Objective-To estimate sensitivity and specificity of 4 commonly used brucellosis screening tests in cattle and domestic water buffalo of Trinidad, and to compare test parameter estimates between cattle and water buffalo. Animals-391 cattle and 381 water buffalo. Procedure-4 Brucella-infected herds (2 cattle and 2 water buffalo) and 4 herds (2 of each species) considered to be brucellosis-free were selected. A minimum of 100 animals, or all animals > 1 year of age, were tested from each herd. Serum samples were evaluated for Brucella-specific antibodies by use of standard plate agglutination test (SPAT), card test (CT), buffered plate agglutination test (BPAT), and standard tube agglutination test (STAT). A Bayesian approach was used to estimate sensitivity and specificity of diagnostic tests without the use of a gold standard, assuming conditional independence of tests. Results-Sensitivity and specificity estimates in cattle, respectively, were SPAT, 66.7 and 98.9; CT, 72.7 and 99.6; BPAT, 88.1 and 98.1; and STAT, 80.2 and 99.3. Corresponding test estimates in water buffalo, respectively, were SPAT, 51.4 and 99.3; CT, 90.4 and 99.4; BPAT, 96.3 and 90.7; and STAT, 75.0 and 98.8. Sensitivity of the CT and specificity of the BPAT were different between cattle and water buffalo with at least 95% probability. Conclusions and Clinical Relevance-Brucellosis serologic test performance varied by species tested, but BPAT had the highest sensitivity for screening cattle and water buffalo. Sensitivity and specificity of more than 2 screening tests can be estimated simultaneously without a gold standard by use of Bayesian techniques.

Objective-To use antibodies produced by calves in response to infection with Mannheimia haemolytica in immunoaffinity chromatography for the identification and subsequent isolation of the dominant immunogenic antigens from bacteria grown in irondeficient media. Sample Population-Serum from 10 calves actively infected with M haemolytica. Procedure-An outer membrane protein fraction was obtained from sonicated salt-extracted M haemolytica cells by extraction with N-lauroyl sarcosinate. The immunoglobulin fraction of serum from calves actively infected with M haemolytica was used to prepare an immunoaffinity column. The immunoaffinity column was used to isolate the dominant immunogenic proteins from the outer membrane protein fraction. The resultant immunogenic protein fraction was subjected to ELISA and immunoblot methods as well as carbohydrate quantification. Sequencing of the N-terminal was performed on the most prominent protein. Results-5 immunogenic proteins with molecular weights of 42, 30, 24, 20, and 15 kd were isolated. The immunogenic protein fraction was found to contain 51% carbohydrate. The immunoaffinity column capacity was 1 µg of immunogenic protein/mL of gel. The N-terminal sequence of the 42-kd protein was Tyr-Gln-Thr-Tyr-Gln-Ser-X-Leu-Gln, where X could not be identified. Conclusions and Clinical Relevance-Immunogenic proteins were isolated by use of immunoaffinity chromatography. A substantial amount of carbohydrates was co-purified in the process. Additional experiments are needed to determine whether the carbohydrates would hinder or enhance development of vaccine preparations. This method could potentially allow a more rapid production of antigens for use in vaccines.

Objective-To provide an epidemiologic investigation of the seroprevalence of antibodies to Toxoplasma gondii in populations of cats and wild rodents in Rhode Island and to address the possible epidemiologic role of wild rodents in the spread of toxoplasmosis. Animals-200 cats and 756 small wild rodents. Procedure-Serum samples were obtained from 84 cats in animal shelters and 116 cats in veterinary hospitals. Serum samples were also obtained from 756 small wild rodents from multiple sites in Rhode Island. Sera from rodents and cats were assayed for antibodies to T gondii by use of the modified agglutination test Results-Overall, 42% (84/200) of cats had serum antibodies to T gondii. Seroprevalence was not significantly different between stray (50%; 42 /84) versus client-owned (36%; 42/116) cats, between male (43%; 40/94) versus female (42%; 39/93) cats, or between indoor (26%; 7/27) versus outdoor (39%; 35/89) cats. Seroprevalence rate of trapped rodents was 0.8% (6/756). Six rodents captured in Washington County accounted for of the seropositive rodents. Four of 6 of the seropositive rodents were trapped at a single site in Washington County (an abandoned barn). Five stray cats, known to have resided at the same site in Washington County as 4 of the seropositive rodents, were also found to be seropositive for antibodies to T gondii. Conclusions and Clinical Relevance-Seroprevalence rate in rodents was not correlated with the seroprevalence rate in cats. Stray cats, especially those known to be feral, may be more likely to perpetuate the cat-mouse cycle of T gondii than clientowned cats.