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The control of parasitic infections is particularly challenging in environments that are conducive to the maintenance of parasite lifecycles, such as the greyhound kennel, where the long-term breeding and rearing of dogs is common. The prevalence of gastrointestinal (GI) parasites within the Australian greyhound population has never previously been assessed, which seriously constrains the implementation of effective control measures. The aims of this study were to determine the prevalence and risk factors for GI parasites in Australian greyhounds, identify parasites which may be detrimental to the health and performance of dogs, and evaluate the likelihood of zoonotic transmission to kennel staff. Faecal samples were collected from 721 individual greyhounds situated in kennels across five states of Australia; Western Australia, Queensland, New South Wales, Victoria and Tasmania. Animal husbandry and current parasite control protocols were obtained from each kennel and analysed in conjunction with the detected level of parasitism. Overall parasite prevalence was approximately 60%, ranging from 50 to 70% between states. Eleven parasite genera were identified, with Sarcocystis, hookworm, Giardia and Toxocara detected most frequently. Generalised linear mixed model analyses found the major risk factors associated with parasitism were: a) the type of substrate which dogs were housed; b) age of dogs; and c) geographic region. Parasitism was associated most frequently with young dogs housed on grass/sand substrates, which allowed parasite lifecycles to continue, with constant reinfection the likely outcome. Routine treatment with broad-spectrum anthelmintics did not provide effective control in these environments and the adoption of alternate parasite control strategies is recommended. A substantial risk from zoonotic parasites was also identified, with six of the eleven parasite genera detected considered to be zoonotic and a poor understanding of zoonotic transmission among kennel managers.

During monitoring of critically endangered woylie (Bettongia penicillata) populations within the south-west of Western Australia, an adult female woylie was euthanased after being found in extremely poor body condition with diffuse alopecia, debilitating skin lesions and severe ectoparasite infestation. Trypanosoma copemani G2 and Sarcocystis sp. were detected molecularly within tissue samples collected post-mortem. Potorostrongylus woyliei and Paraustrostrongylus sp. nematodes were present within the stomach and small intestine, respectively. Blood collected ante-mortem revealed the presence of moderate hypomagnesaemia, mild hypokalaemia, mild hyperglobulinaemia and mild hypoalbuminaemia. Diffuse megakaryocytic hypoplasia was evident within the bone marrow. We propose various hypotheses that may explain the presence of severe ectoparasite infection, skin disease and poor body condition in this woylie. Given the potential deleterious effects of parasite infection, the importance of monitoring parasites cannot be over-emphasised., Graphical abstract Image 1, Highlights • Severe ectoparasite infestation, skin disease and poor body condition in a woylie. • Trypanosoma copemani genotype 2 and Sarcocystis sp. identified molecularly in tissues. • Clinical signs similar to those observed during the woylie decline.

Brushtail possums Trichosurus vulpecula remain in many areas of Perth, Western Australia, despite urbanisation. However, there are no data available regarding parasitic infections in this species in this locale, despite the relevance to wildlife health, and to public health when considering anthropozoonoses (infections that can spread from humans to animals, and vice versa). Further, though urbanisation is speculated to entail changes to wildlife infection epidemiology, there are few data investigating this hypothesis in marsupial populations in urbanised environments in Australia. This study aimed to measure T. gondii seroprevalence, gastrointestinal parasite prevalences, and macroscopic ectoparasite prevalences and intensities, in brushtail possums in the greater Perth region. It also aimed to compare infection prevalences between brushtail possum trapped in urbanised environments and bushland. As part of a cross-sectional study, 18 brushtail possums were trapped and sampled in bushland, 15 possums were trapped and sampled in urbanised environments, and 23 possum carcasses were obtained from a wildlife hospital, in the greater Perth region. This study provides parasite prevalence data, new host records for the ectoparasites Pygiopsylla tunneyi and Liponyssoides sp., and a new location record for the ectoparasite Haemaphysalis bremneri. Urbanised environments were inversely associated with prevalence of tick (Family Ixodidae) infections, and more specifically Amblyomma spp. infections. This study found no evidence that the Perth brushtail possum population is a substantial reservoir of anthropozoonotic parasites, though larger studies are required to confirm these findings.

New information generated since 2016 from the application of molecular tools to infections with Giardia and Cryptosporidium is critically summarised. In the context of molecular epidemiology, nomenclature, taxonomy, in vitro culture, detection, zoonoses, population genetics and pathogenicity, are covered. Whole genome sequencing has had the greatest impact in the last three years. Future advances will provide a much better understanding of the zoonotic potential of both parasites, their diversity and how this is linked to pathogenesis in different hosts.

Following confirmation that a remote village of approximately 300 inhabitants in northern Lao PDR was hyperendemic for the Neglected Tropical Disease Taenia solium, a pilot human-porcine therapeutic control intervention was implemented between October 2013 and November 2014. Mass drug administration with a three day albendazole 400mg protocol was offered to all eligible humans in October 2013 and March 2014. At these times, and again in October 2014, eligible village pigs received the anti-cysticercosis TSOL18 vaccination and an oral dose of oxfendazole anthelmintic at 30mg/kg, both repeated one month later. Community and individual human taeniasis prevalences were estimated via copro-antigen ELISA of volunteered human faecal samples prior to October 2013, and again in January 2015, in order to examine the short term impact of the intervention.

We aimed to validate the use of 1) the modified agglutination test and a polymerase chain reaction (PCR) protocol in detecting Toxoplasma gondii infection in quenda ( Isoodon obesulus) and brushtail possums ( Trichosurus vulpecula); 2) immunofluorescence microscopy of feces and a PCR and sequencing protocol in detecting Giardia spp. infection in quenda; and 3) a fecal flotation protocol in detecting gastrointestinal helminth infections of quenda. Quenda and brushtail possum carcasses, and samples from trapped quenda, were tested with 2 parasite detection tests per parasite, and results were modeled using Bayesian latent class analysis to estimate test sensitivity and specificity. The modified agglutination test and the PCR protocol were highly specific at detecting T. gondii infections in quenda and brushtail possums (≥93%); however, data were insufficient to assess sensitivity with adequate precision. Immunofluorescence microscopy and the PCR and sequencing protocol were both highly specific at detecting Giardia spp. in quenda (≥96%), but the PCR and sequencing protocol was relatively insensitive (69%, 95% credible interval [CrI]: 60–77%) compared to the highly sensitive immunofluorescence microscopy (98%, 95% CrI: 93–99%). The fecal flotation protocol was generally highly specific in the detection of gastrointestinal helminth infections (≥94%, with the exception of Trichuris spp. (88%, 95% CrI: 71–99%). The fecal flotation protocol was moderately to highly sensitive (≥74%) in the detection of strongyles, Labiobulura spp., Linstowinema spp., and Trichuris spp. Sensitivity was poor for detection of the cestode genus Potorolepis (36%, 95% CrI: 14–67%).

Giardia and Cryptosporidium are ubiquitous enteric protozoan pathogens of vertebrates. Although recognised as the aetiological agents of disease in humans and domestic animals for many years, fundamental questions concerning their ecology have been unresolved. Molecular tools have helped to better understand their genetic diversity and in so doing have helped to resolve questions about their transmission patterns and associated impacts on public health. However, the value of molecular tools is often complicated by questions concerning their applications, interpretation of results and terminology. Taxonomic issues have, until recently, made it difficult to determine the epidemiology of infections with both Giardia and Cryptosporidium. Similarly, improved understanding of their respective phylogenetic relationships has helped to resolve questions about zoonotic potential and distribution in wildlife. In the case of Cryptosporidium, imaging technologies have complemented phylogenetic studies in demonstrating the parasite's affinities with gregarine protozoa and have further supported its extracellular developmental capability and potential role as an environmental pathogen.

This report describes the detection and molecular characterisation of Neospora/Hammondia-like oocysts from naturally infected dogs. Single faecal samples were collected from 160 individual greyhounds and multiple faecal samples were collected from 5 Labrador litters within a breeding kennel. Microscopy analysis detected Neospora/Hammondia like oocysts in 2 greyhounds and one litter of Labrador puppies. PCR protocols targeting the internal transcribed spacer region (ITS-1) and the large subunit (LSU) rDNA (D2/D3 domains) confirmed the presence of H. heydorni in both greyhounds and N. caninum in the Labrador litter. Dietary information obtained for all positive dogs indicate the source of infection was through regular exposure to a commercially obtained raw meat diet. These results report for the first time a natural infection with H. heydorni in dogs in Australia and also the first co-occurrence of N. caninum and H. heydorni in an Australian urban region.

Limited knowledge regarding the biology and identification of the Australian tick Ixodes myrmecobii exists with only the female described to date. Here we provide a description of the male and nymph as well as a redescription of the female. All described stages are molecularly characterised using the cytochrome c oxidase subunit 1 (CO1) and internal transcribed spacer 2 (ITS2) loci. An updated list of hosts is presented including the first records from humans, cattle and several native species. Information on the distribution and conservation status of this species is also included.

Background: Individuals with Duchenne muscular dystrophy (DMD) often have lower intelligence than that of the general population. Furthermore, the prevalence of cognitive impairment, epilepsy, autism and attention deficit hyperactivity disorder (ADHD) are higher in DMD patients. Evidence suggests a role for the most predominant dystrophin isoform, Dp71, which is expressed in neuronal cells and glia. Aims: Due to limited knowledge of the brain of Duchenne patients and an unclear understanding of the function of Dp71, we sought to investigate how distal mutations in the DMD gene, which lead to an absence of Dp71 expression, are associated with the range of cognitive and behavioural phenotypes in these patients. We have modelled the loss of Dp71 by using DMD patient-derived fibroblasts (which naturally express the Dp71 isoform). We have also employed siRNA-mediated knockdown of Dp71 in a panel of neuronal and glial cell lines. We examined whether the loss of Dp71 was associated with defects in cell migration and proliferation given that alterations in these processes are associated with the cognitive and behavioural co-morbidities observed in DMD patients. Methods: Four patient-derived fibroblast cell lines, containing unique mutations of the DMD gene (resulting in the loss of Dp71) were examined using wound healing and proliferation assays as well as by RT-PCR and immunoblotting. The associated clinical data confirmed that patients exhibit cognitive and/or behavioural symptoms, which were severe for two individuals. Results: We demonstrate that DMD patient fibroblasts lacking Dp71 had reduced metabolic activity than control cells and migrated more rapidly. The increased level of cell migration is accompanied by an increase in focal adhesion kinase (FAK, a regulator of cell migration) expression. Conclusion: Our data highlights a role for Dp71 in cell migration and proliferation; alterations in these cellular processes are known to be associated with behavioural disorders prevalent in DMD patients such as autism. These findings have potential to inform the development of effective brain-targeting treatments.

Taxonomic identification of ticks obtained during a longitudinal survey of the critically endangered marsupial, Bettongia penicillata Gray, 1837 (woylie, brush-tailed bettong) revealed a new species of Ixodes Latrielle, 1795. Here we provide morphological data for the female and nymphal life stages of this novel species (Ixodes woyliei n. sp.), in combination with molecular characterisation using the mitochondrial cytochrome c oxidase subunit 1 gene (cox1). In addition, molecular characterisation was conducted on several described Ixodes species and used to provide phylogenetic context. Ixodes spp. ticks were collected from the two remaining indigenous B. penicillata populations in south-western Australia. Of 624 individual B. penicillata sampled, 290 (47%) were host to ticks of the genus Ixodes; specifically I. woyliei n. sp., I. australiensis Neumann, 1904, I. myrmecobii Roberts, 1962, I. tasmani Neumann, 1899 and I. fecialis Warburton & Nuttall, 1909. Of these, 123 (42%) were host to the newly described I. woyliei n. sp. In addition, 268 individuals from sympatric marsupial species (166 Trichosurus vulpecula hypoleucus Wagner, 1855 (brushtail possum), 89 Dasyurus geoffroii Gould, 1841 (Western quoll) and 13 Isoodon obesulus fusciventer Gray, 1841 (southern brown bandicoot)) were sampled for ectoparasites and of these, I. woyliei n. sp. was only found on two I. o. fusciventer. Morphological and molecular data have confirmed the first new Australian Ixodes tick species described in over 50 years, Ixodes woyliei n. sp. Based on the long-term data collected, it appears this tick has a strong predilection for B. penicillata, with 42% of Ixodes infections on this host identified as I. woyliei n. sp. The implications for this host-parasite relationship are unclear but there may be potential for a future co-extinction event. In addition, new molecular data have been generated for collected specimens of I. australiensis, I. tasmani and museum specimens of I. victoriensis Nuttall, 1916, which for the first time provides molecular support for the subgenus Endopalpiger Schulze, 1935 as initially defined. These genetic data provide essential information for future studies relying on genotyping for species identification or for those tackling the phylogenetic relationships of Australian Ixodes species.

Intelligence of individuals with Duchenne muscular dystrophy (DMD) is lower than the general population. Cognitive impairment, epilepsy, autism and attention deficit hyperactivity disorder are prevalent. Mounting evidence links these symptoms to the loss of dystrophin in the brain; yet central nervous system involvement in DMD has been largely ignored. The most predominant dystrophin isoform in the brain is the smaller Dp71 isoform expressed in neurons and glia. Distal DMD mutations affecting Dp71 expression are linked to cognitive impairment. The function(s) of Dp71 and how its absence can lead to cognitive and behavioural symptoms are not well understood. We are modelling the loss of Dp71 both endogenously in DMD patient-derived fibroblasts (which, in controls, naturally express Dp71) and with siRNA in neuronal and glial cell lines. Four patient fibroblast cell lines, each harbouring a different DMD gene mutation that naturally results in the loss of Dp71 are being studied. Accompanying clinical data confirms these patients display cognitive and/or behavioural symptoms, which is severe for two individuals. Our preliminary findings highlight a role for Dp71 in cell migration and proliferation; alterations in these processes are associated with psychiatric disorders prevalent in DMD such as autism. We will present would healing and proliferation assay data exploring these role(s) and discuss potential molecular mechanisms behind the cognitive and behavioural symptoms observed in DMD. The high risk of neuropsychiatric syndromes in Duchene warrants early intervention to achieve the best possible quality of life; our findings have significant potential to inform ongoing drug development in this area.

Calcium/strontium and 87Sr/86Sr ratios in foliage can be used to determine the relative importance of different soil sources of Ca to vegetation, if the discrimination of Ca/Sr by the plant between nutrient sources and foliage is known. We compared these tracers in the foliage of sugar maple (Acer saccharum) to the exchange fraction and acid leaches of soil horizons at six study sites in the White Mountains of New Hampshire, USA. In a previous study, sugar maple was shown to discriminate for Ca compared to Sr in foliage formation by a factor of 1.14 ± 0.12. After accounting for the predicted 14% shift in Ca/Sr, foliar Ca/Sr and 87Sr/86Sr ratios closely match the values in the Oie horizon at each study site across a 3.6-fold variation in foliar Ca/Sr ratios. Newly weathered cations, for which the Ca/Sr and 87Sr/86Sr ratios are estimated from acid leaches of soils, can be ruled out as a major Ca source to current foliage. Within sites, the 87Sr/86Sr ratio of the soil exchange pool in the Oa horizon and in the 0–10 cm and 10–20 cm increments of the mineral soil are similar to the Oie horizon and sugar maple foliar values, suggesting a common source of Sr in all of the actively cycling pools, but providing no help in distinguishing among them as sources to foliage. The Ca/Sr ratio in the soil exchange pool, however, decreases significantly with depth, and based on this variation, the exchange pool below the forest floor can be excluded as a major Ca source to the current sugar maple foliage. This study confirms that internal recycling of Ca between litter, organic soil horizons and vegetation dominate annual uptake of Ca in northern hardwood ecosystems. Refinement of our understanding of Ca and Sr uptake and allocation in trees allows improvement in the use of Ca/Sr and 87Sr/86Sr ratios to trace Ca sources to plants.

Following confirmation that a remote village of approximately 300 inhabitants in northern Lao PDR was hyperendemic for the Neglected Tropical Disease Taenia solium, a pilot human-porcine therapeutic control intervention was implemented between October 2013 and November 2014. Mass drug administration with a three day albendazole 400 mg protocol was offered to all eligible humans in October 2013 and March 2014. At these times, and again in October 2014, eligible village pigs received the anti-cysticercosis TSOL18 vaccination and an oral dose of oxfendazole anthelmintic at 30 mg/kg, both repeated one month later. Community and individual human taeniasis prevalences were estimated via copro-antigen ELISA of volunteered human faecal samples prior to October 2013, and again in January 2015, in order to examine the short term impact of the intervention.

Taenia solium taeniasis-cysticercosis and soil-transmitted helminths (STHs) are parasitic Neglected Tropical Diseases endemic throughout Southeast Asia. Within Lao PDR, a remote northern hill tribe village had previously been identified as a hyper endemic focus for T. solium. To reduce this observed prevalence, a One Health intervention covering both pigs and humans was implemented, which included two Mass drug administrations (MDA1 and MDA2) for village residents using a triple dose albendazole 400mg treatment regime. In addition to the effect on T. solium levels, the dual impact of this anthelmintic regime on STHs within the community was also monitored. Faecal samples were collected pre and post MDA1 and MDA2 and analysed for the presence of Taenia species and the STHs Ascaris lumbricoides, Trichuris trichiura and hookworm species. The McMaster technique was used to measure the changes in both prevalence and intensity of infection. Molecular characterisation of Taenia and hookworm species was conducted to detect zoonotic species. The level of taeniasis within the sampled population decreased by 79.4% after MDA1, remained steady during the five month inter-treatment interval and decreased again by 100% after MDA2. The prevalence of STHs decreased by 65.5% and 62.8% after MDA1 and MDA2 respectively; however an increase to 62.1% of pre MDA1 levels was detected during the inter-treatment interval. Individually, hookworm prevalence decreased by 83.4% (MDA1) and 84.5% (MDA2), A. lumbricoides by 95.6% and 93.5% and T. trichiura by 69.2% and 61%. The intensity of infection within the sampled population also decreased, with egg reduction rates of 94.4% and 97.8% for hookworm, 99.4% and 99.3% for A. lumbricoides and 77.2% and 88.5% for T. trichiura. Molecular characterisation identified a T. solium tapeworm carrier from 21.6% (13/60) of households in the village. T. saginata was identified in 5% (3/60) of households. The zoonotic hookworm A. ceylanicum was detected in the resident dog population. These results suggest that the triple dose albendazole 400mg treatment regime achieved a significant reduction in the level of taeniasis whilst simultaneously reducing the STH burden within the village. The increased STH prevalence detected between MDAs reflects the need for behavioural changes and a sustained chemotherapy programme, which may also need to include the resident dog population.

This case study discusses in detail for the first time the diagnosis and management of a case of leishmaniosis in a dog imported to Australia. The dog presented with epistaxis and a non-regenerative anaemia five years after being imported from Europe. Protozoa were identified within macrophages in bone marrow and splenic cytology. A Leishmania indirect fluorescent antibody test was performed and was positive while an Ehrlichia canis antibody test was negative. Polymerase chain reaction of the ITS-1 and ITS-2 regions of skin, lymph node, spleen and bone marrow were all positive for Leishmania infantum. The dog was treated with amphotericin B with a strong clinical response. The importance of thorough diagnostics in non-endemic areas, particularly Australia, is discussed. Treatment with amphotericin B is discussed. Vigilance, disease reporting and response frameworks are recommended for non-endemic areas.

Background Giardia-infection in cattle is often subclinical or asymptomatic, but it can also cause diarrhoea. The livestock-specific species Giardia bovis is the most frequently observed in cattle, however, the two zoonotic species Giardia duodenalis and Giardia enterica have also been found. Therefore calves are thought to be of public health significance. The aim of this study was to obtain current data about the frequency of the different Giardia-species in calves in Southern Germany. Findings Faecal samples of calves (diarrhoeic and healthy) in Southern Germany, diagnosed Giardia-positive by microscopy, were characterised by multi-locus PCR and sequencing. Of 152 microscopically Giardia-positive samples 110 (72.4%) were positive by PCR and successfully sequenced. G. bovis (Assemblage E) was detected in 101/110 (91.8%) PCR-positive samples, whilst G. duodenalis (Assemblage A) was detected in 8/110 (7.3%) samples and a mixed infection with G. duodenalis and G. bovis (Assemblage A+E) was identified in 1/110 (0.9%) samples. The sub-genotypes A1, E2 and E3 were identified with the β-giardin and the glutamate dehydrogenase genes. In the majority of diarrhoeic faecal samples a co-infection with Cryptosporidium spp. or Eimeria spp. was present, however, there were some in which G. bovis was the only protozoan pathogen found. Conclusions The results suggest that there is potentially a risk for animal handlers as calves in Southern Germany are, at a low percentage, infected with the zoonotic species G. duodenalis. In addition, it was found that G. bovis was the only pathogen identified in some samples of diarrhoeic calves, indicating that this parasite may be a contributing factor to diarrhoea in calves.

Within a remote Canadian Indigenous community, at least 11* of people had antibodies against Echinococcus granulosus and E. granulosus eggs were detected in 6* of environmentally collected canine fecal samples. Dog ownership, hunting, and trapping were not risk factors for seropositivity, suggesting that people are most likely exposed to E. granulosus through indirect contact with dog feces in the environment. In this situation, human exposure could be most effectively curtailed by preventing consumption of cervid viscera by free-roaming dogs.