An enzyme-linked immunosorbent assay (ELISA) was developed to detect antigen-specific secretory IgA antibody in bovine vaginal mucus after abortion due to Campylobacter fetus subsp venerealis. Abortions were diagnosed by isolating the organism from 8 foetuses and/or foetal membranes and by histopathology. Vaginal mucus was collected from 7 cows shortly after abortion. All showed a high level of IgA antibody in their vaginal mucus when they were compared with an uninfected control group. The new ELISA is simple and practical and provides a useful tool for diagnosis of bovine venereal campylobacteriosis.

Adherence of Tritrichomonas foetus to bovine vaginal epithelial cells (VECs) in vitro was investigated with fresh washed bovine VECs and log-phase cultures of T. foetus. Observation under phase-contrast microscopy showed that T. foetus usually adhered first by the posterior flagellum and later by the body. Significantly more keratinized squamous epithelial cells were detected with attached parasites than nonkeratinized round epithelial cells. The optimal pH range for attachment was 6.0 to 7.5, with peak attachment at pH 6.5 for squamous VECs. Surface-reactive bovine antiserum to T. foetus prevented adherence to bovine squamous VECs. Inhibition of adherence occurred at nonagglutinating, nonimmobilizing serum dilutions. Antiserum fractions enriched for immunoglobulin Gl inhibited adherence, but fractions enriched for immunoglobulin G2 did not. The inhibitory antiserum was specific for several mediumto high-molecular-weight membrane antigens as detected in Western blots (immunoblots). The ability of surface-reactive antibodies to prevent adherence and to agglutinate and immobilize T. foetus indicates that they may be protective.

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Morphology, biochemical reactions, pigmentation, antigens, and cell envelope proteins were examined in 12 strains of Haemophiliis somnuts, Haemophilus agni, Histophiliis ovis, and Actinobacillus seminis. All of the strains except A. seminis are related and are considered as a single Haemophilius-Histophilius (HH) group. In immunodiffusion tests, HH group bacteria had at least two antigens common to all members of the group, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that they have similar cell envelope protein profiles. A quantitatively variable yellow pigment with absorption maxima of 430 to 435 nm was present in strains of H. somnnus and H. agni. The HH group did not produce catalase and grew only in air containing 10% CO2. Of 10 HH group bacteria, 9 required thiamine monophosphate for growth. A. seminis was distinguished from the HH group by its lack of yellow pigment, production of catalase, growth in air, lack of a thiamine monophosphate requirement, and different cell envelope protein profile. In gel immunodiffusion tests, A. seminis antigens produced two lines of partial identity with the HH group when antiserum against H. somnnls was used. Reference strains of Haemophilus inflimenzae, Actinobac(illums lignieresii, and Haemophillis haemoglobinophiliis were compared with the test strains. In immunodiffusion tests, a single antigen was found to be common to H. haemoglobinophiluis, A. seminis, and the HH group. No similarities between any of the test strains and H. infliuenzae or A. lignieresii were noted. The close relationship of H. somnus, H. agni, and Histophillus ovis suggests that these unofficially named bacteria may belong to a single taxon.

Incorporation of vancomycin (5 micrograms/ml), neomycin (5 micrograms/ml), sodium azide (50 micrograms/ml), nystatin (100 iu/ml) and cyclohexamide (100 micrograms/ml) into 5 per cent horse blood agar results in a selective medium for the primary isolation of Haemophilus somnus from cattle and sheep. Addition of thiamine monophosphate (1 microgram/ml) to the medium enhanced growth of this bacterium. Gram-positive bacteria did not grow on the medium and colonies of many Gram-negative bacteria were eliminated or reduced in numbers and size. Colonies of H somnus were larger on the selective medium than on sheep blood agar but retained typical morphology. Recovery of 18 laboratory strains was 73 to 166 per cent (mean 112) on selective medium compared to sheep blood agar. H somnus was isolated from the vagina of a total of 136 (28.6 per cent) of 476 cows surveyed, 79 (16.6 per cent) on sheep blood agar and 129 (27.1 per cent) on selective medium. The selective agents and thiamine were stable indefinitely as a freeze dried mixture while prepared plates were stable for two weeks.