Evaluation of result variability with a commercial Johne's disease enzyme-linked immunosorbent assay kit and repeat testing of samples.

Two hundred one serum samples from individual dairy cows with a range of results on initial testing with a commercial Johne's disease enzyme-linked immunosorbent assay (ELISA) kit were repeat tested 5 times in each of 2 laboratories with kits produced by the same manufacturer. The results for the samples with all 10 replicates showed that the values for individual samples often had a coefficient of variation greater than 20%. As expected, the standard deviation for the results increased as the average value increased and the coefficient of variation was greater in samples with low mean values. The different lots of the commercial ELISA kit used in this study had a significant effect on both the optical density and the calculated sample to positive (S/P) ratio for test replicates. Based on the variability detected in S/P ratios of replicate samples, application of a single cutoff point to interpret individual test results as positive or negative for antibodies to Mycobacterium avium subspecies paratuberculosis could result in inconsistent classification of animals as positive or negative for Johne's disease. Such inconsistency in test interpretation leads to frustration in large animal veterinarians or producers trying to make management decisions based on individual test results. Instead of dichotomizing the test results as positive or negative based on a single cutoff value, reporting numerical values and supplying a classification scheme that includes a suspect category reflecting the uncertainty inherent in the test is recommended to provide more reliable result interpretation.     

Sortase anchored proteins of Streptococcus uberis play major roles in the pathogenesis of bovine mastitis in dairy cattle

Streptococcus uberis, strain 0140J, contains a single copy sortase A (srtA), encoding a transamidase capable of covalently anchoring specific proteins to peptidoglycan. Unlike the wild-type, an isogenic mutant carrying an inactivating ISS1 insertion within srtA was only able to infect the bovine mammary gland in a transient fashion. For the first 24 h post challenge, the srtA mutant colonised at a similar rate and number to the wild type strain, but unlike the wild type did not subsequently colonise in higher numbers. Similar levels of host cell infiltration were detected in response to infection with both strains, but only in those mammary quarters infected with the wild type strain were clinical signs of disease evident. Mutants that failed to express individual sortase substrate proteins (sub0135, sub0145, sub0207, sub0241, sub0826, sub0888, sub1095, sub1154, sub1370, and sub1730) were isolated and their virulence determined in the same challenge model. This revealed that mutants lacking sub0145, sub1095 and sub1154 were attenuated in cattle. These data demonstrate that a number of sortase anchored proteins each play a distinct, non-redundant and important role in pathogenesis of S. uberis infection within the lactating bovine mammary gland.     

Osteoarthritis in cats

Osteoarthritis is a common radiographic finding in older cats, with a prevalence of up to 90% in appendicular joints. Many cats experience impaired mobility from osteoarthritis; however, there are more and more treatment options available, including diet modification, environmental modification, medical therapy, and physical rehabilitation. Continuing challenges involve accurate diagnosis and outcome assessment of treatment, but considerable progress has been made in the last decade.     

Activin A: from sometime reproductive factor to genuine cytokine

The growth factor, activin A, was initially characterized as a putative reproductive hormone but is now known to have many other divergent roles. One of these is during inflammation. Following intravenous injection of bacterial lipopolysaccharide (LPS) into sheep, activin A is released extremely rapidly into the circulation. The release of activin A appears to be independent of fever, prostaglandins or other key proinflammatory cytokines such as TNF-alpha or IL-1beta. While the precise roles and function of this factor in inflammation are yet to be elucidated, the activin response occurs in other mammalian species besides the sheep and elevated activin has been documented for a number of clinical inflammatory conditions. Activin A therefore seems to be part of the regulatory component of the innate immune response.     

Relative sensitivity of polymerase chain reaction assays used for detection of feline herpesvirus type 1 DNA in clinical samples and commercial vaccines

OBJECTIVE: To determine relative detection rates and detection limits for 6 published polymerase chain reaction (PCR) assays used for detection of feline herpesvirus type 1 (FHV-1) DNA. SAMPLE POPULATION: 5 vaccines licensed for use in preventing FHV-1-associated disease; 15 conjunctival biopsy specimens collected from cats with keratitis, conjunctivitis, or both; and a plaque-purified field isolate of FHV-1 cultured in vitro. PROCEDURE: Vaccines and clinical samples were assessed for FHV-1 DNA by use of all 6 assays. Detection rates were calculated by assuming that any sample in which FHV-1 DNA was detected was a true-positive result. Detection limits were estimated by use of serial dilutions of DNA extracted from cultured FHV-1 and 1 clinical sample. RESULTS: Testing by use of all 6 assays resulted in detection of FHV-1 DNA in all 5 vaccines. Testing by use of all 6 assays yielded concordant results for 9 of 15 conjunctival biopsy specimens (8 with negative results and 1 with a positive result). Calculated detection rates for clinical samples ranged from 29% to 86%. Assay sensitivity was ranked similarly by use of detection rate or detection limit. CONCLUSIONS AND CLINICAL RELEVANCE: Testing by use of all assays was equally likely to detect vaccine virus. Therefore, a positive PCR result in a cat may reflect vaccine virus rather than wild-type virus. Test sensitivity as assessed by detection limits and detection rates varied greatly. Because FHV-1 can be shed in clinically normal animals, high detection rate will not necessarily correlate with high diagnostic sensitivity.