• 한국동물위생학회지
• /
• 제34권2호
• /
• pp.153-157
• /
• 2011

In order to investigate serum antibodies for detection of brucellosis in pigs, a total of 1208 sera were tested by Rose Bengal test (RBT), the standard tube agglutination test (STAT) and competitive ELISA (cELISA). The sera were collected from pigs of Gyeonggi, Chungnam, Chungbuk, Jeonnam and Jeonbuk, provinces during the period 2002 to 2004. All the sera were screened by RBT, and were confirmed by STAT and cELISA. Among 1208 sera, 26 sera (2.2%) were positive in screening test. All the 26 positive sera were positive by STAT, while all the sera were negative by cELISA. On the basis of this study, farmed pigs may be exposed to Brucella species. Furthermore, these results suggest that establishment of diagnoses for detection of porcine brucellosis is necessary.

• 대한수의학회지
• /
• 제38권2호
• /
• pp.345-352
• /
• 1998

The diagnosis of brucellosis is currently based on serological and microbiological tests. However, the microbiological isolation and identification have several disadvantages such as time-consuming and laborious, and the serological methods have been reported to cross-react with antigens other than those from Brucella spp. To develop a sensitive and rapid diagnostic method for detection of Brucella species, the genus-specific primers were designed and synthesized from the sequence of gene encoding a 31kDa cell surface protein(BCSP) and a 36kDa outer membrane protein(OMPB) of B abortus. The amplified 711bp and 982bp DNA fragments were only visible in each species of Brucella by PCR method using the BCSP and OMPB primers, respectively. However, PCR product was not obtained with DNA from other Gram-negative bacteria. As little as 1pg of the B abortus genomic DNA could be detected by this PCR method. Using the PCR technique, semen samples from 185 bulls of Brucella-seronegative herds in Cheju island were examined for comparison of this PCR method with conventional methods in 1995. The semen samples from 5 bulls were positive by culture method and PCR, and one was positive and 5 were suspect by semen plasma agglutination test. However, the semen samples obtained from 177 bulls were negative by semen plasma agglutination, culture and PCR methods in 1996. The results of comparison tests suggested that PCR was a better test than agglutination test against semen of bulls. This study indicated that the PCR technique was a valuable for the diagnosis of bovine brucellosis, particulary in bull semens.

• Journal of Microbiology and Biotechnology
• /
• 제30권4호
• /
• pp.497-504
• /
• 2020

For control of brucellosis in small ruminants, attenuated B. melitensis Rev1 is used but it can be virulent for animals and human. Based on these aspects, it is essential to identify potential immunogens to avoid these problems in prevention of brucellosis. The majority of OMPs in the Omp25/31 family have been studied because these proteins are relevant in maintaining the integrity of the outer membrane but their implication in the virulence of the different species of this genus is not clearly described. Therefore, in this work we studied the role of Omp31 on virulence by determining the residual virulence and detecting lesions in spleen and testis of mice inoculated with the B. melitensis LVM31 mutant strain. In addition, we evaluated the conferred protection in mice immunized with the mutant strain against the challenge with the B. melitensis Bm133 virulent strain. Our results showed that the mutation of omp31 caused a decrease in splenic colonization without generating apparent lesions or histopathological changes apparent in both organs in comparison with the control strains and that the mutant strain conferred similar protection as the B. melitensis Rev1 vaccine strain against the challenge with B. melitensis Bm133 virulent strain. These results allow us to conclude that Omp31 plays an important role on the virulence of B. melitensis in the murine model, and due to the attenuation shown by the strain, it could be considered a vaccine candidate for the prevention of goat brucellosis.

• 한국동물위생학회지
• /
• 제32권4호
• /
• pp.335-341
• /
• 2009

Canine brucellosis produce abortions and infertility in dogs and is currently diagnosed by serological methods such as rapid slide agglutination test with 2-mercaptoethanol (2-ME RSAT) and immunochromatographic assay (ICA). Bacterial isolation is considered gold standard for Brucella diagnosis and the polymerase chain reaction (PCR) is an alternative method to bacterial isolation. A total of 36 whole blood samples were collected from dogs reared in area of Chuncheon and were subjected to serology (2-ME RSAT and ICA for B. canis, Rose Bengal test and C-ELISA for B. abortus), blood culture and 3 types of PCRs (BSCP31, 16s rRNA, and OMP-2). All blood samples were negative by serology and blood cultures. The BCSP31 and the OMP-2 PCR detected 5 samples were positive whereas the 16S rRNA PCR detected all samples were negative as serological methods and blood culture did. From the results observed in the present study, we conclude that 16S rRNA PCR could be used for direct PCR for canine blood samples.

• Journal of Preventive Medicine and Public Health
• /
• 제38권4호
• /
• pp.482-488
• /
• 2005

Objectives : We conducted an investigation on 14 cases of brucellosis in Gyeongsangbuk-do during 2003-2004 to understand the source of infection and the transmission routes of brucellosis. Methods : The authors visited the each of the health centers and we examined the patients, their written epidemiologic questionnaire and the occurrence of bovine brucellosis. We visited the patients' living and work areas, and we examined their occupations, the date they developed symptoms, the progress of their symptoms, whether or not they were treated, their current status, whether or not they consumed raw milk and raw meat, and if their work was related to cattle breeding and the related details. We reviewed the results of the blood tests and medical records and we examined the cattle's barn. Results : There were 3 patients in 2003 and 11 patients in 2004. All of their brucella antibody titer exceeded 1:160. The patients' symptoms were fever, myalgia, malaise, chills and an influenza-like illness, but the clinical signs were absent on the medical records. Brucella abortus were cultured from 3 of the patients' blood samples. Conclusions : When the authors discovered the transmission routes, they were divided into 4 different sorts. The first route was related to cattle birth such that patients touched the calves or placentas that were infected with the Brucella species. The second route was related to performing artificial insemination on the cattle and the semen that was used for artificial insemination. The third route was due to the ingestion of raw meat and milk. The last route was due to sexual intercourse between the patients.

• Journal of Microbiology and Biotechnology
• /
• 제20권12호
• /
• pp.1750-1755
• /
• 2010

The MLVA assay is known to have a high ability to identify and discriminate Brucella species, so that it can be used as an epidemiological tool to discriminate Brucella isolates originating from restricted geographic sources. In this study, the genetic profiles of 38 B. abortus isolates from humans were analyzed and compared with genotypes from animal isolates in South Korea. As a result, it was found that they did not show high genetic diversity and were compacted. They were clustered together with animal isolates, showing a significant correlation to regional distributions. With its ability to prove a significant genetic correlation among B. abortus isolates from animals and humans in South Korea, the MLVA assay could be utilized as part of a program to control and eradicate brucellosis, one of the major zoonoses. This study represents the first data of genetic correlation of B. abortus isolates from humans and animals in South Korea.

• Journal of Microbiology and Biotechnology
• /
• 제26권3호
• /
• pp.603-609
• /
• 2016

Brucellosis affects a wide range of host species, including humans and many livestock animals. Chronic infections of the disease make antibiotic treatment costly, and the current vaccine used in livestock has not been approved for human use. This study investigated the possible use of the Brucella abortus outer membrane protein A (OmpA) as a candidate subunit vaccine in an infected mouse model. The ompA gene was cloned and overexpressed, and the recombinant OmpA (rOmpA) protein fused to maltose binding protein (MBP) was purified in Escherichia coli. Immunogenicity was verified through western blotting, and mice were immunized and challenged to evaluate its protective effect. Mice treated with rOmpA exhibited induced humoral and host cell-mediated responses, with a significant increase in immunoglobulin G (IgG1 and IgG2a) and cytokine levels, especially TNF-α and IL-12, compared with the control groups treated with either MBP or PBS. In conclusion, rOmpA should be highly considered as a future subunit vaccine for brucellosis, and further studies regarding rOmpA and its protective ability are suggested.

• 한국군사과학기술학회지
• /
• 제22권4호
• /
• pp.575-580
• /
• 2019

In biological warfare, it is important to identify biological agents for proper treatment. We focused on developing a real-time RT-PCR kit that can detect multiple species of biological agents. AccuPower(R) Biothreat Real-Time RT-PCR Kit(v3.0) could detect Bacillus anthracis, Yersinia pestis, Vibrio cholerae, Francisella tularensis, Salmonella typhi, Rickettsia prowazekii, Variola virus, Hantaan virus, Yellow fever virus, Brucella spp., Shigella dysenteriae in a single reaction. The results showed that the kit was verified to be able to detect at least 0.005 ng of nucleotide and 10,000 CFU/ml of bacteria. Therefore, the kit is expected to be used as a rapid and sensitive detection kit for 11 species of biological agents within 2 hours.

• 한국동물위생학회지
• /
• 제31권4호
• /
• pp.493-496
• /
• 2008

Brucella spp. are small, non-motile Gram-negative coccobacilli known to cause disease in a number of vertebrate species including humans and brucellosis is one of the world's major zoonoses, alongside bovine tuberculosis and rabies. There are about 33.55 million goats and 1.16 million sheep in Bangladesh. The sheep and goats can significantly play an important role in the economic well being of the resource-poor farmer in Bangladesh. Sexually matured 362 female small ruminants(300 goats and 62 sheep) were examined. Approximately 3-5 ml of blood was collected from the jugular vein of each animal and sera samples were prepared. Samples were then tested for brucellosis by using Rose Bengal test(RBT), plate agglutination test(PAT) and tube agglutination test(TAT). Among 362 small ruminants, irrespective of species(sheep or goat), diagnosed highest in TAT, 2.21%(n=8) and lowest both by RBT & PAT, 1.93%(n=7) and it is concluded that TAT is superior than RBT and PAT.

• 대한임상검사과학회지
• /
• 제38권1호
• /
• pp.45-53
• /
• 2006

Anaerobic black-pigmented bacteria have been implicated in the endodontic infections. This group of microorganisms includes Porphyromonas endodontalis, P. gingivalis, Prevotella intermedia, and Prevotella nigrescens. The organisms display a wide variety of virulence factors that may be pertinent to acute endodontic infections. The aim of this study was to identify P. endodontalis, P. gingivalis, P. intermedia, and P. nigrescens by using the special potency disk test, filter paper spot test, 16S rRNA gene-directed PCR, and API 32A system. Microbial samples were collected from root canals of 33 intact teeth with necrotic pulp and apical periodontitis. Conventional laboratory methods were used to identify the strains of anaerobic black pigmented bacteria. Eighteen out of 33 samples were positive for the growth of black-pigmented bacrteria. Five colonies were cultured from each pure cultured colony from Brucella agar plates. Seventy seven colonies were positive for the growth of black-pigmented bacteria. Thirty three out of 77(42.8%) were identifed as P. nigrescens, 10 out of 77(13%)were P. gingivalis, 6 out of 77(7.8%) were P. endodontalis, 10 out of 77(13%) were P. intermedia. On the contrary the reference strains of P. nigrescens, experimental strains of P. nigrescens were susceptible to kanamycin in the special potency disk test. We concluded that after rapid presumptive identification methods, such as the special potency disk test and filter paper spot test were done, 16S rRNA gene PCR and API 32A test would be accurate detection methods for black-pigemented bacteria.