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1.
本试验采用SAT方法对乌鲁木齐地区某牛场进行了布鲁氏菌病监测,对阳性奶牛的乳汁进行细菌分离培养,用VirB8-PcR方法对分离株VirB8基因进行扩增鉴定。结果表明,2个分离株均为布鲁氏菌,布鲁氏菌分子分型PCR的鉴定结果显示该牛场感染的自然菌株为布鲁氏菌牛种（3b,5,6,9型）。  相似文献   

2.
Brucella suis biotype 1 was isolated from the semen of a dog with hindlimb weakness and a large, firm, left epididymis. A semen sample was oligospermic, with many neutrophils, the numbers of which decreased in serial sampling. A card agglutination test for B abortus and a rapid slide agglutination test for B canis were positive. The modified 2-mercaptoethanol slide agglutination test for B canis and the agar gel immunodiffusion test, using B canis cell wall antigen, were negative. At necropsy, chronic granulomatous inflammation was found in, and B suis biotype 1 was isolated from, the left epididymis and prostate gland.  相似文献   

3.
A double immunodiffusion test for precipitins against Brucella antigen A2 was developed and applied to a variety of samples. The A2 precipitins were produced by a heifer infected with B. abortus strain 2308, cattle vaccinated with killed B. melitensis strain H38 or live B. abortus strain 19 and by a dog infected with B. canis. Precipitins were also detected in the second International Standard for anti-Brucella abortus serum, in several anti-B. canis sera and at low levels in one anti-B. ovis serum tested. Antisera produced in calves against Yersinia enterocolitica serotype 0:9 had no anti-A2 activity despite titers greater than or equal to 1/1024 and greater than or equal to 1/80 in standard Brucella agglutination and CF tests, respectively. The test for A2 precipitins lacked specificity as weak reactions were obtained with five of 295 sera from brucellosis-free herds. This test was relatively insensitive, detecting precipitins in only 16 of 24 sera from infected cattle and 27 of 54 sera positive by complement fixation and enzyme labelled antiglobulin tests performed with whole cell and smooth lipopolysaccharide antigens, respectively. The A2 precipitins were detected in nine sera from five cattle, in two infected herds, which were negative by agglutination and complement fixation tests.  相似文献   

4.
布鲁氏菌弱毒疫苗粘膜免疫及检测方法的研究   总被引：1，自引：0，他引：1  
为研究布鲁氏菌弱毒疫苗粘膜免疫及其检测方法,本实验采用粘膜点眼途径对健康母羊接种布鲁氏杆菌猪2号疫苗(S2)、牛19号疫苗(A19)和羊强毒株(M16),筛选布鲁氏杆菌病鉴别检测方法。将12月龄～14月龄母羊60只随机分为3组,以常规疫苗推荐剂量进行半量粘膜点眼接种。采集血液、淋巴、脏器进行布鲁氏菌病血清学检测和细菌学分离以及PCR检测。结果表明:布鲁氏菌弱毒疫苗抗体水平持续6个月,其中血清学的试管凝集试验、半胱氨酸凝集试验与补体结合试验的阳性符合率达到100%。细菌分离期为6个月,乳腺、乳腺淋巴、髂淋巴分离率较高;而强毒株M16的抗体水平和细菌分离持续12个月以上。结果显示以常规血清学和细菌学检测方法在点眼免疫布鲁氏菌S2、A19苗6个月后可以进行野毒感染和疫苗免疫畜的鉴别诊断。  相似文献   

5.
The level of non-specific reactions found in the brucellosis serology of ruminants in New Zealand was very low until July 1992. This changed when, in an export consignment of 1071 deer, 35% reacted in the Brucella abortus tube agglutination test with titres varying from 50 to 200 IU. The reactors were also positive in the Rose-Bengal agglutination test and most of them reacted in the complement fixation test with titres varying from 10 to 80 IU. Yersinia enterocolitica 0:9 was later isolated from one deer of this consignment. It was the first isolate of this serotype recovered in New Zealand from an animal. Shortly after, false reactors occurred more frequently than before in sera from Brucella abortus accredited free cattle herds. As the involvement of Yersinia enterocolitica 0:9 was suspected in these cases, faecal samples from reactors and in-contact animals were cultured for this organism. Yersinia enterocolitica 0:9 was isolated from nine of 19 herds showing one or more false Brucella abortus seroreactions. Prior to 1990, Yersinia enterocolitica serotype 0:9 had not been isolated in New Zealand, despite the recovery of a number of other bio- or serotypes of the organism from humans and animals. From 1990 onward, serotype 0:9 began to be isolated from human faecal samples with increasing frequency. Since the first isolations from deer and cattle in 1992, it has now also been recovered from a cat and an alpaca and from cattle without any association with false positive Brucella abortus reactions. All serotype 0:9 isolates were of biotype 2.  相似文献   

6.
将布氏杆菌分不同剂量皮下注射接种于健康牛,于接种后45 d剖杀,采集相关的器官组织制备组织切片,应用免疫组化方法检查细菌在牛体内的分布。结果显示,肝脏、脾脏、肾脏、淋巴结出现强阳性信号;在高剂量注射的牛体内,肺脏、颌下淋巴结出现阳性细胞;心脏和健康牛组织呈阴性反应。阳性信号主要位于感染细胞的胞质中,偶尔出现在细胞核内。  相似文献   

7.
Brucella spp. are Gram-negative, facultative, intracellular coccobacilli that are pathogenic to a variety of mammals, including ruminants and humans. The conventional serological test for diagnosing brucellosis in cattle in Korea is the standard tube agglutination test. However, agglutination tests sometimes give false-positive results due to cross-reactions with other pathogens. The outer membrane proteins of Brucella species have been extensively studied for their immunogenicity and serodiagnostic applications. However, an application of B. abortus OMPs for serodiagnosis has not been successfully established. In this study, cloning and expression of B. abortus Omp28, a group 3 antigen, were accomplished by PCR amplification cloning into a pMAL expression system, and purification of a recombinant Omp28 (rOmp28). The immunogenicity of rOmp28 was confirmed by Western blot analysis with Brucella-positive bovine serum. To determine whether rOmp2 has a potential benefit for use in the serodiagnosis of bovine brucellosis, rOmp28-based ELISA and latex bead agglutination test were performed. B. abortus positive (n=122) or negative (n=88) from TAT were positive (118/122, 96.7%) or negative (84/88, 95.4%) in ELISA and were positive (94/122, 77%) or negative (71/88, 81.7%) in that the latex bead agglutination test, respectively. The sensitivity, specificity and accuracy were 96.7, 95.4, 96.2% in ELISA and 77, 80.6, 78.5% in latex bead agglutination test, respectively. These findings suggest that the rOmp28 of B. abortus might be a good candidate for developing serological diagnostic tools for bovine brucellosis.  相似文献   

8.
The standard agglutination tests for detecting antibody to Brucella abortus were modified by addition of chelating agents (EDTA and EGTA) to the antigens. Approximately 80% of "singleton" agglutination test reactions, negative on the diagnostic complement fixation test, obtained with cattle sera were eliminated while no decrease in titer was apparent when sera from B. abortus infected or vaccinated cattle were tested.  相似文献   

9.
Abstract. This study reports the detection of antibodies against Brucella abortus and B. canis in wild neotropical carnivores kept in captivity in three zoos in northeastern Brazil. A total of 42 serum samples were examined, 17 from coatis (Nasua nasua), eight from crab-eating raccoons (Procyon cancrivorus), three from crab-eating foxes (Cerdocyon thous), three from hoary foxes (Lycalopex vetulus), two from little spotted cats (Leopardus tigrinus), five from tayras (Eira barbara), two from greater grisons (Galictis vittata), and two from neotropical river otters (Lontra longicaudis). The Rose-Bengal test and complement fixation test (CFT) were performed to detect anti-Brucella spp. antibodies, whereas the agar gel immunodiffusion test (AGID) was employed to detect anti-B. canis antibodies. The overall seroprevalence varied by species and by test; in addition, CFT and AGID seemed better able to detect antibodies against B. abortus and B. canis, respectively. This is the first study on the presence of anti-Brucella spp. antibodies in captive carnivores from Brazil, as well as the first report of antibodies to Brucella spp. in coatis, crab-eating raccoons, hoary foxes, little spotted cats, tayras, and greater grisons.  相似文献   

10.
为建立区分猪种布鲁菌S2疫苗株接种奶牛与布鲁菌自然感染奶牛,BLAST比对分析羊种、牛种、猪种、犬种、沙林鼠种和绵羊种6种布鲁菌基因序列,发现repA—related基因是猪种布鲁菌与牛种及羊种布鲁菌的差异基因。设计引物PCR扩增获得repA-related基因片段,克隆并原核表达得到了布鲁菌repA—related融合蛋白,以repArelated蛋白建立间接EI．IsA检测方法。用repA—related蛋白间接ELISA检测猪种s2疫苗株接种动物血清为阳性,检测牛种和羊种布鲁菌自然感染动物血清为阴性。repA—related蛋白间接EusA能从试管凝聚实验（SAT）及常规ELIsA检测阳性的奶牛血清样本中,区分出s2疫苗接种牛与牛种布鲁菌感染牛。  相似文献   

11.
布鲁菌属革兰氏阴性兼性胞内寄生菌,能感染多种宿主动物和人。该属可分为6个典型种,包括羊种、牛种、猪种、沙林鼠种、绵羊附睾种以及犬种布鲁菌等。此分类是基于其致病性以及宿主偏好性的差异划分。尽管6个种通过传统表型试验能区分,但布鲁菌种内采用DNA-DNA杂交证明DNA同源性高度一致（相似性大于90%）。因此有人提议布鲁菌由单一种组成,即布鲁菌属中只有羊种布鲁菌,其他种都是羊种菌的生物亚型之一。然而基于其他分子技术的基因分型表明其DNA多态性表现明显,说明目前对这个种的分型还是比较准确。而最近分离的海洋种布鲁氏菌分离株（鳍型和鲸型）采用传统分型标准和一些特异的分子标记也证明这种分型比较正确。本文对目前布鲁菌种属进化和分类学进行综述,希望对研究其进化和分类有所帮助。  相似文献   

12.
A preliminary serological study of 366 household dogs in Lagos and Ibadan, southwestern Nigeria, was carried out to determine antibodies due to exposure to Brucella abortus and B. canis, using the rose bengal test (RBT) and the rapid slide agglutination (RSA) test, respectively. Results showed that 5.46 % (20/366) and 0.27 % (1/366) of the dogs screened were seropositive to B. abortus and B. canis, respectively. Of all dogs, 36 had a history of being fed foetuses from cows and 11 (30.6 %) of these tested positive in the RBT. Our findings, although based on a limited sample size and a dearth of clinical details, revealed that dogs in Nigeria may be infected with Brucella spp. given the wide range of risk factors. Further studies are recommended to elucidate the epidemiology of brucellosis in dogs and its possible zoonotic consequences in the country.  相似文献   

13.
Horses at a veterinary teaching hospital and a slaughterhouse were surveyed for antibodies to Brucella abortus, B canis and Actinobacillus equuli. Four of the 141 hospitalised horses and none of the 73 slaughtered horses had titres of 1:100 or greater to B abortus. Six horses of both populations reacted to the card test. One was culture positive. A card test using B canis antigen was positive in 38 per cent of the sera from hospitalised horses and all of the slaughtered horses. Twenty (27.4 per cent) of the latter group had high tires in a tube agglutination test. High titres could not be reduced by 2-mercaptoethanol serum treatment. The titres appeared to be associated with advanced age but not to sex. Adsorption of sera with B canis did not affect titres to A equuli but the reverse was true.  相似文献   

14.
Seven of 18 elk on a deer farm were found by the official Rose‐Bengal agglutination test (RBT) and tube agglutination test to be brucellosis reactors/suspects. Evaluation with the competitive ELISA (C‐ELISA) and the fluorescence polarization assay (FPA) tests revealed that six and five sera were positive respectively. The seven reactors/ suspects were slaughtered and their blood and tissues were collected. Brucella species could be isolated from three of the slaughtered animals, with nine isolates being obtained from the popliteal, supramammary and submandibular lymph nodes, vaginal discharge, mammary tissue and spleen. Brucella genus‐specific PCR based on 16S rRNA and AMOS‐PCR, which is specific for differential Brucella species, revealed that all nine isolates were Brucella abortus. These nine were further confirmed to be B. abortus biovar 1 by classical biotyping scheme assays. This is the first report of an outbreak of brucellosis in domestic elk in Korea. Our observations suggest that deer should be included in the routine Brucella surveillance programme for the effective control and prevention of brucellosis in Korea.  相似文献   

15.
Cell-mediated immune responses in cattle adult-vaccinated with Brucella abortus strain 19, cattle infected with B abortus field strain, and nonexposed cattle were studied by an in vitro lumphocyte-stimulation test (LST). Lymphocytes were prepared from peripheral bovine blood by the Ficoll-diatrizoate technique, and results were assayed for [3H]thymidine incorporation into DNA by liquid scintillation spectrometry. Serotests and bacteriologic isolation attempts were conducted simultaneously with LST. Lymphocytes from cattle infected with field strains had significantly (P = 0.01) higher specific lymphocyte-stimulation inexposed controls. The LST, the serum standard-tube agglutination test (STT), the Rivanol (RIV) test, and the complement-fixation (CF) test correctly classified cattle from which field strains and strain 19 of B abortus were isolated. The LST was negative in cattle vaccinated with B abortus strain 19 (nonshedding), but the three serotests had many false-positive reactions. The CF test had the least false-positive reaction, followed by the RIV test, and the STT was the least specific. Well before the three serotests became positive, the LST was positive in samples from some cattle during the incubation period of the infection. There was little or no correlation between cell-mediated immune responses (as measured by LST) and serum antibody responses (as measured by STT, RIV test, and CF test) in vaccinated but culture-negative cattle and in some nonvaccinated cattle during the incubation period.  相似文献   

16.
During the first half of the twentieth century, widespread regulatory efforts to control cattle brucellosis due to Brucella abortus in the Union of Soviet Socialist Republics were essentially non-existent, and control was limited to selective test and slaughter of serologic agglutination reactors. By the 1950s, 2-3 million cattle were being vaccinated annually with the strain 19 vaccine, but because this vaccine induced strong, long-term titers on agglutination tests that interfered with identification of cattle infected with field strains of B. abortus, its use in cattle was discontinued in 1970. Soviet scientists then began a comprehensive program of research to identify vaccines with high immunogenicity, weak responses on agglutination tests and low pathogenicity in humans, as a foundation for widespread control of cattle brucellosis. While several new vaccines that induced weak or no responses on serologic agglutination tests were identified by experiments in guinea pigs and cattle, a large body of experimental and field studies suggested that the smooth-rough strain SR82 vaccine combined the desired weak agglutination test responses with comparatively higher efficacy against brucellosis. In 1974, prior to widespread use of strain SR82 vaccine, over 5300 cattle farms across the Russian Federation were known to be infected with B. abortus. By January 2008, only 68 cattle farms in 18 regions were known to be infected with B. abortus, and strain SR82 continues to be the most widely and successfully used vaccine in many regions of the Russian Federation.  相似文献   

17.
Brucellosis is a highly infectious disease which is diagnosed using serological and microbiological methods. The objective of this study was to assess the viability of using conventional and real-time PCR assays as potential diagnostic tools for the detection of Brucella abortus in naturally infected cows. PCR assays that amplify various regions of the Brucella genome, IS711 genetic element, 31kDa outer membrane protein and 16S rRNA, were optimised using nine known Brucella strains. Real-time PCR was used to examine the detection efficiency of the IS711 assay which was estimated at 10 gene copies. Milk, blood and lymph tissue samples were collected from naturally infected animals. B. abortus was not detected in blood samples collected from naturally infected cows by conventional or real-time PCR, but was detected in a proportion of the culture-positive milk (44%) and lymph tissue (66% - retropharyngeal, 75% - supramammary) samples by the same methods. There was no difference between PCR and bacteriological detection methods. It is unlikely that conventional or real-time PCR will supersede current diagnostic methods for detection of B. abortus in clinical samples.  相似文献   

18.
Genomic DNA from reference strains and biovars of the genus Brucella was analyzed using pulsed-field gel electrophoresis (PFGE). Fingerprints were compared to estimate genetic relatedness among the strains and to obtain information on evolutionary relationships. Electrophoresis of DNA digested with the restriction endonuclease XbaI produced fragment profiles for the reference type strains that distinguished these strains to the level of species. Included in this study were strains isolated from marine mammals. The PFGE profiles from these strains were compared with those obtained from the reference strains and biovars. Isolates from dolphins had similar profiles that were distinct from profiles of Brucella isolates from seals and porpoises. Distance matrix analyses were used to produce a dendrogram. Biovars of B. abortus were clustered together in the dendrogram; similar clusters were shown for biovars of B. melitensis and for biovars of B. suis. Brucella ovis, B. canis, and B. neotomae differed from each other and from B. abortus, B. melitensis, and B. suis. The relationship between B. abortus strain RB51 and other Brucella biovars was compared because this strain has replaced B. abortus strain 19 for use as a live vaccine in cattle and possibly in bison and elk. These results support the current taxonomy of Brucella species and the designation of an additional genomic group(s) of Brucella. The PFGE analysis in conjunction with distance matrix analysis was a useful tool for calculating genetic relatedness among the Brucella species.  相似文献   

19.
Characterization of an atypical biotype of Brucella abortus.   总被引：2，自引：1，他引：1       下载免费PDF全文
Brucella abortus strains were isolated from bovine tissue and milk samples from seven Ontario herds. The isolates were characterized by colonial morphology, requirement of CO2 for growth, lysis by Tbilisi phage, biochemical tests and agglutination in monospecific sera. They resembled B. abortus biotype 2 (on the basis of sensitivity to thionin and basic fuchsin) and biotype 4 (on the basis of agglutination with anti-Brucella "M" but not anti-Brucella "A" absorbed sera). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of these isolates and B. abortus biotypes 1, 2 and 4 showed similar profiles. Immunoblots with anti-A and anti-M absorbed sera showed different antigenic regions reacting with the specific sera and also confirmed that the atypical B. abortus isolates were serologically similar to biotype 4.  相似文献   

20.
Although some ELISA tests using cytoplasmic or outer membrane antigens of Brucella have been developed to improve the diagnosis of canine brucellosis, the performance of these assays has not been compared. In the present study three ELISA tests using lipopolysaccharide (LPS)-free cytoplasmic proteins (CPs) of Brucella abortus, the lumazine synthase (LS) of Brucella spp. or a hot-saline (HS) extract of Brucella canis containing outer membrane antigens were used to test sera from dogs with suspected or confirmed brucellosis (n=36) and from dogs with pathological conditions other than brucellosis (n=212). In the first group the proportion of positive results was 92, 92 and 81% for the ELISAs with HS, CP and LS, respectively, and 94% of the samples were positive by at least one ELISA test. Three dogs that were negative by agglutination (2ME-RSAT) had a positive result by at least one ELISA, and this discrepancy was attributed to the lower analytical sensitivity of agglutination tests. This hypothesis was confirmed by a serological follow-up of seven dogs recently infected with B. canis in three of which the illness was diagnosed earlier by one or more ELISA tests than by 2ME-RSAT. Among dogs having pathological conditions other than brucellosis, specificities were 94.3, 96.7 and 96.7% for the ELISAs with HS, CP and LS, respectively. This study shows that HS-ELISA and CP-ELISA are highly specific and sensitive for the diagnosis of canine brucellosis and can detect the infection by B. canis shortly after the exposure to the pathogen.  相似文献   

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