Melbourne Veterinary School - Research Publications
Now showing items 1-12 of 109
Variation in the microbiome of the urogenital tract of Chlamydia-free female koalas (Phascolarctos cinereus) with and without 'wet bottom'
(PUBLIC LIBRARY SCIENCE, 2018-03-26)
Koalas (Phascolarctos cinereus) are iconic Australian marsupials currently threatened by several processes, including infectious diseases and ecological disruption. Infection with Chlamydia pecorum, is considered a key driver of population decline. The clinical sign of 'wet bottom', a staining of the rump associated with urinary incontinence, is often caused by chlamydial urinary tract infections. However, wet bottom has been recorded in koalas free of C. pecorum, suggesting other causative agents in those individuals. We used 16S rRNA diversity profiling to investigate the microbiome of the urogenital tract of ten female koalas in order to identify potential causative agents of wet bottom, other than C. pecorum. Five urogenital samples were processed from koalas presenting with wet bottom and five were clinically normal. All koalas were negative for C. pecorum infection. We detected thirteen phyla across the ten samples, with Firmicutes occurring at the highest relative abundance (77.6%). The order Lactobacillales, within the Firmicutes, comprised 70.3% of the reads from all samples. After normalising reads using DESeq2 and testing for significant differences (P < 0.05), there were 25 operational taxonomic units (OTUs) more commonly found in one group over the other. The families Aerococcaceae and Tissierellaceae both had four significantly differentially abundant OTUs. These four Tissierellaceae OTUs were all significantly more abundant in koalas with wet bottom. This study provides the foundation for future investigations of causes of koala wet bottom, other than C. pecorum infection. This is of clinical relevance as wet bottom is often assumed to be caused by C. pecorum and treated accordingly. Our research highlights that other organisms may be causing wet bottom, and these potential aetiological agents need to be further investigated to fully address the problems this species faces.
Immunoselected STRO-3(+) mesenchymal precursor cells reduce inflammation and improve clinical outcomes in a large animal model of monoarthritis
BACKGROUND: The purpose of this study was to investigate the therapeutic efficacy of intravenously administered immunoselected STRO-3 + mesenchymal precursor cells (MPCs) on clinical scores, joint pathology and cytokine production in an ovine model of monoarthritis. METHODS: Monoarthritis was established in 16 adult merino sheep by administration of bovine type II collagen into the left hock joint following initial sensitization to this antigen. After 24 h, sheep were administered either 150 million allogeneic ovine MPCs (n = 8) or saline (n = 8) intravenously (IV). Lameness, joint swelling and pain were monitored and blood samples for leukocytes and cytokine levels were collected at intervals following arthritis induction. Animals were necropsied 14 days after arthritis induction and gross and histopathological evaluations were undertaken on tissues from the arthritic (left) and contralateral (right) joints. RESULTS: MPC-treated sheep demonstrated significantly reduced clinical signs of lameness, joint pain and swelling compared with saline controls. They also showed decreased cartilage erosions, synovial stromal cell activation and angiogenesis. This was accompanied by decreased infiltration of the synovial tissues by CD4+ lymphocytes and CD14+ monocytes/macrophages. Over the 3 days following joint arthropathy induction, the numbers of neutrophils circulating in the blood and plasma concentrations of activin A were significantly reduced in animals administered MPCs. CONCLUSIONS: The results of this study have demonstrated the capacity of IV-administered MPCs to mitigate the clinical signs and some of the inflammatory mediators responsible for joint tissue destruction in a large animal model of monoarthritis.
Reducing Listening-Related Stress in School-Aged Children with Autism Spectrum Disorder
(SPRINGER/PLENUM PUBLISHERS, 2017-07-01)
High levels of stress and anxiety are common in children with Autism Spectrum Disorder (ASD). Within this study of school-aged children (20 male, 6 female) we hypothesised that functional hearing deficits (also pervasive in ASD) could be ameliorated by auditory interventions and that, as a consequence, stress levels would be reduced. The use of Ear-Level Remote Microphone devices and Classroom Amplification systems resulted in significantly improved listening, communication and social interaction and a reduction in physiologic stress levels (salivary cortisol) in both one-on-one and group listening situations.
Changes in BQCA Allosteric Modulation of [H-3]NMS Binding to Human Cortex within Schizophrenia and by Divalent Cations
(NATURE PUBLISHING GROUP, 2016-05-01)
Stimulation of the cortical muscarinic M1 receptor (CHRM1) is proposed as a treatment for schizophrenia, a hypothesis testable using CHRM1 allosteric modulators. Allosteric modulators have been shown to change the activity of CHRMs using cloned human CHRMs and CHRM knockout mice but not human CNS, a prerequisite for them working in humans. Here we show in vitro that BQCA, a positive allosteric CHRM1 modulator, brings about the expected change in affinity of the CHRM1 orthosteric site for acetylcholine in human cortex. Moreover, this effect of BQCA is reduced in the cortex of a subset of subjects with schizophrenia, separated into a discrete population because of a profound loss of cortical [(3)H]pirenzepine binding. Surprisingly, there was no change in [(3)H]NMS binding to the cortex from this subset or those with schizophrenia but without a marked loss of cortical CHRM1. Hence, we explored the nature of [(3)H]pirenzepine and [(3)H]NMS binding to human cortex and showed total [(3)H]pirenzepine and [(3)H]NMS binding was reduced by Zn(2+), acetylcholine displacement of [(3)H]NMS binding was enhanced by Mg(2+) and Zn(2+), acetylcholine displacement of [(3)H]pirenzepine was reduced by Mg(2+) and enhanced by Zn(2+), whereas BQCA effects on [(3)H]NMS, but not [(3)H]pirenzepine, binding was enhanced by Mg(2+) and Zn(2+). These data suggest the orthosteric and allosteric sites on CHRMs respond differently to divalent cations and the effects of allosteric modulation of the cortical CHRM1 is reduced in a subset of people with schizophrenia, a finding that may have ramifications for the use of CHRM1 allosteric modulators in the treatment of schizophrenia.
Changed cortical risk gene expression in major depression and shared changes in cortical gene expression between major depression and bipolar disorders
(SAGE PUBLICATIONS LTD, 2019-12-01)
BACKGROUND: Mood disorders likely occur in someone with a genetic predisposition who encounters a deleterious environmental factor leading to dysregulated physiological processes due to genetic mutations and epigenetic mechanisms altering gene expression. To gain data to support this hypothesis, we measured levels of gene expression in three cortical regions known to be affected by the pathophysiologies of major depression and bipolar disorders. METHODS: Levels of RNA were measured using the Affymetrix™ Human Exon 1.0 ST Array in Brodmann's areas 9, 10 and 33 (left hemisphere) from individuals with major depression, bipolar disorder and age- and sex-matched controls with changed expression taken as a fold change in RNA ⩾1.2 at p < 0.01. Data were analysed using JMP® genomics 6.0 and the probable biological consequences of changes in gene expression determined using Core and Pathway Designer Analyses in Ingenuity Pathway Analysis. RESULTS: There were altered levels of RNA in Brodmann's area 9 (major depression = 424; bipolar disorder = 331), Brodmann's area 10 (major depression = 52; bipolar disorder = 24) and Brodmann's area 33 (major depression = 59 genes; bipolar disorder = 38 genes) in mood disorders. No gene was differentially expressed in all three regions in either disorder. There was a high correlation between fold changes in levels of RNA from 112 genes in Brodmann's area 9 from major depression and bipolar disorder (r2 = 0.91, p < 0.001). Levels of RNA for four risk genes for major depression were lower in Brodmann's area 9 in that disorder. CONCLUSION: Our data argue that there are complex regional-specific changes in cortical gene expression in major depression and bipolar disorder that includes the expression of some risk genes for major depression in those with that disorder. It could be hypothesised that the common changes in gene expression in major depression and bipolar disorder are involved in the genesis of symptoms common to both disorders.
In Vitro Grown Pollen Tubes of Nicotiana alata Actively Synthesise a Fucosylated Xyloglucan
(PUBLIC LIBRARY SCIENCE, 2013-10-08)
Nicotiana alata pollen tubes are a widely used model for studies of polarized tip growth and cell wall synthesis in plants. To better understand these processes, RNA-Seq and de novo assembly methods were used to produce a transcriptome of N. alata pollen grains. Notable in the reconstructed transcriptome were sequences encoding proteins that are involved in the synthesis and remodelling of xyloglucan, a cell wall polysaccharide previously not thought to be deposited in Nicotiana pollen tube walls. Expression of several xyloglucan-related genes in actively growing pollen tubes was confirmed and xyloglucan epitopes were detected in the wall with carbohydrate-specific antibodies: the major xyloglucan oligosaccharides found in N. alata pollen grains and tubes were fucosylated, an unusual structure for the Solanaceae, the family to which Nicotiana belongs. Finally, carbohydrate linkages consistent with xyloglucan were identified chemically in the walls of N. alata pollen grains and pollen tubes grown in culture. The presence of a fucosylated xyloglucan in Nicotiana pollen tube walls was thus confirmed. The consequences of this discovery to models of pollen tube growth dynamics and more generally to polarised tip-growing cells in plants are discussed.
Assessment of serum symmetric dimethylarginine and creatinine concentrations in hyperthyroid cats before and after a fixed dose of orally administered radioiodine
Background: Serum symmetric dimethylarginine (SDMA) is a sensitive renal biomarker for detecting early chronic kidney disease (CKD) in nonhyperthyroid cats, but knowledge regarding its performance in hyperthyroid cats remains limited. Objectives: To determine the relationship between serum SDMA, creatinine and total thyroxine (TT4) concentrations in hyperthyroid cats before (T0) and 3 months after (T1) receiving a PO fixed dose of radioiodine. Animals: Eighty client‐owned hyperthyroid cats. Methods: Prospective cohort study. Serum TT4, and SDMA, creatinine concentrations, and urine specific gravity were measured at T0 and T1. Nonparametric tests were used to determine the relationship among SDMA, and creatinine and TT4 concentrations. Agreement between SDMA and creatinine regarding CKD staging at both time points was assessed using Goodman and Kruskal's gamma statistic. Results: Mean serum SDMA concentration increased after treatment of hyperthyroidism. However, 21 of 75 cats experienced a decrease in SDMA between T0 and T1, whereas creatinine decreased in only 2 cats. A moderate correlation between SDMA and creatinine was seen at T1 (r = 0.53; P < .001) but not at T0 (r = 0.13; P = .25). Where assessable at T1, poor agreement was observed between SDMA and creatinine and CKD stage (Goodman and Kruskal's gamma 0.20; P = .29). Conclusions and clinical importance: Discordant outcomes between SDMA and creatinine after radioiodine treatment in cats with hyperthyroidism suggest extrarenal factors may interfere with the reliability of SDMA to adequately reflect renal function. As a result, SDMA should not be interpreted in isolation in hyperthyroid cats treated with radioiodine.
Dafachronic acid promotes larval development in Haemonchus contortus by modulating dauer signalling and lipid metabolism
(PUBLIC LIBRARY SCIENCE, 2019-07-01)
Here, we discovered an endogenous dafachronic acid (DA) in the socioeconomically important parasitic nematode Haemonchus contortus. We demonstrate that DA promotes larval exsheathment and development in this nematode via a relatively conserved nuclear hormone receptor (DAF-12). This stimulatory effect is dose- and time-dependent, and relates to a modulation of dauer-like signalling, and glycerolipid and glycerophospholipid metabolism, likely via a negative feedback loop. Specific chemical inhibition of DAF-9 (cytochrome P450) was shown to significantly reduce the amount of endogenous DA in H. contortus; compromise both larval exsheathment and development in vitro; and modulate lipid metabolism. Taken together, this evidence shows that DA plays a key functional role in the developmental transition from the free-living to the parasitic stage of H. contortus by modulating the dauer-like signalling pathway and lipid metabolism. Understanding the intricacies of the DA-DAF-12 system and associated networks in H. contortus and related parasitic nematodes could pave the way to new, nematode-specific treatments.
Genomic recombination between infectious laryngotracheitis vaccine strains occurs under a broad range of infection conditions in vitro and in ovo
(PUBLIC LIBRARY SCIENCE, 2020-03-02)
Gallid alphaherpesvirus 1 causes infectious laryngotracheitis (ILT) in farmed poultry worldwide. Intertypic recombination between vaccine strains of this virus has generated novel and virulent isolates in field conditions. In this study, in vitro and in ovo systems were co-infected and superinfected under different conditions with two genomically distinct and commonly used ILTV vaccines. The progeny virus populations were examined for the frequency and pattern of recombination events using multi-locus high-resolution melting curve analysis of polymerase chain reaction products. A varied level of recombination (0 to 58.9%) was detected, depending on the infection system (in ovo or in vitro), viral load, the composition of the inoculum mixture, and the timing and order of infection. Full genome analysis of selected recombinants with different in vitro phenotypes identified alterations in coding and non-coding regions. The ability of ILTV vaccines to maintain their capacity to recombine under such varied conditions highlights the significance of recombination in the evolution of this virus and demonstrates the capacity of ILTV vaccines to play a role in the emergence of recombinant viruses.
Epidermal cell death in frogs with chytridiomycosis
(PEERJ INC, 2017-02-01)
BACKGROUND: Amphibians are declining at an alarming rate, and one of the major causes of decline is the infectious disease chytridiomycosis. Parasitic fungal sporangia occur within epidermal cells causing epidermal disruption, but these changes have not been well characterised. Apoptosis (planned cell death) can be a damaging response to the host but may alternatively be a mechanism of pathogen removal for some intracellular infections. METHODS: In this study we experimentally infected two endangered amphibian species Pseudophryne corroboree and Litoria verreauxii alpina with the causal agent of chytridiomycosis. We quantified cell death in the epidermis through two assays: terminal transferase-mediated dUTP nick end-labelling (TUNEL) and caspase 3/7. RESULTS: Cell death was positively associated with infection load and morbidity of clinically infected animals. In infected amphibians, TUNEL positive cells were concentrated in epidermal layers, correlating to the localisation of infection within the skin. Caspase activity was stable and low in early infection, where pathogen loads were light but increasing. In animals that recovered from infection, caspase activity gradually returned to normal as the infection cleared. Whereas, in amphibians that did not recover, caspase activity increased dramatically when infection loads peaked. DISCUSSION: Increased cell death may be a pathology of the fungal parasite, likely contributing to loss of skin homeostatic functions, but it is also possible that apoptosis suppression may be used initially by the pathogen to help establish infection. Further research should explore the specific mechanisms of cell death and more specifically apoptosis regulation during fungal infection.
Field and Laboratory Studies of the Susceptibility of the Green Treefrog (Hyla cinerea) to Batrachochytrium dendrobatidis Infection
(PUBLIC LIBRARY SCIENCE, 2012-06-07)
Amphibians worldwide are experiencing devastating declines, some of which are due to the amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd). Populations in the southeastern United States, however, have not been noticeably affected by the pathogen. The green treefrog (Hyla cinerea) is abundant and widespread in the southeastern United States, but has not been documented to harbor Bd infection. This study examined the susceptibility of H. cinerea to two strains of Bd in the lab and the prevalence of infection in wild populations of this species in southeastern Louisiana. Although we were able to infect H. cinerea with Bd in the lab, we did not observe any clinical signs of chytridiomycosis. Furthermore, infection by Bd does not appear to negatively affect body condition or growth rate of post-metamorphic individuals. We found no evidence of infection in surveys of wild H. cinerea. Our results suggest that H. cinerea is not susceptible to chytridiomycosis post-metamorphosis and probably is not an important carrier of the fungal pathogen Bd in the southeastern United States, although susceptibility at the larval stage remains unknown.
Fungal infection has sublethal effects in a lowland subtropical amphibian population
BACKGROUND: The amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), has been implicated as a primary cause of decline in many species around the globe. However, there are some species and populations that are known to become infected in the wild, yet declines have not been observed. Here we conducted a yearlong capture-mark-recapture study and a 2-year long disease monitoring study of northern cricket frogs, Acris crepitans, in the lowland subtropical forests of Louisiana. RESULTS: We found little evidence for an impact of Bd infection on survival; however, Bd infection did appear to cause sublethal effects, including increased capture probability in the field. CONCLUSIONS: Our study suggests that even in apparently stable populations, where Bd does not appear to cause mortality, there may be sublethal effects of infection that can impact a host population's dynamics and structure. Understanding and documenting such sublethal effects of infection on wild, seemingly stable populations is important, particularly for predicting future population declines.