. sp. are spherical or subspherical, 25.5 (24.4-27.0) × 23.5 (22.0-24.8) μm, with a shape index (length/width) of 1.09 and a smooth bilayered oocyst wall, 1.2 μm thick (outer layer 0.9 μm, inner 0.3 μm). A polar granule is present, but a micropyle and oocyst residuum are absent. The sporocysts are lemon-shaped, 18.9 (17.8-20.2) × 11.8 (10.6-13.0) μm, with a shape index of 1.6. Stieda and substieda bodies are present, the Stieda body being a small crescent shape and the substieda being indistinct. Each sporocyst with four vermiform sporozoites arranged head to tail. A sporocyst residuum is present and composed of numerous granules of different sizes that are scattered among the sporozoites. Morphologically, the oocysts of Isospora serinuse n. sp. were different from those of all known valid Isospora spp. Molecular analysis was conducted at 3 loci: the 18S and 28S ribosomal RNA and two separate regions of subunit I of the mitochondrial cytochrome oxidase (COI) gene (designated COIa and COIb). At the 18S locus, Isospora serinuse n. sp. exhibited 97.5% similarity to Isospora sp. Tokyo from a domestic pigeon (Columba livia domestica) in Japan. At the 28S locus, I. serinuse n. sp. exhibited 94.9% similarity to Isospora anthochaerae n. sp. from a red wattlebird (Anthochaera carunculata) in Australia. At the COIa locus, I. serinuse n. sp. exhibited 95.7% similarity to Isospora sospora sp. ex Apodemus flavicollis from a yellow-necked mouse and Isospora gryphoni from an American goldfinch (Carduelis tristis) respectively. At the COIb locus, I. serinuse n. sp. exhibited 96.7% similarity to an Isospora (iSAT4) from a European pied flycatcher (Ficedula hypoleuca). Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora serinuse n. sp. after its host, the domestic canary (S. canaria forma domestica).
Publisher: Elsevier BV
Date: 06-2015
DOI: 10.1016/J.EXPPARA.2015.03.005
Abstract: Eimeria haematodi was first described in 1977 from the rainbow lorikeet (Trichoglossus haematodus) in Papua New Guinea. In the present study, we re-describe this coccidian species morphologically and molecularly from a rainbow lorikeet bird in Western Australia (WA). The oocysts were ovoid to slightly piriform and measured 28.5-37.8 by 25.8-33.0 µm (33.3 by 28.1 µm). Oocyst wall was approximately 1.5 µm thick and bilayered. Micropyle (5-7 µm) and oocyst residuum (8.0-10.0 µm) present polar granule was absent. Sporocysts ellipsoidal, 11.8-13.6 by 8.0-9.6 µm (12.2 by 8.3 µm), with thin convex Stieda body and granular sporocyst residuum (4.0-5.0 µm). Molecular characterization of E. haematodi was conducted at 18S ribosomal RNA and the mitochondrial cytochrome oxidase gene (COI) loci. At the 18S ribosomal RNA locus, E. haematodi shared 98.1% genetic similarity to E. alabamensis from cattle in New South Wales, Australia. At COI locus, E. haematodi was closest (92.3% similarity) to E. praecox from domestic chickens (Gallus gallus domesticus) from Canada and China.
Publisher: Springer Science and Business Media LLC
Date: 13-01-2016
DOI: 10.1007/S00436-016-4901-0
Abstract: A molecular epidemiological survey of Cryptosporidium from water buffalo (Bubalus bubalis) in the Northern Territory in Australia was conducted. Fecal s les were collected from adult farmed (n = 50) and wild buffalo (n = 50) and screened using an 18S quantitative PCR (qPCR). Positives were typed by sequence analysis of 18S nested PCR products. The qPCR prevalence of Cryptosporidium species in farmed and wild buffalo was 30 and 12 %, respectively. Sequence analysis identified two species: C. parvum and C. bovis, with C. parvum accounting for ~80 % of positives typed from the farmed buffalo fecal s les compared to 50 % for wild buffalo. Subtyping at the 60 kDa glycoprotein (gp60) locus identified C. parvum subtypes IIdA19G1 (n = 4) and IIdA15G1 (n = 1) in the farmed buffalo and IIaA18G3R1 (n = 2) in the wild buffalo. The presence of C. parvum, which commonly infects humans, suggests that water buffaloes may contribute to contamination of rivers and waterways with human infectious Cryptosporidium oocysts, and further research on the epidemiology of Cryptosporidium in buffalo populations in Australia is required.
Publisher: Informa UK Limited
Date: 03-07-2021
Publisher: Wiley
Date: 09-1999
Publisher: Elsevier BV
Date: 09-2003
DOI: 10.1016/S0020-7519(03)00137-1
Abstract: A longitudinal survey of gastro-intestinal parasites was conducted over a 3-year period in remote communities in the north-west of Western Australia where, based on diagnosis by microscopy of faecal s les, Rodentolepis (=Hymenolepis) nana was found to be the most common enteric parasite. In the present study, using molecular tools, we describe the unexpected discovery, of a mixed infection with a second hymenolepidid species, Rodentolepis (=Hymenolepis) microstoma in four of the surveyed in iduals. In the absence of any reliable earlier reports we believe this is to be the first instance of the detection of R. microstoma from human hosts. The development of a diagnostic restriction fragment polymorphism has enabled the study of R. microstoma in human populations and will greatly facilitate a more thorough understanding of the epidemiology of this parasite in the future.
Publisher: Cambridge University Press (CUP)
Date: 07-1998
DOI: 10.1017/S0031182098002765
Abstract: A 298 bp region of the Cryptosporidium parvum 18S rDNA and a 390 bp region of the acetyl-CoA synthetase gene were sequenced for a range of human and animal isolates of Cryptosporidium from different geographical areas. A distinct genotype is common to isolates from cattle, sheep and goats and also an alpaca from Peru and is referred to here as the ‘calf’-derived Cryptosporidium genotype. Another genotype of ‘human’-derived isolates also appears to be conserved amongst human isolates although humans are also susceptible to infection with the ‘calf’ Cryptosporidium genotype. Mice and pigs carry genetically distinct genotypes of Cryptosporidium . Three snake isolates were also analysed, 2 of which exhibited C. muris genotypes and the third snake isolate carried a distinct ‘mouse’ genotype.
Publisher: Ovid Technologies (Wolters Kluwer Health)
Date: 10-2004
DOI: 10.1097/00001432-200410000-00014
Abstract: Molecular tools have been developed to detect and differentiate Cryptosporidium at the species/genotype and subtype levels. These tools have been increasingly used in the characterization of the transmission of Cryptosporidium spp. This review addresses the most recent developments in molecular epidemiology of cryptosporidiosis. The recent development of subtyping tools has led to better understanding of the population genetics and transmission of Cryptosporidium in humans. The population structure of C. parvum and C. hominis is apparently more complicated than previously suggested, with the likely existence of both clonal and panmictic populations. Thus, the transmission of C. parvum (genotype II) in humans is shown to be different in different areas, with zoonotic transmission important in certain places and anthroponotic transmission in others. The use of molecular tools has also led to the identification of geographic and temporal differences in the transmission of C. parvum and C. hominis, and better appreciation of the public health importance of other Cryptosporidium species/genotypes and the frequency of infections with mixed genotypes or subtypes. Factors involved in the transmission of human cryptosporidiosis are difficult to examine using conventional methods. The use of molecular tools has been helpful in the assessment of the zoonotic potential of various Cryptosporidium spp. and sources of human infections, and has started to play a significant role in the characterization of transmission dynamic in endemic and epidemic areas.
Publisher: Cambridge University Press (CUP)
Date: 11-1999
DOI: 10.1017/S0031182099004151
Abstract: In this chapter, the contribution of molecular tools in understanding the aetiology and ecology of infectious diseases is examined in the context of molecular epidemiology (ME). ME is seen as providing the ‘tools’, both laboratory and analytical, which have predictive significance in epidemiological investigations of the causation of disease. A ersity of questions can be addressed with these tools which can conveniently be viewed as particular regions of DNA and grouped according to the different hierarchical levels of specificity by which infectious agents can be characterized. These groupings and the applications of the different molecular tools are described, and consideration given to the most appropriate methods of analysing data from ME investigations.
Publisher: Springer Science and Business Media LLC
Date: 30-10-2019
DOI: 10.1186/S13071-019-3759-2
Abstract: Apicomplexan parasites of the genus Cryptosporidium infect a wide range of animal species as well as humans. Cryptosporidium spp. can cause life threatening diarrhea especially in young animals, children, immunocompromised patients and malnourished in iduals. Asymptomatic cryptosporidial infections in animals can also occur, making these animals potential reservoirs of infection. In the present study, a molecular survey of Cryptosporidium spp. in ruminants that were slaughtered for human consumption in Yazd Province, located in central Iran was conducted. Faeces were collected per-rectum from 484 animals including 192 cattle, 192 sheep and 100 goats. DNA was extracted from all s les and screened for Cryptosporidium by PCR lification of the 18S rRNA gene. Positives were Sanger sequenced and further subtyped by sequence analysis of the 60 kDa glycoprotein ( gp60 ) locus. In total, Cryptosporidium spp. were detected in 22 animals: C. andersoni and C. bovis in seven and two cattle faecal s les, respectively, C. ubiquitum in five sheep, and C. xiaoi in six sheep and two goat s les, respectively. To our knowledge, this study provides for the first time, molecular information concerning Cryptosporidium species infecting goats in Iran, and is also the first report of C. ubiquitum and C. xiaoi from ruminants in Iran. The presence of potentially zoonotic species of Cryptosporidium in ruminants in this region may suggest that livestock could potentially contribute to human cryptosporidiosis, in particular among farmers and slaughterhouse workers, in the area. Further molecular studies on local human populations are required to more accurately understand the epidemiology and transmission dynamics of Cryptosporidium spp. in this region.
Publisher: Wiley
Date: 09-1999
Publisher: Elsevier BV
Date: 09-2021
Publisher: Elsevier BV
Date: 09-2008
DOI: 10.1016/J.EXPPARA.2008.05.002
Abstract: A total of 421 fecal s les from a variety of captive and wild marsupial hosts in Western Australia, Victoria and South Australia were screened for the presence of Giardia species/genotypes using PCR and sequence analysis of a fragment of the 18S rRNA gene. Giardia spp. were identified in 13.4% (28/209) of s les from captive marsupials and 13.7% (29/212) of s les from wild marsupials. Sequence analysis at the 18S locus identified the zoonotic Giardia duodenalis Genotypes A and B in both captive and wild marsupials. Eight isolates were typed as genotype B3 and B4 at the gdh locus, although 7/8 were typed as genotype A at the 18S rRNA locus. The possible reasons for this discordance are discussed. This is the first report of genotype B and only the second report of genotype A in marsupials. As some of the genotype B isolates were identical to human-derived Giardia gdh sequences, these results suggest that marsupials in catchments may pose a public health risk and therefore warrant further investigation.
Publisher: Frontiers Media SA
Date: 10-01-2022
DOI: 10.3389/FMICB.2021.810142
Abstract: Animal farming has intensified significantly in recent decades, with the emergence of concentrated animal feeding operations (CAFOs) in industrialized nations. The congregation of susceptible animals in CAFOs can lead to heavy environmental contamination with pathogens, promoting the emergence of hyper-transmissible, and virulent pathogens. As a result, CAFOs have been associated with emergence of highly pathogenic avian influenza viruses, hepatitis E virus, Escherichia coli O157:H7, Streptococcus suis , livestock-associated methicillin-resistant Staphylococcus aureus , and Cryptosporidium parvum in farm animals. This has led to increased transmission of zoonotic pathogens in humans and changes in disease patterns in general communities. They are exemplified by the common occurrence of outbreaks of illnesses through direct and indirect contact with farm animals, and wide occurrence of similar serotypes or subtypes in both humans and farm animals in industrialized nations. Therefore, control measures should be developed to slow down the dispersal of zoonotic pathogens associated with CAFOs and prevent the emergence of new pathogens of epidemic and pandemic potential.
Publisher: Elsevier BV
Date: 09-2020
Publisher: Cambridge University Press (CUP)
Date: 14-08-2017
DOI: 10.1017/S0031182017001214
Abstract: Currently, the systematics, biology and epidemiology of piscine Cryptosporidium species are poorly understood. Here, we compared Sanger ‒ and next-generation ‒ sequencing (NGS), of piscine Cryptosporidium , at the 18S rRNA and actin genes. The hosts comprised 11 ornamental fish species, spanning four orders and eight families. The objectives were: to (i) confirm the rich genetic ersity of the parasite and the high frequency of mixed infections and (ii) explore the potential of NGS in the presence of complex genetic mixtures. By Sanger sequencing, four main genotypes were obtained at the actin locus, while for the 18S locus, seven genotypes were identified. At both loci, NGS revealed frequent mixed infections, consisting of one highly dominant variant plus substantially rarer genotypes. Both sequencing methods detected novel Cryptosporidium genotypes at both loci, including a novel and highly abundant actin genotype that was identified by both Sanger sequencing and NGS. Importantly, this genotype accounted for 68·9% of all NGS reads from all s les (249 585/362 372). The present study confirms that aquarium fish can harbour a large and unexplored Cryptosporidium genetic ersity. Although commonly used in molecular parasitology studies, nested PCR prevents quantitative comparisons and thwarts the advantages of NGS, when this latter approach is used to investigate multiple infections.
Publisher: Elsevier BV
Date: 03-2014
DOI: 10.1016/J.EXPPARA.2014.01.007
Abstract: The identification and characterisation of novel Eimeria species has largely been based on sporulated oocyst and sporocyst morphology, the host species and the geographical range. Variation in the size and shape of Eimeria oocysts across their host range however, make the identification and characterisation of novel species using traditional methodologies alone problematic. The use of molecular markers and phylogenetic analysis has greatly advanced our ability to characterise Eimeria species and has recently been applied to understand evolutionary relationships among Eimeria species from Australian marsupials. In the present study, Eimeria species isolated from quokkas (Setonix brachyurus) captured from Two Peoples Bay, Bald Island and Rottnest Island, Western Australia, were morphologically identified as Eimeria quokka and Eimeria setonicis. Both Eimeria species were identified as being polymorphic in nature with regards to sporulated oocyst and sporocyst morphometrics. Phylogenetic analysis using 18S rRNA and COI (cytochrome c oxidase subunit 1) genes, grouped E. quokka and E. setonicis within the Eimeria marsupial clade together with Eimeria trichosuri from brushtail possums, Eimeria macropodis from tammar wallabies (Macropus eugenii) and several unidentified macropod Eimeria species from western grey kangaroos (Macropus fuliginosus). This study is the first to characterise E. quokka and E. setonicis by molecular analysis, enabling more extensive resolution of evolutionary relationships among marsupial-derived Eimeria species.
Publisher: Elsevier BV
Date: 04-2016
Publisher: Springer Science and Business Media LLC
Date: 12-08-2020
Publisher: Elsevier BV
Date: 08-2016
DOI: 10.1016/J.VETPAR.2016.07.025
Abstract: Dientamoeba fragilis is a potentially pathogenic, enteric, protozoan parasite with a worldwide distribution. While clinical case reports and prevalence studies appear regularly in the scientific literature, little attention has been paid to this parasite's biology, life cycle, host range, and possible transmission routes. Overall, these aspects of Dientamoeba biology remain poorly understood at best. In this study, a total of 420 animal s les, collected from Australia, were surveyed for the presence of Dientamoeba fragilis using PCR. Several PCR assays were evaluated for sensitivity and specificity. Two previously published PCR methods demonstrated cross reactivity with other trichomonads commonly found in animal s les. Only one assay exhibited excellent specificity. Using this assay D. fragilis was detected from one dog and one cat s le. This is the first report of D. fragilis from these animals and highlights the role companion animals may play in D. fragilis transmission. This study demonstrated that some published D. fragilis molecular assays cross react with other closely related trichomonads and consequently are not suitable for animal prevalence studies.
Publisher: Elsevier BV
Date: 09-1998
Publisher: Springer Science and Business Media LLC
Date: 10-05-2016
Publisher: Elsevier BV
Date: 12-2009
DOI: 10.1016/J.MOLBIOPARA.2009.07.003
Abstract: Cryptosporidium parvum is a protozoan parasite that infects a variety of mammals. The parasite has been shown to harbor a dsRNA virus (CPV) and in the present study, we have developed a CPV transient transfection system for this parasite by using green fluorescent protein (GFP) to replace the partial gene encoding region of the larger dsRNA (CPV-L) and the smaller dsRNA (CPV-S) virus. Two viral RNA-mediated transfection vectors: pCPVL-GFP and pCPVS-GFP were successfully constructed and both in vitro transcripts were electroporated into oocysts and sporozoites. Transient expression of GFP was detected in C. parvum oocysts and excysted sporozoites by fluorescence microscopy and by RT-PCR detection of GFP mRNA and antisense RNA in transfected C. parvum oocysts. Our study provides a new approach for studying gene expression and regulation in C. parvum and will hopefully lead to the construction of a stable CPV transfection system in the future.
Publisher: Elsevier BV
Date: 12-2011
DOI: 10.1016/J.VETPAR.2011.07.009
Abstract: In the present study, the occurrence and molecular phylogeny of trypanosome parasites were studied in both wild and captive marsupials from Western Australia and Queensland. Blood s les were screened by PCR at the 18S rDNA locus, and the glycosomal glyceraldehyde phosphate dehydrogenase gene. Overall, 5.3% of the blood s les were positive at the 18S rDNA locus. All positives belonged to wild-captured Western Australian in iduals, where trypanosome-specific DNA was detected in 9.8% of the screened s les from wild marsupials, in common brushtail possums, and woylies. The detection rate of trypanosome DNA in these two host species was 12.5% and 20%, respectively. Phylogenetic analyses based on two loci, indicated that the possum-derived trypanosome isolates were genetically distinct, and most closely related to the Australian marsupial trypanosomes H25 from a kangaroo, and BRA2 from a bush rat. This is the first study to genetically characterise trypanosome isolates from possums. The analysis of the woylie-derived isolates demonstrated that this marsupial host can harbour multiple genotypes within the same geographical location and furthermore multiple genotypes within the same host, indicative of mixed infections. All the woylie-derived genotypes grouped with trypanosomes found in Australian marsupials, suggesting that they are more likely to belong to an endemic or Australasian trypanosome species. This is the first study to genetically characterise trypanosome isolates from possums (Trichosurus vulpecula). Although the clinical significance of these infections is currently unknown, the identification of these novel sequences may support future investigations on transmission, threats to endangered wildlife, and evolutionary history of the genus Trypanosoma.
Publisher: Elsevier BV
Date: 2010
DOI: 10.1016/J.EXPPARA.2009.02.002
Abstract: Whilst considerable information is available for avian cryptosporidiosis, scant information is available for Cryptosporidium infections in fish and hibians. The present review details recent studies in avian cryptosporidiosis and our current knowledge of piscine and hibian infections.
Publisher: Elsevier BV
Date: 03-2008
DOI: 10.1016/J.EXPPARA.2007.08.019
Abstract: The present study was undertaken to analyse the capability of HIV-1 derived TAT protein transduction domain (PTD) fused with Green Fluorescent Protein (TAT-GFP) as a delivery vehicle into a range of protozoan parasites. Successful transduction of native purified TAT-GFP was observed by fluorescent microscopy in Cryptosporidium parvum, Giardia duodenalis, and Neospora caninum. The ability to transduce peptides and other cargo into protozoan parasites, will greatly assist in the delivery of future peptide-based drugs and target validation peptides.
Location: Singapore
Location: United States of America
No related grants have been discovered for Una Ryan.