An ELISA was developed to detect antibodies to the 41-kd flagellin (P41) of Borrelia burgdorferi in serum obtained from cattle. Absorption studies, immunoblot analysis, immunoelectron microscopy, and correlation of results of the P41-ELISA and the P39-ELISA as well as measurement of the antibody to P41 in calves challenge-exposed with Borrelia theileri were used to assess the specificity of the P41-ELISA. Antigens derived from Escherichia coli, Leptospira interrogans serovar hardjo, and B burgdorferi were used for absorption studies and immunoblot analysis. Antibodies to P41 of B burgdorferi cross-reacted with antigens of E coli, but were not cross-reactive with L hardjo. A value 3 SD higher than the mean of the negative-control population of cattle was defined as the minimum value cutoff value) for a positive result by the P41-ELISA. Use of this value for classification of test results reduced the predicted rate of false-positive results attributable to E coli cross-reactivity to 1%. Immunoblot analysis revealed that test-positive serum from cattle reacted mainly with 41-, 39-, 34-, and 31-kd proteins of B burgdorferi, as well as several smaller proteins. Immunoelectron microscopy revealed that serum from cattle that was test-positive by the P41-ELISA bound to the flagellin and outer membrane of B burgdorferi. Results of absorption studies, immunoblot analysis, and immunoelectron microscopy were correlated and indicated that serum from cattle that was test-positive by P41-ELISA had stronger reactivity to B burgdorferi antigens than to antigens of E coli or L hardjo. The concentrations of antibodies measured by P41-ELISA and P39-ELISA testing were highly correlated [R(2)=0.78]. Calves challenge-exposed with B theileri also had test-positive results by the P-41-ELISA as early as 2 weeks after exposure, but serum antibody concentrations decreased to prechallenge-exposure concentrations by 9 weeks after exposure.

During the hunting season of 1992, 322 black bears from Pennsylvania were examined for Toxoplasma gondii- and Trichinella spp-induced infections. Toxoplasma gondii antibodies were found in 79.8% of 322 bears--titer < 1:25 in 65 (20.2%), 1:25 in 18 (5.6%), 1:50 in 11 (34.5%) and 1:500 in 128 (38.7%) bears--by use of the modified agglutination test. Muscle tissues from 89 of these bears were bioassayed for T gondii parasites. Muscles from 64 bears, including heart from 1 bear, and heart alone from another bear, were digested in pepsin, and the digested samples were bioassayed in mice. Toxoplasma gondii was isolated from 5 bears; from the heart of 1, heart and skeletal muscles of 1, and skeletal muscles of 3. The T gondii antibody titers for the 5 bears with detectable T gondii were: greater than or equal to 1:25 in all 5 bears by use of the modified agglutination test; < 1:10 (3 bears, considered Toxoplasma-negative), 1:20 and 1:320 by use of the Sabin-Feldman dye test; < 1:64 (3 bears, considered Toxoplasma-negative), 1:128, 1:512 by use of the indirect hemagglutination test, and < 1:16 (2 bears, considered Toxoplasma-negative), 1:32, 1:64, and 1:512 by use of the latex agglutination test. Toxoplasma gondii was not isolated from feces of 5 cats fed muscles from the remaining 25 bears with T gondii antibody titer < 1:25. Tissue cysts of the 4 T gondii isolates from bears were rendered noninfective by freezing at -13 C. Antibodies against Trichinella spp were found in 6 (1.8%) of 319 bear sera; Trichinella spp larvae were detected in muscle digests of 2 of 63 bears, and in histologic sections of muscles from 3 of 162 bears. Genetic typing indicated that the 2 Trichinella isolates from bears were a sylvatic genotype and were not the species found in domestic pigs.

An agar gel immunodiffusion (AGID) test was used over a 3-year period to examine 1,871 serum samples from sheep representing 5 Mycobacterium paratuberculosis infected flocks and 4 flocks presumed to be uninfected. Of 1,032 sheep, 31 had positive AGID test results (scoring 1 to 5), and 23 of these 31 were ecropsied. Infection with M paratuberculosis was confirmed by 1 or more of the following findings: observation of typical lesions on histologic examination of sections of ileum or ileocecal lymph nodes, observation of clumps of acid-fast bacteria in mucosal smears of ileum, and isolation of the organism from feces or tissue. False-positive results on AGID testing were not found in sheep from flocks known to have exposure to Cotynebacterium pseudotuberculosis. Diarrhea in infected sheep was observed infrequency; chronic, severe weight loss was the most common sign observed. On histologic examination of tissues from 20 infected sheep, 16 (80%) had diffuse lesions of the ileum and 13 (65%) had acid-fast bacteria in areas of ileal inflammation; 4 had discrete granulomas and peripheral lymphocytic infiltrates in the ileum. Sheep with diffuse lesions tended to have higher mean scores on AGID testing and examination for acid-fast bacteria, compared with those from sheep with more discrete lesions. Bacteriologic culture yielded M paratuberculosis from only 3 sheep with paratuberculosis. On the basis of results of this study, we suggest that the nature of the response to infection with M paratuberculosis may influence the results of diagnostic tests for paratuberculosis, and that AGID testing may be useful to identify M paratuberculosis infection in sheep with chronic weight loss and in flock-screening programs.

Indirect immunofluorescent antibody (IFA), latex agglutination (IA), and enzyme immunoassay (EIA) methods were compared for evaluation of the serum antibody responses of dogs experimentally and naturally exposed to spotted fever-group rickettsiae. Selected sera (obtained on days 1, 42, 53, 124, 145, 236, 255, 264, and 292) were examined from three 8-month-old female Beagles inoculated with Rickettsia rickettsii on days 34 and 250 of the study. A second group of dogs comprised three 8-month-old female Beagles inoculated with R montana on days 34 and 102. Subsequently, these dogs were inoculated with R rickettsii on day 250. Serum samples were obtained from the second group of dogs on days 1, 96, 103, 132, 180, 215, 292, and 494. A third group consisted of 21 naturally exposed dogs, from which sequentially obtained serum samples were available, and which had clinical signs compatible with Rocky Mountain spotted fever. Clinical signs of disease in dogs of the third group resolved after treatment with tetracycline (22 mg/kg of body weight, Po, q 8 h) was instituted. At least 2 sequentially obtained serum samples from each dog were tested. In general, the first sample was obtained just prior to treatment and the convalescent serum samples were obtained at weekly or greater intervals thereafter. For correlation and reactivity data, an IFA test for IgG/IgM (using heavy and light chains-specific conjugate) was used as the reference standard for comparison of results with those of the other tests,.

In Peninsular Malaysia, footand-mouth disease (FMD) has been reported since early 1860 which then became sporadic, causing outbreaks every year. Since then, Peninsular Malaysia has become endemic with FMD. The aim of this study is to provide findings of the current FMD occurrence and its serotyping in Peninsular Malaysia. An identification of Foot and Mouth Disease serotype was carried out in Peninsular Malaysia by the Regional Veterinary Laboratory Kota Bharu (RVLKB) only. Epithelial tissue samples were received from 10 states throughout Peninsular Malaysia from 2012 until 2016. Indirect sandwich ELISA was performed using ELISA kit for FMDV antigen detection supplied from the Institute for Animal Health, Pirbright Laboratory. All findings and results in this paper were based on samples received by RVLKB and does not reflect overall cases reported to State DVS or to DVS Malaysia. From the results, 2013 had the highest samples positive for FMDV (35% from 43 samples), followed by 2014 (31% from 80 samples), 2012 (24% from 122 samples), 2015 (21% from 39 samples) and the lowest is 2016 (17% from 194 samples). The FMDV serotypes detected throughout 2012 to 2016 from 110 positive samples were Serotype O (80%), followed by Serotype A (20%) and none from Serotype Asia 1. Strict regulation, FMD vaccine evaluation by LPB ELISA and strict animal movement shall be considered to achieve FMD free for upcoming Year 2020.

A study was conducted to identify the current status of Fasciolaand Paramphistomum infections in small ruminants in Terengganu. A total of 267 faecal samples from small ruminants were collected and subjected to sedimentation technique. Serum samples were diagnosed for detection of IgG antibody for Fasciola infection using sELISA method. Results showed that there were 4% of the goats positive with Paramphistomum eggs whereas Fasciola egg was not observed in any of the faecal samples. However, it was found that 89% of the serum samples from goats were positive with IgG antibody for Fasciola infection. Small ruminants in Terengganu were not infected with severe Fasciola and Paramphistomum infections yet the results obtained from this study will update the current status of the infections. This information will help the farmers and the Department of Veterinary Services to plan on management to maintain the animals’ health.

Recombinant LipL32 and LipL41 outer membrane proteins of Leptospira interrogans serovar Canicola were produced, and used as a pooled antigen in enzyme-linked immunosorbent assay (ELISA) to detect leptospiral antibodies in bovine sera samples. The optimum concentration of the pooled antigen was found to be 50ng of each antigen per well by using known positive and negative cattle sera. Using a total of 500 bovine sera samples the sensitivity, specificity and accuracy of pooled antigen based ELISA as compared to microscopic agglutination test (MAT) were 100%, 88.1% and 91.6%, respectively. The results suggested that antigen in ELISA could be preferred for detection of all those cases, which might have remained undiagnosed by performing MAT.