• Title/Summary/Keyword: Brucella abortus

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Development of PCR assay for the detection of Brucella spp in bovine semen (종모우 정액중 Brucella균 신속 검출을 위한 PCR기법 개발)

  • Jung, Suck-chan;Jung, Byeong-yeal;Woo, Seong-ryong;Cho, Dong-hee;Kim, Jong-yeom;Kim, Woo-taek;Lee, Jung-mi;Park, Yong-ho;Baek, Byeong-kirl
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.345-352
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    • 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.

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Immune Modulation of Recombinant OmpA against Brucella abortus 544 Infection in Mice

  • Simborio, Hannah Leah Tadeja;Reyes, Alisha Wehdnesday Bernardo;Hop, Huynh Tan;Arayan, Lauren Togonon;Min, Wongi;Lee, Hu Jang;Lee, Jin Ju;Chang, Hong Hee;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.603-609
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    • 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.

The Safety of Brucella abortus strain RB51 in pregnant cattle (부루세라 RB51 백신의 임신소에 대한 안전성)

  • Kim, Jong-man;Woo, Sung-ryong;Lee, Ji-youn;Jung, Suk-chan;Jean, Young-hwa;Kim, Jae-hoon;Kweon, Chang-hee;Yoon, Young-dhuk;Yoo, Han-sang
    • Korean Journal of Veterinary Research
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    • v.43 no.4
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    • pp.667-675
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    • 2003
  • The safety of Brucella abortus strain RB51(SRB51) was investigated in dairy cows and Korean native cattle of 4~7th month of gestation. From experimentally inoculated cattle, 18 of 25 (72.0%) dairy cows, and 3 of 10 (30.0%) Korean native cattle were aborted or delivered premature fetus. There were no significant differences in the incidence of abortion depending on the inoculation route (intramuscular and subcutaneous) and dosages ($1{\times}10^9$, $2.8{\times}10^9$, and $4.0{\times}10^9$ CFU). The antibodies to the SRB51 were measured by a dot blot enzyme-linked immunosorbent assay. The highest titers to SRB51 were detected between 5~7 weeks after inoculation and the specitic antibody could be detected up to 28 weeks after inoculation. The SRB51 was isolated from amnio-allantoic fluid, bronchial lymph node, mammary gland, and supramammary lymph node in 5 of 25 dairy cows during 4 weeks after either abortion or delivery. Although SRB51 was isolated from 4 of 24 aborted fetus or normally delivered calves at parturition time, it was not isolated during 4 weeks afterward. Eleven of twentyfive dairy cows showed the endometritis and/or necrosis until 6 weeks after delivery, no lesions were seen at 8 weeks after delivery and uterus from control dairy cows. The results of present study revealed that SRB51 might induce the clinical signs of brucellosis in the pregnant cattle at 4~7th month of gestation.

Isolation of Brucella spp from sere-positive native bulls and calves below twelve months old (브루셀라병 혈청검사 양성 수소와 12개월령 이하 소에서의 균 분리 및 동정)

  • Ryu, Jae-Yun;Bun, Jung-Hyun;Lee, Hee-Young;Lee, Yong-Chang;Lee, Jong-Jin;Song, Young-Gak;Nam, Hyang-Mi
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.375-384
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    • 2007
  • This study was conducted to investigate the characteristics of brucellosis in Korean native cattle in a farm where bovine brucellosis was confirmed 3 times from September 2006 to March 2007. Of 74 bulls serum samples examined, 21 (28.4%) were positive by Rose-Bengal test (RBT) and Standard tube agglutination test (STAT). In the isolation test from seropositive bulls, B abortus was isolated and identified from 2 specimens (testis, intestinal lymph node) among 6 kinds of specimens including blood, urine, feces and soil. Isolation rate of intestinal lymph node and testis was 25% (3/12 cases) and 16.7% (2/12), respectively. B abortus was also isolated from calves below 12 months old, i.e., 1 isolate (25.0%) was confirmed from testis, 4 (40.0%) from supra-mammary lymph nodes and 1 (25.0%) from intestinal lymph node. All isolates had Brucella specific 16s r-RNA with 905-bp band detected by PCR assay. For the more effective control of bovine brucellosis in korea, this paper would like to suggest that all of bulls and calves should be included in the screening tests.

Studies on incidence of Bovine Brucellosis in Cheju-Do (제주도내 축우 부루셀라병 발생상황 조사)

  • 김우택;이완수;김공식
    • Korean Journal of Veterinary Service
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    • v.14 no.2
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    • pp.104-109
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    • 1991
  • Tile present study was carried out to investigate the incidence of bovine brucellsis in Cheju-do during the period from 1985 to 1990. The results were summarized as follows. 1. In the total 239,238 cattles tested. 1180(0.49%) were positive by standard tube agglutination test during the period from 1985 to 1990. 2. The major causes of incidence on brucellosis was grazing with carriers and repeated incidence in a herd. 3. The 13 Brucella abortus biotype 1 isolated from 10(50%) of 20 cattles slaughtered on brucellosis in 1990.

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STUDIES ON THE AGGLUTINATION REACTION OF BRUCELLA R-FORM BY ACRIFLAVIN AND HEAVY METAL IONS (Acriflavine 및 중금속이온반응에 의한 Brucella R형균 검출에 관한 연구)

  • Kang Byong-Kyu
    • Journal of the korean veterinary medical association
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    • v.10 no.1
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    • pp.13-23
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    • 1966
  • 1. To evaluate the quantitative relationship between S-R variation in 8r. abortus strain 19 and Br. melitensis 16 M induced by homologous immune serum treatment was studied by means of tube agglutination reaction with acriflavine add various heavy metal i

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The Bactericidal Effect of High Temperature Is an Essential Resistance Mechanism of Chicken Macrophage against Brucella abortus Infection

  • Arayan, Lauren Togonon;Reyes, Alisha Wehdnesday Bernardo;Hop, Huynh Tan;Xuan, Huy Tran;Baek, Eun Jin;Min, Wongi;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1837-1843
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    • 2017
  • Knowledge of avian host responses to brucellosis is critical to understanding how birds resist this infection; however, this mechanism is not well established. On the other hand, temperature has a major involvement in the physiology of living organisms, and cell death induced by heat is attributed to protein denaturation. This study demonstrates the direct bactericidal effect of a high temperature ($41^{\circ}C$) on Brucella abortus that resulted in the gradual reduction of intracellular bacteria and inhibited bacterial growth within avian macrophage HD11 in an increasing period of time. On the other hand, this study also revealed that high temperature does not affect the rate of bacterial uptake, as confirmed by the bacterial adherence assay. No significant difference was observed in the expression of target genes between infected and uninfected cells for both temperatures. This study suggests the susceptibility of B. abortus to bacterial death under a high temperature with an increased period of incubation, leading to suppression of bacterial growth.

Investigation of a Series of Brucellosis Cases in Gyeongsangbuk-do during 2003-2004 (2003-2004년 경상북도에서 발생한 브루셀라증)

  • Min, Young-Sun;Lee, Hwan-Seok;Lim, Hyun-Sul
    • Journal of Preventive Medicine and Public Health
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    • v.38 no.4
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    • pp.482-488
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    • 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.

Evaluation of Th1/Th2-Related Immune Response against Recombinant Proteins of Brucella abortus Infection in Mice

  • Im, Young Bin;Park, Woo Bin;Jung, Myunghwan;Kim, Suk;Yoo, Han Sang
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1132-1139
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    • 2016
  • Brucellosis is a zoonotic disease caused by Brucella, a genus of gram-negative bacteria. Cytokines have key roles in the activation of innate and acquired immunities. Despite several research attempts to reveal the immune responses, the mechanism of Brucella infection remains unclear. Therefore, immune responses were analyzed in mice immunized with nine recombinant proteins. Cytokine production profiles were analyzed in the RAW 264.7 cells and naive splenocytes after stimulation with three recombinant proteins, metal-dependent hydrolase (r0628), bacterioferritin (rBfr), and thiamine transporter substrate-binding protein (rTbpA). Immune responses were analyzed by ELISA and ELISpot assay after immunization with proteins in mice. The production levels of NO, TNF-α, and IL-6 were time-dependently increased after having been stimulated with proteins in the RAW 264.7 cells. In naive splenocytes, the production of IFN-γ and IL-2 was increased after stimulation with the proteins. It was concluded that two recombinant proteins, r0628 and rTbpA, showed strong immunogenicity that was induced with Th1-related cytokines IFN-γ, IL-2, and TNF-α more than Th2-related cytokines IL-6, IL-4, and IL-5 in vitro. Conversely, a humoral immune response was activated by increasing the number of antigen-secreting cells specifically. Furthermore, these could be candidate diagnosis antigens for better understanding of brucellosis.