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1.
Lymphocytes from Brucella abortus field strain infected, strain 19 vaccinated, non-exposed and field strain infected, but immunologically unresponsive cattle were incubated with B. abortus antigen and indomethacin. There were significant increases (P < 0.005) in the blastogenic responses, as measured by [3H] thymidine uptake, in cultures with indomethacin as compared to cultures without indomethacin. Lymphocyte blastogenic responses to B. abortus antigen were potentiated by indomethacin in both B. abortus exposed and non-exposed cultures. However, potentiation of sensitized lymphocyte blastogenic responses by indomethacin was significantly greater (P < 0.005) than that in non-exposed lymphocytes. Additionally, indomethacin significantly potentiated Brucella-induced lymphocyte blastogenic responses in lymphocytes from anergic cattle.  相似文献   

2.
New and currently used serological procedures were evaluated using sera from cattle that were challenged with B. abortus S544 (S544) after vaccination with either B. abortus S19 (S19) or B. abortus 45/20 (S45/20) as calves or adults. In animals vaccinated with S19, titres to the indirect haemolysis test (IHLT) rose more slowly, declined more rapidly and involved fewer animals than did titres to the complement fixation test (CFT). In animals vaccinated with S45/20 the rough antigen complement fixation test (RCFT) showed persistent titres. At slaughter the IHLT and CFT were found to be more specific and more sensitive than the Rose Bengal Plate Test (RBPT) and Serum Agglutination Test (SAT) in the detection of cattle infected with B. abortus.  相似文献   

3.
Neonatally thymectomized and normal Holstein-Friesian calves were exposed to bovine viral diarrhea virus and challenged 22 days later. Baseline values in non-specific mitogen induced lymphocyte blastogenesis and serologic responses to tetanus toxoid and Brucella abortus strain 19 were established in neonatally thymectomized and normal calves.Neither viral recovery nor protective antibody production followed the very low level primary exposure to bovine viral diarrhea virus. The expected clinical response to an appropriate challenge inoculum did not occur, however, suggesting an immune response to bovine viral diarrhea virus had been initiated by the primary exposure. Five of eight calves developed protective serum levels of bovine viral diarrhea antibody. This may represent an example of immunologic priming. Exposure to low levels of bovine viral diarrhea virus also has potential implications in the pathogenesis of chronic bovine viral diarrhea wherein low level exposure may interfere with the development of protective levels of bovine viral diarrhea antibody as was observed in three calves.Circulating lymphocyte concentrations and results of non-specific mitogen stimulation of lymphocytes revealed a reduced phytohemagglutinin response in conjunction with a slightly increased response to bacterial lipopolysaccharide and pokeweed mitogen in the presence of moderate lymphopenia. The capacity of lymphocytes to respond to phytohemagglutinin was unchanged but an increased total response, as occurred in control thymectomized and intact calves, did not occur.The response to tetanus toxoid, a thymus-independent antigen, was increased following exposure to bovine viral diarrhea virus, perhaps reflecting an alteration in thymic-independent regulation. The lack of a differential response to Brucella abortus strain 19 implied competence in thymus-dependent helper cells.  相似文献   

4.
Summary

The lymphocyte transformation test (using an in vitro whole‐blood lymphocyte stimulation procedure) and the Brucellin skin test were applied to five heifers infected with virulent Brucella abortus strain 544, five cows inoculated with Yersinia enterocolitica serotype 09, and four non‐exposed cows. Lymphocytes from Brucella‐inoculated animals persistently gave very high blastogenic reactions indicative of active Brucella infection. The test was persistently negative in Yersinia‐infected and non‐exposed cattle. Four of thefive cowsinfected with Yersinia enterocolitica type 09 and allfour control cattle were persistently negative to the delayed hypersensitivity skin reaction with brucellin. All cattle infected with Yersinia enterocolitica type 09 were strongly positive to the Rose Bengal, Serum agglutination, Complement fixation and Antibovine globulin tests using Brucella abortus antigens. One lactating cow infected with Yersinia enterocolitica type 09 was positive to Brucella milk ring test. These results indicate that standard Brucella serological tests are unreliable in differentiating the two infections in cattle and that both the Lymphocyte transformation and brucellin skin tests could be used to differentiate bovine brucellosis from yersiniosis.  相似文献   

5.
The effect of early removal of cows excreting pathogenic Brucella abortus following Strain 19 vaccination of beef cattle herds was determined by comparing cumulative incidence (CI) of brucellosis reactors post-vaccination (p.v.). Adult female cattle in six herds were tested, reactors removed, and vaccinated with 3×109 colony-forming units of B. abortus Strain 19. Cattle were tested at 2 months p.v. and culture-positive cattle were removed from a principal cohort of three herds at approximately 3 months p.v., and removal from a control cohort of three herds delayed until approximately 7 months p.v. Neither CI nor time to eliminate brucellosis was significantly different between cohorts.A review of the parturition data revealed that more of the infected cows in the principal cohort herds terminated gestation before removal. These data suggest that stage of gestation plus diagnostic and management alternatives to prevent parturition of infected cattle in the herd are more important factors in herd plans than early removal of postparturient infected cows following whole-herd Strain 19 vaccination.  相似文献   

6.
Smooth lipopolysaccharides (SLPS) from Brucella abortus contain A-epitopes against which the majority of serum antibodies are directed during infections. SLPS from Yersinia enterocolitica 0:9 possesses identical epitopes, which are the cause for serological cross-reactivity. All Brucella spp. possess M- and C-epitopes which are not present in Y. enterocolitica 0:9. In order to examine the usefulness of these M- and C-epitopes for discrimatory serological testing, a panel of sera were used in this study, comprising sera from Y. enterocolitica 0:9-infected heifers, sera from B. abortus-infected cattle of comparable strength in the serological brucellosis tests to the sera from Y. enterocolitica 0:9-infected heifers, sera from B. abortus-infected bovines with strong serological reactions and sera from animals free from B. abortus or Y. enterocolitica infections. These sera were tested in blocking ELISAs with seven M- and one C-epitope-specific monoclonal antibodies in combination with SLPS from B. melitensis M16 high in M-epitopes as antigen. Strong B. abortus sera inhibited most strongly, while negative sera showed no or little inhibition. Sera with weak or intermediate titres blocked to a lower extent. Unexpectedly, the sera from Y. enterocolitica 0:9-infected heifers showed inhibition behaviour virtually identical to the comparable sera from B. abortus infected animals. Absorbing out of the A-epitope specific serum antibodies with either Y. enterocolitica 0:9 SLPS or with Y. enterocolitica 0:9 bacteria, indicated the presence of M- or C-epitope-specific serum antibodies in some sera from B. abortus-infected cattle but not in the sera from Y. enterocolitica 0:9-infected animals. These results demonstrate that the M- or C-epitope-specific antibody response in sera from B. abortus infected cattle is only of limited value for the serological discrimination between B. abortus and Y. enterocolitica 0:9 infections.  相似文献   

7.
Forty-eight cattle were sensitised toBrucella antigens either by vaccination withBrucella abortus strain 19 (S19) orB. abortus 45/20 (S45/20) and 24 of these and 12 unvaccinated cattle were subsequently challenged with virulentB. abortus strain 544 (S544). All these cattle (n=60), together with 12 control cattle which were neither vaccinated nor challenged, were subsequently subjected to an intradermal test using a S45/20 protein antigen. Reactions were interpreted subjectively by observation and palpation and were measured to the nearest mm with calipers at 48 and 72 hours after injection of protein antigen. Ten weeks later the cattle were slaughtered and tissues cultured for the presence ofB. abortus. Two of the 48 vaccinated cattle died, 40 of the remaining 46 gave a positive response to the intradermal test at 48 hours and 36 were positive at 72 hours. In the controls any increase in the skin thickness had disappeared by 72 hours. An increase in skin thickness was still present at 72 hours in all other cattle except those vaccinated with S19 only. The intradermal test was found to be sensitive but not specific in detecting infected cattle and both sensitive and highly specific if used (with the exception of S19) to detect exposure toBrucella antigen.  相似文献   

8.
White-lipped marmosets were evaluated for their cell mediated immune (CMI) response to EBV to determine the feasibility of CMI studies in marmoset models for EBV oncogenesis. The mitogen, cell concentrations, the length of incubation period and serum requirements were defined for in vitro lymphocyte stimulation tests. The level of response of each animal was dependent on the concentration of phytohemagglutinin-P (PHA-P) and was independent of cell densities employed. The rate of tritiated-thymidine incorporation by mononuclear cells due to PHA-P increased exponentially between 2–4 days. This test was reproducible for a given batch of PHA-P when the cells were cultured in the presence of 10% heat inactivated fetal bovine serum. The five white-lipped marmosets were seronegative for EBV antigens and did not show lymphocyte stimulation with EBV particles and EBV soluble antigen, but two of these animals exhibited significant stimulation with autologous lymphocytes transformed in vitro by B95-8 virus. Despite the limited amount of blood (3–4 ml) that could be obtained from each animal in a single bleeding, it was possible to perform multiple lymphocyte stimulation assays with the protocol used.  相似文献   

9.
Peripheral blood leukocytes separated by erythrocyte aggregation and density-gradient centrifugation were used in tests to measure the cell-mediated response in bovine anaplasmosis. The leukocyte migration inhibition test was apparently related to the host protective immune response. Leukocyte stimulation by a specific anaplasma antigen elicits positive but low level stimulation indices in cultures of cells collected from cattle exposed to live anaplasma. There appears to be a nonspecific enhancement of blastogenesis in cultures of leukocytes from cattle injected with inactivated-oil fortified Anaplasma marginale vaccine. Cytotoxicity tests indicated that both leukocytes and antibodies are active in clearance of anaplasma infected erythrocytes from the circulatory system.  相似文献   

10.
A whole blood lymphocyte stimulation assay utilizing the uptake of tritiated thymidine was developed for the detection of Mycobacterium bovis sensitivity in cattle. Results on eight M. bovis infected animals (six to ten weeks after infection) and eight control animals show that satisfactory lymphocyte stimulation can be obtained using heparinized whole blood diluted 10-fold in tissue culture medium and cultured with purified protein derivative (PPD) for three to seven days. Infected animals exhibited significantly greater stimulation when cultured with PPD than did control animals.  相似文献   

11.
Immunizing animals in the wild against Brucella (B.) abortus is essential to control bovine brucellosis because cattle can get the disease through close contact with infected wildlife. The aim of this experiment was to evaluate the effectiveness of the B. abortus strain RB51 vaccine in protecting infection as well as vertical transmission in Sprague-Dawley (SD) rats against B. abortus biotype 1. Virgin female SD rats (n = 48) two months of age were divided into two groups: one group (n = 24) received RB51 vaccine intraperitoneally with 3 × 1010 colony forming units (CFU) and the other group (n = 24) was used as non-vaccinated control. Non-vaccinated and RB51-vaccinated rats were challenged with 1.5 × 109 CFU of virulent B. abortus biotype 1 six weeks after vaccination. Three weeks after challenge, all rats were bred. Verification of RB51-vaccine induced protection in SD rats was determined by bacteriological, serological and molecular screening of maternal and fetal tissues at necropsy. The RB51 vaccine elicited 81.25% protection in SD rats against infection with B. abortus biotype 1. Offspring from rats vaccinated with RB51 had a decreased (p < 0.05) prevalence of vertical transmission of B. abortus biotype 1 compared to the offspring from non-vaccinated rats (20.23% and 87.50%, respectively). This is the first report of RB51 vaccination efficacy against the vertical transmission of B. abortus in the SD rat model.  相似文献   

12.
One group of 51 cattle was vaccinated with B. abortus S19 (S19) and a further 51 cattle were vaccinated with B. abortus S45/20 (S45/20). Forty-eight cattle (24 from each group) and a control group of 12 cattle were subsequently challenged with B. abortus S544. The enzyme-linked immunosorbent assay (ELISA) was used to detect specific IgG and IgM antibodies in these groups. All cattle vaccinated with S19 had high levels of IgG and IgM, but the S45/20 vaccine produced detectable antibody in only a few cattle. In those cattle where the challenge induced infection, the mean levels of IgG and IgM were much higher than those of the uninfected cattle in the same groups. When the isolation of B. abortus was compared at slaughter with the serological results, the ELISA, when used to detect specific IgG, was more sensitive but less specific than the serum agglutination test, complement fixation test and indirect haemolysis test, and more sensitive and more specific than the Rose Bengal test.  相似文献   

13.
Three dogs reared on a dairy farm with a high incidence for Brucella abortus were serologically positive for B. abortus and no other Brucella spp. The identity of the organism was confirmed to be B. abortus by AMOS (abortus melitensis ovis suis)-polymerase chain reaction with specific primers for B. canis. One hundred percent homology of the canine isolate and the bovine pathogen isolated from the farm was demonstrated. The only possible source of infection was infected cattle on the same farm. It is suggested that dogs be routinely included in brucellosis surveillance and eradication programs.  相似文献   

14.
《Veterinary microbiology》1997,57(4):361-371
The course of immunological reaction in 10 Yersinia enterocolitica 0:9 experimentally-infected heifers was followed using the conventional brucellosis tests complement fixation test (CFT), serum agglutination test (SAT) and brucella card test (BCT), and a recently developed Brucella antigen-specific gamma interferon (IFN-γ) test. Initially, the animals were exposed orally to 1010 colony-forming units (CFU) of Y. enterocolitica 0:9. Four weeks later, they were inoculated intravenously with 108 CFU of Y. enterocolitica 0:9 cells. After oral inoculation, the response in the conventional brucellosis tests was minimal. Only after intravenous inoculation were CFT and SAT titres and BCT reactions comparable to natural, false positive brucellosis reactors. After oral exposure the Brucellergen-stimulated release of IFN-γ peaked at values above the cut-off stimulation index of 2.5 in 80% of the heifers. After intravenous inoculation, stimulation indices above 2.5 were present in only 10% of the animals. Two B. abortus infected control cattle showed stimulation indices of 3.1 and 3.4, and a negative control animal exhibited a stimulation index of 1.0. These findings show, in contrast to a previous study, that the Brucellergen-specific IFN-γ assay cannot be used as a specific and discriminatory test for B. abortus infections.  相似文献   

15.
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16.
Ten strains of Yersinia enterocolitica belonging to ten various serogroups isolated from raw milk were inoculated into groups of five guinea pigs and five calves. Y. enterocolitica serotype 0:16 was the only serotype tested that induced an antibody response to Brucella abortus in calves. No anti-Brucella response could be demonstrated serologically in guinea pigs. Activity of the anti-Y. enterocolitica 0:16 calf sera against B. abortus antigen was shown by the tube agglutination test, and by the complement fixation test. The early agglutinating antibody response was partly sensitive to reduction by 2-mercaptoethanol. This sensitivity decreased later in the response. This is the first report of anti-Brucella responses induced by a serotype of Y. enterocolitica other than 0:9; sera from a group of five calves inoculated with 0:9 were tested by the same serological techniques for comparison.  相似文献   

17.
An outbreak of brucellosis caused by Brucella abortus biovar 2 was identified in cattle in Alberta in December 1986. This was the only clinical infection discovered since the national cattle herd was declared brucellosisfree in 1985. It was the first report of B. abortus biovar 2 in Canadian cattle. The outbreak, involving three herds containing purebred Hereford cattle, was spread by the private treaty sale of untested cattle, and was identified following investigation of an abortion. The source of infection for the outbreak was not established, but several possibilities were identified including infected herds present in the area during the mid-1970's, latent infection originating in a Saskatchewan herd during the early 1960's, American cattle imported during the early 1970's, and brucellosis-infected bison in Wood Buffalo National Park. The containment and elimination of this nidus of infection appears to have been successful, and the national cattle herd at the time of writing is free of the disease.  相似文献   

18.
SUMMARY A group of 4 cows was vaccinated with Brucella abortus strain 19, followed 8 weeks later by a single dose of B. abortus 45/20 vaccine. A similar group received 2 doses of B. abortus 45/20 vaccine 8 weeks apart. The antibody responses of the groups were compared by testing whole serums and separated IgM and IgG fractions by the Rose Bengal Plate (RBP) agglutination and the complement fixation tests (CFT) using rough and smooth B. abortus antigens. Animals that had received B. abortus strain 19 responded to the 45/20 vaccine with increased titres to the smooth antigen. These relevant antibodies were predominantly of the IgG class. Standard CFT and RBP test antibodies could be detected in IgM and IgG fractions after the primary inoculation with B. abortus strain 19 vaccine.  相似文献   

19.
A Brucella abortus-soluble antigen was investigated, using in vitro assay of lymphocyte immunostimulation, to determine which concentration of this antigen and which period of incubation of the lymphocyte cultures would induce maximum specific lymphocyte immunostimulation as an additional method for further study of B abortus infection in cattle. Soluble antigen was prepared from autoclaved cells of B abortus strain 1119-3. Peripheral blood lymphocytes were obtained from cattle infected with B abortus and from healthy control cattle not infected with B abortus. The lymphocytes were prepared by the Ficoll-Hypaque density gradient technique, suspended in RPMI 1640 medium (1.5 X 10(6)/ml), cultured with several dilutions of soluble antigen, and incubated. Prior to termination of incubation, cultures were labeled with 1 muCi of [3H]thymidine and, after harvesting, assayed for [3H]thymidine incorporation in DNA by a liquid scintillation spectrometer. Maximum specific immunostimulation of lymphocytes from B abortus-infected cattle was induced in this assay system with 6 days' incubation and 22 microgram of protein/ml/1.5 X 10(6) lymphocytes, using protein content to express concentration of soluble antigen in this system.  相似文献   

20.
Recent research has shown that cattle that develop mucosal disease (MD) often, if not always, have been persistently infected with bovine viral diarrhea virus (BVDV) since birth. The purpose of the present study was to determine whether MD could be induced by immunosuppression of persistently BVDV-infected cattle. For that purpose, adrenocorticotropic hormone (ACTH) was injected intramuscularly, twice daily for 5 consecutive days in 4 persistently BVDV-infected cattle and in 3 control cattle. Before the ACTH treatment, the numbers of leukocytes, neutrophils and mononuclear cells (MNC) per litre of blood in BVDV-infected cattle were in the same range as in the controls. Similarly, the proportions of B cells, T cells, monocytes and Fcγ+ cells (cells with receptor for the Fc part of IgG) were the same in the 2 groups of animals. On the other hand, the proliferative response to mitogen stimulation of MNC obtained from the control animals was twice as high as the corresponding value of the persistently BVDV-infected cattle.In all animals, ACTH treatment caused increased Cortisol concentrations, leukocytosis, neutrophilia and decreased mitogen-induced lymphocyte stimulation. However, the MNC count and the proportions of B cells, T cells, Fcγ+ cells and monocytes remained unaltered. In spite of the immunosuppression, indicated by the decrease in mitogen-induced lymphocyte stimulation. ACTH treatment did not provoke any clinical signs of MD in the persistently BVDV-infected cattle.  相似文献   

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