Veterinary Biosciences - Research Publications

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    Infectious Laryngotracheitis Virus Viral Chemokine-Binding Protein Glycoprotein G Alters Transcription of Key Inflammatory Mediators In Vitro and In Vivo
    Coppo, MJC ; Devlin, JM ; Legione, AR ; Vaz, PK ; Lee, S-W ; Quinteros, JA ; Gilkerson, JR ; Ficorilli, N ; Reading, PC ; Noormohammadi, AH ; Hartley, CA ; Sandri-Goldin, RM (AMER SOC MICROBIOLOGY, 2018-01)
    Infectious laryngotracheitis virus (ILTV) is an alphaherpesvirus that infects chickens, causing upper respiratory tract disease and significant losses to poultry industries worldwide. Glycoprotein G (gG) is a broad-range viral chemokine-binding protein conserved among most alphaherpesviruses, including ILTV. A number of studies comparing the immunological parameters between infection with gG-expressing and gG-deficient ILTV strains have demonstrated that expression of gG is associated with increased virulence, modification of the amount and the composition of the inflammatory response, and modulation of the immune responses toward antibody production and away from cell-mediated immune responses. The aims of the current study were to examine the establishment of infection and inflammation by ILTV and determine how gG influences that response to infection. In vitro infection studies using tracheal organ tissue specimen cultures and blood-derived monocytes and in vivo infection studies in specific-pathogen-free chickens showed that leukocyte recruitment to the site of infection is an important component of the induced pathology and that this is influenced by the expression of ILTV gG and changes in the transcription of the chicken orthologues of mammalian CXC chemokine ligand 8 (CXCL8), chicken CXCLi1 and chicken CXCLi2, among other cytokines and chemokines. The results from this study demonstrate that ILTV gG interferes with chemokine and cytokine transcription at different steps of the inflammatory cascade, thus altering inflammation, virulence, and the balance of the immune response to infection.IMPORTANCE Infectious laryngotracheitis virus is an alphaherpesvirus that expresses gG, a conserved broad-range viral chemokine-binding protein known to interfere with host immune responses. However, little is known about how gG modifies virulence and influences the inflammatory signaling cascade associated with infection. Here, data from in vitro and in vivo infection studies are presented. These data show that gG has a direct impact on the transcription of cytokines and chemokine ligands in vitro (such as chicken CXCL8 orthologues, among others), which explains the altered balance of the inflammatory response that is associated with gG during ILTV infection of the upper respiratory tract of chickens. This is the first report to associate gG with the dysregulation of cytokine transcription at different stages of the inflammatory cascade triggered by ILTV infection of the natural host.
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    Mapping B lymphocytes as major reservoirs of naturally occurring latent equine herpesvirus 5 infection
    Mekuria, ZH ; El-Hage, C ; Ficorilli, NP ; Washington, EA ; Gilkerson, JR ; Hartley, CA (MICROBIOLOGY SOC, 2017-03)
    Equine herpesvirus 5 (EHV5) is a commonly detected gammaherpesvirus, which, along with the closely related EHV2, constitute the only two known percaviruses that infect horses. Apart from detection in horse populations worldwide and the recent publication of the whole genome, there is little known about the biology and pathogenesis of this virus, with many assumptions made by parallels with EHV2. The long-term survival of gammaherpesviruses within infected hosts involves the establishment and maintenance of latency in selected cell and tissues types, particularly lymphocytes. A latent gammaherpesvirus infection is characterized by a limited number of genes expressing in a particular cell or tissue type. In this study, we have used in vitro co-culturing to detect EHV5 in equine PBMCs and characterize the predominant cellular site for the establishment and maintenance of a latent infection. These experiments were conducted by isolating PBMCs from 10 horses and sorting subpopulations into two T lymphocyte (CD4 and CD8), B lymphocyte and macrophage enriched or depleted fractions. These lymphocyte and macrophage fractions were examined for the presence of latent EHV5 by in vitro co-culturing with equine foetal kidney cells. The lymphocyte fraction enriched with B lymphocytes had a significantly increased (P=0.005) number of plaques formed during co-culturing, whereas the B lymphocyte depleted fraction had a significant reduction in the number of plaques formed after co-culturing. Taken together, these results demonstrate that equine gammaherpesviruses establish latency in the equine PBMCs, with the predominant site for maintenance of latent virus being B lymphocytes.
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    The use of social network analysis to examine the transmission of Salmonella spp. within a vertically integrated broiler enterprise
    Crabb, HK ; Allen, JL ; Devlin, JM ; Firestone, SM ; Stevenson, MA ; Gilkerson, JR (ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD, 2018-05)
    To better understand factors influencing infectious agent dispersal within a livestock population information is needed on the nature and frequency of contacts between farm enterprises. This study uses social network analysis to describe the contact network within a vertically integrated broiler poultry enterprise to identify the potential horizontal and vertical transmission pathways for Salmonella spp. Nodes (farms, sheds, production facilities) were identified and the daily movement of commodities (eggs, birds, feed, litter) and people between nodes were extracted from routinely kept farm records. Three time periods were examined in detail, 1- and 8- and 17-weeks of the production cycle and contact networks were described for all movements, and by commodity and production type. All nodes were linked by at least one movement during the study period but network density was low indicating that all potential pathways between nodes did not exist. Salmonella spp. transmission via vertical or horizontal pathways can only occur along directed pathways when those pathways are present. Only two locations (breeder or feed nodes) were identified where the transmission of a single Salmonella spp. clone could theoretically percolate through the network to the broiler or processing nodes. Only the feed transmission pathway directly connected all parts of the network.
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    Spatial Distribution of Salmonella enterica in Poultry Shed Environments Observed by Intensive Longitudinal Environmental Sampling
    Crabb, HK ; Allen, JL ; Devlin, JM ; Wilks, CR ; Gilkerson, JR ; Elkins, CA (American Society for Microbiology, 2019-07-01)
    Detection of salmonellae within poultry environments is an important component of many food safety programs, but sampling approaches vary greatly and may not enable the detection of salmonellae when bacteria are present at a low prevalence or concentration. Intensive longitudinal sampling within caged sheds enabled us to undertake a longitudinal analysis of the spatial distribution of salmonellae in caged shed environments. Both the number of samples collected and location of sample collection within a poultry shed were important to ensure the best chance of detecting Salmonella spp. Differences in the within-shed spatial distribution of Salmonella enterica subspecies enterica serovar Typhimurium [χ2(27, 1,538) = 54.4; P < 0.001] and Salmonella enterica subspecies enterica serovar Infantis [χ2(27, 1,538) = 79.8; P < 0.0001] were identified. More than one Salmonella enterica serovar was detected in each shed on the same sampling occasion; 5% of all samples contained more than one serovar. Samples collected on the north side of the shed (odds ratio [OR], 1.77; 95% confidence interval [CI], 1.17-2.68), on the sheltered side of the shed (OR, 1.90; 95% CI, 1.26-2.89), and during winter (OR, 48.41; 95% CI, 23.56-104.19) were more likely to be positive for salmonellae. The within-shed differences observed in the both the sample prevalence and spatial location of the serovar detected indicate that there are important shed microenvironmental factors that influence the survival and/or distribution of salmonellae. These factors should be taken into consideration when environmental surveillance is undertaken for salmonellae in flocks housed in cage sheds.IMPORTANCE Routine epidemiological surveillance for salmonellae in poultry relies initially on environmental sampling. Intensive, spatially homogenous sampling, as conducted within this study, confirmed that the sampling methodology conducted within a poultry environment is a nontrivial part of sampling design. The frequency of sampling is especially important when the prevalence of Salmonella spp. is low. These factors must be taken into consideration in the design of studies for the detection of salmonellae in poultry sheds.
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    Traditional Salmonella Typhimurium typing tools (phage typing and MLVA) are sufficient to resolve well-defined outbreak events only
    Crabb, HK ; Allen, JL ; Devlin, JM ; Firestone, SM ; Stevenson, M ; Wilks, CR ; Gilkerson, JR (ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD, 2019-12-01)
    Between 1991 and 2014 the per capita notification rate of salmonellosis in Australia increased from 31.9 to 69.7 cases per 100,000 people. Salmonella Typhimurium accounted for nearly half the human cases until the end of 2014. In this study, we used cluster analysis tools to compare S. Typhimurium isolates from a chicken-meat study with those reported to the National Enteric Pathogen Surveillance System (NEPSS) from the coincident human and non-human populations. There was limited phage type diversity within all populations and a lack of specificity of MLVA profiling within phage types. The chicken-meat study isolates were not significantly clustered with the human cases and at least 7 non-human sources, based on typing profiles (PT/MLVA combination), could be implicated as a source of human cases during the same period. In the absence of a strong surveillance system representative of all putative sources, MLVA and phage typing alone or in combination are insufficient to identify the source of human cases.
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    Antimicrobials used for surgical prophylaxis by equine veterinary practitioners in Australia
    Hardefeldt, LY ; Browning, GF ; Thursky, K ; Gilkerson, JR ; Billman-Jacobe, H ; Stevenson, MA ; Bailey, KE (WILEY, 2018-01)
    BACKGROUND: Antimicrobials are widely used in Australian veterinary practices, but no investigation into the classes of antimicrobials used, or the appropriateness of use in horses, has been conducted. OBJECTIVES: The aim of the study was to describe antimicrobial use for surgical prophylaxis in equine practice in Australia. STUDY DESIGN: Cross-sectional questionnaire survey. METHODS: An online questionnaire was used to document antimicrobial usage patterns. Information solicited in the questionnaire included demographic details of the respondents, the frequency with which antimicrobials were used for specific surgical conditions (including the dose, timing and duration of therapy) and practice antimicrobial use policies and sources of information about antimicrobials and their uses. RESULTS: A total of 337 members of the Australian veterinary profession completed the survey. Generally, the choice of antimicrobial was appropriate for the specified equine surgical condition, but the dose and duration of therapy varied greatly. While there was poor optimal compliance with British Equine Veterinary Association guidelines in all scenarios (range 1-15%), except removal of a nonulcerated dermal mass (42%), suboptimal compliance (compliant antimicrobial drug selection but inappropriate timing, dose or duration of therapy) was moderate for all scenarios (range 48-68%), except for an uninfected contaminated wound over the thorax, where both optimal and suboptimal compliance was very poor (1%). Veterinarians practicing at a university hospital had higher odds of compliance than general practice veterinarians (Odds ratio 3.2, 95% CI, 1.1-8.9, P = 0.03). MAIN LIMITATIONS: Many survey responses were collected at conferences which may introduce selection bias, as veterinarians attending conferences may be more likely to have been exposed to contemporary antimicrobial prescribing recommendations. CONCLUSIONS: Antimicrobial use guidelines need to be developed and promoted to improve the responsible use of antimicrobials in equine practice in Australia. An emphasis should be placed on antimicrobial therapy for wounds and appropriate dosing for procaine penicillin.
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    Survey of veterinary prescribing for poultry disease
    Crabb, HK ; Hardefeldt, LY ; Bailey, KE ; Billman-Jacobe, H ; Gilkerson, JR ; Browning, GF (WILEY, 2019-08)
    BACKGROUND: To enable better antimicrobial stewardship, it is important to know what the patterns of prescribing behaviour are and what diseases or reasons antibiotics are being prescribed. A prescribing guideline (i.e. recommended best practice for writing prescriptions) developed by the Australian poultry veterinarians exists. However, it is not a prescribing guideline detailing treatments for the commonly observed bacterial diseases in commercial poultry. METHODS: An online survey was deployed to all registered veterinarians and members of the Australian Veterinary Poultry Association to identify prescribing behaviours for the most frequently observed bacterial diseases of poultry. RESULTS: A total of 39 survey responses were received. Most surveys were started but not completed; 13 (33%) were completed with 18 (46%) containing some information on prescribing. The most frequent treatment responses were for Escherichia coli in both layers and broilers, chronic respiratory disease (CRD), fowl cholera and spotty liver in layers and necrotic enteritis in broilers. Treatments described were for products registered for poultry use, within the recommended label dose and duration of treatment (Tables 1, 2). Unsurprisingly, tetracyclines and amoxycillin, followed by lincomycin and trimethoprim sulfonamide products were the most frequently reported treatment options. Inappropriate treatments were reported for salmonellosis and one veterinarian recommended the use of enrofloxacin for the treatment of fowl cholera. CONCLUSION: Information provided by respondents will enable the initial development of prescribing guidelines for both commercial and small poultry flocks. Importantly, it identified less than optimal prescribing behaviour for some diseases, a reliance on one class of antibiotic more than others and a failure to utilise all antimicrobial classes potentially available for treatment. Critically, the survey identified a lack of treatment options for bacterial disease in poultry. The most important bacterial diseases of poultry remain the same; effective alternatives for antibiotic treatment are required and old diseases, thought once gone, are reinventing themselves as problems for the future. Surveys of prescribing behaviours are essential for identifying diseases of high priority, changes in treatments and response to treatment and to identify areas for targeted antimicrobial stewardship, and research needs.
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    Appraisal of the Australian Veterinary Prescribing Guidelines for antimicrobial prophylaxis for surgery in dogs and cats
    Hardefeldt, LY ; Crabb, HK ; Bailey, KE ; Johnstone, T ; Gilkersoe, JR ; Billman-Jacobe, H ; Browning, GF (Wiley, 2019-07-08)
    The Australian Veterinary Prescribing Guidelines for antimicrobial prophylaxis for surgery on dogs and cats are evidence-based guidelines for veterinary practitioners. Validation of these guidelines is necessary to ensure quality and implementability. Two validated tools, used for medical guideline appraisal, were chosen to assess the guidelines. The terminology from the GuideLine Implementability Appraisal (GLIA) and the Appraisal of Guidelines for Research and Evaluation version 2 (AGREE II) were adapted for use by veterinarians. A two-phase evaluation approach was conducted. In the first phase of the evaluation, the GLIA tool was used by two specialist veterinary surgeons in clinical practice. The results of this phase were then used to modify the guidelines. In the second phase, the AGREE II tool was used by 6 general practitioners and 6 specialists to appraise the guidelines. In phase 1, the specialist surgeons either agreed or strongly agreed that the guidelines were executable, decidable, valid and novel, and that the guidelines would fit within the process of care. The surgeons were neutral on flexibility and measurability. Additional clarity around one common surgical procedure was added to the guidelines, after which the surgeons agreed that the guidelines were sufficiently flexible. In phase 2, 12 veterinarians completed the assessment using the AGREE II tool. In all sections the scaled domain score was greater than 70%. The overall quality of the guidelines was given a global scaled score of 76%. This assessment has demonstrated that the guidelines for antimicrobial prophylaxis for companion animal surgery are valid and appear implementable.
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    Serological responses of Australian horses using a commercial duplex indirect ELISA following vaccination against strangles
    El-Hage, CM ; Bannai, H ; Wiethoelter, AK ; Firestone, SM ; Heislers, CM ; Allen, JL ; Waller, AS ; Gilkerson, JR (WILEY, 2019-07-01)
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    Antimicrobial dosing for common equine drugs: a content review and practical advice for veterinarians in Australia
    Hardefeldt, LY ; Crabb, HK ; Bailey, KE ; Gilkerson, JR ; Billman-Jacobe, H ; Browning, GF (WILEY, 2019-04)
    BACKGROUND: Appropriate dosing with antimicrobial agents is critical for effective treatment and to prevent the development of antimicrobial resistance. METHODS: A review was undertaken of equine journal articles (Equine Veterinary Journal, Equine Veterinary Education, Australian Veterinary Journal, Australian Equine Veterinarian, Journal of Veterinary Internal Medicine and Journal of Equine Veterinary Science) between January 2015 and August 2018. Those with dosing regimens for procaine penicillin G, gentamicin or trimethoprim-sulfonamide in adult horses were examined and evaluated. Pharmacokinetics and -dynamics of these drugs were also reviewed. RESULTS & CONCLUSION: The most frequently reported doses for penicillin, gentamicin and trimethoprim-sulfonamide were 20-25,000 IU/kg, 6.6 mg/kg and 30 mg/kg, respectively. Veterinarians treating equine patients in Australia should be aware of the current recommended doses and inter-dosing intervals to ensure efficacy in therapy and to preserve the usefulness of these antimicrobials for the future.