Seminal fluid was collected by manual ejaculation from 95 dogs. Quantitative aerobic bacterial, qualitative anaerobic bacterial and mycoplasma cultures were performed on the seminal fluid, and their association with presence of inflammatory cells present in the pellet formed after centrifugation of the fluid was investigated. There was a clinically meaningful aerobic bacterial growth in 28.4%, anaerobic bacterial growth in 13.7%, and mycoplasma growth in 57.9% of the seminal fluid samples. Presence of inflammatory cytology was statistically associated with clinically meaningful aerobic bacterial growth. However, of the 78 dogs (82.1%) with clinically meaningful growth of at least one aerobic, anaerobic or mycoplasma organism, 43 (55.1%) had non-inflammatory seminal fluid cytology.

OBJECTIVES:To determine the sensitivity of bacteriologic culture of pooled fecal samples in detecting Mycobacterium paratuberculosis, compared with bacteriologic culture of individual fecal samples in dairy cattle herds.STUDY DESIGN:Cross-sectional study.ANIMALS:24 dairy cattle herds.PROCEDURE:Individual and pooled fecal samples were submitted for bacteriologic culture, and results were compared between these groups.RESULTS:Ninety-four and 88% of pooled fecal samples that contained feces from at least 1 animal with high (mean, > or = 50 colonies/tube) and moderate (mean, 10 to 49 colonies/tube) concentrations of M paratuberculosis, respectively, were identified by use of bacteriologic culture of pooled fecal samples. Prevalences of paratuberculosis determined by bacteriologic culture of pooled and individual fecal samples were highly correlated.CONCLUSIONS AND CLINICAL RELEVANCE:Bacteriologic culture of pooled fecal samples provided a valid and cost-effective method for the detection of M paratuberculosis infection in dairy cattle herds and can be used to estimate prevalence of infection within a herd.

OBJECTIVES:To compare sensitivity of several methods of bacteriologic culture of pooled bovine fecal samples for detection of Mycobacterium paratuberculosis and evaluate homogeneity in number of M paratuberculosis in pooled fecal samples.SAMPLE POPULATION:Feces from 10 dairy cows that shed M paratuberculosis at various concentrations and 1 dairy cow known to be free of infection with M paratuberculosis.PROCEDURE:5 fecal pooling methods, 2 culture methods, and 2 pool sizes were evaluated. Each pooled sample contained 1 infected sample and 4 or 9 uninfected samples.RESULTS:Sensitivity of detection of M paratuberculosis was greater with smaller pool size (5 vs 10 samples/pool). Detection sensitivity was also associated with concentration of bacteria in the infected sample. Results indicated that, compared with concurrent bacterial culture of individual infected samples, 37 to 44% of pooled samples with low bacterial concentrations yielded positive culture results and 94% of pooled samples with high bacterial concentrations yielded positive results.CONCLUSIONS AND CLINICAL RELEVANCE:Bacteriologic culture of pooled fecal samples may provide a valid and cost-effective method of detecting M paratuberculosis infection in cattle herds.

Enterococcus (Streptococcus) durans is a motile, Grampositive coccus found in the alimentary tract of animals and in water, soil, sewage, and vegetati~n.~.~ Enteric disease and diarrhea were first associated with E. durans when Grampositive cocci were seen adhered to enterocytes in the small intestine of diarrheic pigs’ and foals.Io E. durans subsequently was shown to induce diarrhea in gnotobiotic pigs7Jo and conventionally reared foalsI0 challenged with strains of porcine or equine origin. Diarrhea of 8 hours duration occurred in eight of ten, 1 1 day-old Plott hound pups from a private residence. The bitch had been vaccinated against distemper, hepatitis, and parvovirus and did not have clinical signs. Despite intensive fluid and antimicrobial therapy, three pups died, and one of these was presented dead for necropsy. At necropsy, the pup was in excellent flesh, moderately dehydrated, and had severe hyperemia of the small intestinal wall from the duodenum through the jejunum. Scant amounts of liquid brown feces were present in the large intestine. Kidney and jejunum were fixed in 10% neutral-buffered formalin, embedded in paraffin, sectioned, and examined after staining with hematoxylin and eosin and Gram’s stains. Pieces (0.5 cmz) of formalin-fixed small intestine were immersed in 3% glutaraldehyde in 0.1 M sodium phosphate buffer at pH 7.2 and refrigerated at 4 C until processed for transmission and scanning electron microscopic examinations by procedures described previ~usly.~ Portions of small intestine were inoculated onto sheep blood, brilliant green, MacConkey and cholistin naladixic acid agars, and incubated aerobically at 37 C. Feces were processed and negatively stained using procedures described previously8 and examined using an electron microscope. Microscopically, villi in the jejunum were long, slender, and densely colonized by Gram-positive coccoid bacteria (Figs. 1, 2). Bacterial colonization of enterocytes extended from the tips to the base of villi. The morphologic structure of enterocytes in areas with bacterial adherence was unaltered, except for changes attributed to post-mortem decomposition. Intestinal crypts were orderly in arrangement and structure, although a few crypts were dilated and contained neutrophils. Blood vessels in the submucosa, tunica muscularis, and serosa were dilated and blood-filled. The lumens of venules in the lamina propria were sometimes occluded by fibrinous thrombi. A few neutrophils were admixed with coccoid and rod bacteria in the jejunal lumen; but in general, inflammation was mild. Ultrastructurally, coccoid bacteria were closely apposed to enterocytes (Fig. 3). Fine filaments consistent in morphologic structure with fimbriae extended from bacterial cell walls to adjacent bacteria and to enterocyte brush borders (Fig. 3). Swelling of cytoplasmic organelles and distortion of microvilli most likely resulted from post-mortem decomposition. Intracellular cocci were frequently seen within lysozomes of villous epithelial cells; but the morphologic integrity of intracytoplasmic cocci was rarely intact. In most instances intracytoplasmic cocci had been digested, leaving finely granular, irregularly shaped, osmiophilic deposits within lysosomes (Fig. 4). A mixed growth of non-hemolytic Escherichia coli and Enterococcus sp. was isolated from the jejunum. The enterococcus isolate, further characterized using a commercial mi-