Berrimah Veterinary Laboratories

Australia

Berrimah Veterinary Laboratories

Australia
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Milic N.L.,Charles Darwin University | Davis S.,Berrimah Veterinary Laboratories | Carr J.M.,Flinders University | Isberg S.,Charles Darwin University | And 3 more authors.
Developmental and Comparative Immunology | Year: 2015

A number of pathogens have been detected in crocodiles, however little is known about their ability to control these pathogens. The interferon stimulated gene (ISG), viperin, has gained attention recently as an important host protein involved in multiple arms of the immune response. Viperin in concert with a number of other ISGs was upregulated in response to viral nucleic acid mimics and sendai virus in the C. porosus cell line, LV-1, indicating an intact early innate response to viral infection in these animals for the first time. Viperin was cloned from the LV-1 cell line and shown to have similar localisation patterns as human viperin, as well as demonstrating extremely high conservation with the human orthologue, excepting at the N-terminus. Interestingly, C. porosus viperin was also able to inhibit Dengue virus replication in vitro, showing a high level of intact functionality for this protein across divergent animal species, and perhaps demonstrating its importance in the early innate response to pathogens in the animal kingdom. © 2015 Elsevier Ltd.


PubMed | Berrimah Veterinary Laboratories, University of Adelaide, Flinders University and Charles Darwin University
Type: Journal Article | Journal: Developmental and comparative immunology | Year: 2015

A number of pathogens have been detected in crocodiles, however little is known about their ability to control these pathogens. The interferon stimulated gene (ISG), viperin, has gained attention recently as an important host protein involved in multiple arms of the immune response. Viperin in concert with a number of other ISGs was upregulated in response to viral nucleic acid mimics and sendai virus in the C. porosus cell line, LV-1, indicating an intact early innate response to viral infection in these animals for the first time. Viperin was cloned from the LV-1 cell line and shown to have similar localisation patterns as human viperin, as well as demonstrating extremely high conservation with the human orthologue, excepting at the N-terminus. Interestingly, C. porosus viperin was also able to inhibit Dengue virus replication in vitro, showing a high level of intact functionality for this protein across divergent animal species, and perhaps demonstrating its importance in the early innate response to pathogens in the animal kingdom.


PubMed | University of Federal Defense Munich, Leibniz Institute for Zoo and Wildlife Research, Bielefeld University, Animal & Plant Health Agency and 2 more.
Type: Journal Article | Journal: PloS one | Year: 2016

The genus Brucella comprises various species of both veterinary and human medical importance. All species are genetically highly related to each other, sharing intra-species average nucleotide identities (ANI) of > 99%. Infections occur among various warm-blooded animal species, marine mammals, and humans. Until recently, amphibians had not been recognized as a host for Brucella. In this study, however, we show that novel Brucella species are distributed among exotic frogs worldwide. Comparative recA gene analysis of 36 frog isolates from various continents and different frog species revealed an unexpected high genetic diversity, not observed among classical Brucella species. In phylogenetic reconstructions the isolates consequently formed various clusters and grouped together with atypical more distantly related brucellae, like B. inopinata, strain BO2, and Australian isolates from rodents, some of which were isolated as human pathogens. Of one frog isolate (10RB9215) the genome sequence was determined. Comparative genome analysis of this isolate and the classical Brucella species revealed additional genetic material, absent from classical Brucella species but present in Ochrobactrum, the closest genetic neighbor of Brucella, and in other soil associated genera of the Alphaproteobacteria. The presence of gene clusters encoding for additional metabolic functions, flanked by tRNAs and mobile genetic elements, as well as by bacteriophages is suggestive for a different ecology compared to classical Brucella species. Furthermore it suggests that amphibian isolates may represent a link between free living soil saprophytes and the pathogenic Brucella with a preferred intracellular habitat. We therefore assume that brucellae from frogs have a reservoir in soil and, in contrast to classical brucellae, undergo extensive horizontal gene transfer.


Shilton C.,Berrimah Veterinary Laboratories | Brown G.P.,University of Sydney | Chambers L.,Berrimah Veterinary Laboratories | Benedict S.,Berrimah Veterinary Laboratories | And 4 more authors.
Veterinary Pathology | Year: 2014

Extremely poor growth of some individuals within a birth cohort (runting) is a significant problem in crocodile farming. We conducted a pathological investigation to determine if infectious disease is associated with runting in farmed saltwater crocodiles (Crocodylus porosus) and to look for evidence of other etiologies. In each of 2005 and 2007, 10 normal and 10 runt crocodiles, with an average age of 5.5 months and reared under identical conditions, were sampled. Laboratory testing included postmortem; histological examination of a wide variety of tissues (with quantitation of features that were noted subjectively to be different between groups); hematology; serum biochemistry (total protein, albumin, globulins, total calcium, phosphorus, and iron); bacterial culture of liver and spleen (2005 only); viral culture of liver, thymus, tonsil, and spleen using primary crocodile cell lines (2007 only); and serum corticosterone (2007 only). The only evidence of infectious disease was mild cutaneous poxvirus infection in 45% of normal and 40% of runt crocodiles and rare intestinal coccidia in 5% of normal and 15% of runt crocodiles. Bacterial and viral culture did not reveal significant differences between the 2 groups. However, runt crocodiles exhibited significant (P <.05) increases in adrenocortical cell cytoplasmic vacuolation and serum corticosterone, decreased production of bone (osteoporosis), and reduced lymphoid populations in the spleen, tonsil, and thymus. Runts also exhibited moderate anemia, hypoalbuminemia, and mild hypophosphatemia. Taken together, these findings suggest an association between runting and a chronic stress response (hyperactivity of the hypothalamic-pituitary-adrenal axis). © The Author(s) 2014.


Gauci P.J.,Defence Science and Technology Organisation, Australia | McAllister J.,Defence Science and Technology Organisation, Australia | Mitchell I.R.,Defence Science and Technology Organisation, Australia | Boyle D.B.,CSIRO | And 5 more authors.
PLoS ONE | Year: 2015

The Mapputta serogroup tentatively contains the mosquito-associated viruses Mapputta, Maprik, Trubanaman and Gan Gan. Interestingly, this serogroup has previously been associated with an acute epidemic polyarthritis-like illness in humans; however, there has been no ensuing genetic characterisation. Here we report the complete genome sequences of Mapputta and Maprik viruses, and a new Mapputta group candidate, Buffalo Creek virus, previously isolated from mosquitoes and detected by serology in a hospitalised patient. Phylogenetic analyses indicate that the group is one of the earliest diverged groups within the genus Orthobunyavirus of the family Bunyaviridae. Analyses show that these three viruses are related to the recently sequenced Australian bunyaviruses from mosquitoes, Salt Ash and Murrumbidgee. A notable feature of the Mapputta group viruses is the absence of the NSs (non-structural) ORF commonly found on the S segment of other orthobunyaviruses. Viruses of the Mapputta group have been isolated from geographically diverse regions ranging from tropical Papua New Guinea to the semi-arid climate of south-eastern Australia. The relevance of this group to human health in the region merits further investigation. © 2015 Gauci et al.


Hick P.,University of Sydney | Schipp G.,Berrimah Veterinary Laboratories | Bosmans J.,Darwin Aquaculture Center | Humphrey J.,Berrimah Veterinary Laboratories | Whittington R.,University of Sydney
Aquaculture | Year: 2011

Betanodavirus is a significant constraint to marine finfish aquaculture worldwide and the development of control strategies will depend on accurate data about mechanisms of infection. A repeated cross-sectional survey for betanodavirus infection using a real-time reverse transcriptase-polymerase chain reaction assay and virus isolation was conducted in barramundi (Lates calcarifer) at a hatchery in Australia to determine whether transmission occurred vertically from the broodstock or horizontally from the environment. Six consecutive production batches were studied in 2007-2008. Subclinical infection was detected in Batch 2007-1 with onset of infection after 26. d of age, reaching a true prevalence > 90.9% (lower 95% probability limit) by 40. d. Two days later an outbreak of VNN with 100% mortality occurred in 12. d old larvae in Batch 2007-2, due to infection with an identical betanodavirus, which was not detected in this batch 7. d previously. A point source epidemic initiated by horizontal transmission from the environment (probably seawater) to Batch 1 then Batch 2 was confirmed. Betanodavirus was not endemic in the hatchery and was probably not vertically transmitted because: (i) infection was not detected in the eggs or larvae of any batch; (ii) infection was not detected in 4 out of 6 batches; (iii) infection was not identified in 20 broodstock; iv) freedom from infection of life history stages was determined with a very high degree of confidence; v) similar data were derived from archival samples from a batch in 2005. The data suggested that outbreaks were initiated because of vulnerabilities in the water supply and spread because biosecurity measures failed. The age of fish appeared to be a major risk factor for susceptibility to disease. © 2011 Elsevier B.V.


Pizzatto L.,University of Sydney | Shilton C.M.,Berrimah Veterinary Laboratories | Shine R.,University of Sydney
Journal of Wildlife Diseases | Year: 2010

Host-parasite systems have often evolved over time, such that infection dynamics may become greatly modified from the time of initial contact of the host with the parasite. Biological invasions may be useful to clarify processes in the initial contact of hosts with parasites, and allow us to compare parasite uptake between the ancestral (coevolved) host and novel (noncoevolved) hosts. Cane toads (Bufo marinus) are spreading rapidly through tropical Australia, carrying with them a nematode lungworm (Rhabdias pseudosphaerocephala) congeneric with those found in Australian frogs. We investigated the dynamics of infections of the toad parasite by conducting histologic examinations of cane toads and three native Australian frogs (Litoria dahlii, Litoria nasuta, and Opisthodon ornatus) at 2, 6, and 10 days after experimental exposure to the toad lungworm. More worms were found in toads than in frogs, especially at longer periods postexposure. In toads, the infective larvae entered the skin and muscles within 2 days postexposure, passed into the coelom in 6 days, and reached the lungs at 10 days. In frogs, larvae were found in many organs rather than migrating to consistent target tissues; a few larvae reached the lungs of L. dahlii. Migratory larvae caused increasing inflammation (primarily granulomatous admixed with granulocytes then lymphocytes) through time, especially in frogs. Evolution has resulted in an enhanced ability of the lungworm to locate the target organ (the lungs) of the toad, and an increase in rates of parasite survival within this host. © Wildlife Disease Association 2010.


Brown G.P.,University of Sydney | Shilton C.M.,Berrimah Veterinary Laboratories | Shine R.,University of Sydney
Methods in Ecology and Evolution | Year: 2011

1. Measuring the degree of skin-swelling induced by intradermal injection of phytohemagglutinin (PHA) is simple, quick and inexpensive, does not require specialized equipment and is easily conducted under field conditions. 2.PHA is perhaps the most frequently used assay of immunocompetence in field studies of birds. However, the method has rarely been used, and never validated, for studies on ectothermic vertebrates. 3.Here, we document its use in an amphibian. In response to PHA injected into a toe web, cane toads exhibited a 35% increase in web thickness 24h postinjection. 4.Histologically, PHA injection initiated a rapid (<12h) infiltration of neutrophils, eosinophils and macrophages at the injection site, followed by an influx of lymphocytes by 24h postinjection. A second exposure to PHA stimulated a faster, more intense swelling response. 5.In cane toads, PHA injection elicits a rapid innate immune response, followed by a secondary response that may reflect cell-mediated immune activity. Both components are easily quantifiable by the degree of skin-swelling. 6.Hence, PHA injection offers a convenient assay to quantify immune function in anurans and could usefully be incorporated into studies on the reasons for global amphibian declines. © 2011 The Authors. Methods in Ecology and Evolution © 2011 British Ecological Society.


Nelson F.B.L.,University of Sydney | Brown G.P.,University of Sydney | Shilton C.,Berrimah Veterinary Laboratories | Shine R.,University of Sydney
International Journal for Parasitology: Parasites and Wildlife | Year: 2015

The cane toad invasion in Australia provides a robust opportunity to clarify the infection process in co-evolved versus de novo host-parasite interactions. We investigated these infection dynamics through histological examination following experimental infections of metamorphs of native frogs (Cyclorana australis) and cane toads (Rhinella marina) with Rhabdias hylae (the lungworm found in native frogs) and Rhabdias pseudosphaerocephala (the lungworm found in cane toads). Cane toads reared under continuous exposure to infective larvae of the frog lungworm were examined after periods of 2, 6, 10 and 15 days. Additionally, both toads and frogs were exposed for 24 h to larvae of either the toad or the frog lungworm, and examined 2, 5, 10 and 20 days post-treatment. R. hylae (frog) lungworms entered cane toads and migrated through the body but were not found in the target tissue, the lungs. Larvae of both lungworm species induced inflammation in both types of hosts, although the immune response (relative numbers of different cell types) differed between hosts and between parasite species. Co-evolution has modified the immune response elicited by infection and (perhaps for that reason) has enhanced the parasite's ability to survive and to reach the host's lungs. © 2015 The Authors.


Boyle D.B.,CSIRO | Amos-Ritchie R.,CSIRO | Broz I.,CSIRO | Walker P.J.,CSIRO | And 5 more authors.
Journal of Virology | Year: 2014

Bluetongue virus serotype 1 (BTV 1) was first isolated in Australia from cattle blood collected in 1979 at Beatrice Hill Farm (BHF), Northern Territory (NT). From long-term surveillance programs (1977 to 2011), 2,487 isolations of 10 BTV serotypes were made. The most frequently isolated serotype was BTV 1 (41%, 1,019) followed by BTV 16 (17.5%, 436) and BTV 20 (14%, 348). In 3 years, no BTVs were isolated, and in 12 years, no BTV 1 was isolated. Seventeen BTV 1 isolates were sequenced and analyzed in comparison with 10 Australian prototype serotypes. BTV 1 showed an episodic pattern of evolutionary change characterized by four distinct periods. Each period consisted primarily of slow genetic drift which was punctuated from time to time by genetic shifts generated by segment reassortment and the introduction of new genome segments. Evidence was found for coevolution of BTV genome segments. Evolutionary dynamics and selection pressure estimates showed strong temporal and clocklike molecular evolutionary dynamics of six Australian BTV genome segments. Bayesian coalescent estimates of mean substitution rates clustered in the range of 3.5×10-4 to 5.3×10-4 substitutions per site per year. All BTV genome segments evolved under strong purifying (negative) selection, with only three sites identified as under pervasive diversifying (positive) selection. The obligate replication in alternate hosts (insect vector and vertebrate hosts) imposed strong evolutionary constraints. The dominant mechanism generating genetic diversity of BTV 1 at BHF was through the introduction of new viruses and reassortment of genome segments with existing viruses. © 2014, American Society for Microbiology.

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