First genome sequence of Chilean Brucella canis SCL strain provides insights on the epidemiology and virulence factors, explaining differences between geographical origins

BACKGROUND: Brucella canis is the etiological agent of canine brucellosis, a worldwide neglected zoonosis that constitutes one of the major infectious causes of infertility and reproductive failure in dogs. Although genomic information available for this pathogen has increased in recent years, here we report the first genome sequencing of a B. canis strain in Chile, and the differences in virulence genes with other B. canis strains. RESULTS: Genome assembly produced a total length of 3,289,216 bp, N50 of 95,163 and GC% of 57.27, organized in 54 contigs in chromosome I, and 21 contigs in chromosome II. The genome annotation identified a total of 1981 CDS, 3 rRNA and 36 tRNA in chromosome I, and 1113 CDS and 10 tRNA in chromosome II. There is little variation between the different strains and the SCL isolate. Phylogenetic analysis showed that the Chilean SCL strain is closely related to B. canis and B. suis strains. Small differences were found when compared to the Serbian isolate, but all strains shared the same recent common ancestor. Finally, changes in the sequence of some virulence factors showed that the SCL strain is similar to other South American B. canis strains. CONCLUSIONS: This work sequenced and characterized the complete genome of B. canis strain SCL, evidencing the complete presence of all the genes of the virB operon, and minor changes in outer membrane proteins and in the urease operon. Our data suggest that B. canis was introduced from North America and then spread throughout the South American continent.

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.

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.

Prevalence state of canine brucellosis in South Korea during 2015 and 2016 / 대한수의학회지

We investigated the prevalence of canine brucellosis in companion and stray dogs between March 2015 and December 2016 and determined the disease characteristics based on the geographic distribution, sex and age of the dogs in South Korea. We conducted a large-scale survey using serological and bacteriological tests. Samples were collected from 2,394 dogs (1,825 companions and 569 strays). Thirty (1.3%) samples were positive for Brucella canis antibodies including 16 (0.9%) from companion dogs and 14 (2.5%) from stray dogs. Two (1.0%) of the 196 samples cultured from the stray dogs were positive. When compared with male dogs, the female companion and stray dogs had a significantly higher prevalence of brucellosis. Moreover, the prevalence of canine brucellosis was significantly higher in stray dogs older than 6 years and the prevalence of the disease in companion dogs was highest in Incheon (2.1%) and Jeolla (2.1%) provinces. Stray dogs from the Daejeon metropolitan area had the highest prevalence of brucellosis (7.9%). National control measures for canine brucellosis have not previously been implemented. Our findings suggest that appropriate screening tests and control measures are necessary to improve the health of dogs and to protect public health in Korea, particularly with the rapid growth of the companion animal industry.

Epidemiological analysis of brucellosis in Jinzhou City, China, 2001-2015 / 中国人兽共患病学报

We analyzed the epidemiological characteristics of brucellosis in Jinzhou from 2001 to 2015 and provided scientific evidence for the development of prevention and control measures.The descriptive epidemiological analysis was conducted on the incidence data and surveillance result of human brucellosis in Jinzhou during that period.We also conducted correlation analysis of positive rate(sera agglutination titers) and incidence rate of human brucellosis in Jinzhou City.The prevalence of human brucellosis in Jinzhou during 2001-2015 was 0.45/100 000 to 16.53/100 000.The surveillance results showed that the reporting time of human brucellosis case mainly concentrated in March to July,accounting for 66.89％ of the total.In 2003-2015,5 904 blood samples were collected from risk population.Totally,296 were positive for Brucella (5.01％) by sera agglutination titers.From 2007,63 strains of Brucella were isolated from 136 blood samples in which 48 were Brucella melitensis type 3,12 were Brucella melitensis type 1,2 were Brucella melitensis variation type and 1 was Brucella canis.Human brucellosis cases in Jinzhou City is upward trend in recent years and the predominant strain circulated was Brucella melitensis type 3.The situation of dogs carrying Brucella should be given a certain view,and timely elimination of sick dogs should be implemented.

The influence of canine brucellosis on morphofunctional features of epididymal spermatozoa: case report / Influência da brucelose canina nos aspectos morfofuncionais de espermatozoides epididimários: relato de caso

The present work reports a clinical case of a mongrel dog, with serological diagnosis of brucellosis, from which epididymal sperm analysis was performed. Sperm samples were collected from different segments of the epididymis (tail, corpus, and caput). Sperm samples were evaluated for computer-assisted motility analysis (CASA), spermatic morphology, mitochondrial activity and sperm plasmatic membrane and acrosomal integrity. Changes in sperm movement patterns were found (progressive motility, percentage of rapid sperm, percentage of rapid velocity, average pathway, curvilinear velocity, velocity straight line, amplitude of lateral head displacement, straightness and linearity), increase of total morphological defects (51%) and absence of sperm mitochondrial activity (20%) were verified, especially for cauda epididymides. We highlight that such changes can contribute to clinical diagnosis of Brucellosis in dogs and to the use of epididymal sperm in reproductive biotechnologies.(AU)

Relata-se o caso de um cão mestiço, com diagnóstico sorológico para brucelose canina, a partir do qual foram realizadas análises do sêmen epididimário. As amostras espermáticas foram coletadas dos diferentes segmentos epididimários (cabeça, corpo e cauda). Foram realizadas as avaliações de motilidade computadorizada do sêmen (CASA), morfologia espermática, atividade mitocondrial, integridade das membranas plasmática e acrossomal. Houve alteração no padrão de movimentação espermática (motilidade progressiva, espermatozoides rápidos, velocidade média da trajetória, velocidade curvilínea, velocidade linear progressiva, amplitude de deslocamento lateral da cabeça, retilinearidade e linearidade), aumento do total de defeitos morfológicos (51%) e da ausência de atividade mitocondrial espermática (20%) dos espermatozoides, especialmente da cauda do epidídimo. Ressalta-se que tais achados podem contribuir para o diagnóstico clínico da brucelose canina e para a utilização do sêmen epididimário em biotecnologias da reprodução.(AU)

Prevalência e fatores de risco para a leptospirose e brucelose na população canina da Estância Turística de Ibiúna, São Paulo, Brasil / Prevalence and risk factors for leptospirosis and brucellosis in the canine population of the Tourist Resort of Ibiúna, State of São Paulo, Brazil

O objetivo do trabalho foi determinar a prevalência de leptospirose e brucelose por Brucella canis e determinar os fatores de risco associados com a positividade em cães da Estância Turística de Ibiúna, estado de São Paulo, Brasil. Foram examinados 570 animais distribuídos em 4 regiões nos 48 bairros do município, no período de setembro de 2007 a março de 2008. O diagnóstico sorológico da leptospirose foi efetuado com o teste de soroaglutinação microscópica (SAM), e para o diagnóstico de brucelose foi realizado hemocultivo. Dos 570 animais examinados, 187 (32,8%; IC95% 28,9 - 36,8) foram soropositivos para leptospirose, com predomínio de reações para os sorovares Pyrogenes, Autumnalis e Canicola, e 6 (1,05%; IC95% 0,4 - 2,2) foram positivos para brucelose. A variável atividade sexual (OR = 1,73) foi identificada como fator de risco associado à positividade para leptospirose, e o manejo do tipo solto foi considerado fator de risco tanto para leptospirose (OR = 1,96) quanto para brucelose (OR = 10,85). Conclui-se que a leptospirose e a brucelose estão presentes em cães da Estância Turística de Ibiúna, São Paulo, e que a atividade sexual e o acesso irrestrito à rua são condições associadas com a prevalência das infecções.(AU)

The aim of this survey was to determine the prevalence of leptospirosis and brucellosis due to Brucella canis and to determine the risk factors associated with positivity in dogs of the Tourist Resort of Ibiúna, State of São Paulo, Brazil. A total of 570 blood samples were collected from dogs from 4 regions of 48 districts of the county of Ibiúna during the period of September 2007 to March 2008. Serological diagnosis of leptospirosis was performed with the microscopic agglutination test (MAT), and blood culture was used for the diagnosis of brucellosis. Of the 570 dogs used 187 (32.8%; 95%CI 28.9 - 36.8) were seropositive to leptospirosis, with predominance of reactions to serovars Pyrogenes, Autumnalis and Canicola, and 6 (1.05%; 95%CI 0.4 - 2.2) were positive to brucellosis. Variable sexual activity (OR = 1.73) was identified as risk factor associated with the positivity to leptospirosis, and free access to street was considered risk factor for both leptospirosis (OR = 1.96) and brucellosis (OR = 10.85). It is concluded that leptospirosis and brucellosis are present in dogs of the Tourist Resort of Ibiúna, State of São Paulo, and sexual activity and free access to street are conditions associated with the prevalence of infections.(AU)

Identification and genetic characteristics of a Brucella canis strain isolated in Hebei Province / 中华地方病学杂志

Objective A suspected strain of Brucella canis isolated in Hebei Province in 2015 was identified and its genetic characteristics were analyzed.Methods After conventional identification of the bacteria strain,multiplex polymerase chain reaction (M-PCR) and multiple-locus variable-number tandem-repeat analysis (MLVA) were conducted to determine its species and genetic characteristics.Results The strain was identified as Brucella canis by conventional subtyping and M-PCR.Using Panel 1,the strain was genotype 3,using Panel 2A,the strain was genotype 28.It was closely clustered with Brucella canis,and differences of repeated numbers at variablenumber tandem-repeat (VNTR) loci Bruce04,Bruce07,Bruce09 and Bruce16 were also displayed.Conclusions The conventional subtyping and molecular methods can improve the stability and accuracy of the identification.Genetic characteristics of the strain is closely related to that of Brucella canis.

Estudio serológcco de brucelosis canina en dos lbergues del municipio de enviggado, Colombia (2011) / Canine brucelosiis serological study in two shelters of the unicipality of envigado, Colombia (2011)

El objetivo de este estudio fue buscar la presencia de anticuerpos de brucelosis en la población de dos albergues caninos del municipio de Envigado (Antioquia, Colombia) y a tal fin se seleccionaron 54 perros. Se tomaron muestras sanguíneas en 10 machos y 44 hembras que fueron procesadas bajo la técnica de inmunocromatografía para Brucella canis. No hubo diferencia estadística entre albergues en los parámetros ‘sexo’ y ‘raza’ de cada grupo de estudio y tampoco se encontró evidencia serológica de la presencia de brucelosis canina en el grupo de animales muestreados. Los resultados indican que, aunque los animales permanecieron en calidad de abandono durante un período de su vida, su condición feral no era permanente debido a que en el momento del estudio se encontraban en albergues. Las políticas intensivas de recolección se orientan más a la atención de mascotas abandonadas que a aquellos animales ferales (los cuales, con mayor frecuencia, pueden ser portadores de la enfermedad). El tipo de población incluida en el estudio, sumado a la práctica estricta de esterilización inmediata dentro del albergue -que a su vez impide los apareamientos entre individuos-, pueden incidir en la reducción la propagación de la enfermedad en el municipio.

The objective of this study was to search the presence of antibodies of canine brucellosis in the population of two hostels in the municipality of Envigado (Antioquia, Colombia). For this were chosen 54 dogs of two canine shelters. They were taken blood samples in 10 males and 44 females, and they were processed under the immunochromatography technique for Brucella canis. There was no statistical difference between the parameters hostels in sex and race of each study group and no serologic evidence was found of the presence of canine brucellosis in canines sampled group. The results indicate that although the animals remained in quality of abandonment in a period of his life, his feral condition was not permanent since they were in shelters at the time of the study. The intensive policies of collecting from patients, is oriented more to care for pets abandoned, but not for ferine animals (which may be carriers of the disease more frequently). The type of population included in the study, added to the strict practice of immediately sterilization within the shelter -which in turn prevents the mattings between individuals-, both may be reducing the spread of the disease in the municipality.

Brucelosis en hmmbras caninas en montería (Colombia): un problema para la salud públcca / Bruceloss in femmale dogs in monteria (Colombia): a public health problem

La brucelosis canina producida por la cepa rugosa Brucella canis, constituye un riesgo para la salud pública. Debido a que produce infertilidad en ambos sexos; en hembras: abortos tardíos, reabsorciones embrionarias, muertes embrionarias, descargas vaginales de color y olor desagradables y nacimiento de cachorros débiles; en machos: epididimitis, orquitis, atrofia testicular, anormalidades espermáticas; así como puede causar enfermedad en humanos. Mediante un estudio epidemiológico descriptivo de corte transversal y mediante el cálculo para poblaciones, se seleccionaron 62 hembras caninas adultas de diferentes áreas urbanas de Montería (Córdoba) para determinar anticuerpos contra B. canis. Se usó una prueba inmunoensayo cromatográfico de fase sólida comercial. Se elaboró una base de datos en formato Excel en la que se registraron datos anamnésicos y los resultados de laboratorio. Se empleó estadística descriptiva. La seroprevalencia de B. canis fue del 6,45%. El 88,71% de las hembras tuvo historia reproductiva normal y el 11,29% se clasificó como anormal por el reporte de abortos y reabsorciones embrionarias. El 4,84% de las hembras con historial normal fueron seroprevalentes, mientras el historial del 1,61% de las seroprevalentes para B. canis tuvo historial reproductivo anormal. La seroprevalencia determinada en hembras caninas hace pensar que se debe tener presente esta enfermedad para su correcto diagnóstico, y la presencia de serorreactores es una evidencia y un riesgo epidemiológico para la salud pública.

Canine brucellosis caused by the rough strain of Brucella canis, constitutes a risk to public health because it produces infertility in both sexes; in female it causes late-term abortions, embryo resorption, embryonic death, vaginal discharges with unpleasant colors and odors, weak newborn puppies; in males it causes epididymitis, orchitis, testicular atrophy and sperm abnormalities; it can also cause illness in humans. Through a descriptive cross section epidemiological study and by calculation for populations, 62 adult canine females of different urban areas of Montería (Córdoba) were selected to determine antibodies against B. canis, using a commercial chromatographic immunoassay test in solid phase. An Excel database format was developed in order to register anamnestic data and laboratory results. In addition, descriptive statistics was employed in this study. The results of this study show that B. canis seroprevalence was 6.45%; 88.71% of the female's reproductive histories was normal; and 11.29% were classified as abnormal by the report of abortions and embryonic resorption; 4.84% of the females with normal reproductive history were seroprevalent while 1.61% of the seroprevalent for B. canis had an abnormal reproductive history. The seroprevalence determined in females dogs suggests that this disease must be considered for its correct diagnosis and the presence of seroreactants is an evidence and an epidemiological risk for public health.

Infección por Brucella canis en humanos: propuesta de un modelo teórico de infección a través de la ruta oral / Brucella canis infection in humans: Proposal of a theoretical model of infection through the oral route

La infección por Brucella canis en los humanos se ha reconocido recientemente como una zoonosis, pero frecuentemente es sub reportada debido a que los síntomas pueden confundirse con los de un resfriado común u otras infecciones causadas por otros patógenos. Los caninos son los hospederos primarios de Brucella canis ; el incremento en la tendencia de tener perros como mascotas podría también aumentar la posibilidad de transmisión de la infección a los humanos por el estrecho contacto entre la mascota infectada y su propietario. En Colombia, hay reportes de aislamientos de B. canis de caninos de criaderos y de un humano en contacto con perros infectados, al igual que reportes de caninos seropositivos a la infección. Sin embargo, no hay mucha información disponible sobre los mecanismos de interacción hospedero-patógeno que conduzcan al establecimiento de la infección por Brucella canis en perros y en humanos esta información es todavía menor. En esta revisión se propone un modelo para la infección humana con Brucella canis a través de la ruta oral utilizando la información disponible para otras especies de Brucella que infectan al humano, incluyendo B. abortu s y B. melitensis , que difieren de B. canis en la composición estructural de su lipopolisacárido. También se hipotetiza el mecanismo de infección celular que es usado por B. canis para invadir y establecer la infección en células no fagocíticas y fagocíticas.

Brucella canis infection in humans has recently been recognized as a zoonosis, but it is frequently under reported because the flu-like symptoms are often confused with the presence of other disease-causing pathogens. Dogs are the primary hosts for Brucella canis ; the increasing trend to adopt dogs as pets also enhances the likelihood of transmission of Brucella canis infection through contact between infected dogs and owners. In Colombia, there are reports of isolates of B. canis from kennel dogs and also from one human being along with seropositive results from dogs and humans. However, the mechanism of hostpathogen interactions leading to the infection of Brucella canis in dogs is still unknown and even less is known about human infections. This review proposes a model for human infection with Brucella canis through the oral route. We use the information available for other human-infecting Brucella species, including B. abortu s and B. melitensis, which differ from B. canis in the structural composition of the lipopolysaccharide molecule. The mechanism of cellular infection used by B. canis to invade and establish infection in nonphagocytic and phagocytic cells is also hypothesized.

Detección de Brucella canis por método de inmunocromatografía en perros vagos capturados en la ciudad de Temuco, Chile, 2011 / Detection of Brucella canis by immunochromatography method in vague dogs captured in Temuco city, Chile, 2011

Background: Brucella canis is responsible for brucellosis in dogs, causing reproductive disorders and is considered a zoonoses, as described in several countries. The epidemiological data are scarce in our country. Aim: To determine the prevalence of Brucella canis in vague dogs in Temuco city and housed in the Temuco Kennel. Methods: Quantitative and cross-section study. We used 400 samples of dogs of both sexes, different ages and mainly mixed race, which were tested by immunochromatography. Results: Antibodies were detected in 4 samples Brucella canis which represented 1% of the population studied, 2 females (0.5%) and 2 males (0.5%). Discussion: We conclude that dogs are infected by B. canis in a low range but remains a risk condition to the health of the human population if not maintained adequate sanitary control of pets, like vague dogs.

Introducción: Brucella canis es responsable de la brucelosis en perros, provocándoles trastornos reproductivos y es considerada una zoonosis, ya descrita en varios países. Los datos epidemiológicos en nuestro medio son exiguos. Objetivo: Determinar la prevalencia de B. canis en perros vagos capturados en la ciudad de Temuco y albergados en el Canil Temuco. Materialy Métodos: Estudio de tipo cuantitativo y de corte transversal. Se utilizaron 400 muestras de perros de ambos sexos, diferentes edades y principalmente mestizos, procesadas mediante la prueba de inmunocromatografía. Resultados: Se detectaron anticuerpos anti-B. canis en 4 muestras lo cual representó 1% de la población estudiada, 2 hembras (0,5%) y 2 machos (0,5%). Conclusión: El hallazgo de perros serológicamente positivos a B. canis, es baja pero no deja de ser un indicador del riesgo en el que se encuentra la salud de la población humana si no se mantiene un adecuado control sanitario de las mascotas, como ocurre con los perros vagos.

Toxoplasmosis, leptospirosis and brucellosis in stray dogs housed at the shelter in Umuarama municipality, Paraná, Brazil

Background : Leptospirosis, toxoplasmosis and brucellosis are diseases with worldwide distribution. Among stray dogs, these zoonoses are facilitated by direct contact with other animal species, by the habit of scavenging garbage and hunting in search of food, drinking standing water, smelling other animals' urine, licking female genitalia and the sexual act itself. The objective of this study was to detect antibodies anti-Toxoplasma gondii, anti-Leptospira spp., anti-Brucella canis and anti-Brucella abortus in stray dogs housed in shelters at Umuarama city, Paraná, Brazil. In order to detect toxoplasmosis, indirect immunofluorescence assay (IFA) was performed, agglutination microscopic (MAT) test for leptospirosis and agar gel immunodiffusion (AGID) and buffered acidified antigen (BAA) tests for brucellosis. Results : Of the 175 serum samples analyzed, 70.85% were considered positive for toxoplasmosis by IFA, 20% by MAT for leptospirosis and 2.85% by AGID for Brucella canis. Conclusions : The serological results of this study showed that stray dogs housed at the private shelter are potential carriers of these three different zoonoses and contribute to the spread and maintenance of these etiologic agents in the urban area of Umuarama (PR), Brazil.(AU)

Toxoplasmosis, leptospirosis and brucellosis in stray dogs housed at the shelter in Umuarama municipality, Paraná, Brazil

Background : Leptospirosis, toxoplasmosis and brucellosis are diseases with worldwide distribution. Among stray dogs, these zoonoses are facilitated by direct contact with other animal species, by the habit of scavenging garbage and hunting in search of food, drinking standing water, smelling other animals' urine, licking female genitalia and the sexual act itself. The objective of this study was to detect antibodies anti-Toxoplasma gondii, anti-Leptospira spp., anti-Brucella canis and anti-Brucella abortus in stray dogs housed in shelters at Umuarama city, Paraná, Brazil. In order to detect toxoplasmosis, indirect immunofluorescence assay (IFA) was performed, agglutination microscopic (MAT) test for leptospirosis and agar gel immunodiffusion (AGID) and buffered acidified antigen (BAA) tests for brucellosis. Results : Of the 175 serum samples analyzed, 70.85% were considered positive for toxoplasmosis by IFA, 20% by MAT for leptospirosis and 2.85% by AGID for Brucella canis. Conclusions : The serological results of this study showed that stray dogs housed at the private shelter are potential carriers of these three different zoonoses and contribute to the spread and maintenance of these etiologic agents in the urban area of Umuarama (PR), Brazil.

Seroprevalencia y factores de riesgo para brucelosis canina en perros domésticos de once comunas de la ciudad de Medellín-Colombia / Canine brucellosiss: Seroprevalence and risk factors in pets from eleven neighbourhoods in Medellin, Colombia

Objetivo: Se estableció la seroprevalencia de brucelosis canina en perros domésticos de 11 comunas de la ciudad de Medellín y se realizó una encuesta de factores de riesgo a los propietarios. Materiales y Métodos: Fueron analizados por inmunoensayo cromatográfico 441 muestras de suero de caninos cuyos dueños aceptaron responder a una encuesta epidemiológica para explorar factores de riesgo de transmisión de brucelosis canina a la población humana. Resultados: La seroprevalencia de brucelosis fue 2,76 % (IC: 1,11 %- 4,42 %) siendo mayor para las comunas Buenos Aires (6,9 %) y Villa Hermosa (5,7 %). La mayor seroprevalencia fue en machos (4,6 %), caninos criollos (4,8 %), menores de un año (3,7 %), caninos de viviendas con fuentes de agua cercanas (4,5 %), y en aquellos que han permanecido con el dueño por más de 5 meses (3,1 %). La raza presentó asociación con la presencia de anticuerpos (p < 0,05). La seroprevalencia aumentó a 7,5 % cuando la vivienda era compartida con otros animales domésticos. Se encontró mayor seroprevalencia entre los caninos que habitaban en viviendas sin suministro de agua potable (6,7 %) y sin conexión de acueducto y alcantarillado (7,7 %). Para ninguna de estas variables se encontró asociación con la presencia de anticuerpos. Conclusión: El riesgo de transmisión a los humanos de esta zoonosis emergente en circunstancias diferentes a las de índole ocupacional, como son los ambientes domésticos, puede considerarse en aumento si persisten o se incrementan los factores de riesgo que se exploraron en este estudio.

Objective: Determining Brucella canis prevalence in domestic dogs (pets) from 11 districts in Medellín. A survey of risk factors was also carried out. Materials and Methods: Immunoassay was used for analysing441 dog serum samples and several risk factors regarding their owners and some related to the immediate environment were established. Results: Brucellosis prevalence was 2.76 % (1.11-4.42 95 %CI), being highestin the Buenos Aires (6.9 %) and Villa Hermosa districts (5.7%). Seroprevalence was higher in male dogs (4.6 %),mongrels (4.8 %),dogs less than one year old (3.7 %), in homes having nearby water sources (4.5 %) andindogs living with their owners for more than five months (3.1 %).The dogs’ breed was associated with antibody presence (p<0.05). Seroprevalence became increased to 7.5 % when the home was shared with other pets. Higher prevalence was found when the dogs lived in homes without drinking water (6.7 %) and homes lacking a fixed water supply or sewerage connection (7.7 %). No association was found for any of the aforementioned variables with the presence of antibodies. Conclusion: Apart from cases involving occupational risk, the risk ofBrucella canistransmission to humans in domestic settings may increase if the aforementioned risk factors continue increasing in urban areas, such as those explored in this study.

Diagnosis of canine brucellosis using recombinant ribosomal protein L7/L12 / 대한수의학회지

Brucella (B.) canis is mainly transmitted by direct or indirect contact with aborted fetuses and placenta. It's also known to be able to infect human, which likely results in providing veterinarians and companion animal owners for infectious risk. To develop diagnostic ELISA, we cloned and expressed rp1L gene of B. canis, which encodes the ribosomal protein L7/L12. Using this purified recombinant protein, indirect-ELISA (iELISA) was evaluated using 78 positive and 44 negative sera. The sensitivity and the specificity of iELISA were 94% and 89%, respectively. The results indicated that indirect-ELISA using recombinant ribosomal protein L7/L12 may be useful for diagnosis of canine brucellosis.

Seroprevalencia de Brucella canis en perroscallejeros del Centro de Bienestar Animal La Perla,Medellín (Colombia), 2008 / Seroprevalência da Brucelose canis em cachorros de rua do Centro de Bem-estarAnimal La Perla, Medellín (Colômbia), 2008 / Brucella canis seroprevalence in stray dogs from “Centro de Bienestar Animal La Perla, Medellín (Colombia), 2008

Brucella canis es el principal agente causal de enfermedad reproductiva en caninos. Por su potencialzoonótico y su distribución mundial, esta infección tiene una gran importancia. En Colombia, la bacteriase ha aislado de mascotas caninas, de perros de criadero, y de humanos. Este estudio se realizó con elobjetivo de determinar la frecuencia de esta infección en perros callejeros, alojados en el Centro deBienestar Animal “La Perla” (Medellín, Colombia). En junio de 2008 se muestrearon 221 caninos y serealizó prueba de aglutinación rápida en placa. Se encontró una seroprevalencia del 6.78%, y no hubodiferencia por grupos de edad o sexo. Los resultados alertan sobre la población de perros callejeros comoun factor de riesgo para la transmisión de la enfermedad.

Brucella canis is the main causal agent of reproductive illness in dogs. The disease has a zoonoticpotential and is distributed worldwide, hence his great importance. In Colombia, the bacterium has been isolated from pet dogs, kennel dogs, and also from human beings. The objective of this study was to takea look at the frequency of B. canis infection in street dogs sheltered in the Center for Animal Welfare “LaPerla” (Medellín, Colombia). In June 2008, 221 dogs were sampled and their sera were tested by the rapidslide agglutination test. Seroprevalence was 6.78%,and no difference was found between different age orsex groups. The results alert about the street dogs population as a risk factor for the transmission of theBrucella canis.

Brucella canis é o principal agente causador da doença reprodutiva em cães. Por seu potencial zoonóticoe sua distribuição global, esta infecção é de grande importância. Na Colômbia, a bactéria foi isolada decães, canil, e os seres humanos. Este estudo foi conduzido com o objetivo de fazer uma abordagem àfreqüência desta infecção em cães de rua, alojados no Centro de Bem-Estar Animal "Pearl" (Medellín,Colômbia). Em junho de 2008, foram amostrados 221 cães e foi feito o teste rápido de aglutinação. Foiencontrada uma soroprevalência de 6.78%, e não houve diferença de idade e do sexo. Os resultados alertampara a população de cães de rua como um fator de risco para transmissão da doença.

Brucelosis canina en perros de la ciudad de Buenos Aires / Canine brucellosis in dogs in the city of Buenos Aires

La brucelosis canina, causada por Brucella canis, provoca epididimitis, atrofia testicular y esterilidad en los perros, mientras que en las hembras el síntoma principal es el aborto. La transmisión al hombre puede ser por contacto con el semen, orina, y/o fetos abortados de animales infectados. El presente estudio de tipo observacional de corte transversal, se realizó en caninos de barrios y asentamientos con alto índice de necesidades básicas insatisfechas (NBI) en 8 áreas de la ciudad de Buenos Aires. Se estudiaron 219 perros, 184 hembras y 35 machos, que fueron negativos a la prueba de aglutinación con antígeno tamponado (BPAT), que descartó la infección con especies lisas del género Brucella. Sedetectaron anticuerpos anti-B. canis en 16 perros (7.3%), 9 hembras y 7 machos, este último dato es relevante ya que la orina de los machos es considerada uno de los medios de diseminación de la infección. Aunque sólo se pudieron tomar hemocultivos a 175 animales, en 3 (2 hembras y un macho) se aislaron B. canis. Sólo 3 de los dueños de los perros positivos accedieron al diagnóstico serológico y dos resultaron positivos. Destacamos que la prueba de inmunodifusión en gel de agar (IGDA) ha demostrado ser poco sensible, detectó sólo uno de los 16 casos positivos y ninguno de los tres confirmados por aislamiento. Concluimos que en las áreas estudiadas el hallazgo de perros serológicamente positivos y el aislamiento de B. canis en 3 casos, son indicadores del riesgo en el que se encuentra la salud de la población expuesta.

Canine brucellosis, caused by Brucella canis, provokes epidydimitis, testicular atrophy and sterility in male dogs, while in females the major symptom is miscarriage. Transmission to humans may be through contact with semen, urine and/or aborted fetuses of infected animals. Our study, observational and cross-sectional, focused on dogs in lower class neighborhoods and slums with a high rate of unmet basic needs (UBN) in 8 areas of the city of Buenos Aires. We studied 219 dogs: 184 females and 35 males, that tested negative to the buffered plate antigen test (BPAT), which ruled out infection with smooth species of Brucella. We detected anti-B. canis antibodies in 16 dogs (7.3%): 9 females and 7 males, relevant data since the urine of males is considered one of the vectors for the spread of the infection. Although we could run blood cultures on only 175 animals, we isolated B. canis in 3 (2 females and 1 male). Only 3 of the owners of dogs that tested positive consented to a serological diagnosis and two of them were positive. We highlight that the agar gel immunodiffusion test (IGID) proved to have low sensitivity, having detected only one of the 16 positive cases and none of the three confirmed by isolation. We conclude that in the areas studied, the detection of serologically positive dogs and the isolation of B. canis in 3 cases are indicators of the health hazard for the population exposed to it.

Pesquisa de anticorpos anti-Brucella canis e anti-Brucella abortus em cães errantes da cidade de SãoJoão da Boa Vista, Estado de São Paulo, Brasil(2002-2003) / Research of antibodies anti-Brucella canis and anti-Brucella abortus inwanderring dogs in São João da Boa Vista, State of São Paulo, Brazil(2002-2003)

A brucelose canina é uma importante zoonose que tem sido investigada freqüentemente em cães como medida sanitária de controle da saúde pública. Foi conduzido um estudo sorológico para investigar a freqüência da brucelose canina por Brucella canis e por Brucella abortus, em 500 cães errantes na cidade de São João da Boa Vista/SP - Brasil, utilizando as técnicas de imunodifusão em gel de ágar (antígeno de parede celular de B. ovis) e imunoaglutinação em placa com antígeno acidificado tamponado. Este estudo mostrou baixa frequência de cães infectados por B. canis 4/500 (0,8%) e ausência de soros positivos para B. abortus

Dogs brucellosis may cause reprodutive problems and there are a risk factors for human health. The prevalence of brucellosis due to Brucella canis and Brucella abortus was investigated in dogs living in urban areas of São João da Boa Vista/SP - Brasil. For this purpose, 500 blood sample were collected and serum analysed by agar gel immuno diffusion test and Rose-Bengal test. The frequency of brucellosis due to B. canis was 4/500 (0,8%) and negative for B. abortus.

Brucella spp. isolation from dogs from commercial breeding kennels in São Paulo state, Brazil

Dogs from 12 commercial breeding kennels were submitted to clinical investigation and laboratorial tests for diagnosis of Brucella spp. infection. The sampling was carried out between April 2000 and February 2002 and the laboratorial tests employed were agar gel immunediffusion test (AGID) and blood culture. From 171 dogs examinated, 39 (22.8%) showed at least one clinical sign compatible with brucellosis, 58 (33.91%) were AGID positive and 24 (14.03%) were positive by blood culture. Gram negative bacterial cells with a biochemical pattern compatible with that of bacteria belonging to genus Brucella were isolated from blood specimens of 24 animals. According to Kappa index and McNemar test, the association between AGID and blood culture (k=0.360 with 95% of confidence interval; X²=25.93, p=0.000), between AGID and clinical test (k=0.248 with 95% of confidence interval; X²=6.11, p=0.013), and between blood culture and clinical examination (k=0.442 with 95% of confidence interval; X²=6.76, p=0.009) were not statistically significant. Qui-Square test indicated no association of sex and the results of clinical examination (X²=1.35 and p=0.2447), AGID (X²=1.58 and p=0.2086) or bacterial isolation (X²=1.48 and p=0.2230). Within 12 kennels, seven had at least one dog positive by blood culture and nine had at least one animal positive by AGID. The association of epidemiological data with direct and indirect methods of diagnosis is necessary to perform a definitive diagnosis of Brucella infection in dogs, as positive results by AGID can be consequence of non-specific reactions and must be confirmed by blood culture. Negative results by AGID must also be confirmed using direct methods of diagnosis or repeating the serologic test after 30 days, because of the low sensitivity of this test.

Cães provenientes de 12 canis comerciais do estado de São Paulo foram submetidos à investigação clínica e a provas laboratoriais para o diagnóstico de infecção por Brucella spp. A colheita de amostras foi realizada entre os meses de abril de 2000 e fevereiro de 2002 e os exames laboratoriais empregados foram a imunodifusão em gel de ágar (IDGA) e a hemocultura. De 171 cães examinados, 39 (22,80 %) apresentaram pelo menos um sinal clínico compatível com brucelose, 58 (33,91%) foram positivos pela IDGA e 24 (14,03%) pela hemocultura. Bactérias Gram negativas com perfil bioquímico compatível com o gênero Brucella foram isoladas das 24 amostras de sangue positivas pelo isolamento bacteriano. De acordo com o coeficiente Kappa e o teste de McNemar, não foi observada concordância entre os resultados obtidos na hemocultura e IDGA (k=0,360 com intervalo de confiança de 95%; X²=25,93, p=0,000), entre resultados da IDGA e do exame clínico (k=0,248 com intervalo de confiança de 95%; X²=6,11, p=0,013) e entre os resultados da hemocultura e do exame clínico (k=0,442 com intervalo de confiança de 95%; X²=6,76, p=0,009). A associação dos resultados obtidos pelos exames clínicos e laboratoriais com o sexo dos animais não foi estatisticamente significante (Qui-Quadrado), sendo observado X²=1,35 e p=0,2447 para o exame clínico, X²=1,58 e p=0,2086 para IDGA e X²=1,48 e p=0,2230 para hemocultura. Dos 12 canis examinados, sete apresentaram pelo menos um animal positivo pela hemocultura e nove pelo menos um animal positivo pela imunodifusão. A associação de dados epidemiológicos com testes laboratoriais diretos e indiretos deve ser enfatizada para o diagnóstico definitivo da brucelose canina. Resultados positivos pela imunodifusão em gel de ágar podem ser conseqüência de reações inespecíficas e devem ser confirmados pela hemocultura. Os resultados negativos obtidos pela imunodifusão também devem ser confirmados utilizando-se métodos diretos de diagnósticos ou repetindo-se o teste sorológico com 30 dias de intervalo, devido à baixa sensibilidade desse teste diagnóstico.