Environmental sampling to assess the bioburden of Mycobacterium avium subspecies paratuberculosis in drylot pens on California dairies.

Mycobacterium avium subspecies paratuberculosis (MAP) is a bacterium that can cause substantial economic losses in infected dairy herds due to reduced milk production and increased cow-replacement costs. In order to control MAP in dairies with drylot pens, a standardized environmental sampling protocol to quantify MAP in fecal slurry was developed based on an existing protocol for freestall pens. Specifically, following a 24 h hold of the flush, a grab sample of approximately 10 ml of fecal slurry was collected every 1 m along the flush lane of the drylot pens, avoiding individual cow fecal pats. To determine the reliability and repatability of the new environmental sampling protocol for estimation of MAP bioburden at the pen level, two collectors simultaneously collected fecal slurry samples every day for 3 days from six drylot cow pens on two Central California dairies. During the study period no cow movement between pens was allowed with the exception of sick cows. The study herds had MAP seroprevalence of 5.8% and 3.2%, respectively, based on whole pen serum ELISA results. Variance components models for quantitative real-time PCR (qPCR) results showed samples collected from different pens on different dairies accounted for greater variablitiy in MAP concentration (65%), while samples collected by different collectors had the least variability (0.1%). In contrast, variability in MAP concentration in environmental samples collected on different days had 25% variability. The intraclass correlation coefficient showed high reliability (93%) of environmental sampling simultaneously by different collectors. In contrast, the reliability of environmental sampling at different days was 65%, which was similar to the reliability for sampling by different collectors on different days. Investigators can expect high reliability when employing the new environmental sampling protocol along with qPCR testing of environmental samples from drylot pens

Association between herd management practices and antimicrobial resistance in Salmonella spp. from cull dairy cattle in Central California.

BackgroundIn this study cull dairy cows from six California dairy herds were sampled seasonally over the course of a year. The objectives were to determine the prevalence of antimicrobial resistant (AMR) Salmonella spp. shed in cull cow feces, and the factors associated with fecal shedding of AMR and multidrug resistant (MDR) Salmonella.MethodsSix dairy farms located in the San Joaquin Valley of California were identified and enrolled as a convenience sample. On each dairy, and once during each of the four seasons, 10 cull cows were randomly selected for fecal sampling on the day of their removal from the herd. In addition, study personnel completed a survey based on responses of the herd manager to questions related to the previous 4 month's herd management and the specific cattle sampled. Fecal samples were submitted to the California Animal Health and Food Safety laboratory for Salmonella isolation. Antimicrobial resistance was evaluated using broth microdilution method and a gram-negative assay plate following Clinical Laboratory Standards Institute (CLSI) guidelines and breakpoint references. All statistical models were survey adjusted for number of animals on sampling day.ResultsA total of 62 Salmonella were isolated from 60 of the 239 fecal samples collected. For 12% (95% confidence interval (CI) [3-20]) of fecal samples a multidrug resistant Salmonella was isolated. The survey-weighted results for the two most common drug classes for which isolates were resistant were tetracycline (39%; 95% CI [27-51]) and ampicillin (18%; 95% CI [9-27]). An important finding was the identification of cephalosporin as the third most common drug class for which isolates were resistant, with ceftriaxone (10%; 95% CI [2-17]) being the most common drug associated with resistance in that class. At the cow-level, reason for culling, prior treatment with antimicrobial drugs as the reason for culling was associated with higher odds of isolating an AMR Salmonella isolate. At the herd-level, percent of animals monthly culled on the farm as well as number of milking cows in the herd were associated with isolation of antimicrobial resistant Salmonella in cull cows.DiscussionSalmonella isolated from fecal samples from cull cows were resistant to important antimicrobials, such as ceftriaxone. The most common drug classes for which isolates were resistant were tetracyclines and beta-lactams, with ampicillin, ceftriaxone and ceftiofur being the three most common drugs within the latter. Cow and herd level factors were associated with isolating antimicrobial resistant Salmonella that should be further investigated for their potential role in promoting occurrence of AMR Salmonella. Our results also highlight the importance of monitoring dairy cattle sent to slaughter for shedding of Salmonella resistant to medically important antimicrobial drugs

Bayesian estimation of diagnostic accuracy of fecal culture and PCR-based tests for the detection of Salmonella enterica in California cull dairy cattle.

Epidemiological studies of low prevalence disease problems are often hindered by the high cost of diagnostic testing. The objective of this study was to evaluate PCR screening of both individual and pooled fecal samples from culled dairy cows for the invA gene of Salmonella followed by culture to determine if the sensitivity and specificity were comparable to the results from traditional culture methods applied to individual samples. Cows from six different dairies were sampled in all four seasons. A total of 240 individual cow fecal samples, 24 fecal pools and 24 pools of 24-hour tetrathionate enrichment broth were tested. Diagnostic sensitivity of PCR screening followed by culture of PCR positive or indeterminate samples (i.e PCR-CUL method) was lower than that of culture (CUL) when applied to individual fecal samples (94.8%, 99.5%), however the specificity was comparable (99.6% and 97.7% respectively). For pools of five fecal samples and pools of five, 24 h tetrathionate broth samples, the specificity of both tests were comparable (∼98%); however, their sensitivity was only comparable in pooled fecal samples (∼93%) but greater for culture compared to PCR-CUL in pooled broth samples (∼99% versus ∼93%). Compared to culture results from testing of individual fecal samples, testing pooled fecal samples by culture had a relative sensitivity of 74% and relative specificity of 96%, testing pooled fecal samples by PCR-CUL resulted in relative sensitivity of 90% and relative specificity of 96%. Testing of pooled 24-hour enrichment broth by PCR-CUL increased the relative sensitivity and specificity to 100%. PCR testing followed by culture of positive or indeterminate samples is a time saving alternative to traditional methods. In addition, pooling of samples may be a useful method for decreasing cost if study aims can accommodate a moderate loss of relative sensitivity

Diagnostics in animal health: How UC helps exclude and minimize impact of livestock pathogens

UC has a wide reach in the agriculture sector of the California economy and is well recognized for research expertise in plant diseases. Less well known is the role UC plays in animal agriculture. In 2012, the California Animal Health and Food Safety lab at UC Davis performed nearly 980,000 tests on samples from sick livestock, including cattle, horses, pigs, chickens and turkeys. The lab is prepared to respond rapidly to any disease outbreak or identification of a foreign disease. Researchers at the School of Veterinary Medicine at UC Davis are testing novel subunit vaccines to prevent pinkeye in cattle; UC ANR specialists and advisors and the staff at the Sierra Foothill Research and Extension Center were key to the development of best management practices that landowners and resource managers are using to protect their herds and public water sources against the parasite Cryptosporidium parvum; and UC veterinary scientists are part of a large team of experts, including state and federal agencies, determined to combat the endemic bluetongue virus, which can affect the state's exports

Bovine cardiac mesothelial hyperplasia: a common incidental finding in adult cattle.

Cardiac mesothelial hyperplasia forming pale plaque lesions on the epicardial surface is a common incidental finding in the hearts of aged humans. A similar phenomenon with a more papillary appearance has also been reported as an incidental finding in dogs and mice. These lesions are believed to occur in response to friction between the epicardium and overlying pericardium. We investigated this lesion in adult cattle, a phenomenon that has been associated with bovine leukemia virus infection and epicardial lymphoma. We examined 73 hearts from adult cattle, predominantly of dairy breeds: 53 from a rendering facility and 20 from a state diagnostic laboratory. Cardiac mesothelial hyperplasia was much more prevalent in cattle than in other reported species (97% of examined hearts). The most common distribution was overlying the great vessels in a dark red papillary pattern. Cardiac mesothelial hyperplasia was also variably observed on all 4 cardiac chambers and the pericardium. Occasionally these lesions took on a smooth plaque-like appearance resembling those observed in humans. The lesions varied from 0.25 cm2 to covering 90% of the epicardial surface. No association was observed between cardiac mesothelial hyperplasia and bovine leukemia virus infection or cardiac lymphoma. Cardiac mesothelial hyperplasia was a common incidental finding in bovine hearts that must be distinguished from neoplasia and acute or chronic inflammation

Bovine cardiac mesothelial hyperplasia: a common incidental finding in adult cattle.

Cardiac mesothelial hyperplasia forming pale plaque lesions on the epicardial surface is a common incidental finding in the hearts of aged humans. A similar phenomenon with a more papillary appearance has also been reported as an incidental finding in dogs and mice. These lesions are believed to occur in response to friction between the epicardium and overlying pericardium. We investigated this lesion in adult cattle, a phenomenon that has been associated with bovine leukemia virus infection and epicardial lymphoma. We examined 73 hearts from adult cattle, predominantly of dairy breeds: 53 from a rendering facility and 20 from a state diagnostic laboratory. Cardiac mesothelial hyperplasia was much more prevalent in cattle than in other reported species (97% of examined hearts). The most common distribution was overlying the great vessels in a dark red papillary pattern. Cardiac mesothelial hyperplasia was also variably observed on all 4 cardiac chambers and the pericardium. Occasionally these lesions took on a smooth plaque-like appearance resembling those observed in humans. The lesions varied from 0.25 cm2 to covering 90% of the epicardial surface. No association was observed between cardiac mesothelial hyperplasia and bovine leukemia virus infection or cardiac lymphoma. Cardiac mesothelial hyperplasia was a common incidental finding in bovine hearts that must be distinguished from neoplasia and acute or chronic inflammation