Detection of Brucella ovis in ovine from Paraíba State, in the Northeast region of Brazil

To determine the presence of Brucella ovis in ovine from Paraíba State, in the Northeast region of Brazil, 80 animals slaughtered in the public slaughterhouse of Patos city were used. Before slaughter, blood samples were collected by jugular venopuncture from each animal, and after slaughter, testicles, epidydimus and uterus were aseptically collected. For the serological diagnosis of B. ovis and B. abortus infections, the agar gel immunodiffusion (AGID) and Rose Bengal (RBT) tests were carried out, respectively. In addition, microbiological culture and polymerase chain reaction (PCR) were performed on testicle, epidydimus and uterus samples. Six animals (7.5%) tested positive for the presence of B. ovis antibodies and all animals tested negative for the presence of B. abortus antibodies. One AGID-positive animal tested positive at uterine swab culture. PCR was able to amplify DNA of Brucella spp. from the pool of testicle, epidydimus and uterus samples from AGID-positive animals. This is the first report of isolation and detection of B. ovis DNA by PCR in ovine from the Northeast region of Brazil.

Evidência molecular de Brucella sp. em Ozotoceros bezoarticus (veado campeiro) do Pantanal Sul-Mato-Grossense

A presença de Brucella spp. entre animais silvestres pode influenciar a taxa de reprodução destes hospedeiros, além de atuarem como fonte de infecção natural para os animais domésticos e humanos. O objetivo deste estudo foi identificar a presença de Brucella spp. em 44 amostras de sangue de veado campeiro (Ozotoceros bezoarticus) do Pantanal do Sul-Mato-Grossense, utilizando a técnica de PCR. Observou-se que 20,4% (9/44) das amostras foram positivas. A sequência consenso de nucleotídeo obtida no sequenciamento do isolado de veado campeiro apresentou 514 pb e 95% de identidade com virB5 de B. abortus (best hits acesso nr AF226278, e-value 0.0), já na análise filogenética a amostra de Brucella isolada de veado campeiro apresentou-se muito próximo de B. suis. A alta porcentagem de amostras positivas sugere que a brucelose pode ser um problema entre os veados campeiros na área estudada e que estes animais podem representar riscos para outros animais domésticos e silvestres.

Brucella melitensis 16M: characterisation of the galE gene and mouse immunisation studies with a galE deficient mutant

The galE gene of Streptomyces lividans was used to probe a cosmid library harbouring Brucella melitensis 16M DNA and the nucleotide sequence of a 2.5 kb ClaI fragment which hybridised was determined. An open reading frame encoding a predicted polypeptide with significant homology to UDP-galactose-4-epimerases of Brucella arbortus strain 2308 and other bacterial species was identified. DNA sequences flanking the B. melitensis galE gene shared no identity with other gal genes and, as for B. abortus, were located adjacent to a mazG homologue. A plasmid which encoded the B. melitensis galE open reading frame complemented a galE mutation in Salmonella typhimurium LB5010, as shown by the restoration of smooth lipopolysaccharide (LPS) biosynthesis, sensitivity to phage P22 infection and restoration of UDP-galactose-4-epimerase activity. The galE gene on the B. melitensis 16M chromosome was disrupted by insertional inactivation and these mutants lacked UDP-galactose-4-epimerase activity but no discernible differences in LPS structure between parent and the mutants were observed. One B. melitensis 16M galE mutant, Bm92, was assessed for virulence in CD-1 and BALB/c mice and displayed similar kinetics of invasion and persistence in tissues compared with the parent bacterial strain. CD-1 mice immunised with B. melitensis 16M galE were protected against B. melitensis 16M challenge. Crown Copyright (C) 1999 Published by Elsevier Science B.V.

Risk factors and presence of antibodies to Brucella canis and smooth Brucella in dogs from the municipality of Araguaina, Tocantins, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Perfil de Aglutininas Anti-leptospira e anti-brucella e condições sanitárias de ovinos da região noroeste do Estado de São Paulo, Brasil

The sheep industry has become increasingly prominent in agribusiness, transforming the stage production of Brazil, and thus contributes more to the socio-economic development of the country. The work aimed to verify the occurrence of brucellosis and leptospirosis in sheep from northwestern São Paulo state. In addition to determining the prevalence of major Leptospira in the region and to trace the diagnosis of sheep breeding in this part of the country. All the 1222 sheep serum samples from 49 properties did not react serologically to evidentiary testing for brucellosis, compared to antigens of B. abortus and B. ovis used in the testing of 2-ME and IDGA, respectively. The Microscopic Agglutination Test (MAT) test revealed that 19.14% (232/1212) of samples were positive for one or more serovars, with titles ranging from 100 to 800. The most frequent serovar was hebdomadis in the region, with Sentot and Sherman (18.10%, 11.64% and 8.62%, respectively). By profiling the system of sheep farming in the region, we found that most herds are composed of more than one race being the main purpose is for the court. According to the scheme adopted immunoprophylactics there is a homogeneous set schedule. It adopts the use of anti-helminth, non-prescription veterinary antibiotic. There is also the presence of diarrhea and abortions and the lack of criteria for los ovinos on management, making it the need for emergency development of programs for disease control, schema immunoprophylactics adequate sanitation and hygienic measures in sheep breeding.

Assessment of genetic diversity of zoonotic Brucella spp. recovered from livestock in Egypt using multiple locus VNTR analysis

Brucellosis is endemic in most parts of Egypt, where it is caused mainly by Brucella melitensis biovar 3, and affects cattle and small ruminants in spite of ongoing efforts devoted to its control. Knowledge of the predominant Brucella species/strains circulating in a region is a prerequisite of a brucellosis control strategy. For this reason a study aiming at the evaluation of the phenotypic and genetic heterogeneity of a panel of 17 Brucella spp. isolates recovered from domestic ruminants (cattle, buffalo, sheep, and goat) from four governorates during a period of five years (2002-2007) was carried out using microbiological tests and molecular biology techniques (PCR, MLVA-15, and sequencing). Thirteen strains were identified as B. melitensis biovar 3 while all phenotypic and genetic techniques classified the remaining isolates as B. abortus (n = 2) and B. suis biovar 1 (n = 2). MLVA-15 yielded a high discriminatory power (h = 0.801), indicating a high genetic diversity among the B. melitensis strains circulating among domestic ruminants in Egypt. This is the first report of the isolation of B. suis from cattle in Egypt which, coupled with the finding of B. abortus, suggests a potential role of livestock as reservoirs of several zoonotic Brucella species in the region.

Pesquisa de aglutininas anti Brucella canis em soros humanos na cidade de São Paulo, Brasil

De 330 soros humanos examinados pela prova de soroaglutinação lenta em tubos, 4(1,21%) apresentaram aglutininas anti Brucella canis em diluição 1:100 (1 reagente com título 100, 2 reagentes com título 200 e 1 reagente com título 400).

Brucella canis: inquéritos sorológico e bacteriológico em população felina

De 134 soros de felinos domésticos examinados pela prova de soroaglutinação lenta em tubos, 4 (3%) foram positivos para Brucella canis, todos com título igual a 100. Não se obteve êxito na tentativa de isolamento de Brucella canis através de hemocultura desses animais.

Estudio de prospección serológica de brucelosis y leptospirosis canina

Las enfermedades infecciosas bacterianas del perro de mayor importancia en salud pública son la Brucelosis y la Leptospirosis canina. Ambas enfermedades se encuentran mundialmente difundidas. En nuestro país no se conocen datos concretos de prevalencia. Si bien las pérdidas que generan en criaderos comerciales son cuantiosas, no se comparan con el riesgo zoonótico que estas patologías representan para la comunidad. El agente etiológico de la Brucelosis en el perro, es principalmente B. canis , aunque también se describen infecciones con cepas de B. abortus, melitensis y suis.El diagnóstico de esta enfermedad es diferente según el tipo de cepa actuante. (...) El agente etiológico de la Leptospirosis en el perro es principalmente Leptospira canicola, pero también se afecta con otros serovares del género L. interrogans. Este animal juega un rol muy importante como diseminador del microorganismo en el medio ambiente y de allí la posibilidad de contagio al hombre, ya que la enfermedad se transmite por contacto directo con material contaminado como agua, tierra, vegetales contaminados con orina de animales infectados. (...) Objetivo general: Se plantea como objetivo del presente proyecto, determinar la tasa de infección a B. spp, y Leptospira interrogans en los perros ingresados al hospital de Clínica Animal de la Facultad de Agronomía y Veterinaria de la Universidad Nacional de Río Cuarto. Objetivos específicos: 1. Elaborar el antígeno para realizar la técnica de Inmunodifusión utilizada en el diagnóstico de Brucella canis. 2. Realizar la técnica de ID y compararla con los resultados obtenidos mediante técnicas de aglutinación capaces de detectar la infección por B. abortus. (BPA, Rosa de Bengala, 2ME, Lenta en tubo Wright). 3. Hemocultivo de animales serológicamente positivos a Brucella sp. 4. Determinar mediante la técnica de microaglutinación, las tasa de infección frente a distintos serovares de Letospira interrogans.

Ecoepidemiología de Chlamydophila psittaci, pecorum, pneumoniae, abortus en Córdoba. Eco-epidemiology of Chlamydophila psittaci, pecorum, pneumoniae, abortus in Córdoba.

Las Clamidias son bacterias patógenas de los animales de producción, de vida silvestre y de compañía. Además de las pérdidas económicas que producen las infecciones en los planteles de producción bovina, ovina, caprina, porcina y aves de corral, la mayoría de las especies tienen importancia zoonótica, pudiendo dar origen a infecciones graves, potencialmente letales en el ser humano. El orden Chlamydiales está integrado por bacterias que actúan como parásitos intracelulares obligados que desarrollan su ciclo de vida únicamente dentro de inclusiones citoplasmáticas. En este orden se encuentra la familia Chlamydiaceae que comprende dos géneros, Chlamydia y Chlamydophila; y las especies, Chlamydia trachomatis, C. suis, C. muridarum, Chlamydophila psittaci, C. abortus, C. felis, C. caviae, C. pecorum, y C. pneumoniae. C. psittaci causa psitacosis o clamidiosis aviar. En Argentina, los primeros casos clínicos de psitacosis fueron reportados en 1929. Los criadores de aves y quienes las poseen como mascotas, representan el grupo de mayor riesgo; pero también las personas que trabajan en pajarerías y aquellas que por su empleo se ven expuestas a contraer la enfermedad (empleados en peladeros donde se carnean y procesan pollos y otras aves para consumo, veterinarios, empleados de zoológicos, etc.). La infección en humanos se presenta como una neumonía severa; con fiebre alta, escalofríos, dolor de cabeza, mialgia y dificultad respiratoria. Ocasionalmente puede presentarse vómitos, dolor abdominal, diarrea y complicaciones como miocarditis, endocarditis, encefalitis, ictericia y fallas multiorgánicas, que pueden ser fatales sino se le administra el tratamiento adecuado. La infección en las mujeres embarazadas puede producir neumonía, hepatitis, insuficiencia renal, sepsis, parto prematuro y muerte fetal. Existen más de 465 especies de aves en las que se registró C. psittaci, incluyendo ornamentales, de corral, silvestres, acuáticas y palomas. Las patologías que pueden producir en estos animales son neumonitis, conjuntivitis, encefalomielitis, placentopatías, fetopatías, anorexia, diarrea e infecciones persistentes asintomáticas u oligosintomáticas. En bovinos, C. pecorum, C. abortus y C. psittaci producen infecciones respiratorias y genitales; que se presentan como cuadros de enteritis, artritis, encefalomielitis, endometritis e hipofertilidad. En Argentina, la infección clamidial en el ganado caprino fue asociada a daños en el tejido uterino, abortos, partos prematuros y crías débiles. En equinos, C. psittaci y C. pneumoniae producen abortos y desórdenes respiratorios, con un gran impacto en ganadería que redunda en pérdidas económicas. Considerando que existen escasos estudios eco-epidemiológicos y clínicos que reporten el estado de situación de estas infecciones en nuestro medio, es que el presente trabajo propone actualizar y profundizar el conocimiento de las especies de Clamidias de importancia médico-veterinaria presentes en la provincia de Córdoba, Argentina. El desarrollo de este proyecto aportará la implementación de técnicas que mejorarán el diagnóstico microbiológico, confirmarán los cuadros clínicos; y por lo tanto contribuirá al conocimiento de estos agentes infecciosos en nuestra región. Esta información es indispensable para los organismos responsables de la Salud Pública (Ministerios de Salud y Educación, Municipios, etc.) para que puedan obrar en consecuencia y generar sistemas de alerta temprana, tomar medidas de prevención y medidas de control frente a la presencia de un brote epidémico por alguna cepa clamidial. "Eco epidemiology of Chlamydophila psittaci, C. pecorum and C. pneumoniae: Impact on public health "The Chlamydiae are bacterial pathogens of farm animals, wildlife and pets. Besides the economic losses that occur on campuses infections of cattle, sheep, goats, swine and poultry, most species are zoonotic, infections potentially fatal can cause in humans. The order Chlamydiales is composed of the family Chlamydiaceae comprising two genera, Chlamydia and Chlamydophila, and nine species, Chlamydia trachomatis, C. suis, C. muridarum, Chlamydophila psittaci, C. abortus, C. felis, C. caviae, C. pecorum, and C. pneumoniae. The clinical manifestations in humans are associated with severe pneumonia. Also, nonspecific gastrointestinal symptoms, and complications such as myocarditis, endocarditis, encephalitis, jaundice and multiple organ failure. There are over 465 species of birds where recorded C. psittaci, including ornamental plants, poultry, wild waterfowl and pigeons. Sick birds eliminate chlamydia in all secretions, asymptomatic birds can develop the disease through contact with other infected birds. The human infection occurs by inhalation of aerosolized secretions of infected birds. The prevalence of C. psittaci records in humans, birds, horses, goats and cattle are scarce in Latin America. In Argentina it has been detected chlamydial antibody prevalence in cattle in Buenos Aires and La Pampa (5) and Rockhopper Penguins (Eudyptes chrysocome) in the town of Santa Cruz (11). Detect this pathogen in these birds and learn about potential sources of infection would be of great public health significance. To develop an efficient system of medical and veterinary surveillance is essential to have reliable diagnostic techniques for detection and identification of Chlamydia in birds, animals and humans.

The use of triphenyltetrazolium chloride in the study of dehydrogenase activity of Brucellae

Experiments for the investigation of dehydrogenase activity of washed cells of a strains of Br. abortus and another of Br. suis in presence of different single added substrates are reported. The activity was measured as the amount of formazan produced by the reduction of 2, 3, 5-triphenyltetrazolum chloride acting as a hydrogen ions acceptor, at pH 7.0. In a general manner the dehydrogenase activity of Br. suis was much more intense than that of Br. abortus (fig. 5). In the conditions of the experiments Br. abortus oxidized L-arabinose, D-galactose, D-glucose, glycerol, D-xylose, DL-alanine, D-fructose, and D-sorbitol. Brucella suis oxidized D-xylose, L-arabinose, D-glucose, D-galactose, DL-alanine, sodium acetate, maltose, glycine, D-fructose, and D-sorbitol. Glycerol was oxidized by Br. abortus but its oxidation by Br. suir was very slight. Sodium acetate and maltose were intensely oxidized by Br. suir but not by Br. abortus. The sites of more intense enzymatic acitivity were seen as small red colored round granules located in one pole of the cells.

Amplicon DNA melting analysis for the simultaneous detection of Brucella spp and Mycobacterium tuberculosis complex. Potential use in rapid differential diagnosis between extrapulmonary tuberculosis and focal complications of brucellosis.

Some sites of extrapulmonary tuberculosis and focal complications of brucellosis are very difficult to differentiate clinically, radiologically, and even histopathologically. Conventional microbiological methods for the diagnosis of extrapulmonary tuberculosis and complicated brucellosis not only lack adequate sensitivity, they are also time consuming, which could lead to an unfavourable prognosis. The aim of this work was to develop a multiplex real-time PCR assay based on SYBR Green I to simultaneously detect Brucella spp and Mycobacterium tuberculosis complex and evaluate the efficacy of the technique with different candidate genes. The IS711, bcsp31 and omp2a genes were used for the identification of Brucella spp and the IS6110, senX3-regX3 and cfp31 genes were targeted for the detection of the M. tuberculosis complex. As a result of the different combinations of primers, nine different reactions were evaluated. A test was defined as positive only when the gene combinations were capable of co-amplifying both pathogens in a single reaction tube and showed distinguishable melting temperatures for each microorganism. According to the melting analysis, only three combinations of amplicons (senX3-regX3+bcsp31, senX3-regX3+IS711 and IS6110+IS711) were visible. Detection limits of senX3-regX3+bcsp31 and senX3-regX3+IS711 were of 2 and 3 genome equivalents for M. tuberculosis complex and Brucella while for IS6110+IS711 they were of 200 and 300 genome equivalents, respectively. The three assays correctly identified all the samples, showing negative results for the control patients. The presence of multicopy elements and GC content were the components most influencing the efficiency of the test; this should be taken into account when designing a multiplex-based SYBR Green I assay. In conclusion, multiplex real time PCR assays based on the targets senX3-regX3+bcsp31 and senX3-regX3+IS711 using SYBR Green I are highly sensitive and reproducible. This may therefore be a practical approach for the rapid differential diagnosis between extrapulmonary tuberculosis and complicated brucellosis.