Publications by authors named "Matthew J Nolan"

48 Publications

Two new species of Prosorhynchoides (Digenea: Bucephalidae) from Tylosurus crocodilus (Belonidae) from the great barrier reef and French Polynesia.

Parasitol Int 2020 Apr 28;75:102005. Epub 2019 Oct 28.

School of Science, Centre for Environmental Sustainability and Remediation, RMIT University, PO Box 71, Bundoora, Victoria 3083, Australia. Electronic address:

We surveyed 14 individuals of Tylosurus crocodilus Péron & Lesueur 1821 (Belonidae) collected from the waters around Lizard Island and Heron Island, Great Barrier Reef, Queensland, Australia, and the waters around Moorea, French Polynesia. We describe two new species of bucephaline trematodes from them, Prosorhynchoides galaktionovi n. sp. and P. kohnae n. sp. They are morphologically distinct from existing Prosorhynchoides spp., with molecular data from 28S and ITS-2 ribosomal DNA, as well as cox1 mitochondrial DNA, further supporting our morphological findings. Neither species has been observed in other belonid fishes. The new species fall into the clade of species of Prosorhynchoides from belonids previously identified in Australian waters. These findings strengthen the observation that groups of bucephaline species have radiated, at least in part, in tight association with host taxa. There are now five species of Prosorhynchoides known from two belonid species in Australian waters. We, therefore, predict further richness in the nine other belonid species present.
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http://dx.doi.org/10.1016/j.parint.2019.102005DOI Listing
April 2020

A C-Terminal Fragment of Chlorotoxin Retains Bioactivity and Inhibits Cell Migration.

Front Pharmacol 2019 20;10:250. Epub 2019 Mar 20.

Centre for Molecular Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, QLD, Australia.

Chlorotoxin was originally isolated from the venom of the Israeli scorpion , and has potential as a tumor imaging agent based on its selective binding to tumor cells. Several targets have been suggested for chlorotoxin including voltage-gated chloride channels, and it has been shown to have anti-angiogenic activity and inhibit cell migration. The structure of chlorotoxin is stabilized by four disulfide bonds and contains β-sheet and helical structure. Interestingly, the reduced form has previously been shown to inhibit cell migration to the same extent as the wild type, but structural analysis indicates that the reduced form of the peptide does not maintain the native secondary structure and appears unstructured in solution. This lack of structure suggests that a short stretch of amino acids might be responsible for the bioactivity. To explore this hypothesis, we have synthesized fragments of chlorotoxin without disulfide bonds. As expected for such small peptides, NMR analysis indicated that the peptides were unstructured in solution. However, the peptide corresponding to the eight C-terminal residues inhibited cell migration, in contrast to the other fragments. Our results suggest that the C-terminal region plays a critical role in the bioactivity of chlorotoxin.
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http://dx.doi.org/10.3389/fphar.2019.00250DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6435586PMC
March 2019

Dissecting the Genomic Architecture of Resistance to Parasitism in the Chicken.

Front Genet 2018 26;9:528. Epub 2018 Nov 26.

The Roslin Institute and Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Edinburgh, United Kingdom.

Coccidiosis in poultry, caused by protozoan parasites of the genus , is an intestinal disease with substantial economic impact. With the use of anticoccidial drugs under public and political pressure, and the comparatively higher cost of live-attenuated vaccines, an attractive complementary strategy for control is to breed chickens with increased resistance to parasitism. Prior infection with leads to complete immunity against challenge with homologous strains, but only partial resistance to challenge with antigenically diverse heterologous strains. We investigate the genetic architecture of avian resistance to primary infection and heterologous strain secondary challenge using White Leghorn populations of derived inbred lines, C.B12 and 15I, known to differ in susceptibility to the parasite. An intercross population was infected with Houghton (H) strain, followed 3 weeks later by Weybridge (W) strain challenge, while a backcross population received a single W infection. The phenotypes measured were parasite replication (counting fecal oocyst output or qPCR for parasite numbers in intestinal tissue), intestinal lesion score (gross pathology, scale 0-4), and for the backcross only, serum interleukin-10 (IL-10) levels. Birds were genotyped using a high density genome-wide DNA array (600K, Affymetrix). Genome-wide association study located associations on chromosomes 1, 2, 3, and 5 following primary infection in the backcross population, and a suggestive association on chromosome 1 following heterologous W challenge in the intercross population. This mapped several megabases away from the quantitative trait locus (QTL) linked to the backcross primary W strain infection, suggesting different underlying mechanisms for the primary- and heterologous secondary- responses. Underlying pathways for those genes located in the respective QTL for resistance to primary infection and protection against heterologous challenge were related mainly to immune response, with IL-10 signaling in the backcross primary infection being the most significant. Additionally, the identified markers associated with IL-10 levels exhibited significant additive genetic variance. We suggest this is a phenotype of interest to the outcome of challenge, being scalable in live birds and negating the requirement for single-bird cages, fecal oocyst counts, or slaughter for sampling (qPCR).
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http://dx.doi.org/10.3389/fgene.2018.00528DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6275401PMC
November 2018

Phenotypic and genetic variation in the response of chickens to Eimeria tenella induced coccidiosis.

Genet Sel Evol 2018 Nov 21;50(1):63. Epub 2018 Nov 21.

The Roslin Institute, University of Edinburgh, Easter Bush, Midlothian, UK.

Background: Coccidiosis is a major contributor to losses in poultry production. With emerging constraints on the use of in-feed prophylactic anticoccidial drugs and the relatively high costs of effective vaccines, there are commercial incentives to breed chickens with greater resistance to this important production disease. To identify phenotypic biomarkers that are associated with the production impacts of coccidiosis, and to assess their covariance and heritability, 942 Cobb500 commercial broilers were subjected to a defined challenge with Eimeria tenella (Houghton). Three traits were measured: weight gain (WG) during the period of infection, caecal lesion score (CLS) post mortem, and the level of a serum biomarker of intestinal inflammation, i.e. circulating interleukin 10 (IL-10), measured at the height of the infection.

Results: Phenotypic analysis of the challenged chicken cohort revealed a significant positive correlation between CLS and IL-10, with significant negative correlations of both these traits with WG. Eigenanalysis of phenotypic covariances between measured traits revealed three distinct eigenvectors. Trait weightings of the first eigenvector, (EV1, eigenvalue = 59%), were biologically interpreted as representing a response of birds that were susceptible to infection, with low WG, high CLS and high IL-10. Similarly, the second eigenvector represented infection resilience/resistance (EV2, 22%; high WG, low CLS and high IL-10), and the third eigenvector tolerance (EV3, 19%; high WG, high CLS and low IL-10), respectively. Genome-wide association studies (GWAS) identified two SNPs that were associated with WG at the suggestive level.

Conclusions: Eigenanalysis separated the phenotypic impact of a defined challenge with E. tenella on WG, caecal inflammation/pathology, and production of IL-10 into three major eigenvectors, indicating that the susceptibility-resistance axis is not a single continuous quantitative trait. The SNPs identified by the GWAS for body weight were located in close proximity to two genes that are involved in innate immunity (FAM96B and RRAD).
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http://dx.doi.org/10.1186/s12711-018-0433-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6249784PMC
November 2018

Heterobucephalopsine and prosorhynchine trematodes (Digenea: Bucephalidae) from teleost fishes of Moreton Bay, Queensland, Australia, with the description of two new species.

Syst Parasitol 2018 11 3;95(8-9):783-806. Epub 2018 Oct 3.

School of Biological Sciences, The University of Queensland, Brisbane, QLD, 4072, Australia.

Eight species of the trematode family Bucephalidae Poche, 1907 are reported from teleost fishes in Moreton Bay, Queensland, Australia. Heterobucephalopsis yongi n. sp. is described from Gymnothorax eurostus (Muraenidae); the new form is distinguished from its congeners in the possession of a tiny cirrus-sac relative to body length, the length of the caecum, the position of the mouth and pharynx, and the position of the testes and ovary. Two known species of Dollfustrema Eckmann, 1934, D. durum Nolan, Curran, Miller, Cutmore, Cantacessi & Cribb, 2015 and D. gibsoni Nolan & Cribb, 2010, are reported from Gymnothorax pseudothyrsoideus (Bleeker) (Muraenidae); although both species were described from Australian waters, this represents the first reports from Moreton Bay and G. pseudothyrsoideus. Four species of Prosorhynchus Odhner, 1905 are reported, including one new, P. brayi n. sp., which is described from Epinephelus coioides (Hamilton) (Serranidae); P. brayi n. sp. is distinguished from its congeners in the possession of vitelline follicles in a confluent arc distinctly posterior to a conical rhynchus, uterine coils that do not extend anterior to the vitelline arc, contiguous testes, a cirrus-sac that reaches anteriorly to at least the level of the posterior testis and a short excretory vesicle. Three known species of Prosorhynchus are reported from Australia, for the first time: P. luzonicus Velasquez, 1959 and P. maternus Bray & Justine, 2006 from E. coioides and Prosorhynchus platycephali (Yamaguti, 1934) Srivastava, 1938 from Ambiserrula jugosa (McCulloch) and Inegocia japonica (Cuvier) (Platycephalidae). Skrjabiniella Issaitschikow, 1928 is re-recognised for new specimens of Skrjabiniella uniporus (Ozaki, 1924) n. comb. collected from Conger cinereus Rüppell (Congridae); three additional species of Prosorhynchus are considered members of this genus, two of which are synonymised with S. uniporus.
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http://dx.doi.org/10.1007/s11230-018-9820-2DOI Listing
November 2018

Does selection for growth rate in broilers affect their resistance and tolerance to Eimeria maxima?

Vet Parasitol 2018 Jul 18;258:88-98. Epub 2018 Jun 18.

School of Natural and Environmental Sciences, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK.

Chickens exhibit varied responses to infection with Eimeria parasites. We hypothesise that broilers selected for increased growth rate will show lower resistance and tolerance to a coccidian challenge. 288 chickens of fast (F) or slow (S) growing lines were inoculated with 0 (control), 2500 (low-dose), or 7000 (high-dose) sporulated E. maxima oocysts at 13 days of age in two consecutive rounds. Gain and Intake were measured daily and their values relative to BW at the point of infection were calculated over the pre-patent (days 1-4 post-infection), acute (d5-8 pi), and recovery (d9-12 pi) phases of infection to assess the impact of infection. Levels of plasma carotenoids, vitamins E and A, long bone mineralisation, caecal microbiota diversity indices, and histological measurements were assessed at the acute (d6 pi) and recovery stage (d13 pi). In addition, we measured the levels of nitric oxide metabolites and the number of parasite genome copies in the jejunumat d6pi. In absolute terms F birds grew 1.42 times faster than S birds when not infected. Infection significantly reduced relative daily gain and intake (P < 0.001), with the effects being most pronounced during the acute phase (P < 0.001). Levels of all metabolites were significantly decreased, apart from NO which increased (P < 0.001) in response to infection on d6pi, and were accompanied by changes in histomorphometric features and the presence of E. maxima genome copies in infected birds, which persisted to d13pi. Furthermore, infection reduced tibia and femur mineralisation, which also persisted to d13pi. Reductions in measured variables were mostly independent of dose size, as was the level of parasite replication. The impact of infection was similar for S and F-line birds for all measured parameters, and there were no significant interactions between line x dose size on any of these parameters. In conclusion, our results suggest that line differences in productive performance do not influence host responses to coccidiosis when offered nutrient adequate diets.
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http://dx.doi.org/10.1016/j.vetpar.2018.06.014DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6052249PMC
July 2018

The Aromatic Head Group of Spider Toxin Polyamines Influences Toxicity to Cancer Cells.

Toxins (Basel) 2017 10 27;9(11). Epub 2017 Oct 27.

Centre for Biodiscovery and Molecular Development of Therapeutics, AITHM, James Cook University, Cairns, QLD 4878, Australia.

Spider venoms constitute incredibly diverse libraries of compounds, many of which are involved in prey capture and defence. Polyamines are often prevalent in the venom and target ionotropic glutamate receptors. Here we show that a novel spider polyamine, PA, containing a hydroxyphenyl-based structure is present in the venom of several species of tarantula, and has selective toxicity against MCF-7 breast cancer cells. By contrast, a polyamine from an Australian funnel-web spider venom, which contains an identical polyamine tail to PA but an indole-based head-group, is only cytotoxic at high concentrations. Our results suggest that the ring structure plays a role in the cytotoxicity and that modification to the polyamine head group might lead to more potent and selective compounds with potential as novel cancer treatments.
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http://dx.doi.org/10.3390/toxins9110346DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5705961PMC
October 2017

Recombinant anticoccidial vaccines - a cup half full?

Infect Genet Evol 2017 11 7;55:358-365. Epub 2017 Oct 7.

Department of Pathobiology and Population Sciences, Royal Veterinary College, University of London, Hawkshead Lane, Hertfordshire AL9 7TA, UK.

Eimeria species parasites can cause the disease coccidiosis, most notably in chickens. The occurrence of coccidiosis is currently controlled through a combination of good husbandry, chemoprophylaxis and/or live parasite vaccination; however, scalable, cost-effective subunit or recombinant vaccines are required. Many antigens have been proposed for use in novel anticoccidial vaccines, supported by the capacity to reduce disease severity or parasite replication, increase body weight gain in the face of challenge or improve feed conversion under experimental conditions, but none has reached commercial development. Nonetheless, the protection against challenge induced by some antigens has been within the lower range described for the ionophores against susceptible isolates or current live vaccines prior to oocyst recycling. With such levels of efficacy it may be that combinations of anticoccidial antigens already described are sufficient for development as novel multi-valent vaccines, pending identification of optimal delivery systems. Selection of the best antigens to be included in such vaccines can be informed by knowledge defining the natural occurrence of specific antigenic diversity, with relevance to the risk of immediate vaccine breakthrough, and the rate at which parasite genomes can evolve new diversity. For Eimeria, such data are now becoming available for antigens such as apical membrane antigen 1 (AMA1) and immune mapped protein 1 (IMP1) and more are anticipated as high-capacity, high-throughput sequencing technologies become increasingly accessible.
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http://dx.doi.org/10.1016/j.meegid.2017.10.009DOI Listing
November 2017

Effects of Eimeria tenella infection on chicken caecal microbiome diversity, exploring variation associated with severity of pathology.

PLoS One 2017 21;12(9):e0184890. Epub 2017 Sep 21.

Department of Pathobiology and Population Sciences, Royal Veterinary College, University of London, Hatfield, United Kingdom.

Eimeria species cause the intestinal disease coccidiosis, most notably in poultry. While the direct impact of coccidiosis on animal health and welfare is clear, its influence on the enteric microbiota and by-stander effects on chicken health and production remains largely unknown, with the possible exception of Clostridium perfringens (necrotic enteritis). This study evaluated the composition and structure of the caecal microbiome in the presence or absence of a defined Eimeria tenella challenge infection in Cobb500 broiler chickens using 16S rRNA amplicon sequencing. The severity of clinical coccidiosis in individual chickens was quantified by caecal lesion scoring and microbial changes associated with different lesion scores identified. Following E. tenella infection the diversity of taxa within the caecal microbiome remained largely stable. However, infection induced significant changes in the abundance of some microbial taxa. The greatest changes were detected in birds displaying severe caecal pathology; taxa belonging to the order Enterobacteriaceae were increased, while taxa from Bacillales and Lactobacillales were decreased with the changes correlated with lesion severity. Significantly different profiles were also detected in infected birds which remained asymptomatic (lesion score 0), with taxa belonging to the genera Bacteroides decreased and Lactobacillus increased. Many differential taxa from the order Clostridiales were identified, with some increasing and others decreasing in abundance in Eimeria-infected animals. The results support the view that caecal microbiome dysbiosis associated with Eimeria infection contributes to disease pathology, and could be a target for intervention to mitigate the impact of coccidiosis on poultry productivity and welfare. This work highlights that E. tenella infection has a significant impact on the abundance of some caecal bacteria with notable differences detected between lesion score categories emphasising the importance of accounting for differences in caecal lesions when investigating the relationship between E. tenella and the poultry intestinal microbiome.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0184890PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5608234PMC
October 2017

Infections by human gastrointestinal helminths are associated with changes in faecal microbiota diversity and composition.

PLoS One 2017 11;12(9):e0184719. Epub 2017 Sep 11.

Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom.

Investigations of the impact that patent infections by soil-transmitted gastrointestinal nematode parasites exert on the composition of the host gut commensal flora are attracting growing interest by the scientific community. However, information collected to date varies across experiments, and further studies are needed to identify consistent relationships between parasites and commensal microbial species. Here, we explore the qualitative and quantitative differences between the microbial community profiles of cohorts of human volunteers from Sri Lanka with patent infection by one or more parasitic nematode species (H+), as well as that of uninfected subjects (H-) and of volunteers who had been subjected to regular prophylactic anthelmintic treatment (Ht). High-throughput sequencing of the bacterial 16S rRNA gene, followed by bioinformatics and biostatistical analyses of sequence data revealed no significant differences in alpha diversity (Shannon) and richness between groups (P = 0.65, P = 0.13 respectively); however, beta diversity was significantly increased in H+ and Ht when individually compared to H-volunteers (P = 0.04). Among others, bacteria of the families Verrucomicrobiaceae and Enterobacteriaceae showed a trend towards increased abundance in H+, whereas the Leuconostocaceae and Bacteroidaceae showed a relative increase in H- and Ht respectively. Our findings add valuable knowledge to the vast, and yet little explored, research field of parasite-microbiota interactions and will provide a basis for the elucidation of the role such interactions play in pathogenic and immune-modulatory properties of parasitic nematodes in both human and animal hosts.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0184719PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5593201PMC
October 2017

Molecular characterisation of protist parasites in human-habituated mountain gorillas (Gorilla beringei beringei), humans and livestock, from Bwindi impenetrable National Park, Uganda.

Parasit Vectors 2017 Jul 18;10(1):340. Epub 2017 Jul 18.

Department of Pathobiology and Population Sciences, Royal Veterinary College, University of London, Hatfield, UK.

Background: Over 60 % of human emerging infectious diseases are zoonotic, and there is growing evidence of the zooanthroponotic transmission of diseases from humans to livestock and wildlife species, with major implications for public health, economics, and conservation. Zooanthroponoses are of relevance to critically endangered species; amongst these is the mountain gorilla (Gorilla beringei beringei) of Uganda. Here, we assess the occurrence of Cryptosporidium, Cyclospora, Giardia, and Entamoeba infecting mountain gorillas in the Bwindi Impenetrable National Park (BINP), Uganda, using molecular methods. We also assess the occurrence of these parasites in humans and livestock species living in overlapping/adjacent geographical regions.

Results: Diagnostic PCR detected Cryptosporidium parvum in one sample from a mountain gorilla (IIdA23G2) and one from a goat (based on SSU). Cryptosporidium was not detected in humans or cattle. Cyclospora was not detected in any of the samples analysed. Giardia was identified in three human and two cattle samples, which were linked to assemblage A, B and E of G. duodenalis. Sequences defined as belonging to the genus Entamoeba were identified in all host groups. Of the 86 sequence types characterised, one, seven and two have been recorded previously to represent genotypes of Cryptosporidium, Giardia, and Entamoeba, respectively, from humans, other mammals, and water sources globally.

Conclusions: This study provides a snapshot of the occurrence and genetic make-up of selected protists in mammals in and around BINP. The genetic analyses indicated that 54.6% of the 203 samples analysed contained parasites that matched species, genotypes, or genetic assemblages found globally. Seventy-six new sequence records were identified here for the first time. As nothing is known about the zoonotic/zooanthroponotic potential of the corresponding parasites, future work should focus on wider epidemiological investigations together with continued surveillance of all parasites in humans, other mammals, the environment, and water in this highly impoverished area.
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http://dx.doi.org/10.1186/s13071-017-2283-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5516388PMC
July 2017

Helminth infections and gut microbiota - a feline perspective.

Parasit Vectors 2016 12 3;9(1):625. Epub 2016 Dec 3.

Department of Veterinary Medicine, University of Cambridge, Cambridge, UK.

Background: Investigations of the relationships between the gut microbiota and gastrointestinal parasitic nematodes are attracting growing interest by the scientific community, driven by the need to better understand the contribution of parasite-associated changes in the composition of the gut flora to both host malnutrition and immune modulation. These studies have however been carried out mainly in humans and experimental animals, while knowledge of the make-up of the gut commensal flora in presence or absence of infection by parasitic nematodes in domestic animals is limited. In this study, we investigate the qualitative and quantitative impact that infections by a widespread parasite of cats (i.e. Toxocara cati) exert on the gut microbiota of feline hosts.

Methods: The faecal microbiota of cats with patent infection by T. cati (= Tc+), as well as that of negative controls (= Tc-) was examined via high-throughput sequencing of the V3-V4 hypervariable region of the bacterial 16S rRNA gene, followed by bioinformatics and biostatistical analyses of sequence data.

Results: A total of 2,325,366 useable high-quality sequences were generated from the faecal samples analysed in this study and subjected to further bioinformatics analyses, which led to the identification of 128 OTUs and nine bacterial phyla, respectively. The phylum Firmicutes was predominant in all samples analysed (mean of 53.0%), followed by the phyla Proteobacteria (13.8%), Actinobacteria (13.7%) and Bacteroidetes (10.1%). Among others, bacteria of the order Lactobacillales, the family Enterococcaceae and genera Enterococcus and Dorea showed a trend towards increased abundance in Tc+  compared with Tc- samples, while no significant differences in OTU richness and diversity were recorded between Tc+ and Tc- samples (P = 0.485 and P = 0.581, respectively). However, Canonical Correlation and Redundancy Analyses were able to separate samples by infection status (P = 0.030 and P = 0.015, respectively), which suggests a correlation between the latter and the composition of the feline faecal microbiota.

Conclusions: In spite of the relatively small number of samples analysed, subtle differences in the composition of the gut microbiota of Tc+ vs Tc- cats could be identified, some of which in accordance with current data from humans and laboratory animal hosts. Nevertheless, the findings from this study contribute valuable knowledge to the yet little explored area of parasite-microbiota interactions in domestic animals.
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http://dx.doi.org/10.1186/s13071-016-1908-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5135779PMC
December 2016

The Anisakis Transcriptome Provides a Resource for Fundamental and Applied Studies on Allergy-Causing Parasites.

PLoS Negl Trop Dis 2016 07 29;10(7):e0004845. Epub 2016 Jul 29.

Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom.

Background: Food-borne nematodes of the genus Anisakis are responsible for a wide range of illnesses (= anisakiasis), from self-limiting gastrointestinal forms to severe systemic allergic reactions, which are often misdiagnosed and under-reported. In order to enhance and refine current diagnostic tools for anisakiasis, knowledge of the whole spectrum of parasite molecules transcribed and expressed by this parasite, including those acting as potential allergens, is necessary.

Methodology/principal Findings: In this study, we employ high-throughput (Illumina) sequencing and bioinformatics to characterise the transcriptomes of two Anisakis species, A. simplex and A. pegreffii, and utilize this resource to compile lists of potential allergens from these parasites. A total of ~65,000,000 reads were generated from cDNA libraries for each species, and assembled into ~34,000 transcripts (= Unigenes); ~18,000 peptides were predicted from each cDNA library and classified based on homology searches, protein motifs and gene ontology and biological pathway mapping. Using comparative analyses with sequence data available in public databases, 36 (A. simplex) and 29 (A. pegreffii) putative allergens were identified, including sequences encoding 'novel' Anisakis allergenic proteins (i.e. cyclophilins and ABA-1 domain containing proteins).

Conclusions/significance: This study represents a first step towards providing the research community with a curated dataset to use as a molecular resource for future investigations of the biology of Anisakis, including molecules putatively acting as allergens, using functional genomics, proteomics and immunological tools. Ultimately, an improved knowledge of the biological functions of these molecules in the parasite, as well as of their immunogenic properties, will assist the development of comprehensive, reliable and robust diagnostic tools.
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http://dx.doi.org/10.1371/journal.pntd.0004845DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4966942PMC
July 2016

Cryptic Eimeria genotypes are common across the southern but not northern hemisphere.

Int J Parasitol 2016 08 29;46(9):537-44. Epub 2016 Jun 29.

Department of Pathology and Pathogen Biology, Royal Veterinary College, North Mymms, Hertfordshire, UK. Electronic address:

The phylum Apicomplexa includes parasites of medical, zoonotic and veterinary significance. Understanding the global distribution and genetic diversity of these protozoa is of fundamental importance for efficient, robust and long-lasting methods of control. Eimeria spp. cause intestinal coccidiosis in all major livestock animals and are the most important parasites of domestic chickens in terms of both economic impact and animal welfare. Despite having significant negative impacts on the efficiency of food production, many fundamental questions relating to the global distribution and genetic variation of Eimeria spp. remain largely unanswered. Here, we provide the broadest map yet of Eimeria occurrence for domestic chickens, confirming that all the known species (Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria mitis, Eimeria necatrix, Eimeria praecox, Eimeria tenella) are present in all six continents where chickens are found (including 21 countries). Analysis of 248 internal transcribed spacer sequences derived from 17 countries provided evidence of possible allopatric diversity for species such as E. tenella (FST values ⩽0.34) but not E. acervulina and E. mitis, and highlighted a trend towards widespread genetic variance. We found that three genetic variants described previously only in Australia and southern Africa (operational taxonomic units x, y and z) have a wide distribution across the southern, but not the northern hemisphere. While the drivers for such a polarised distribution of these operational taxonomic unit genotypes remains unclear, the occurrence of genetically variant Eimeria may pose a risk to food security and animal welfare in Europe and North America should these parasites spread to the northern hemisphere.
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http://dx.doi.org/10.1016/j.ijpara.2016.05.006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4978698PMC
August 2016

High-intensity cardiac infections of Phthinomita heinigerae n. sp. (Digenea: Aporocotylidae) in the orangelined cardinalfish, Taeniamia fucata (Cantor), off Heron Island on the Great Barrier Reef.

Parasitol Int 2016 Oct 14;65(5 Pt A):371-7. Epub 2016 May 14.

Centre for Sustainable Tropical Fisheries and Aquaculture, School of Marine and Tropical Biology, James Cook University, PO Box 6811, Cairns 4878, Australia; Fish Health Laboratory, Department of Fisheries Western Australia, 3 Baron-Hay Court, South Perth 6151, Australia. Electronic address:

We report a new species of aporocotylid trematode (Platyhelminthes: Digenea) from the heart of the orangelined cardinalfish, Taeniamia fucata (Cantor), from off Heron Island on the southern Great Barrier Reef. We used an integrated approach, analysing host distribution, morphology, and genetic data from the internal transcribed spacer 2 of the ribosomal DNA, to circumscribe Phthinomita heinigerae n. sp. This is the first species of Phthinomita Nolan & Cribb, 2006 reported from the Apogonidae; existing species and known 'types' are recorded from species of the Labridae, Mullidae, and Siganidae. The new species is distinguished from its 11 congeners in having a body 2977-3539 long and 16.5-22.4 times longer than wide, an anterior testis 6.2-8.2 times longer than wide and 8.3-13.0 times longer than the posterior testis, a posterior testis whose width is 35-56% of the body width, and an ovary positioned 11-13% of the body length from the posterior end, and is entirely anterior to the posterior margin of the anterior testis. In addition, 2-34 base differences (0.4-7.0% sequence divergence over 485 base positions) were detected among the ITS2 sequence representing P. heinigerae n. sp. and the 14 representing other Phthinomita species/molecular types. Prevalence and intensity of infection with P. heinigerae n. sp. was relatively high within the heart tissue of T. fucata, with 19 of 20 fish examined from off Heron Island infected (95%) with 7-25 adult worms (arithmetic mean 16.6). Infections by these parasites accounted for an occupation of 7-30% of the total estimated heart volume.
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http://dx.doi.org/10.1016/j.parint.2016.05.006DOI Listing
October 2016

Analysis of the function of IL-10 in chickens using specific neutralising antibodies and a sensitive capture ELISA.

Dev Comp Immunol 2016 10 22;63:206-12. Epub 2016 Apr 22.

The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK. Electronic address:

In mammals, the inducible cytokine interleukin 10 is a feedback negative regulator of inflammation. To determine the extent to which this function is conserved in birds, recombinant chicken IL-10 was expressed as a secreted human Ig Fc fusion protein (chIL-10-Fc) and used to immunise mice. Five monoclonal antibodies (mAb) which specifically recognise chicken IL-10 were generated and characterised. Two capture ELISA assays were developed which detected native chIL-10 secreted from chicken bone marrow-derived macrophages (chBMMs) stimulated with lipopolysaccharide (LPS). Three of the mAbs detected intracellular IL-10. This was detected in only a subset of the same LPS-stimulated chBMMs. The ELISA assay also detected massive increases in circulating IL-10 in chickens challenged with the coccidial parasite, Eimeria tenella. The same mAbs neutralised the bioactivity of recombinant chIL-10. The role of IL-10 in feedback control was tested in vitro. The neutralising antibodies prevented IL-10-induced inhibition of IFN-γ synthesis by mitogen-activated lymphocytes and increased nitric oxide production in LPS-stimulated chBMMs. The results confirm that IL-10 is an inducible feedback regulator of immune response in chickens, and could be the target for improved vaccine efficacy or breeding strategies.
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http://dx.doi.org/10.1016/j.dci.2016.04.016DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4947970PMC
October 2016

Molecular approaches to trematode systematics: 'best practice' and implications for future study.

Syst Parasitol 2016 Mar 22;93(3):295-306. Epub 2016 Feb 22.

Department of Pathology and Pathogen Biology, Royal Veterinary College, University of London, Hatfield, AL9 7TA, UK.

To date, morphological analysis has been the cornerstone to trematode systematics. However, since the late-1980s we have seen an increased integration of genetic data to overcome problems encountered when morphological data are considered in isolation. Here, we provide advice regarding the 'best molecular practice' for trematode taxonomy and systematic studies, in an attempt to help unify the field and provide a solid foundation to underpin future work. Emphasis is placed on defining the study goals and recommendations are made regarding sample preservation, extraction methods, and the submission of molecular vouchers. We advocate generating sequence data from all parasite species/host species/geographic location combinations and stress the importance of selecting two independently evolving loci (one ribosomal and one mitochondrial marker). We recommend that loci should be chosen to provide genetic variation suitable to address the question at hand and for which sufficient 'useful' comparative sequence data already exist. Quality control of the molecular data via using proof-reading Taq polymerase, sequencing PCR amplicons using both forward and reverse primers, ensuring that a minimum of 85% overlap exists when constructing consensus sequences, and checking electropherograms by eye is stressed. We advise that all genetic results are best interpreted using a holistic biological approach, which considers morphology, host identity, collection locality, and ecology. Finally, we consider what advances next-generation sequencing holds for trematode taxonomy and systematics.
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http://dx.doi.org/10.1007/s11230-016-9631-2DOI Listing
March 2016

Dollfustrema durum n. sp. and Heterobucephalopsis perardua n. sp. (Digenea: Bucephalidae) from the giant moray eel, Gymnothorax javanicus (Bleeker) (Anguilliformes: Muraenidae), and proposal of the Heterobucephalopsinae n. subfam.

Parasitol Int 2015 Dec 18;64(6):559-70. Epub 2015 Jul 18.

School of Biological Sciences, The University of Queensland, St Lucia, Queensland 4072, Australia. Electronic address:

Two new species of bucephalid trematode (Platyhelminthes: Digenea) are described from the giant moray eel, Gymnothorax javanicus (Anguilliformes: Muraenidae), from off Lizard Island, Australia. We used a combined morphological and molecular-based approach targeting the internal transcribed spacer 2 (ITS2) of the ribosomal DNA (rDNA) and the D1-D3 region of the large subunit (28S) of rDNA to circumscribe the species. Dollfustrema durum n. sp. is distinguished from seven congeners in having 5-6 rows of enlarged body spines circling the anterior portion of the rhynchus. From the remaining 10 species, D. durum n. sp. differs in body length, and in having a caecum that terminates posteriorly to the confluent arc formed by the vitelline follicles, gonads predominantly anterior to the pharynx, testes in tandem, an anterior testis positioned posteriorly to the vitelline follicles, and the pre-vitelline field 23-40% of the body length. Heterobucephalopsis perardua n. sp. differs from Heterobucephalopsis gymnothoracis, the type- and only other reported species, in being two to three times smaller. Heterobucephalopsis, currently considered a genus inquirendum, is confirmed as valid and is rediagnosed. Bayesian inference analysis of 28S rDNA sequences representing 28 species from nine genera and four subfamilies of bucephalid, indicates that i) subfamily classifications previously based on morphological characters are broadly robust, ii) the sequence representing H. perardua n. sp. is resolved as distinct, and basal, to sequences representing the Bucephalinae, the Prosorhynchinae, the Paurorhynchinae, and the Dolichoenterinae, iii) the Dolichoenterinae and the Prosorhynchinae are monophyletic sister clades, basal to the Bucephalinae and the Paurorhynchinae, iv) sequences representing Grammatorcynicola, Prosorhynchus, and Dollfustrema are also monophyletic, v) the Bucephalinae is paraphyletic relative to the Paurorhynchinae, and vi) the bucephaline genera Prosorhynchoides, Rhipidocotyle, and Bucephalus are each polyphyletic. The morphological and molecular differences observed among the four previously recognised subfamilies in this study lead us to propose Heterobucephalopsinae n. subfam. to accommodate the genus Heterobucephalopsis.
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http://dx.doi.org/10.1016/j.parint.2015.07.003DOI Listing
December 2015

Quantitative real-time PCR (qPCR) for Eimeria tenella replication--Implications for experimental refinement and animal welfare.

Parasitol Int 2015 Oct 2;64(5):464-70. Epub 2015 Jul 2.

Department of Pathology and Pathogen Biology, Royal Veterinary College, University of London, Hatfield AL9 7TA, United Kingdom. Electronic address:

The Eimeria species are highly pathogenic parasites of chickens. Research aimed at reducing their impact is hindered by a lack of non-subjective, quantitative, tools to measure parasite replication in the host. The time-consuming, and often time-sensitive, nature of existing approaches precludes their use in large-scale genetic, epidemiological, and evolutionary analyses. We have used quantitative real-time PCR (qPCR) to accurately quantify Eimeria tenella in chicken tissue and shown this to be more efficient and sensitive than traditional methodologies. We tested four chicken-specific reference qPCR assays and found beta-actin (actb) to be optimal for sample normalisation. In an experimental setting, chickens were inoculated with 500, 1500, or 4500 E. tenella oocysts and parasite replication and the impact of infection measured by i) qPCR analysis of DNA extracted from caecal tissues collected at five and eight days post-infection (dpi), ii) faecal oocyst counts (FOCs) on samples taken from six to eight dpi, and iii) lesion scoring on caeca collected post-mortem at five and eight dpi. Quantitative real-time PCR test results indicated a significant dose-dependent increase in parasite numbers among study groups for samples collected five dpi (i.e., prior to gametogony) (R(2)=0.994) (p<0.002) but not in those from day eight (after most oocyst shedding) (R(2)=0.006) (p>0.379). A strong dose-dependent increase in parasite replication and severity of infection was also revealed by FOC (R(2)=0.997) and lesion scoring. Importantly, qPCR offers substantial improvements for animal welfare via improved statistical power and reduced group sizes in experimental studies. The described qPCR method overcomes subjective limitations of coproscopic quantification, allows reproducible medium- to high-throughput examination of tissues, faeces, and oocysts, and is a valuable tool for determining the impact of Eimeria infections in both experimental and field settings.
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http://dx.doi.org/10.1016/j.parint.2015.06.010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4534708PMC
October 2015

Elucidating the life cycle of Marteilia sydneyi, the aetiological agent of QX disease in the Sydney rock oyster (Saccostrea glomerata).

Int J Parasitol 2015 May 9;45(6):419-26. Epub 2015 Mar 9.

Biodiversity Program, Queensland Museum, South Brisbane, Queensland 4101, Australia. Electronic address:

Marteilia sydneyi (Phylum Paramyxea, Class Marteiliidea, Order Marteiliida) (the causative agent of QX disease) is recognised as the most severe parasite to infect Saccostrea glomerata, the Sydney rock oyster, on the east coast of Australia. Despite its potential impact on industry (>95% mortality), research towards lessening these effects has been hindered by the lack of an experimental laboratory model of infection as a consequence of our incomplete understanding of the life cycle of this parasite. Here, we explored the presence of this parasite in hosts other than a bivalve mollusc from two study sites on the Hawkesbury River, New South Wales, Australia. We employed PCR-based in situ hybridisation and sequence analysis of a portion of the first internal transcribed spacer of rDNA in an attempt to detect M. sydneyi DNA in 21 species of polychaete worm. Marteilia DNA was detected in 6% of 1247 samples examined by PCR; the analysis of all amplicons defined one distinct sequence type for first internal transcribed spacer, representing M. sydneyi. Of the polychaete operational taxonomic units test-positive in PCR, we examined 116 samples via in situ hybridisation DNA probe staining and identified M. sydneyi DNA in the epithelium of the intestine of two specimens of Nephtys australiensis. Two differing morphological forms were identified: a 'primordial' cell that contained a well-defined nucleus but had little differentiation in the cytoplasm, and a 'plasmodial' cell that showed an apparent syncytial structure. This finding represents the first known record of the identification of M. sydneyi being parasitic in an organism other than an oyster, and only the third record of any species of Marteilia identified from non-molluscan hosts. Future work aims at determining if N. australiensis and S. glomerata are the only hosts in the life cycle of this paramyxean, and the development of experimental models to aid the production of QX disease-resistant oysters.
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http://dx.doi.org/10.1016/j.ijpara.2015.02.002DOI Listing
May 2015

The past, present, and future of Leishmania genomics and transcriptomics.

Trends Parasitol 2015 Mar 28;31(3):100-8. Epub 2015 Jan 28.

Dipartimento di Medicina Veterinaria, Università degli Studi di Bari, Bari, Italy.

It has been nearly 10 years since the completion of the first entire genome sequence of a Leishmania parasite. Genomic and transcriptomic analyses have advanced our understanding of the biology of Leishmania, and shed new light on the complex interactions occurring within the parasite-host-vector triangle. Here, we review these advances and examine potential avenues for translation of these discoveries into treatment and control programs. In addition, we argue for a strong need to explore how disease in dogs relates to that in humans, and how an improved understanding in line with the 'One Health' concept may open new avenues for the control of these devastating diseases.
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http://dx.doi.org/10.1016/j.pt.2014.12.012DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4356521PMC
March 2015

Cardicola beveridgei n. sp. (Digenea: Aporocotylidae) from the mangrove jack, Lutjanus argentimaculatus (Perciformes: Lutjanidae), and C. bullardi n. sp. from the Australian spotted mackerel, Scomberomorus munroi (Perciformes: Scombridae), from the northern Great Barrier Reef.

Parasitol Int 2014 Oct 27;63(5):735-45. Epub 2014 Jun 27.

School of Biological Sciences, The University of Queensland, St Lucia, Queensland 4072, Australia. Electronic address:

Cardicola Short, 1953 is a genus of the Aporocotylidae Odhner, 1912 (Digenea), with 25 currently recognised species described from 32 species of Perciformes and Mugiliformes fishes around the world, including eight species from the Great Barrier Reef. Here, we describe two new species from this region, namely Cardicola beveridgei n. sp. from the ventricle and atrium of the mangrove jack, Lutjanus argentimaculatus (Forsskål) (Perciformes: Lutjanidae), and Cardicola bullardi n. sp. from the ventricle of the Australian spotted mackerel, Scomberomorus munroi Collette & Russo (Perciformes: Scombridae), from off Lizard Island, Queensland, Australia. These two new species are most easily distinguished from the 25 current members of Cardicola in having the combination of i) a spinous oral sucker, ii) an anteriorly intercaecal ovary, iii) a uterus that extends anteriorly from the oötype, iv) the number of spines per ventrolateral transverse row, and in v) body size and the length/width ratio, vi) the oesophagus and caecal length(s) relative to body total length, vii) the length of the posterior caeca relative to the anterior pair, viii) the testis length/width ratio and its total size relative to that of the body, ix) the postovarian field as a percentage of body length, and x) egg size. In addition, C. beveridgei n. sp. is further differentiated by possessing a female genital pore that opens anterodextral to the male pore while C. bullardi n. sp. differs further in possessing a testis that is almost entirely intercaecal and does not extend anteriorly to the level of the intestinal bifurcation. Employing genetic analysis of ITS2 rDNA sequence data, representing these species and a further 13 recognised and three putative species of Cardicola, we were able to unequivocally confirm these specimens as distinct (9-22% different over 420 nucleotide positions). Distance analysis of ITS2 showed that i) species of Cardicola from the Siganidae formed a monophyletic clade, to the exclusion of other Cardicola species reported from the Scombridae, Sparidae, Lutjanidae and Chaetodontidae, ii) a general phylogenetic isolation exists between the species of Cardicola reported from scombrid fishes, and iii) C. beveridgei n. sp. and Cardicola milleri Nolan & Cribb, 2006 from lutjanids and Cardicola chaetodontis Yamaguti, 1970 from chaetodontids are phylogenetically close, despite the evolutionary remoteness between the host groups and their highly disparate biology. Given the likelihood of many additional species being attributed to Cardicola, we predict that continued molecular analyses will indicate that this genus will prove to incorporate a series of radiations in association with particular fish taxa as well as evidence of host-switching. (Nucleotide sequences reported in this paper are available in the GenBank database under accession no. KF752497).
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http://dx.doi.org/10.1016/j.parint.2014.06.006DOI Listing
October 2014

Impact of experimental hookworm infection on the human gut microbiota.

J Infect Dis 2014 Nov 3;210(9):1431-4. Epub 2014 May 3.

Australian Institute of Tropical Health and Medicine, Queensland Tropical Health Alliance Laboratory, James Cook University, Cairns.

The interactions between gastrointestinal parasitic helminths and commensal bacteria are likely to play a pivotal role in the establishment of host-parasite cross-talk, ultimately shaping the development of the intestinal immune system. However, little information is available on the impact of infections by gastrointestinal helminths on the bacterial communities inhabiting the human gut. We used 16S rRNA gene amplification and pyrosequencing to characterize, for the first time to our knowledge, the differences in composition and relative abundance of fecal microbial communities in human subjects prior to and following experimental infection with the blood-feeding intestinal hookworm, Necator americanus. Our data show that, although hookworm infection leads to a minor increase in microbial species richness, no detectable effect is observed on community structure, diversity or relative abundance of individual bacterial species.
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http://dx.doi.org/10.1093/infdis/jiu256DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4195438PMC
November 2014

RNA-Seq reveals infection-induced gene expression changes in the snail intermediate host of the carcinogenic liver fluke, Opisthorchis viverrini.

PLoS Negl Trop Dis 2014 Mar 27;8(3):e2765. Epub 2014 Mar 27.

Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia; Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom.

Background: Bithynia siamensis goniomphalos is the snail intermediate host of the liver fluke, Opisthorchis viverrini, the leading cause of cholangiocarcinoma (CCA) in the Greater Mekong sub-region of Thailand. Despite the severe public health impact of Opisthorchis-induced CCA, knowledge of the molecular interactions occurring between the parasite and its snail intermediate host is scant. The examination of differences in gene expression profiling between uninfected and O. viverrini-infected B. siamensis goniomphalos could provide clues on fundamental pathways involved in the regulation of snail-parasite interplay.

Methodology/principal Findings: Using high-throughput (Illumina) sequencing and extensive bioinformatic analyses, we characterized the transcriptomes of uninfected and O. viverrini-infected B. siamensis goniomphalos. Comparative analyses of gene expression profiling allowed the identification of 7,655 differentially expressed genes (DEGs), associated to 43 distinct biological pathways, including pathways associated with immune defense mechanisms against parasites. Amongst the DEGs with immune functions, transcripts encoding distinct proteases displayed the highest down-regulation in Bithynia specimens infected by O. viverrini; conversely, transcription of genes encoding heat-shock proteins and actins was significantly up-regulated in parasite-infected snails when compared to the uninfected counterparts.

Conclusions/significance: The present study lays the foundation for functional studies of genes and gene products potentially involved in immune-molecular mechanisms implicated in the ability of the parasite to successfully colonize its snail intermediate host. The annotated dataset provided herein represents a ready-to-use molecular resource for the discovery of molecular pathways underlying susceptibility and resistance mechanisms of B. siamensis goniomphalos to O. viverrini and for comparative analyses with pulmonate snail intermediate hosts of other platyhelminths including schistosomes.
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http://dx.doi.org/10.1371/journal.pntd.0002765DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3967946PMC
March 2014

Molecular-based investigation of Cryptosporidium and Giardia from animals in water catchments in southeastern Australia.

Water Res 2013 Apr 27;47(5):1726-40. Epub 2012 Dec 27.

Faculty of Veterinary Science, The University of Melbourne, Parkville, Victoria 3010, Australia.

There has been no large-scale systematic molecular epidemiological investigation of the waterborne protozoans, Cryptosporidium or Giardia, in southeastern Australia. Here, we explored, for the first time, the genetic composition of these genera in faecal samples from animals in nine Melbourne Water reservoir areas, collected over a period of two-years. We employed PCR-based single-strand conformation polymorphism (SSCP) and phylogenetic analyses of loci (pSSU and pgp60) in the small subunit (SSU) of ribosomal RNA and 60-kDa glycoprotein (gp60) genes to detect and characterise Cryptosporidium, and another locus (ptpi) in the triose-phosphate isomerase (tpi) gene to identify and characterise Giardia. Cryptosporidium was detected in 2.8% of the 2009 samples examined; the analysis of all amplicons defined 14 distinct sequence types for each of pSSU and pgp60, representing Cryptosporidium hominis (genotype Ib - subgenotype IbA10G2R2), Cryptosporidium parvum (genotype IIa - subgenotypes IIaA15G2R1, IIaA19G2R1, IIaA19G3R1, IIaA19G4R1, IIaA20G3R1, IIaA20G4R1, IIaA20G3R2 and IIaA21G3R1), Cryptosporidium cuniculus (genotype Vb - subgenotypes VbA22R4, VbA23R3, VbA24R3, VbA25R4 and VbA26R4), and Cryptosporidium canis, Cryptosporidium fayeri, Cryptosporidium macropodum and Cryptosporidium ubiquitum as well as six new pSSU sequence types. In addition, Giardia was identified in 3.4% of the samples; all 28 distinct ptpi sequence types defined were linked to assemblage A of Giardia duodenalis. Of all 56 sequence types characterised, eight and one have been recorded previously in Cryptosporidium and Giardia, respectively, from humans. In contrast, nothing is known about the zoonotic potential of 35 new genotypes of Cryptosporidium and Giardia recorded here for the first time. Future work aims to focus on estimating the prevalence of Cryptosporidium and Giardia genotypes in humans and a wide range of animals in Victoria and elsewhere in Australia. (Nucleotide sequences reported in this paper are available in the GenBank database under accession nos. KC282952-KC283005).
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http://dx.doi.org/10.1016/j.watres.2012.12.027DOI Listing
April 2013

Genetic characterisation of Cryptosporidium and Giardia from dairy calves: discovery of species/genotypes consistent with those found in humans.

Infect Genet Evol 2012 Dec 5;12(8):1984-93. Epub 2012 Sep 5.

Faculty of Veterinary Science, The University of Melbourne, Parkville, Victoria 3010, Australia.

Cryptosporidium and Giardia are important genera of parasitic protists that can cause significant diarrhoeal diseases in humans and other animals. Depending on the species/genotype of parasite, human infection may be acquired via anthroponotic or zoonotic transmission routes. Here, we undertook a molecular epidemiological investigation of these two genera of parasites in pre- and post-weaned calves from eight locations in Canterbury, New Zealand, by PCR-coupled sequencing and phylogenetic analysis of sequence data for regions in the 60 kDa glycoprotein (pgp60) gene of Cryptosporidium and/or the triose-phosphate isomerase (ptpi) gene of Giardia. The pgp60 and ptpi regions were specifically amplified from 15 (8.3%) and 11 (6.1%) of the 180 individual faecal samples, respectively. The sequences derived from all of the amplicons were aligned with homologous reference sequences and subjected to phylogenetic analysis by Bayesian inference. For Cryptosporidium, three samples contained Cryptosporidium parvum genotype IIa, subgenotypes IIaA15G3R1, IIaA19G3R1 and IIaA23G4. Twelve samples contained Cryptosporidium hominis genotype Ib, subgenotype IbA10G2R2. While subgenotypes IIaA15G3R1 and IIaA23G4 are new records, IIaA19G3R1 and IbA10G2R2 are commonly found in humans in various countries. For Giardia, two samples contained Giardia duodenalis assemblage A, also common in humans. In contrast, nine samples contained G. duodenalis assemblage E, which is the first report of this assemblage in cattle in New Zealand. Therefore, the present results indicate that dairy calves on the South Island of New Zealand harbour 'zoonotic' genotypes of Cryptosporidium and Giardia, which is likely to have significant public health implications.
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http://dx.doi.org/10.1016/j.meegid.2012.08.004DOI Listing
December 2012

First molecular characterization of Giardia duodenalis from goats in Malaysia.

Mol Cell Probes 2013 Feb 27;27(1):28-31. Epub 2012 Aug 27.

Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.

In the present study, 310 faecal samples from goats from eight different farms in Malaysia were tested for the presence of Giardia using a PCR-coupled approach. The nested PCR for SSU amplified products of the expected size (∼200 bp) from 21 of 310 (6.8%) samples. Sixteen of these 21 products could be sequenced successfully and represented six distinct sequence types. Phylogenetic analysis of the SSU sequence data using Bayesian Inference (BI) identified Giardia assemblages A, B and E. The identification of the 'zoonotic' assemblages A and B suggests that Giardia-infected goats represent a possible reservoir for human giardiasis in Malaysia.
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http://dx.doi.org/10.1016/j.mcp.2012.08.006DOI Listing
February 2013

Characterization of the eg95 gene family in the G6 genotype of Echinococcus granulosus.

Mol Biochem Parasitol 2012 Jun 18;183(2):115-21. Epub 2012 Feb 18.

The University of Melbourne, Werribee, Victoria 3030, Australia.

Cystic echinococcosis in humans and livestock animals is caused by infection with the cestode parasite Echinococcus granulosus. A number of genotypes of the parasite (designated G1-G10) are known to exist, with the genotype cluster G1-G3 and genotype G6 being responsible for the majority of humans infections. A recombinant vaccine has been developed for use in livestock to prevent infection with E. granulosus. The vaccine is based on the antigen EG95 which is expressed in the early larval stage (oncosphere) of the parasite. The EG95 antigen was originally cloned from the G1 genotype of E. granulosus and the protein has been found to be encoded by members of a small family of related genes in this genotype. Reliable information has not been available about the likely efficacy of the EG95 vaccine against genotypes other than G1. In this study, genomic DNA cloning techniques were used to characterize seven eg95-related gene fragments from the G6 genotype of E. granulosus. Three proteins appear to be encoded by these genes. Considerable differences were found between the EG95 related proteins from the G6 genotype compared with the EG95 protein from the G1 genotype. These differences suggest that the EG95-related proteins from the G6 genotype may have different antigenic epitopes compared with the current vaccine antigen. Data presented in this study have implications for future vaccine design and provide the information that would enable a G6 genotype-specific vaccine to be developed against E. granulosus, should this be considered a desirable addition to the available tools for control of cystic echinococcosis transmission.
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http://dx.doi.org/10.1016/j.molbiopara.2012.02.005DOI Listing
June 2012

Mutation scanning-based analysis of anisakid larvae from Sillago flindersi from Bass Strait, Australia.

Electrophoresis 2012 Feb;33(3):499-505

Department of Veterinary Science, The University of Melbourne, Werribee, Victoria, Australia.

Anisakidosis is an important fish-borne disease caused by the larvae of anisakid nematodes, which affects humans and a range of other animals. The accurate identification of members of this nematode group is central to investigating the epidemiology of the parasites and in the surveillance and control of anisakidosis. It is now well known that morphological identification alone does not allow specific identification, particularly of larval stages. To better understand the epidemiology of anisakid nematodes in southern Australian fishes and the potential risks posed to human health, a survey of 50 specimens of the commercially important fish, Sillago flindersi, from Bass Strait, Australia was conducted. We characterised anisakid larvae by PCR-coupled mutation scanning, sequencing and phylogenetic analyses of the first and second internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA. This study revealed that 92% of the S. flindersi examined were infected with anisakids (n=194), which were represented by seven genotypes. Phylogenetic analyses of the genotypes defined herein, together with reference sequence for Anisakis pegreffii and Hysterothylacium sp. from public databases (i.e. GenBank), revealed the presence of A. pegreffii (n=24), Hysterothylacium larval type IV (n=90) and Hysterothylacium larval type VIII (n=80) in S. flindersi. Thus, the PCR-coupled mutation scanning approach employed herein is an effective tool for the genetic characterisation of anisakid nematodes for diagnostic and analytical purposes (nucleotide sequences reported in this paper are available in the GenBank database under accession nos. JN631796-809).
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http://dx.doi.org/10.1002/elps.201100438DOI Listing
February 2012

Cryptic parasite revealed improved prospects for treatment and control of human cryptosporidiosis through advanced technologies.

Adv Parasitol 2011 ;77:141-73

Department of Veterinary Science, The University of Melbourne, Werribee, Victoria, Australia.

Cryptosporidium is an important genus of parasitic protozoa of humans and other vertebrates and is a major cause of intestinal disease globally. Unlike many common causes of infectious enteritis, there are no widely available, effective vaccine or drug-based intervention strategies for Cryptosporidium, and control is focused mainly on prevention. This approach is particularly deficient for infections of severely immunocompromised and/or suppressed, the elderly or malnourished people. However, cryptosporidiosis also presents a significant burden on immunocompetent individuals, and can, for example have lasting effects on the physical and mental development of children infected at an early age. In the last few decades, our understanding of Cryptosporidium has expanded significantly in numerous areas, including the parasite life-cycle, the processes of excystation, cellular invasion and reproduction, and the interplay between parasite and host. Nonetheless, despite extensive research, many aspects of the biology of Cryptosporidium remain unknown, and treatment and control are challenging. Here, we review the current state of knowledge of Cryptosporidium, with a focus on major advances arising from the recently completed genome sequences of the two species of greatest relevance in humans, namely Cryptosporidium hominis and Cryptosporidium parvum. In addition, we discuss the potential of next-generation sequencing technologies, new advances in in silico analyses and progress in in vitro culturing systems to bridge these gaps and to lead toward effective treatment and control of cryptosporidiosis.
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http://dx.doi.org/10.1016/B978-0-12-391429-3.00007-1DOI Listing
March 2012