Distribution and temporal prevalence of Theileria orientalis major piroplasm surface protein types in eastern Australian cattle herds.

OBJECTIVE: To determine the distribution of theilerial types in eastern Australian cattle herds and their changing prevalence in regions of New South Wales (NSW). DESIGN: Survey testing of herds in 2010-11 in Queensland (QLD), NSW and Victoria (VIC) where clinical theileriosis was not evident and ongoing surveillance in NSW through laboratory submissions. METHODS: Blood samples were tested by PCR targeting the Theileria orientalis p32 gene and positive tests were examined for the Ikeda, Chitose and Buffeli types. Survey samples from 516 cattle in 50 herds and diagnostic submissions from 434 suspect field cases in 116 herds were analysed. RESULTS: In clinically normal survey cattle, T. orientalis prevalence was high (NSW 23.7%, QLD 56.8%, VIC 34.0%), with variability among regions of each state. Chitose was the most common and widespread type (19.1-43.7% per state), with Buffeli present in all states at a lower prevalence (10.8-24.8% per state). Ikeda was detected in three of five regions in QLD (North, South and South East; prevalence 3.4-15.4%), only one of the surveyed regions in NSW (North Coast; prevalence 74.2%) and in only one animal in VIC. Evidence of clinical disease and laboratory confirmation of Ikeda infection in diagnostic submissions were predominant in several NSW regions, with increasing numbers of affected herds particularly in the coastal Mid-Coast and Cumberland areas. CONCLUSIONS: In those regions where prior evidence of theileriosis was uncommon, Ikeda infection was evident in a limited number of NSW regions and multiple regions in QLD. However, clinical disease has continued to become widespread in NSW and VIC, involving Ikeda strains in many regions.

Evaluation of serovar-independent ELISA antigens of Actinobacillus pleuropneumoniae in pigs following vaccination or experimental challenge with respiratory pathogens and natural A. pleuropneumoniae serovar 1 challenge.

OBJECTIVE: To compare the sensitivity and cross-reactivity of six serological enzyme-linked immunosorbent assays (ELISAs) based on serovar-independent antigens of Actinobacillus pleuropneumoniae (App). DESIGN: An experimental pig trial using direct or natural challenge with App and direct challenge or vaccination using other common respiratory pathogens. PROCEDURE: A 39-kDa outer membrane protein antigen and five recombinant antigens from the ApxIVA gene of App were evaluated. The latter were derived from the ApxIVA N terminus (ApxIVA-N, ApxIVA-NP, ApxIVA-NPS) or C terminus (ApxIVA-C, ApxIVA-CP). Pigs were sampled after direct challenge with App, Pasteurella multocida or Haemophilus parasuis, after vaccination with these organisms and after natural App infection. Clinical and necropsy findings were evaluated. RESULTS: The 39-kDa ELISA had high sensitivity, but cross-reactivity, following P. multocida challenge. ELISAs using ApxIVA N terminus antigens were clearly more sensitive than C terminus antigens for detection of App-induced disease. Although affinity-purified ApxIVA-NP antigen detected marginally more diseased pigs than the -N and -NPS ELISAs, these assays only detected 41-47% of 17 pigs with lung lesions and microbiological evidence of App based on sampling up to 4-5 weeks after natural (13 pigs) or 5 weeks after direct App serovar 1 challenge (4 pigs). CONCLUSIONS: The 39-kDa ELISA readily detects App exposure and infection, but is adversely affected by P. multocida infection. ApxIVA-N-based ELISAs can be used to evaluate the App status of commercial herds, but a proportion of infected and diseased animals are seronegative at 1 month after likely exposure to aerosol transmission of App from clinical cases.

Evaluation of serovar-independent ELISA antigens of Actinobacillus pleuropneumoniae in pigs, following experimental challenge with A. pleuropneumoniae, Mycoplasma hyopneumoniae and Pasteurella multocida.

OBJECTIVE: To compare the sensitivity and specificity of six serological enzyme-linked immunosorbent assays (ELISAs) based on serovar-independent antigens of Actinobacillus pleuropneumoniae (App) and investigate cross-reactivity in disease-free pigs challenged with Mycoplasma hyopneumoniae and Pasteurella multocida. DESIGN: Five experimental pig trials using direct challenge with App serovars 1, 7 or 15 or direct challenge with M. hyopneumoniae and/or various dose rates of P. multocida. PROCEDURE: A 39-kDa outer membrane protein antigen and five recombinant antigens from the apxIVA gene of App were evaluated. The latter were derived from the ApxIVA N-terminus (ApxIVA-N, ApxIVA-NP, ApxIVA-NPS) or C-terminus (ApxIVA-C, ApxIVA-CP). Pigs were sampled after challenge and clinical and necropsy findings evaluated. RESULTS: The 39-kDa ELISA had high sensitivity but lacked specificity, with significantly increased cross-reactivity following P. multocida challenge. ELISAs based on ApxIVA N-terminus antigens were significantly more sensitive than C-terminus antigens for the detection of App-induced disease. Although ApxIVA-N and ApxIVA-NP ELISAs had increased reactivity following P. multocida challenge, they retained high specificity for App-induced disease (90-93%). Affinity purified ApxIVA-NP antigen had marginally better specificity than ApxIVA-N, without reduced sensitivity. Mycoplasma hyopneumoniae did not affect serological cross-reactivity. In disease-free pigs, the specificity of the ApxIVA-NPS ELISA may be adversely affected by nasal carriage of apparently low-virulence App strains. CONCLUSIONS: ApxIVA-N-based ELISAs can be used for evaluating App status in commercial herds, but some appear limited by high carriage rates of low-virulence App. The 39-kDa antigen is only of merit in exclusion of App disease by negative serology.

Bacterial survival studies to assess the efficacy of static pile composting and above ground burial for disposal of bovine carcases.

AIM: To assess the survival of bacteria during two alternative means of cattle carcase disposal in windrows: static pile composting (SPC) and above ground burial in soil (AGB), under temperate climate conditions on agricultural land, compared to surface disposal as the control method. METHODS AND RESULTS: Bacteriological reference materials (pooled bovine faeces in permeable nylon bags and lyophilized cultures of Escherichia coli in glass ampoules) were positioned above and below each of 33 beef cattle carcases (250-300 kg). Temperatures at these sites were monitored with data loggers, while temperature and CO(2) probes were applied repeatedly at varying depths along the windrows. Aliquots of each reference material were cultured from three randomly selected animals from the SPC and AGB group and from all three control animals on five occasions (at 28, 56, 84, 126 and 182 days). SPC was highly efficacious in the destruction of coliforms in faeces and E. coli in ampoules within 28 days, while AGB was not significantly better than controls until 84 days, and bacteria in reference materials above the AGB carcases were still viable after 182 days. Temperature probes and loggers showed SPC provided sustained temperatures of 55-70°C, while AGB did not reach temperatures of 30°C, and the temperature differences correlated with bacteriological findings. CONCLUSIONS: In relation to emergency disease management, SPC can be successfully applied to eliminate pathogenic bacteria in cattle carcases, but AGB is unsuitable for carcase disposal. SIGNIFICANCE AND IMPACT OF THE STUDY: In emergency, animal disease outbreaks in temperate climates requiring large-scale ruminant carcase disposal, SPC can be successfully applied for the destruction of micro-organisms.

Serological responses to two serovar-independent ELISA antigens of Actinobacillus pleuropneumoniae in Australian commercial pig herds.

OBJECTIVE: To compare serological responses in pig herds classified as low or high risk for disease caused by Actinobacillus pleuropneumoniae, using two ELISA tests based on serovar-independent antigens. PROCEDURE: Cross-sectional sampling was undertaken in 13 commercial herds, the clinical and slaughter histories of which indicated either freedom from (n = 5) or prior confirmed cases of A. pleuropneumoniae (n = 8). In nine herds, approximately 40 pigs each were sampled at 4, 8, 12, 16 and 20 weeks. Three of the remaining four herds were sampled between 6 and 30 weeks of age, and the last was sampled only prior to slaughter, at approximately 24 weeks. Sera were tested in ELISA based on two antigens common among A. pleuropneumoniae serovars: a 39-kDa outer membrane protein and a recombinant ApxIVA-N terminus protein. RESULTS: Sampling of 1 and 5 to 6-month-old pigs provided the most useful information on herd status. The 39-kDa ELISA was sensitive in detecting infected herds, but had evidence of cross-reactivity with high seroreactivity rates in older pigs in some low-risk herds. The ApxIVA-N ELISA was less seroreactive in high-risk herds and had higher specificity in low-risk herds. CONCLUSION: ELISA based on the 39-kDa subunit are of limited use, because of possible cross-reactivity, but a high negative predictive value may be useful for risk assessment in suspect herds. Maternal antibody to ApxIVA-N may be of value in detecting high-risk herds, but 5% of 4-week-old pigs in low-risk herds were also seropositive in this assay.