A field trial to evaluate the efficacy of a combined rotavirus-coronavirus/Escherichia coli vaccine in dairy cattle.

A field trial was designed to determine the efficacy of a combination rotavirus-coronavirus/Escherichia coli vaccine on dairy farms in southwestern Ontario. In Part A of the trial, 321 cows on 15 farms were randomly assigned to either vaccination or placebo groups. On eight farms, 50% of the dams were vaccinated, while on the other seven farms, 80% of the dams were vaccinated. In Part B of the trial, 26 farms were randomly assigned to either a total vaccination program or to no vaccination program. Mortality, disease occurrence and weight gains were recorded on all calves for the first two weeks of life. In Part A, 23.5% of all calves were treated in the first two weeks of life, 20.9% were treated specifically for scours and 3.6% of live-born calves died. Enteropathogenic E. coli was identified on 13 of the 15 farms, rotavirus on 11 and coronavirus on ten. At least one of the three potential pathogens was found on every farm. There were no significant differences between calves from placebo-treated and vaccine-treated dams with regard to the proportion treated for all diseases, or for scours, or the proportion which died. Neither were there differences in days to first treatment for all diseases (seven days on average), days to first scour (6.7 days), duration of treatments (3.9 days for all diseases, 3.7 days for scours), or estimated weight gains (0.5 kg/day to 14 days). These results were not altered when the presence or absence of enteropathogenic E. coli, rotavirus or coronavirus on the premises was accounted for.(ABSTRACT TRUNCATED AT 250 WORDS)     

Application of high-resolution melt curve analysis for classification of infectious bronchitis viruses in field specimens

Objective A real-time polymerase chain reaction (PCR)/high-resolution melt (HRM) curve analysis protocol was developed in our laboratory to differentiate infectious bronchitis (IB) virus reference strains. In the current study, this method was used to detect and classify IB viruses in field submissions. Procedure Over an 11-month period samples from 40 cases of suspected IB virus were received and 17 submissions were positive for IB virus by polymerase chain reaction. HRM curve analysis classified each strain as subgroup 1, 2 or 3 strain (12 submissions) or a strain that was unable to be classified (5 submissions). The 3′ untranslated region (UTR) and partial S1 gene nucleotide sequences for the 17 IB virus strains were determined and their identity with those of the relative reference strains compared to confirm the classifications generated using the HRM curve analysis. Results Of the 12 IB field viruses classified as subgroup 1, 2, or 3 using HRM curve analysis, the 3′UTR and S1 gene nucleotide sequences had identities ≥99% with the respective subgroup reference strain. Analysis of the 3′ UTR and S1 gene nucleotide sequences for the five IB virus strains that could not be classified indicated that four belonged to one of the subgroups, and one was a potential recombinant strain (between strains from subgroups 2 and 3). A novel recombinant strain was also detected. Conclusion HRM curve analysis can rapidly assign the majority of IB viruses present in field submissions to known subgroups. Importantly, HRM curve analysis also identified variant genotypes that require further investigation.     

Assessing the efficacy of a ram-genotyping programme to reduce susceptibility to scrapie in Great Britain

Susceptibility to clinical scrapie is associated with polymorphisms in the PrP gene. The 'ARR' allele of this gene reduces susceptibility to clinical disease caused by all known strains of the transmissible spongiform encephalopathy (TSE) agent. The British government proposes to use a ram-genotyping scheme to breed genetic resistance to clinical scrapie into the national sheep population. We considered how best to target limited genotyping resources to achieve the maximum rate of genotype evolution. We created a metapopulation model of the British sheep industry, which includes the major pure-breeds of sheep and the cross-breeds produced by crossing these pure-bred animals. The main criterion for assessing the efficacy of different strategies was the time taken to increase the prevalence of the ARR allele in the slaughter-lamb population. Our model predicted that the most-effective strategy would be to target genotyping to those rams used for pure-breeding (i.e. mated with the same breed of ewe). This strategy was compared to two further strategies, in which the proportion of rams genotyped in each breed depended on the prevalence of the ARR/ARR genotype in that breed. A policy in which the proportion of animals genotyped is reduced as the ARR prevalence in that breed increases is efficient. The most-effective policy was targeting the hill sector in the early years and gradually switching to genotyping more terminal-sire and longwool rams as the resistance of the hill sector increases.     

Hatchery-level predictive values for infectious pancreatic necrosis virus and Aeromonas salmonicida in Ontario, Canada

The probability and uncertainty of correctly classifying the IPNV and Aeromonas salmonicida status of fish-rearing and natural sites in Ontario were estimated through Monte Carlo simulations. Propagating several uncertain inputs showed the extent to which natural variability and our present lack of knowledge affect the probability of site misclassification. For the scenarios investigated, the site-level negative predictive values (SNPVs) were high and fairly constant. The site-level positive predictive values (SPPVs) - given a test specificity ranging between 0.999 and 1.0 - were much lower, more variable, and highly affected by cut-off point and sample size. Substantial uncertainty resides in classifying the pathogen status of test-positive sites, whereas much less uncertainty resides in classifying pathogen status of test-negative sites.     

Somatic cell counts in bovine milk: relationships to production and clinical episodes of mastitis.

The relationships between somatic cell counts, milk production and episodes of clinical mastitis were evaluated using data collected between 1979 and 1981 in 32 southern Ontario Holstein herds. Somatic cell counts were logarithmically transformed and the distribution of the resulting counts is presented. The seasonal pattern in cell counts was evaluated using a formal statistical procedure. Counts were lowest in the winter and spring and highest in the early fall but the differences amongst monthly geometric mean cell counts were small. Assuming a linear relationship between log somatic cell counts and test day milk production it was found that a unit increase in the log count resulted in a loss of 1.44 kg of milk. Regression analyses within specific log cell count ranges indicated that the previous estimate may underestimate losses at low cell counts and overestimate losses at higher cell counts. The relationships between cell counts and episodes of mild or acute clinical mastitis were evaluated by comparing counts preceding and following the clinical episodes to comparable counts in matched control cows. Mild cases of mastitis were preceded by higher cell counts than were found in control cows but the same phenomenon was not observed in acute cases of mastitis. Both mild and acute cases were followed by higher cell counts than were found in control cows.     

The effects of storage and method of fixation on somatic cell counts in bovine milk.

Fresh milk samples and potassium dichromate preserved milk samples were stored at both ambient, approximately 21 degree C, and refrigerator temperatures, 3-5 degree C, for varying lengths of time before somatic cell counts were performed on an electronic particle counter. Fresh milk samples stored at ambient temperatures became unacceptable for somatic cell counting by 16 hours while those stored in the refrigerator were acceptable for up to three days. Once dichromate had been added to the milk no difference in cell counts attributable to temperature of storage were detected and there was very little change with time up to 14 days. On the average the addition of the dichromate elevated the cell counts/mL. As well a method of rapid fixation of milk involving the addition of glutaraldehyde prior to counting was evaluated. In fresh milk samples the use of glutaraldehyde as a fixative required adjustment of the threshold setting on the cell counter in order to produce results comparable to those obtained from formalin fixed samples. With dichromate preserved milk samples, glutaraldehyde fixation generally elevated the cell counts but the results were variable.     

Associations between dairy production indices and lipoarabinomannan enzyme-immunoassay results for paratuberculosis.

Data from an epidemiological study in Ontario, involving 304 dairy herds, were used to identify associations between selected production indices and lipoarabinomannan antigen serological test results for paratuberculosis (LAM-ELISA). Analyses were conducted at both the herd and individual cow levels of organization. After analytically controlling for management and cow factors in the respective regression models, positive serological paratuberculosis status (as defined by the LAM-ELISA test), was associated with higher milk somatic cell counts at both the herd average (p less than 0.01), and individual cow levels of organization (p less than 0.0001). In contrast, LAM-ELISA test results were consistently not associated with calving intervals in either the herd average or individual cow level analyses. Associations between LAM-ELISA results and milk production were inconsistent. No associations were found at the herd level of organization, and LAM-ELISA results were not associated with a change in breed class average (BCA) for milk, between the previous and the most recent lactations of individual cattle. However, at the individual cow level, LAM-ELISA results were positively associated with higher milk production as measured by the current BCA (p less than 0.05), and individual cow average kg of milk produced per year of life since two years of age (p less than 0.0001).