• Title/Summary/Keyword: Brucella spp.

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Detection of Brucella spp. and Leptospira interrogans in the Canine Blood by Multiplex Nested PCR (개 혈액에서 Multiplex Nested PCR기법을 이용한 Brucella spp. 및 Leptospira interrogans 검출)

  • Lee, Jung-Youn;Lee, Sang-Eun;Kim, Suk;Kim, Duck-Hwan;Song, Kun-Ho
    • Journal of Veterinary Clinics
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    • v.25 no.4
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    • pp.241-244
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    • 2008
  • This study examined the prevalence of Brucella spp. and Leptospira interrogans in 360 clinically healthy dogs using multiplex nested PCR. Four dogs (1.1%, 2 females and 2 males) tested positive to Brucella spp. by multiplex nested PCR. Fifty nine (16.4%, 31 females and 28 males) of 360 dogs tested positive L. interrogans. In 1 and 2 of the samples that tested positive to Brucella spp. and L. interrogans, the partial sequences of the virB1 and 16S rRNA genes were identified by direct sequence analysis, respectively. In conclusion, prevalence of Brucella spp. and L. interrogans by multiplex nested PCR revealed low and high, respectively. Multiplex nested PCR is can be useful for early detection of Brucella spp. and L. interrogans in the canine blood from asymptomatic dogs.

Epidemiological study for infection route of brucellosis in a infected dairy farms (Brucella 감영농장에서 감염경로의 역학적 연구)

  • 윤여백;김영진;김추철;노영선;권미순;김철민;임채웅
    • Korean Journal of Veterinary Service
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    • v.27 no.2
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    • pp.159-164
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    • 2004
  • A dairy farm that has been suffered continuously(more than 2 years) from brucellosis in Korea in spite of repeated legal test-and-slaughter was investigated the main source of infection in the farm. All cattle(22 milking cows, 44 heifers, 60 calves, 8 bull), dogs(3 mixed breed), feces from wild birds(3 samples), drinking water(3 sites), and soil in the paddocks(14 sites) inside the farm were examined with serological and/or bacteriological methods including specific DNA detection with PCR method. Brucella spp in the milk and blood were detected in 12/22 and 5/22 milking cows, respectively, although all of them were negative with conventional tube agglutination test. The number of serologically positive heifer was 15(15/44), but the isolation of Brucella spp was succeeded in the only 11(11/15) of them. Brucella were detected in vagina 1(1/11) and nasal(3/12) excretion in serologically positive heifers. All the three dogs were serologically positive, and Brucella spp were isolated from their blood. However, Brucella spp were not detected in the drinking water, soil in the paddocks, nor the feces of wild birds. The results suggest that milking cow secrete Brucella spp through milk, genital tract and nasal cavity, which are the major source of infection in this farm, The main infection route of Brucella spp is contact to contact with Brucella spp excreting animals rather than environmental contamination. The animals, living together with infected cow such as dogs, are the readily susceptible and are required to be examined for Brucella spp.

Occurrence of canine brucellosis in large kennels and characterization of Brucella canis isolates by PCR-RFLP (집단 개사육농장에서의 Canine Brucellosis 발생 및 PCR-RFLP를 이용한 분리주의 특성조사)

  • Kim, Jong-Wan;Lee, Young-Ju;Tak, Ryun-Bin
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.67-75
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    • 2003
  • A total of 260 dogs were randomly selected from two different treed kennels that brucellosis has occurred (group 1, 126 dogs), and random selected breed kennel (group 2, 134 dogs), and monitored for Brucella canis (B. canis) by 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and bacterial culture method. For the differentiation, PCR-RFLP using omp-31, wbkA and per genes used for 52 of B canis strains (strain I) isolated in this study and 3 of B. canis strains (strain II) isolated in 1994 in Korea. 2ME-RSAT revealed that 63/126 dogs (50.0%) and 12/134 dogs (9.0%) were positive in group I and group II, respectively. Bacterial culture revealed that 47/126 dogs (37.3%) and 5/134 dogs (3.7%) were positive in group I and group II, respectively. As the results of PCR-RFLP, $\underline{omp}-31$ was amplified from all Brucella spp, except B. abortus. All B. canis isolates showed unique PCR-RFLP pattern following digestion with Bmel8I. However, all Brucella spp. showed the same PCR-RFLP pattern following digestion with SalI. PCR-RFLP analysis of wbkA revealed that all Brucella spp. showed the same pattern following digestion with HindIII. PCR-RFLP analysis of per revealed that B. abortus 544 and B. melitensis 63/9 showed the same pattern, but different from B. suis and B. canis following digestion with HindIII.

Biological characterization of Brucella spp. isolated from cattle in Gyeongbuk, Korea (국내 경북지역 소에서 분리된 브루셀라 분리주의 생물학적 특성)

  • Kim, Jeong-Hwa;Lim, Jeong Ju;Kim, Dong Hyeok;Lee, Jin Ju;Kim, Dae Geun;Jun, Moo-Hyung;Kim, Sang Hun;Chang, Hong Hee;Lee, Hu Jang;Min, Won-Gi;Kim, Suk
    • Korean Journal of Veterinary Research
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    • v.50 no.2
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    • pp.117-124
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    • 2010
  • Members of the genus Brucella are facultative intracellular bacteria and cause brucellosis, a chronic disease in humans and abortion in animals. In this study, we tested sera for brucellosis of 15 Hanwoo farms in the western part of Gyeong-buk province, resulting 5 farms were brucellosis positive in 2008. We collected blood from 277 heads in the brucellosis positive 5 farms, and performed serological diagnosis, brucella positive cattle which had shown higher than 200 antibody titer in tube agglutination test were slaughtered, supramammary lymph nodes were collected, and Brucella spp. wild type isolation and identification were performed. From these results, 15 of Brucella spp. wild type strains were isolated and all strains were identified as B. abortus biotype 1 by biological and molecular analysis. In the antimicrobial susceptibility test, all 15 strains had a similar susceptibility and resistance pattern. This study may be useful for bacteriological and epidemiological understanding of cattle brucellosis in Korea.

Antimicrobial efficacies of alkaline disinfectant solution and commercial disinfectants against Brucella ovis

  • Yoo, Jong-Hyun
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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    • pp.347-351
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    • 2009
  • Bruella spp. involves a considerable danger of public health and farm animal industry. In this study, we assessed the disinfection efficacy of alkaline disinfectant solution and three commercial farm disinfectants (quaternary ammonium compound, sodium dichloroisocyanurate, potassium monopersulphate/sodium dichloroisocyanurate) against Brucella ovis. A bactericidal efficacy test by broth dilution method was used to determine the lowest effective dilution of selected disinfectants following exposure to test bacteria for 30 minutes at $4^{\circ}C$. Disinfectants and test bacteria are diluted with distilled water (DW), hard water (HW) or organic matter suspension (OM) according to treatment condition. Three commercial disinfectant showed excellent antimicrobial activity (up to dilution of $\times200$ in OM treatment). Alkaline disinfectant solution demonstrated favorable bactericidal efficacy against B. abortus (at dilution of $\times20$ in OM treatment). Three commercial farm disinfectants possess excellent efficacy against B. ovis. Alkaline disinfectant solution has lower potency than commercial farm disinfectant but could help to limit the spread of brucellosis.

Brucellosis outbreak of Korean indigenous cattle at Yeongwol and Pyeongchang county in Korea

  • Kim Song-Tae;Yoon Kyo-Bok;Kang Tae-Kyu;Bak Won-Hern;Lee Jung-Hyun;Chung Dong-Su
    • Korean Journal of Veterinary Service
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    • v.28 no.4
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    • pp.387-392
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    • 2005
  • This study was attempted to investigate the properties of brucellosis in Korean indigenous cattle at the Yeongwol and Pyeongchang county. Brucella spp was differentiated and identified from cotyledons, amniotic fluids and supramammary lymph nodes which confirmed with clinical, serological, epidemiological evidences (69 cases) from January to June, 2004. Isolation frequency of this causative agent from supramammary lymph nodes, cotyledons and amniotic fluids from 38 pregnant Korean indigenous cattle were $39.1\%,\;87.5\%,\;and\;63.2\%$, respectively, and finally confirmed with Brucella abortus biotype 1 through biochemical and serological test. A Brucella specific DNA with 711bp band was detected by PCR assay using BCSP primer. The two cases were definite epidemiological evidences that infected Korean indigenous cattle acrossed the border to Yeongwol and Pyeongchang from near two provinces. Effective prevention programs are urgently needed for further spreading this epidemics.

A New ELISA Kit Based on Antigenic Epitopes for Diagnosing Brucella abortus

  • Khodabakhsh, Tannaz;Arabi, Azin;Pakzad, Parviz;Gheflat, ShivaSadat;Bahreinipour, Akram;Bandehpour, Mojgan
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.158-163
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    • 2019
  • Brucellosis is one of the most important zoonotic diseases that lead to a great amount of economic losses. Prevention and diagnosis are both necessary to eradicate this disease. The identification and evaluation of different antigens of Brucella spp. play a key role in the progress of diagnostic programs. In this study, we designed, produced, and evaluated a 24-kDa polypeptide containing antigenic epitopes of VirB2, 3, and 9 of Brucella abortus for use with the ELISA kit. The produced polypeptide is appropriate for diagnosing brucellosis in bovines by a laboratory diagnostic kit, with 100% sensitivity and 97.5% specificity.

Bactericidal Efficacy of Vital-Oxide®, Disinfectant Solution Against Salmonella Typhimurium and Brucella Ovis

  • Cha, Chun-Nam;Lee, Yeo-Eun;Kang, In-Jin;Yoo, Chang-Yeul;An, Sun-Jeong;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.50-54
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    • 2012
  • $Salmonella$ spp. and $Brucella$ spp. have caused a considerable disease of farmed animals and economic loss in animal farming and food industry. In this study, the disinfection efficacy of Vital-$Oxidel^{(R)}$, a commercial disinfectant, composed to chlorine dioxide, betaine hydrochloride, and propylene glycol was evaluated against $S.$ $typhimurium$ and $Brucella$ $ovis$. A bactericidal efficacy test by broth dilution method was used to determine the lowest effective dilution of the disinfectant following exposure to test bacteria for 30 min at $4^{\circ}C$. Vital-$Oixdel^{(R)}$ and test bacteria were diluted with distilled water (DW), hard water (HW) or organic matter suspension (OM) according to treatment condition. On OM condition, the bactericidal activity of Vital-$Oixdel^{(R)}$ against S. typhimurium and Brucella ovis was lowered compared to that on HW condition. As Vital-$Oxidel^{(R)}$ possesses bactericidal efficacy against animal pathogenic bacteria such as $S.$ $typhimurium$ and $Brucella$ $ovis$, this disinfectant solution can be used to control the spread of bacterial diseases.

A Brucella Omp16 Conditional Deletion Strain Is Attenuated in BALB/c Mice

  • Zhi, Feijie;Fang, Jiaoyang;Zheng, Weifang;Li, Junmei;Zhang, Guangdong;Zhou, Dong;Jin, Yaping;Wang, Aihua
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.6-14
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    • 2022
  • Brucella spp. are facultative intracellular pathogens that invade, survive and proliferate in numerous phagocytic and non-phagocytic cell types, thereby leading to human and animal brucellosis. Outer membrane proteins (Omps) are major immunogenic and protective antigens that are implicated in Brucella virulence. A strain deleted of the omp16 gene has not been obtained which suggests that the Omp16 protein is vital for Brucella survival. Nevertheless, we previously constructed an omp16 conditional deletion strain of Brucella, ∆Omp16. Here, the virulence and immune response elicted by this strain were assessed in a mouse model of infection. Splenomegaly was significantly reduced at two weeks post-infection in ∆Omp16-infected mice compared to infection with the parental strain. The bacterial load in the spleen also was significantly decreased at this post-infection time point in ∆Omp16-infected mice. Histopathological changes in the spleen were observed via hematoxylin-eosin staining and microscopic examination which showed that infection with the ∆Omp16 strain alleviated spleen histopathological alterations compared to mice infected with the parental strain. Moreover, the levels of humoral and cellular immunity were similar in both ∆Omp16-infected mice and parental strain-infected mice. The results overall show that the virulence of ∆Omp16 is attenuated markedly, but that the immune responses mediated by the deletion and parental strains in mice are indistinguishable. The data provide important insights that illuminate the pathogenic strategies adopted by Brucella.

Biochemical characteristics of Brucella abortus isolated from cattle in Gyungbuk province (경북지방 소에서 분리한 Brucella abortus의 생화학적 특성)

  • Kim, Seong-Guk;Kim, Young-Hoan;Cho, Min-Hee;Lee, Young-Ju;Park, Cheong-Kyu
    • Korean Journal of Veterinary Service
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    • v.32 no.2
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    • pp.139-146
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    • 2009
  • Bovine brucellosis is a zoonosis, long incubation period and chronic infectious disease, usually caused by Brucella abortus. This study was carried out to investigate the biotyping and biochemical characterization of B. abortus isolated from 208 farm 871 korean cattle and holstein diagnosed brucellosis by serological positive in Gyeongbuk province during the period from 2002 to 2006. B. abortus was isolated from 124 (14.2%) of 871 cattle, and isolated 110 (13.4%) of 820 Korean cattle and 14 (27.5%) of 51 holstein in breed. The uterus of korean cattle was isolated in 8 (17.8%) of 45 cattle and supramammary lymph none of holstein was isolated 11 (68.8%) of 16 cattle. 101 (12.5%) of 810 serological positive blood samples were isolated B. abortus. The isolation rate of B. abortus was correlated with antibody titers. The biochemical characterization of isolates was non-hemolytic, production of H$_2$S, oxidase-positive, catalase-positive, hydrolyzation of urea and growth of basic fuchsin dye medium. As a result, all of isolates was identified B. abortus bv 1. 124 isolates were susceptible to ampicillin, lincospectin, amikacin, gentamicin, kanamycin, neomycin, streptomycin, tetracycline, ciprofloxacin, norfloxacin and enrofloxacin.