Veterinary Science - Research Publications

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    NetB, a new toxin that is associated with avian necrotic enteritis caused by Clostridium perfringens
    Keyburn, AL ; Boyce, JD ; Vaz, P ; Bannam, TL ; Ford, ME ; Parker, D ; Di Rubbo, A ; Rood, JI ; Moore, RJ ; Bradley, K (PUBLIC LIBRARY SCIENCE, 2008-02-01)
    For over 30 years a phospholipase C enzyme called alpha-toxin was thought to be the key virulence factor in necrotic enteritis caused by Clostridium perfringens. However, using a gene knockout mutant we have recently shown that alpha-toxin is not essential for pathogenesis. We have now discovered a key virulence determinant. A novel toxin (NetB) was identified in a C. perfringens strain isolated from a chicken suffering from necrotic enteritis (NE). The toxin displayed limited amino acid sequence similarity to several pore forming toxins including beta-toxin from C. perfringens (38% identity) and alpha-toxin from Staphylococcus aureus (31% identity). NetB was only identified in C. perfringens type A strains isolated from chickens suffering NE. Both purified native NetB and recombinant NetB displayed cytotoxic activity against the chicken leghorn male hepatoma cell line LMH; inducing cell rounding and lysis. To determine the role of NetB in NE a netB mutant of a virulent C. perfringens chicken isolate was constructed by homologous recombination, and its virulence assessed in a chicken disease model. The netB mutant was unable to cause disease whereas the wild-type parent strain and the netB mutant complemented with a wild-type netB gene caused significant levels of NE. These data show unequivocally that in this isolate a functional NetB toxin is critical for the ability of C. perfringens to cause NE in chickens. This novel toxin is the first definitive virulence factor to be identified in avian C. perfringens strains capable of causing NE. Furthermore, the netB mutant is the first rationally attenuated strain obtained in an NE-causing isolate of C. perfringens; as such it has considerable vaccine potential.
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    A survey of veterinary consultations in Perth
    WARNE, LN ; Robertson, ID (Australian Small Animal Veterinary Association, 2003)
    Data was collected from 22 private urban veterinary practices within a 25km radius of the Perth central Post Office on the nature of 40 prospective consecutive consultations (880). Dogs and cats were presented in 96% of these consultations (70% and 26% respectively), as well as birds (3%) and other species (1%). Dogs and cats were presented more frequently for medical problems (56% and 47%) and for vaccination (32% and 35%). Surgery was performed on 17% of dogs and 20% of cats. The most frequently occurring medical problems for dogs involved the skin/coat (13%) and musculo-skeletal system (11%). For cats the most common medical problems were fight wounds and abscesses (18%) and gastrointestinal problems (6%). Neutering (dog-5%, cat-9%) and dentistry (dog-4%, cat-5%) were the most common surgical procedures performed. The results of this survey show that more dogs are taken to veterinarians in Perth than are cats, and routine health procedures, such as vaccinations and neutering, represent a significant proportion of consultations.
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    Amino acid patterns within short consensus repeats define conserved duplicons shared by genes of the RCA complex.
    McLure, CA ; Dawkins, RL ; Williamson, JF ; Davies, RA ; Berry, J ; Natalie, L-J ; Laird, R ; Gaudieri, S (Springer Science and Business Media LLC, 2004-08)
    Complement control proteins (CCPs) contain repeated protein domains, short consensus repeats (SCRs), which must be relevant to diverse functions such as complement activation, coagulation, viral binding, fetal implantation, and self-nonself recognition. Although SCRs share some discontinuous and imperfect motifs, there are many variable positions and indels making classification in subfamilies extremely difficult. Using domain-by-domain phylogenetic analysis, we have found that most domains can be classified into only 11 subfamilies, designated a, b, c, d, e, f, g, h, i, j, or k and identified by critical residues. Each particular CCP is characterized by the order of representatives of the subfamilies. Human complement receptor 1 (CR1) has ajefbkd repeated four times and followed by ch. The classification crosses CCPs and indicates that a particular CCP is a function of the mix of SCRs. The aje set is a feature of several CCPs including human CR1 and DAF and murine Crry and appears to be associated with the success or failure of implantation inter alia. This approach facilitates genomic analysis of available sequences and suggests a framework for the evolution of CCPs. Units of duplication range from single SCRs, to septamers such as efbkdaj, to extensive segments such as MCP-CR1L. Imperfections of duplication with subsequent deletion have contributed to diversification.
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    Rapid detection and non-subjective characterisation of infectious bronchitis virus isolates using high-resolution melt curve analysis and a mathematical model
    Hewson, K ; Noormohammadi, AH ; Devlin, JM ; Mardani, K ; Ignjatovic, J (SPRINGER WIEN, 2009-04-01)
    Infectious bronchitis virus (IBV) is a coronavirus that causes upper respiratory, renal and/or reproductive diseases with high morbidity in poultry. Classification of IBV is important for implementation of vaccination strategies to control the disease in commercial poultry. Currently, the lengthy process of sequence analysis of the IBV S1 gene is considered the gold standard for IBV strain identification, with a high nucleotide identity (e.g. > or =95%) indicating related strains. However, this gene has a high propensity to mutate and/or undergo recombination, and alone it may not be reliable for strain identification. A real-time polymerase chain reaction (RT-PCR) combined with high-resolution melt (HRM) curve analysis was developed based on the 3'UTR of IBV for rapid detection and classification of IBV from commercial poultry. HRM curves generated from 230 to 435-bp PCR products of several IBV strains were subjected to further analysis using a mathematical model also developed during this study. It was shown that a combination of HRM curve analysis and the mathematical model could reliably group 189 out of 190 comparisons of pairs of IBV strains in accordance with their 3'UTR and S1 gene identities. The newly developed RT-PCR/HRM curve analysis model could detect and rapidly identify novel and vaccine-related IBV strains, as confirmed by S1 gene and 3'UTR nucleotide sequences. This model is a rapid, reliable, accurate and non-subjective system for detection of IBVs in poultry flocks.
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    Behavioral aspects of electronic bull separation and mate allocation in multiple-sire mating paddocks
    Lee, C ; Prayaga, KC ; Fisher, AD ; Henshall, JM (AMER SOC ANIMAL SCIENCE, 2008-07-01)
    Controlling spatial positioning of cattle through use of electronic collars could provide new ways to farm under extensive conditions. This study examined the potential for bulls to be controlled during mating using mild electric shocks delivered through radio-controlled collars. Eighteen Belmont Red bulls were fitted with collars containing the Global Positioning System and that were able to emit a mild electric shock (500 mW) at the top of the neck behind the poll. Eighteen Belmont Red cows were fitted with Global Positioning System collars only. The experiment was replicated 3 times in 3 paddocks. Each paddock contained 2 bulls and 1 cow in induced estrus. On d 1, the bulls were either assigned to the cow or not assigned to the cow, and on d 2, the assignments were reversed, and bulls received the other treatment using a new cow. Treatments were applied for 2 h on each day. The nonassigned bull received a mild electric shock on approach to either the cow or to a bull, whereas the assigned bull received a mild electric shock on approach to the other bull only. The electric shock was applied when the bulls were within approximately 10 m and moving toward the nonallowed animal. The electric shock was terminated when the animal responded by stopping movement toward the nonallowed animal. In the first 10 min, nonassigned bulls spent less time within 5 m of the cow (P = 0.03) than assigned bulls. Assigned bulls spent more time close to the cow during the entire 120 min on d 1 than on d 2 (P = 0.014). On d 1, the assigned bulls moved more toward the cow and the nonassigned bull than they did on d 2 (P = 0.02). Assigned bulls displayed more sexual behaviors than nonassigned bulls (P = 0.004). Nonassigned bulls were sometimes observed not to approach the cow despite a change in its location. This suggests that the bull associated the electric shock with the cow and not with the location in which it received the electric shock. Instances were observed in which the cow pursued the nonassigned bull, in which case the bull did not receive an electric shock, and this may reflect the preference of the cow. This study demonstrated that bulls can be separated and prevented from approaching a cow in estrus using a mild electric shock. However, mate allocation was not completely successful due to the potential for cow preference for certain bulls.
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    A sheep cannulation model for evaluation of nasal vaccine delivery
    Yen, HH ; Scheerlinck, JPY ; Gekas, S ; Sutton, P (ACADEMIC PRESS INC ELSEVIER SCIENCE, 2006-02-01)
    We have developed and validated a novel model to investigate the efficacy of nasal vaccine delivery. Based on lymphatic cannulation of the tracheal lymph trunk of sheep, the model allows collection of lymph draining from the Nasal Associated Lymphoid Tissue. The model is suitable for determining both the amount of material that is absorbed into the lymphatic system, following intra-nasal delivery and the immune response that occurs following vaccination into the nasal area. The cell populations that track in this duct were phenotyped and found to be similar to those previously reported to be present in efferent lymph draining from peripheral lymph nodes. Following intra-nasal spray, we demonstrated that the amount of material recovered in draining lymph is only a very small fraction of the total delivered. Nevertheless, intra-nasal spraying of a vaccine could activate local immune cells. The method described will be invaluable for optimising intra-nasal delivery systems by allowing a separate optimisation of vaccine uptake and immune responses induction.
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    Protective immunity against Helicobacter is characterized by a unique transcriptional signature
    Mueller, A ; O'Rourke, J ; Chu, P ; Kim, CC ; Sutton, P ; Lee, A ; Falkow, S (NATL ACAD SCIENCES, 2003-10-14)
    Immunization with a whole-cell sonicate vaccine of Helicobacter felis in conjunction with cholera toxin as a mucosal adjuvant induces long-term protective immunity in a majority of laboratory mice. We have combined gene expression profiling and immunohistochemical analysis on a set of immunized animals to better understand the mechanism of protection. The stomachs of protected animals exhibited a strikingly different transcriptional profile compared with those of nonprotected or control mice, indicating that vaccination targets the appropriate site and leaves a molecular signature. Among the genes whose up-regulation is significantly correlated with protection are a number of adipocyte-specific factors. These include the fat-cell-specific cytokines adipsin, resistin, and adiponectin and the adipocyte surface marker CD36. Interestingly, potentially protective T and B lymphocytes can be found embedded in the adipose tissue surrounding protected stomachs but never in control or unprotected stomachs. Adipsin-specific immunohistochemical staining of protected stomach sections further revealed molecular cross-talk between adjacent lymphoid and adipose cell populations. We propose a mechanism of protection that involves the effector responses of either or both lymphocyte subclasses as well as the previously unappreciated paracrine functions of adipose tissue surrounding the resident lymphocytes.
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    Post-immunisation gastritis and Helicobacter infection in the mouse: a long term study
    Sutton, P ; Danon, SJ ; Walker, M ; Thompson, LJ ; Wilson, J ; Kosaka, T ; Lee, A (BRITISH MED JOURNAL PUBL GROUP, 2001-10-01)
    BACKGROUND AND AIMS: Helicobacter pylori is a major cause of peptic ulcers and gastric cancer. Vaccine development is progressing but there is concern that immunisation may exacerbate Helicobacter induced gastritis: prophylactic immunisation followed by challenge with H felis or H pylori can induce a more severe gastritis in mice than seen with infection alone. The aim of this study was to investigate the relationship between immunity to Helicobacter infection and post-immunisation gastritis. METHODS: (1) C57BL/6 mice were prophylactically immunised before challenge with either H felis or H pylori. Histopathology and colonisation were assessed one month post-challenge. (2) C57BL/6 mice were prophylactically immunised against H felis infection and gastritis assessed up to 18 months post-challenge. RESULTS: Prophylactic immunisation induced a reduction in bacterial colonisation following H felis challenge which was associated with increased severity of active gastritis with neutrophil infiltration and atrophy. However, immunised mice challenged with H pylori SS1 had little evidence of pathology. Long term follow up showed that post-immunisation gastritis was evident at three months. However, from six months onwards, although immunised/challenged mice still developed gastritis, there was no significant difference between inflammation in these mice and infected controls. Post-immunisation gastritis was not associated with the serum antibody response. Immunisation prevented the formation of secondary lymphoid aggregates in the gastric tissue. CONCLUSION: The H felis mouse model of post-immunisation gastritis is the most extreme example of this type of pathology. We have shown in this model that post-immunisation gastritis is a transient event which does not produce long term exacerbation of pathology.
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    Helicobacter-induced expression of Bcl-X-L in B lymphocytes in the mouse model: A possible step in the development of gastric mucosa-associated lymphoid tissue (MALT) lymphoma
    Morgner, A ; Sutton, P ; O'Rourke, JL ; Enno, A ; Dixon, MF ; Lee, A (WILEY-LISS, 2001-06-01)
    Primary gastric mucosa-associated lymphoid tissue (MALT) lymphoma may develop from chronic infection with Helicobacter sp. in the mouse model. The mechanisms of pathogenesis remain unclear. Regulation of B-cell proliferation and death are important features to investigate. Proteins encoded by bcl-2 family genes, e.g., Bcl-X(L), regulate apoptosis; and alterations in the expression of these genes can contribute to the development of cancer. Our aim was to determine the role of Bcl-X(L) in B lymphocytes in the development of gastric MALT lymphoma associated with Helicobacter infection using the BALB/c mouse model. We analyzed 37 animals with Helicobacter-associated MALT (n = 25), low-grade MALT lymphoma (n = 10) and high-grade lymphoma (n = 2), investigating the in vivo distribution of Bcl-X(L) in B cells/B-lymphoma cells using immunohistochemical analysis. In vitro cultivation of B cells/B-lymphoma cells was employed to perform RT-PCR analysis of Bcl-X(L) mRNA expression after cell stimulation with Helicobacter antigen. We found significant Bcl-X(L) protein expression in B lymphocytes within MALT and low-grade MALT lymphoma, whereas there was no and minimal expression, respectively, of Bcl-X(L) in the 2 high-grade MALT lymphoma cases. Expression of bcl-X(L) mRNA in B lymphocytes was up-regulated in vitro upon Helicobacter-antigen stimulation and associated with prolonged cell survival. These findings support the hypothesis that Bcl-X(L) plays a role in the pathogenesis of B-cell MALT lymphoma by providing cell-survival signals and by triggering the acquisition of MALT.