ORCID Profile
0000-0003-2563-2015
Current Organisation
Murdoch University
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Publisher: Elsevier BV
Date: 11-2016
DOI: 10.1016/J.EXPPARA.2016.09.012
Abstract: Parasites of wildlife inhabiting urbanised and peri-urban environments are of interest regarding wildlife population health, and also veterinary public health in the case of parasites that can also infect humans and domestic animals. This study aimed to: identify, and estimate the prevalence of, species of Eimeria parasitic in quenda (Isoodon obesulus) in the greater Perth region, Western Australia 2) morphologically describe and genetically characterise a novel observed species of Eimeria as E. angustus and 3) genetically characterise E. kanyana. Eimeria spp. prevalence was 76.1% (95% CI 64.9-84.5%), and four putative species of Eimeria were identified. Eimeria kanyana was identified infecting quenda for the first time, with a prevalence of 54.9% (43.4-66.0%). Eimeria quenda was less prevalent, at 7.0% (3.1-15.5%). The novel species E. angustus was present in 45.1% of s led quenda (34.0-56.6%). A second novel morphotype of Eimeria was present in 2.8% of s led quenda (0.9-9.7%). Mixed Eimeria spp. infections were present in 21/71 quenda (29.6%, 95% CI 20.2-41.1%). Molecular phylogenetic analyses of E. kanyana and E. angustus were conducted at the 18S rRNA and mitochondrial cytochrome oxidase loci. At both loci, two isolates identified as E. kanyana grouped in a phylogenetic clade with E. trichosuri. Five isolates identified as the novel E. angustus were most closely related to E. tropidura at the 18S locus. At the COI locus, no sequence data were available for E. tropidura isolates of E. angustus grouped with E. sciurorum.
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: Wiley
Date: 09-2023
DOI: 10.1002/ECE3.10505
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: 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: 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: 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: 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: 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: 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: 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: 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: 09-08-2016
DOI: 10.1038/SREP30692
Abstract: The wheat avenin-like proteins (ALP) are considered atypical gluten constituents and have shown positive effects on dough properties revealed using a transgenic approach. However, to date the genetic architecture of ALP genes is unclear, making it impossible to be utilized in wheat breeding. In the current study, three genes of type-b ALPs were identified and mapped to chromosomes 7AS, 4AL and 7DS. The coding gene sequence of both TaALP-7A and TaALP-7D was 855 bp long, encoding two identical homologous 284 amino acid long proteins. TaALP-4A was 858 bp long, encoding a 285 amino acid protein variant. Three alleles were identified for TaALP-7A and four for TaALP-4A. TaALP-7A alleles were of two types: type-1, which includes TaALP-7A1 and TaALP-7A2 , encodes mature proteins, while type-2, represented by TaALP-7A3 , contains a stop codon in the coding region and thus does not encode a mature protein. Dough quality testing of 102 wheat cultivars established a highly significant association of the type-1 TaALP-7A allele with better wheat processing quality. This allelic effects were confirmed among a range of commercial wheat cultivars. Our research makes the ALP be the first of such genetic variation source that can be readily utilized in wheat breeding.
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: 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: 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: 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: 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: 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: CSIRO Publishing
Date: 2018
DOI: 10.1071/CP18273
Abstract: The wheat NAM-B1 and NAM-A1 genes are positively associated with grain protein content (GPC) in wheat. We conducted molecular characterisation of the NAM-1 genes in 51 Australian wheat varieties (Triticum aestivum L.), with the aim of improving GPC and nitrogen-usage efficiency in Australian wheat. In summary, the wild type NAM-B1 gene, which originated from Israel, was identified in two Australian wheat varieties. Five varieties contained a deletion allele, whereas the majority (43) harboured a non-functional NAM-B1 allele and one variety contained both functional and non-functional alleles. Twenty-six Australian wheat varieties contained the NAM-A1a haplotype, which was similar to its well-characterised homoeolog NAM-B1 wild type and associated with high GPC. The NAM-D1 gene in the 51 wheat varieties was also characterised, and no gene variation in the exon regions was noted only two single-nucleotide polymorphisms in introns 1 and 2 were found among the 51 varieties.
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: 05-2017
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: 02-2021
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: 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 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: 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: Springer Science and Business Media LLC
Date: 24-09-2018
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: 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: 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: 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: 2021
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: 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: 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: 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: 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: 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: 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: 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: Springer Science and Business Media LLC
Date: 22-07-2019
DOI: 10.1007/S11230-019-09870-Y
Abstract: Faecal s les (n = 1,093) collected from the woylie Bettongia penicillata Gray, in south-western Australia were examined for the presence of coccidian parasites. Eimeria sp. oöcysts were detected in 15.2% of s les. Faecal s les obtained from the eastern bettong Bettongia gaimardi (Desmarest) (n = 4) and long-nosed potoroo Potorous tridactylus (Kerr) (n = 12) in Tasmania, were also screened for the presence of Eimeria spp. (prevalence 50% and 41.7%, respectively). Morphological and genetic comparison with other known species of Eimeria indicates that the material identified in woylies is novel. This study aimed to (i) morphologically describe and genetically characterise Eimeria woyliei n. sp. found in woylies and (ii) genetically characterise Eimeria gaimardi Barker, O'Callaghan & Beveridge, 1988, Eimeria potoroi Barker, O'Callaghan & Beveridge, 1988, and Eimeria mundayi Barker, O'Callaghan & Beveridge, 1988, from other potoroid marsupials. Molecular phylogenetic analyses conducted at the 18S rDNA and mitochondrial cytochrome c oxidase subunit 1 (cox1) loci revealed that E. woyliei n. sp. was most closely related to Eimeria setonicis Barker, O'Callaghan & Beveridge, 1988, at the 18S rDNA locus, and Eimeria trichosuri O'Callaghan & O'Donoghue, 2001, at the cox1 locus. Eimeria woyliei n. sp. is the sixth species of Eimeria to be formally described from potoroid marsupials.
Publisher: Cold Spring Harbor Laboratory
Date: 03-09-2018
DOI: 10.1101/406694
Abstract: Wheat Avenin-like proteins (TaALP) are atypical storage proteins belonging to the Prolamin superfamily. Previous studies on ALPs have focused on the proteins’ positive effects on dough strength, whilst no correlation has been made between TaALPs and the plant immune system. Here, we performed genome-wide characterization of ALP encoding genes in bread wheat. In silico analyses indicated the presence of critical peptides in TaALPs that are active in the plant immune system. Pathogenesis-related nucleotide motifs were also identified in the putative promoter regions of TaALP encoding genes. RT-PCR was performed on TaALP and previously characterised pathogenesis resistance genes in developing wheat caryopses under control and Fusarium graminearum infection conditions. The results showed that TaALP and NMT genes were upregulated upon F. graminearum inoculation. mRNA insitu hybridization showed that TaALP genes were expressed in the embryo, aleurone and sub-aleurone layer cells. Seven TaALP genes were cloned for the expression of recombinant proteins in Escherichia coli , which displayed significant inhibitory function on F. graminearum under anti-fungal tests. In addition, FHB index association analyses showed that allelic variations of two ALP genes on chromosome 7A were significantly correlated with FHB symptoms. Over-expression of an ALP gene on chromosome 7A showed an enhanced resistance to FHB. Yeast two Hybridization results revealed that ALPs have potential proteases inhibiting effect on metacaspases and beta-glucosidases. A vital infection process related pathogen protein, F. graminearum Beta-glucosidase was found to interact with ALPs. Our study is the first to report a class of wheat storage protein or gluten protein with biochemical functions. Due to its abundance in the grain and the important multi-functions, the results obtained in the current study are expected to have a significant impact on wheat research and industry.
Publisher: Elsevier BV
Date: 2016
Publisher: Elsevier BV
Date: 03-2016
Publisher: Elsevier BV
Date: 03-2016
Publisher: Springer Science and Business Media LLC
Date: 21-05-2016
Publisher: Elsevier BV
Date: 03-2016
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: 2018
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: 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: Authorea, Inc.
Date: 12-06-2023
DOI: 10.22541/AU.168654984.48636760/V1
Abstract: A new coccidian species, Isospora elliotae n. sp., from the Australian magpie Gymnorhina tibicen (Latham, 1801) in Western Australia is described and characterised morphologically and molecularly. Microscopic analysis of a faecal s le identified subspheroidal oöcysts (n = 20), 20–22 × 18–20 (20.7 × 18.7) length/width (L/W) ratio 1.05–1.14 (1.10). Wall bi-layered, 1.0–1.3 (1.2) thick, outer layer smooth, c.2/3 of total thickness. Micropyle and oocyst residuum absent, but usually two polar granules are present. Sporocysts (n = 28) ovoidal, 12–13 × 9–11 (12.6 × 9.7) L/W ratio 1.22–1.35 (1.30). Stieda body present, flattened to half-moon-shaped, c. 0.5 deep × 2.0 wide sub-Stieda indistinct or barely discernible, c. 1.0 deep × 2.5 wide para-Stieda body absent sporocyst residuum present, composed of granules dispersed among the sporozoites. Sporozoites vermiform, with anterior and posterior refractile bodies and nucleus. Segments of three gene loci (18S rRNA, 28S rRNA and COI) were sequenced and I. elliotae n. sp. exhibited 99.8% genetic similarity to Isospora sp. MAH-2013a (KF648870) followed by 99.7% genetic similarity to Isospora neochmiae Yang, Brice & Ryan, 2016 (KT224380) at the 18S rRNA gene locus. It shared 97.0% genetic similarity with an unnamed Isospora sp. (AY283852) at the 28S rRNA gene locus and it also shared the highest genetic similarity of 99.8% with the unnamed Isospora sp. from an American crow (OL999120) at the COI gene locus. Based on morphological and molecular data, this isolate is a new species named as I. elliotae n. sp.
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: 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: 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: 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: 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: 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: 09-2015
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: Public Library of Science (PLoS)
Date: 14-12-2016
Publisher: Wildlife Disease Association
Date: 10-2011
DOI: 10.7589/0090-3558-47.4.821
Abstract: We investigated the role of the western grey kangaroo (Macropus fuliginosus) in the maintenance and transmission of Coxiella burnetii in Western Australia. Sera from 1,017 kangaroos were tested using an indirect enzyme-linked immunosorbent assay (ELISA) for the presence of C. burnetii antibodies. The overall antibody prevalence across 12 locations throughout mid- to southwestern Western Australia was 24.1% (95% CI: 21.6-26.8). Feces from 990 of the same animals were tested using PCR to identify active shedding of C. burnetii in excreta. Coxiella burnetii DNA was detected in 4.1% (95% CI: 3.1-5.6) of s les. Our results suggest that kangaroos are reservoirs for C. burnetii in Western Australia and may contribute to transmission of the organism to domestic livestock and humans.
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 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: 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: 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: 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: 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: 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: 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: 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: 05-2007
DOI: 10.1016/J.JBIOTEC.2007.01.008
Abstract: We are investigating the use of single chain antibody fragments (scFv) in eye drops for diagnosis and treatment of eye diseases. For ocular use, recombinant proteins must be free of bacterial endotoxin that causes inflammation in the eye. We required a means of generating high yields of scFvs with little endotoxin contamination. Using microprojectile bombardment we produced transgenic lines of the commercial wheat variety, Westonia, that express two scFvs that bind to CD4 or CD28 on the surface of rat thymocytes. A high level of expression of active scFv in the range 50-180 microg/g was measured by quantitative flow cytometry in crude extracts made from mature seeds. The levels of expression were stable over four generations of transgenic plants and mature seeds were stored for one year with little loss of scFv activity. Substantial purification of scFv was achieved by immobilised metal affinity chromatography. Compared to bacterial extracts, crude transgenic seed extracts contained only a small amount of endotoxin (150 EU/ml) that will be easily removed by purification. The transgenic wheat lines express functional scFv at levels comparable to production in bacteria and promise to be superior to bacteria for production of scFv pharmaceuticals for ocular use.
Publisher: Proceedings of the National Academy of Sciences
Date: 10-12-2018
Abstract: Fifteen full-length wheat grain avenin-like protein coding genes ( TaALP ) were identified on chromosome arms 7AS, 4AL, and 7DS of bread wheat with each containing five genes. Besides the a- and b-type ALPs, a c type was identified in the current paper. Both a and b types have two subunits, named x and y types. The five genes on each of the three chromosome arms consisted of two x-type genes, two y-type genes, and one c-type gene. The a-type genes were typically of 520 bp in length, whereas the b types were of 850 bp in length, and the c type was of 470 bp in length. The ALP gene transcript levels were significantly up-regulated in Blumeria graminis f. sp. tritici (Bgt) -infected wheat grain caryopsis at early grain filling. Wild emmer wheat [(WEW), Triticum dicoccoides ] populations were focused on in our paper to identify allelic variations of ALP genes and to study the influence of natural selection on certain alleles. Consequently, 25 alleles were identified for TdALP-bx-7AS , 13 alleles were identified for TdALP-ax-7AS , 7 alleles were identified for TdALP-ay-7AS , and 4 alleles were identified for TdALP-ax-4AL . Correlation studies on TdALP gene ersity and ecological stresses suggested that environmental factors contribute to the ALP polymorphism formation in WEW. Many allelic variants of ALPs in the endosperm of WEW are not present in bread wheat and therefore could be utilized in breeding bread wheat varieties for better quality and elite plant defense characteristics.
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: 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: 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: Royal Zoological Society of New South Wales
Date: 2016
DOI: 10.7882/AZ.2015.030
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: Public Library of Science (PLoS)
Date: 15-10-2018
Publisher: Wiley
Date: 23-10-2018
DOI: 10.1111/TPJ.14096
Abstract: In wheat (Triticum aestivum) grain yield and grain protein content are negatively correlated, making the simultaneous increase of the two traits challenging. Apart from genetic approaches, modification of nitrogen fertilization offers a feasible option to achieve this aim. In this study, a range of traits related to nitrogen-use efficiency in six Australian bread wheat varieties were investigated under different nitrogen treatments using 3-year multisite field trials. Changes in the in idual storage protein composition were detected by high-performance liquid chromatography. Our results indicated that wheat grain yield and grain protein content reacted similarly to nitrogen availability, with grain yield being slightly more sensitive than grain protein content, and that genotype is a vital determinant of grain protein yield. Measurement of the glutamine synthetase activity of flag leaves and developing grains revealed that high nitrogen availability prompted the participation of glutamine in biological processes. In addition, a more significant accumulation of gluten macropolymer was observed under the high-nitrogen treatment from 21 days post-anthesis, and the underlying mechanism was elucidated by a comparative proteomics study. A yeast two-hybrid experiment confirmed this mechanism. The results of this study revealed that peptidyl-prolyl cis-trans isomerase (PPIase) was SUMOylated with the assistance of small ubiquitin-related modifier 1 and that high nitrogen availability facilitated this connection for the subsequent protein polymerization. Additionally, luminal-binding protein 2 in the endoplasmic reticulum played a similar role to PPIase in the aggregation of protein under high-nitrogen conditions.
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: 04-2009
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: Springer Science and Business Media LLC
Date: 11-08-2020
Publisher: Elsevier BV
Date: 04-2016
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: 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: 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: Springer Science and Business Media LLC
Date: 21-11-2020
DOI: 10.1007/S00436-019-06546-W
Abstract: A new Caryospora-like isolate is described from a magpie-lark (Grallina cyanoleuca) in Western Australia. Sporulated oocysts of the Caryospora-like isolate (n = 35) are subspherical with a shape index of 1.13 ((21.5 (19.7-23.6) × 19.0 (18.1-19.8) μm). The bilayered oocyst wall is smooth. Micropyle, polar granule and oocyst residuum are absent. The sporocyst is ellipsoidal, 18.9 (17.2-20.8) × 12.3 (11.9-12.8) μm, with a shape index (length/width) of 1.54. The sporocyst wall is bilayered. Stieda and substieda bodies are present, the Stieda body is small and flattened and the substieda is trapezoidal. Sporocyst with eight sporozoites arranged head to tail. The sporozoites are vermiform, 18.9 (17.2-20.8) × 12.3 (11.9-12.8) μm and have striations at the anterior end. Each sporozoite has both anterior and posterior refractile bodies. A sporocyst residuum is present. Molecular characterization of the isolated Caryospora-like oocysts was conducted at the 18S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) loci. At the 18S rRNA locus, the Caryospora-like isolate exhibited 88.8% to 96.5% similarity with other Caryospora spp. from different hosts. At the COI locus, it showed 91.5% similarity to Caryospora cf. bigenetica JB-2013 (KF859856) from the rattlesnake, Sistrurus catenatus.
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: 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).
No related grants have been discovered for Rongchang Yang.