ORCID Profile
0000-0002-1105-5184
Current Organisations
University of Sydney
,
Murdoch University
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Publisher: Elsevier BV
Date: 10-1995
Publisher: Elsevier BV
Date: 10-2014
DOI: 10.1016/J.EXPPARA.2014.07.004
Abstract: Little is known about the prevalence and pathogenesis of trypanosomes in Australian monotremes, and few genetic characterisation studies have been conducted with these haemoparasites. During the present investigation, molecular and microscopic methods were used to screen peripheral blood (n=28) and ectoparasites (n=10 adult ticks n=5 tick nymphs n=1 leech and n>500 tick eggs) collected from wild Tasmanian platypuses (Ornithorhynchus anatinus), for the presence of trypanosomatid-specific DNA and/or trypomastigotes. The genes for the small ribosomal subunit RNA (18S rDNA) and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) were lified and sequenced, prior to conducting phylogenetic analyses. The detection rate of the parasite-specific 18S rDNA in platypus blood was 85.7% (n=24/28), and the leech was also positive at both loci. Microscopically, high parasitaemia and the presence of abundant trypomastigotes, morphologically consistent with Trypanosoma binneyi Mackerras (1959), were observed in the blood films. Phylogenetic analyses at the 18S locus revealed the existence of four trypanosomatid-like genotypes, with variable similarity to two previously-described genotypes of T. binneyi (range of genetic p-distance: 0.0-0.5%). For the gGAPDH locus, for which only one T. binneyi sequence is available in GenBank, three genotypes closely related T. binneyi were identified (range of genetic p-distance: 0.1-0.4%). The leech-derived trypanosome isolate was virtually identical (at the two loci studied) to the other parasites sequenced from infected platypuses however, the molecular or morphological identification of the leech species was not possible. Although further studies are required, the molecular detection of trypanosomes in an aquatic leech removed from a platypus, suggests the possibility that these haematophagous hirudineans may be a vector for T. binneyi (and closely related genotypes).
Publisher: Elsevier BV
Date: 09-2018
DOI: 10.1016/J.IJPARA.2018.05.002
Abstract: Recent molecular and sero-surveillance studies of the tick-borne pathogen Hepatozoon canis have identified new hosts, potential vector species, and have revealed that H. canis is more widespread than previously thought. We report the first diagnosed case of canine hepatozoonosis in Australia from a Maremma Sheepdog in Sarina, Queensland. Hepatozoon canis was detected with blood smear examination and 18S rRNA sequencing. It is unknown when or how the organism was introduced into Australia, which raises questions about border biosecurity policies and the H. canis infection status of its potential vectors and hosts in Australia. Surveillance for this pathogen is required to determine whether H. canis has established in Australia.
Publisher: Elsevier BV
Date: 04-2000
Publisher: Public Library of Science (PLoS)
Date: 13-07-2017
Publisher: Elsevier BV
Date: 06-2016
DOI: 10.1016/J.PARINT.2015.12.005
Abstract: The quokka, Setonix brachyurus, is a vulnerable, small marsupial endemic to Western Australia. Blood s les were collected from quokkas from three different geographical locations Two Peoples Bay, Bald Island and Rottnest Island. The overall prevalence of trypanosomes by nested PCR at the 18S ribosomal RNA gene was 57.3% (63/110) with prevalences of 91.4%, 85.3% and 4.9% respectively for Two Peoples Bay, Bald Island and Rottnest Island. Phylogenetic analysis conducted on 47 18S PCR positives identified two Trypanosoma copemani genotypes, with T. copemani genotype B, the most prevalent genotype infecting quokka populations from the three locations with an overall prevalence of 51.8% (24/47) compared to 34% for T. copemani genotype A (16/47). The overall prevalence of mixed T. copemani genotype A and B infections was 14.9% (7/47). Phylogenetic analysis of 26 quokka isolates at the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) locus, largely supported the 18S analysis but identified a mixed infection in one quokka isolate (Q4112-4117 from Two Peoples Bay). T. copemani genotype B has previously only been isolated from quokkas and the Gilbert's potoroo whereas T. copemani genotype A has a wide host range and may be pathogenic. Further work is required to determine the clinical impact of T. copemani on marsupial populations.
Publisher: Rockefeller University Press
Date: 27-09-1999
Publisher: Elsevier
Date: 2005
Publisher: Elsevier BV
Date: 04-2016
Publisher: Springer Science and Business Media LLC
Date: 10-09-2014
DOI: 10.1007/S00436-014-4118-Z
Abstract: Blood and ectoparasitic ticks were collected from migratory seabirds in New Zealand, including Australasian gannets (n = 13) from two sites and red-billed gulls (n = 9) and white-fronted terns (n = 2) from a third location. Blood smears were screened for parasite presence by microscopy, while DNA from blood s les was subjected to PCR for the presence of tick-transmitted protozoan haemoparasites belonging to the order Piroplasmida. Parasites were identified by comparing small subunit ribosomal RNA (18S rDNA) gene sequences to related sequences on GenBank. Analyses indicated that nine birds were infected with unknown variants of a Babesia poelea-like parasite (recorded as genotypes I and II), while four harboured a piroplasm that was genetically similar to Babesia kiwiensis. There was no parasite stratification by bird species both the gannets and gulls were positive for all three parasites, while the terns were positive for the B. kiwiensis-like and the B. poelea-like (genotype I) parasites. The B. kiwiensis-like parasite found in the birds was also found in two species of ticks: Carios capensis and Ixodes eudyptidis. This represents the first report of Babesia-positive ticks parasitising seabirds in New Zealand. The lack of host specificity and evidence of wide ranging distributions of the three piroplasm genotypes suggests there is a high degree of haemoparasite transmission occurring naturally between New Zealand seabird populations and species.
Publisher: Wiley
Date: 09-05-2019
DOI: 10.1111/TRF.15336
Abstract: A fatal case of autochthonous Babesia microti infection was reported in Australia in 2012. This has implications for Australian public health and, given that babesiosis is transfusion transmissible, has possible implications for Australian blood transfusion recipients. We investigated the seroprevalence of antibodies to B. microti in Australian blood donors and in patients with clinically suspected babesiosis. Plasma s les (n = 7,000) from donors donating in at-risk areas and clinical specimens from patients with clinically suspected babesiosis (n = 29) were tested for B. microti IgG by immunofluorescence assay (IFA). IFA initially reactive s les were tested for B. microti IgG and IgM by immunoblot and B. microti DNA by polymerase chain reaction. Although five donors were initially reactive for B. microti IgG by IFA, none was confirmed for B. microti IgG (zero estimate 95% confidence interval, 0%-0.05%) and all were negative for B. microti DNA. None of the patient s les had B. microti IgG, IgM, or DNA. This study does not provide evidence for widespread exposure to B. microti in Australian blood donors at local theoretical risk, nor does it provide evidence of B. microti infection in Australian patients with clinically suspected babesiosis. Given that confirmed evidence of previous exposure to B. microti was not seen, these data suggest that transmission of this pathogen is currently uncommon in Australia and unlikely to pose a risk to transfusion safety at present.
Publisher: Springer Science and Business Media LLC
Date: 22-02-2019
Publisher: Springer International Publishing
Date: 2019
Publisher: Elsevier BV
Date: 04-2002
Publisher: Palacky University Olomouc
Date: 14-10-2016
DOI: 10.5507/FOT.2015.030
Publisher: Springer Science and Business Media LLC
Date: 04-01-2018
Publisher: Public Library of Science (PLoS)
Date: 02-12-2015
Publisher: Elsevier BV
Date: 05-2000
Publisher: Acumen Publishing Limited
Date: 29-10-2010
Publisher: Elsevier BV
Date: 08-2016
DOI: 10.1016/J.PARINT.2016.03.004
Abstract: The present study describes the first report of Trypanosoma vegrandis in koalas using morphology and sequence analysis of the 18S rRNA gene. The prevalence of T. vegrandis in koalas was 13.6% (6/44). It is likely that the small size of T. vegrandis (<10μm in length), coupled with the difficulties in lifying DNA of this parasite in mixed infections using trypanosome generic primers, are the reason why this organism has not been identified in koalas until now. This study highlights the importance of further research comprising a larger s le size to determine the prevalence of T. vegrandis in koalas as well as its potential impacts upon this marsupial species' health.
Publisher: Elsevier BV
Date: 04-1995
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: Wiley
Date: 2008
DOI: 10.1002/BEM.20399
Abstract: To analyze possible effects of microwaves on gene expression, mice were exposed to global system for mobile communication (GSM) 1800 MHz signal for 1 h at a whole body SAR of 1.1 W/kg. Gene expression was studied in the whole brain, where the average SAR was 0.2 W/kg, by expression microarrays containing over 22,600 probe sets. Comparison of data from sham and exposed animals showed no significant difference in gene expression modulation. However, when less stringent constraints were adopted to analyze microarray results, 75 genes were found to be modulated following exposure. Forty-two probes showed fold changes ranging from 1.5 to 2.8, whereas 33 were down-regulated from 0.67- to 0.29-fold changes, but these differences in gene expression were not confirmed by real-time PCR. Under these specific limited conditions, no consistent indication of gene expression modulation in whole mouse brain was found associated to GSM 1800 MHz exposure.
Publisher: Elsevier BV
Date: 08-2012
DOI: 10.1016/J.EXPPARA.2012.05.009
Abstract: As part of a broader investigation into the potential role of black rats (Rattus rattus) as disease vectors into native small mammal populations of northern Australia, blood and faecal s les from wild black rats were screened by molecular methods, for piroplasms (Babesia and Theileria), trypanosomes and the enteric parasite Cryptosporidium. While piroplasms and trypanosomes were not detected in the blood of these animals, the overall prevalence of Cryptosporidium 18S rDNA in faecal s les was 8.2% (7/85). Co-occurrence of multiple genotypes was observed in 57.1% of the infected in iduals (4/7) cloning and re-sequencing resulted in 14 sequences which broadly grouped with Cryptosporidium sp. rat-genotypes II and III. A novel rat-derived Cryptosporidium sp. genotype at the actin locus was also obtained from five animals. The relatively low infection rate detected, and the epidemiological data on cryptosporidiosis, do not conclusively support a current threat to native Australian mammals from black rats carrying Cryptosporidium. However, this observation is based on s ling limited isolates, in limited regions. Further studies, also including s ling of native mammals, are required on larger s le sizes and from wider geographic areas, to determine the significance of these findings, including the public health importance of Cryptosporidium spp. from rodents.
Publisher: Elsevier BV
Date: 03-2018
DOI: 10.1016/J.TTBDIS.2018.02.001
Abstract: Piroplasms, including the genera Babesia and Theileria, are intra-erythrocytic protozoa that are generally transmitted by ticks and are the aetiological agents for piroplasmosis in animals, as well as humans, worldwide. In Australia, numerous studies have been conducted on piroplasms in domestic animals however, less is known about these protozoa in ticks from native wildlife. The present study characterised piroplasms in Ixodes australiensis (n = 119) and Amblyomma triguttatum (n = 35) ticks collected from kangaroos in Western Australia (WA). Approximately 7.6% (9/119) (95% CI 2.8-12.2) of the I. australiensis ticks were positive for piroplasms using nested-PCR at the 18S rRNA locus, whereas no piroplasm 18S rDNA was detected in the A. triguttatum ticks. All sequences from I. australiensis ticks were identical. Using a 852 bp multiple nucleotide alignment at the 18S rRNA variable region, sequences shared 97.6%, 94.3%, 93.5% and 93.4% pairwise identity with Theileria fuliginosa, Theileria brachyuri, Theileria penicillata, and a Theileria sp. (K1), derived from a burrowing bettong or boodie (Bettongia lesueur), respectively. Phylogenetic analysis revealed that the Theileria sp. from I. australiensis clustered together in the marsupial-associated Theileria group, with T. fuliginosa as closest sister species. Hence, we conclude that this is the first observation of T. fuliginosa-like species in I. australiensis ticks parasitising kangaroos in WA.
Publisher: Public Library of Science (PLoS)
Date: 24-01-2017
Publisher: Elsevier BV
Date: 11-2016
DOI: 10.1016/J.WATRES.2016.09.013
Abstract: Parasites of the genus Cryptosporidium are a major cause of diarrhoea and ill-health in humans and animals and are frequent causes of waterborne outbreaks. Until recently, it was thought that Cryptosporidium was an obligate intracellular parasite that only replicated within a suitable host, and that faecally shed oocysts could survive in the environment but could not multiply. In light of extensive biological and molecular data, including the ability of Cryptosporidium to complete its life cycle in the absence of a host and the production of novel extracellular stages, Cryptosporidium has been formally transferred from the Coccidia, to a new subclass, Cryptogregaria, with gregarine parasites. In this review, we discuss the close relationship between Cryptosporidium and gregarines and discuss the implications for the water industry.
Publisher: Elsevier BV
Date: 09-2017
DOI: 10.1016/J.IJPARA.2017.03.003
Abstract: The extent of within-host genetic ersity of parasites has implications for our understanding of the epidemiology, disease severity and evolution of parasite virulence. As with many other species, our understanding of the within-host ersity of the enteric parasite Cryptosporidium is changing. The present study compared Sanger and Next Generation Sequencing of glycoprotein 60 (gp60) licons from Cryptosporidium hominis (n=11), Cryptosporidium parvum (n=22) and Cryptosporidium cuniculus (n=8) DNA s les from Australia and China. Sanger sequencing identified only one gp60 subtype in each DNA s le: one C. hominis subtype (IbA10G2) (n=11), four C. parvum subtypes belonging to IIa (n=3) and IId (n=19) and one C. cuniculus subtype (VbA23) (n=8). Next Generation Sequencing identified the same subtypes initially identified by Sanger sequencing, but also identified additional gp60 subtypes in C. parvum and C. cuniculus but not in C. hominis, DNA s les. The number of C. parvum and C. cuniculus subtypes identified by Next Generation Sequencing within in idual DNA s les ranged from two to four, and both C. parvum IIa and IId subtype families were identified within the one host in two s les. The finding of the present study has important implications for Cryptosporidium transmission tracking as well as vaccine and drug studies.
Publisher: Elsevier BV
Date: 07-1995
Publisher: Elsevier BV
Date: 09-2002
Publisher: Elsevier BV
Date: 09-2017
Publisher: Informa UK Limited
Date: 07-2006
DOI: 10.1080/03601230600616957
Abstract: The cauliflower mosaic virus 35S promoter (CaMV35s) is extensively used in genetically modified crops for human and animal consumption. Horizontal gene transfer is attracting particular attention, in light of experimental reports, showing the presence of dietary DNA into animal tissues. Health implications may derive from possible activities of the heterologous promoter in mammalian cells after integration in the host genome. To evaluate this hypothesis, in vivo and in vitro experiments were performed using GFP as reporter gene. Recombinant plasmid DNA was fed to Balb/c mice and searched in several tissues by PCR lification. The activity of the plant virus promoter was assessed by RT-PCR and fluorescence microscopy after liposome-mediated transfection of murine gonadic cells. Obtained data did not highlight evidences of dietary DNA transfer in mice. No CaMV35s transcriptional activity was detected in this experimental model. These findings emphasize the need for further studies and standardized methods.
Publisher: BMJ
Date: 06-2020
DOI: 10.1136/BMJOPEN-2019-035930
Abstract: The effect of early and sustained administration of daily probiotic therapy on patients admitted to the intensive care unit (ICU) remains uncertain. The Restoration Of gut microflora in Critical Illness Trial (ROCIT) study is a multicentre, randomised, placebo-controlled, parallel-group, two-sided superiority trial that will enrol 220 patients in five ICUs. Adult patients who are within 48 hours of admission to an ICU and are expected to require intensive care beyond the next calendar day will be randomised in a 1:1 ratio to receive early and sustained Lactobacillus plantarum 299v probiotic therapy in addition to usual care or placebo in addition to usual care. The primary endpoint is days alive and out of hospital to day 60. ROCIT has been approved by the South Metropolitan Health Service Human Research Ethics Committee (ref: RGS00000004) and the St John of God Health Care Human Research Ethics Committee (ref: 1183). The trial results will be submitted for publication in a peer-reviewed journal. Australian and New Zealand Clinical Trials Registry (ANZCTR12617000783325) Pre-results.
Publisher: Elsevier BV
Date: 06-2019
DOI: 10.1016/J.SCITOTENV.2019.03.278
Abstract: Recycled wastewater can carry human-infectious microbial pathogens and therefore wastewater treatment strategies must effectively eliminate pathogens before recycled wastewater is used to supplement drinking and agricultural water supplies. This study characterised the bacterial composition of four wastewater treatment plants (WWTPs) (three waste stabilisation ponds and one oxidation ditch WWTP using activated sludge treatment) in Western Australia. The hypervariable region 4 (V4) of the bacterial 16S rRNA (16S) gene was sequenced using next-generation sequencing (NGS) on the Illumina MiSeq platform. Sequences were pre-processed in USEARCH v10.0 and denoised into zero-radius taxonomic units (ZOTUs) with UNOISE3. Taxonomy was assigned to the ZOTUs using QIIME 2 and the Greengenes database and cross-checked with the NCBI nr/nt database. Bacterial composition of all WWTPs and treatment stages (influent, intermediate and effluent) were dominated by Proteobacteria (29.0-87.4%), particularly Betaproteobacteria (9.0-53.5%) and Gammaproteobacteria (8.6-34.6%). Nitrifying bacteria (Nitrospira spp.) were found only in the intermediate and effluent of the oxidation ditch WWTP, and denitrifying and floc-forming bacteria were detected in all WWTPs, particularly from the families Comamonadaceae and Rhodocyclales. Twelve pathogens were assigned taxonomy by the Greengenes database, but comparison of sequences from genera and families known to contain pathogens to the NCBI nr/nt database showed that only three pathogens (Arcobacter venerupis, Laribacter hongkongensis and Neisseria canis) could be identified in the dataset at the V4 region. Importantly, Enterobacteriaceae genera could not be differentiated. Family level taxa assigned by Greengenes database agreed with NCBI nr/nt in most cases, however, BLAST analyses revealed erroneous taxa in Greengenes database. This study highlights the importance of validating taxonomy of NGS sequences with databases such as NCBI nr/nt, and recommends including the V3 region of 16S in future short licon NGS studies that aim to identify bacterial enteric pathogens, as this will improve taxonomic resolution of most, but not all, Enterobacteriaceae species.
Publisher: Elsevier BV
Date: 2009
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: American Chemical Society (ACS)
Date: 18-01-2007
DOI: 10.1021/JP066294D
Publisher: American Thoracic Society
Date: 07-2022
Publisher: Cushman Foundation for Foraminiferal Research
Date: 1990
DOI: 10.2113/GSJFR.20.1.1
Publisher: Elsevier BV
Date: 02-1994
Publisher: Elsevier BV
Date: 07-1996
Publisher: Elsevier BV
Date: 03-2000
Publisher: American Physical Society (APS)
Date: 07-1993
Publisher: Elsevier BV
Date: 05-2012
DOI: 10.1016/J.EXPPARA.2012.02.021
Abstract: Piroplasms, which include the genera Theileria and Babesia, are blood-borne parasites transmitted mainly by tick vectors. Relatively little is known about their prevalence and clinical impact in Australian marsupials. In the present study the occurrence and molecular phylogeny of these parasites were studied in both wild and captive marsupials from Western Australia (WA) and Queensland (QLD). Blood s les were screened by microscopy and molecular methods, using PCR and DNA sequencing of the 18S ribosomal RNA gene (18S rDNA). Overall, 7.1% of the blood s les (8/113) were positive for piroplasm 18S rDNA. Theileria and Babesia rDNA was detected in 0.9% (1/113) and 6.2% (7/113) of the animals, respectively. The single Theileria positive was identified in one of three boodies (Bettongia lesueur) screened from a wildlife rehabilitation centre in WA, while all seven Babesia positives were detected in WA in wild captured woylies (Bettongia penicillata ogilbyi). Small intraerythrocytic inclusions were observed in blood films made from six of these in iduals. This is the first report of a Babesia sp. in woylies, and Theileria sp. in boodies. Phylogenetic analysis indicated that the woylie-derived Babesia was genetically distinct and most closely related to Babesia occultans, the causative agent of a benign form of cattle babesiosis (genetic similarity 98.4%). The Theileria identified was most closely related to the marsupial-derived species Theileria penicillata from the woylie, Theileria brachyuri from the quokka (Setonix brachyurus), and Theileria sp. from the long-nosed potoroo (Potorous tridactylus).
Publisher: Springer Science and Business Media LLC
Date: 10-02-2021
Publisher: Informa UK Limited
Date: 03-1999
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: Wiley
Date: 23-08-2017
Publisher: Wiley
Date: 09-2016
DOI: 10.1111/PIM.12350
Abstract: Cryptosporidium is a major cause of moderate-to-severe diarrhoea in humans worldwide, second only to rotavirus. Due to the wide host range and environmental persistence of this parasite, cryptosporidiosis can be zoonotic and associated with foodborne and waterborne outbreaks. Currently, 31 species are recognized as valid, and of these, Cryptosporidium hominis and Cryptosporidium parvum are responsible for the majority of infections in humans. The immune status of the host, both innate and adaptive immunity, has a major impact on the severity of the disease and its prognosis. Immunocompetent in iduals typically experience self-limiting diarrhoea and transient gastroenteritis lasting up to 2 weeks and recover without treatment, suggesting an efficient host antiparasite immune response. Immunocompromised in iduals can suffer from intractable diarrhoea, which can be fatal. Effective drug treatments and vaccines are not yet available. As a result of this, the close cooperation and interaction between veterinarians, health physicians, environmental managers and public health operators is essential to properly control this disease. This review focuses on a One Health approach to prophylaxis, including the importance of understanding transmission routes for zoonotic Cryptosporidium species, improved sanitation and better risk management, improved detection, diagnosis and treatment and the prospect of an effective anticryptosporidial vaccine.
Publisher: Elsevier BV
Date: 06-2014
DOI: 10.1016/J.EXPPARA.2014.03.004
Abstract: In 2012, the first autochthonous Australian case of human babesiosis was reported, after microscopic examinations of blood s les revealed intra-erythrocytic parasites in a hospitalized 56year-old man from NSW, who died in 2011 (Senanayake et al., 2012). Independent molecular analyses carried out in Australia and the USA, identified Babesia microti at the 18S ribosomal RNA (18S rRNA), and the beta-tubulin (β-tubulin) gene loci. Here we present the details of a novel PCR-based assay for the β-tubulin gene that was developed, during the original study, to corroborate the results obtained from the analysis of the 18S rDNA. The complete phylogenetic reconstruction, based on the two loci sequenced from the Australian clinical isolate, is also shown here for the first time.
Publisher: Elsevier BV
Date: 1998
DOI: 10.1016/S0006-3495(98)77764-1
Abstract: Menstrual health is an increasingly recognised public health issue, defined as complete physical, mental, and social well-being in relation to the menstrual cycle. The MENISCUS trial aims to assess whether a multi-component intervention addressing physical and emotional aspects of menstrual health improves educational attainment, mental health problems, menstrual management, self-efficacy, and quality of life among girls in secondary school in Uganda. The study is a parallel-arm cluster-randomised controlled trial with 60 schools (clusters) in Wakiso and Kalungu districts, with a mixed-methods process evaluation to assess intervention fidelity and acceptability and economic and policy analyses. The schools will be randomised 1:1 to immediate intervention or to optimised usual care with delayed intervention delivery. The intervention includes creation of a Menstrual Health Action Group at schools and NGO-led training of trainers on puberty education, development of a drama skit, delivery of a menstrual health kit including reusable pads and menstrual cups, access to pain management strategies including analgesics, and basic improvements to school water, sanitation, and hygiene facilities. Baseline data will be collected from secondary 2 students in 2022 (median age ~15.5 years), with endline after 1 year of intervention delivery (~3600 females and a random s le of ~900 males). The primary outcomes assessed in girls are (i) examination performance based on the Mathematics, English, and Biology curriculum taught during the intervention delivery (independently assessed by the Uganda National Examinations Board) and (ii) mental health problems using the Total Difficulties Scale of the Strengths and Difficulties 25-item questionnaire. Secondary outcomes are menstrual knowledge and attitudes in girls and boys and, in girls only, menstrual practices, self-efficacy in managing menstruation, quality of life and happiness, prevalence of urogenital infections, school and class attendance using a self-completed menstrual daily diary, and confidence in maths and science. The trial is innovative in evaluating a multi-component school-based menstrual health intervention addressing both physical and emotional aspects of menstrual health and using a "training of trainers" model designed to be sustainable within schools. If found to be cost-effective and acceptable, the intervention will have the potential for national and regional scale-up. ISRCTN 45461276 . Registered on 16 September 2021.
Publisher: Elsevier BV
Date: 04-2017
DOI: 10.1016/J.VETPAR.2017.04.007
Abstract: A molecular survey was conducted to provide baseline information on the prevalence, genetic ersity and potential clinical impacts of blood-borne and enteric protozoans in native wild mammals from the Northern Territory (NT). A total of 209 blood and 167 faecal s les were collected from four target species the northern brown bandicoot (Isoodon macrourus), common brushtail possum (Trichosurus vulpecula), northern quoll (Dasyurus hallucatus) and brush-tailed rabbit-rat (Conilurus penicillatus). Blood s les were screened by PCR at the 18S rRNA gene for trypanosomes, piroplasms and haemogregarines, with faecal s les tested for Cryptosporidium spp. at the 18S rRNA locus, and for Giardia spp. at the glutamate dehydrogenase (gdh) and 18S rRNA loci. The potential clinical impact was investigated by associating clinical, haematological and biochemical parameters with presence or absence of infection. Overall, 22.5% (95% CI: 17.0-28.8%) of the animals tested were positive for haemoprotozoans. Trypanosomes were found in 26.6% (95% CI: 18.7-35.7%) of the bandicoots and were identified as Trypanosoma vegrandis G6, except for one unique genotype, most similar to T. vegrandis G3 (genetic distance=7%). The prevalence of trypanosomes in possums was 23.7% (95% CI: 11.4-40.2%), and the genotypes identified clustered within the T. noyesi clade. The presence of Babesia sp. and Hepatozoon sp. was confirmed in bandicoots only, both at a prevalence of 9.7% (95% CI: 2.7-9.2%). The total prevalence of intestinal protozoan parasites observed was relatively low (3% 95% CI: 1.0-6.9%). No evidence of clinical disease associated with protozoan parasitic infection was observed, however bandicoots positive for Trypanosoma exhibited a significantly lower packed cell volume (PCV) compared to negative bandicoots (p=0.046). To the authors' knowledge, this is the first research conducted in the NT to characterise protozoan parasites in threatened native mammals using both molecular and morphological tools and to assess the potential clinical impacts of these agents. The absence of clear signs of major morbidity in infected animals seems to exclude a direct association between infections with these agents and possible population decline events in northern Australian native mammals. However until the cause(s) of population decline are ascertained for each in idual mammal species, further studies are required. The outcome of the present investigation may be used to inform wildlife conservation and zoonotic disease programs.
Publisher: Elsevier BV
Date: 02-2018
DOI: 10.1016/J.TTBDIS.2017.12.011
Abstract: Worldwide, Ehrlichia spp. are emerging infectious organisms of domestic animals and people, however, most Ehrlichia spp. naturally infect wildlife reservoirs causing mainly asymptomatic infections. Australian ecosystems have been under-explored for these potentially pathogenic organisms, and recent studies have identified a range of novel Ehrlichia, and their sister genera, Anaplasma and 'Candidatus Neoehrlichia' species, from native Australian ticks. We used bacterial 16S rRNA (16S) next-generation sequencing and genus-specific PCR to profile the bacterial communities in platypus (Ornithorhynchus anatinus) blood s les and platypus ticks (Ixodes ornithorhynchi), and identified a high prevalence of Ehrlichia sequences. We also observed Ehrlichia-like intra-neutrophilic inclusions (morulae) in PCR-positive stained platypus blood films that were consistent in morphology with other Ehrlichia spp. Bayesian phylogenetic analysis of 16S (1343 bp), gltA (1004 bp), and groEL (1074 bp) gene sequences group the platypus Ehrlichia with 'Candidatus Ehrlichia khabarensis' from far-eastern Russia, and demonstrate that the platypus Ehrlichia is clearly distinct from all other Ehrlichia spp. Enough genetic ergence exists to delineate this platypus Ehrlichia as a separate species that we propose to designate 'Candidatus Ehrlichia ornithorhynchi'. There is no evidence that 'Candidatus Ehrlichia ornithorhynchi' causes disease in wild platypuses, however, the organism does seem to be widespread in Australia, being found in both Queensland and Tasmania. 'Candidatus Ehrlichia ornithorhynchi' is the second native Australian Ehrlichia described and adds to the rapidly growing ersity of recently described native Australian tick-borne bacteria.
Publisher: Elsevier BV
Date: 11-2018
DOI: 10.1016/J.SCITOTENV.2018.05.292
Abstract: The cost associated with treatment and disposal of excess sludge produced is one of the greatest operational expenses in wastewater treatment plants. In this study, we quantify and explain greatly reduced excess sludge production in the novel glycogen accumulating organism (GAO) dominated drained biofilm system previously shown to be capable of extremely energy efficient removal of organic carbon (biological oxygen demand or BOD) from wastewater. The average excess sludge production rate was 0.05 g VSS g
Publisher: Informa UK Limited
Date: 12-1995
Publisher: Public Library of Science (PLoS)
Date: 14-12-2016
Publisher: Elsevier BV
Date: 08-2020
Publisher: Public Library of Science (PLoS)
Date: 28-12-2015
Publisher: Elsevier BV
Date: 07-2017
Abstract: Enteric parasites are major contributors to the global diarrhoeal disease load, infecting >67.2 million people. Their prevalence and clinical impact, however, are underestimated due to lack of adequate detection, which is largely still based on microscopy, particularly in developing countries. New commercially available enteric panel assays, which detect parasites (as well as bacteria and/or viruses) using multiplex PCR, offer enhanced sensitivity and specificity as well as the ability to detect mixed infections, and will play an important role in epidemiological surveillance and outbreak investigations. A major limitation of these technologies, however, particularly for developing countries, is the costs involved. Emerging technologies for low-resource, point-of-care (POC) settings have the potential to dramatically improve the cost and accuracy of enteric parasite detection in the future.
Publisher: Elsevier BV
Date: 05-2000
Publisher: Informa UK Limited
Date: 02-05-2007
DOI: 10.1080/09603120701254862
Abstract: A growing number of people attend swimming facilities for recreational activities, rehabilitative treatments, or sport. Filamentous fungi and yeast can be isolated from contaminated air, water and surfaces and may represent a biological risk for employees and users. Here we investigated the occurrence of mycotic species, in a s le of Italian swimming pools (n = 10). Detection and identification of isolated species were achieved by cultural and morphological methods. Results revealed moderate mycotic titres and a high bio ersity. Penicillium spp., Aspergillus spp., Cladosporium spp. and Alternaria sp., were constantly detected in air and surfaces s led by the swimming area, while pathogenic yeast Candida albicans was never detected. Fusarium spp. was the most common taxon isolated from surfaces. For one facility, we typed the genotypic profiles and studied, by genetic typing, the spatial and temporal distribution of isolates. Phylogenetic relationships between species were analysed by alignment of small ribosomal subunit RNA sequences.
Publisher: American Physical Society (APS)
Date: 03-1999
Publisher: Public Library of Science (PLoS)
Date: 22-08-2016
Publisher: Department of Health Science - University of Genoa
Date: 2006
Publisher: Frontiers Media SA
Date: 09-07-2019
Publisher: Elsevier BV
Date: 12-2000
DOI: 10.1016/S0005-2736(00)00330-8
Abstract: We propose a model of calcium channels that can explain most of their observed properties, including the anomalous mole fraction effect and mutation of the glutamate residues. The structure grossly resembles that of the KcsA potassium channel except for the presence of an extracellular vestibule and a shorter selectivity filter containing four glutamate residues. Using this model in electrostatic calculations and Brownian dynamics simulations, we study mechanisms of ion permeation and selectivity in the channel. Potential energy profiles calculated for multiple ions in the channel provide explanations of ion permeation, the block of Na(+) currents by Ca(2+) ions, and many other observed properties. Brownian dynamics simulations provide quantitative predictions for the channel currents which reproduce available experimental data.
Publisher: Elsevier BV
Date: 2001
Publisher: Wiley
Date: 03-2012
DOI: 10.5694/MJA11.11378
Publisher: Elsevier BV
Date: 03-2017
DOI: 10.1016/J.BIORTECH.2016.11.102
Abstract: Glycogen accumulating organisms (GAO) are known to allow anaerobic uptake of biological oxygen demand (BOD) in activated sludge wastewater treatment systems. In this study, we report a rapid transition of suspended activated sludge biomass to a GAO dominated biofilm by selective enrichment using sequences of anaerobic loading followed by aerobic exposure of the biofilm to air. The study showed that within eight weeks, a fully operational, GAO dominated biofilm had developed, enabling complete anaerobic BOD uptake at a rate of 256mg/L/h. The oxygen uptake by the biofilm directly from the atmosphere had been calculated to provide significant energy savings. This study suggests that wastewater treatment plant operators can convert activated sludge systems readily into a "passive aeration" biofilm that avoids costly oxygen transfer to bulk wastewater solution. The described energy efficient BOD removal system provides an opportunity to be coupled with novel nitrogen removal processes such as anammox.
Publisher: Informa UK Limited
Date: 03-2000
Publisher: Elsevier BV
Date: 12-2015
DOI: 10.1016/J.PROTIS.2015.10.001
Abstract: Piroplasms, tick-transmitted Apicomplexa of the genera Theileria, Babesia and Cytauxzoon, are blood-borne parasites of clinical and veterinary importance. The order Piroplasmida shows a puzzling systematics characterized by multiple clades, soft polytomies and paraphyletic olyphyletic genera. In the present study, screening of platypuses (Ornithorhynchus anatinus), was performed to infer the parasite molecular phylogeny. DNA was extracted from blood, ectoparasites and tick eggs and the 18S rRNA- hsp70-genes were used for the phylogenetic reconstructions. Microscopic analyses detected pleomorphic intra-erythrocytic organisms and tetrads consistent with previous descriptions of Theileria ornithorhynchi Mackerras, 1959, but observation of possible schizonts could not be confirmed. DNA sequences obtained from blood and ticks allowed resolving the systematics of the first piroplasm infecting a monotreme host. Molecularly, T. ornithorhynchi formed a novel monophyletic group, basal to most known piroplasms' clades. The ancestral position of this clade, isolated from an ancient lineage of mammalian host appears particularly fascinating. The present paper discusses the inadequacies of the current molecular systematics for the Piroplasmida and the consequences of incomplete s ling, morphology-based classification and ambiguous microscopic identifications. Likely when the current s ling bias is rectified and more sequence data is made available, the phylogenetic position of T. ornithorhynchi will be further contextualized without ambiguity.
Publisher: Ovid Technologies (Wolters Kluwer Health)
Date: 05-2007
Publisher: Springer Science and Business Media LLC
Date: 25-06-2015
Publisher: Cambridge University Press (CUP)
Date: 14-08-2017
DOI: 10.1017/S0031182017001214
Abstract: Currently, the systematics, biology and epidemiology of piscine Cryptosporidium species are poorly understood. Here, we compared Sanger ‒ and next-generation ‒ sequencing (NGS), of piscine Cryptosporidium , at the 18S rRNA and actin genes. The hosts comprised 11 ornamental fish species, spanning four orders and eight families. The objectives were: to (i) confirm the rich genetic ersity of the parasite and the high frequency of mixed infections and (ii) explore the potential of NGS in the presence of complex genetic mixtures. By Sanger sequencing, four main genotypes were obtained at the actin locus, while for the 18S locus, seven genotypes were identified. At both loci, NGS revealed frequent mixed infections, consisting of one highly dominant variant plus substantially rarer genotypes. Both sequencing methods detected novel Cryptosporidium genotypes at both loci, including a novel and highly abundant actin genotype that was identified by both Sanger sequencing and NGS. Importantly, this genotype accounted for 68·9% of all NGS reads from all s les (249 585/362 372). The present study confirms that aquarium fish can harbour a large and unexplored Cryptosporidium genetic ersity. Although commonly used in molecular parasitology studies, nested PCR prevents quantitative comparisons and thwarts the advantages of NGS, when this latter approach is used to investigate multiple infections.
Publisher: Elsevier BV
Date: 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
Date: 2004
Publisher: Elsevier BV
Date: 07-2019
DOI: 10.1016/J.WATRES.2019.04.041
Abstract: While some microbial eukaryotes can improve effluent quality in wastewater treatment plants (WWTPs), eukaryotic waterborne pathogens are a threat to public health. This study aimed to identify Eukarya, particularly faecal pathogens including Cryptosporidium, in different treatment stages (influent, intermediate and effluent) from four WWTPs in Western Australia (WA). Three WWTPs that utilise stabilisation ponds and one WWTP that uses activated sludge (oxidation ditch) treatment technologies were s led. Eukaryotic 18S rRNA (18S) was targeted in the wastewater s les (n = 26) for next-generation sequencing (NGS), and a mammalian-blocking primer was used to reduce the lification of mammalian DNA. Overall, bioinformatics analyses revealed 49 eukaryotic phyla in WWTP s les, and three of these phyla contained human intestinal parasites, which were primarily detected in the influent. These human intestinal parasites either had a low percent sequence composition or were not detected in the intermediate and effluent stages and included the amoebozoans Endolimax sp., Entamoeba sp. and Iodamoeba sp., the human pinworm Enterobius vermicularis (Nematoda), and Blastocystis sp. subtypes (Sarcomastigophora). Six Blastocystis subtypes and four Entamoeba species were identified by eukaryotic 18S NGS, however, Cryptosporidium sp. and Giardia sp. were not detected. Real-time polymerase chain reaction (PCR) also failed to detect Giardia, but Cryptosporidium-specific NGS detected Cryptosporidium in all WWTPs, and a total of nine species were identified, including five zoonotic pathogens. Although eukaryotic 18S NGS was able to identify some faecal pathogens, this study has demonstrated that more specific NGS approaches for pathogen detection are more sensitive and should be applied to future wastewater pathogen assessments.
Publisher: Elsevier BV
Date: 04-1995
Publisher: Springer Science and Business Media LLC
Date: 10-04-2015
Publisher: Elsevier BV
Date: 08-2014
DOI: 10.1016/J.WATRES.2014.04.023
Abstract: Reliable identification of cyanobacterial isolates has significant socio-economic implications as many bloom-forming species affect the aesthetics and safety of drinking water, through the production of taste and odour compounds or toxic metabolites. The limitations of morphological identification have promoted the application of molecular tools, and encouraged the adoption of combined (polyphasic) approaches that include both microscopy- and DNA-based analyses. In this context, the rapid expansion of available sequence data is expected to allow increasingly reliable identification of cyanobacteria, and ultimately resolve current discrepancies between the two approaches. In the present study morphological and molecular characterisations of cyanobacterial isolates (n = 39), collected from various freshwater sites in Australia, were compared. Sequences were obtained for the small ribosomal subunit RNA gene (16S rDNA) (n = 36), the DNA-dependent RNA polymerase gene (rpoC1) (n = 22), and the phycocyanin operon, with its intergenic spacer region (cpcBA-IGS) (n = 19). Phylogenetic analyses identified three cyanobacterial orders: the Chroococcales (n = 8), Oscillatoriales (n = 6), and Nostocales (n = 25). Interestingly, multiple novel genotypes were identified, with 22% of the strains (17/77) having <95% similarity to available sequences in GenBank. Morphological and molecular data were in agreement at the species level for only 26% of the isolates obtained (10/39), while agreement at the genus level was obtained for 31% (12/39). Confident identification of the remaining 44% of the strains (17/39) beyond the order level was not possible. The present study demonstrates that, despite the taxonomic revisions, and advances in molecular-, and bioinformatics-tools, the lack of reliable morphological features, culture-induced pleomorphism, and proportion of misidentified or poorly described sequences in GenBank, still represent significant factors, impeding the confident identification of cyanobacteria species.
Publisher: Elsevier BV
Date: 12-2011
DOI: 10.1016/J.VETPAR.2011.07.009
Abstract: In the present study, the occurrence and molecular phylogeny of trypanosome parasites were studied in both wild and captive marsupials from Western Australia and Queensland. Blood s les were screened by PCR at the 18S rDNA locus, and the glycosomal glyceraldehyde phosphate dehydrogenase gene. Overall, 5.3% of the blood s les were positive at the 18S rDNA locus. All positives belonged to wild-captured Western Australian in iduals, where trypanosome-specific DNA was detected in 9.8% of the screened s les from wild marsupials, in common brushtail possums, and woylies. The detection rate of trypanosome DNA in these two host species was 12.5% and 20%, respectively. Phylogenetic analyses based on two loci, indicated that the possum-derived trypanosome isolates were genetically distinct, and most closely related to the Australian marsupial trypanosomes H25 from a kangaroo, and BRA2 from a bush rat. This is the first study to genetically characterise trypanosome isolates from possums. The analysis of the woylie-derived isolates demonstrated that this marsupial host can harbour multiple genotypes within the same geographical location and furthermore multiple genotypes within the same host, indicative of mixed infections. All the woylie-derived genotypes grouped with trypanosomes found in Australian marsupials, suggesting that they are more likely to belong to an endemic or Australasian trypanosome species. This is the first study to genetically characterise trypanosome isolates from possums (Trichosurus vulpecula). Although the clinical significance of these infections is currently unknown, the identification of these novel sequences may support future investigations on transmission, threats to endangered wildlife, and evolutionary history of the genus Trypanosoma.
No related grants have been discovered for Andrea Paparini.