Seroprevalencia de leptospirosis y brucelosis en búfalos de agua (Bubalus bubalis) en Tierralta Córdoba, Colombia

En la producción zootécnica, la brucelosis y la leptospirosis ocasionan problemas reproductivos y son una limitante en salud y en producción animal. El objetivo fue determinar la presencia de anticuerpos contra Leptospira spp. y Brucella abortus, en una población bufalera, en el municipio de Tierralta, Córdoba. Se realizó un estudio descriptivo de corte transversal, que incluyó un total de 144 búfalos de la raza Murrah, destinados al doble propósito. Para el diagnóstico de Leptospira spp., se implementó la prueba de aglutinación microscópica, con 13 serogrupos, 19 serovares pertenecientes a 5 especies de Leptospira patógenas y para brucelosis Rosa de Bengala y C-Elisa. La seroprevalencia para Leptospira spp. fue del 87,5 %, el serogrupo Mini fue el de mayor frecuencia, pero Grippotyphosa presentó el mayor título. El 16,67 % de los búfalos evaluados presentaron títulos iguales o superiores a 1:800, asociados con infección actual o reciente. La alta seroprevalencia, se puede deber a las características ambientales de la zona, que brinda las condiciones favorables para el crecimiento y el mantenimiento de este patógeno que, sumado al comportamiento de los búfalos de revolcarse, los hace propensos a las infecciones con bacterias del género Leptospira sp., porque a menudo, las fuentes de agua están contaminadas por este patógeno. La seroprevalencia contra B. abortus por Rosa de Bengala y Elisa-C fue del 2,08 %. Todos los títulos determinados corresponden a procesos infecciosos. No hubo signos clínicos de enfermedad y la carencia de registros productivos no permitió determinar el efecto sobre los parámetros reproductivos.

In animal production, brucellosis and leptospirosis cause reproductive problems and limit animal health and production. The objective was to determine the presence of antibodies against Leptospira spp. and Brucella abortus in a buffalo population in the municipality of Tierralta, Cordoba. A descriptive, transversal study was carried out including a population of 144 Murrah´s breed buffalos destined for beef and milk production. For the Leptospira spp. diagnostic, was used a rapid slide agglutination test with 13 serogrups and 19 serovars belonging to 5 pathogenic Leptospira species was implemented and for Brucella abortus Rose Bengal and C-Elisa was used. Seroprevalence of Leptospira spp. was 87,5 %, serogrup Mini was the most frequent, but Grippotyphosa showed the higher titer. The 16,67 % of the buffaloes evaluated presented titles equal to or above than 1:800 associated with current or recent infection. High seroprevalence may be due to environmental characteristics of the zone, which gives favorable conditions for the growth and maintenance of this pathogen, these factors in conjunction with the habit of wallowing makes them prone to suffering infections caused by bacteria of the genera Leptospira sp. since water sources are often contaminated. Seroprevalence against B. abortus by Rose-Bengal and C-Elisa was 2,08 %, the determined titers correspond to infectious processes. There were no clinical manifestations of disease and the effects on reproductive parameters were not determined because of the lack of productive registries.

Presence of Chlamydia abortus in colostrum, milk, and vaginal discharge samples of sheep

Background: The main transmission route of Chlamydia abortus is by ingesting the microorganism that has been eliminated in vaginal secretions, placental membranes or abortions that contaminate the environment and, possibly, through milk and colostrum. Elimination through vaginal secretions is well documented. However, there are no reports about isolation and identification of C. abortus in the colostrum or milk of infected sheep, so it is important to determine whether or not C. abortus may be present in these secretions, which are the only food of lambs. Objective: To detect C. abortus in colostrum, milk, and vaginal secretions of sheep with a history of reproductive disorders. Methods: Colostrum, milk, and vaginal exudates were collected from 66 sheep. The samples were inoculated in mouse fibroblast cell cultures and the presence of C. abortus determined by direct immunofluorescence. Results: 19 out of 66 colostrum samples (28.7%), 14 out of 66 milk samples (21.2%) and 17 out of 66 vaginal swabs (25.7%) were positive for C. abortus. The 50 samples positive for isolation and detected by immunofluorescence, together with 42 negative samples were subjected to qPCR to amplify a fragment of the ompA gene from C. abortus. Thirty-eight of the 92 samples processed by this technique were positive for C. abortus. Conclusion: The results demonstrated the presence of C. abortus in a high proportion in colostrum, milk and vaginal secretions of infected sheep. To the best of our knowledge, this is the first field study confirming the presence of C. abortus in colostrum, which shows that excretion of Chlamydia by lactogenesis could occur in the first hours after birth.

Antecedentes: La principal vía de transmisión de C. abortus es la ingestión del microorganismo que ha sido eliminado en las secreciones vaginales, membranas placentarias, abortos y, posiblemente, a través de la leche y el calostro. La eliminación a través de secreciones vaginales está bien documentada. Sin embargo, no existen reportes del aislamiento e identificación de C. abortus en el calostro o la leche de ovejas infectadas, por lo que es importante determinar si la bacteria puede o no estar presente en estas secreciones, que son el único alimento de los corderos. Objetivo: Detectar la presencia de C. abortus in calostro, leche y secreciones vaginales de ovejas con antecedentes de problemas reproductivos. Método: Con el propósito de aislar e identificar C. abortus en estas secreciones, se recolectó calostro, leche y exudado vaginal de 66 ovejas. Las muestras fueron inoculadas en cultivos celulares de fibroblastos de ratón y se determinó la presencia de la bacteria por inmunofluorescencia directa. Resultados: Fueron positivas 19 de 66 muestras de calostro (28,7%), 14 de 66 muestras de leche (21,2%) y 17 de 66 hisopos vaginales (25,7%). Las 50 muestras positivas al aislamiento y detectadas por inmunofluorescencia, junto con 42 negativas se sometieron a qPCR para amplificar un fragmento del gen ompA de C. abortus; 38 de las 92 muestras procesadas por esta técnica fueron positivas para C. abortus. Conclusión: Los resultados del presente estudio demostraron la presencia de C. abortus en una alta proporción en el calostro, la leche y las secreciones vaginales de ovejas infectadas. Este es el primer estudio de campo que confirma la presencia de C. abortus en calostro, lo que demuestra que la excreción de clamidia por lactogénesis podría ocurrir en las primeras horas después del nacimiento.

Antecedentes: A principal via de transmissão da Chlamydia abortus é a ingestão do microrganismo que foi eliminado nas secreções vaginais, membranas placentárias ou abortos que contaminam o meio ambiente e, possivelmente, através do leite e colostro. A eliminação pelas secreções vaginais está bem documentada. No entanto, não há relatos de isolamento e identificação de C. Abortus no colostro ou leite de ovelhas infectadas, por isso é importante verificar se a bactéria pode estar ou não presente nessas secreções, único alimento dos cordeiros. Objetivo: Detectar a presença de C. Abortus no colostro, leite e secreções vaginais de ovelhas com histórico de distúrbios reprodutivos Métodos: Para isolar e identificar C. Abortus nessas secreções, foram coletados colostro, leite e exsudato vaginal de 66 ovelhas. As amostras foram inoculadas em cultura de células de fibroblastos de camundongo e a presença da bactéria determinada por imunofluorescência direta. Resultados: 19 de 66 amostras de colostro (28,7%), 14 de 66 amostras de leite (21,2%) e 17 de 66 esfregaços vaginais (25,7%) sendo positivos. As 50 amostras positivas para isolamento e detectadas por imunofluorescência, juntamente com as 42 negativas, foram submetidas a qPCR para amplificar um fragmento do gene ompA de C. Abortus. Trinta e oito das 92 amostras processadas por esta técnica foram positivas para C. Abortus. Conclusão: Os resultados do presente estudo demonstraram a presença de C. Abortus em alta proporção no colostro, leite e secreções vaginais de ovelhas infectadas. Este trabalho é o primeiro estudo de campo na literatura científica confirmando a presença de C. Abortus no colostro, o que mostra que a excreção da clamídia por lactogênese pode ocorrer nas primeiras horas após o nascimento.

Detección de Chlamydia spp. en leucocitos de sangre periférica de ovinos sin aborto previo / Detection of Chlamydia spp. In Peripheral Blood Leukocytes in Ewes without any Previous Abortion

Resumen El aborto enzootico ovino es una enfermedad causada por Chlamydia abortus. Es considerada una zoonosis y una de las principales causas de pérdidas económicas en estas explotaciones. Este trabajo se enfocó en utilizar el cultivo de leucocitos de animales sin signos de abortos y la detección de anticuerpos para determinar la posible presencia de C. abortus en explotaciones de traspatio. Se obtuvieron 42 muestras de sangre periférica de ovejas de diferentes poblaciones. La detección de Chlamydia abortus se realizó mediante la tinción de Giemsa y la técnica de PCR. La detección de anticuerpos anti-C. abortus se dio mediante una técnica de ELISA comercial. Los resultados mostraron 21 muestras positivas mediante la técnica de PCR, de las cuales solo 10 fueron positivas mediante la técnica de Giemsa, mientras que 22 sueros mostraron anticuerpos anti-C. abortus. En este estudio el 38,1 % de las muestras fueron positivas a la infección por C. abortus, como se confirmó mediante PCR y serología. En conclusión, los leucocitos de sangre periférica pueden ser útiles para detectar una infección por Chlamydia spp. en explotaciones sin historial de abortos, con lo que se puede conocer la prevalencia real del aborto enzootico ovino en México.

Abstract The Ewes Enzootic Abortion is a disease caused byChlamydia abortus. It is deemed a zoonosis and one of the leading causes of financial losses in this type of business. This article focuses on using the culture of leukocytes from animals without any abortion symptoms and antibody detection to determine the potential presence ofC. abortusin backyard exploitations. Forty-two samples of peripheral blood were obtained from ewes in different populations. The detection ofChlamydia abortuswas carried out by using the Giemsa dye and PCR technique. Anti-C. Abortusantibody detection was performed through a commercial ELISA technique. Results showed 21 positive samples using the PCR, and only ten were positive according to the Giemsa dye, while 22 serum samples showed anti-C. abortusantibody. In this study, 38.1% of the samples were positive for theC. abortusinfection, as verified with the PCR and serology. In conclusion, peripheral blood leukocytes can be helpful to detect an infection caused byChlamydiaspp. Animal exploitation without any previous abortion allows knowing the real prevalence of ewes' enzootic abortion in Mexico.

Seropositividad de Brucella abortus en ganado bovino de Aguazul- Casanare / Seropositivity of Brucella abortus in cattle from Aguazul- Casanare

Resumen Introducción. La Brucelosis bovina, es una enfermedad bacteriana e infecto contagiosa, causada por Brucella abortus. Se transmite a través de la ingestión de forrajes y aguas contaminadas con descargas vaginales infectadas, que conlleva a una patología del sistema reproductor en bovinos, que impactan la sanidad pecuaria y la economía de la agroindustria. Objetivo. Evaluar el comportamiento de la brucelosis bovina en el municipio de Aguazul, Casanare (Colombia) y los factores asociados al desarrollo de esta enfermedad. Metodología. Estudio descritptivo de 26.187 muestras de suero sanguíneo de ganado bovino evaluadas, que corresponden a 260 predios del municipio de Aguazul. Se empleó técnicas serológicascomo rosa de bengala, fluorescencia polarizada y ELISA competitiva para la evaluación de positividad a Brucella abortus y se evaluaron las pérdidas económicas asociadas a positividad en los ensayos de laboratorio. Resultados y conclusiones. La positividad a brucelosis bovina correspondió al 1%, que corresponde a hembras menores de 24 meses de edad y entre 37 a 48 meses y, machos entre los 57 a 68 meses de edad. Se sugiere consolidar esfuerzos en investigación para evaluar los factores que contribuyen a la seropositividad en el ganado y el riesgo para la propagación y mantenimiento de la enfermedad.

Abstract Introduction. Bovine brucellosis is a contagious bacterial and infectious disease caused by Brucella abortus. It is transmitted through the ingestion of forage and water contaminatedwith infected vaginal discharges, which leads to a pathology of the reproductive system inbovines, which impacts livestock health and the economy of agribusiness. Aim. To evaluatethe behavior of bovine brucellosis in the municipality of Aguazul, Casanare (Colombia) and the factors associated with the development of this disease. Methodology. Descriptive study of 26,187 bovine blood serum samples evaluated, corresponding to 260 farms in the municipality of Aguazul. Serological techniques such as rose bengal, polarized fluorescence, and competitive ELISA were used to evaluate positivity to Brucella abortus and the economic losses associated with positivity in laboratory tests were evaluated. Results and conclusions. The positivity to bovine brucellosis corresponded to 1%, which corresponds to females under 24 months of age and between 37 to 48 months and males between 57 to 68 months of age. It is suggested to consolidate research efforts to evaluate the factors that contribute to seropositivity in cattle and the risk for the spread and maintenance of the disease

Inquérito sorológico de brucelose em machos e fêmeas bovinas em fazendas do Estado de Mato Grosso do Sul / Serological survey of brucellosis in males and females of cattle in the State of Mato Grosso do Sul

Objetivou-se avaliar a ocorrência de animais soro reagentes à brucelose bovina em fazendas localizadas no Estado de Mato Grosso do Sul, por meio de exame sorológico utilizando o Antígeno Acidificado Tamponado (AAT) e discutir as possíveis diferenças entre as soroprevalências de fêmeas e machos. Foram avaliados, a partir do teste de triagem com Antígeno Acidificado Tamponado (AAT), 724 bovinos da raça Nelore, sendo 274 machos e 450 fêmeas, provenientes de oito propriedades com histórico de problemas reprodutivos. O teste foi procedido conforme o protocolo determinado pelo Ministério da Agricultura, Pecuária e Abastecimento (MAPA). Os resultados demonstraram baixa soroprevalência da doença nos bovinos testados, sendo detectada prevalência para a doença de 1,10% nos machos e 2,88% nas fêmeas.Quando se considera o touro isoladamente nos rebanhos, pode-se perceber que a fertilidade é muito mais importante nos machos do que nas fêmeas individualmente, uma vez que os touros podem se acasalar com um número muito maior de fêmeas, seja na monta natural ou na inseminação artificial, demonstrando a importância do inquérito epidemiológico na população geral, principalmente nos machos. A maior frequência da doença foi encontrada nas fêmeas podendo estar relacionada à infecção por Brucella spp. no ambiente decorrente de parto ou aborto tornando as fêmeas transmissoras permanentes da doença.

The objective of this study was to evaluate the occurrence of seroreactive animals to bovine brucellosis in farms located in the State of Mato Grosso do Sul, by means of a serological examination using the Acidified Buffered Antigen (AAT) and to discuss the possible differences between the seroprevalence of females and males. A total of 724 Nellore cattle, 274 males and 450 females, from eight farms with a history of reproductive problems, were evaluated using the screening test with Acidified Buffered Antigen (AAT). The test was carried out according to the protocol determined by the Ministry of Agriculture, Livestock and Food Supply (MAPA).The results showed a low seroprevalence of the disease in the tested cattle, with a prevalence of 1.10% in males and 2.88% in females.When considered the bull alone in herds, it can be shown that fertility is much more important in males than in females individually, since bulls can mate with a much larger number of females, either in natural mating or in artificial insemination, demonstrating the importance of epidemiological survey in the general population, especially in males. The highest frequency of the disease was found in females and may be related to infection by Brucella spp. in the environment from childbirth or abortion making females permanent transmitters of the disease.

Inquérito sorológico de brucelose em machos e fêmeas bovinas em fazendas do Estado de Mato Grosso do Sul / Serological survey of brucellosis in males and females of cattle in the State of Mato Grosso do Sul

Objetivou-se avaliar a ocorrência de animais soro reagentes à brucelose bovina em fazendas localizadas no Estado de Mato Grosso do Sul, por meio de exame sorológico utilizando o Antígeno Acidificado Tamponado (AAT) e discutir as possíveis diferenças entre as soroprevalências de fêmeas e machos. Foram avaliados, a partir do teste de triagem com Antígeno Acidificado Tamponado (AAT), 724 bovinos da raça Nelore, sendo 274 machos e 450 fêmeas, provenientes de oito propriedades com histórico de problemas reprodutivos. O teste foi procedido conforme o protocolo determinado pelo Ministério da Agricultura, Pecuária e Abastecimento (MAPA). Os resultados demonstraram baixa soroprevalência da doença nos bovinos testados, sendo detectada prevalência para a doença de 1,10% nos machos e 2,88% nas fêmeas.Quando se considera o touro isoladamente nos rebanhos, pode-se perceber que a fertilidade é muito mais importante nos machos do que nas fêmeas individualmente, uma vez que os touros podem se acasalar com um número muito maior de fêmeas, seja na monta natural ou na inseminação artificial, demonstrando a importância do inquérito epidemiológico na população geral, principalmente nos machos. A maior frequência da doença foi encontrada nas fêmeas podendo estar relacionada à infecção por Brucella spp. no ambiente decorrente de parto ou aborto tornando as fêmeas transmissoras permanentes da doença.

The objective of this study was to evaluate the occurrence of seroreactive animals to bovine brucellosis in farms located in the State of Mato Grosso do Sul, by means of a serological examination using the Acidified Buffered Antigen (AAT) and to discuss the possible differences between the seroprevalence of females and males. A total of 724 Nellore cattle, 274 males and 450 females, from eight farms with a history of reproductive problems, were evaluated using the screening test with Acidified Buffered Antigen (AAT). The test was carried out according to the protocol determined by the Ministry of Agriculture, Livestock and Food Supply (MAPA).The results showed a low seroprevalence of the disease in the tested cattle, with a prevalence of 1.10% in males and 2.88% in females.When considered the bull alone in herds, it can be shown that fertility is much more important in males than in females individually, since bulls can mate with a much larger number of females, either in natural mating or in artificial insemination, demonstrating the importance of epidemiological survey in the general population, especially in males. The highest frequency of the disease was found in females and may be related to infection by Brucella spp. in the environment from childbirth or abortion making females permanent transmitters of the disease.

Zoonotic smooth and rough Brucella in dogs: seroprevalence and associated factors in an Atlantic Rainforest area of the state of Paraíba, Northeastern Brazil / Brucella zoonóticalisa e rugosa em cães: soroprevalência e fatores associados em área de Mata Atlântica do estado da Paraíba, Nordeste brasileiro

ABSTRACT: Canine brucellosisis an infectious disease caused by bacteria of the genus Brucella, with world wide distribution and zoonotic impact, and in humans and animals is a neglected disease. In the present study, the sero prevalence of B. canis and B. abortus were determined in a probabilistic sample of housed dogs from the Atlantic Rainforest area of the state of Paraíba, Brazil, and the factors associated with sero positivity. A total of 386 dogs over three months of age were used. For the search for anti-B.canis antibodies the agar gel immune diffusion test (IDGA) was used as a screening and IDGA+2ME as confirmatory test, and to search for anti-B. abortus antibodies the Rose Bengal test (RBT) test was used. Apparent and real prevalences were calculated, and robust Poisson regression was used to identify factors associated with prevalence. The real prevalence fB. Canis was 12.6% and of B. abortus was 22.8%. The factors associated with sero positivity for B. canis were age greater than 10 years (prevalence ratio; PR = 6.38; P = 0.024) and dogs reared in they ard (PR = 5.20; P = 0.035) and for B. abortus was no treplacement of water of animals everyday (PR = 1.48; P = 0.033). It can be concluded that the prevalence of B. canis and B. Abortus in the region is high, which warns to the adopting of control and prevention measures, as well as greater care in the management of animals, especially for elderly dogs.

RESUMO: A brucelose canina é uma doença infecciosa causada por bactérias do gênero Brucella, com distribuição mundial e de caráter zoonótico, e em humanos e animais é uma doença negligenciada. No presente estudo foram determinados as soroprevalências de B. canis e B. abortusem uma amostra probabilística de cães domiciliados da área urbana de oito municípios localizados na região da Mata Atlântica do Estado da Paraíba, Brasil, e os fatores associados com a soropositividade. Foram utilizados 386 cães com mais de três meses de idade. Para a pesquisa de anticorpos anti-B. canis foi utilizado o teste de imunodifusão em gel de ágar (IDGA) como triagem e IDGA+2ME como confirmatório, e para a pesquisa de anticorpos anti-B. abortusfoi utilizado o teste do antígeno acidificado tamponado (AAT). Foram calculadas as prevalências aparente e real, e para a identificação de fatores associados com a prevalência foi empregada regressão robusta de Poisson. A prevalência real de B. canis foi de 12,6% e de B. abortusfoi 22,8%. Os fatores associados com a soropositividade para B. canis foram idade maior que 10 anos (razão de prevalência; RP = 6,38; P = 0,024) e cães criados presos no quintal (RP = 5,20; P = 0,035) e para B. abortus foi não trocar a água dos animais todos os dias (RP = 1,48; P = 0,033). Conclui-se que a prevalência de B. canis e B. abortusem cães da região é alta, o que alerta para a necessidade de adoção de medidas de controle e prevenção, bem como são sugeridos maiores cuidados no manejo dos animais, sobretudo cães idosos.

Phylogeny of Brucella abortus strains isolated in the Russian Federation

Objective: To study Brucella (B.) abortus strains isolated in the Russian Federation, in order to identify their detailed position in the phylogenetic structure of the species global population as well as to determine genetic relationships for isolates from different geographical areas. Methods: Based on Bayesian method, the whole genome single-nucleotide polymorphism (SNP) analysis of 258 B. abortus strains from different geographical areas of the world including 20 B. abortus strains isolated in Russia was carried out. Results: The core genome SNP analysis of the B. abortus isolates allowed describing the main genetic lineages. The Russian strains entered two separate clades, including the basal branch and the C1 branch that is widely spread in Eurasia. The data on the isolation time was used for the dating of phylogenetic tree, and also the estimated time frame for the B. abortus genotype diversification was determined. There were sets of specific SNPs identified, which defined each of the genotypes and sub-genotypes. Conclusions: A significant genetic diversity of the brucellosis pathogen strains from Russia has been proven. The sets of unique specific SNPs described in our study may become one of the elements within a bio-informational analysis algorithm to be used for epidemiological study of brucellosis outbreaks, including those caused by new (atypical) genetic variants of B. abortus.

Seroprevalence and Risk Factors of Brucellosis in Dogs in Enugu and Anambra States, Nigeria / Seroprevalencia y factores de riesgo de la brucelosis en perros de los Estados Enugu y Anambra, Nigeria

Abstract Seroprevalence of the antibodies of Brucella canis and Brucella abortus in dogs was assessed using a cross-sectional survey in Anambra and Enugu States, Nigeria. A total of 123 Companion dogs made up of 65 clinic dogs, 34 slaughter dogs and 24 household dogs were screened. For B. abortus antibody assay, the collected serum was used for Rose Bengal plate test (RBPT), Serum agglutination test (SAT) and Solid Phase Immunoassay technique with Immunocomb® Canine Brucellosis Antibody Test Kit was used. Out of the 123 dogs screened, none was positive for Brucella abortus antibodies while 34 (27.7%) of the dogs screened were positive for B. canis antibodies. There was a significant association (P<0.05) between infection and sex, the infection was significantly higher (P<0.05) in female than male dogs. Prevalence was significantly higher (P<0.05) in Exotic breeds than in mixed and local dog breeds. There was no association (P>0.05) between infection and antibody titre levels in the different categories of dogs. However, there was significant association (P<0.05) between the presence of Brucella canis antibodies and free roaming of dogs. This study provides the first serological evidence of B. canis infection in dogs in Enugu and Anambra States. This shows that B. canis is endemic in both states, underscoring the need for further studies. Female dogs, exotic breeds and freely roaming dogs are at a higher risk of Brucella infection in the study area; therefore, preventive and control measures are strongly recommended.

Resumen Se evaluó la seroprevalencia de los anticuerpos de Brucella canis y Brucella abortus en perros usando un sondeo transversal en los Estados Anambra and Enugu, Nigeria. Se examinó un total de 123 perros de compañía, de los cuales 65 eran perros de clínica, 34 perros de matadero y 24 perros caseros. Para el ensayo de anticuerpos de B. abortus, el suero muestreado se usó para la prueba de Rosa de Bengala (RBPT), prueba de aglutinación del suero (SAT) y se usó la técnica de inmunoensayo en fase sólida con el kit de prueba de anticuerpos para brucelosis canina Immunocomb®. De los 123 perros analizados, ninguno dio positivo para los anticuerpos de Brucella abortus mientras que 34 (27.7%) de los perros analizados dieron positivo para los anticuerpos de B. canis. Hubo una asociación significativa (P<0.05) entre infección y género; la infección fue significativamente más alta (P<0.05) en las hembras que en los machos. La prevalencia fue significativamente más alta (P<0.05) en las razas exóticas que en las razas cruzadas y las razas locales. No hubo ninguna relación (P>0.05) entre la infección y los niveles de titulación de anticuerpos en las diferentes categorías de perros. Sin embargo, hubo una relación significativa (P<0.05) entre la presencia de anticuerpos Brucella canis y los perros que andan libremente por doquier. Este estudio provee la primera evidencia serológica de infección con B. canis en perros de los Estados Enugu y Anambra. Esto muestra que la B. canis es endémica en ambos estados, enfatizando la necesidad de hacer más estudios. Las hembras, las razas exóticas y los animales que deambulan libremente se encuentran en el riesgo más alto de infección con Brucella en el área de estudio; por consiguiente, se recomienda enormemente tomar medidas preventivas y de control.

Comparison of immunogenicity between recombinant flagellins C and B of Salmonella abortus equi / 生物工程学报

To evaluate and compare of the immunogenicity differences of flagellins FliC and FljB of Salmonella abortus equi, and lay the experimental foundation for the further utilization of the two recombinant proteins, FliC and FljB recombinant proteins were induced, expressed and purified. The purified FliC and FljB were used to immunize mice separately. The antibody level, titer and subtype of mice serum were detected after immunization. Immune-related receptors and histopathological changes were observed in immunized mice after challenged. The recombinant proteins FliC and FljB were successfully induced and expressed. Proteins of about 52 kDa and 42 kDa were purified. High levels of specific IgG antibodies were induced in mice immunized with these two proteins, the antibody level of FljB-immunized group was higher than that of FliC-immunized group, and IgG1 was the dominant subtype of antibody. The challenge protection rate of the FljB-immunized group was 87.5%, higher than that of FliC immunized group. Bacterial loads and observation pathological of FljB-immunized group were better than that of FliC immunized group, the levels of TCR2, TCR4, MHC-I and TCR induced by FljB-immunized group were higher than those of FliC-immunized group. The of immune response induced by FljB group was better than that of FliC group.

Detección y caracterización molecular de Chlamydia psittaci y Chlamydia abortus en psitácidos mascotas en la provincia de Buenos Aires, Argentina / Detection and molecular characterization of Chlamydia psittaci and Chlamydia abortus in psittacine pet birds in Buenos Aires province, Argentina

In order to determine the presence and genetic diversity of Chlamydia spp. in the north-eastern area of Buenos Aires province, Argentina, conjunctival, oropharyngeal, cloacal swab and tissues were collected from a total of 90 psittacine pet birds of different age and clinical manifestations. Through molecular methods, Chlamydiaceae was detected in 30% (27/90) of the samples, out of which 70.3% (19/27) were positive for Chlamydia psittaci and 14.9% (4/27) for Chlamydia abortus. Nine C. psittaci positive samples were genotyped by ompA gene sequences, 8 clustered within genotype A and 1 within genotype B. A significant association was observed between the presence of Chlamydia spp. and the manifestation of clinical signs compatible with chlamydiosis, as well as with the age of the birds (younger than one year old). This report contributes to the improvement of our understanding of chlamydial agents in our country.

Con el objetivo de determinar la presencia de Chlamydia spp. en psitácidos del área noreste de la provincia de Buenos Aires y conocer su diversidad genética, se recolectaron y analizaron mediante métodos moleculares hisopados conjuntivales, orofaríngeos, cloacales y tejidos de un total de 90 psitácidos de diferentes edades y con diversas manifestaciones clínicas. El 30% (27/90) de las muestras procesadas fueron positivas para Chlamydiaceae; el 70,3% (19/27) de estas resultaron positivas para Chlamydia psittaci y el 14,9% (4/27) para Chlamydia abortus. Nueve muestras positivas para C. psittaci fueron genotipificadas por secuenciación del gen ompA: 8 correspondieron al genotipo Ay una al genotipo B. Se observó una asociación significativa entre la presencia de Chlamydia spp. y la manifestación de signos clínicos compatibles con clamidiosis, como así también con la edad de las aves (menores de un ano). Este informe contribuye a mejorar nuestro conocimiento de los agentes clamidiales en nuestro país.

Bovine brucellosis in slaughterhouses controlled by Federal and Municipal Inspection Services in the state of Maranhão, Brazil / Brucelose bovina em frigoríficos com Serviços de Inspeção Federal e Municipal no estado do Maranhão

The aims of the current study were to investigate the prevalence of bovine brucellosis, to correlate laboratory results of serum reactive samples to the serology of animals presenting serous pouches, and to identify possible risk factors for the development of the disease. In order to do so, 1,265 bovine blood samples were collected from male and female animals bred in different counties in Maranhão state, Brazil, and in other regions of the country. The animals were slaughtered in two slaughterhouses controlled by the Federal Inspection Service (FIS), and in two others controlled by the Municipal Inspection Service (MIS), of regions such as Açailândia and Imperatriz. Samples were tested for specific antibodies by using buffered acidified antigen (BAA) and 2-mercaptoethanol in combination with slow serum agglutination (2-ME + SSA). Additionally, an epidemiological questionnaire was applied to 100 owners, who provided animals to the slaughterhouses, to enable investigating the risk factors for bovine brucellosis. Fifteen serous pouches of animals presenting this lesion were also collected at slaughter time. Thirty-nine out of the analyzed samples were reacted to BAA, whereas 15 reacted to the 2-ME + SSA test; only one male was reagent and it resulted in 1.19% prevalence. One out of the 15 collected serous pouches reacted to the 2-ME + SSA test. The risk factors identified in this study were: the incidence of miscarriages, the sale of animals without previous examination, and the failure in testing the animals before introducing them in herds and before slaughter. It was possible to conclude that the animals slaughtered in the herein evaluated slaughterhouses were infected with Brucella abortus, as well as that this infection prevailed in females.(AU)

Os objetivos do estudo foram determinar a prevalência da brucelose bovina, relacionar resultados laboratoriais de amostras séricas reagentes com a sorologia de animais que apresentavam bursites e, ainda, identificar possíveis fatores de risco para a enfermidade. Para isso, no abate foram coletadas 1.265 amostras de sangue bovino, entre machos e fêmeas, procedentes de diferentes municípios maranhenses e de outras regiões do Brasil, abatidos em dois frigoríficos com Serviço de Inspeção Federal e dois com Serviço de Inspeção Municipal, nas regionais de Açailândia e Imperatriz. As amostras foram testadas para a presença de anticorpos específicos pelos testes do antígeno acidificado tamponado e 2-mercaptoetanol combinado à soroaglutinação lenta (2-ME + SAL). Para realizar o estudo dos fatores de risco, adicionalmente aplicou-se um questionário epidemiológico com 100 proprietários que forneceram animais aos frigoríficos. Foram também coletadas 15 amostras de material de bursite de animais que apresentaram a referida lesão no momento do abate. Do total de amostras analisadas, 39 foram reagentes ao antígeno acidificado tamponado e 15 foram confirmadas no 2-ME + SAL. Apenas um macho foi reagente, o que resultou em prevalência de 1,19%. Das 15 amostras de material de bursites coletadas, uma foi reagente no teste 2-ME + SAL. Os fatores de risco identificados no estudo foram: ocorrência de abortamentos, venda de animais sem exames e não realização de exames que atestem os animais antes da inclusão nos rebanhos para abate. Concluiu-se que a infecção por Brucella abortus em animais abatidos nos frigoríficos avaliados está presente e ocorre com maior frequência em fêmeas.(AU)

Analysis of the risk factors for bovine brucellosis in dairy herds of the Rio Branco microregion, Acre, Brazil / Análise dos fatores de risco para brucelose bovina em rebanhos leiteiros da microrregião de Rio Branco, Acre, Brasil

Bovine brucellosis (BB) is a compulsory notifiable bacterial disease caused by Brucella abortus, which is responsible for severe reproductive disorders in cattle, considerable damages to trade and public health damage. The objectives of this study were to identify the frequency of BB and to analyze the risk factors in the dairy herds of the Rio Branco microregion, in the state of Acre, Brazil. Thus, 527-bovine-serological samples were submitted to the buffered acidified antigen test and to 2-mercaptoethanol. Risk factors were characterized by the administration of epidemiological questionnaires. The logistic regression test was used to evaluate the existence of a significant association between the variables. The frequency of BB in the Rio Branco microregion was 10.6% (56/527). The inappropriate destination of the aborted fetus and placental remnants (odds ratio ­ OR = 13.6), the slaughter of cattle within the property (OR = 3.4) and the age range above 4 years old (OR = 2.9) were the most significant risk factors identified by the logistic regression model. According to the results of this study, it is recommended to intensify BB control and prophylaxis measures in the microregion of Rio Branco, especially those related to sanitary and reproductive management in the livestock breeding.(AU)

A brucelose bovina (BB) é uma bacteriose de notificação obrigatória, causada pela Brucella abortus, responsável por distúrbios reprodutivos graves em bovinos, prejuízos consideráveis ao comércio e agravos em saúde pública. Objetivaram-se com este estudo identificar a frequência de BB e analisar os fatores de risco nos rebanhos leiteiros da microrregião de Rio Branco, Acre. Assim, 527 amostras sorológicas de bovinos foram submetidas ao teste do antígeno acidificado tamponado e ao 2-mercaptoetanol. Os fatores de risco foram caracterizados por aplicação de questionários epidemiológicos. Utilizou-se o teste de regressão logística para avaliar a existência de associação significativa entre as variáveis. A frequência da BB na microrregião de Rio Branco foi de 10,6% (56/527). O destino inadequado do feto abortado e restos placentários (odds ratio ­ OR = 13,6), o abate de bovinos na propriedade (OR = 3,4) e a faixa etária acima de 4 anos (OR = 2,9) foram os fatores de risco mais significativos apontados pelo modelo de regressão logística. Conforme os resultados deste estudo, recomenda-se a intensificação nas medidas de controle e profilaxia da BB na microrregião de Rio Branco, sobretudo relacionadas ao manejo sanitário e reprodutivo nas criações.(AU)

Prevalencia de algunas enfermedades infecciosas en bovinos de resguardos indígenas del cauca, colombia, 2017 / Prevalence of some infectious diseases in cattle of the indigenous resguards in cauca, colombia 2017

RESUMEN La zona del Cauca posee resguardos indígenas que han presentado diferentes problemáticas asociadas a la lucha armada en Colombia, lo que ha limitado que las entidades puedan realizar programas de control de enfermedades bovinas. Por lo anterior, el objetivo de este trabajo fue determinar la prevalencia de algunas enfermedades infecciosas, de carácter reproductivo, en el ganado bovino, de tres resguardos indígenas del Cauca, 2017. Se recolectaron muestras sanguíneas de 30 vacas, para determinar la prevalencia de Neospora caninum, Brucella abortus, diarrea viral bovina (DVB), rinotraqueitis infecciosa bovina (IBR) y Leptospira sp. serovares Hardjo prajitno, bovis y Pomona. Las pruebas, se realizaron por ELISA, rosa de bengala y microaglutinación. Mediante el software R, se efectuó un análisis descriptivo y se determinaron las prevalencias para cada enfermedad, se estimaron las co-infecciones y se realizó un modelo de regresión logística, para determinar el efecto del hato, resguardo, componente racial, grupo etario, número de animales y tamaño del hato, sobre la positividad a las enfermedades. Se encontró una prevalencia del 36%, para N. caninum; 60%, DVB; 30%, IBR; 0%, B. abortus; Leptospira spp. serovares Harjo prajitno, 26%; Hardjo bovis, 12% y Pomona, 10%. De acuerdo con el análisis de riesgo, la seropositividad para DVB e IBR estuvo asociada con el resguardo donde habitaban los animales, en este caso, Tacueyó tuvo más riesgo y estuvo directamente relacionado con el área de los hatos; entre mayor área más animales fueron positivos. En general, se encontró que la situación de la zona es similar a la de otros lugares de Colombia.

ABSTRACT The Cauca region in Colombia has indigenous reservations that have presented different problems associated with the armed conflict, which has limited the entities to carry out bovine disease control programs. Therefore, the aim of this work was to determine the prevalence of some infectious diseases of reproductive traits in cattle of three indigenous resguards in Cauca, 2017. Blood samples of 30 cows were collected to determine the prevalence of Neospora caninum, Brucella abortus, bovine viral diarrhea (BVD), Infectious bovine rhinotracheitis (IBR) and Leptospira sp. serovars Hardjo prajitno, bovis and Pomona. The tests were performed by ELISA, rose bengal and microscopic agglutination. Using R software, a descriptive analysis was carried out and the frequencies for each disease were estimated. In addition, coinfections between diseases were estimated and a logistic regression model was performed to determine the effect of the herd, reservation, racial component, age group, number of animals and size of the farm on the disease positivity. The prevalence was N. caninum 36%, BVD 60%, IBR 30%, B. abortus 0%, Leptospira spp. serovars Hardjo prajitno 26%, Hardjo bovis 12%, y Pomona 10%. According to the risk analysis, the seropositivity for BVD and IBR were associated with the place where the animals lived, in this case Tacueyó, but this was directly related to the size of the herds, since the greater, the area more animals were positive. In general, it was found that the situation in the region is similar to that of other sites in Colombia.

Evaluation of dot-blot test for serological diagnosis of bovine brucellosis

Abstract The objective of this study was to standardize and validate the dot-blot test for the serological diagnosis of bovine brucellosis, compare the results with those found in the 2-mercaptoethanol (2-ME) and complement fixation test (CF), and estimate the relative sensitivity and specificity of the dot-blot compared to these tests. Fifty bovine blood serum samples were used for the test standardization, and 1315 samples were used for evaluation and comparison between the tests; the results were compared using the Kappa indicator. At the end of standardization, it was established as optimal for the antigen obtained from Brucella abortus B19 after passing through a microorganism rupture process, the blood serum samples diluted at 1:100, and the conjugate at 1:30,000. The comparison of the dot-blot results with 2-ME showed Kappa index of 0.9939, sensitivity of 99.48%, and specificity 99.91%, with CF, Kappa index of 0.8226, sensitivity 100% and specificity 95.32%. Using the combination of the test results 2-ME and CF to establish the true condition of the animal, the dot-blot showed relative sensitivity of 100%, and relative specificity of 99.91%. The evaluated test proved to be effective and reliable, besides being easy to handle and interpret the results.

Detección de Chlamydia abortus en pérdidas reproductivas de bovinos en la provincia de La Pampa, Argentina / Detection of Chlamydia abortus in bovine reproductive losses in the province of La Pampa, Argentina

Las pérdidas reproductivas constituyen una causa importante de pérdida económica en el ganado bovino, aunque en más del 50% de los casos la etiología es desconocida. Las especies de la familia Chlamydiaceae han sido asociadas con abortos en bovinos y otras espeChlamydia abortus; cies animales, pero no existen datos al respecto en la República Argentina. El objetivo de este trabajo fue determinar la presencia de Chlamydia spp. y de Chlamydia abortus en pérdidas reproductivas de ganado bovino en La Pampa, Argentina. Se estudiaron 251 muestras provenientes de abortos y mortinatos. Se realizó PCR en tiempo real para la detección de la familia Chlamydiaceae y ArrayTube para la identificación de las especies presentes. Se detectó ADN de la familia Chlamydiaceae en 12 muestras (4,78%); el 83,33% (10/12) correspondió a abortos y el 16,66% (2/12) a mortinatos. El análisis por ArrayTube detectó C. abortus en 5 muestras (1,99% del total, 41,67% de las muestras con detección de Chlamydiaceae). Este trabajo presenta la primera confirmación de la presencia de ADN de diversas especies de Chlamydiaceae (incluida C. abortus) en muestras de pérdidas reproductivas de ganado bovino en Argentina. El valor de prevalencia hallado (4,78%) debe ser tomado como un valor basal, debido al tipo de muestras estudiadas. Se halló material genético de Chlamydiaceae que no coincidió con ninguna de las especies conocidas; esto podría deberse a variantes intraespecie o a especies autóctonas aún no descriptas. Es necesario avanzar en el estudio de la infección por estas bacterias en el ganado bovino de Argentina para conocer su dimensión y analizar su impacto económico y zoonótico, y también para planear medidas de prevención y control.

Reproductive losses linked to an infectious etiology in bovine cattle are a major economic concern worldwide. In Argentina, more than 50% of abortion cases have unknown causes. Species belonging to Chlamydiaceae family are frequent etiologic agents of abortion around the world; however, there is yet no information on their prevalence in Argentina. The objective of this work was to identify Chlamydia spp., and particularly C. abortus in reproductive losses from bovine cattle in La Pampa, Argentina. Real time PCR targeting Chlamydiaceae-specific DNA fragments was performed on 251 samples obtained from bovine abortions and stillborns, and ArrayTube was used for species identification on positive samples. Chlamydiaceae DNA was detected in 12 samples of aborted fetuses (4.78%), 83.33% (10/12) accounting for abortions and 16.66% (2/12) for stillborns. C. abortus was detected by ArrayTube in 5 cases (1.99% of all samples, and 41.67% of Chlamydiaceae positive samples). This study shows the first detection of Chlamydiaceae and C. abortus DNA on reproductive losses of bovine cattle in Argentina, and the described prevalence value (4.78%) should be taken as baseline value due to the type of samples analyzed. Detection of genetic material from Chlamydiaceae not matching any of the studied species could be due to intraspecies variants or local species not yet described. Further research on Chlamydia infections in bovine cattle in Argentina is imperative to describe their range, to analyze their economic and zoonotic implications and to make recommendations about prevention and control measures.

Establishment of detection method for Chlamydophila abortus by using SYBR green real-time PCR and determination of bacterial load in mice / 中国人兽共患病学报

We established a fluorescent quantitative PCR (qPCR)method for the detection of Chlamydophila abortus (C. abortus),and replaced the method of smear staining which has subjective influence on the detection of C.abortus inactivated vaccine titer.According to the conserved sequence of the large cysteine-rich periplasmic protein(envB)of C.abortus,a specific primer was designed and the EnvB-PMD19T positive plasmid was used as the reference standard,optimization condition,sensi-tivity assay,specificity assay,repeatability assay and the bacteria loads of organs from mouse have been done.The results showed that the standard curve established with positive plasmid had a liner response from 1×102copies/μL to 1×106copies/μL with the correlation coefficient of 97%,sensitive for detecting C.abortus with the detection limit of 10 copies/μL,and re-peatable and stable with the coefficients of variation less than 2%.According to the result,the established method can detect the bacteria loads in organ of mouse,which provide a reliable method for evaluation of inactivated C.abortus vaccine.

The significance of the expression of p57, CD34 and Ki-67 in edematous lesions of placental villi / 新乡医学院学报

Objective To observe the expression of p57,CD34 and Ki-67 in complete hydatidiform mole (CHM),partial hydatidiform mole (PHM) and hydropic abortus (HA),so as to investigate the role of the expression in the diagnosis and differential diagnosis of edematous lesions of placental villi.Methods A total of 45 cases of CHM tissue,40 cases of PHM tissue,28 cases of HA tissue and 22 cases of normal pregnancy tissue were collected from January 2003 to December 2013 in Anqiu People's Hospital.The expression of p57,CD34 and Ki-67 in placental villi were detected by immunohistochemistry.Results The positive expression rate of p57 in CHM,PHM,HA and normal pregnancy tissues was 2.22％ (3/45),85.00％ (34/40),89.29％ (25/28) and 95.45 ％ (21/22),respectively;the positive expression rate of p57 in CHM tissues was significantly lower than that in PHM,HA and normal pregnancy tissues (x2 =59.908,57.055,58.238;P ＜ 0.01);there was no significant difference in the positive expression rate of p57 in PHM,HA and normal pregnancy tissues (x2 =0.022,0.681,0.074;P ＞0.05).The expression of CD34 in CHM,PHM,HA and normal pregnancy tissues was 10.27 ± 3.00,11.13 ±2.58,35.57 ± 2.36 and 35.55 ± 2.22 respectively;the expression of CD34 in CHM and PHM tissues was significantly lower than that in HA and normal pregnancy tissues (t =37.89,37.86,39.79,37.40;P ＜ 0.01).There was no significant difference in the expression of CD34 between HA and normal pregnancy tissues (t =1.485,P ＞ 0.05),and there was no significant difference in the expression of CD34 between CHM and PHM tissues (t =1.404,P ＞ 0.05).The positive expression rate of Ki-67 in CHM,PHM,HA and normal pregnancy tissues was 64.44％ (29/45),55.00％ (22/40),14.29％ (4/28) and 9.09％ (2/22) respectively;the positive expression rate of Ki-67 in CHM and PHM tissues was significantly higher than that in HA and normal pregnancy tissues (x2 =18.21,12.61,17.53,11.56;P ＜ 0.01);there was no significant difference in the positive expression rate of Ki-67 between HA and normal pregnant tissues (x2 =0.015,P ＞ 0.05),and there was no significant difference in the positive expression rate of Ki-67 between CHM and PHM tissues (x2 =0.787,P ＞ 0.05).Conclusion p57 helps to identify CHM and other edematous lesions of placental villi,CD34 and Ki-67 help to identify hydatidiform mole and HA.Combined detection of p57,CD34 and Ki-67 is helpful for differential diagnosis of CHM,PHM and HA.

Different invasion efficiencies of Brucella abortus wild-type and mutantsin RAW 264.7 and THP-1 phagocytic cells and HeLa non-phagocytic cells / 대한수의학회지

Brucellosis is one of the common zoonoses caused by Brucella abortus (B. abortus). However, little has been reported on factors affecting invasion of B. abortus into host cells. To investigate cell-type dependent invasion of B. abortus, phagocytic RAW 264.7 and THP-1 cells and non-phagocytic HeLa cells were infected with wild-type and mutant B. abortus, and their invasion efficiencies were compared. The invasion efficiencies of the strains were cell-type dependent. Wild-type B. abortus invasion efficiency was greater in phagocytic cells than in epithelial cells. The results also indicated that there are different factors involved in the invasion of B. abortus into phagocytic cells.

Analysis of protein expression in Brucella abortus mutants with different growth rates by two-dimensional gel electrophoresis and LC-MS/MS peptide analysis

Brucella abortus is a bacterium that causes brucellosis and is the causative agent of worldwide zoonoses. Pathogenesis of the B. abortus infection is complicated, and several researchers have attempted to elucidate the infection mechanism of B. abortus. While several proteins have been revealed as pathogenic factors by previous researchers, the underlying mechanism of B. abortus infection is unresolved. In this study, we identified proteins showing different expression levels in B. abortus mutants with different biological characteristics that were generated by random insertion of a transposon. Five mutants were selected based on biological characteristics, in particular, their growth features. Total proteins of mutant and wild-type B. abortus were purified and subjected to two-dimensional gel electrophoresis. Thirty protein spots of each mutant with expression increases or decreases were selected; those with a change of more than 2-fold were compared with the wild-type. Selected spots underwent liquid chromatography tandem mass spectrometry for peptide analysis. DnaK and ClpB, involved in protein aggregation, increased. SecA and GAPDH, associated with energy metabolism, decreased in some mutants with a growth rate slower than that of the wild-type. Mutants with slower growth showed a decrease in energy metabolism-related proteins, while mutants with faster growth showed an increase in pathogenicity-related proteins.