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Graphical abstract, Highlights • Biology and genetics suggest cryptic Eimeria Operational Taxonomic Units (OTUs) from chickens are new species. • New Eimeria spp. that infect chickens are pathogenic and require control. • Anticoccidial vaccination of chickens does not control three new Eimeria spp., More than 68 billion chickens were produced globally in 2018, emphasising their major contribution to the production of protein for human consumption and the importance of their pathogens. Protozoan Eimeria spp. are the most economically significant parasites of chickens, incurring global costs of more than UK £10.4 billion per annum. Seven Eimeria spp. have long been recognised to infect chickens, with three additional cryptic operational taxonomic units (OTUs) first described more than 10 years ago. As the world’s farmers attempt to reduce reliance on routine use of antimicrobials in livestock production, replacing drugs that target a wide range of microbes with precise species- and sometimes strain-specific vaccines, the breakthrough of cryptic genetic types can pose serious problems. Consideration of biological characteristics including oocyst morphology, pathology caused during infection and pre-patent periods, combined with gene-coding sequences predicted from draft genome sequence assemblies, suggest that all three of these cryptic Eimeria OTUs possess sufficient genetic and biological diversity to be considered as new and distinct species. The ability of these OTUs to compromise chicken bodyweight gain and escape immunity induced by current commercially available anticoccidial vaccines indicates that they could pose a notable threat to chicken health, welfare, and productivity. We suggest the names Eimeria lata n. sp., Eimeria nagambie n. sp. and Eimeria zaria n. sp. for OTUs x, y and z, respectively, reflecting their appearance (x) or the origins of the first isolates of these novel species (y, z).

Eimeria maxima is a common cause of coccidiosis in chickens, a disease that has a huge economic impact on poultry production. Knowledge of immunity to E. maxima and the specific mechanisms that contribute to differing levels of resistance observed between chicken breeds and between congenic lines derived from a single breed of chickens is required. This study aimed to define differences in the kinetics of the immune response of two inbred lines of White Leghorn chickens that exhibit differential resistance (line C.B12) or susceptibility (line 15I) to infection by E. maxima. Line C.B12 and 15I chickens were infected with E. maxima and transcriptome analysis of jejunal tissue was performed at 2, 4, 6 and 8 days post-infection (dpi). RNA-Seq analysis revealed differences in the rapidity and magnitude of cytokine transcription responses post-infection between the two lines. In particular, IFN-γ and IL-10 transcript expression increased in the jejunum earlier in line C.B12 (at 4 dpi) compared to line 15I (at 6 dpi). Line C.B12 chickens exhibited increases of IFNG and IL10 mRNA in the jejunum at 4 dpi, whereas in line 15I transcription was delayed but increased to a greater extent. RT-qPCR and ELISAs confirmed the results of the transcriptomic study. Higher serum IL-10 correlated strongly with higher E. maxima replication in line 15I compared to line C.B12 chickens. Overall, the findings suggest early induction of the IFN-γ and IL-10 responses, as well as immune-related genes including IL21 at 4 dpi identified by RNA-Seq, may be key to resistance to E. maxima.

Graphical abstract, Highlights • We measured tolerance and resistance to an Eimeria maxima infection of 2 broiler lines differing in their growth rate. • Infection induced typical symptoms of coccidiosis in relation to tolerance accompanied by parasite replication in the jejunum. • Lines did not differ in their resistance or tolerance. • Bone mineralisation was penalised by infection with effects being more pronounced at the recovery stage., 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

Eimeria maximais a common cause of coccidiosis in chickens, a disease which has a huge economic impact on poultry production. Knowledge of immunity toE. maximaand the specific mechanisms that contribute to differing levels of resistance observed between chicken breeds and between congenic lines derived from a single breed of chickens is required. This study aimed to define differences in the kinetics of the immune response of two inbred lines of White Leghorn chickens that exhibit differential resistance (line C.B12) or susceptibility (line 15I) to infection byE. maxima. Line C.B12 and 15I chickens were infected withE. maximaand transcriptome analysis of infected jejunal tissue was carried out at 2, 4, 6 and 8 days post-infection (dpi). RNA-Seq analysis revealed differences in the rapidity and magnitude of cytokine transcription responses post-infection between the two lines. In particular, IFN-γ and IL-10 transcripts in the jejunum accumulated earlier in line C.B12 (at 4 dpi) compared to line 15I (at 6 dpi). Line C.B12 chickens exhibited increases ofIFNGandIL10mRNA in the jejunum at 4 dpi, whereas in line 15I transcription was delayed but increased to a greater extent. RT-qPCR and ELISAs confirmed the results of the transcriptomic study. Higher serum IL-10 correlated strongly with higherE. maximareplication in line 15I compared to line C.B12 chickens. Overall, the findings suggest early induction of the IFN-γ and IL-10 responses, as well as immune-related genes at 4 dpi identified by RNA-Seq, may be key to resistance toE. maxima.

We surveyed 14 individuals of Tylosurus crocodilus Peron & 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.

Chlorotoxin was originally isolated from the venom of the Israeli scorpion Leiurus quinquestriatus, 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.

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.

Graphical abstract, Highlights • The seven Eimeria spp. recognised to infect chickens are present globally. • Cryptic Eimeria operational taxonomic units (OTUs) are common in the southern but not northern hemisphere. • Parasite population structure appears to vary between Eimeria spp., 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.

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.

Coccidiosis in poultry, caused by protozoan parasites of the genus Eimeria, 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 Eimeria parasitism. Prior infection with Eimeria maxima 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 E. maxima 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 E. maxima Houghton (H) strain, followed 3 weeks later by E. maxima Weybridge (W) strain challenge, while a backcross population received a single E. maxima 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 E. maxima 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).

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 Ruppell (Congridae); three additional species of Prosorhynchus are considered members of this genus, two of which are synonymised with S. uniporus.

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) (R2 = 0.994) (p 0.379). A strong dose-dependent increase in parasite replication and severity of infection was also revealed by FOC (R2 = 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., Graphical abstract, Highlights • Quantitative PCR indicates significant dose-dependent increases in parasites. • More sensitive measure of parasite replication than faecal oocyst count/lesion score • Significant implications for animal welfare and experiment refinement (3R's) • Support studies focusing on the genetic basis of resistance/susceptibility • Major implications for investigations of key aspects of Eimeria biology and control

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.

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.

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.

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. Electronic supplementary material The online version of this article (doi:10.1186/s13071-017-2283-5) contains supplementary material, which is available to authorized users.

International audience; The globally successful invasive snail, Potamopyrgus antipodarum (Tateidae), is rarely parasitized except in its native New Zealand where it is infected as first intermediate host by at least 20 species of trematode parasitic castrators. In France and Poland, only one trematode parasitic castrator, belonging to the Aporocotylidae (the blood flukes of fishes), is known to infect P. antipodarum, and its origin remains a mystery. We investigated the genetic identities, taxonomic relationships, and prevalences of this parasite and other aporocotylids infecting P. antipodarum and closely related snails in a French stream, and throughout New Zealand and southeast Australia. ITS2 DNA sequences of the French parasite matched with Aporocotylid sp. I (sensu Hechinger, 2012) from P. antipodarum in New Zealand. Hence, the aporocotylid parasite in Europe most likely originated from the native range of the introduced host. Aporocotylid sp. I was rare in both native and invasive ranges, infecting overall less than 1 out of 1,000 snails. The parasite and its host have a persistent relationship in France, as they both were repeatedly encountered over a period of 14 years. Our molecular phylogeny also reveals that this parasite is part of a diverse and poorly known group of aporocotylids in New Zealand and southeastern Australia.

The Great Barrier Reef holds the richest array of marine life found anywhere in Australia, including a diverse and fascinating parasite fauna. Members of one group, the trematodes, occur as sexually mature adult worms in almost all Great Barrier Reef bony fish species. Although the first reports of these parasites were made 100 years ago, the fauna has been studied systematically for only the last 25 years. When the fauna was last reviewed in 1994 there were 94 species known from the Great Barrier Reef and it was predicted that there might be 2,270 in total. There are now 326 species reported for the region, suggesting that we are in a much improved position to make an accurate prediction of true trematode richness. Here we review the current state of knowledge of the fauna and the ways in which our understanding of this fascinating group is changing. Our best estimate of the true richness is now a range, 1,100-1,800 species. However there remains considerable scope for even these figures to be incorrect given that fewer than one-third of the fish species of the region have been examined for trematodes. Our goal is a comprehensive characterisation of this fauna, and we outline what work needs to be done to achieve this and discuss whether this goal is practically achievable or philosophically justifiable.

$\textbf{BACKGROUND}$: 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. $\textit{Toxocara cati}$) exert on the gut microbiota of feline hosts. $\textbf{METHODS}$: The faecal microbiota of cats with patent infection by $\textit{T. cati}$ (= $\textit{Tc+}$), as well as that of negative controls (= $\textit{Tc-}$) 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. $\textbf{RESULTS}$: 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 $\textit{Enterococcaceae}$ and genera $\textit{Enterococcus}$ and $\textit{Dorea}$ showed a trend towards increased abundance in $\textit{Tc+}$ compared with $\textit{Tc-}$ samples, while no significant differences in OTU richness and diversity were recorded between $\textit{Tc+}$ and $\textit{Tc-}$ samples ($\textit{P}$=0.485 and $\textit{P}$=0.581, respectively). However, Canonical Correlation and Redundancy Analyses were able to separate samples by infection status ($\textit{P}$=0.030 and $\textit{P}$=0.015, respectively), which suggests a correlation between the latter and the composition of the feline faecal microbiota. $\textbf{CONCLUSIONS}$: In spite of the relatively small number of samples analysed, subtle differences in the composition of the gut microbiota of $\textit{Tc+}$ vs $\textit{Tc-}$ 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.

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.

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., Highlights • Five mouse mAbs to chicken IL-10 was produced. • Two capture ELISA assays were developed to quantify native IL-10. • Three mAbs (ROS-AV162, 163 and 164) neutralised the bioactivities of chIL-10. • Anti-IL-10 antibodies detected intracellular IL-10 in only a subset of LPS-stimulated macrophages. • Eimeria infection of chickens produced a massive increase in circulating IL-10.

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.

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).

We barcoded 25 in vitro isolates (representing 92 samples) of Giardia duodenalis from humans and other animals, which have been assembled by the Upcroft team at the Queensland Institute of Medical Research over a period of almost three decades. We used mutation scanning-coupled sequencing of loci in the triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh) and beta-giardin (bg) genes, combined with phylogenetic analysis, to genetically characterise them. Specifically, the isolates (n = 14) of G. duodenalis from humans from Australia (AD113; BRIS/83/HEPU/106; BRIS/87/HEPU/713; BRIS/89/HEPU/1003; BRIS/91/HEPU/1279; BRIS/92/HEPU/1541; BRIS/92/HEPU/1590; BRIS/92/HEPU/2443; BRIS/93/HEPU/1706), Malaysia (KL/92/IMR/1106) and Afghanistan (WB), a cat from Australia (BAC2), a sheep from Canada (OAS1) and a sulphur-crested cockatoo from Australia (BRIS/95/HEPU/2041) represented assemblage A (sub-assemblage AI-1, AI-2 or AII-2); isolates (n = 10) from humans from Australia (BRIS/91/HEPU/1279; BRIS/92/HEPU/2342; BRIS/92/HEPU/2348; BRIS/93/HEPU/1638; BRIS/93/HEPU/1653; BRIS/93/HEPU/1705; BRIS/93/HEPU/1718; BRIS/93/HEPU/1727), Papua New Guinea (BRIS/92/HEPU/1487) and Canada (H7) represented assemblage B (sub-assemblage BIV); and an isolate from cattle from Australia (BRIS/92/HEPU/1709) had a match to assemblage E. Isolate BRIS/90/HEPU/1229 from a human from Australia was shown to represent a mixed population of assemblages A and B. These barcoded isolates (including stocks and derived lines) now allow direct comparisons of experimental data among laboratories and represent a massive resource for transcriptomic, proteomic, metabolic and functional genomic studies using advanced molecular technologies. (Nucleotide sequences reported in this paper are available in the GenBank database under accession nos. XXXXX-XXXXX).

Polymerase chain reaction (PCR)-based single-strand conformation polymorphism (SSCP) and targeted sequencing were employed to genetically classify Echinococcus granulosus cysts from humans from 12 provinces in Mongolia using two DNA loci, designated pcox-1 and pnad-1, within the mitochondrial cytochrome c oxidase subunit 1 (cox-1) and NADH dehydrogenase subunit 1 (nad-1) genes, respectively. SSCP analysis of pcox-1 and pnad-1 amplicons produced from genomic DNA samples from individual E. granulosus cysts (n = 50) from individual humans displayed four distinct electrophoretic profiles for each pcox-1 and pnad-1. The direct sequencing of selected amplicons representing each of these profiles defined four distinct sequence types for each locus, present in four different combinations (designated as haplotypes M1-M4) for all 50 cyst isolates. Phylogenetic analysis of concatenated sequence data for these four haplotypes, including well-defined reference sequences, inferred that 68% of the cyst isolates belonged to the G1-G3 complex of E. granulosus (or E. granulosus sensu stricto), whereas the remaining (32%) were linked to the G6-G10 complex (or Echinococcus canadensis). Humans infected with E. granulosus cysts of the G1-G3 complex originated mainly from the eastern regions of Mongolia, whereas those harbouring cysts of the G6-G10 complex were from the western part of this country. The present study provides a first glimpse of the genetic composition of E. granulosus from humans in Mongolia, and forms a foundation for future studies of the epidemiology and ecology of the parasite(s) in animals and humans in this and surrounding countries.

In the United Kingdom, rabbits have been reported to harbour genotypes of Cryptosporidium (now recognized as C. cuniculus) identical to those from human patients exhibiting symptoms of cryptosporidiosis. The high density of rabbits in many regions of Australia, including both rural and urban as well as natural water catchments areas, and the absence of any information on Cryptosporidium from lagomorphs in this country stimulated the present study. We undertook an epidemiological study that genetically characterized Cryptosporidium from rabbits from four locations in Victoria by PCR-coupled sequencing and phylogenetic analysis of sequence data for loci within the small subunit of nuclear ribosomal RNA (SSU; for specific identification) and the 60kDa glycoprotein gene (gp60; for genotypic/subgenotypic identification). Cryptosporidium was detected in 12 (6.8%) of 176 individual faecal samples. For SSU, all 12 sequences were identical to each other and to that of C. cuniculus. For pgp60, all corresponding sequences matched the known genotype Vb, and were classified as subgenotype VbA23R3 (n=11) and VbA26R4 (n=1), which are both new records. Present evidence indicates that genotype Vb is limited to rabbits; however, it would be premature to conclude that this genotype is not zoonotic. Future studies should focus on the zoonotic potential of C. cuniculus from rabbits and a wide range of yet unstudied animals. (Nucleotide sequences reported in this paper are available in the GenBank database under accession nos. HM852431-HM852433).

Predicting how point mutations in genes alter the tertiary and quarternary structure of proteins is central to a number of areas of molecular biology and has implications in relation to the function and evolution of molecules. In the present study, we theoretically assessed the effects of 20 point mutations detected previously in a region of the triose-phosphate isomerase gene (tpi) of the protozoan Giardia duodenalis on the three-dimensional structure of the 'wild-type' protein (TPI). Amino acid substitutions arising from codon variations were mainly located at surface-accessible sites or in hydrophobic pockets of TPI. None of the substitutions was predicted to exert a significant change to the fold or functionality of the enzyme, with the exception of one alteration (Arg100). Almost all substitutions were either conservative or semi-conservative, and retained or even improved the expected stability of the fold. Overall, the findings provide support for the "neutral theory", which contends that evolution at the molecular level is not solely shaped by "Darwinian selection but also by random drift of selectively neutral or nearly neutral mutants".

Two new species of bucephalid trematodes are described from the rectum and intestine of the western moray eel Gymnothorax woodwardi McCulloch (Anguilliformes: Muraenidae: Muraeninae) off Point Peron in Western Australia. Dollfustrema gibsoni n. sp. is distinguished by body size, a pharynx that is intertesticular and level (latero-dextrally) with the anterior portion of the cirrus-sac, an ovary positioned dextrally to the testes and slightly anterior to (in part) the anterior testis, a uterus that extends anteriorly to the vitelline follicles but not to the level of the rhynchus, and vitelline follicles that form a confluent arc anterior to the gonads. Muraenicola nom. nov. is proposed as a replacement generic name for the pre-occupied Folliculovarium Gu & Shen 1983 nec Singh & Sinha 1981. Muraenicola botti n. sp. is distinguished from its congeners by body size, the size of the cirrus-sac (relative to body size), and in possessing tegumental spines, testes that are oblique (rather than in tandem) and eight ovarian lobes. It differs further in having an intestinal caecum that extends anteriorly to the level of the vitelline follicles and in the position of the pharynx and cirrus-sac relative to each other (lateral in part) as well as to the gonads. M. botti n. sp. also lacks a metraterm. These are the first reports of these genera from fishes off Australia and from the southern hemisphere.

This study explored the genetic make-up of Cryptosporidium in fecal samples from 268 individual calves on pasture-based dairy farms in three regions of Victoria, Australia. An integrated approach, using PCR-coupled single-strand conformation polymorphism, targeted sequencing and phylogenetic analysis, was employed to classify the genetic variants (i.e. genotypes and subgenotypes) of Cryptosporidium parvum present in 124 (46.3%) samples and to infer their zoonotic potential. Genotypic and subgenotypic classification was achieved using a portion of the 60 kDa glycoprotein gene (designated pgp60); specific identity was verified using a region within the small subunit of the nuclear ribosomal RNA (pSSU). Twelve sequence types representing ten distinct subgenotypes were defined within genotype IIa, namely IIaA16G3R1 (n=7), IIaA17G2R1 (1), IIaA18G2R1a (2), IIaA18G2R1b (1), IIaA18G4R1 (1), IIaA19G3R1a (80), IIaA19G3R1b (1), IIaA20G2R1 (9), IIaA20G3R1 (1), IIaA20G4R1 (9), IIaA21G3R1 (1) and IIaA23G3R1 (9), of which IIaA18G2R1b, IIaA18G4R1 and IIaA19G3R1b are new records. All of the subgenotypes, except IIaA16G3R1, IIaA18G4R1 and IIaA20G4R1, have been detected previously in humans and are thus considered to be of zoonotic relevance. (Nucleotide sequences reported in this paper are available in the GenBank database under accession numbers FJ825018-FJ825029).

A wide range of proteins belonging to the SCP/TAPS “family” has been described for various eukaryotic organisms, including plants and animals (vertebrates and invertebrates, such as helminths). Although SCP/TAPS proteins have been proposed to play key roles in a number of fundamental biological processes, such as host–pathogen interactions and defence mechanisms, there is a paucity of information on their genetic relationships, structures and functions, and there is no standardised nomenclature for these proteins. A detailed analysis of the relationships of members of the SCP/TAPS family of proteins, based on key protein signatures, could provide a foundation for investigating these areas. In this article, we review the current state of knowledge of key SCP/TAPS proteins of eukaryotes, with an emphasis on those from parasitic helminths, and undertake a comprehensive, systematic phylogenetic analysis of currently available full-length protein sequence data (considering characteristic protein signatures or motifs) to infer relationships and provide a framework (based on statistical support) for the naming of these proteins. This framework is intended to guide genomic and molecular biological explorations of key SCP/TAPS molecules associated with infectious diseases of plants and animals. In particular, fundamental investigations of these molecules in parasites and the integration of structural and functional data could lead to new and innovative approaches for the control of parasitic diseases, with important biotechnological outcomes.

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.

Pearsonellum pygmaeus n. sp. is described from Cromileptes altivelis (Serranidae), the Barramundi Cod, from Heron Island (southern Great Barrier Reef) and Lizard Island (northern Great Barrier Reef). This new species differs from Pearsonellum corventum (type and only species) in the combination of smaller overall body size, the relative distance of the brain from the anterior end, the relative lengths of both the oesophagus and the testis, the degree to which the testis extends outside the intercaecal field, the shape of the testis, the shape and size of the ovary and the extent to which the uterus loops around the ovary. There are in addition, 20 base pair differences between the ITS2 rDNA sequence of P. pygmaeus n. sp. and that of P. corventum. Three new host records for P. corventum are reported. Adelomyllos teenae n. g., n. sp. is described from Epinephelus coioides (Serranidae), the Estuary Cod, from Moreton Bay, southeast Queensland. The new genus differs from the 22 other sanguinicolid genera in the combined possession of two testes, a cirrus-sac, separate genital pores, a post-ovarian uterus and an H-shaped intestine. A. teenae n. sp. is the third sanguinicolid described from the Epinephelinae. Sanguinicolids have now been reported from 11 species of Serranidae.

The sanguinicolids Paracardicoloides yamagutii Martin, 1974 and Plethorchis acanthus Martin, 1975 were obtained from their definitive hosts, Anguilla reinhardtii Steindachner and Mugil cephalus Linnaeus (respectively) in the tributaries of the Brisbane River, Queensland, Australia. Two putative sanguinicolid cercariae were collected from a hydrobiid gastropod, Posticobia brazieri Smith, in the same waters. The two cercariae differ markedly in size and the form of their sporocysts. Both putative cercariae develop in the digestive gland of Po. brazieri. The ITS2 rDNA region from these sanguinicolids and a Clinostomum species (utilised as an outgroup due to the close morphological similarities between the cercarial stages of the Clinostomidae and the Sanguinicolidae) were sequenced and aligned. Comparison of the ITS2 sequences showed one cercaria to be that of P. yamagutii. This is the first sanguinicolid life history determined by a molecular method. P. yamagutii is the fourth sanguinicolid known to utilise a freshwater hydrobiid gastropod as its intermediate host. ITS2 rDNA is effective in distinguishing sanguinicolids at the species level.

Ankistromeces mariae n. g., n. sp. is described from Meuschenia freycineti (Monacanthidae), the six-spined leatherjacket, from off northern Tasmania. The new genus differs from the 21 other sanguinicolid genera in the combination of the anteriorly intercaecal and posteriorly post-caecal single testis, the presence of a cirrus-sac, the absence of an auxiliary external seminal vesicle, separate genital pores, the typically post-ovarian uterus and the H-shaped intestine. A. mariae is the first sanguinicolid to be reported from a monacanthid fish.

Highlights • Genomic and transcriptomic technologies are changing Leishmania research. • Research into new control strategies provides insights into the vector–parasite–host triangle. • Pathways of infection and disease in human and canine leishmaniases must be clarified., 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.

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.

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 (Forsskal) (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 ootype, 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).

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).

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.

5 páginas, 1 figura, 1 tabla, 22 figuras suplementarias, 16 tablas suplementarias -- PAGS nros. 1715-1719 et al., Wood is a major pool of organic carbon that is highly resistant to decay, owing largely to the presence of lignin. The only organisms capable of substantial lignin decay are white rot fungi in the Agaricomycetes, which also contains non–lignin-degrading brown rot and ectomycorrhizal species. Comparative analyses of 31 fungal genomes (12 generated for this study) suggest that lignin-degrading peroxidases expanded in the lineage leading to the ancestor of the Agaricomycetes, which is reconstructed as a white rot species, and then contracted in parallel lineages leading to brown rot and mycorrhizal species. Molecular clock analyses suggest that the origin of lignin degradation might have coincided with the sharp decrease in the rate of organic carbon burial around the end of the Carboniferous period, The work conducted by the U.S. Department of Energy Joint Genome Institute was supported by the Office of Science of the U.S. Department of Energy under contract DE-AC02-05CH11231. Also supported by the Assembling the Fungal Tree of Life (AFTOL) project under NSF awards DEB-0732968 (D.S.H.), DEB-0732993 (J.W.S.), and DEB-0732550 (D.J.M.).

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.

Infectious diarrhoeal diseases represent a major socio-economic burden to humans, and are linked to a range of pathogens, including viruses, bacteria and protists. The accurate detection of such pathogens is central to control. However, detection often relies on methods that have limited diagnostic sensitivity and specificity. Here, we assessed an automated, robotic platform for the simultaneous detection of eight major pathogens associated with infectious diarrhoea. Genomic DNA samples (n = 167) from faeces from humans with diarrhoea and diagnosed as cryptosporidiosis, and 100 uninfected control subjects, were tested for adenovirus 40/41, norovirus, Clostridium difficile, Campylobacter, Salmonella, Shigella, Cryptosporidium and Giardia by multiplexed-tandem PCR, and also characterized by single-strand conformation polymorphism analysis (SSCP) and selective sequencing. All 167 samples tested positive for Cryptosporidium, five for adenovirus 40/41, four for Campylobacter, three for C. difficile and seven for Shigella spp., with no false positive results for any assay. The automated PCR exhibited a high sensitivity, with10 individual pathogens being readily detected. The robotic detection platform assessed here represents a sensitive, high-throughput tool for key pathogens linked to infectious diarrhoea in humans. This platform requires little molecular biological expertise and is well suited to various diagnostic facilities and settings.