Effect of Transport Enrichment Medium, Transport Time, and Growth Medium on the Detection of Campylobacter Fetus subsp. Venerealis

Abstract

Abstract. 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 Ͻ 0.001). Transport time of 4 hours rather than 24 hours (P Ͻ 0.001) and avoiding overnight incubation of TEM at 37 C (P Ͻ 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 Ͻ 0.001). Transport time was not associated with contaminant growth. Eliminating overnight incubation of TEM reduced contamination (P Ͻ 0.01). Skirrow agar was preferred to both blood agar and Greenbriar Plus agar for suppression of contaminants on solid medium (P Ͻ 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.Bovine genital campylobacteriosis is a contagious venereal disease caused by Campylobacter fetus subsp. venerealis. This gram-negative, microaerophilic, motile, spiral-shaped rod is harbored in the proximal preputial cavity of bulls and in the genital tract of cows. 3 Transmission of the bacterium occurs mechanically between cows and bulls during coitus, by contaminated semen used for artificial insemination, or between bulls through contact with either contaminated bedding or contaminated semen-collection equipment. 1,6 Campylobacteriosis in the cow or heifer causes irregular estrous cycles, uterine infections that prevent conception, and early embryonic death. 1,6,8 Most cows recover from infection and begin estrous cycles after several months, but the herd experiences a decreased pregnancy rate and a prolonged ca...

Comparison of Culture and a Novel 5′ Taq Nuclease Assay for Direct Detection of Campylobacter fetus subsp. venerealis in Clinical Specimens from Cattle

“…The disease is caused by Campylobacter fetus subsp. venerealis and is spread by infected bulls during servicing, by contaminated semen, or between bulls (41). Campylobacteriosis is an Office International des Epizooties list B notifiable disease that is considered to have socioeconomic and/or public health implications and is thus significant in the international trade of animals and animal products.…”

“…Several methods for the collection of preputial smegma and vaginal mucus have been investigated in order to improve the reliability of selective culture-based diagnostic procedures or ELISA. These procedures have included preputial washes, scrapes (55), mucus swabs, blotting (23), and the commonly used scrape/aspiration methods with a pipette (26,41,55). The use of swabs and blotting has been limited to female cattle.…”

Validation of a Monoclonal Antibody-Based Capture Enzyme-Linked Immunosorbent Assay for Detection of Campylobacter fetus

“…Of the 66 samples analyzed, 65 were positive by ELISA but only 49 were positive by culture. Although the method used for the culture of C. fetus subspecies is reliable, the inoculum size, the presence of competing microflora, and the environmental conditions that affect the TEM vials during transport are known to influence the success of culture (9,21). As such, two TEM samples are routinely inoculated for every sample in Canada.…”

Simple media and conditions for inter-laboratory transport ofCampylobacter jejuniisolates

“…can be kept at À80 C for many years but especially air carriage in dry ice or under liquid nitrogen is rather expensive. The transport of isolates of Campylobacter -a true microaerophilic bacteriumunder aerobic conditions is often unsuccessful (Wang et al 1980;Luechtefeld et al 1981;Wasfy et al 1995;Monke et al 2002;Tresierra Ayala et al 2006).…”