OBJECTIVES: Brucellosis is a major zoonotic disease that poses a significant public health threat worldwide. The classical bacteriological detection process used to identify Brucella spp. is difficult and time-consuming. This study aimed to develop a novel molecular assay for detecting brucellosis. METHODS: All complete sequences of chromosome 1 with 2.1-Mbp lengths were compared among all available Brucella sequences. A unique repeat sequence (URS) locus on chromosome 1 could differentiate Brucella abortus from Brucella melitensis. A primer set was designed to flank the unique locus. A total of 136 lymph nodes and blood samples were evaluated and classified by the URS-polymerase chain reaction (PCR) method in 2013–2014. RESULTS: Biochemical tests and bacteriophage typing as the golden standard indicated that all Brucella spp. isolates were B. melitensis biovar 1 and B. abortus biovar 3. The PCR results were the same as the bacteriological method for detecting Brucella spp. The sensitivity and specificity of the URS-PCR method make it suitable for detecting B. abortus and B. melitensis. CONCLUSION: Quick detection of B. abortus and B. melitensis can provide the most effective strategies for control of these bacteria. The advantage of this method over other presented methods is that both B. abortus and B. melitensis are detectable in a single test tube. Furthermore, this method covered 100% of all B. melitensis and B. abortus biotypes. The development of this URS-PCR method is the first step toward the development of a novel kit for the molecular identification of B. abortus and B. melitensis.
Bacteria ; Bacteriophage Typing ; Brucella abortus* ; Brucella melitensis* ; Brucella* ; Brucellosis ; Chromosomes, Human, Pair 1 ; Lymph Nodes ; Methods ; Polymerase Chain Reaction* ; Public Health ; Sensitivity and Specificity ; Zoonoses

Brucellosis is a zoonotic infection that is usually transmitted from cattle to humans through ingestion of animal milk, direct contact with animal parts, or inhalation of aerosolized particles. In Korea, brucellosis seem to be transmitted through close contact with blood, fetus, urine, and placenta of domestic cow that has been infected by Brucella abortus, or inhalation of B. arbortus while examining or slaughtering cow. Brucella melitensis infection is rare in Korea and there have been no reported cases of B. melitensis originating from other countries until now. This report details a case of complicated brucellosis with infective spondylitis in a 48-year-old male construction worker recently returned from Iraq. Infection with B. melitensis was confirmed using 16s rRNA sequencing and omp31 gene analysis. The patient was successfully treated using a combination of rifampin, doxycycline, and streptomycin, in accordance with WHO guidelines. This is the first reported case of complicated brucellosis with infective spondylitis in Korea caused by B. melitensis originating from Iraq.
Animals ; Brucella abortus ; Brucella melitensis* ; Brucella* ; Brucellosis ; Cattle ; Doxycycline ; Eating ; Fetus ; Humans ; Inhalation ; Iraq ; Korea* ; Male ; Middle Aged ; Middle East ; Milk ; Placenta ; Rifampin ; Spondylitis ; Streptomycin ; Zoonoses

Brucellosis is a zoonotic infection that is usually transmitted from cattle to humans through ingestion of animal milk, direct contact with animal parts, or inhalation of aerosolized particles. In Korea, brucellosis seem to be transmitted through close contact with blood, fetus, urine, and placenta of domestic cow that has been infected by Brucella abortus, or inhalation of B. arbortus while examining or slaughtering cow. Brucella melitensis infection is rare in Korea and there have been no reported cases of B. melitensis originating from other countries until now. This report details a case of complicated brucellosis with infective spondylitis in a 48-year-old male construction worker recently returned from Iraq. Infection with B. melitensis was confirmed using 16s rRNA sequencing and omp31 gene analysis. The patient was successfully treated using a combination of rifampin, doxycycline, and streptomycin, in accordance with WHO guidelines. This is the first reported case of complicated brucellosis with infective spondylitis in Korea caused by B. melitensis originating from Iraq.
Animals ; Brucella abortus ; Brucella melitensis* ; Brucella* ; Brucellosis ; Cattle ; Doxycycline ; Eating ; Fetus ; Humans ; Inhalation ; Iraq ; Korea* ; Male ; Middle Aged ; Middle East ; Milk ; Placenta ; Rifampin ; Spondylitis ; Streptomycin ; Zoonoses

Brucella is an intracellular pathogen that invades a host and settles in its immune cells; however, the mechanism of its intracellular survival is unclear. Modification of small ubiquitin-related modifier (SUMO) occurs in many cellular activities. E2 conjugating enzyme 9 (Ubc9) is the only reported ubiquitin-conjugating enzyme that links the SUMO molecule with a target protein. Brucella's intracellular survival mechanism has not been studied with respect to SUMO-related proteins and Ubc9. Therefore, to investigate the relationship between Brucella melitensis 16M and SUMO, we constructed plasmids and cells lines suitable for overexpression and knockdown of SUMO1 and Ubc9 genes. Brucella 16M activated SUMO1/Ubc9 expression in a time-dependent manner, and Brucella 16M intracellular survival was inhibited by SUMO1/Ubc9 overexpression and promoted by SUMO1/Ubc9 depletion. In macrophages, Brucella 16M-dependent apoptosis and immune factors were induced by SUMO1/Ubc9 overexpression and restricted by SUMO1/Ubc9 depletion. We noted no effect on the expressions of SUMO1 and Ubc9 in B. melitensis 16M lipopolysaccharide-prestimulated mouse RAW264.7 macrophages. Additionally, intracellular survival of the 16M△VirB2 mutant was lower than that of Brucella 16M (p < 0.05). VirB2 can affect expression levels of Ubc9, thereby increasing intracellular survival of Brucella in macrophages at the late stage of infection. Collectively, our results demonstrate that B. melitensis 16M may use the VirB IV secretion system of Brucella to interact with SUMO-related proteins during infection of host cells, which interferes with SUMO function and promotes pathogen survival in host cells.
Animals ; Apoptosis ; Brucella melitensis ; Brucella ; Immunologic Factors ; Macrophages ; Mice ; Plasmids

Spinal epidural abscess, if especially caused by Brucellosis is a very rare disease which is usually a consequence of spondylodiscitis. The spinal column can be affected at any joint; however, the lumbar spine is the most common region, especially at the level of the L4-5 and L5-S1. The frequency of spinal involvement usually seen at the lumbar, thoracic and cervical spine respectively. As an occupational disease in farmers, veterinaries, butchers, laboratory staff and shepherds, brucellosis can also occur by direct contact to animals and infected materials or ingestion of raw cheese, milk or unpasteurized milk products. In this study, we presented two cases with cervical spinal epidural abscess caused by brucella melitensis, which was successfully treated by surgical approach. Initial treatment was combined with antibiotic therapy after the surgery for 3 months.
Animals ; Brucella ; Brucella melitensis ; Brucellosis ; Cheese ; Discitis ; Eating ; Epidural Abscess ; Milk ; Occupational Diseases ; Rare Diseases ; Spinal Cord Compression ; Spine

No Abstract Available.
Brucella abortus* ; Brucella* ; Immunity, Humoral* ; Jeju-do* ; Recombinant Proteins*

A real-time PCR assay using hybridization probe (HybProbe) has been developed to detect Brucella (B.) melitensis strains. The primer and HybProbe sets were designed based on the gap gene of chromosome I with a specific single nucleotide polymorphism of B. melitensis. Specificity of the assay was confirmed by comparison to reference Brucella species and other related strains. In the melting curve analysis, B. melitensis generated a peak at 67degrees C unlike those for other Brucella species observed at 61degrees C. Sensitivity of the assay for B. melitensis ranged from 20 ng to 200 fg of genomic DNA. The ability to identify 94 Mongolian B. melitensis isolates using the real-time PCR assay was identical to that of classical biotyping methods and differential multiplex PCR. These data showed that this new molecular technique is a simple and quick method for detecting B. melitensis, which will be important for the control and prevention of brucellosis.
Brucella ; Brucella melitensis* ; Brucellosis ; DNA ; Freezing ; Mongolia* ; Multiplex Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Real-Time Polymerase Chain Reaction* ; Sensitivity and Specificity

The occurrence of Brucella endocarditis following Bentall operation is a rare and life threatening condition, particularly when it is complicated by development of a pseudoaneurysm. Here we present a 40-year-old veterinarian with bicuspid aortic valve, who developed type A aortic root dissection and required Bentall operation. His past medical history was positive for an episode of treated brucellosis. Four months after the operation, he developed signs, symptoms and serological findings of brucellosis for the second time and was treated with antibiotics. Then after he was well untill about 14 days prior to his recent admission, when he again developed hip pain, fever, shortness of breath, profound fatigue and weakness. His transthoracic echocardiography was suggestive of endocarditis. A transesophageal echocardiogram revealed detachment of valve-conduit from the annulus and the mitral-aortic intervalvular fibrosa and the presence of a large aortic pseudoaneurysm with multiple vegetations attached to its Dacron walls. A huge pseudoaneurysm was detected at surgery and the whole valve-conduit was replaced with a 25 mm homograft. Blood and vegetation cultures turned to be positive for Brucella melitensis. Early echocardiographic diagnosis and prompt surgical intervention were helpful for survival of our patient who had aortic pseudoaneurysm complicated by Brucella endocarditis.
Adult ; Aneurysm, False* ; Anti-Bacterial Agents ; Aortic Valve ; Bicuspid ; Brucella melitensis ; Brucella* ; Brucellosis ; Diagnosis ; Dyspnea ; Echocardiography ; Endocarditis* ; Fatigue ; Fever ; Hip ; Humans ; Polyethylene Terephthalates ; Transplantation, Homologous ; Transplants* ; Veterinarians

Brucella (B.) canis is mainly transmitted by direct or indirect contact with aborted fetuses and placenta. It's also known to be able to infect human, which likely results in providing veterinarians and companion animal owners for infectious risk. To develop diagnostic ELISA, we cloned and expressed rp1L gene of B. canis, which encodes the ribosomal protein L7/L12. Using this purified recombinant protein, indirect-ELISA (iELISA) was evaluated using 78 positive and 44 negative sera. The sensitivity and the specificity of iELISA were 94% and 89%, respectively. The results indicated that indirect-ELISA using recombinant ribosomal protein L7/L12 may be useful for diagnosis of canine brucellosis.
Aborted Fetus ; Brucella ; Brucella canis ; Brucellosis ; Clone Cells ; Enzyme-Linked Immunosorbent Assay ; Humans ; Pets ; Placenta ; Ribosomal Proteins ; Sensitivity and Specificity ; Veterinarians

OBJECTIVETo understand the genetic and epidemiologic characteristic of Brucella (B.) melitensis strains isolated in Guizhou province in 2010-2012.

METHODSB. genus specific BCSP31-PCR and species-specific AMOS-PCR were used to identify the bacteria strain, while the identified strains were analyzed under MLVA-16 and cluster analysis of B. melitensis strains. The strains were isolated from Guizhou and other provinces.

RESULTSSix B. melitensis strains were identified as B. melitensis using the BCSP31-PCR and AMOS-PCR. Data from the MLVA-16 analysis revealed the differences of repeated numbers at parts of the VNTR locus in the six strains isolated in Guizhou province. The six strains from Guizhou province and 105 B. melitensis strains from other province could be divided into 72 MLVA types(MT). Strain ZY and ZA from Guizhou province were typed as MT63, and LL3, LL4 and LL11 were typed as MT67, while strain SQ was typed as MT72. Data from the clustering analysis showed that ZY,ZA, LL3, LL4 and LL9 were most closely clustered with B. melitensis isolates from Yunnan, Fujian and Guangdong provinces, but strain SQ was genetically remote from other strains.

CONCLUSIONPCR methods, combined with MLVA-16, identified the six B. melitensis strains isolated in Guizhou province in 2010-2012 as B. melitensis biovar 3, with the genetic diversity of the strains showed. Six strains were closely related to the B. melitensis strains from Yunnan, Fujian and Guangdong provinces. The results of this study provided scientific basis for the control and prevention of Brucellosis in Guizhou province.

Brucella melitensis ; genetics ; isolation & purification ; Brucellosis ; epidemiology ; China ; epidemiology ; Cluster Analysis ; Genetic Variation ; Humans ; Minisatellite Repeats ; Polymerase Chain Reaction