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PubMed1976

Vaccination against bovine brucellosis with a low dose of strain 19 administered by the conjunctival route.

Eighteen 25 month old cows were vaccinated once either with 10(6) to 10(10) living bacteria of Brucella abortus B 19 by the conjunctival route, or with 2.5 to 5 X 10(10) formalin-killed bacteria of the same strain by subcutaneous injection. Fifteen days post-vaccination, low but definite agglutination titers were present in sera of cattle receiving 10(10) living bacteria by the conjunctival route, whereas high titers were observed in the control group given subcutaneous injection of killed bacteria (table I). Agglutination reactions were negative in sera from all cattle receiving less than 10(10) bacteria, and complement fixation tests were negative with all sera. Seventy-one 4 month old claves were vaccinated by the conjunctival route with different doses (10(6) to 10(10)) of living bacteria, either freshly prepared or lyophilized. At autopsy about 15 or about 30 days later, the parotid, submaxillary and retropharyngeal lymph nodes were removed and cultured on Brucella selective medium. Blood samples were taken at autopsy for testing by agglutination, complement fixation, Coombs test and (for some samples) by the Rose Bengal plate test. Colonization of at least one lymph node was observed in 26 of 34 calves given 10(9) bacteria, in all 9 calves given 5 X 10(9) bacteria and in all 12 calves given 10(10) bacteria. Serological responses were demonstrated rarely, and then only in agglutination and Coombs tests. The 5 X 10(9) dose, wich following conjunctival inoculation consistently colonized the regional lymph nodes without inducing significant serological response, would be the most suitable dose for the vaccination of calves and cows.

Animals↗
PubMed1978

In vitro lymphocyte transformation as a herd survey method for bovine paratuberculosis.

The lymphocyte-transformation (LT) test was evaluated for its potential application as a field test for bovine paratuberculosis. Using a whole blood technique, samples from 3 consecutive collection periods were subjected to 3 mycobacterial antigens and to phytohemagglutinin. The results obtained from LT were compared with conventional serologic and cultural methods. A positive LT response to johnin purified-protein derivative (PPD) or avian PPD (or both) was noted in 40% to 60% of the animals tested. The complement-fixation test yielded 4% to 6.7% positive results, the immunodiffusion test between 1.2% and 1.4%, and the direct fecal culture between 2.4% and 6%. The mean of the stimulation indices of all positively responding animals was highest with johnin PPD. Specific stimulation to mammalian PPD occurred between 2.4% and 6% of the animals. The efficacy of the LT test for determining the incidence of infection with Mycobacterium paratuberculosis is discussed.

Animals↗
PubMed1975

[The closed hexagon immunodiffusion (CHI) for the diagnosis of onchocerciasis (author's transl)].

1273 sera from microfilaria-positive and -negative persons living in an onchocerciasis-endemic area in Liberia were examined. For this purpose a modification of the Ouchterlony-test was performed and compared with other methods. In the complement-fixation test many sera showed complement inhibition without any possibility to eliminate it. There were considerably varying titers in the indirect hemagglutination test. This helped to distinguish groups with different correlations to microfilaria-findings. The closed hexagon immunodiffusion after TRAAVIK, SIEBKE and KJELDSBERG (1972) - a modified OUCHTERLONY-test - proved to be nearly as good as indirect hemagglutination. An advantage of the CHI test is that it is easy to carry out. CHI reacted in 64% of microfilaria-positive and in 25% of microfilaria-negative cases.

Complement Fixation Tests↗
PubMed2006

Protective effect of the Nramp1 BB genotype against Brucella abortus in the water buffalo (Bubalus bubalis).

We tested 413 water buffalo cows (142 cases and 271 controls) for the presence of anti-Brucella abortus antibodies (by the skin test, the agglutination test, and the complement fixation test) and the Nramp1 genotype (by capillary electrophoresis). Four alleles (Nramp1A, -B, -C, and -D) were detected in the 3' untranslated region of the Nramp1 gene. The BB genotype was represented among only controls, providing evidence that this genotype confers resistance to Brucella abortus. The monocytes from the BB (resistant) subjects displayed a higher basal level of Nramp1 mRNA and a lower number of viable intracellular bacteria than did the monocytes from AA (susceptible) subjects. The higher basal level of the antibacterial protein Nramp1 most probably provides the BB animals with the possibility of controlling bacteria immediately after their entry inside the cell.

3' Untranslated Regions↗
PubMed2002

Antibody dynamics in Holstein Friesian heifers vaccinated with Brucella abortus strain 19, using seven serological tests.

The serological response induced by Brucella abortus strain 19 was evaluated in 52 Holstein females from a brucellosis-free herd using seven serological tests. Each calf was vaccinated at an age of 4 and 8 months old with 3 x 10(10) CFU B. abortus S19 and the antibody response was determined as the proportion of positive results to each test. The antibody dynamics, measured with the buffered plate antigen (BPA) test and the rapid automated presumptive (RAP) test, were similar. The proportion of positive reactions in these tests reached 100% one week after vaccination and remained at this level for seven weeks, after which the proportion of positive samples slowly declined to 8% (BPA) and 2% (RAP) at week 50. The response in the indirect enzyme immunoassay (i-ELISA) was similar, but shorter than that observed with the BPA/RAP. The antibody dynamic, measured using the seroagglutination test (SAT) in parallel with the 2-mercaptoethanol (2-Me) test and the complement fixation test (CFT) were similar to the RAP/BPA, but of slightly shorter duration. The competitive ELISA (c-ELISA) was positive in all animals for 3 weeks, followed by a rapid decline. The fluorescence polarization assay (FPA) reached a maximum of 68.5% positive animals at week 4 and then declined. Based on these data, the c-ELISA and FPA discriminated residual antibody activity due to vaccination more efficiently than the other tests.

Animals↗
PubMed1978

Post-mortem diagnosis of chronic Chagas's disease comparative evaluation of three serological tests on pericardial fluid.

In an attempt to improve the post-mortem diagnosis of Chagas's disease the authors performed haemagglutination tests (HAT), fluorescent Trypanosoma cruzi antibody tests (FAT), and complement fixation tests (CFT) on the pericardial fluid obtained at autopsy of 50 individuals with Chagas's heart disease, and 93 patients in whom this disease was not thought to be present. The results demonstrate that all three tests are efficient for the post-mortem diagnosis of Chagas's disease but suggest that their combined use would detect more cases than would one isolated reaction only.

Antibodies↗
PubMed1987

Comparison of competitive and indirect enzyme-linked immunosorbent assays for detection of bluetongue virus antibodies in serum and whole blood.

An indirect (I) enzyme-linked immunosorbent assay (ELISA) and a competitive (C) ELISA, using a group-specific monoclonal antibody against bluetongue virus (BTV), are described for the detection of antibodies to BTV in cattle and sheep sera. The performance of these assays in detecting anti-BTV antibody in sequential serum samples and eluates from whole blood (WB) dried on filter paper from three calves and four sheep experimentally infected with type 10 BTV was evaluated. The C-ELISA was superior to the I-ELISA in the detection of anti-BTV antibody in the sera and WB samples from both cattle and sheep early after infection with BTV. BTV antibodies were demonstrable by C-ELISA in all the bovine and ovine sera and WB eluates by 9 days postinfection; whereas the I-ELISA results for sheep sera and WB eluates were similar, anti-BTV antibody was not detected in bovine serum and WB eluates until 26 and 14 days postinfection, respectively. While both ELISAs proved reliable, under the present test conditions involving detection of early postinfection reactions of experimentally infected animals, the C-ELISA was always as sensitive or more sensitive than the standard agar gel immunodiffusion test, the modified complement fixation test, and the plaque neutralization tests in the detection of anti-BTV antibodies. Unlike observations with the immunodiffusion test, no reaction was seen between BTV antigen and bovine epizootic hemorrhagic disease virus antiserum in either ELISA. The results suggest that either ELISA may be suitable for routine diagnostic testing and may have the potential to replace other tests for detection of anti-BTV group-specific antibodies and that the C-ELISA may have the most potential.

Animals↗
PubMed1987

Monitoring of dairy herds for Brucella abortus infection when prevalence is low.

A total of 2,698 dairy herds were surveyed in 1981-1982 in New South Wales and north eastern Victoria in a review of the methods used to monitor them for the presence of Brucella abortus. The methods used to monitor dairy herds were testing of all breeding cows over 1 year of age using the rose bengal test (RBT) and complement fixation test (CFT), the bulk milk ring test (BMRT), and testing of blood samples collected at abattoirs using the RBT and CFT. The surveyed herds had at least one whole herd test, and BMRT was done at regular intervals in the period of the survey. Of the 99 (3.7%) herds that reacted to the BMRT, 91 (3.4%) herds had false positive reactions and 8 (0.3%) herds were declared infected on follow-up herd testing. False-positive reactions were obtained in 22 herds on more than one occasion. Common causes of false positive reactions to the BMRT were thought to be previous vaccination with Strain 19 and sampling in very early or late lactation. Of the 98 (3.63%) herds that reacted to the whole herd serological tests, 80 (2.96%) herds had false-positive reactions and 18 (0.67%) herds were declared infected. Strain 19 vaccination was thought to be an important cause of false-positive reactions. Fifty-three (2.0%) herds showed suspicious reactions on abattoir monitoring but none was declared infected on follow-up testing. Of the 18 herds with infected or equivocal status, the BMRT identified?(ABSTRACT TRUNCATED AT 250 WORDS)

Abattoirs↗
PubMed1970

The passive haemagglutination test for th detection of Mycoplasma suipneumoniae and the possible diagnosis of enzootic pneumonia of pigs.

Fourteen cases of enzootic pneumonia, nearly all of which had presented diagnostic difficulties using the metabolic-inhibition test, were re-examined using specific pig antisera in the passive haemagglutination test (PHA). All proved positive for Mycoplasma suipneumoniae, indicating that the test, used in this manner, might be particularly valuable for routine diagnosis.The PHA test was also used to demonstrate antibody to M. suipneumoniae in pneumonic tissue and the associated bronchial lymph nodes.To allay our concern that cross-reactions might interfere with this and other serological tests-the complement-fixation test (CF) and precipitation in agar-gel-the specificity of our reagents and the antigenic relationships of Mycoplasma hyorhinis, Mycoplasma granularum, mycoplasma B3, Mycoplasma hyopneumoniae and three strains of M. suipneumoniae (including cloned and uncloned isolates of the J strain) were studied in various ways. Antibodies to medium constituents occurred in rabbit antiserum but did not present a problem with pig antisera. These antibodies were successfully absorbed from the rabbit antisera but it was not possible to remove medium constituents from the antigens used to produce antisera in rabbits by repeated washing.By all these tests, the main species of mycoplasmas studied seemed to be antigenically distinct. No major antigenic differences between the three strains of M. suipneumoniae were revealed by the PHA test and the CF test; a slight difference in the precipitation lines of one of these strains (MG) in agar-gel might have indicated an antigenic variation or been a measure of some other factor.

Animals↗
PubMed1983

Applied serology in the latter stages of the eradication of bovine brucellosis.

Late in the program to eradicate bovine brucellosis from Western Australia, Rose Bengal test (RBT) and complement fixation test (CFT) results on the serums from 2,307 cattle (from herds where infection was still present after a minimum of 3 complete herd tests) showed that 327 were positive in the CFT and 246 were positive in the RBT (p less than 0.001). Subsequent testing by the RBT, CFT and the indirect haemolysis test (IHLT) of 722 serums from cattle slaughtered as part of infected herds showed that of 177 cattle positive on culture, 138 were positive in all 3 tests, 9 were negative in all 3 tests and no animal positive on culture had a reaction only in the RBT. In the 177 cattle from which B. abortus was isolated, positive reactions in the CFT occurred in the serums of 159 of them. Application of the RBT as a screening test followed by a confirmatory CFT would have resulted in 149 of the 177 cattle being positive and application of the CFT/IHLT (double test) on the serums of all cattle in the herds would have resulted in 168 or the 177 being regarded as positive.

Animals↗
PubMedDate not supplied

[An enzyme-linked immunosorbent assay (ELISA) for the detection of anti-chlamydial antibodies in pig sera].

The evaluation of an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against chlamydiae in pig sera is described. The most widely used serological test is the complement fixation test (CFT). The CFT has a lack of sensitivity and specificity because of low antibody titers and unspecific reactions. Eight conventionally raised pigs were exposed to a pathogenic strain of Chlamydia suis, four controls were mock infected. The immune responses was monitored by CFT and indirect ELISA. There was no agreement between CFT and ELISA data. These results were confirmed by a study with 191 sera from nine pig farms. As shown by ELISA and PCR chlamydiae are widespread in swine.

Animals↗
PubMed1994

Efficacy of different Rose Bengal and complement fixation antigens for the diagnosis of Brucella melitensis infection in sheep and goats.

Fifteen different Rose Bengal antigens showed large differences with respect to pH, cell concentration and agglutination with the international standard anti-Brucella abortus serum, demonstrating the lack of international standardisation. Their sensitivity and specificity, compared with that of the complement fixation test, were evaluated for the diagnosis of B melitensis infection in culture-positive sheep, brucella-free ewes, and sheep and goats belonging to field flocks under different epidemiological conditions. All the Rose Bengal antigens and the complement fixation test had 100 per cent specificity when testing brucella-free sheep or animals belonging to flocks in unvaccinated brucellosis-free areas, but there were large differences in sensitivity between the Rose Bengal antigens with sera from culture-positive sheep or from animals belonging to infected flocks. When using the most sensitive antigen, no difference was observed in Rose Bengal sensitivity between animals infected with either biovar 1 or biovar 3 of B melitensis. The relationship between the sensitivity of the Rose Bengal antigens and cell concentration was unclear, but their sensitivity was related to the standardisation of the antigens with the international standard serum. The complement fixation test was less sensitive than the Rose Bengal test when testing culture-positive sheep. When testing sera from animals belonging to infected flocks with antigens standardised according to European Union rules, no great differences were observed in the sensitivities of the two tests. However, great differences in sensitivity between the Rose Bengal antigens were observed with sera from animals belonging to flocks with low levels of prevalence.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2001

Serodiagnosis of human cysticercosis by using antigens from vesicular fluid of Taenia crassiceps cysticerci.

Neurocysticercosis (NC), caused by the presence of Taenia solium metacestodes in tissues, is a severe parasitic infection of the central nervous system with universal distribution. To determine the efficiency of enzyme-linked immunosorbent assay (ELISA) and immunoblot with antigens of T. crassiceps vesicular fluid (Tcra) compared to standard techniques (indirect immunofluorescence test [IFT] and complement fixation test [CFT]) using T. solium cysticerci (Tso) for the serodiagnosis of NC, we studied serum samples from 24 patients with NC, 30 supposedly healthy individuals, 76 blood bank donors, 45 individuals with other non-NC parasitoses, and 97 samples from individuals screened for cysticercosis serology (SC). The sensitivity observed was 100% for ELISA-Tso and ELISA-Tcra, 91.7% for the IFT, and 87.5% for the CFT. The specificity was 90% for ELISA-Tso, 96.7% for ELISA-Tcra, 50% for IFT, and 63.3% for CFT. The efficiency was highest for ELISA-Tcra, followed by ELISA-Tso, IFT, and CFT. Of the 23 samples from SC group, which were reactive to ELISA-Tso and/or ELISA-Tcra, only 3 were positive to immunblot-Tcra (specific peptides of 14- and 18-kDa) and to glycoprotein peptides purified from Tcra antigen (gp-Tcra), showing the low predictive value of ELISA for screening. None of the samples from the remaining groups showed specific reactivity in immunoblot-Tcra. These results demonstrate that ELISA-Tcra can be used as a screening method for the serodiagnosis of NC and support the need for specific tests for confirmation of the results. The immunoblot can be used as a confirmatory test both with Tcra and gp-Tcra, with the latter having an advantage in terms of visualization of the results.

Animals↗
PubMed2002

Apparent prevalence of dourine in the Khomas region of Namibia.

A 15-year record of the results of horse sera from the Khomas region of Namibia tested by the complement fixation test for dourine at the Central Veterinary Laboratory in Windhoek before clearing the respective animals for export and competitive sport were subjected to statistical analysis. The range of percentage positive, taken as the apparent prevalence of dourine for the region, during the period of study, was 0-29.09%; the average regional level of apparent prevalence was 8.33%. These figures were thought to be lower than the real situation due to some bias in the sampling criteria. For more accurate results, the more reliable enzyme-linked immunosorbent assay techniques are recommended for use in sero-surveys for dourine in Khomas and other regions of Namibia to provide a basis for development of effective control strategies against the disease.

Animals↗
PubMed1985

Evaluation of three serological tests for detection of antibody to Pseudomonas aeruginosa in human sera.

Four hundred and ninety-five sera from 325 patients from whom Pseudomonas aeruginosa had been isolated and 86 control sera were tested for antibody by indirect haemagglutination tests (HAT) and complement fixation tests (CFT) using a polyvalent pseudomonas serotype-specific vaccine antigen, PEV-02. Sera were also tested by countercurrent immunoelectrophoresis (CIE) for precipitins to a species-specific protein antigen. Control sera gave titres of 160 or less by HAT and 20 or less by CFT. 2-Mercaptoethanol resistant antibody titres (immunoglobulin G) were below 40 for all control sera and none of the latter contained precipitins to common antigen. Of 325 patients, 156 (48%) gave titres of 320 or greater by HAT and of these, 114 (73%) showed elevated immunoglobulin G titres. Less patients with positive blood cultures than expected were positive by HAT and more patients with bone infections gave raised immunoglobulin G titres than expected. Cystic fibrosis patients were invariably seropositive by all tests. There was a correlation between positive CIE and CFT tests, especially in patients who were positive by HAT. Approximately half of 83 patients tested gave a serotype-specific antibody response. The tests were of little value in confirming clinically evident acute infections, but in cases of doubtful infection they did provide confirmatory evidence of an antibody response in approximately one-third of patients culture-positive for Pseudomonas aeruginosa.

Antibodies, Bacterial↗
PubMed1980

A serological survey for bluetongue virus antibody in Western Australia.

A serological survey was carried out to detect specific (serotype 20) and a group bluetongue virus antibody in cattle and sheep serums collected in Western Australia during the period January 1 1978 to June 30 1979. Of 18,849 cattle serums examined by the gel diffusion precipitin test (GDPT), 9.7% were positive and 6.1% gave doubtful results. All 1949 sheep serums tested were negative. Precipitin antibody was demonstrated in 22.5% of serums from Kimberley cattle and 3.6% of cattle serums from the Northwest. Serums collected from cattle in the South were consistently negative in GDPT. When 915 serums that reacted in the GDPT were further tested by the complement fixation test (CFT), 164 were positive. The percentage of CFT positive serums increased as the GDPT reaction became stronger. 2467 serums collected from cattle in Kimberley and Northwest areas and tested by the CFT, 175 (7.1%) were positive. These 175 positive serums were also examined by GDPT and 164 doubtful or positive reactions were obtained. The virus neutralisation (VNT) using serotype 20 virus was carried out on 3804 serums, including all serums that reacted in the GDPT, and 57 were positive. When the VNT positive serums were examined in the other 2 tests, 47 serums were either positive or doubtful in the GDPT and 8 were positive in the CFT. The presence of bluetongue virus group antibody in cattle serums closely followed the suggested distribution pattern of Culicoides brevitarsis but specific serotype 20 neutralising antibody was limited to cattle serums from stations situated north of latitude 17 degrees S in an area of mean annual rainfall higher than 700 mm.

Animals↗