The prevalence and intensity of Cryptosporidium infection were examined in 445 Holstein calves at 10 dairy farms in western Washington, near Seattle. Fifty-one percent (176) of calves in the 7- to 21-day-old age group (n = 342) were positive for oocysts in the feces by carbolfuchsin staining. Prevalence and intensity of infection were highest in calves 8 to 14 days old; prevalence was 60% in this group, and 48% of the Cryptosporidium-positive calves had oocyst shedding at a 4+ level. A seasonal pattern in prevalence was not evident.

Thirty-two isolates of Campylobacter fetus subsp venerealis were obtained from 1 bull and 4 heifers with experimentally induced infection. When whole-cell antigens of isolates were cross titrated with antisera to the infecting strain, isolates from 3 heifers had limited antigenic variation, whereas whole-cell antigens of isolates from 2 cattle (the bull and a heifer) differed serologically from those of the infecting strain. Changes were detected specifically in 6 heat-labile antigens. Of the 6 heat-labile factors evaluated, all were initially present on the infecting parent strain, but not on early isolates obtained from 4 of the 5 cattle. Restriction enzyme analysis revealed minor variation in the DNA fingerprints of isolates obtained from individual cattle, thus implying stability of the Campylobacter genome once persistent infection is established. Isolates with identical restriction enzyme patterns expressed different heat-labile antigens. Correlation could not be found between the DNA electrophoretic pattern and the expression of heat-labile antigens.

The minimal inhibitory concentrations (MIC) of sulfonamides were determined against Bordetella bronchiseptica (n = 10), Pasteurella multocida (n = 10), Haemophilus pleuropneumoniae (n = 20), and Streptococcus suis (n = 10) strains isolated from pigs with atrophic rhinitis, pneumonia, or meningitis. Sulfonamides tested in an agar dilution method were sulfachloropyridazine, sulfadiazine, sulfadimethoxine, sulfamethazine, sulfadoxine, sulfisoxazole, sulfamerazine, sulfamethoxazole, sulfamethoxypyridazine, sulfanilamide, sulfatroxazole, and sulfisomidine. Results indicated that monotherapy of S suis infections with sulfonamides should not be encouraged because the MIC50 of all sulfonamides investigated was greater than 32 microgram/ml. The MIC50 of the sulfonamides against B bronchiseptica ranged from 0.5 to 8 migrogram/ml, against P multocida from 2 to 32 microgram/ml, and against H pleuropneumoniae from 8 to 64 microgram/ml. The MIC50 of sulfachloropyridazine, sulfadiazine, sulfadimethoxine, sulfamerazine, and sulfamethoxazole for the gram-negative bacteria did not exceed 16 microgram/ml. Among these compounds, sulfamethoxazole had the highest activity. The frequently prescribed sulfamethazine had an overall low antimicrobial activity.

Eight healthy nonstressed calves were inoculated with Pasteurella haemolytica serotype 1, by instilling a broth culture into the middle nasal meatus of the left nostril. The inoculated left nostrils shed P haemolytica from the ventral nasal meatus at a steady rate for a mean of 7 days, whereas the uninoculated right nostrils of the same calves shed P haemolytica sporadically and in lower concentrations. The duration, frequency, and concentration of P haemolytica shed from the inoculated nostrils was significantly (P less than 0.05) greater than from the nostrils of other healthy calves that had been exposed by instilling the culture into the ventral nasal meatus of both nostrils in a previous study. The concentration of antibodies (IgG, IgA, and IgM) to P haemolytica increased significantly (P less than 0.05) in serum and nasal secretions after exposure. Four weeks after initial P haemolytica exposure, calves were exposed to infectious bovine rhinotracheitis virus and became clinically ill. Four calves were induced to shed P haemolytica from both nostrils by the virus infection; thus, they were harboring the bacterium and were susceptible to active recolonization. Four calves were not induced to shed P haemolytica. The apparent reason was not that they were resistant to active colonization, but that they were no longer harboring the bacterium, because they became active shedders after they were reinfected with P haemolytica.

Salivary glands from males of 3 Dermacentor species (D andersoni, D variabilis and D occidentalis) that were infected with either the Virginia or Idaho isolate of Anaplasma marginale as nymphs or adults were examined for colonies of A marginale by use of light and electron microscopy. Prior to dissection of salivary glands, exposed ticks were held at 25 C for 15 to 18 days, followed by a 3-day incubation at 37 C. Ticks of 2 species transmitted A marginale to calves; the third tick species was confirmed infected by demonstration of typical colonies in tick gut cells, but transmission was not attempted; Colonies of A marginale were seen with light microscopy in salivary glands of all 3 species of ticks; they were located in acinar cells that contained simple granules. Colonies varied morphologically from small, compact ones to larger structures that contained distinct organisms and often were adjacent to the host cell nucleus. Electron microscopy confirmed that the colonies were rickettsial organisms. Morphologic features of A marginale varied and included reticulated forms, forms with electron-dense centers, and small particles; these various forms were similar to those described previously in midgut epithelial cells of ticks. We believe that the organism seen within tick salivary glands may replicate in the glands before its transmission to the vertebrate host.

To determine their capacity to host Brucella abortus, face flies were examined 1 to 120 hours after feeding on broth containing bacteria and bovine erythrocytes. Brucella abortus was cultured in large numbers from whole flies for 12 hours after feeding, but not after 72 hours. Histologic analysis showed that brucellae were rapidly taken into the midgut, sequestered from erythrocytes, transiently stored, and shed in the feces; there was no evidence of bacterial replication within epithelial cells. Immunoperoxidase and immunogold techniques revealed that most brucellae in the gut were confined to the lumen by the peritrophic membrane, that brucellae were degraded in secondary lysosomes of midgut epithelial cells, and that intact brucellae passed into connective tissues surrounding the midgut. Bacterial excretion without midgut replication is consistent with transient, but not long-term, insect transmission in nature.

Experiments were conducted to establish a persistent Salmonella typhimurium infection in convalescent swine, and to determine rate of shedding and distribution of the organism in internal organs. Naturally farrowed Salmonella-free pigs (n = 37) were orally exposed to S typhimurium when 7 to 8 weeks old. Fecal samples, tonsillar scrapings, and rectal swab specimens were examined bacteriologically for S typhimurium at weekly intervals after exposure until necropsy (maximum of 28 weeks after exposure). Necropsies of 1 to 4 randomly selected pigs were conducted at 2, 4, and 7 days and at 2, 4, 6, 8, 12, 16, 20, 24, and 28 weeks after exposure. The following internal organs were examined bacteriologically for S typhimurium: liver, spleen, kidney, gallbladder, heart, lung, and stomach; segments of the intestinal tract with corresponding lymph nodes; lymph nodes from lymphocenters of the head and neck, thoracic and abdominal cavities, pelvic wall, and thoracic and pelvic limbs. Fecal samples were 83 to 100% culture-positive up to postexposure (PE) week 22, then varied from 14 to 67% positive until PE week 28. At least 60% of tonsillar swab specimens and 50% of rectal swab specimens were culture-positive up to PE week 20, after which they varied from 0 to 70% positive until PE week 28. At necropsy, S typhimurium was recovered most freguently from tonsils (93.5% positive), followed by segments of the intestinal tract from caudal portion of jejunum to rectum (71% recovery from cecum), and mandibular (54.8%) and ileocolic (45.2%) lymph nodes. The organism generally did not persist beyond PE week 2 in other lymph nodes of the head and neck, lymph nodes of the abdominal wall, thoracic cavity, or limbs, or in heart, liver, or spleen. The gallbladder, kidney, and lungs of all pigs were culture-negative.

The efficacy of ivermectin as an in-feed formulation was evaluated against naturally acquired gastrointestinal helmiths, lungworms, and sarcoptic mites (experiment 1; n = 24) and against induced infection with intestinal nematodes (experiment 2; n = 24) in pigs. Treatments consisted of ivermectin administered in feed at concentrations calculated to provide 100 or 200 microgram/kg of body weight/d for 7 days or of nonmedicated feed (controls) for 7 days. At concentration of 100 microgram of ivermectin/kg/d, efficacy against naturally acquired infections was 97.7% for Ascaris suum, 97.8% for Metastrongylus spp, greater than 99% for Oesophagostomum spp, 100% for Macracanthorhynchus hirudinaceus, and 89.7% for Ascarops strongylina. Against induced infections (fourth-stage larvae), efficacy was 100% for A suum and 96.9% for Oesophagostomum spp. At concentration of 200 microgram of ivermectin/kg/d, efficacy against naturally acquired infections was 100% for A suum, Hyostrongylus rubidus, Metastrongylus spp; and 85.9% for Macracanthorhynchus hirudinaceus. Against induced infection (fourth-stage larvae), efficacy was 100% for A suum and 95% for Oesophagostomum spp. At concentrations of 100 and 200 microgram of ivermectin/kg/d, efficacy against Sarcoptes scabiei var suis was evidenced by elimination of the mite by posttreatment day 14.

Immunogenic or pathogenic factors of recombinant proteins (rBCSP20, rBCSP-31, and rBCSP45 of Brucella abortus strain 19) for mice were compared with factors of a proteinase K-treated lipopolysaccharide extracted from B abortus strain 2308. Mice were vaccinated with 4 products, using different inoculation schedules and were challenge exposed with a virulent culture of B abortus strain 2308. Blood samples were collected 2 weeks after vaccination and at necrospy and sera were obtained. Spleens were cultured for B abortus at necropsy (3 to 4 weeks after challenge exposure). Mice given proteinase K-treated lipopolysaccharide alone or in conjunction with rBCSP20 or rBCSP45 proteins were protected, but mice given rBCSP31 on the same day as challenge exposure were not. Vaccination with recombinant proteins alone neither provide protection nor significantly (P greater than 0.05) increase the pathogenic effect of the challenge-exposure culture. Seemingly, rBCSP31 might be a virulence factor of B abortus.

The use of lipoarabinomannan (LAM; obtained from Mycobacterium paratuberculosis) in an ELISA (LAM-ELISA) to test 75 sheep sera from a paratuberculosis-infected flock resulted in an approximate threefold increase in sensitivity (from 23.5% to 70.6%), compared with the use of Annau's polysaccharide in a complement fixation test (P-CFT). Even after manipulation of the LAM-ELISA cut-off value to produce a specificity of 100% to match that of the P-CFT, the sensitivity still was approximately twofold greater than that of the P-CFT. Anti-bovine monoclonal antiglobulin-enzyme conjugates matched commercially available anti-ovine polyclonal antiglobulin-enzyme conjugates with respect to sensitivity and specificity. False-positive results were found to be less frequent after combining 2 serodiagnostic tests, LAM-ELISA and D antigenagar gel immunodiffusion, resulting in an increase in specificity from 88.1% to 95.2%. The repeatability of true seropositive and seronegative results was found to be 89.5% and 91.1%, respectively, for sera obtained less than or equal to 1 month prior to slaughter and 91.7% and 95.5%, respectively, for reanalysis of sera obtained at the time of slaughter.