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Brucellosis (caused by the bacterium Brucella abortus) is a zoonotic disease endemic in wild elk (Cervus canadensis) of the Greater Yellowstone Ecosystem, US. Because livestock and humans working with elk or livestock are at risk, validated tests to detect the B. abortus antibody in elk are needed. Using the κ-statistic, we evaluated the buffered, acidified plate antigen (BAPA) assay for agreement with the results of the four serologic tests (card test [card], complement fixation test [CF], rivanol precipitation plate agglutination test [RIV], standard plate agglutination test [SPT]) that are approved by the US Department of Agriculture for the detection of the B. abortus antibody in elk. From 2006 to 2010, serum samples collected from elk within B. abortus-endemic areas (n = 604) and nonendemic areas (n = 707) and from elk culture-positive for B. abortus (n = 36) were split and blind tested by four elk serum diagnostic laboratories. κ-Values showed a high degree of agreement for the card (0.876), RIV (0.84), and CF (0.774) test pairings and moderate agreement for the SPT (0.578). Sensitivities for the BAPA, card, RIV, CF, and SPT were 0.859, 0.839, 0.899, 1.00, and 0.813, whereas specificities were 0.986, 0.993, 0.986, 0.98, and 0.968, respectively. The positive predictive values and the negative predictive values were calculated for 2.6%, 8.8%, and 16.2% prevalence levels. These findings suggest the BAPA test is a suitable screening test for the B. abortus antibodies in elk.

Thirty-one bison heifers were randomly assigned to receive saline or a single vaccination with 10(10) CFU of Brucella abortus strain RB51. Some vaccinated bison were randomly selected for booster vaccination with RB51 at 11 months after the initial vaccination. Mean antibody responses to RB51 were greater (P < 0.05) in vaccinated bison after initial and booster vaccination than in nonvaccinated bison. The proliferative responses by peripheral blood mononuclear cells (PBMC) from the vaccinated bison were greater (P < 0.05) than those in the nonvaccinated bison at 16 and 24 weeks after the initial vaccination but not after the booster vaccination. The relative gene expression of gamma interferon (IFN-γ) was increased (P < 0.05) in the RB51-vaccinated bison at 8, 16, and 24 weeks after the initial vaccination and at 8 weeks after the booster vaccination. The vaccinated bison had greater (P < 0.05) in vitro production of IFN-γ at all sampling times, greater interleukin-1β (IL-1β) production in various samplings after the initial and booster vaccinations, and greater IL-6 production at one sampling time after the booster vaccination. Between 170 and 180 days of gestation, the bison were intraconjunctivally challenged with approximately 1 × 10(7) CFU of B. abortus strain 2308. The incidences of abortion and infection were greater (P < 0.05) in the nonvaccinated bison after experimental challenge than in the bison receiving either vaccination treatment. Booster-vaccinated, but not single-vaccinated bison, had a reduced (P < 0.05) incidence of infection in fetal tissues and maternal tissues compared to that in the controls. Compared to the nonvaccinated bison, both vaccination treatments lowered the colonization (measured as the CFU/g of tissue) of Brucella organisms in all tissues, except in retropharyngeal and supramammary lymph nodes. Our study suggests that RB51 booster vaccination is an effective vaccination strategy for enhancing herd immunity against brucellosis in bison., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Objective: To estimate the prevalence of Mycobacterium avium subsp paratuberculosis infection among cows on beef operations in the United States., Design: Cross-sectional seroprevalence study. Sample Population-A convenience sample of 380 herds in 21 states., Procedures: Serum samples were obtained from 10,371 cows and tested for antibodies to M avium subsp paratuberculosis with a commercial ELISA. Producers were interviewed to collect data on herd management practices., Results: 30 (7.9%) herds had 1 or more animals for which results of the ELISA were positive; 40 (0.4%) of the individual cow samples yielded positive results. None of the herd management practices studied were found to be associated with whether any animals in the herd would be positive for antibodies to M avium subsp paratuberculosis., Conclusions and Clinical Relevance: Results suggest that the prevalence of antibodies to M avium subsp paratuberculosis among beef cows in the United States is low. Herds with seropositive animals were widely distributed geographically.

The sensitivities and specificities of 17 antibody detection tests for brucellosis in goats were estimated. Tests evaluated included the U.S. Department of Agriculture (USDA) card test with 8% cell concentration (8%Card), USDA rapid automated presumptive test (RAP), Mexican rose bengal plate tests with 8 and 3% cell concentrations (8%RB and 3%RB), French rose bengal plate test with 4.5% cell concentration (4.5%RB), USDA standard plate test (SPT), USDA buffered acidified plate agglutination test (BAPA), USDA and Mexican rivanol tests (URIV and MRIV), USDA standard tube tests with Brucella abortus and Brucella melitensis antigens (SATA and SATM), serum enzyme-linked immunosorbent assay (ELISA), USDA cold-fixation complement fixation tests with B. abortus and B. melitensis antigens (CFA and CFM), USDA and Mexican milk ring tests (UBRT and MBRT), and a milk ELISA. Test sensitivity was evaluated by using two groups of 10 goats experimentally infected with B. melitensis or B. abortus and monitored for 24 weeks. Specificity was evaluated by using 200 brucellosis-free nonvaccinated goats from 10 California herds. The 3%RB was considered a good screening test because of high sensitivity at week 24 postinfection (90%), ease of performance, and low cost. The cold-fixation CFA and CFM had 100% specificity in the field study and were considered appropriate confirmatory tests. The milk ELISA was significantly more sensitive (P < 0.05) than the UBRT and significantly more specific (P < 0.05) than the MBRT. The milk ELISA also had the advantage of objectivity and ease of interpretation.

This study was designed to determine if Brucella abortus strain RB51, which expresses small amounts of the lipopolysaccharide O side chain, would cause positive responses on brucellosis serologic surveillance tests when given to adult cattle that were vaccinated as calves with B. abortus strain 19. Cattle vaccinated as adults with strain RB51 that had been vaccinated as calves with strain 19 (n = 40) had significantly greater antibody titers (P < 0.05) against strain RB51 at 4 and 8 weeks postvaccination in the dot blot assay than did animals (n = 10) not vaccinated with strain RB51. When evaluated using the card or buffered acid plate agglutination presumptive tests, 7 strain RB51 vaccinates tested positive at either 4 or 8 weeks following vaccination as compared with 4 cattle in the control group that were not vaccinated with strain RB51. One strain RB51 vaccinate was scored as suspect on the standard tube agglutination (STA) test at 8 weeks following vaccination. Remaining samples from strain RB51 vaccinates tested negative on the STA, complement fixation (CF), rivanol, and particle concentration fluorescence immunoassay (PCFIA) confirmatory tests. Samples from 2 control cattle were PCFIA positive at time 0; 1 of these animals was CF positive throughout the study. This study suggests that use of strain RB51 in cattle vaccinated with strain 19 as calves will not cause positive responses on confirmatory tests and will not impair brucellosis serologic surveillance efforts.

In this study, we demonstrated by a Cu-Zn superoxide dismutase-specific enzyme-linked immunoassay that cattle that are serologically positive for Brucella abortus have serum immunoglobulin G antibodies to B. abortus Cu-Zn superoxide dismutase. The specificity of the antibody reactivity was confirmed by Western blot (immunoblot) analysis with B. abortus salt-extractable proteins containing native Cu-Zn superoxide dismutase and with recombinant B. abortus Cu-Zn superoxide dismutase. The results represent a first step in the direction of the development of a multiprotein diagnostic reagent for bovine brucellosis.

Serologic responses in the particle concentration fluorescence immunoassay and the card, complement fixation, and tube agglutination tests were measured for 10 weeks after vaccination of cattle with either Brucella abortus 19 or the lipopolysaccharide O-antigen-deficient mutant, strain RB51. The responses of strain 19-vaccinated cattle were positive, whereas those of strain RB51-vaccinated cattle were negative, in all of the tests. These results indicate that cattle vaccinated with strain RB51 fail to produce antibodies that can be detected by conventional serologic tests that are used to diagnose bovine brucellosis.