Brucella-associated cervical bursitis in cattle.

Bovine brucellosis poses a risk to human health and causes serious economic losses for the animal industry. This report describes the use of different diagnostic methods for the diagnosis of brucellosis in cattle affected by cervical bursitis from a slaughterhouse located in São Luís, Maranhão, Brazil. Serum samples from a total of 47 cattle with bursitis were collected and submitted to the Rose Bengal Test (RBT), and RBT-positive samples were further confirmed by the 2-mercaptoethanol (2-ME) assay. RBT indicated 85.1% (40/47) of positive samples, from which 78.7% (37/47) were confirmed by 2-ME. Immunohistochemistry detected Brucella spp. in 34.0% (16/47) of tissues with bursitis. PCR and/or bacterial isolation demonstrated that 63.8% (30/47) of samples were positive and morphologically compatible with Brucella sp. All colonies suggestive of Brucella sp. were confirmed by PCR. Isolates were further characterized by PCR Multiplex AMOS-ENHANCED, which indicated that the isolates corresponded to biovar 1, 2, 4 (43.33%). This study evidences an association between cervical bursitis and Brucella spp. infection in cattle, and that different biovars of Brucella circulate in bovine herds in Maranhão.

Inhibition of Osteoblast Function by Brucella abortus is Reversed by Dehydroepiandrosterone and Involves ERK1/2 and Estrogen Receptor.

Brucella abortus induces an inflammatory response that stimulates the endocrine system resulting in the secretion of cortisol and dehydroepiandrosterone (DHEA). Osteoarticular brucellosis is the most common presentation of the active disease in humans, and we have previously demonstrated that B. abortus infection inhibits osteoblast function. We aimed to evaluate the role of cortisol and DHEA on osteoblast during B. abortus infection. B. abortus infection induces apoptosis and inhibits osteoblast function. DHEA treatment reversed the effect of B. abortus infection on osteoblast by increasing their proliferation, inhibiting osteoblast apoptosis, and reversing the inhibitory effect of B. abortus on osteoblast differentiation and function. By contrast, cortisol increased the effect of B. abortus infection. Cortisol regulates target genes by binding to the glucocorticoid receptor (GR). B. abortus infection inhibited GRα expression. Cell responses to cortisol not only depend on GR expression but also on its intracellular bioavailability, that is, dependent on the activity of the isoenzymes 11ß-hydroxysteroid dehydrogenase (HSD) type-1, 11ß-HSD2 (which convert cortisone to cortisol and vice versa, respectively). Alterations in the expression of these isoenzymes in bone cells are associated with bone loss. B. abortus infection increased 11ß-HSD1 expression but had no effect on 11ß-HSD2. DHEA reversed the inhibitory effect induced by B. abortus infection on osteoblast matrix deposition in an estrogen receptor- and ERK1/2-dependent manner. We conclude that DHEA intervention improves osteoblast function during B. abortus infection making it a potential candidate to ameliorate the osteoarticular symptoms of brucellosis.

Padronização e validação do ELISA indireto para diagnostico de brucelose bovina utilizando como antigeno lipopolissacarideo e extrato protéico solúvel de Brucella abortus

A brucelose é uma zoonose de distribuição mundial, acarreta problemas sanitários importantes e prejuízos econômicos vultuosos a pecuária. A vacinação de fêmeas bovinas de 3 a 8 meses de idade e o sacrifício obrigatório de animais sororreagentes são importantes medidas de controle desta doença, descritas sob forma de lei através do Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose (PNCEBT). Visto a obrigatoriedade da eliminação dos animais sororreagentes e a inexistência de um teste preciso para detecção da doença, objetivou-se padronizar e validar dois ensaios imunoenzimáticos indiretos (iELISA) para o diagnóstico da brucelose bovina, capaz de diferenciar animal recém-vacinado de naturalmente infectado, utilizando como antígeno Lipopolissacarideo (LPS) e Extrato Protéico Solúvel (EPS), ambos extraídos da cepa 1119-3 de Brucella abortus. Para padronização utilizou-se duas concentrações dos antígenos; diferentes diluições de um pool de amostras negativas para brucelose, positivas de fêmeas bovinas recém-vacinadas contra brucelose e outro de fêmeas bovinas naturalmente infectadas para brucelose; e duas diluições de anticorpo secundário. Após serem padronizados, os testes foram validados com 96 amostras de soros sanguíneos de bovinos previamente testados pelo Antígeno Acidificado Tamponado (AAT) e 2-Mercaptoetanol (2-ME), sendo 64 positivas (fêmeas bovinas naturalmente infectadas e recém-vacinadas) e 32 negativas. Os cálculos estatísticos foram feitos separando-se os grupos de amostras positivas de fêmeas bovinas naturalmente infectadas e de fêmeas bovinas recém-vacinadas. Para comparação entre os resultados dos testes foi usado o coeficiente kappa. Calculou-se a sensibilidade e a especificidade em relação ao 2-ME. Os testes iELISA/LPS e iELISA/EPS para amostras de soro de fêmeas bovinas naturalmente infectadas, e iELISA/LPS para amostras de soros de fêmeas bovinas recém-vacinadas, tiveram alta sensibilidade e especificidade, e concordância excelente com o 2-ME, porém não foi capaz de diferenciar animais recém-vacinados de naturalmente infectados, ao contrário do iELISA/EPS para amostras de soros de fêmeas bovinas recém-vacinadas, capaz de fazer esta diferenciação, porém com baixa sensibilidade e fraca concordância com o 2-ME. Constatou-se que o EPS pode se tornar um potencial antígeno para testes diagnósticos de brucelose.

The malasseziose is a major disease causing inflammation of the hearing of dogs and cats. Thus, the objective was to report the cases of a cat and a dog assisted in the Veterinary Hospital of the Federal University of Uberlândia, and diagnosed with otitis externa by Malassezia sp. from laboratory and subjected to treatment with Auritop®.

Brucelose bovina no Estado de Roraima: estudo retrospectivo / Bovine brucellosis in roraima state: retrospective study

O objetivo do presente trabalho foi determinar a frequência de propriedades positivas (focos) e de fêmeas bovinas soropositivas para a brucelose no Estado de Roraima, bem como identificar fatores de risco. Foram utilizados dados da Agência de Defesa Agropecuária do Estado de Roraima (ADERR), coletados de suas quatro microrregiões, durante o período de janeiro de 2007 a julho de 2009. Durante esse período, foram examinadas 387 propriedades, e 9.087 soros de fêmeas bovinas com idade ≥ 24 meses foram submetidos ao diagnóstico sorológico da brucelose. Para a análise de fatores de risco, foi utilizada a regressão logística múltipla. Uma propriedade foi considerada foco quando apresentou pelo menos um animal soropositivo. Das propriedades investigadas, 106 (27,4%) apresentaram pelo menos um animal soropositivo e, dos animais analisados, 369 (4,1%) foram soropositivos. Os fatores de risco identificados foram: possuir mais de 51 animais no rebanho(odds ratio = 1,87; p = 0,015) e alugar pasto (odds ratio = 2,20; p = 0,003). Sugere-se que esforços sejam concentrados na intensificação da vacinação de bezerras no Estado de Roraima com o intuito de que seja alcançada uma prevalência compatível com as ações de teste e sacrifício de animais.

BOVINE BRUCELLOSIS IN RORAIMA STATE: RETROSPECTIVE STUDY. The aim of this study was to determine the frequency of positive herds (foci) and seropositive bovine females for brucellosis in the state of Roraima, Northern region of Brazil, as well as to identify risk factors. Data from the Agency of Agricultural Protection in the state, collected from its four microregions during the January 2007 to July 2009 period, were used. During this period, 387 herds were examined, and 9,087 adult bovine female sera were submitted to the serological diagnosis of brucellosis. For risk factor analysis the multiple logistic regression was used. A herd was considered focus when presented at least one seropositive animal. Of the herds investigated, 106 (27.4%) had at least one seropositive animal, and of the animals examined, 369 (4.1%) were seropositive. Risk factors identified were: herd size larger than 51 animals (odds ratio = 1.87; p = 0.015) and pasture rental (odds ratio = 2.20; p = 0.003). It is suggested that efforts must be concentrated on the intensification of heifer vaccination in the State of Roraima in order to reach a prevalence value compatible with actions of test and sacrifice of animals.

Brucelose bovina no Estado de Roraima: estudo retrospectivo / Bovine brucellosis in roraima state: retrospective study

O objetivo do presente trabalho foi determinar a frequência de propriedades positivas (focos) e de fêmeas bovinas soropositivas para a brucelose no Estado de Roraima, bem como identificar fatores de risco. Foram utilizados dados da Agência de Defesa Agropecuária do Estado de Roraima (ADERR), coletados de suas quatro microrregiões, durante o período de janeiro de 2007 a julho de 2009. Durante esse período, foram examinadas 387 propriedades, e 9.087 soros de fêmeas bovinas com idade ≥ 24 meses foram submetidos ao diagnóstico sorológico da brucelose. Para a análise de fatores de risco, foi utilizada a regressão logística múltipla. Uma propriedade foi considerada foco quando apresentou pelo menos um animal soropositivo. Das propriedades investigadas, 106 (27,4%) apresentaram pelo menos um animal soropositivo e, dos animais analisados, 369 (4,1%) foram soropositivos. Os fatores de risco identificados foram: possuir mais de 51 animais no rebanho(odds ratio = 1,87; p = 0,015) e alugar pasto (odds ratio = 2,20; p = 0,003). Sugere-se que esforços sejam concentrados na intensificação da vacinação de bezerras no Estado de Roraima com o intuito de que seja alcançada uma prevalência compatível com as ações de teste e sacrifício de animais. (AU)

BOVINE BRUCELLOSIS IN RORAIMA STATE: RETROSPECTIVE STUDY. The aim of this study was to determine the frequency of positive herds (foci) and seropositive bovine females for brucellosis in the state of Roraima, Northern region of Brazil, as well as to identify risk factors. Data from the Agency of Agricultural Protection in the state, collected from its four microregions during the January 2007 to July 2009 period, were used. During this period, 387 herds were examined, and 9,087 adult bovine female sera were submitted to the serological diagnosis of brucellosis. For risk factor analysis the multiple logistic regression was used. A herd was considered focus when presented at least one seropositive animal. Of the herds investigated, 106 (27.4%) had at least one seropositive animal, and of the animals examined, 369 (4.1%) were seropositive. Risk factors identified were: herd size larger than 51 animals (odds ratio = 1.87; p = 0.015) and pasture rental (odds ratio = 2.20; p = 0.003). It is suggested that efforts must be concentrated on the intensification of heifer vaccination in the State of Roraima in order to reach a prevalence value compatible with actions of test and sacrifice of animals.(AU)

Padronização e validação do ELISA indireto para diagnostico de brucelose bovina utilizando como antigeno lipopolissacarideo e extrato protéico solúvel de Brucella abortus

A brucelose é uma zoonose de distribuição mundial, acarreta problemas sanitários importantes e prejuízos econômicos vultuosos a pecuária. A vacinação de fêmeas bovinas de 3 a 8 meses de idade e o sacrifício obrigatório de animais sororreagentes são importantes medidas de controle desta doença, descritas sob forma de lei através do Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose (PNCEBT). Visto a obrigatoriedade da eliminação dos animais sororreagentes e a inexistência de um teste preciso para detecção da doença, objetivou-se padronizar e validar dois ensaios imunoenzimáticos indiretos (iELISA) para o diagnóstico da brucelose bovina, capaz de diferenciar animal recém-vacinado de naturalmente infectado, utilizando como antígeno Lipopolissacarideo (LPS) e Extrato Protéico Solúvel (EPS), ambos extraídos da cepa 1119-3 de Brucella abortus. Para padronização utilizou-se duas concentrações dos antígenos; diferentes diluições de um pool de amostras negativas para brucelose, positivas de fêmeas bovinas recém-vacinadas contra brucelose e outro de fêmeas bovinas naturalmente infectadas para brucelose; e duas diluições de anticorpo secundário. Após serem padronizados, os testes foram validados com 96 amostras de soros sanguíneos de bovinos previamente testados pelo Antígeno Acidificado Tamponado (AAT) e 2-Mercaptoetanol (2-ME), sendo 64 positivas (fêmeas bovinas naturalmente infectadas e recém-vacinadas) e 32 negativas. Os cálculos estatísticos foram feitos separando-se os grupos de amostras positivas de fêmeas bovinas naturalmente infectadas e de fêmeas bovinas recém-vacinadas. Para comparação entre os resultados dos testes foi usado o coeficiente kappa. Calculou-se a sensibilidade e a especificidade em relação ao 2-ME. Os testes iELISA/LPS e iELISA/EPS para amostras de soro de fêmeas bovinas naturalmente infectadas, e iELISA/LPS para amostras de soros de fêmeas bovinas recém-vacinadas, tiveram alta sensibilidade e especificidade, e concordância excelente com o 2-ME, porém não foi capaz de diferenciar animais recém-vacinados de naturalmente infectados, ao contrário do iELISA/EPS para amostras de soros de fêmeas bovinas recém-vacinadas, capaz de fazer esta diferenciação, porém com baixa sensibilidade e fraca concordância com o 2-ME. Constatou-se que o EPS pode se tornar um potencial antígeno para testes diagnósticos de brucelose.(AU)

The malasseziose is a major disease causing inflammation of the hearing of dogs and cats. Thus, the objective was to report the cases of a cat and a dog assisted in the Veterinary Hospital of the Federal University of Uberlândia, and diagnosed with otitis externa by Malassezia sp. from laboratory and subjected to treatment with Auritop®.(AU)

Virulence of Brucella abortus isolated from cattle and water buffalo.

Brucellosis has been documented in domestic water buffalo (Bubalus bubalis) but published literature is limited despite the importance of this species in tropical agricultural systems. The objective of this study was to compare the virulence of Brucella abortus isolates recovered from cattle and water buffalo. Nineteen strains of B. abortus from cattle and domestic water buffalo in Trinidad were intraperitoneally inoculated into BALB/c mice. Spleens were cultured for B. abortus and histopathological severity scores were calculated based on lymphoid depletion, lymphoid necrosis, splenitis, and macrophage accumulation. A general linear model approach was used to estimate the effect of isolate source (cattle versus water buffalo) on virulence. Isolates of water buffalo origin were significantly less virulent in the mouse model based on recovered B. abortus from splenic tissues, spleen/weight ratio, and lymphoid necrosis but not overall histopathological severity scores. Further investigation of isolates recovered from water buffalo might provide the key to the development of procedures for brucellosis control in tropical environments.

Comparative study on responses of cattle and water buffalo (Bubalus bubalis) to experimental inoculation of Brucella abortus biovar 1 by the intraconjunctival route--a preliminary report.

The preliminary study was conducted to assess the virulence of a strain of Brucella abortus (1969D) and to compare the susceptibility of water buffalo and cattle calves to infection by the intraconjunctival route. Seven of each cattle and water buffalo (Bubalus bubalis) calves aged 3-6 months were inoculated intraconjunctivally with counts ranging from 1.5 × 10(7) to 1.7 × 10(10) colony forming units of B. abortus. Animals were monitored over an 8-week period for clinical manifestations and serological and hematological evidence of infection. At slaughter, eight lymph nodes from each animal were sampled for bacteriological and histopathological assessments. Lymph nodes from three water buffalo (43%) and five cattle (71%) yielded B. abortus (P=0.048). Parotid/prescapular lymph nodes were most sensitive in detecting B. abortus. Our data suggest that B. abortus strain 1969D may be used as challenge strain, and water buffalo appeared to have a lower susceptibility to B. abortus infection than cattle.

Pathogenesis of bovine brucellosis.

Bovine brucellosis is one of the most important zoonotic diseases worldwide, and is of particular significance in developing countries. The disease, which results in serious economic losses due to late term abortion, stillborn and weakly calves, is caused by Gram negative coccobacilli bacteria of the genus Brucella. Lesions consist of necrotic placentitis and interstitial mastitis in pregnant cows, and fibrinous pleuritis with interstitial pneumonia in aborted fetuses and newborn calves. This article considers the pathogenesis of Brucella abortus and reviews the ability of the pathogen to invade phagocytic and non-phagocytic host cells, resist the acidified intraphagosomal environment, and inhibit phagosome-lysosome fusion. Significant aspects of innate and adaptive immunity against brucellosis are also discussed.

Pathological, immunohistochemical and bacteriological study of tissues and milk of cows and fetuses experimentally infected with Brucella abortus.

This report describes a pathological, immunohistochemical and bacteriological study of 42 cows and their progeny (aborted fetuses, weak premature calves, and healthy full-term calves) infected at 6-7 months of gestation by conjunctival inoculation with Brucella abortus. Samples were collected at necropsy within 48 h of abortion or parturition. The most significant lesions were necrotizing and suppurative placentitis and lymphohistiocytic mastitis in cows, and fibrinous pleuritis, fibrinous pericarditis and bronchopneumonia in aborted fetuses. B. abortus was isolated more frequently from milk samples than from mammary tissues, and milk samples from cows with mastitis were often infected. Organisms were often demonstrated immunohistochemically and by culture in tissues showing moderate to severe histological changes.