Endocarditis de la válvula aórtica causada por Brucella melitensis / Aortic valve endocarditis caused by Brucella melitensis

La endocarditis bacteriana secundaria a la infección por Brucella spp., en este caso B. melitensis, como complicación de la brucelosis humana tiene una incidencia baja y, aunque es la presentación clínica con la que se asocia más frecuentemente la mortalidad, no todos los casos son letales, si son tratados oportunamente. Se describe el caso clínico de una endocarditis bacteriana por B. melitensis, diagnosticada en un adulto por el aislamiento del microorganismo en el hemocultivo. Paciente del sexo masculino, de 40 años, con antecedentes de realizar partos en el ganado bovino y consumir leche no pasteurizada. Acudió al médico por presentar durante siete días de evolución de las siguientes manifestaciones clínicas: fiebre, mialgias, artralgias, tos seca y pérdida de peso (15 kg). El hemograma informa: leucopenia, trombocitopenia y anemia; mientras que en un ecocardiograma transesofágico se observó vegetación en la válvula aórtica con una disminución de la función sistólica y en el hemocultivo se aisló B. melitensis. Debido a estos antecedentes, se inició el tratamiento antibacteriano con rifampicina, doxiciclina y gentamicina. El paciente se recuperó y tuvo una evolución clínica satisfactoria. La brucelosis es una enfermedad infrecuente. Debe considerarse en toda persona con fiebre de foco desconocido que resida en zonas endémicas o esté expuesto al cuidado de animales de granja. En esta enfermedad se impone un diagnóstico y tratamiento preciso, por ser una complicación con alta letalidad.

Bacterial endocarditis, secondary to Brucella spp. infection, in this case by B. melitensis, as a complication of human brucellosis has a low incidence. Although it is the clinical presentation most frequently associated with mortality, not all cases are lethal if timely treatment is provided. We describe a clinical case of bacterial endocarditis due to B. melitensis in a 40-year-old male patient with a history of conducting cattle deliveries and consuming unpasteurized milk, diagnosed after isolating the microorganism in blood culture. He presented with the following clinical manifestations after seven days of evolution: fever, myalgias, arthralgias, dry cough and weight loss (15 kg). The hemogram revealed leukopenia, thrombocytopenia, and anemia; while a transesophageal echocardiogram showed vegetation on the aortic valve with decreased systolic function, and B. melitensis was isolated in a blood culture. Considering this medical history, antibacterial treatment was initiated with rifampicin, doxycycline and gentamicin. The patient recovered and had satisfactory clinical evolution. Brucellosis is a rare disease. It should be considered in any person with a fever of unknown origin who lives in endemic areas or is exposed to the care of farm animals. Endocarditis is a highly lethal complication of human brucellosis; therefore, it requires a precise diagnosis and treatment.

Determination of investigation of the link between human and animal Brucella isolates in Iran using multiple-locus variable number tandem repeat method comprising 16 loci (MLVA-16)

ABSTRACT Background: Epidemiological studies are important tools to assess the diversity of Brucella isolates and to estimate their epidemiological relationship among isolates from different geographical origins. In this study the MLVA16 (multiple-locus variable number tandem repeat analysis based on 16 loci) was employed to investigate the diversity of Brucella spp. Isolated from humans and animals for epidemiological purposes and to determine the most common Brucella genotypes in Iran. Methods: We designed a molecular-based study to evaluate the potential reservoirs of human brucellosis. After isolation and identification of 54 Brucella spp human and animal specimens from three regions of Iran, bacterial genomic DNA was extracted MLVA with three panel was used for the genotyping of isolates. The size of PCR products were analyzed and converted to repeat unit numbers using a published allele numbering system and data set was imported into Bionumerics. Results: Three isolates (5.55%) were identified as Brucella abortus and 51 (94.44%) as Brucella melitensis. Two isolates of Brucella abortus were from humans and one from an animal. Thirty-four Brucella melitensis isolates were from humans and 17 from animals. Using MLVA16-genotyping, 54 isolates with genetic similarity coefficient of 80% were divided into 46 genotypes and 22 genotypes were represented by a single isolate, while 4, 2, 1 and 2 genotypes were represented by 2, 3, 4 and 7 isolates, respectively. The most prevalent genotype was represented by 14 isolates. There were two other frequent genotypes each represented by seven isolates, among which only one was restricted to a geographic region. Discriminatory power for each locus was determined in this study and panel 2B shows the high discretionary power [Bruce04 (0.837), Bruce30 (0.806), Bruce 09 (0.787), Bruce 07 (0.772), Bruce16 (0.766)]. Conclusion: MLVA16 analysis of 54 Brucella isolates showed high level polymorphism in their genotypes. Only two genotypes, each observed in seven isolates, were related to one another and only one of these genotypes were found in to two separate regions.

Primer caso confirmado de brucelosis humana por Brucella melitensis, una zoonosis presente en Colombia / First confirmed case of human brucellosis by Brucella melitensis, a zoonosis present in Colombia

Resumen La brucelosis, principal zoonosis a nivel mundial tiene alta prevalencia en varios países de Latinoamérica. Se asocia con la exposición a ganado infectado por distintas especies del género Brucella. B. melitensis la más virulenta para el humano, causa con frecuencia complicaciones de predominio osteoarticular. En Colombia se cree que la infección por B. melitensis es una entidad ausente, a pesar de su plausibilidad biológica en nuestro contexto; sin embargo, son escasos los estudios sobre su ocurrencia y mínimo el índice de sospecha de la enfermedad, por lo cual creemos está subdiagnosticada. Presentamos el primer caso confirmado de brucelosis por B. melitensis en Colombia en una joven embarazada, con diagnóstico incidental, en quien el análisis retrospectivo de su cuadro clínico alertó sobre puntos clave que pueden impactar en el diagnóstico y tratamiento oportuno de la enfermedad. Se plantean preguntas de prevalencia real de esta entidad en Colombia.

Summary Brucellosis, the principal zoonoses globally is highly prevalent in different countries of Latin America. It is associated with the exposition of livestock infected with different Brucella species, being B. melitensis the most virulent for humans, and frequently causing osteoarticular complications. In Colombia it is believed that B. melitensis infection is an absent entity, despite its biological plausibility in our context; however, there are few studies on its occurrence and a minimum index of suspicion of the disease, which is why we believe it is underdiagnosed. We present the first confirmed case of brucellosis by B. melitensis in Colombia diagnosed in a young pregnant patient, with an incidental diagnosis, in whom a retrospective analysis of her clinical outcome warned of key points that may impact on the diagnosis and timely treatment of the disease. We present several questions surrounding the real prevalence of this entity in Colombia.

Interaction between Brucella melitensis 16M and small ubiquitin-related modifier 1 and E2 conjugating enzyme 9 in mouse RAW264.7 macrophages

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.

Caracterización de la variabilidad genética de cepas de campo de Brucella canis aisladas en Antioquia / Characterization of the genetic variability of field strains of Brucella canis isolated in Antioquia

Brucella canis, un patógeno intracelular facultativo, es responsable de la brucelosis canina, una enfermedad zoonótica que afecta a los caninos y al hombre. En los primeros causa abortos y fallas reproductivas; en el ser humano genera síntomas inespecíficos. En el año 2005 se demostró la presencia de B. canis en Antioquia (Colombia). Las cepas halladas se identificaron como tipo 2. La secuenciación del genoma completo de una cepa de campo denominada Brucella canis str. Oliveri mostró indels específicos de especie; a partir de estos se buscó conocer características genómicas de las cepas de B. canis aisladas y establecer relaciones filogenéticas, así como el tiempo de divergencia de la cepa Oliveri. Se realizó PCR convencional y secuenciación de 30 cepas de campo, se identificaron 5 indels reconocidos en B. canis str. Oliveri, se empleó ADN de Brucella suis, Brucella melitensis y cepas vacunales de Brucella abortus como controles. Se determinó que las cepas de campo estudiadas comparten 4 de los 5 indels de la cepa Oliveri, lo que indica la presencia de más de una cepa de B. canis circulando en la región. El análisis filogenético se realizó con 24 cepas de Brucella mediante secuencias concatenadas de genes marcadores de especie. Se probó la hipótesis del reloj molecular y adicionalmente se realizó test de tasas relativas de Tajima. De esta manera se demostró que la cepa Oliveri, al igual que las otras cepas de B. canis analizadas, divergen de B. suis. Se rechazó la hipótesis del reloj molecular entre las especies de Brucella y se demostró una tasa de evolución y una distancia genética similar entre las cepas de B. canis.

Brucella canis is a facultative intracellular pathogen responsible for canine brucellosis, a zoonotic disease that affects canines, causing abortions and reproductive failure; and the production of non-specific symptoms in humans. In 2005 the presence of B. canis in Antioquia was demonstrated and the strains were identified as type 2. The sequencing of the genome of a field strain denoted Brucella canis str. Oliveri, showed species-specific indel events, which led us to investigate the genomic characteristics of the B. canis strain isolated and to establish the phylogenetic relationships and the divergence time of B. canis str. Oliveri. Conventional PCR sequencing was performed in 30 field strains identifying 5 indel events recognized in B. canis str. Oliveri. ADN from Brucella suis, Brucella melitensis and vaccine strains from Brucella abortus were used as control, and it was determined that all of the studied field strains shared 4 out of the 5 indels of the sequenced Oliveri strain, indicating the presence of more than one strain circulating in the region. Phylogenetic analysis was performed with 24 strains of Brucella using concatenated sequences of genetic markers for species differentiation. The molecular clock hypothesis and Tajima's relative rate test were tested, showing that the Oliveri strain, similarly to other canis species, diverged from B. suis. The molecular clock hypothesis between Brucella species was rejected and an evolution rate and a similar genetic distance between the B. canis were demonstrated.

Serological study of brucellosis in Argentine Creole sheep / Estudio serológico de brucelosis en ovejas criollas de Argentina

Ovine cattle was introduced into America during the Spanish conquest with the second journey of Columbus to the Antilles and was disseminated throughout the region. In 1587, sheep were introduced into Argentina, later developing into the "Creole" breed. We selected 486 animals from different Argentine provinces with the aim of determining the serological status of brucellosis caused by Brucella melitensis and Brucella ovis. For the detection of antibodies against smooth Brucella spp., the Rose Bengal test (RBT) was performed as screening test while the serum agglutination test (SAT) and 2 mercapto-ethanol (2ME) were run as a confirmatory technique. Moreover, for the detection of antibodies against rough Brucella spp., we used the rapid slide agglutination test (RSAT) for screening and an indirect ELISA (IELISA) as confirmatory assay. This study showed that the total positive percentage of brucellosis due to B. ovis was 2.9%. Excluding the animals mixed with the Suffolk breed; seropositivity would be 0.6%. All animals tested negative for brucellosis caused by B. melitensis.

El ganado ovino fue introducido en América durante el segundo viaje de Colón a las Antillas y rápidamente se diseminó por la región. En 1587 entraron a la Argentina las primeras ovejas, estas se adaptaron a las condiciones naturales y se desarrolló así la denominada «oveja criolla¼. Se seleccionaron 486 animales de diferentes provincias para conocer el estado serológico de la enfermedad producida por Brucella melitensis y Brucella ovis. Para detectar anticuerpos anti-Brucella en fase lisa se utilizó la prueba de rosa de Bengala (RBT) como tamiz y como confirmatorias seroaglutinación lenta en tubo (SAT) y 2-mercaptoetanol (2ME). En fase rugosa se realizó como tamiz la prueba rápida de aglutinación (RSAT) y ELISA indirecto como prueba confirmatoria (IELISA). Este estudio mostró un porcentaje de seropositividad de brucelosis causada por B. ovis del 2,9%. Excluyendo los animales cruzados con la raza Suffolk, la seropositividad sería del 0,6%. Todos los animales fueron negativos a brucelosis causada por B. melitensis.

An Imported Case of Brucella melitensis Infection in South Korea

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.

An Imported Case of Brucella melitensis Infection in South Korea

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.

Membranoproliferative glomerulonephritis in a patient with chronic brucellosis

Brucellosis is the most common zoonotic disease in Greece, with an endemic distribution and can affect any organ. Infiltration of the renal parenchyma causes acute and chronic interstitial nephritis with granulomas, whereas renal glomeruli are rarely affected. The disease has been sporadically reported, and it causes various histopathologic patterns. Herein, we describe the case of a 39-year-old stock breeder with a history of recurrent episodes of bacteremia caused by Brucella melitensis over a period of 3 years. Two months after the last episode of bacteremia, he presented with mild renal insufficiency, nephrotic range proteinuria, and microscopic hematuria. A renal biopsy revealed membranoproliferative glomerulonephritis with a pattern of focal-segmental nodular sclerosis and moderate tubulointerstitial fibrosis. The patient received antimicrobial and corticosteroid therapy with partial remission of the nephrotic syndrome.

A Novel PCR Assay for Detecting Brucella abortus and Brucella melitensis

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.

Epidemiología de la brucelosis caprina y ovina en la provincia de Formosa, Argentina / Epidemiology of caprine and ovine brucellosis in Formosa province, Argentina

Se realizó un estudio epidemiológico de brucelosis en 516 majadas caprinas o mixtas (caprinos/ovinos) de las 3 regiones agroecológicas de la provincia de Formosa, Argentina. Mediante las pruebas de aglutinación en placa con antígeno tamponado y de fijación del complemento en suero se estudiaron un total de 25.401 caprinos y 2.453 ovinos. Además, se realizaron cultivos bacteriológicos y PCR en muestras de leche de cabras de 3 majadas con brucelosis y abortos recientes. Se detectó brucelosis en 4 de los 9 departamentos de la provincia, la prevalencia global fue del 2 % y la intrapredial varió entre el 1 y el 40%. La proporción de majadas positivas fue del 3,6, el 12 y el 36 % para las regiones este, centro y oeste, respectivamente. Se aisló Brucella melitensis bv. 1 de cabras por primera vez en la provincia. La PCR amplificó fragmentos esperados de 827 pb correspondiente al gen omp2ab (Brucella spp.) y de 731 pb correspondiente al inserto IS711 (B. melitensis). La detección de anticuerpos en ovinos que cohabitan con caprinos sugiere que las infecciones habrían sido causadas por B. melitensis, lo que constituye un riesgo adicional para la salud pública. Los programas de control y erradicación de la brucelosis deberían considerar las majadas mixtas como una sola unidad epidemiológica. Los resultados indican que la brucelosis por B. melitensis bv. 1 es altamente endémica en las regiones centro y oeste de la provincia de Formosa.

An epidemiological study of brucellosis was carried out in 516 goats and mixed flocks (goat/sheep) from the three agro-ecological regions of Formosa province, Argentina. Serum samples from a total of 25401 goats and 2453 sheeps were analyzed using buffered plate agglutination test (BPAT) and complement fixation test (CFT). Bacteriological and PCR analyses on milk samples from goats in three flocks with a history of brucellosis and recent abortions were also performed. Brucellosis was detected in four of the nine departments of the province with an overall prevalence of 2 % and an intra-flock prevalence ranging between 1 % and 40 %. The proportion of infected flocks was 3.6 %, 12 % and 36 % for the eastern, central and western regions, respectively. Brucella melitensis bv. 1 was isolated efrom goats for the first time in the province. The expected fragments of 827 bp from the omp2ab gene (Brucella spp.) and 731 bp from the insert IS711 (B. melitensis) were amplified by PCR. Detection of antibodies by BPAT and FCT in sheep cohabiting with goats suggests that infections could have been caused by B. melitensis, posing an additional risk to public health. Control and eradication programs for brucellosis should consider mixed flocks as a single epidemiological unit. The results indicate that brucellosis by B. melitensis bv1 is highly endemic in the central and western regions of Formosa province.

First Case of Human Brucellosis Caused by Brucella melitensis in Korea

No abstract available.

Brucella melitensis es un agente de la artritis pediatrica: Serie de casos / Brucella melitensis is an agent in pediatric arthritis: Case series

La brucelosis es un importante problema de salud y, en algunas regiones, es endémica. En ella, es muy frecuente observar un compromiso osteoarticular (20%-60%). En regiones donde es endémica, al analizar la etiología de la artritis, hay que pensar en esta entidad. En este artículo, se informan los casos de 12 pacientes; 11 de ellos eran varones, y la media de edad fue de 12 años (intervalo: de 2 a 17 años). Se determinó el compromiso de la cadera en 9 pacientes; de la articulación sacroilíaca, en 2; y de la rodilla, en 1. Todos los pacientes tenían artralgia; 8 pacientes tuvieron fiebre; y 1 paciente, leucocitosis. A un paciente se lo sometió a un tratamiento quirúrgico debido a artritis séptica. La infección puede derivar en complicaciones graves, especialmente en el compromiso del aparato locomotor. Debe tenerse en cuenta que, mediante el diagnóstico y tratamiento tempranos, es posible prevenir las complicaciones.

Brucellosis is a significant health problem and is endemic in some regions. Osteoarticular involvement is seen most frequently in brucellosis (20-60%). In regions where brucellosis is endemic, it should be considered in the etiology of arthritis. We report the 12 cases, were 11 males with a mean age of 12 years (range, 2-17 years). Involvement was determined in the hip in 9 cases, the sacroiliac joint in 2 and the knee in 1. All the cases had arthralgia, fever was determined in 8 cases and leukocytosis in one case. Surgical treatment was applied to one case because of septic arthritis. Serious complications of the infection may be encountered and particularly in musculoskeletal system involvement, it should be kept in mind that complications can be prevented by early diagnosis and treatment

[Simultaneous identification and comparison of pathogenic Brucella species in human serum and blood samples by means of multiplex PCR and confirmation of the results by PCR-RFLP]

There are few studies on the isolation of Brucella abortus and Brucella inelitensis from patients' sera. The aim of this study was simultaneous isolation of pathogenic brucella species from human serum samples by multiplex PCR method. 50 blood and serum samples isolated from patients with clinically suspected brucellosis were inoculated into brucella agar medium and we used rnultiplex-PCR, with three primers to detect brucella species. We incubated 0.5 ml of serum in Brucella broth for 72 hours at 37 °C in 5% carbon dioxide. To confirm the results of PCR, the PCR products were restricted by restriction enzymes, TaqI and RsaI. From 50 blood samples 4 [8%] cultures were positive. Using biochemical tests and after determination of the characteristics of the positive cases, they were identified as B. melitensis. After multiplex PCR, 9 cases [18%] were positive and 41 cases [82%] were negative. Among the positive sera, B. inelitensis was identified in 7 cases [78%] and B. abortus in 2 cases [22%]. Of so blood samples, 5 [10%] were positive and 45 [90%] were negative for B. melitensis. All the results were confirmed by PCR-RFLP. Our study showed that multiplex PCR method was simple, rapid and is more sensitive for isolation of brucella from serum [especially B. abortus and B. inelitensis] in comparison to complete blood

Identification of Brucella melitensis isolates originating from Mongolia and diagnostic real-time PCR evaluation using a specific SNP / 대한수의학회지

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.

Molecular detection of Brucella species in patients suspicious of Brucellosis from Zanjan, Iran

Brucella is an intracellular pathogen capable of infecting animals and humans. The aim of this study was to identify Brucella spp in sera of high risk individuals by a polymerase chain reaction (PCR)-based method. A total of 180 patients suspected to have Brucellosis were examined by serological tests. To establish a PCR protocol for diagnosis of active brucellosis, DNA was extracted from the serum samples by using a commercial kit. PCR amplification was done for detection of Brocella DNA using BCSP31 target gene and IS711 locus. The PCR assay showed that an amplicon of 223 bp was obtained in 73.8% (133/180) of the tested sera using primers (B4/B5) derived from a gene encoding the 31-kDa Brucella abortus antigen. In another PCR, an amplicon of 498 bp was obtained in 63.8% (115/180) of the samples using Brucella abortus-specific primers derived from a locus adjacent to the 3'-end of IS711, and also an amplicon of 731 bp was produced in 4.4% (8/180) of the tested samples using Brucella melitensis-specific primers. When the Wright method was used as a gold standard, the sensitivity and specificity of the PCR technique for genus identification were found to be 96 and 80.7%, respectively. However, the sensitivity value obtained with the species-specific PCR method was 82%, and specificity was similar to that previous reported. This is the first report of a high frequency of Brucella abortus in patients suspicious of Brucellosis from the Zanjan province.

Serum Levels of Interferon Gamma in Patients with Brucellosis in a Saudi Hospital.

Background: Brucellosis is a major zoonotic disease that is endemic in Saudi Arabia and it remains a major health problem that has not been eradicated in the country yet. Place and Duration of Study: This retrospective study was conducted in a Saudi Hospital at Al Madinah city during the period of 1 November, 2010 to 31 October, 2011. Methodology: All sera of patients suspected to have brucellosis (n= 65) and 18 healthy subjects were tested for brucella antibody using slide latex agglutination (SAT) and ELISA. Quantitation of IFN-ɣ was also done using ELISA. Results: Brucellosis was detected in all age groups but the incidence was higher and reached 33.3% in age group (40- <50) years with average of 43.9±2.53 years. Male to female ratio in infected patients was 2:1 by using SAT. The incidence of seropositive cases was high (80.1%) in the three months (April, May and June), with the highest peak in May (46.7%). Drinking raw milk was the most encountered risk factor with a prevalence of 66.1% followed by consumption of milk products (11.9%). The most prevalent species among the examined cases was B. melitensis (93.3%). Among the studied cases, 60 cases (92.3%) were serologically positive for brucellosis by SAT. Among the 60 cases yielding significant titers against brucella, 14 sera (23.3%) had agglutinin levels of 1:80, 34 sera (56.7%) had titers of 1:160 and 12 sera (20%) had titers of 1:320. By estimating IgM and IgG levels in the sera of examined cases using ELISA, 52 cases (80%) had brucellaIgM while 42 cases (64.6%) had brucella IgG. Sensitivities of SAT, IgM ELISA and IgG ELISA were 91.5%, 88.1% and 71.2%, respectively compared with combined ELISA. Mean IFN-ɣ levels ± SD in the subacute phase was 136.7±70.07pg/ml, 120.2±54.25pg/ml in the acute phase, and 121.3±51.09 pg/ml in the chronic phase of brucellosis. Conclusion: The sensitivity and specificity of ELISA to diagnose human brucellosis was higher when combined ELISA (IgM/IgG or both) was used. Mean IFN-ɣ levels were lower, but not significantly, in the chronic phase of the disease than in the sub acute phase and healthy subjects.

Study on the epidemiologic characteristic of Brucella melitensis isolated in Guizhou province in 2010-2012 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the genetic and epidemiologic characteristic of Brucella (B.) melitensis strains isolated in Guizhou province in 2010-2012.</p><p><b>METHODS</b>B. 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.</p><p><b>RESULTS</b>Six 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.</p><p><b>CONCLUSION</b>PCR 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.</p>

Brucella melitensis infection of ventriculo-peritoneal shunt: a form of neurobrucellosis manifested as gastrointestinal symptoms

A report of a 9 year-old child with Myelomeningocele who has a ventriculo-peritoneal [VP] shunt presented with gastrointestinal symptoms and peritonitis. The patient had no cNS symptoms but the cerebrospinal fluid was positive for Brucella melitensis. We discuss neurobrucellosis in children, its various presentations, complications and challenges in treatment, choice of antibiotics and duration

Collapsed L4 Vertebral Body Caused by Brucellosis

Brucellosis is caused by gram-negative, aerobic, non-motile, facultative, intracellular coccobacilli belonging to the genus Brucella. A 50-year-old man working as an employee was admitted to neurosurgery clinic with severe low back, radicular right leg pain and hypoesthesia in right L4-5 dermatomes for 2 months. Brucella tube agglutination (Wright) test was positive in serum sample of the patient with a titer of 1/640. Brucella melitensis was isolated from blood culture. X-ray and MRI of the lomber spine showed massive collapse of L4 vertebral body. Neural tissue was decompressed and then posterior L3-5 short segment transpedicular screw fixation and stabilization was performed. Brucella melitensis was isolated from microbiologic culture of pathologic specimen. Antibiotic therapy was given as doxycycline 200 mg/day and rifampicin 600 mg/day for 6 months. Brucellosis is a systemic zoonotic infection and still an important public health problem in many geographical parts of the world. Vertebral body collapse caused by brucellosis occurs very rarely but represents a neurosurgical emergency because of its potential for causing rapidly progressive spinal cord compression and permanent paralysis. Neurosurgeons, emergency department personnel as well as infectious disease specialists should always keep a high index of suspicion and include brucellosis in the differential diagnosis of vertebral body collapse.