Molecular detection of Brucella abortus in wild and captive felids

Abstract Purpose Brucellosis is a zoonotic disease of great public health importance. In wild animals, Brucella abortus is one of the most diagnosed species, mainly in enzootic environments where domestic animals share the same environment. B. abortus is common in environments shared by cattle, wild, and domestic animals. This study aimed to detect the presence of B. abortus DNA in free-ranging and captivity felids at Mato Grosso State, Brazil. Method Polymerase chain reaction, based on the genetic element IS711, was performed in blood samples collected from 23 free-ranging and captive felids. The species represented include Leopardus colocolo, Leopardus pardalis, Leopardus wiedii, Panthera onca, Puma concolor, and Puma yagouaroundi. Results DNA amplification of B. abortus was observed in only one captive P. concolor (4.34%). Conclusion The detection of this pathogen in captive animals using molecular tools demonstrates the importance of monitoring, as it raises concerns about the possibility of transmission between humans and wild and domestic animals, especially in regions of vast biodiversity, such as in the State of Mato Grosso, Brazil.

First study of Brucella ovis antibodies in purebred sheep flocks in the State of Parana, Brazil / [Primeiro estudo de anticorpos para Brucella ovis em rebanhos de ovinos puros de Origem, no estado do Paraná, Brasil]

Brucella ovis, a non-zoonotic species, is the etiological agent of ovine brucellosis, an infectious disease of clinical or subclinical occurrence in sheep flocks. Until then, there is no serological study of anti-Brucella ovis antibodies in purebred sheep herds. This study aimed to determine the presence of anti-Brucella ovis antibodies in purebred sheep flocks with breeding purposes from Parana State. Blood samples from 728 animals, of which 563 were females and 165 males, between 8 and 56 months of age from the six major sheep producing mesoregions of Parana, were submitted to detection of anti-Brucella ovis antibodies by the Agar Gel Immunodiffusion technique using an antigen from the bacteria Brucella ovis (Reo 198). The results indicate the presence of this disease in purebred sheep from Parana State in a low occurrence of 0.27% (2/728). The only two positive animals were rams, Santa Inês breed, from the same flock in the East Center region of Parana, without clinical disease. In conclusion, Brucella ovis is present in purebred sheep in Parana State, Brazil, and this low occurrence may have occurred due to rigorous breeding systems that may contribute to reduce the transmission of this disease.(AU)

Brucella ovis, espécie não zoonótica, é o agente etiológico da brucelose ovina, doença infecciosa de ocorrência clínica ou subclínica. Atualmente, não existe estudo sorológico de anticorpos anti-Brucella ovis em rebanhos de ovinos puros de origem. Este estudo teve como objetivo determinar a presença de anticorpos anti-Brucella ovis em rebanhos ovinos de raça pura de origem, com fins reprodutivos do estado do Paraná. Amostras de sangue de 728 animais, sendo 563 fêmeas e 165 machos, entre oito e 56 meses de idade, pertencentes a seis principais mesorregiões produtoras de ovinos no Paraná, foram submetidas à detecção de anticorpos anti-Brucella ovis pela técnica de imunodifusão em ágar gel usando-se um antígeno da bactéria Brucella ovis (Reo 198). Os resultados indicam a presença da doença em ovinos puros de origem do estado do Paraná em baixa ocorrência de 0,27% (2/728). Os dois únicos animais positivos foram reprodutores da raça Santa Inês, do mesmo rebanho da região Centro Leste do Paraná, sem manifestação clínica. Em conclusão, Brucella ovis está presente em ovinos puros de origem no estado do Paraná, e essa baixa ocorrência pode ter ocorrido devido a sistemas rigorosos de criação, que podem contribuir para a redução da transmissão dessa doença.(AU)

Estudio de casos confirmados de brucelosis humana en Puebla, México / Study of confirmed cases of human brucellosis in Puebla, Mexico

INTRODUCCIÓN: La brucelosis humana es un problema zoo-sanitario global. México es uno de los principales países que se ven afectados, el diagnóstico oportuno y las pruebas serológicas confirmatorias son la base para la detección. OBJETIVO: Conocer la frecuencia de casos confirmados de brucelosis en diferentes Unidades de Medicina Familiar del Instituto Mexicano del Seguro Social en el estado de Puebla, México. PACIENTES Y MÉTODOS: Estudio transversal en pacientes de ambos sexos, adultos y pediátricos, con manifestaciones clínicas sugestivas de brucelosis; se realizaron pruebas serológicas para el diagnóstico confirmatorio. RESULTADOS: De un total de 77 pacientes, se obtuvieron 39 (50,6%) casos positivos, 21 (27,3%) casos saliendo de la infección, 9 (11,7%) negativos y 8 (10,4%) con memoria inmunológica; de los casos positivos, 32 (82,1%) eran adultos y 30 (76,9%) fueron del género femenino. CONCLUSIÓN: Del total de muestras, la mitad tuvo diagnóstico de brucelosis, las manifestaciones clínicas de los pacientes estudiados fueron inespecíficas, lo cual resalta la importancia del diagnóstico de laboratorio.

BACKGROUND: Human brucellosis is a global health problem. Mexico is one of the main countries affected; timely diagnosis and serological tests are the basis for detection. AIM: To know the frequency of confirmed cases of brucellosis in different of Family Medicine Units of the Mexican Social Security Institute in the state of Puebla, Mexico. METHODS: Cross-sectional study in patients of both genders, adults and pediatrics, with clinical manifestations suggestive of brucellosis; serological tests were performed for the confirmatory diagnosis. RESULTS: Out of a total of 77 patients, 39 (50.6%) were positive, 21 (27.3%) cases coming out of infection, 9 (11.7%) were negative and 8 (10.4%) were defined with immunological memory; of positive cases, 32 (82.1%) were found in the adult group and 30 (76.9%) were female. CONCLUSION: Around half of samples were confirmed as brucellosis, the clinical manifestations of the patients studied were non-specific, which highlights the importance of laboratory diagnosis.

Prevalência de brucelose bovina na região noroeste do estado do Paraná de 2014 á 2016 / Bovine brucellosis prevalence in the northwest region of the state of Paraná from 2014 to 2016 / Prevalencia de brucelosis bovina en la región noroeste del estado de Paraná de 2014 a 2016

Dentre as zoonoses responsáveis pelas maiores perdas socioeconômicas na bovinocultura brasileira, a brucelose possui significativo destaque, sendo considerada uma antropozoonose causada pelas bactérias do gênero Brucella spp. Essas bactérias possuem potencial para infectar diversos mamíferos, inclusive, existem registros de brucelose em cetáceos (baleias e golfinhos). Nos bovinos, as principais maneiras de transmissão se dão pelas descargas bacterianas presentes nos restos de materiais provenientes de abortos de vacas positivas para brucelose, mas também pode ser transmitida pelo leite, cópula, fômites, solo e até a água. As vacas continuam liberando as bactérias por cerca de 30 dias após o parto ou aborto. Para diagnóstico são utilizados testes diretos ou indiretos preconizados pelo Programa Nacional de Controle e Erradicação da Brucelose e da Tuberculose Animal (PNCEBT). O objetivo deste trabalho foi avaliar o perfil epidemiológico de Brucelose bovina na região noroeste do estado do Paraná de 2014 a 2016. Entre os anos de 2014 a 2016, foram utilizados dados de exames realizados por Médico Veterinário devidamente registrado e habilitado para a realização do diagnóstico. Entre bovinos machos e fêmeas foram testados 12.640 animais, na região noroeste do estado Paraná, sendo o teste utilizado o do antígeno acidificado tamponado (AAT). 26 animais foram positivos para brucelose, perfazendo um percentual de 0,2 % do rebanho analisado. Apesar dos baixos índices de animais infectados pela Brucella spp. apresentados nessa pesquisa o controle e uma possível erradicação da brucelose se tornam indispensáveis devido à fácil transmissão e às perdas econômicas provocadas, principalmente pelos abortos ocorridos nos últimos trimestres das gestações.(AU)

Brucellosis features among the zoonoses responsible for greatest socioeconomic losses in the Brazilian cattle farming. Considered an anthropozoonosis, its etiological agents are bacteria of the Brucella spp genus. Such bacteria have the potential to infect several mammals, with records of brucellosis even in cetaceans (whales and dolphins). In cattle, it is mainly transmitted through bacterial discharges present in the remains of materials coming from abortions tested positive for brucellosis. Nevertheless, it can also be transmitted by milk, copulation, blood transfusions, fomites, soil and even water. Cows continue releasing the bacteria for about 30 days after calving or abortion. Direct or indirect tests as recommended by the National Program for the Control and Eradication of Brucellosis and Animal Tuberculosis (PNCEBT) can be used for diagnosis. The objective of this study was to assess the epidemiological profile of bovine brucellosis in the northwestern region of the State of Paraná from 2014 to 2016. Between 2014 and 2016, data from examinations performed by a veterinarian duly registered and qualified to perform the diagnosis were used. A total of 12,640 animals in the northwest region of the state of Paraná were tested, both male and female, using the acidified buffered antigen (ABA). A total of 26 animals tested positive for brucellosis, which corresponds to 0.2% of the studied herd. Despite the low rates of animals infected with Brucella spp. presented in this study, the control and possible eradication of brucellosis is essential due to its easy transmission and extensive economic losses mainly caused by abortions occurring during the last quarters of pregnancies.(AU)

Entre las zoonosis responsables por las mayores pérdidas socioeconómicas en la bovinocultura brasileña, la brucelosis posee significativo destaque, es considerada una antropozoonosis causada por bacterias del género Brucella spp. Esas bacterias poseen potencial para infectar diversos mamíferos, incluso, existen registros de brucelosis en cetáceos (ballenas y delfines). En los bovinos, las principales formas de transmisión se dan por las descargas bacterianas presentes en los restos de materiales provenientes de abortos de vacas positivas para brucelosis, también puede ser transmitida por la leche, cópula, fómites, suelo e incluso el agua. Las vacas continúan liberando las bacterias durante unos 30 días después del parto o aborto. Para el diagnóstico se utilizan pruebas directas o indirectas preconizadas por el Programa Nacional de Control y Erradicación de Brucelosis y de la Tuberculosis Animal (PNCEBT). El objetivo de esta investigación ha sido evaluar el perfil epidemiológico de Brucelosis bovina en la región noroeste del estado de Paraná de 2014 a 2016. Entre los años de 2014 a 2016 se utilizaron datos de exámenes realizados por Médico Veterinario debidamente registrado y habilitado para la realización del diagnóstico. Entre bovinos machos y hembras se probaron 12.640 animales, en la región noroeste del estado de Paraná, la prueba utilizada fue antígeno acidificado tamponado (AAT). 26 animales atestaron positivos para brucelosis, lo que significó el 0,2% del rebaño analizado. A pesar de los bajos índices de animales infectados por Brucella spp. presentados en esa investigación, el control y una posible erradicación de la brucelosis se vuelven indispensables debido a su fácil transmisión y las pérdidas económicas provocadas, principalmente por los abortos ocurridos en los últimos trimestres de las gestaciones.(AU)

Establishment and application of a rapid PCR detection method of Brucella spp / 国际检验医学杂志

Objective To establish a polymerase chain reaction (PCR) method for detecting Brucella spp .and to evaluate its ap‐plication .Methods The PCR primer aiming at the outer membrane protein(omp‐2) coding gene were designed and the PCR method for detecting Brucella spp .was established .By using DNA of 19 kinds of common bacteria and Brucella spp .,the specificity and sensitivity were evaluated .Three strains of suspected Brucella spp .were performed the nucleic acid detection .Then ,3 suspected strains of Brucella spp .were amplified by the PCR method with omp‐2 gene ,and the results were compare to the those by the cul‐ture method .Results The established Brucella spp .PCR detection method could only detect the Brucella spp .positive strain ,the control bacterium DNA did not appear the target band ;the sensitivity was 100 copies/response;the PCR method detected omp‐2 gene band in 3 strains of suspected Brucella spp .,which was identified as Brucella spp .and was same to the results by the isolation culture method .Conclusion The established PCR method for detecting Brucella spp .is accurate and rapid compare with the isola‐tion and culture method ,which is suitable for the rapid detection of brucellosis epidemic situation .

Seroprevalence for brucellosis and leptospirosis in dogs from Belém and Castanhal, State of Pará, Brazil / Soroprevalência para brucelose e leptospirose em cães de Belém e Castanhal, Pará, Brasil

Brucellosis and leptospirosis are widely spread bacterial infections and dogs are the most important source of infection and reservoir for diseases. Dogs can disseminate the agents in the environment and transmit them to humans and/or other animals. The objective of this study was assess the occurrence of reactive to antibodies anti-Leptospira spp., Brucella canis and B. abortus in Belém and Castanhal, State of Pará, Amazon, Brazil. A total of 156 samples were randomly collected in the city of Belém and 158 samples in Castanhal. The anti-B. canis antibodies research was performed by Agar Gel Immunodiffusion (AGID) with and without 2-mercaptoethanol serum treatment (AGID-2ME). To assess the anti-B. abortus antibodies, the technique of Fast Seroagglutination with buffered acidified plate antigen (BAPAT) was used. For anti-Leptospira spp. antibodies research, the Microscopic Agglutination Technique (MAT) was used. No animal reacted to Brucella abortus and one animal was reactive to B. canis at the AGID, but it was negative to the AGID-2ME test. Seventeen percent of dogs (47/274) presented anti-Leptospira spp. antibodies, with prevalence of serovar Canicola. The dogs from Belém and Castanhal are not source of infection for B. abortus and B. canis, however, they are reservoirs for different serovars of Leptospira spp.

Brucelose e leptospirose são enfermidades bacterianas amplamente disseminadas e o cão é importante fonte de infecção e reservatório para ambas, podendo eliminar o agente no meio ambiente, e transmiti-lo para humanos e/ou outros animais. O presente estudo teve como objetivo pesquisar a ocorrência de cães reagentes para anticorpos contra Leptospira spp., Brucella canis e B. abortus em Belém e Castanhal, Pará, Amazônia, Brasil. Foram colhidas de forma aleatória 156 amostras no município de Belém e 158 amostras em Castanhal. A pesquisa de anticorpos anti-B. canis foi realizada pela técnica de Imunodifusão em Gel de Ágar (IDGA) com e sem tratamento do soro com 2-mercaptoetanol (IDGA-2ME) e para pesquisa de anticorpos anti-B. abortus foi utilizada a técnica de Soroaglutinação Rápida com antígeno acidificado tamponado (AAT). Para pesquisa de anticorpos contra-Leptospira spp. utilizou-se a Técnica de Aglutinação Microscópica (MAT). Nenhum animal reagiu para Brucella abortus e um animal foi reagente para B. canis na IDGA, porém foi negativo na IDGA-2ME. Dezessete por cento dos cães (47/274) apresentaram anticorpos contra-Leptospira spp., com predominância do sorovar Canicola. Os cães de Belém e Castanhal não são fontes de infecção para B. abortus e B. canis, no entanto são reservatórios de diferentes sorovares de Leptospira spp.

Detección de Brucella spp. por un sistema inmunocromatográfico comercial, en muestras ambientales cubanas / Detection of Brucella spp using a commercial system in enviroment cuban samples

Introducción: actualmente en Cuba se desconoce la circulación de las especies de Brucella en el medio ambiente, por la inexistencia de métodos de laboratorio que permitan su identificación. Objetivos: detectar Brucella spp. en muestras ambientales cubanas aplicando un sistema inmunocromatográfico comercial. Métodos: se estudiaron 59 muestras ambientales de una zona endémica de brucelosis bovina y 50 muestras ambientales de zonas controladas de la enfermedad, durante el período diciembre de 2011 y hasta febrero de 2012. Se utilizó el sistema inmunocromatográfico directo de flujo lateral para Brucella spp. comercializado en Cuba. Resultados: el 52,5 por ciento (31/59) de las muestras ambientales de la zona endémica resultaron positivas para Brucella spp. Las muestras ambientales que presentaron el mayor porcentaje de positividad fueron el estiércol (62,5 por ciento ) y las del suelo cementado (26,9 por ciento ). Predominaron las reacciones fuertemente positivas en un 74,1 por ciento (23/31). Conclusiones: el sistema inmunocromatográfico comercial detecta un elevado porcentaje de Brucella spp. en muestras ambientales de la zona endémica cubana, lo que pudiera avalar su implementación en la red de laboratorios de Salud Pública y del Instituto de Medicina Veterinaria de Cuba. Los resultados de esta investigación deben complementarse con los aislamientos de brucelas en el medio ambiente, pacientes humanos y en animales(AU)

Introduction: Information about the circulation of Brucella species in the Cuban environment is not currently available due to the lack of laboratory methods for their identification. Objectives: Detect Brucella spp. in Cuban environmental samples using a commercial immunochromatographic system. Methods: A study was conducted of 59 environmental samples from an endemic zone for bovine brucellosis and 50 environmental samples from areas where the disease has been controlled, from December 2011 to February 2012. Use was made of the direct lateral flow immunochromatographic system for Brucella spp. commercially available in Cuba. Results: Of the environmental samples from the endemic zone 52.5 percent (31/59) tested positive for Brucella spp. The environmental samples showing the highest positivity were those taken from dung (62.5 percent) and cemented soil (26.9 percent). Strongly positive reactions predominated in 74.1 percent (23/31) of the samples. Conclusions: The commercial immunochromatographic system used detected a high percentage of Brucella spp. in environmental samples from the endemic zone, which could justify its implementation in the Public Health laboratory network and the Institute of Veterinary Medicine of Cuba. The results obtained should be complemented with Brucella isolates from the environment, human patients and animals(AU)

Varón marroquí con espondilodiscitis de larga evolución, orquitis y endocarditis / Moroccan man with longstanding spondylodiscitis, orchitis and endocarditis

La afectación osteoarticular es la complicación más frecuente de la brucelosis. La espondilodiscitis es una complicación grave, ya que suele estar asociada con la formación de abscesos. Se presenta el caso de un varón de 43 años de edad, natural de Marruecos, con síntomas, signos y hallazgos imagenológicos de espondilodiscitis de larga evolución, orquitis y endocarditis. Mediante la realización de pruebas de laboratorio complementarias se llega al diagnóstico de infección por Brucella spp. El presente caso busca concientizar a la comunidad médica sobre la importancia de considerar la infección por Brucella spp. como diagnóstico diferencial en pacientes con sospecha de espondilodiscitis infecciosa, especialmente en aquellos pacientes con antecedentes epidemiológicos, permitiendo así realizar un acertado y oportuno diagnóstico. Este caso resulta interesante porque nunca antes se había descrito la presencia simultánea de estas tres manifestaciones de la brucelosis en un paciente inmuno competente. (Acta Med Colomb 2015; 40: 166-168).

Osteoarticular involvement is the most common complication of brucellosis. Spondylodiscitis is a serious complication as it is often associated with abscess formation. The case of a 43 years old man born in Morocco with symptoms, signs and imaging findings of longstanding spondylodiscitis, orchitis and endocarditis is presented. Through the implementation of complementary laboratory tests, diagnosis infection by Brucella spp. was made.This case aims to raise awareness in the medical community about the importance of considering Brucella spp. as a differential diagnosis in patients with suspected infectious spondylodiscitis, especially in those patients with epidemiological background, allowing make a successful and timely diagnosis. This case is interesting because never before had been described the simultaneous presence of these three manifestations of brucellosis in an immunocompetent patient. (Acta Med Colomb 2015; 40: 166-168).

Soroprevalência de pseudorraiva, peste suína clássica e brucelose em suínos do estado do Piauí / Seroprevalence of pseudorabies, classical swine fever and brucellosis in swine in the state of Piauí

Este trabalho teve como objetivo determinar a soroprevalência de pseudorraiva, peste suína clássica (PSC) e brucelose suína em suínos do estado do Piauí, Brasil. Foram coletadas amostras sanguíneas de 384 suínos de criações intensivas e extensivas do estado. Anticorpos anti-Brucella spp. foram detectados pelo teste do antígeno acidificado tamponado e confirmados pelo teste 2-mercaptoetanol, enquanto a detecção de anticorpos contra os vírus da PSC e pseudorraiva foi realizada por ensaio imunoenzimático (ELISA), utilizando-se kits comerciais específicos. Anticorpos anti-Brucella spp. foram detectados em 1,04% (2/192) dos suínos de criações intensivas. Dos rebanhos avaliados, 0,78% (3/384) dos animais exibiram anticorpos contra o vírus da PSC, sendo 1,04% (2/192) de criações intensivas e 0,52% (1/192) de criações extensivas. Anticorpos contra o vírus da pseudorraiva foram detectados apenas em suínos de criação extensiva, com prevalência de 5,2% (10/192). Esses são os primeiros dados sobre a soroprevalência da brucelose suína, pseudorraiva e PSC em rebanhos do Piauí. A detecção de 10 amostras positivas para pseudorraiva causa preocupação sobre a possibilidade da circulação viral na população suídea desse estado e revela uma necessidade premente de realização de estudos mais extensos para melhor compreender a importância dessas enfermidades de notificação obrigatória em estados da região Nordeste brasileira.

This study aimed to determine the prevalence of Pseudorabies, Classical Swine Fever (CSF) and Swine Brucellosis in swine in the state of Piauí, Brazil. Blood samples were collected from 384 pigs from intensive and small outdoor systems in the state. Anti-Brucella spp. antibodies were detected by Buffered Acidified Antigen Test and positive results confirmed by 2-Mercaptoethanol Test. Detection of antibodies against CSF and Pseudorabies virus were performed by Enzyme-Linked Immunosorbent Assay (ELISA) using specific commercial kits. Only two samples (1.04% - 2/192) from the intensive system were seropositive to Brucella spp. In the evaluated herds, 0.78% (3/384) of animals had antibodies against CSF virus, two from outdoor pigs (1.04% - 2/192) and one from intensive systems (0.52% - 1/192). Antibodies against the Pseudorabies virus were detected only in outdoor pigs, with seroprevalence of 5.2% (10/192). This is the first report on seroprevalence of Pseudorabies, CSF and Brucellosis in hog farms in Piauí, Brazil. The detection of 10 positive cases raises a concern regarding Pseudorabies virus circulation in the swine population in the state and reveals a need for further studies to better understand the real situation and status of obligatory notified diseases in the swine herds in the Northestern states of Brazil.

Diseño y evaluación de una prueba de PCR para detección de Brucella spp. y Brucella abortus / Design and evaluation of a PCR test for detection of Brucella spp. and Brucella abortus / Desenho e avaliação de um teste de PCR para a detecção de Brucella spp. e Brucella abortus

Resumen Problema: el diagnóstico oportuno de la brucelosis, zoonosis que afecta ganado y trabajadores pecuarios, es difícil. Las pruebas serológicas Rosa de Bengala y ELISA, el hemocultivo y mielocultivo, tienen sensibilidades entre 15-70% dependiendo del estadío de la infección. El desarrollo de métodos diagnósticos rápidos, sensibles y específicos utilizando técnicas moleculares como la PCR, permite diagnosticar y tratar oportunamente esta enfermedad. Objetivo: Desarrollar y evaluar dos pruebas de PCR para detección de Brucella spp. y B. abortus. Materiales y métodos: se diseñaron y evaluaron iniciadores para detectar el gen ugpA utilizando Primer3, y otros reportados en la literatura. Se calculó sensibilidad y especificidad, usando 3 grupos (G) de muestras humanas y bovinas utilizando el teorema de Bayes: G1:30 muestras de suero y 30 de sangre de humanos sanos y 30 de suero y 30 de sangre de bovinos sanos, inoculadas con Brucella abortus; G2: 30 muestras de suero y 30 de sangre de humanos sanos y 30 de suero y de 30 sangre de bovinos sanos inoculadas con Salmonella Typhi, Escherichia coli, Klebsiella sp., Psudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumoniae y Enterococcus sp. G3: 60 muestras de suero y 60 sangre de bovinos y 60 muestras de suero y 60 sangre de humanos asintomáticos. Resultados: se obtuvo una sensibilidad y especificidad de detección de Brucella sp. y Brucella abortus del 100% en muestras humanas y bovinas. Conclusiones: por la alta sensibilidad y especificidad encontradas en las pruebas de PCR estudiadas, se recomienda continuar un estudio clínico de aplicación para evaluar su comportamiento en muestras de pacientes y animales infectados.

Abstract Problem: brucelosis is a zoonosis that affects livestock and livestock workers. This disease is characterized by the difficulties for its early diagnosis. Sensitivity of marrow and blood cultures as well as serological tests (Rose Bengal and ELISA) ranges between 15 and 70%, depending on the stage of infection. Development of rapid, sensitive, and specific diagnostic methods using molecular techniques such as PCR would allow timely diagnose and treatment of the disease. Objective: To develop and evaluate two PCR tests for detection of Brucella spp. and B. abortus. Materials and methods: primers to detect ugpA gene were designed and evaluated using Primer3 and others reported in the literature. Sensitivity and specificity were calculated for 3 groups (G) of human and bovine samples using Bayes' theorem. G1 consisted of healthy human and healthy bovine samples (30 serum and 30 blood samples of each species); bovine blood samples were inoculated with Brucella abortus. G2 consisted of healthy human and healthy bovine samples (30 serum and 30 blood samples of each); bovine blood samples were inoculated with Salmonella typhi, Escherichia coli, Klebsiella sp., Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumonia, and Enterococcus sp. G3 consisted of 60 serum and 60 blood samples from asymptomatic bovines, as well as 60 serum and 60 blood samples from asymptomatic humans. Results: sensitivity and specificity of the PCR test to detect Brucella sp. and Brucella abortus reached 100% in human and bovine samples. Conclusiones: considering the high sensitivity and specificity observed in the PCR tests studied, we recommend doing a follow up of the test in a clinical trial to evaluate its performance in infected humans and animals.

Resumo Problema: é difícil o diagnostico oportuno da brucelose, zoonose que afeta o gado e os trabalhadores pecuários. Os testes serológicos Rosa de Bengala e ELISA, e a hemocultura e mielocultura têm sensibilidades entre 15 e 70% dependendo do estádio da infecção. O desenvolvimento de métodos diagnósticos rápidos, sensíveis e específicos utilizando técnicas moleculares como a PCR, permite diagnosticar e tratar oportunamente esta doença. Objetivo: desenvolver e avaliar dois testes de PCR para a detecção de Brucella spp. e B. abortus. Materiais e métodos. Desenharam-se e avaliaram-se iniciadores utilizando o software Primer3 para detectar o gene ugpA, também se avaliaram outros iniciadores reportados previamente na literatura. Calculou-se a sensibilidade e especificidade do teste por PCR usando três grupos (G) de amostras humanas e bovinas utilizando o teorema de Bayes: G1: 30 amostras de soro e 30 de sangue de humanos sãos e 30 de soro e 30 de sangue de bovinos sãos, inoculadas com Brucella abortus; G2: 30 amostras de soro e 30 de sangue de humanos sãos e 30 de soro e de 30 sangue de bovinos sãos inoculadas com Salmonella Typhi, Escherichia coli, Klebsiella sp., Psudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumoniae e Enterococcus sp. G3: 60 amostras de soro e 60 de sangue de bovinos e 60 amostras de soro e 60 sangue de humanos assintomáticos. Resultados. Obteve-se uma sensibilidade e especificidade de detecção da Brucella spp. e Brucella abortus de 100% em amostras humanas e bovinas. Conclusões: pela alta sensibilidade e especificidade encontrada com o teste de PCR estudado, recomenda-se continuar um estudo clínico de aplicação para avaliar seu comportamento em amostras de pacientes e animais infetados.

Chemoprophylaxis and Serological Follow-Up in Brucella abortus Exposed Laboratory Workers / 감염과화학요법

The aerozolization is one of possible Brucella transmission mechanisms, particularly in air-borne exposed laboratory workers. In this study, seven laboratory workers were potentially exposed to B. abortus via aerosols. Two laboratory workers who sniffed an agar plate several times were considered to be at high risk for acquiring the disease, 5 workers who did not work directly with the strain were be considered at low risk of infection. Prophylactic antibiotics of doxycycline 200 mg/day and rifampin 600 mg/day were offered for 6 weeks in high risk workers and 3 weeks for low risk workers, respectively. Enrolled workers were advised to check for serological testing of Brucella species every 3 weeks during a total period of 12 weeks. Compliance with taking medicine was 57.1% (4/7) and compliance for completing the serological tests was 85.7% (6/7). None of the laboratory workers developed clinical disease or tested positive serologically during 3 months of serological testing and 1 year of clinical follow-up.

Chemoprophylaxis and Serological Follow-Up in Brucella abortus Exposed Laboratory Workers / 감염과화학요법

The aerozolization is one of possible Brucella transmission mechanisms, particularly in air-borne exposed laboratory workers. In this study, seven laboratory workers were potentially exposed to B. abortus via aerosols. Two laboratory workers who sniffed an agar plate several times were considered to be at high risk for acquiring the disease, 5 workers who did not work directly with the strain were be considered at low risk of infection. Prophylactic antibiotics of doxycycline 200 mg/day and rifampin 600 mg/day were offered for 6 weeks in high risk workers and 3 weeks for low risk workers, respectively. Enrolled workers were advised to check for serological testing of Brucella species every 3 weeks during a total period of 12 weeks. Compliance with taking medicine was 57.1% (4/7) and compliance for completing the serological tests was 85.7% (6/7). None of the laboratory workers developed clinical disease or tested positive serologically during 3 months of serological testing and 1 year of clinical follow-up.