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This research study was conducted to determine if bird depredation in feedlots is associated with the prevalence of ciprofloxacin-resistant Escherichia coli in cattle and to determine if removal of invasive bird species could be an effective management strategy to help reduce ciprofloxacin-resistant E. coli in cattle within the United States. European starlings (Sturnus vulgaris) were collected from feedlots within multiple geographic regions within the United States and European starlings within all regions tested positive for ciprofloxacin-resistant E. coli, but prevalence differed by region. Total number of birds on feedlots were positively associated with increased cattle fecal shedding of ciprofloxacin-resistant E. coli. Targeted control of invasive European starlings reduced bird numbers on feedlots by 70.4%, but decreasing populations of European starlings was not associated with corresponding reductions in bovine fecal prevalence of ciprofloxacin-resistant E. coli. These data provide evidence for the role of wild bird depredation in feedlots contributing to fecal shedding of ciprofloxacin-resistant E. coli, but a single month of European starling control in feedlots was not sufficient to impact the fecal carriage of this organism in cattle.

Salmonella and Campylobacter cause a significant number of human illnesses globally, most of which are food related. Cattle can be asymptomatic carriers of both of these pathogens. The objective of this study was to determine the association between the concentration of Salmonella and Campylobacter pre- and postharvest in cattle. Samples were collected from each of 98 individually identified cattle during the periharvest and postharvest period. For each animal, four different phases were sampled: on farm (fecal sample), poststunning and exsanguination (hide sponge and rectal content sample [lairage]), prechilling (carcass sponge), and final product (ground meat). Salmonella and Campylobacter were cultured and quantified at each stage by using the direct dilution and most probable number (MPN) method. Salmonella was not isolated from any sample. The proportion (%) of samples that were Campylobacter positive was 77, 82, 97, 55, and 12 for farm, rectal content, hide, carcass, and meat samples respectively. The mean Campylobacter concentration for each sample was as follows: fecal sample from farm, 3.7×10(4) cfu/g; rectal content sample from lairage, 1.6×10(5) cfu/g; hide sponge, 0.9 cfu/cm(2); carcass sponge, 8.7 cfu/half carcass; and meat, 1.1 cfu/g. There were no associations between Campylobacter concentrations for any two sample types. This lack of association could indicate that there is an environmental reservoir that can contaminate the final meat product, or since the majority of animals were positive entering the slaughter process, that the process itself reduces the load of Campylobacter regardless of the initial concentration. In addition, contamination of beef may be more strongly associated with periharvest practices than animal carriage rates.

CTX-M extended-spectrum β-lactamases are enzymes produced by bacteria that are capable of inhibiting the antimicrobial effects of cephalosporin drugs. Recently, the first domestically acquired Salmonella in the United States expressing bla(CTX-M) was reported. This is a concern because expanded-spectrum cephalosporins are the treatment of choice for invasive Gram-negative infections, including salmonellosis in children. Because Salmonella transmission is primarily foodborne, there is also concern that resistant enteric bacteria from livestock can be transferred through the food supply chain to consumers. bla(CTX-M) has not been previously identified in bacterial isolates from food animal populations in the United States. We report the recovery of CTX-M-type extended-spectrum β-lactamases from fecal Escherichia coli of sick and healthy dairy cattle in Ohio. Four individual fecal samples yielded E. coli isolates representing three clonal strains that carried bla(CTX-M) on transferable plasmids. Two distinguishable plasmids were identified, each encoding bla(CTX-M-1) or bla(CTX-M-79). Transferrable bla(CTX-M) genes in bovine E. coli have the potential to serve as a reservoir of resistance for pathogens and may represent a public health concern.

Objective: To estimate the efficacy of a commercially available Salmonella enterica subunit vaccine on the subclinical shedding of S enterica in dairy cattle., Design: Randomized, controlled trial., Animals: 175 mature cows on 2 dairy farms with a history of S enterica infection., Procedures: 25% of the mature cows from each herd were systematically randomized to receive an S enterica subunit vaccine following label guidelines. The remaining 75% of cows in each herd served as nonvaccinated controls. Fecal samples were collected from all cows at the time of initial vaccination (day 0), booster vaccination (day 14), 2 weeks following the booster vaccination (day 28), and 10 weeks following the start of the trial (day 70). All samples were processed on the day of collection and cultured for S enterica., Results: 651 fecal samples were obtained over the entire study period. Salmonella enterica was recovered from 46 (7.1%) of the samples. Shedding of S enterica was similar for vaccinated and nonvaccinated control cows on each of the collection dates., Conclusions and Clinical Relevance: The study revealed no evidence that extralabel vaccination with a commercial subunit S enterica vaccine reduced shedding of S enterica in subclinically infected dairy cows in these herds.

Objective: To quantify effects of certified health programs on the sale price of beef calves sold through a livestock videotape auction service., Design: Longitudinal study., Sample Population: 26,502 lots representing 3,205,192 beef calves sold through a livestock videotape auction service between 1995 and 2005., Procedures: Data describing each lot of beef calves that were marketed from 1995 through 2005 by a livestock videotape auction service were obtained from sale catalogues. For each year of the study, multiple regression analysis was used to quantify the effect of certified health programs on sale price., Results: For each year of the study, beef calves that qualified for the 2 most intensive certified health programs sold for significantly higher prices, compared with prices for similar calves that were not in a certified health program, had not been vaccinated against respiratory tract viruses, and were not weaned before delivery. Price premiums for calves in the most intensive certified health program ranged from $2.47/100 lb (hundredweight [cwt]; 1 cwt equals 45.45 kg) in 1995 to $7.91/cwt in 2004. Price premiums paid for calves qualifying for the next most intensive certified health program ranged from $0.99/cwt in 1996 to $3.47/cwt in 2004. The percentage of the total number of lots in the 2 most intensive certified health programs increased over time., Conclusions and Clinical Relevance: Findings from this study indicated that implementation of the 2 most intensive certified health programs consistently increased the price of beef calves, and these price premiums increased over time.

Objective: To compare shedding patterns and serologic responses to bovine coronavirus (BCV) in feedlot calves shipped from a single ranch in New Mexico (NM calves) versus calves assembled from local sale barns in Arkansas (AR calves) and to evaluate the role of BCV on disease and performance., Animals: 103 feedlot calves from New Mexico and 100 from Arkansas., Procedures: Calves were studied from before shipping to 35 days after arrival at the feedlot. Nasal swab specimens, fecal samples, and serum samples were obtained before shipping, at arrival, and periodically thereafter. Bovine coronavirus antigen and antibodies were detected by use of an ELISA., Results: NM calves had a high geometric mean titer for BCV antibody at arrival (GMT, 1,928); only 2% shed BCV in nasal secretions and 1% in feces. In contrast, AR calves had low antibody titers against BCV at arrival (GMT, 102) and 64% shed BCV in nasal secretions and 65% in feces. Detection of BCV in nasal secretions preceded detection in feces before shipping AR calves, but at arrival, 73% of AR calves were shedding BCV in nasal secretions and feces. Bovine coronavirus infection was significantly associated with respiratory tract disease and decreased growth performance in AR calves., Conclusions and Clinical Relevance: Replication and shedding of BCV may start in the upper respiratory tract and spread to the gastrointestinal tract. Vaccination of calves against BCV before shipping to feedlots may provide protection against BCV infection and its effects with other pathogens in the induction of respiratory tract disease.

Escherichia coli O157:H7 (EC O157) is an important cause of foodborne disease. Cattle are reservoirs for the bacteria and are implicated in transmission to humans. Prevalence data in prefeedlot calves are limited. With the use of sensitive methods, a cohort of weaned beef calves (n = 408) was sampled before and after preconditioning to estimate fecal point prevalence and describe changes in EC O157 fecal shedding. EC O157 isolates were confirmed and characterized by PCR and pulsed-field gel electrophoresis. Calves from 29 cow-calf farms were commingled at three preconditioning sites and placed on a transition ration containing oxytetracycline (200 g/ton) for 45 days. Initial animal-level fecal point prevalence was 2.5% (95% confidence interval, 1 to 5) with a herd-level prevalence of 17.2% (95% confidence interval, 6 to 36). Point prevalence following the preconditioning feeding period was 0%. An unexpected finding in our study was EC O157 isolates that were Shiga toxin-deficient. Pulsed-field gel electrophoresis subtypes of EC O157 were unique in epidemiologically unlinked herds, except one herd that had two unique subtypes. We expected, but observed, neither increased fecal shedding in the cohort nor horizontal transmission of unique EC O157 subtypes. The absence of fecal shedding following the 45-day feeding period might be attributable to seasonal influences, inhibitory concentrations of oxytetracycline in the transition ration, or transient colonization that ended before sampling. EC O157 is apparently widely dispersed at low prevalence in U.S. prefeedlot, weaned calves.

The purpose of our mail survey was to compare the adoption of management practices recommended for Johne's disease (JD) control between herds involved in whole-herd testing programs versus those that do not routinely test the entire herd for JD. A questionnaire consisted of 38 closed-ended questions that inquired about: general herd characteristics; management practices related to JD control; changes that occurred within the last 5 years regarding management practices recommended for the control of JD; producer knowledge of JD; the perceived infection status of the herd by the producer; and herd JD-testing history. The questionnaire was mailed to 810 Ohio dairy producers in September 2002; 266 questionnaires were returned (32.8% response). We used univariable logistic-regression models to assess the relationship between whole-herd testing status (TESTING versus NON-TESTING) and each management practice, each change in management practice and producer knowledge about JD. Because it is conceivable that only producers who believe their herds to be infected would be motivated to adopt the management practices recommended for control of JD, the comparisons were repeated with models that controlled for producer-perceived infection status. Of the 20 management practices recommended for JD control that we evaluated, 7 differed between TESTING and NON-TESTING herds. Additionally, TESTING herds more-frequently reported adopting changes within the past 5 years relative to NON-TESTING herds with respect to 7 of 9 management practices evaluated. Producers with TESTING herds also reported greater familiarity with JD than those with NON-TESTING herds.

Short-term stability of measurements of antimicrobial susceptibility of Escherichia coli isolated from feces of feedlot cattle is important in developing monitoring and surveillance programs. Frequent evaluations (i.e., daily) are resource intensive and in some situations may be impractical for long-term sampling protocols. Consequently, a point-in-time measurement will need to be used to represent conditions in the perisampling period. In this study, 30 fecal samples were collected from each of 6 cattle pens on a commercial cattle feedlot on 2 occasions separated by 48 hours. Escherichia coli was isolated from single and pooled samples. The isolates were tested for antimicrobial susceptibility against a panel of 17 antimicrobials. Resistance to 5 antimicrobials (ampicillin, nalidixic acid, streptomycin, sulfamethoxazole, and tetracycline) was detected in single and pooled samples from both sampling periods (days 1 and 3). The prevalence of isolates resistant to these 5 antimicrobials was 2% or higher in all treatment combinations except for pools obtained from day 3 samples. Lower levels of resistance to 6 more antimicrobials were detected inconsistently across the single and pooled samples. Logistic models constructed for the antimicrobials to which the E. coli isolates were most commonly resistant demonstrated that there were no significant differences between periods (P > 0.10) and between single and pooled samples (P > 0.20). The distribution of the number of antimicrobials to which isolates were resistant was consistent for the single samples across periods, but there appeared to be a lower prevalence of any resistance in day 1 pooled samples. A larger number of resistant phenotypes were detected in the single samples than in the pooled samples, and resistant phenotypes with prevalence of less than 2% were detected inconsistently across periods and single and pooled samples. Resistance to individual antimicrobials was consistent by all measures when the prevalence was at least 2%. Inconsistent results were obtained for antimicrobials to which resistance rarely occurred. The apparent inconsistencies do not appear to be related to external factors but rather to sampling intensity. Short-term stability is a plausible assumption under sampling strategies that are designed to detect specific levels of prevalence. However, when resistance levels fall below these levels, there will likely be fluctuations in the presence or absence of rare resistant phenotypes and in their prevalence and central tendency measures.

The objectives of this study were to determine the prevalence of bovine torovirus (BoTV) in bovine fecal samples from diarrhea cases submitted to the Ohio Animal Disease Diagnostic Laboratory (ADDL) and to assess if a relationship exists between BoTV and the other enteric pathogens detected. From November 1999 to May 2001, 259 specimens from 53 calves (< or = 6 months old), 27 young adults (52 years), 125 adults (> or = 2 years), and 54 animals of unknown age were examined by an antigen-capture enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase-polymerase chain reaction (RT-PCR) assay developed to detect BoTV. Testing for other enteric pathogens was performed by ADDL, and the results were analyzed with the BoTV data. The BoTV was detected using ELISA or RT-PCR in 9.7% (25/259) of the clinical samples, 56% (14/25) of which were from calves (P < 0.001) representing 26.4% (14/53) of the calves tested. Of the BoTV-positive calves, 71% (10/14) were less than 3 weeks of age. In 11/25 positive specimens, BoTV was the only pathogen detected among those examined. Other enteric organisms detected alone or in combination with BoTV in calf samples were rotavirus, coronavirus, Salmonella spp., Cryptosporidium spp., and Giardia spp.; but no consistent association between BoTV and these organisms was observed. In summary, BoTV was detected in fecal samples from cattle with diarrhea, principally in young calves less than 3 weeks of age. Future studies of infectious diarrhea in cattle should also include assays for this etiologic agent.

Objective: To determine the prevalence, fecal shedding pattern, and association of bovine torovirus (BoTV) with diarrhea in veal calves at time of arrival and periodically throughout the first 35 days after their arrival on a veal farm., Animals: 62 veal calves., Procedure: Fecal samples collected on days 0, 4, 14, and 35 after arrival were tested for BoTV by use of ELISA and reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Paired serum samples obtained from blood collected on days 0 and 35 were analyzed for BoTV antibodies with a hemagglutination inhibition assay. Fecal samples were also screened for other enteric pathogens, including rotavirus, coronavirus, and Cryptosporidium spp., Results: Fecal shedding of BoTV was detected in 15 of 62 (24%) calves by use of ELISA and RT-PCR assay, with peak shedding on day 4. A significant independent association between BoTV shedding and diarrhea was observed. In addition, calves shedding > or = 2 enteric pathogens were more likely to have diarrhea than calves shedding < or = 1 pathogen. Calves that were seronegative or had low antibody titers against BoTV (< or = 1:10 hemagglutination inhibition units) at arrival seroconverted to BoTV (> 4-fold increase in titer); these calves were more likely to shed virus than calves that were seropositive against BoTV at arrival., Conclusions and Clinical Relevance: Shedding of BoTV was strongly associated with diarrhea in neonatal veal calves during the first week after arrival at the farm. These data provide evidence that BoTV is an important pathogen of neonatal veal calves.

Factor analysis was used to assess relationships in the minimum-inhibitory concentration among 17 antimicrobials tested on isolates of Escherichia coli isolated from 360 faecal samples obtained from feedlot cattle. Six factors were extracted using maximum-likelihood factor analysis. The factors were interpretable antimicrobial groupings based on class of antimicrobial and previously described associations. New-generation cephalosporins, older-generation beta-lactams, fluoroquinolones and aminoglycosides grouped separately as classes of antimicrobials on four of the six factors. One of the remaining factors was a grouping of antimicrobials that had been identified as being related in previous feedlot studies. The last factor was a grouping of three of the five antimicrobials that comprise the antimicrobials found in penta-resistant strains of Salmonella Typhimurium. The factor analysis described patterns in the MIC data that would not have been apparent if only antimicrobial-resistance data categorized as susceptible-resistance had been analysed.

Objective: To evaluate the effect of feeding aspirin and supplemental vitamin E on growth performance, lung lesions, plasma concentrations of 3-methylindole (3MI), and 3-methyleneindolenine (3MEIN)-adduct concentrations in blood and pulmonary tissues of feedlot cattle., Animals: 256 crossbred steers; 64 cattle were used in experiment 1 and 192 cattle were used in experiment 2., Procedures: A 2 X 2 factorial design was used for each experiment. Treatment factors were aspirin (0 or 3 g daily) and vitamin E (200 or 1,500 IU daily). Steers were housed in pens (8 steers/pen). Steers were slaughtered on days 59 and 138 for experiments 1 and 2, respectively. Lungs were grossly evaluated. 3MEIN-adduct concentrations were determined, and blood and pulmonary tissues., Results: Treatment was not associated with improvement or adverse effects on weight gain, dry-matter intake, or feed efficiency in experiment 2. In experiment 1, 36 of 63 (57.1%) steers had lung lesions. Lesions were not associated with treatment or concentrations of 3MI and 3MEIN-adduct. Plasma 3MI concentration and concentrations of 3MEIN-adduct in blood and pulmonary tissues were 3.11 microg/mL, 0.51 U/microg of protein, and 0.49 U/microg of protein, respectively. Aspirin was associated with increased blood concentrations of 3MEIN-adduct for diets that did not contain supplemental vitamin E., Conclusions and Clinical Relevance: Differences n performance of feedlot steers were not associated with treatment diet. It is possible that concurrent exposure of feedlot cattle to other factors typically associated with development of respiratory tract disease would affect these findings.

Recently, bovine coronavirus (BCV) has been isolated from new cattle arrivals to feedlots, but the association between respiratory and enteric infections with BCV in feedlot cattle remains uncertain. Fecal and nasal swab samples from 85 Ohio Agricultural Research and Development Center (OARDC) feedlot cattle averaging 7 months of age were collected at arrival (0) and at 4, 7, 14, and 21 days postarrival (DPA). An antigen capture enzyme-linked immunosorbent assay (ELISA) was used to detect concurrent shedding of BCV in fecal and nasal samples. All samples ELISA positive for BCV were matched with an equal number of BCV ELISA-negative samples and analyzed by reverse transcription-polymerase chain reaction (RT-PCR) of the N gene. Paired sera were collected at arrival and 21 DPA and tested for antibodies to BCV using an indirect ELISA. Information on clinical signs, treatments provided, and cattle weights were collected. The overall rates of BCV nasal and fecal shedding were 48% (41/85) and 53% (45/85) by ELISA and 84% (71/85) and 96% (82/85) by RT-PCR, respectively. The peak of BCV nasal and fecal shedding occurred at 4 DPA. Thirty-two cattle (38%) showed concurrent enteric and nasal shedding detected by both tests. Eleven percent of cattle had antibody titers against BCV at 0 DPA and 91% of cattle seroconverted to BCV by 21 DPA. The BCV fecal and nasal shedding detected by ELISA and RT-PCR were statistically correlated with ELISA antibody seroconversion (P < 0.0001); however, BCV fecal and nasal shedding were not significantly related to clinical signs. Seroconversion to BCV was inversely related to average daily weight gains (P < 0.06). Twenty-eight respiratory and 7 enteric BCV strains were isolated from nasal and fetal samples of 32 cattle in HRT-18 cell cultures. These findings confirm the presence of enteric and respiratory BCV infections in feedlot calves. Further studies are needed to elucidate the differences between enteric and respiratory strains of BCV and their role in the bovine respiratory disease complex of feedlot cattle.

Objective: To assess fecal and nasal shedding patterns of bovine torovirus (BoTV) in cattle at time of arrival and periodically throughout the first 21 days after arrival at a feedlot., Animals: 57 steers., Procedure: Fecal and nasal-swab samples collected on days 0, 4, 14, and 21 after arrival were tested for BoTV, using ELISA. A subset of samples from calves testing positive and negative for BoTV was analyzed, using reverse transcriptase-polymerase chain reaction (RT-PCR). Paired serum samples were collected on days 0 and 21 and tested for BoTV antibodies, using a hemagglutination inhibition assay., Results: Overall rate of fecal shedding of BoTV was 21 of 57 (37%) by ELISA and 40 of 42 (95%) by RT-PCR with peak shedding on day 4. Diarrhea was more common in calves shedding BoTV than those not shedding the virus (odds ratio, 1.72). Overall rate of nasal shedding of BoTV was 15 of 57 (26%) by ELISA and 42 of 42 (100%) by RT-PCR, with peak shedding on day 0. Specificity of the RT-PCR product was confirmed by sequence analysis. Approximately 93% of the calves seroconverted to BoTV (> 4-fold increase in titer). Differences were not detected between calves shedding BoTV and nonshedders in relation to disease and treatments, perhaps because of the low number of cattle in the study., Conclusions and Clinical Relevance: This study confirmed BoTV infections in feedlot cattle, including BoTV antigen and viral RNA in nasal secretions, and the shedding pattern during the first 21 days after arrival in a feedlot.

Objective: To estimate prevalence of Salmonella spp in Ohio dairy farms and to identify potential risk factors for fecal shedding of salmonellae., Design: Cross-sectional study., Sample Population: 105 Ohio dairy farms., Procedure: Individual fecal samples from all mature cows in study herds were tested for Salmonella spp by use of standard bacteriologic culture procedures. Herds were identified as infected if at least 1 cow was shedding Salmonella spp. Information regarding herd characteristics, management practices, and health history were collected. Potential risk factors for herd-level Salmonella infection were identified., Results: In 31% of the study herds (95% confidence interval, 22 to 40%), at least 1 cow was shedding Salmonella spp. Six percent of 7,776 fecal samples contained Salmonella organisms; prevalence within infected herds ranged from < 1 to 97%. Herd size, use of free stalls for lactating and nonlactating cows, and use of straw bedding in nonlactating cows were significantly associated with fecal shedding of Salmonella spp, as determined by use of univariate analysis. By use of multivariate analysis, large herds were more likely to be infected than smaller herds; however, no other factors were associated with Salmonella infection after adjustment for herd size., Conclusions and Clinical Relevance: Subclinical shedding of Salmonella spp is common in Ohio dairy herds, although we could not identify specific interventions that may influence the prevalence of Salmonella spp on dairy farms. It appears that large herd size and intensive management may provide an environment conducive to Salmonella shedding and chronic dairy herd infection.

Objective: To monitor patterns of Salmonella fecal shedding in naturally infected dairy herds, determine the association between fecal shedding and individual animal production measures, and evaluate potential risk factors for shedding of Salmonella organisms among cattle in dairy herds., Design: Longitudinal study., Sample Population: 5 Ohio dairy herds., Procedure: For 3 herds, fecal samples were collected from all mature cows and unweaned calves 7 times during an 18-month period. For the remaining 2 herds, fecal samples were collected from 50 lactating cows 6 times during a 12-month period. Individual animal production records for 3 herds were used to examine associations between individual fecal Salmonella shedding status and 305-day mature-equivalent milk production, somatic cell count, milk fat content, and milk protein content. Multivariable logistic regression was used to test for associations between fecal shedding status and breed, lactation status, lactation number, and duration of lactation., Results: None of the adult animals had clinical signs of salmonellosis, but prevalence of fecal Salmonella shedding at individual collection times ranged from 0 to 99% for cows and from 0 to 67% for unweaned calves. Mature cows were more likely to be shedding Salmonella organisms than were unweaned calves. Within herds, lactation status and duration of lactation for individual animals were associated with Salmonella shedding status. Salmonella fecal shedding status was not associated with individual cow production measures., Conclusions and Clinical Relevance: Results suggest that subclinical fecal Salmonella shedding can persist in dairy herds for up to 18 months with no measurable effects on health or production of individual cows.

The combination of medium and growth conditions, including transport enrichment medium (TEM), transport time, TEM incubation time, and growth medium, that best support Campylobacter fetus subsp. venerealis while inhibiting contaminants was studied. The 3 TEMs evaluated, Weybridge, Cary-Blair, and 0.85% saline solution, were inoculated with preputial smegma spiked with C. fetus subsp. venerealis and held in the laboratory for 4 or 24 hours before inoculation onto growth medium. The effect of overnight incubation at 37 C of the TEM was also evaluated. Median scores of C. fetus subsp. venerealis growth and microbial contaminant inhibition were compared within TEM, transport time, overnight incubation, and growth medium groups using the Mann-Whitney U-test and the Kruskal-Wallis test. The proportion of samples with any growth or contamination within each group was also compared using the chi-square test. Results suggest that the growth of C. fetus subsp. venerealis was influenced by 3 of the 4 criteria evaluated. Weybridge TEM more effectively maintained the organism than did either Cary-Blair or 0.85% saline solution (P < 0.001). Transport time of 4 hours rather than 24 hours (P < 0.001) and avoiding overnight incubation of TEM at 37 C (P < 0.001) were associated with improved growth. Significant differences were not identified among growth media; however, Skirrow Campylobacter agar appeared to yield slightly better growth than did either blood agar or Greenbriar Plus agar. Contaminant growth was also influenced by 3 of the 4 variables. Weybridge TEM inhibited contaminant growth more effectively than did either Cary-Blair or 0.85% saline solution (P < 0.001). Transport time was not associated with contaminant growth. Eliminating overnight incubation of TEM reduced contamination (P < 0.01). Skirrow agar was preferred to both blood agar and Greenbriar Plus agar for suppression of contaminants on solid medium (P < 0.001). These results suggest that the detection of C. fetus subsp. venerealis is enhanced when preputial smegma samples arrive at the diagnostic laboratory within 4 hours after inoculation into Weybridge TEM and are transferred to Skirrow agar the same day they arrive in the laboratory.