RESUMO
ABSTRACT: Clostridium chauvoei toxin A (CctA), neuraminidase (NanA), and flagellin (FliC) proteins contribute to the pathogenicity of Clostridium chauvoei, the causative agent of blackleg in cattle. The aim of this study was to analyze the genetic variability of cctA, nanA, and fliC genes in C. chauvoei isolates from the Rio Grande do Sul and São Paulo state- Brazil, during different sampling periods. The presence of these genes was verified through PCR amplification and partial gene sequencing of 17 strains. Alignment of PCR amplicons combined with bioinformatics analysis was used in an attempt to study the variability across C. chauvoei solates. The similarity among the partial sequences of cctA and nanA genes was 100%. The sequencing of fliC revealed three different paralog alleles of flagellin, and two strains were seen to be polymorphic, with amino acid alterations in the predicted protein. Overall, this study indicates that strains of C. chauvoei isolated in Brazil are highly conserved with respect to the virulence factors evaluated.
RESUMO: Toxina A de Clostridium chauvoei (CctA), neuraminidase (NanA) e flagelina (FliC) são proteínas que contribuem para a patogenicidade de Clostridium chauvoei, o agente causador do carbúnculo sintomático em bovinos. O objetivo deste estudo foi analisar a variabilidade genética dos genes cctA, nanA, e fliC em C. chauvoei isolados em diferentes períodos no Rio Grande do Sul e São Paulo. A presença destes genes foi verificada pela amplificação dos produtos da PCR e sequenciamento parcial dos genes de 17 cepas. Os alinhamentos da amplificação dos produtos da PCR combinados com a análise de bioinformática foram utilizados na tentativa de avaliar a variabilidade dos genes entre os isolados de C. chauvoei. A similaridade do sequenciamento parcial dos genes cctA e nanA foi 100%. O sequenciamento do fliC revelou três alelos paralogos diferentes de flagelina e duas cepas mostraram polimorfismos, causando alterações na sequência de aminoácidos. As cepas de C. chauvoei isoladas no Brasil mostraram-se altamente conservadas em relação aos fatores de virulência avaliados neste estudo.
RESUMO
The cytotoxin A (MbxA) is one of the main virulence factors of Moraxella bovis involved in the pathogenesis of infectious bovine keratoconjunctivitis (IBK). Moraxella ovis and Moraxella bovoculi, suspected to be associated with infectious keratitis in sheep and cattle respectively, also have a gene that encodes the cytotoxin A (movA and mbvA, respectively). The aim of this study was to determine the molecular sequence of the 3' region of the cytotoxin gene of Moraxella spp. strains isolated from clinical cases to establish phylogenetic and evolutionary comparisons. PCR amplification, nucleotide sequencing (nt) and amino acid (aa) sequence prediction were performed, followed by the sequences comparison, identity level calculation and selective pressure analysis. The phylogenetic reconstruction based on nt and aa sequences clearly differentiate M. bovis (n=15), M. bovoculi (n=11) and M. ovis (n=7) and their respective reference strains. An alignment of 843nt revealed high similarity within bacterial species (MbxA=99.9% nt and aa; MbvA=99.3% nt and 98.8% aa; MovA=99.5% nt and 99.3% aa). The similarity of partial sequences (nt 1807-2649) of MbxA in relation to MbvA and MovA ranged from 76.3 to 78.5%; similarity between MbvA and MovA ranged from 95.7 to 97.5%. A negative selection on mbvA and movA sequences was revealed by the molecular evolution analysis. The phylogenetic analysis of movA and mbvA allowed different strains of Moraxella spp. to be grouped according to the period of isolation. Sequence analysis of cytotoxin may provide insights into genetic and evolutionary relationships and into the genetic/molecular basis of Moraxella spp.
Assuntos
Doenças dos Bovinos/microbiologia , Variação Genética , Ceratoconjuntivite Infecciosa/microbiologia , Moraxella/genética , Infecções por Moraxellaceae/veterinária , Doenças dos Ovinos/microbiologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Citotoxinas/genética , Geografia , Dados de Sequência Molecular , Moraxella/isolamento & purificação , Moraxella bovis/genética , Moraxella bovis/isolamento & purificação , Infecções por Moraxellaceae/microbiologia , Filogenia , Análise de Sequência de DNA , OvinosRESUMO
The objective of this study was to describe the first report involving a case of equine acute myonecrosis caused by C. novyi type A with an emphasis on clinical signs, the pathological and bacteriological analysis, and molecular identification of the microorganisms as the key of the definitive diagnosis.
Assuntos
Infecções por Clostridium/veterinária , Clostridium/classificação , Clostridium/isolamento & purificação , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/patologia , Miosite/veterinária , Necrose/veterinária , Animais , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/microbiologia , Infecções por Clostridium/patologia , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Histocitoquímica , Doenças dos Cavalos/microbiologia , Cavalos , Miosite/diagnóstico , Miosite/microbiologia , Miosite/patologia , Necrose/diagnóstico , Necrose/microbiologia , Necrose/patologia , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
The objective of this study was to describe the first report involving a case of equine acute myonecrosis caused by C. novyi type A with an emphasis on clinical signs, the pathological and bacteriological analysis, and molecular identification of the microorganisms as the key of the definitive diagnosis.
Assuntos
Animais , Infecções por Clostridium/veterinária , Clostridium/classificação , Clostridium/isolamento & purificação , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/patologia , Miosite/veterinária , Necrose/veterinária , Análise por Conglomerados , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/microbiologia , Infecções por Clostridium/patologia , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Histocitoquímica , Cavalos , Doenças dos Cavalos/microbiologia , Miosite/diagnóstico , Miosite/microbiologia , Miosite/patologia , Necrose/diagnóstico , Necrose/microbiologia , Necrose/patologia , Filogenia , /genética , Análise de Sequência de DNARESUMO
This study aimed to evaluate and select different dermatological bases incorporated with propolis for veterinary use as well as to analyze the chemical compounds of the propolis hydroalcoholic extract by LC-MS/MS. Thus, formulations were submitted to accelerated stability tests under different temperatures and to mechanical stress, and evaluated for the appearance, color, odor, pH, viscosity, spreadability, and the mean size of the dispersed globules from the internal phase during a period of three months. The creamy gel formulation showed satisfactory results for all the evaluated items with an excellent capability to incorporate the hydroalcoholic extract of propolis associated to the maintenance of its physicochemical properties. The propolis used in this study had been shown to possess antibacterial and antifungal in vitro activity against the main microorganisms responsible for such diseases. Therefore, the propolis creamy gel described here could be a promising formulation for use in the veterinary medicine.
