• Title/Summary/Keyword: Brucella

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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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Intracellular Trafficking Modulation by Ginsenoside Rg3 Inhibits Brucella abortus Uptake and Intracellular Survival within RAW 264.7 Cells

  • Huy, Tran Xuan Ngoc;Reyes, Alisha Wehdnesday Bernardo;Hop, Huynh Tan;Arayan, Lauren Togonon;Min, WonGi;Lee, Hu Jang;Rhee, Man Hee;Chang, Hong Hee;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.616-623
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    • 2017
  • Ginsenoside Rg3, a saponin extracted from ginseng, has various pharmacological and biological activities; however, its effects against Brucella infection are still unclear. Herein, the inhibitory effects of ginsenoside Rg3 against intracellular parasitic Brucella infection were evaluated through bacterial infection, adherence assays, and LAMP-1 colocalization, as well as immunoblotting and FACS for detecting MAPK signaling proteins and F-actin polymerization, respectively. The internalization, intracellular growth, and adherence of Brucella abortus in Rg3-treated RAW 264.7 cells were significantly decreased compared with the Rg3-untreated control. Furthermore, an apparent reduction of F-actin content and intensity of F-actin fluorescence in Rg3-treated cells was observed compared with B. abortus-infected cells without treatment by flow cytometry analysis and confocal microscopy, respectively. In addition, treating cells with Rg3 decreased the phosphorylation of MAPK signaling proteins such as ERK 1/2 and p38 compared with untreated cells. Moreover, the colocalization of B. abortus-containing phagosomes with LAMP-1 was markedly increased in Rg3-treated cells. These findings suggest that ginsenoside Rg3 inhibits B. abortus infection in mammalian cells and can be used as an alternative approach in the treatment of brucellosis.

Brucella melitensis omp31 Mutant Is Attenuated and Confers Protection Against Virulent Brucella melitensis Challenge in BALB/c Mice

  • Verdiguel-Fernandez, L;Oropeza-Navarro, R;Ortiz, Adolfo;Robles-Pesina, MG;Ramirez-Lezama, J;Castaneda-Ramirez, A;Verdugo-Rodriguez, A
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.497-504
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    • 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.

Comparison of diagnostic methods for detection of Brucella species in dog blood samples (개 혈액 재료에서의 Brucella 검출을 위한 진단방법의 비교)

  • Kwon, Soon-Oh;Lam, Truong Quang;Her, Moon;Ahn, Dong-Chun;Park, Sang-Hee;Park, Mi-Yeoun;Lee, Young-Ju;Hahn, Tae-Wook
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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    • pp.335-341
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    • 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.

The virulence of Brucella abortus isolated from cattle in Korea (국내 소에서 분리한 Brucella abortus의 병원성 분석)

  • Lim, Jeong Ju;Kim, Jeong-Hwa;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.51 no.1
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    • pp.15-20
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    • 2011
  • In this study, we isolated 12 of Brucella (B.) spp. from cattle, which have been positive in Rose Bangal test and tube agglutination test in Gyeongbuk province in 2009. According to AMOS PCR analysis, isolated 12 strains were identified as B. abortus. Murine derived macrophage, RAW 264.7 cells, were infected with isolated 12 strains or reference strain (B. abortus 544), and bacterial internalization were characterized. According to these results, we divided the isolated strains into the following three groups: class I, lower internalization than that of B. abortus 544; class II, similar internalization to that of that of B. abortus 544; class III, higher internalization than that of B. abortus 544 within RAW 264.7 cells. Furthermore, intracellular growth, bacterial adherent assay, LAMP-1 colocalization, virulence in mice and surface protein pattern were characterized. From these results, representative strains of class III showed lower LAMP-1 colocalization, higher adherent efficiency, higher virulence in mice than those of B. abortus 544, and showed different pattern of surface proteins. These results suggest that B. abortus field strains, isolated from cattle in Korea, possess various virulence properties and higher internalization ability of field strain may have an important role for its virulence expression.

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.

A Genetic Comparison of Brucella abortus Isolates from Animals and Humans by Using the MLVA Assay

  • Her, Moon;Kang, Sung-Il;Kim, Jong-Wan;Kim, Ji-Yeon;Hwang, In-Yeong;Jung, Suk-Chan;Park, Sang-Hee;Park, Mi-Yeoun;Yoo, Han-Sang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1750-1755
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    • 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.

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.

Structure and Function of the Autolysin SagA in the Type IV Secretion System of Brucella abortus

  • Hyun, Yongseong;Baek, Yeongjin;Lee, Chanyoung;Ki, Nayeon;Ahn, Jinsook;Ryu, Sangryeol;Ha, Nam-Chul
    • Molecules and Cells
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    • v.44 no.7
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    • pp.517-528
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    • 2021
  • A recent genetic study with Brucella abortus revealed the secretion activator gene A (SagA) as an autolysin component creating pores in the peptidoglycan (PGN) layer for the type IV secretion system (T4SS) and peptidoglycan hydrolase inhibitor A (PhiA) as an inhibitor of SagA. In this study, we determined the crystal structures of both SagA and PhiA. Notably, the SagA structure contained a PGN fragment in a space between the N- and C-terminal domains, showing the substrate-dependent hinge motion of the domains. The purified SagA fully hydrolyzed the meso-diaminopimelic acid (DAP)-type PGN, showing a higher activity than hen egg-white lysozyme. The PhiA protein exhibiting tetrameric assembly failed to inhibit SagA activity in our experiments. Our findings provide implications for the molecular basis of the SagA-PhiA system of B. abortus. The development of inhibitors of SagA would further contribute to controlling brucellosis by attenuating the function of T4SS, the major virulence factor of Brucella.