Brucella canis osteomyelitis in two dogs with total hip replacements

Brucella canis was isolated from the cement or bone surrounding a hip prosthesis after total hip replacement was performed for treatment of hip dysplasia in 2 dogs. Lameness or signs of infection were not evident for 9 and 16 months after surgery. Osteomyelitis surrounding the prostheses was detected radiographically only after the lameness developed. The origin of the B canis infection in the 2 dogs was believed to be hematogenous because of the biologic behavior of this organism and because of the duration of excellent limb function after hip replacement. A slide agglutination test for B canis should be performed as a screening test on any canine total hip candidate when the anamnesis and physical examination indicate that the dog may have been exposed to or infected with B canis.     

The first detection of brucella canis in cattle in the Republic of Korea

Twenty mammary lymph node samples were collected from cattle on a farm in the Republic of Korea. These cattle were serologically negative for Brucella by tube agglutination test (≤ 1:50) and serum agglutination test (≤ 1:50). Out of 20 lymph node samples, two samples were positive for Brucella growth on Brucella agar as well as blood agar. Tests for urease, hydrogen sulphide and reactions against monospecific sera A and M indicated that these two isolates (No. 15 and 16) belong to the genus Brucella. Genus specific, AMOS (abortus, melitensis, ovis, suis) and Bruce-ladder multiplex polymerase chain reaction (PCR) assays confirmed the Brucella isolates as either a B. abortus or a B. canis strain. This is the first report of the occurrence of a B. canis infection in cattle in Korea. More survey data are needed to determine whether B. canis is a significant aetiology in the cases of cattle brucellosis in Korea.     

A serologic survey of a population of Georgia dogs for Brucella canis and an evaluation of the slide agglutination test.

In a serologic survey of stray and pet dog populations of Georgia, serums were screened for Brucella canis antibodies, using the slide agglutination test. If results were positive, B canis antibody titers were determined, using the standard tube agglutination test. The stray dogs had significantly (P less than 0.01) higher titers than did the pet dogs. The reactor rate was 58% higher for the slide agglutination test than for the tube agglutination test. The manufacturer's evaluation of the slide agglutination test was based on a comparison of the serologic results of that test with those of the tube agglutination test, using a comparative method that permitted the results to be interpreted as 99% agreement between the 2 tests. Reevaluation of the manufacturer's data by a different method indicated that the slide agglutination test is very accurate when the results are negative (99.7% specific) but less so when the results are positive (62.5% sensitive).     

A rapid slide agglutination test for the serodiagnosis of Brucella canis infection that employs a variant (M-) organism as antigen

An improved antigen is described for use in the serodiagnosis of canine brucellosis. The novel antigen employs a less mucoid (M-) variant of B. canis. The replacement of Brucella ovis with B. canis (M-) cells as antigen in slide agglutination tests would increase accuracy by reducing the rate of false positive results from ca. 50% to ca. 10%. Rose bengal stained B. canis (M-) cells are slightly more sensitive than the commercially available Brucella ovis as slide test antigens. Both test procedures require brief (less than 1 min) reaction with 2-mercaptoethanol in order to reduce the rate of false positive reactions.     

BRUCELLA CULTURES TYPED BY THE WHO BRUCELLOSIS CENTRE AT THE COMMONWEALTH SERUM LABORATORIES, MELBOURNE

Results of the typing of Brucella cultures received at the WHO Brucellosis Centre, CSL, Melbourne, from 1968-1976 are presented. The distribution of the biotypes of cultures recovered throughout Australia is shown on a host and State basis and atypical cultures are discussed. Cultures identified from Australia were Br. abortus, biotypes 1, 2 and 4 and Strain 19; Br. suis, biotype 1 and Br. ovis. Br. melitensis biotypes 1, 2 and 3 were recorded only as exotic human infections from the Mediterranean area and from laboratory infections. Br. abortus, biotype 1 was the most common bovine and human isolate and was found in horses, a goat and a sheep. There was a low incidence of Br. abortus, biotype 2 in cattle and it was found in 1 horse. Br. abortus, biotype 3 was not found in Australia but was submitted from Tanzania. Br. abortus, biotype 4 was rare in cattle and was found once in a horse. Br. abortus, Strain 19 was found occasionally in cattle and once from a test guinea pig. Br. suis, biotype 1 was found in both man and pigs in Queensland and New South Wales whereas biotype 3 was isolated only in New Guinea from pigs and cattle. Br. ovis was submitted from 3 States. Br. canis has not been found in Australia.     

The identity, distribution and epizootiological significance of brucella isolates in Australia, 1981 to 1985

Results listing the identification of brucella isolates received by the National Brucellosis Reference Centre, National Biological Standards Laboratory, Canberra from 1981 to 1985 are presented. The distribution of brucella species and biotypes is shown on a host and state basis. Cultures isolated in Australia were identified as Brucella abortus biotypes 1, 2 and 4, and Strain 19; B. suis biotype 1, and B. ovis. B. melitensis biotype 3 was recovered from man infected in the Mediterranean area. B. abortus biotype 1 was the most frequent isolate. Atypical cultures isolated from cattle included B. suis biotype 1, and erythritol-utilising mutants of Strain 19. The epizootiological implications of these findings are discussed in relation to their impact on the national campaign to eradicate bovine brucellosis.     

一株猪源链球菌的分离鉴定及生物学特性研究

从送检的一份疑似链球菌病病死猪病料中分离出1株细菌,通过对分离菌株进行细菌的培养特性、生化试验、玻片凝集试验及PCR分型鉴定,确定为2型猪链球菌,动物致病性试验表明,1×10⁸ CFU/mL的剂量可致死小白鼠,对该分离菌株的毒力因子进行PCR检测发现,溶菌酶释放蛋白(mrp)、胞外因子(ef)和溶血素(sly)均为阳性。此菌的分离鉴定为自家灭活菌苗的制备提供了候选菌株。     

Rapid detection of Streptococcus suis serotype 2 in weaned pigs

A survey to detect Streptococcus suis serotype 2 in 1,716 weaned pigs was done in Quebec. Forty-nine sow herds were included in this survey: in 26 herds, S suis serotype 2 had been isolated during the preceding 12 months and in 23 herds (control), the organism had not been detected during a previous study. Swab specimens of the nasal cavity and tonsils of pigs were obtained for bacteriologic culture, and S suis serotype 2 was easily detected by the use of brain-heart infusion agar containing a Streptococcus-selective supplement and 5% goat antiserum raised against S suis serotype 2. After measurement of the diameter of the precipitation zone of 539 isolates, a slide agglutination test was performed to identify the S suis serotype 2 isolates. The mean precipitation zone diameter obtained for group S suis serotype 2 was larger (P less than 0.001) than that for the group designated as "others". With slide agglutination test results as reference and on the basis of discriminant analysis to stimulate detection of S suis serotype 2, 93.1% of all isolates were correctly classified, using the precipitation zone diameter as unique classification criterion. Relative specificity was 94.5% and relative sensitivity was 88.7%. Use of the precipitation zone diameter on a quantitative basis led to the proposal of a simple and reliable technique to screen swine herds for S suis serotype 2 in weaned pigs. Nasal and tonsillar swab specimens were obtained and analyzed concurrently for S suis serotype 2. The organism was found in both sites in only 20.4% of 103 carrier pigs.(ABSTRACT TRUNCATED AT 250 WORDS)     

布鲁菌S2疫苗株免疫牛与牛种、羊种布鲁菌自然感染牛ELISA鉴别诊断方法的建立

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。     

Seroprevalence of Brucella abortus and B. canis in household dogs in southwestern Nigeria: a preliminary report

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.     

Immunologic properties of soluble materials from Brucella species.

The purpose in this study was to examine the immunogenic properties of various preparations of aqueous ether extracts of Brucella suis and Brucella canis. The B suis strain 3b and B canis strain RM-6-66 were grown on tryptose agar, and aqueous ether extracts were prepared from the cells. The ether was removed, and the extracts were clarified by centrifugation for 10 hours at 144,000 X g and fractionated by gel chromatography. The B suis endotoxin-containing precipitate, obtained from aqueous ether extracted material by ethanol precipitation, and fraction 1, prepared from ultracentrifugal supernate by column chromatography, protected mice against homologous infection. The B canis aqueous ether-extracted material also protected mice against B suis infections.     

2型猪链球菌的血清学鉴定

本试验首次证实了２型猪链球菌在我国的存在，采用玻片凝集、琼脂扩散及毛细管沉淀等３项试验，对１９９０年以来广东某猪场断奶仔猪暴发流行的败血症、脑膜炎及关节炎病猪中分离的１５株链球菌进行了血清学鉴定。结果表明：１５株链球菌均为２型猪链球菌；用高压法提取抗原进行沉淀反应，其结果比酸热法提取抗原具有简便、敏感、特异性强的特点。     

New Bruce-ladder multiplex PCR assay for the biovar typing of Brucella suis and the discrimination of Brucella suis and Brucella canis

Rapid and specific identification of Brucella suis at the biovar level is necessary because some of the biovars that infect animals are pathogenic for humans. None of the molecular typing methods described so far are able to discriminate B. suis biovars in a single test and differentiation of B. suis from Brucella canis by molecular approaches can be difficult. This article describes a new multiplex PCR assay, Suis-ladder, for fast and accurate identification of B. suis at the biovar level and the differentiation of B. suis, B. canis and Brucella microti. An advancement of the original Bruce-ladder PCR protocol which allows the correct discrimination of all known Brucella species is also described.     

Characterization of an atypical biotype of Brucella abortus.

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.     

Evaluation of cattle for experimental infection with and transmission of Brucella suis biovar 4

OBJECTIVE: To determine whether cattle can become persistently infected with Brucella suis biovar 4, whether the organism can be transmitted vertically or horizontally, and whether tests for bovine brucellosis are diagnostic. DESIGN: Observational study. ANIMALS: 24 pregnant cows and their calves and 6 bulls. PROCEDURE: Cows and bulls were housed separately in groups of 6 with each group consisting of 3 cattle experimentally infected with B suis biovar 4 and 3 na?ve animals. Cattle were observed for clinical signs daily; blood samples were collected weekly. Clotted blood from each sample was submitted for bacterial culture. Serum was tested with an indirect ELISA and the standard tube agglutination test (STAT), buffered plate agglutination test, brucellosis card test (BCT), and complemen't fixation test (CFT). Tissues collected at necropsy were submitted for bacterial culture and histologic examination. RESULTS: All 15 inoculated cattle seroconverted on 2 or more serologic tests, and bacteria were isolated from 4 inoculated cows at necropsy. There was no bacteriologic evidence of vertical or horizontal transmission, and none of the cattle developed clinical abnormalities or gross or histologic lesions. Results of the indirect ELISA were positive for all inoculated cattle. The other tests gave variable results; the CFT, STAT, and BCT yielded negative results for at least 1 of the 4 cattle from which the organism was isolated. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cattle-to-cattle transmission of B suis biovar 4 is unlikely. Serologic tests for bovine brucellosis should be used cautiously when attempting to identify cattle with rangiferine brucellosis, as they do not discriminate between the 2 diseases and vary in their ability to detect exposed cattle.     

Agglutinins to Brucella canis in stray dogs from certain counties in Illinois and Wisconsin.

In a serologic and blood culture survey for Brucella canis in 2,572 pound dogs from eight Illinois and Wisconsin counties, 173 (6.7%) were reactive in a presumptive slide test, 41 (1.5%) were reactive in a tube agglutination test, and 6 (0.2%) were positive in blood culture.     

Study of agglutinins to Brucella abortus, B canis and Actinobacillus equuli in horses

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.     

The bovine immune response to Brucella abortus IV. Studies with a double immunodiffusion test for antibody against A2.

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.     

Swine brucellosis in Indonesia

Brucella suis biotype 1 was isolated from 13.1% of the pigs slaughtered in Kapuk Jakarta, West Java and from 15.09% of the pigs slaughtered in Surabaya, East Java. The prevalence of B. suis by means of the Rose Bengal Plate Test, was 22.3% for West Java and 14.9% for East Java. The Rose Bengal Plate Test detected more B. suis infected animals (73% of the infected animals) than did the Complement Fixation Test (41%) and the Serum Agglutination Test (54.5%). The high infection rate is a potential health danger for the abattoir workers.     

A rapid agglutination assay for canine brucellosis using antigen coated beads

Brucella canis is the causative agent of canine brucellosis and facultative intracellular pathogen. The diagnosis of canine brucellosis is based on bacteriological examination and serological methods including agglutination and gel diffusion tests. In this study, crude antigens were extracted from B. canis using hot saline, coated on to latex beads and their usefulness in the serological diagnosis of canine brucellosis was examined. Mixing the antigen coated latex beads with the sera of dogs infected with B. canis produced clear agglutination, but this was not so for B. canis free dog sera. N-terminal amino acid sequence analysis of the crude hot saline extracts, showed that they contained copper-zinc superoxide dismutase, ribose ABC transporter and hypothetical protein of Brucella as antigens. A serological survey of canine serum samples conducted by means of an agglutination test using the antigen coated latex beads, showed that this method was more specific than the tube agglutination test using whole bacterial cell antigens. Although these results suggest that our method in which crude hot saline extracted antigens are coated on to latex beads would be useful in the serological diagnosis of canine brucellosis, we need further investigation using more serum samples to confirm the usefulness of our method.