RESUMO
Animal hoarding disorder (AHD) is classified as a psychiatric obsessive-compulsive condition characterized by animal accumulation and often accompanied by unsanitary conditions and animal cruelty. Although AHD may increase pathogen transmission and spread, particularly for zoonotic diseases, human and dog exposure in such cases has yet to be fully established. Accordingly, this study aimed to assess Brucella canis in 19 individuals with AHD (11 households) and their 264 dogs (21 households) in Curitiba, the eighth largest city in Brazil, with approximately 1.8 million habitants. Anti-B. canis antibodies were detected by the 2-mercaptoethanol microplate agglutination test (2ME-MAT) and by a commercial lateral flow immunoassay (LFIA), while molecular detection of previously positive seropositive samples was performed by conventional PCR. Although all the human samples were 2ME-MAT negative, 12/264 (4.5%, 95% Confidence Interval: 2.0-7.0%) dog samples were 2ME-MAT and LFIA positive, with 2ME-MAT titers ranging from 20 to 640. At least one dog in 4/21 (19.0%, 95% CI: 2.0-46.0%) households was seropositive. Despite the absence of seropositivity in individuals with AHD and the comparatively low seroprevalence in dogs, B. canis circulation and outbreaks should be considered in such human populations due to the high burden and recurrent character of B. canis exposure in high-density dog populations and the constant introduction of susceptible animals.
Assuntos
Brucella canis , Brucelose , Doenças do Cão , Transtorno de Acumulação , Animais , Cães , Humanos , Brucella canis/genética , Brucelose/diagnóstico , Brucelose/epidemiologia , Brucelose/veterinária , Doenças do Cão/diagnóstico , Doenças do Cão/epidemiologia , Saúde Única , Estudos SoroepidemiológicosRESUMO
Background: Although Brucella abortus, Brucella suis, and Brucella canis may infect humans and dogs worldwide, no study to date has assessed and compared owners and their dogs between island and mainland seashore areas. Materials and Methods: Accordingly, the study herein has applied serological tests, including Microplate Agglutination Test with 2-Mercaptoethanol, immunochromatographic assay, and Rose Bengal Test, and a Brucella genus-specific PCR assay to 195 owners and their 148 dogs living on 1 mainland seashore area and three nearby oceanic islands of southern Brazil. Results: No seropositivity to B. abortus and B. suis was detected in owner or dog sera. Anti-B. canis seropositivity was observed in 3/148 (2.0%) dogs, but no owner sample was seropositive to B. canis. In addition, all blood samples from both owners and dogs were negative on Brucella genus-specific PCR assay. Conclusions: The seropositive dogs were not related and lived on the seashore mainland area of Guaraqueçaba city. The absence of seropositivity on the islands and the low seropositivity on the seashore mainland could be attributed to geographic isolation, and suggest the low impact of the disease in the region. Despite being a zoonotic disease, brucellosis by B. canis is not included in the National Program for Control and Eradication of Brucellosis, and its diagnosis and notification are not mandatory. The presence of seropositive dogs highlights the risk to human health and the importance of epidemiological surveillance actions in the region, as well as the need for the implantation of preventive measures to avoid the transmission of the pathogen.
Assuntos
Brucella canis , Brucelose , Doenças do Cão , Humanos , Cães , Animais , Brasil/epidemiologia , Doenças do Cão/epidemiologia , Brucelose/epidemiologia , Brucelose/veterinária , Brucelose/diagnóstico , Brucella canis/genética , Brucella abortusRESUMO
Brucella canis is responsible for canine brucellosis, a neglected zoonotic disease. The omp25 gene has been described as an important marker for Brucella intra-species differentiation, in addition to the ability to interact with the host immune system. Therefore, this study investigated the omp25 sequence from B. canis strains associated to a phylogenetic characterization and the unveiling of the molecular structure. In vitro analyses comprised DNA extraction, PCR, and sequencing of omp25 from 19 B. canis strains. Moreover, in silico analyses were performed at nucleotide level for phylogenetic characterization and evolutionary history of B. canis omp25 gene; and in amino acid level including modeling, dynamics, and epitope prediction of B. canis Omp25 protein. Here, we identified a new mutation, L109P, which diverges the worldwide omp25 sequences in two large branches. Interestingly, this mutation appears to have epidemiology importance, based on a geographical distribution of B. canis strains. Structural and molecular dynamics analyses of Omp25 revealed that Omp25L109P does not sustain its native ß-barrel. Likewise, the conformation of B-cell epitope on the mutated region was changed in Omp25L109P protein. Even without an evolutive marker, the new identified mutation appears to affect the basic function of B. canis Omp25 protein, which could indicate virulence adaptation for some B. canis strains in a context of geographical disposition.
Assuntos
Proteínas de Bactérias , Brucella canis , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Brucella canis/classificação , Brucella canis/genética , Brucella canis/fisiologia , Evolução Molecular , Genes Bacterianos , Modelos Moleculares , Mutação , Filogenia , Reação em Cadeia da Polimerase , Conformação Proteica , Análise de Sequência de DNARESUMO
La prevalencia de la brucelosis canina por B. canis, está aumentando a nivel mundial, al igual que los reportes en personas. La ausencia de vacunas y de programas de control, permiten predecir una rápida expansión de la enfermedad, con el consecuente impacto en la reproducción canina y en la Salud Pública. Por lo anterior, es urgente analizar candidatos vacunales que consideren las características genómicas de las cepas circulantes a nivel mundial, toda vez que se ha comprobado variabilidad genética, particularmente en genes asociados a factores de virulencia tales como proteínas de membrana externa y sistema de secreción tipo IV. La presencia de un conglomerado genético de cepas Latinoamericanas de B. canis debe ser considerado en estudios futuros de tal manera de generar productos vacunales sin limitaciones geográficas.(AU)
A prevalência da brucelose canina por B. canis está aumentando em todo o mundo, assim como os relatos em humanos. A falta de vacinas e programas de controle prevêem uma rápida disseminação da doença, com conseqüente impacto sobre a reprodução canina e a saúde pública. Portanto, é urgente analisar os candidatos a vacina que consideram as características genômicas das cepas que circulam pelo mundo, uma vez que a variabilidade genética foi comprovada, particularmente nos genes associados a fatores de virulência, como as proteínas da membrana externa e o sistema de secreção tipo IV. A presença de um grupo genético de linhagens latino-americanas de B. canis deve ser considerada em estudos futuros a fim de gerar produtos vacinais sem limitações geográficas.(AU)
Assuntos
Variação Genética , Brucelose/prevenção & controle , Infertilidade/veterinária , Reprodução/genética , Brucella canis/genética , Desenvolvimento de Vacinas/métodosRESUMO
Brucellosis is a prevalent disease in Costa Rica (CR), with an increasing number of human infections. Close to half of homes in CR have one or more dogs, corresponding to â¼1.4 million canines, most of them in the Central Valley within or near the cities of San José, Heredia, and Alajuela. From 302 dog sera collected from this region, 19 were positive for Brucella canis antigens, and five had antibodies against smooth lipopolysaccharide, suggesting infections by both B. canis and other Brucella species. B. canis strains were isolated in the Central Valley from 26 kennel dogs and three pet dogs, all displaying clinical signs of canine brucellosis. We detected three recent introductions of different B. canis strains in kennels: two traced from Mexico and one from Panama. Multiple locus-variable number tandem repeats (MLVA-16) and whole-genome sequencing (WGSA) analyses showed that B. canis CR strains comprise three main lineages. The tree topologies obtained by WGSA and MLVA-16 just partially agreed, indicating that the latter analysis is not suitable for phylogenetic studies. The fatty acid methyl ester analysis resolved five different B. canis groups, showing less resolution power than the MLVA-16 and WGSA. Lactobacillic acid was absent in linages I and II but present in linage III, supporting the recent introductions of B. canis strains from Mexico. B. canis displaying putative functional cyclopropane synthase for the synthesis of lactobacillic acid are phylogenetically intertwined with B. canis with non-functional protein, indicating that mutations have occurred independently in the various lineages.
Assuntos
Brucella canis/genética , Brucelose/epidemiologia , Brucelose/veterinária , Surtos de Doenças/veterinária , Doenças do Cão/microbiologia , Filogenia , Animais , Brucella canis/classificação , Brucella canis/patogenicidade , Costa Rica/epidemiologia , Doenças do Cão/epidemiologia , Cães , Evolução Molecular , Feminino , Variação Genética , Genoma Bacteriano , Genótipo , Espécies Introduzidas , Masculino , México , Panamá , Animais de Estimação/microbiologia , Sequenciamento Completo do GenomaRESUMO
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.
Assuntos
Animais , Cães , Brucelose/epidemiologia , Brucella canis/genética , Brucella canis/patogenicidade , Urease/genética , Brucelose/transmissão , Zoonoses , Chile , GenomaRESUMO
The aim of this study was to estimate the diversity and prevalence of both groups of Brucella canis 1 and 2 with and without deletion respectively in different areas of Argentina. A total of 104 bacterial cultures were typed as B. canis strains using the classical biotyping method. Two PCR assays were performed to confirm that all isolates were B. canis and not Brucella suis. The differentiation between groups 1 and 2 was achieved using another PCR assay and the diversity of B. canis isolates was assessed with four MLVA_16 markers. All strains belonged to Group 2. Bruce 09 marker (MLVA_16 assay) showed the greatest diversity. Only Group 2 of B. canis was identified among the strains evaluated. The markers chosen from the MLVA_16 allowed us to detect genetic diversity among the strains of B. canis studied.
Assuntos
Brucella canis , Brucella suis , Brucelose , Argentina/epidemiologia , Brucella canis/genética , Brucelose/epidemiologia , Brucelose/veterinária , Humanos , Reação em Cadeia da PolimeraseRESUMO
Brucellosis is one of the most common bacterial zoonoses worldwide affecting not only livestock and wildlife but also pets. Canine brucellosis is characterized by reproductive failure in dogs. Human Brucella canis infections are rarely reported but probably underestimated due to insufficient diagnostic surveillance. To improve diagnostics, we investigated dogs in a breeding kennel that showed clinical manifestations of brucellosis and revealed positive blood cultures. As an alternative to the time-consuming and hazardous classical identification procedures, a newly developed species-specific intact-cell matrix-assisted laser desorption/ionization-time of flight mass spectrometry analysis was applied, which allowed for rapid identification of B. canis and differentiation from closely related B. suis biovar 1. High-throughput sequencing and comparative genomics using single nucleotide polymorphism analysis clustered our isolates together with canine and human strains from various Central and South American countries in a distinct sub-lineage. Hence, molecular epidemiology clearly defined the outbreak cluster and demonstrated the endemic situation in South America. Our study illustrates that MALDI-TOF MS analysis using a validated in-house reference database facilitates rapid B. canis identification at species level. Additional whole genome sequencing provides more detailed outbreak information and leads to a deeper understanding of the epidemiology of canine brucellosis.
Assuntos
Brucella canis , Brucelose , Surtos de Doenças , Doenças do Cão , Genoma Bacteriano , Polimorfismo de Nucleotídeo Único , Animais , Brucella canis/genética , Brucella canis/metabolismo , Brucelose/sangue , Brucelose/epidemiologia , Brucelose/genética , Brucelose/veterinária , Doenças do Cão/sangue , Doenças do Cão/epidemiologia , Doenças do Cão/genética , Cães , Genômica , Sequenciamento de Nucleotídeos em Larga Escala , América do Sul/epidemiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Canine brucellosis is an infectious disease that produces reproductive disease in both males and females. Although Brucella canis is more common, the infection by Brucella abortus is more frequent in dogs sharing habitats with livestock and wild animals. We decided to investigate the role of dogs in the maintenance of Brucella spp. in the Pantanal wetland. Serum and whole blood samples were collected from 167 dogs. To detect antibodies against B. abortus and B. canis, buffered acidified plate antigen (BAPA) and agar gel immunodiffusion (AGID) tests were performed. To detect Brucella spp., B. abortus and B. canis DNA, PCR was performed using the bcsp31, BruAb2_0168, and BR00953 genes, respectively. To confirm the PCR results, three bcsp31 PCR products were sequenced and compared with sequences deposited in GenBank. The seropositivity rates of 7.8% and 9% were observed for the AGID and BAPA tests, respectively. Positivity rates of 45.5% and 10.8% were observed when testing bcsp31 and BruAb2_0168, respectively, while there was no positivity for BR00953. The sequenced products had 110 base pairs that aligned with 100% identity to B. abortus, B. canis, and B. suis. Considering our results, dogs may be acting as maintenance hosts of Brucella spp. in the Pantanal region.
Assuntos
Brucella abortus/isolamento & purificação , Brucella canis/isolamento & purificação , Brucelose/veterinária , Doenças do Cão/microbiologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Brucella abortus/genética , Brucella abortus/metabolismo , Brucella canis/genética , Brucella canis/metabolismo , Brucelose/microbiologia , Cães , Feminino , Masculino , Áreas AlagadasRESUMO
Brucella canis, un patógeno intracelular facultativo, es responsable de la brucelosis canina, una enfermedad zoonótica que afecta a los caninos y al hombre. En los primeros causa abortos y fallas reproductivas; en el ser humano genera síntomas inespecíficos. En el año 2005 se demostró la presencia de B. canis en Antioquia (Colombia). Las cepas halladas se identificaron como tipo 2. La secuenciación del genoma completo de una cepa de campo denominada Brucella canis str. Oliveri mostró indels específicos de especie; a partir de estos se buscó conocer características genómicas de las cepas de B. canis aisladas y establecer relaciones filogenéticas, así como el tiempo de divergencia de la cepa Oliveri. Se realizó PCR convencional y secuenciación de 30 cepas de campo, se identificaron 5 indels reconocidos en B. canis str. Oliveri, se empleó ADN de Brucella suis, Brucella melitensis y cepas vacunales de Brucella abortus como controles. Se determinó que las cepas de campo estudiadas comparten 4 de los 5 indels de la cepa Oliveri, lo que indica la presencia de más de una cepa de B. canis circulando en la región. El análisis filogenético se realizó con 24 cepas de Brucella mediante secuencias concatenadas de genes marcadores de especie. Se probó la hipótesis del reloj molecular y adicionalmente se realizó test de tasas relativas de Tajima. De esta manera se demostró que la cepa Oliveri, al igual que las otras cepas de B. canis analizadas, divergen de B. suis. Se rechazó la hipótesis del reloj molecular entre las especies de Brucella y se demostró una tasa de evolución y una distancia genética similar entre las cepas de B. canis.
Brucella canis is a facultative intracellular pathogen responsible for canine brucellosis, a zoonotic disease that affects canines, causing abortions and reproductive failure; and the production of non-specific symptoms in humans. In 2005 the presence of B. canis in Antioquia was demonstrated and the strains were identified as type 2. The sequencing of the genome of a field strain denoted Brucella canis str. Oliveri, showed species-specific indel events, which led us to investigate the genomic characteristics of the B. canis strain isolated and to establish the phylogenetic relationships and the divergence time of B. canis str. Oliveri. Conventional PCR sequencing was performed in 30 field strains identifying 5 indel events recognized in B. canis str. Oliveri. ADN from Brucella suis, Brucella melitensis and vaccine strains from Brucella abortus were used as control, and it was determined that all of the studied field strains shared 4 out of the 5 indels of the sequenced Oliveri strain, indicating the presence of more than one strain circulating in the region. Phylogenetic analysis was performed with 24 strains of Brucella using concatenated sequences of genetic markers for species differentiation. The molecular clock hypothesis and Tajima's relative rate test were tested, showing that the Oliveri strain, similarly to other canis species, diverged from B. suis. The molecular clock hypothesis between Brucella species was rejected and an evolution rate and a similar genetic distance between the B. canis were demonstrated.