• 대한수의학회지
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• 제49권2호
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• pp.105-111
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• 2009

In this study, occurrence of canine brucellosis was surveyed in kennels, indoor dogs and stray dogs in Korea, and infrequent restriction site-polymerase chain reaction (IRS-PCR) was applied to analyze DNA polymorphism of Brucella canis (B. canis) isolates. Among a total of 501 dogs tested, B. canis antibodies by both rapid screening agglutination with 2-mercaptoethanol (2-ME RSAT) and immunochromatographic assay were detected in only 14.1% of kennel dogs. There were no seropositive cases in indoor dogs and stray dogs. DNA polymorphism was observed in 16 B. canis isolates by the IRS-PCR. Sixteen isolates were tested with primers, PsalA, PsalC, PsalG and PsalT, and different primers produced different DNA patterns. In regard to the IRS-PCR pattern of 16 isolates, 9 (56.3%) belonged to the IRS-PCR type I. The remaining 7 were differentiated as type II, III and IV. An application of the primer PsalC provided discrimination between B. canis isolated in 2005 and others.

• 대한수의학회지
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• 제45권2호
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• pp.199-205
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• 2005

DNA fingerprint patterns of 8 Brucella reference strains and 15 B. abortus field isolates were characterized by repetitive element sequence-based PCR (rep-PCR) using BOX- and ERIC-primers in this study. AMOS PCR differentiated all Brucella field isolates from B. abortus RB51, a vaccine strain by producing a B. abortus-specific 498 bp band. Rep-PCR using BOX-primer produced 13 to 18 bands with sizes of between 230 and 3,300 bp, and discriminated Brucella strains to the species level except B. canis and B. suis. PCR products amplified with ERIC primers were, however, not appropriate for differentiating the Brucella isolates. DNA fingerprint patterns for all B. abortus field isolates were identical among them and were put on one cluster with B. abortus biovar 1 reference strain in the dendrogram, indicating they were highly clonal. These results suggested that rep-PCR using BOX primer might to be a useful tool for calculating genetic relatedness among the Brucella species and for the study of brucellosis epidemiology.

• 한국동물위생학회지
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• 제28권4호
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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.

• 대한수의학회지
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• 제9권2호
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• pp.55-59
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• 1969

For the establishment of simpler and more reliable methods for the diagnosis of Brucellosis, diagnostic antigen for Milk Ring Test (M.R.T.) was prepared according to working document recommended by Ministry of Agriculture U.S.A., and applied in the field for the test of its's applicability. The results are summarized as follows. 1. Of fifty two dairy cattle ranches in the vicinity of Seoul, five ranges were shown positive reaction to M.R.T. 2. Even though one volume of positive milk was mixed with five to ten volumes of negative milk, the positive reactions were also observed. 3. Antigenicity of the antigen was maintained without deteriolation for 13 months at $5^{\circ}C$ and three months at both $22^{\circ}C$ and $37^{\circ}C$. 4. Three positive cows to M.R.T. were also shown suspective reaction by Serum Agglutination Test.

• 한국미생물·생명공학회지
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• 제47권1호
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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.

• 한국동물위생학회지
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• 제30권4호
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• pp.511-525
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• 2007

A seroprevalence study of small ruminant brucellosis was conducted in sheep and goat rearing selected areas of Mymensingh district and Dhaka district, Bangladesh, from March, 2005 to May, 2006. Sera from 62 sheep and 300 goats were tested by rose bengal plate test (RBPT), plate agglutination test (PAT), tube agglutination test (TAT) and mercaptoethanol test (MET). Out of the 62 sera tested 3.25% (n = 2) were positive to RBT, PAT and TAT and 4.84% (n = 3) were positive MET. In case of 300 goats, 1.67% (n = 5) were positive to RBT and PAT, 2% (n = 6) were positive to TAT and 2.33% (n = 7) were positive to MET. This investigation is the first of its type to be performed in small ruminants in Bangladesh. Higher prevalence rate (8.0 %) was found in BAU nutrition farm in case of sheep and 10 % in Bangladesh Agricultural University (BAU) Veterinary Clinic in case of goat while lower prevalence (0.0 %) was recorded in Pharmacology project and BAU adjacent villages in case of sheep and (0.0 %) in Dhamrai upazila in case of goats respectively. Brucella antibodies were more prevalent in sheep (8.84 %) than in goat (2.33 %).

• Asian-Australasian Journal of Animal Sciences
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• 제24권7호
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• pp.898-904
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• 2011

The PCR- restriction fragment length polymorphism (RFLP) in and around TM4 of SLC11A1 gene and its association with the incidences of brucellosis in Hariana breed (Bos indicus) and Holstein Friesian crossbred (Bos indicus${\times}$Bos taurus) cattle was examined. A fragment of 954 bp encoding the TM4 was amplified, and RFLP was identified by digestion of the amplicon independently with AluI and TaqI. The amplicon (GenBank Acc. No. AY338470 and AY338471) comprised of a part of exon V (<59 bp) and VII (62>), and entire intron 5 (423 bp), exon VI (71 bp) and intron 6 (339 bp). Digestion with AluI revealed the presence of two alleles viz, A (281, 255, 79 and 51 bp) and B (541, 255, 79 and 51 bp). The frequency of A allele was estimated as 0.80 and 0.73 in Hariana and crossbred cattle, respectively. Due to presence of a polymorphic TaqI site at intron 5, two alleles: T (552 and 402 bp) and Q (231, 321 and 402 bp) were identified. The frequency of T allele was estimated as 0.96 and 0.97, respectively. For association study, on the basis of serological tests and history of abortion, the animals were grouped into "affected" and "non-affected". However, no association could be established with the observed RFLPs.

• Journal of Microbiology and Biotechnology
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• 제30권4호
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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.

• 대한수의학회지
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• 제43권1호
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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.

• 대한수의학회지
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• 제56권3호
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• pp.147-153
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• 2016

A total of 782 blood and 465 tissue samples from 1,039 wild animals and 127 dairy goats were collected from January 2011 to December 2013 in 10 provinces of South Korea and tested for the presence of brucellosis. The Rose Bengal test revealed that 8.0% (52/650) of the serum samples were seropositive, while 4.2% (33/782) of the serum samples were positive for Brucella antibodies by competitive enzyme-linked immunosorbent assay. Of the 650 sera examined, only 16 (2.5%) were positive by both serological tests. Direct polymerase chain reaction (PCR) assay using B4/B5 primers for Brucella abortus (BCSP31) revealed the prevalence of Brucella to be 26.5% (129/487) in blood samples and 21% (98/465) in tissue samples while, 16S rRNA PCR detected Brucella DNA in 6.8% (33/487) and 2.6% (12/465) in blood and tissue samples, respectively. Of PCR-positive samples, only 6.2% (30/487) of blood samples and 2.4% (11/465) of tissue samples were found to be positive by both BCSP31 and 16S rRNA PCRs. However, Brucella strains were isolated by blood culture from only two out of 487 blood samples (0.4%). This characterization and identification of pathogenic Brucella isolates is the first to clearly indicate that the organisms were Brucella abortus biovar 1.