Biomedical subjects

Ian A Gardner

Publications and source records attributed to Ian A Gardner.

At least 19 recordsLinked to original sources

PubMed2006

Consensus recommendations on diagnostic testing for the detection of paratuberculosis in cattle in the United States.

The report provided here contains a simplified set of diagnostic testing recommendations. These recommendations were developed on the basis of research funded by the USDA-Animal and Plant Health Inspection Service-Veterinary Services through a cooperative agreement. The report is intended to provide simple, practical, cost-effective consensus testing recommendations for cattle herds that are not enrolled in the US Test-Negative Program. The information has been reviewed by paratuberculosis (Johne's disease) experts at the USDA and academic centers as well as stakeholders in various segments of the cattle industry. The recommendations were accepted by the National Johne's Working Group and Johne's Disease Committee of the US Animal Health Association during their annual meetings in October 2006. The report is intended to aid veterinarians who work with cattle producers in the United States. The recommendations are based on information available up to October 2006. There is a paucity of large-scale, high-quality studies of multiple tests conducted on samples obtained from the same cattle. It is understood that there may be special circumstances that require deviation from these recommendations. Furthermore, as new information becomes available and assays are improved and their accuracy is critically evaluated, changes to these recommendations may be necessary.

Animals↗

PubMed2006

Efficacy of spheroplastic and cell-wall competent vaccines for Mycobacterium avium subsp. paratuberculosis in experimentally-challenged baby goats.

A Mycobacterium avium subspecies paratuberculosis (MAP) vaccine that reduced the incidence of clinical disease or reduced fecal shedding of MAP would aid control of Johne's disease (JD). The objective of the present study was to evaluate the efficacy of four MAP vaccine combinations, including cell-wall competent (CWC) alum adjuvant, CWC-QS21 adjuvant, cell-wall deficient (CWD) alum adjuvant and CWD-QS21 adjuvant vaccines. Eighty baby goats were vaccinated at 1 and 4 weeks of age with one of these vaccines or a sham control vaccine consisting of alum adjuvant. Kids were challenged orally with approximately 6.0x10(9) organisms in four divided doses of 1.5x10(9) organisms using a goat isolate of MAP. Vaccinated challenged and challenged control groups had 10 and 6 kids per group, respectively. Half of the kids within each group were necropsied at either 6 or 9 months post-challenge. Gross and microscopic lesions and relative number of acid-fast bacilli were evaluated and scored at necropsy. Results indicated all challenged kids had some lesions compatible with JD suggesting none of the vaccines prevented infection. Three vaccines (CWC-alum, CWC-QS21 and CWD-QS21) reduced lesion scores by 46-51% at 9 months. CWD-alum vaccine resulted in a more severe (+33.5%) lesion score than sham-vaccinated challenged control. Lesion scores were greater at 9 months than at 6 months post-challenge in the sham-vaccinated challenged group and CWD-alum vaccinated group, while lesion scores were generally stable with remaining vaccines. Mean fecal CFU/g were significantly different across time from challenge to 9-month necropsy (p=0.043) and the CWC-QS21 vaccine group had a marked reduction in fecal CFU/g at all time points post-challenge. A reduction in MAP CFU/g was also detected in necropsy tissues from kids given the CWC-alum, CWC-QS21 and CWD-QS21 vaccines, and increased CFU/g were detected in tissues from kids given the CWD-alum vaccine. Immunological tests evaluated included, humoral response evaluation by AGID, ELISA and Western blot, and cell mediated response by comparative PPD skin testing (M. avium, Old Johnin, M. bovis and Lot 2 Johnin PPD's), and production of MAP induced gamma-interferon. Vaccination also resulted in false-positive PPD skin test reactions for M. avium PPD, Old Johnin PPD and gamma-interferon tests. When a 2-mm cutoff above normal skin thickness was used to define positive skin test reactions, false-positive reactions for M. bovis were detected in only 2 of 32 kids given a vaccine with QS21 adjuvant.

Animals↗

PubMed2006

Recombinant canarypox virus vaccine co-expressing genes encoding the VP2 and VP5 outer capsid proteins of bluetongue virus induces high level protection in sheep.

We describe the development and preliminary characterization of a recombinant canarypox virus vectored vaccine for protective immunization of ruminants against bluetongue virus (BTV) infection. Sheep (n=6) immunized with recombinant canarypox virus vector (BTV-CP) co-expressing synthetic genes encoding the two outer capsid proteins (VP2 and VP5) of BTV serotype 17 (BTV-17) developed high titers (40-160) of virus-specific neutralizing antibodies and were resistant to challenge with a field strain of BTV-17. In contrast, sheep (n=5) immunized with a commercial recombinant canarypox virus vector expressing the E and preM genes of West Nile virus were seronegative to BTV and developed pyrexia, lymphopenia, and extended, high-titered viremias following challenge exposure to the field strain of BTV-17. These data confirm that the BTV-CP vaccine may be useful for the protective immunization of ruminants against bluetongue, and it may avoid the problems inherent to live-attenuated (LA) BTV vaccines.

Animals↗

PubMed2006

Sample size calculations for disease freedom and prevalence estimation surveys.

We developed a Bayesian approach to sample size calculations for studies designed to estimate disease prevalence that uses a hierarchical model for estimating the proportion of infected clusters (cluster-level prevalence) within a country or region. The clusters may, for instance, be villages within a region, cities within a state, or herds within a country. Our model allows for clusters with zero prevalence and for variability in prevalences among infected clusters. Moreover, uncertainty about diagnostic test accuracy and within-cluster prevalences is accounted for in the model. A predictive approach is used to address the issue of sample size selection in human and animal health surveys. We present sample size calculations for surveys designed to substantiate freedom of a region from an infectious agent (disease freedom surveys) and for surveys designed to estimate cluster-level prevalence of an endemic disease (prevalence estimation surveys). In disease freedom surveys, for instance, assuming the cluster-level prevalence for a particular infectious agent in the region is greater than a maximum acceptable threshold, a sample size combination consisting of the number of clusters sampled and number of subjects sampled per cluster can be determined for which authorities conducting the survey detect this excessive cluster-level prevalence with high predictive probability. The method is straightforward to implement using the Splus/R library emBedBUGS together with WinBUGS.

Animals↗

PubMed2006

Outdoor fecal deposition by free-roaming cats and attitudes of cat owners and nonowners toward stray pets, wildlife, and water pollution.

OBJECTIVE: To estimate cat population size, management, and outside fecal deposition and evaluate attitudes of cat owners and nonowners to stray animal control, water pollution, and wildlife protection. DESIGN: Cross-sectional survey. SAMPLE POPULATION: 294 adult residents of Cayucos, Los Osos, and Morro Bay, Calif. PROCEDURES: Telephone survey. RESULTS: The region's cat population was estimated at 7,284 owned and 2,046 feral cats, and 38% of surveyed households owned a mean of 1.9 cats/household. Forty-four percent of cats defecated outside >75% of the time. Annual fecal deposition (wet weight) by owned cats in the 3 communities was estimated to be 77.6 tonnes (76.4 tons). Cat owners were more likely to oppose cat licensing and impounding stray cats and support trap-neuter-return for stray cats and less likely to be concerned about water pollution, than were noncat owners. CONCLUSIONS AND CLINICAL RELEVANCE: Feral cats represented a sizeable proportion (22%) of the free roaming cats in this area and could be contributing 30.0 tonnes (29.5 tons) of feces to the environment per year. However, feral cats are not the principal source of fecal loading because owned cats defecating outdoors contribute an estimated 77.6 tonnes (76.4 tons) or 72% of the annual outdoor fecal deposition.

Animal Husbandry↗

PubMed2006

Temporal detection of equine herpesvirus infections of a cohort of mares and their foals.

The objectives of this study were to estimate the prevalence of equine herpesviruses (EHV) 1-5 in the nasal secretions (NS) of a cohort of 12 mares and their foals from birth to 6 months of age, estimate the prevalence of EHV-1-5 infection of peripheral blood mononuclear cells (PBMC) of selected foals, and investigate phylogenetic relationships amongst the various strains of EHV-2 and 5. Virus-specific PCR assays were used to detect EHV-1-5 in NS and PBMC. A homologous portion of the glycoprotein B (gB) gene of the various strains of EHV-2 and 5 was sequenced and compared. EHV-2, 4, and 5 were all detected in NS from the horses, but only EHV-4 was associated with respiratory disease (P=0.005). EHV-2 and 5 infections were both common, but foals shed EHV-2 in their NS earlier in life than EHV-5 (P=0.01). Latent EHV-2 and 5 infections were detected in the PBMC of 75 and 88%, respectively, of the foals at approximately 6 months of age. The strains of EHV-2 shed in the NS of individual horses were more genetically heterogeneous than the strains of EHV-5 (95.5-99.3% versus 98.8-99.3% nucleotide identity, respectively). One-month-old foals typically shed strains of EHV-2 that were identical to those infecting their dams whereas older foals often shed virus strains that were different from those of their dams. Although herpesvirus infections were ubiquitous in this cohort of horses, there were distinct clinical consequences and clear epidemiological differences between infections with the different viruses.

Aging↗

PubMed2006

Sensitivity and specificity of the agar-gel-immunodiffusion test, ELISA and the skin test for detection of paratuberculosis in United States Midwest sheep populations.

Our objective was to estimate the sensitivity and specificity of the agar-gel-immunodiffusion test (AGID), the ELISA, and the skin test for the detection of Mycobacterium avium subspecies paratuberculosis (MAP) in sheep using Bayesian methods without a gold standard. Fourteen flocks (2 465 sheep) were used. Five flocks (450 sheep) were considered MAP non-infected and 9 flocks (2 015 sheep) had sheep infected with MAP. Sheep were skin tested and blood was collected for AGID and ELISA testing. Results were analyzed using a Bayesian 3-test in 1-population model fitted in WinBUGS. The model allowed for dependence (correlation) between the two serologic tests, but these two tests were assumed to be conditionally independent of the skin test. The estimated specificity was 99.5% (95% PI of 98.9-99.9%) for the AGID; 99.3% (98.4-99.8%) for the ELISA using an optical density measured cutoff of 0.20; 99.2% (98.1-99.8%) using a cutoff of 0.15; 97.5% (95.8-98.7%) using a cutoff of 0.10; and 98.7% (97.3-99.5%) for the skin test. The estimated sensitivities were 8.3% (6.2-10.7%) for the AGID; 8.0% (6.0-10.4%), 10.6% (8.3-13.1%), and 16.3% (13.5-19.4%) for the ELISA using the cutoffs 0.20, 0.15, and 0.10 respectively; and 73.3% (62.3-85.8%) for the skin test. The skin test was specific in non-infected populations and sensitive in infected populations, although in some cases a positive skin test might represent MAP exposure rather than infection. The AGID and ELISA were specific but lacked sensitivity. The AGID and ELISA consistently identified two different populations of infected sheep with only moderate overlap between positive test results.

Animals↗

PubMed2006

Modified culture protocol for isolation of Mycobacterium avium subsp. paratuberculosis from raw milk.

A modified culture method using C18-carboxypropylbetaine (CB-18) and microscopic screening was evaluated for time to and limit of detection of Mycobacterium avium subsp. paratuberculosis (MAP) in raw milk. Bulk-tank milk samples were spiked with six different concentrations (10(1) to 10(6) CFU/mL) of MAP. Samples were processed using two different protocols. The first protocol involved specimen processing with the zwitterionic detergent C18-carboxypropylbetaine (CB-18) and lytic enzymes followed by culture on modified Middlebrook 7H10 agar plates with microscopic screening. The second protocol used 0.75% hexadecylpyridinium chloride (HPC) for specimen processing, followed by culture on Herrold's egg yolk medium (HEYM). Both protocols were repeated eight independent times, and the detection limit and time to detection were compared. The presence of MAP in spiked milk samples was detected between 14 and 45 days (N [number of samples], 46; mean, 22.7; median, 19.5) using the CB-18 and microscopic screening method, and between 21 and 63 days (N, 47; mean, 31; median, 28) using HEYM (P < 0.001). Time to detection also varied with MAP concentration (P < 0.001). Higher concentrations of MAP were detected earlier than lower concentrations and this finding was independent of the method used (P = 0.479). The two methods had similar detection limits but the modified culture method reduced the time to detect MAP in raw milk for the majority of concentrations.

Animals↗

PubMed2006

Receiver-operating characteristic curves and likelihood ratios: improvements over traditional methods for the evaluation and application of veterinary clinical pathology tests.

Receiver-operating characteristic (ROC) curves provide a cutoff-independent method for the evaluation of continuous or ordinal tests used in clinical pathology laboratories. The area under the curve is a useful overall measure of test accuracy and can be used to compare different tests (or different equipment) used by the same tester, as well as the accuracy of different diagnosticians that use the same test material. To date, ROC analysis has not been widely used in veterinary clinical pathology studies, although it should be considered a useful complement to estimates of sensitivity and specificity in test evaluation studies. In addition, calculation of likelihood ratios can potentially improve the clinical utility of such studies because likelihood ratios provide an indication of how the post-test probability changes as a function of the magnitude of the test results. For ordinal test results, likelihood ratios can be calculated on a category-specific basis from the empirical data or by using the slope of the line joining adjacent category limits on the ROC curve. For continuous test results, data need to be categorized into intervals for estimation of likelihood ratios, or they can be calculated as the slope (tangent) to the ROC curve at a unique test value. We use ROC analysis and calculate likelihood ratios to evaluate the performance of tests reported in 2 articles previously published in this journal.

Animal Diseases↗

PubMed2006

Use of age and milk production data to improve the ability of enzyme-linked immunosorbent assay test results to predict Mycobacterium avium ssp. paratuberculosis fecal culture status.

Cows from 2 California dairies were tested for paratuberculosis at the end of lactation by using fecal culture and a commercially available serum enzyme-linked immunosorbent assay (ELISA) test kit. Individual cow characteristics and production variables were evaluated along with ELISA testing results as predictors of fecal culture status. In multivariable logistic regression analysis, age and a herd-standardized version of 305-day mature equivalent (305 ME) milk production were significant predictors of fecal culture status after adjusting for herd, quarter of the study year, and ELISA sample-to-positive (S/P) ratio. The area under a nonparametric receiver operating characteristic curve was significantly greater for a multivariable model that included age and the level of milk production when compared with a model without these covariates. In conclusion, consideration of cow-level covariates was useful as an aid in predicting Mycobacterium avium ssp. paratuberculosis (MAP) fecal culture status. For a given ELISA S/P ratio, older cows and those with lower 305 ME milk production relative to other cows in the herd were significantly more likely to be shedding MAP in their feces at the end of lactation.

Age Factors↗

PubMed2006

Detection of antibodies to West Nile virus in equine sera using microsphere immunoassay.

One hundred and ninety-one sera from horses that recently were exposed to West Nile virus (WNV) by either vaccination or natural infection or that were not vaccinated and remained free of infection were used to evaluate fluorescent microsphere immunoassays (MIAs) incorporating recombinant WNV envelope protein (rE) and recombinant nonstructural proteins (rNS1, rNS3, and rNS5) for detection of equine antibodies to WNV. The rE MIA had a diagnostic sensitivity and specificity, respectively, of 99.3% and 97.4% for detection of WNV antibodies in the serum of horses that were recently vaccinated or naturally infected with WNV, as compared to the plaque reduction neutralization test (PRNT). The positive rE MIA results were assumed to be WNV-specific because of the close agreement between this assay and the PRNT and the fact that unvaccinated control horses included in this study were confirmed to be free of exposure to the related St Louis encephalitis virus. The NS protein-based MIA were all less sensitive than either the rE MIA or PRNT (sensitivity 0-48.0), although the rNSI MIA distinguished horses vaccinated with the recombinant WNV vaccine from those that were immunized with the inactivated WNV vaccine (P < 0.0001) or naturally infected with WNV (P < 0.0001). The rE MIA would appear to provide a rapid, convenient, inexpensive, and accurate test for the screening of equine sera for the presence of antibodies to WNV.

Animals↗

PubMed2006

Evaluation of cost-effectiveness of targeted sampling methods for detection of Mycobacterium avium subsp paratuberculosis infection in dairy herds.

OBJECTIVE: To investigate the epidemiologic and financial impacts of targeted sampling of subpopulations of cows, compared with random sampling of all cows, for classification of dairy herd infection status for paratuberculosis. ANIMALS: All cows from 4 infected herds with a low-to-moderate prevalence of paratuberculosis and from 1 noninfected herd in California. PROCEDURE: The infection status of each cow was classified on the basis of results of an ELISA or combined ELISA and fecal culture results. Thirteen sampling schemes designed to randomly sample cows on the basis of lactation number, stage of lactation, and milk production were evaluated. Sampling without replacement was used to obtain a probability of herd detection of paratuberculosis for each evaluated sampling method and for simulated sample sizes between 30 and 150 cows. Marginal cost-effectiveness analysis was used to determine the cost increase relative to the increase in detection probability. RESULTS: Sampling cows in the third or higher lactation and > or = 200 days into lactation yielded the highest detection probability in most instances, resulting in a detection probability that was 1.4 to 2.5 times that obtained by sampling 30 cows in the second or higher lactation. Costs of testing via the alternative method with a 95% detection probability were approximately dollar 300 lower in a high-prevalence herd (31%) and dollar 800 lower in a low-prevalence herd (9%), compared with use of the reference method. CONCLUSIONS AND CLINICAL RELEVANCE: Detection of herds with paratuberculosis could be improved, and costs of testing substantially reduced by sampling targeted groups of cows.

Animals↗

PubMed2006

Association between findings on palmarodorsal radiographic images and detection of a fracture in the proximal sesamoid bones of forelimbs obtained from cadavers of racing Thoroughbreds.

OBJECTIVE: To determine the distribution for limbs and bones in horses with fractures of the proximal sesamoid bones and relationships with findings on palmarodorsal radiographic images. SAMPLE POPULATION: Proximal sesamoid bones obtained from both forelimbs of cadavers of 328 racing Thoroughbreds. PROCEDURE: Osteophytes; large vascular channels; and fracture location, orientation, configuration, and margin distinctness were categorized by use of high-detail contact palmarodorsal radiographs. Distributions of findings were determined. Relationships between radiographic findings and fracture characteristics were examined by use of chi2 and logistic regression techniques. RESULTS: Fractures were detected in 136 (41.5%) horses. Biaxial fractures were evident in 109 (80%) horses with a fracture. Osteophytes and large vascular channels were evident in 266 (81%) and 325 (99%) horses, respectively. Medial bones typically had complete transverse or split transverse simple fractures, indistinct fracture margins, > 1 vascular channel that was > 1 mm in width, and osteophytes in abaxial wing and basilar middle or basilar abaxial locations. Lateral bones typically had an oblique fracture and distinct fracture margins. Odds of proximal sesamoid bone fracture were approximately 2 to 5 times higher in bones without radiographic evidence of osteophytes or large vascular channels, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Biaxial fractures of proximal sesamoid bones were common in cadavers of racing Thoroughbreds. Differences between medial and lateral bones for characteristics associated with fracture may relate to differences in fracture pathogeneses for these bones. Osteophytes and vascular channels were common findings; however, fractures were less likely to occur in bones with these features.

Age Distribution↗

PubMed2006

Indirect fluorescent antibody testing of cerebrospinal fluid for diagnosis of equine protozoal myeloencephalitis.

OBJECTIVE: To assess the use of CSF testing with an indirect fluorescent antibody test (IFAT) for diagnosis of equine protozoal myeloencephalitis (EPM) caused by Sarcocystis neurona. SAMPLE POPULATION: Test results of 428 serum and 355 CSF samples from 182 naturally exposed, experimentally infected, or vaccinated horses. PROCEDURE: EPM was diagnosed on the basis of histologic examination of the CNS. Probability distributions were fitted to serum IFAT results in the EPM+ and EPM-horses, and correlation between serum and CSF results was modeled. Pairs of serum-CSF titers were generated by simulation, and titer-specific likelihood ratios and post-test probabilities of EPM at various pretest probability values were estimated. Post-test probabilities were compared for use of a serum-CSF test combination, a serum test only, and a CSF test only. RESULTS: Post-test probabilities of EPM increased as IFAT serum and CSF titers increased. Post-test probability differences for use of a serum-CSF combination and a serum test only were < or = 19% in 95% of simulations. The largest increases occurred when serum titers were from 40 to 160 and pre-test probabilities were from 5% to 60%. In all simulations, the difference between pre- and post-test probabilities was greater for a CSF test only, compared with a serum test only. CONCLUSIONS AND CLINICAL RELEVANCE: CSF testing after a serum test has limited usefulness in the diagnosis of EPM. A CSF test alone might be used when CSF is required for other procedures. Ruling out other causes of neurologic disease reduces the necessity of additional EPM testing.

Animals↗

PubMed2005

Sample size determination for estimation of the accuracy of two conditionally independent tests in the absence of a gold standard.

We developed an Excel spreadsheet template (available at http://www.epi.ucdavis.edu/diagnostictests/) to calculate sample sizes to estimate sensitivity and specificity with desired precision in the absence of a gold standard. Calculations are predicated on the use of two conditionally independent tests for screening animals from two populations and are based on the methods of Hui and Walter(1980). Sample size calculations rely on asymptotic normality of maximum likelihood (ML) estimates of parameters. Spreadsheets for calculating standard errors for the parameter estimates and for providing ML estimates using cross-tabulated data also are included. An example of application of the methods to bovine paratuberculosis is presented.

Animals↗

PubMed2005

Evaluation of methods for improved detection of Cryptosporidium spp. in mussels (Mytilus californianus).

Bivalve molluscs concentrate Cryptosporidium oocysts from fecal-contaminated aquatic environments and are therefore useful in monitoring water quality. A real-time TaqMan polymerase chain reaction (PCR) system was developed to allow for large scale quantitative detection of Cryptosporidium spp. in mussels (Mytilus californianus). The TaqMan sensitivity and specificity were compared to conventional PCR and direct immunofluorescent antibody (DFA) assays, with and without immunomagnetic separation (IMS), to identify the best method for parasite detection in mussel hemolymph, gill washings and digestive glands. TaqMan PCR and two conventional PCR systems all detected 1 or more oocysts spiked into 1 ml hemolymph samples. The minimum oocyst detection limit in spiked 5 ml gill wash and 1 g digestive gland samples tested by TaqMan PCR and DFA was 100 oocysts, with a 1 log(10) improvement when samples were first processed by IMS. For tank exposed mussels, TaqMan and conventional PCR methods detected C. parvum in <5% of hemolymph samples. No gill washings from these same mussels tested positive by TaqMan PCR or DFA analysis even with IMS concentration. All methods detected the highest prevalence of C. parvum-positive samples in digestive gland tissues of exposed mussels. In conclusion, the most sensitive method for the detection of C. parvum in oocyst-exposed mussels was IMS concentration with DFA detection: 80% of individual and 100% of pooled digestive gland samples tested positive. TaqMan PCR was comparable to conventional PCR for detection of C. parvum oocysts in mussels and additionally allowed for automated testing, high throughput, and semi-quantitative results.

Animals↗

PubMed2005

Factor analysis of a Johne's disease risk assessment questionnaire with evaluation of factor scores and a subset of original questions as predictors of observed clinical paratuberculosis.

Factor analysis was used to examine the interrelationships among 38 variables collected as part of a Johne's disease risk assessment questionnaire completed in 2002 on 815 U.S. dairy operations. Eleven factors were extracted, accounting for two-thirds of the variance encountered in the original variables. Responses to many of the risk assessment questions were closely related. Standardized scores on the 11 factors were calculated for operations providing complete information, and were evaluated as predictors in a model-based logistic regression analysis with the outcome being whether operations had observed one or more cows with clinical signs suggestive of paratuberculosis during the previous year. A logistic regression model was also used to evaluate the predictive ability of a reduced subset of approximately one-third of the original variables that was selected to represent the derived factors. The performance of both sets of predictors was comparable with respect to goodness-of-fit and predictive ability. In conclusion, the length of the current risk assessment instrument could be reduced considerably without a substantial loss of information by removing or combining questions that are strongly correlated.

Animals↗

PubMed2005

Fecal shedding of Mycobacterium avium subsp. paratuberculosis by dairy cows.

Between 1982 and 2000, fecal samples were obtained from 786 cows that were shedding Mycobacterium avium subsp. paratuberculosis (Map). These cows were resident on 93 Pennsylvania dairies (mean herd size, 64 milk cows) that had no or minimal previous testing for Map. Feces were cultured on four tubes of Herrold's egg yolk medium and the distribution of mean Map colony forming units (CFU) was evaluated. Most cows were light (< 10 CFU/tube, 51.4%) or high (> 50 CFU/tube, 30.8%) fecal shedders with fewer cows in the moderate category (10-50 CFU/tube). Of the 786 cows, 192 (24.4%) had colonies in only one of four tubes. In the multivariable negative binomial model, there were significant associations between mean CFU/tube and prevalence, herd size, and season and an interaction between herd size and season. The linear mixed model of continuous tube counts with a random herd effect yielded similar findings with associations with herd size as a continuous variable, season, and an interaction between categorized prevalence and continuous herd size. Variability in CFU/tube was greatest among cows in the same herd, intermediate for replicate tubes from the same cow, and smallest among cows in different herds. Reduction in the number of replicate tubes from four would have reduced the sensitivity of fecal culture for Map by approximately 6% (for three tubes) to 12% (for two tubes).

Animals↗