Melbourne Veterinary School - Research Publications

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    Measuring Sheep Tails: A Preliminary Study Using Length (Mm), Vulva Cover Assessment, and Number of Tail Joints
    Woodruff, M ; Munoz, C ; Coleman, G ; Doyle, R ; Barber, S (MDPI, 2023-03)
    Docking sheep tails is a long-standing practice that, when done at the recommended length, reduces the risk of flystrike. The recommended length is to cover the vulva of ewes and to a similar length in males. This length is often equated to three coccygeal joints left intact, and there are many other ways the recommended length is described by researchers, industry, and government. This study compared the observer consistency and retest consistency using three different tail length measurement methods: vulva cover assessment, length (mm), and joint palpation. The tails of 51 yearling and 48 weaner Merino ewes were assessed by two observers. Length and vulva cover assessment methods provided the most reliable results, and joint palpation was the least reliable method of tail measurement. In the sample, tails that covered the vulva of yearlings and weaners measured 57.6 mm (n = 14) and 63.7 mm (n = 30) on average, respectively, and contained two coccygeal joints (more than two coccygeal vertebrae). Tails that did not cover the vulva of yearlings and weaners measured 41.3 mm (n = 36) and 52.8 mm (n = 17) on average, respectively, and had less than two coccygeal joints. The two most reliable methods enable valid comparison to the best practice recommendations.
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    Immunoreactivity of canine, feline, and equine D-dimer with antibodies to human D-dimer
    Brown, JE ; Hadjinoormohammadi, A ; Courtman, NF (Wiley, 2024)
    BACKGROUND: Commercially available D-dimer assays use antibodies against human D-dimer, with limited sensitivity and specificity data in companion animals. OBJECTIVES: To evaluate the immunoreactivity of D-dimer in plasma of dogs, horses, and cats with commercially available antibodies to human D-dimer. ANIMALS: Plasma samples were collected from healthy dogs and horses, and from surplus feline plasma submitted for diagnostic purposes. METHODS: Descriptive research study. A cross-linked fibrin lysate was prepared from plasma samples, and SDS-PAGE and immunoblotting were performed with a variety of commercially available antibodies to human D-dimer. RESULTS: The selected antibodies demonstrated variable reactivity with D-dimer of each species. The monoclonal antibody DD44 bound canine D-dimer with good specificity and sensitivity, but this antibody did not react with feline or equine D-dimer. The polyclonal antibody D2D bound putative D-dimer in dogs, cats, and horses with good specificity, and higher sensitivity compared to human D-dimer. CONCLUSIONS AND CLINICAL IMPORTANCE: The variable performance of commercially available human D-dimer assays between species is, in part, because of inter-species variation in D-dimer immunoreactivity. The use of these assays should follow validation studies. Monoclonal antibody DD44 could be a focus for the development of a canine-specific assay.
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    Organic carbon utilisation by the filamentous alga Tribonema
    Liu, J ; Crosbie, ND ; Scales, PJ ; Martin, GJO (SPRINGER, 2024-01-01)
    Abstract Filamentous algae have potential application to wastewater treatment, in particular for efficient recovery of nutrients into biomass. However, supplying inorganic carbon is a major limiting factor. The utilisation of organic carbon present in wastewater may reduce the constraints in carbon supply, however there is little knowledge of mixotrophic growth amongst filamentous algae. This study investigated the utilisation of organic carbon sources relevant to wastewater by the filamentous xanthophyte alga Tribonema. Algae growth was compared in the absence of organic carbon (autotrophic) and in presence of 0.2 g-C L-1 glucose, ethanol or acetate under mixotrophic (presence of organic carbon and light) or heterotrophic (presence of organic carbon and absence of light) conditions. To investigate direct utilisation of organic carbon and indirect utilisation via bacterial CO2-genesis, cultivation was performed under both axenic and non-axenic conditions. Tribonema was found to directly utilise glucose, which increased mixotrophic productivity and maintained growth under heterotrophic conditions. In contrast, acetate was only indirectly utilised mixotrophically in the presence of bacteria, whereas ethanol was not utilised under any conditions. The underlying mechanisms of glucose utilisation by Tribonema were also investigated by analysing its photosynthetic rate and respiration rate under glucose concentrations ranging from 0 – 100 g L-1. Based on the results, enhancements to metabolic pathways and reduced CO2 requirements provided by glucose utilisation were proposed. Despite the positive results with respect to glucose utilisation, out competition for this resource by bacteria suggest that Tribonema is more suitable for treatment of wastewater with low organic carbon concentrations, such as secondary-treated wastewater effluent.
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    Evidence for dopamine production and distribution of dopamine D2 receptors in the equine gastrointestinal mucosa and pancreas
    Galinelli, NC ; Bamford, NJ ; de Laat, MA ; Sillence, MN ; Harris, PA ; Bailey, SR ; Li, H (PUBLIC LIBRARY SCIENCE, 2024-02-27)
    Insulin dysregulation in horses is characterised by hyperinsulinaemia and/or tissue insulin resistance and is associated with increased risk of laminitis. There is growing evidence in other species that dopamine attenuates insulin release from the pancreas; however, this has yet to be examined in horses. The present study aimed to identify whether there are cells capable of producing or responding to dopamine within the equine gastrointestinal mucosa and pancreas. Tissue samples were collected from the stomach, small and large intestines, and pancreas of six mature horses following euthanasia. Samples of stomach contents and faeces were also collected. Immunohistochemistry was performed to identify tyrosine hydroxylase (TH), the rate-limiting enzyme for dopamine production, and dopamine D2 receptors in tissue sections. Additional immunostaining for glucagon, insulin and chromogranin A was performed to identify α cells, β cells and enteroendocrine cells, respectively. Gastric parietal cells expressed both TH and D2 receptors, indicating that they are capable of both producing and responding to dopamine. Dopamine was quantified in stomach contents and faeces by high-performance liquid chromatography with electrochemical detection, with similar concentrations found at both sites. Dopamine D2 receptors were expressed in duodenal epithelial cells but not more distally. A subset of enteroendocrine cells, located sporadically along the gastrointestinal tract, were found to be immunopositive for the D2 receptor. In pancreatic islets, TH was present in α cells, while D2 receptors were strongly expressed in β cells and variably expressed in α cells. These findings are consistent with studies of other species; however, dynamic studies are required to further elucidate the role of dopamine in the modulation of insulin and glucagon secretion in horses. This descriptive study provides preliminary evidence for a potential role of dopamine to act as a paracrine messenger in the gastrointestinal mucosa and endocrine pancreas of horses.
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    Strengthening the role of innovation brokers in the livestock advisory services system of Pakistan
    Warriach, HM ; Ayre, M ; Nettle, R ; Height, K ; Iqbal, H ; Aziz, A ; Hayat, K ; Afzal, A ; Majeed, S ; Kumbher, A ; McGill, DM ; Bush, R (CSIRO Publishing, 2024)
    Context: This article assesses development and implementation of a non-traditional training ‘whole-family extension approach’ (WFEA) that contributes to the innovation-brokering capacity of farm advisors within the current livestock advisory services system of Pakistan. Aims: The primary objectives of the research were to explore how the WFEA training leads to improved capacity and knowledge of farm advisors and then examine these through the lens of innovation brokers and the difference they can make within the extension system, including (1) articulation of problems and possibilities, (2) network building and (3) supporting negotiation and learning in networks. Methods: A qualitative investigation was conducted during four facilitated discussions and three field follow-up visits following project interventions to build the capacity of 50 farm advisors from across a network of 22 organisations that are part of the Pakistani’s livestock extension system. Data were collected by using the following two qualitative approaches: (1) facilitated discussions, during each of the four separate training workshops; and (2) field follow-up visits, where farm advisors were interviewed following a set of semi-structured questions. The data were analysed to assess the changes observed across the different farm advisors during the training interventions and subsequent mentoring provided by the project team. Key results: Gaps in innovation-brokering capacity from WFEA were identified and include co-designing and more institutional support of various collaborating organisations, use of visioning tools, scenario analysis to predict possible future for the farm-advisor training and training of farm advisors on analytical skills to capture the household impacts; these should be incorporated in WFEA training. Conclusions: A holistic extension training intervention approach (the WFEA) can positively influence the innovation-brokering capacity of farm advisors within the current livestock advisory services system of Pakistan. Implications: This article has contributed to the literature on innovation-brokering roles in the livestock advisory systems by highlighting key additional functions of innovation-brokering in a developing-country context. Furthermore, examples from Pakistan can be used to show how farm advisors can develop skills in articulating problems and possibilities, network building and supporting negotiation and learning in networks.
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    Identification and treatment of Strongyloides stercoralis infection in a Boston Terrier dog from south-eastern Australia
    Chapman, SA ; Angles, JM ; Raw, C ; Zendejas-Heredia, PA ; Traub, RJ (Wiley, 2023)
    Strongyloides stercoralis, the causative agent of strongyloidiasis, is a potentially zoonotic intestinal nematode endemic to northern Australia. Strongyloidiasis is typically observed in immunocompromised hosts and is characterised by gastrointestinal signs, respiratory symptoms and a failure to thrive. In immunocompromised hosts, hyperinfection syndrome and disseminated infections can prove life-threatening. A 24-month-old Boston Terrier dog was referred for investigation of chronic small and large intestinal watery hematochezic diarrhoea, emaciation and hematemesis. Small intestinal histology identified a nematode despite consecutive negative faecal flotations. A real-time polymerase chain reaction and Baermann test subsequently confirmed infection with S. stercoralis. The dog had received an oral parasiticide comprising milbemycin oxime and afoxolaner every month for the 11 months prior to this diagnosis. Despite fenbendazole being reported as successful in the treatment of canine strongyloidiasis, a course of fenbendazole failed to clear the infection. Eradication of S. stercoralis infection was confirmed after the administration of off-label ivermectin fortnightly for 12 doses. Attention should be paid to this nematode as the failure of routine copromicroscopic methods to diagnose S. stercoralis infections can result in misdiagnosis, mistreatment and progression of the disease. Off-label ivermectin may be an alternative to fenbendazole for the treatment of Strongyloides spp. infection in dogs.
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    Genomic diversity and natural recombination of equid gammaherpesvirus 5 isolates
    Onasanya, AE ; El-Hage, C ; Diaz-Mendez, A ; Vaz, PK ; Legione, AR ; Devlin, JM ; Hartley, CA (ELSEVIER, 2023-11)
    BACKGROUND: Equid gammaherpesvirus 5 (EHV5) is closely related to equid gammaherpesvirus 2 (EHV2). Detection of EHV5 is frequent in horse populations worldwide, but it is often without a clear and significant clinical impact. Infection in horses can often present as subclinical disease; however, it has been associated with respiratory disease, including equine multinodular pulmonary fibrosis (EMPF). Genetic heterogeneity within small regions of the EHV5 glycoprotein B (gB) sequences have been reported and multiple genotypes of this virus have been identified within individual horses, but full genome sequence data for these viruses is limited. The primary focus of this study was to assess the genomic diversity and natural recombination among EHV5 isolates. RESULTS: The genome size of EHV5 prototype strain and the five EHV5 isolates cultured for this study, including four isolates from the same horse, ranged from 181,929 to 183,428 base pairs (bp), with the sizes of terminal repeat regions varying from 0 to 10 bp. The nucleotide sequence identity between the six EHV5 genomes ranged from 95.5 to 99.1%, and the estimated average nucleotide diversity between isolates was 1%. Individual genes displayed varying levels of nucleotide diversity that ranged from 0 to 19%. The analysis of nonsynonymous substitution (Ka > 0.025) revealed high diversity in eight genes. Genome analysis using RDP4 and SplitsTree programs detected evidence of past recombination events between EHV5 isolates. CONCLUSION: Genomic diversity and recombination hotspots were identified among EHV5 strains. Recombination can drive genetic diversity, particularly in viruses that have a low rate of nucleotide substitutions. Therefore, the results from this study suggest that recombination is an important contributing factor to EHV5 genomic diversity. The findings from this study provide additional insights into the genetic heterogeneity of the EHV5 genome.
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    Synthetic 5-amino-6-D-ribitylaminouracil paired with inflammatory stimuli facilitates MAIT cell expansion in vivo
    Nelson, AG ; Wang, H ; Dewar, PM ; Eddy, EM ; Li, S ; Lim, XY ; Patton, T ; Zhou, Y ; Pediongco, TJ ; Meehan, LJ ; Meehan, BS ; Mak, JYW ; Fairlie, DP ; Stent, AW ; Kjer-Nielsen, L ; Mccluskey, J ; Eckle, SBG ; Corbett, AJ ; Souter, MNT ; Chen, Z (FRONTIERS MEDIA SA, 2023-08-31)
    INTRODUCTION: Mucosal-associated invariant T (MAIT) cells are a population of innate-like T cells, which mediate host immunity to microbial infection by recognizing metabolite antigens derived from microbial riboflavin synthesis presented by the MHC-I-related protein 1 (MR1). Namely, the potent MAIT cell antigens, 5-(2-oxopropylideneamino)-6-D-ribitylaminouracil (5-OP-RU) and 5-(2-oxoethylideneamino)-6-D-ribitylaminouracil (5-OE-RU), form via the condensation of the riboflavin precursor 5-amino-6-D-ribitylaminouracil (5-A-RU) with the reactive carbonyl species (RCS) methylglyoxal (MG) and glyoxal (G), respectively. Although MAIT cells are abundant in humans, they are rare in mice, and increasing their abundance using expansion protocols with antigen and adjuvant has been shown to facilitate their study in mouse models of infection and disease. METHODS: Here, we outline three methods to increase the abundance of MAIT cells in C57BL/6 mice using a combination of inflammatory stimuli, 5-A-RU and MG. RESULTS: Our data demonstrate that the administration of synthetic 5-A-RU in combination with one of three different inflammatory stimuli is sufficient to increase the frequency and absolute numbers of MAIT cells in C57BL/6 mice. The resultant boosted MAIT cells are functional and can provide protection against a lethal infection of Legionella longbeachae. CONCLUSION: These results provide alternative methods for expanding MAIT cells with high doses of commercially available 5-A-RU (± MG) in the presence of various danger signals.
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    Development of a body condition index to estimate adiposity in ponies and horses from morphometric measurements
    Potter, SJJ ; Erdody, MLL ; Bamford, NJJ ; Knowles, EJJ ; Menzies-Gow, N ; Morrison, PKK ; Argo, CM ; McIntosh, BJJ ; Kaufman, K ; Harris, PAA ; Bailey, SRR (WILEY, 2024-03)
    BACKGROUND: There is a high prevalence of obesity in ponies and pleasure horses. This may be associated with equine metabolic syndrome and an increased risk of laminitis. Body condition scoring (BCS) systems are widely used but are subjective and not very sensitive. OBJECTIVES: To derive a body condition index (BCI), based on objective morphometric measurements, that correlates with % body fat. STUDY DESIGN: Retrospective cohort study. METHODS: Morphometric measurements were obtained from 21 ponies and horses in obese and moderate body condition. Percentage body fat was determined using the deuterium dilution method and the BCI was derived to give the optimal correlation with body fat, applying appropriate weightings. The index was then validated by assessing inter-observer variation and correlation with % body fat in a separate population of Welsh ponies; and finally, the correlation between BCI and BCS was evaluated in larger populations from studies undertaken in Australia, the United Kingdom and the United States. RESULTS: The BCI correlated well with adiposity in the ponies and horses, giving a Pearson r value of 0.74 (P < 0.001); however, it was found to slightly overestimate the % body fat in leaner animals and underestimate in more obese animals. In field studies, the correlation between BCI and BCS varied particularly in Shetlands and miniature ponies, presumably due to differences in body shape. MAIN LIMITATIONS: Further work may be required to adapt the BCI to a method that is more applicable for Shetlands and miniature ponies. CONCLUSIONS: This BCI was able to provide an index of adiposity which compared favourably with condition scoring in terms of accuracy of estimating adiposity; and was more consistent and repeatable when used by inexperienced assessors. Therefore, this may be a useful tool for assessing adiposity; and may be more sensitive than condition scoring for tracking weight gain or weight loss in individual animals.
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    Association between insulin dysregulation and adrenocorticotropic hormone in aged horses and ponies with no clinical signs of pituitary pars intermedia dysfunction
    Li, FI ; Spence, RJ ; de Laat, MA ; Harris, PA ; Sonntag, J ; Menzies-Gow, NJ ; Durham, AE ; Bailey, SR ; Sillence, MN (WILEY, 2023-11)
    BACKGROUND: High concentrations of adrenocorticotropic hormone (ACTH) are used as an indicator of pituitary pars intermedia dysfunction (PPID), but other factors that may influence ACTH need to be understood, if diagnostic reference ranges for ACTH are to be used with confidence. Insulin dysregulation (ID) could be one such factor, as insulin affects pituitary hormones in other species. OBJECTIVES: To test the hypothesis that a relationship exists between high insulin and high ACTH in aged (>15-year-old) animals with no clinical signs of PPID. STUDY DESIGN: A cohort study. METHODS: Thirteen horses and eleven ponies (17-25 years-old; mares and geldings) were clinically examined for signs of PPID in the spring (November 2020) and autumn (April 2021). On the same day, blood samples were taken before and 2 h after an oral glucose test (OGT). Concentrations of insulin, glucose, ACTH and cortisol were measured. RESULTS: There was no association between ACTH and cortisol. However, there was a positive linear correlation between ACTH and post-OGT (insulin in the autumn (r = 0.427, p = 0.04). Two horses and six ponies had ACTH above the cut-off value for PPID diagnosis, and of these eight animals, six also had insulin concentrations above the cut-off value for ID. MAIN LIMITATIONS: The cohort was small and thyrotropin-releasing hormone (TRH) stimulation tests were not performed. CONCLUSIONS: In autumn, high ACTH was associated with ID, when no clinical signs of PPID were present. Because ACTH is used in PPID diagnosis, further work is required to understand this interaction.