Genotipagem de bactérias anaeróbias associadas às lesões da periodontite bovina
Data
2017-02-07
Autores
Borsanelli, Ana Carolina [UNESP]
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
A periodontite bovina é um processo infeccioso purulento e progressivo associado com a presença de biofilme subgengival anaeróbio. De caráter sazonal e associada ao manejo do solo e à dieta, a doença tem variações na sua apresentação clínica, que inclui desde uma forma agressiva até manifestações crônicas. Os prejuízos econômicos são significativos e decorrem das dificuldades na preensão, mastigação e ruminação. O presente estudo teve como objetivo geral caracterizar a periodontite bovina e identificar por métodos moleculares os micro-organismos associados às lesões periodontais. Na avaliação da microbiota da bolsa periodontal (n=26) e do sulco subgengival (n=25) de bovinos, pela reação em cadeia da polimerase (PCR) e com o emprego de 35 iniciadores de espécies de patógenos potenciais, pôde-se associar a ocorrência de Actinobacillus naeslundii, Enterococcus faecium, Porphyromonas asaccharolytica, Porphyromonas endodontalis, Prevotella buccae, Prevotella intermedia, Prevotella melaninogenica, Prevotella nigrescens, Prevotella oralis, Treponema denticola e Treponema pectinovorum com a periodontite bovina. Em estudo complementar realizado na Escócia para verificar a ocorrência de lesões periodontais em animais abatidos, foram examinadas 200 arcadas dentárias, das quais 24 (12%) apresentaram lesões periodontais nos dentes incisivos ou mastigatórios. Este estudo inédito revela que a periodontite não é incomum em bovinos abatidos no oeste da Escócia e é claramente um problema sanitário negligenciado na produção e bem-estar animal. Na oportunidade, os fatores de risco associados à doença foram avaliados em um universo de 250 animais abatidos, dos quais 35 apresentavam lesões periodontais e 40 eram periodontalmente sadios. Pela análise de regressão logística foi avaliada a associação entre as variáveis independentes, sexo, idade e raça com periodontite. A idade dos animais foi significativamente associada à presença de lesões periodontais. Para cada ano de idade, um bovino tem 1,53 vezes chances de desenvolver periodontite (p<0,001). A variável sexo não se mostrou significativamente associada à periodontite (p=0,887), enquanto os animais de corte têm 0,36 a chance de desenvolver a doença quando comparados com os de aptidão leiteira. Na mesma ocasião, amostras de biofilme subgengival de 40 bovinos com periodontite e de 38 periodontalmente sadios foram coletadas e realizou-se o sequenciamento do gene 16S rRNA. No microbioma de animais sadios os gêneros mais prevalentes foram Gastranaerophilus, Planifilus, Burkholderia e Arcobacter. Já nos animais com periodontite, os filos mais prevalentes foram Elusimicrobia, Synergista e Fusobacteria e os gêneros Propionivibrio, Wolinella, Porphyromonas, Candidatus, Prevotella, Firmicutes (não cultivável), Bacteroides e Treponema. Em conclusão, os dois grupos de bovinos avaliados abrigaram perfis microbianos distintos, sendo que as amostras de bovinos com periodontite foram mais diversas em micro-organismos do que as de bovinos sadios. Nesse contexto inédito na microbiologia oral de bovinos, pode-se constatar os componentes principais na homeostase bacteriana do biofilme de sítios sadios e a disbiose nas lesões periodontais, fornecendo indicadores para e evolução do conhecimento sobre a periodontite bovina.
Bovine periodontitis is a progressive purulent infectious process associated with the presence of strict anaerobic subgingival biofilm. Seasonal and associated to soil and dietary management, the disease has variations in its clinical presentation, which includes since an aggressive form until chronic manifestations. The economic losses are significant and result from difficulties in gripping, chewing and rumination. The present study aimed to identify and characterize bovine periodontitis and identify the microorganisms associated with periodontal lesions by molecular methods. In the evaluation of the microbiota of the periodontal pocket (n=26) and gingival sulcus (n=25) of cattle, by polymerase chain reaction (PCR) and with the use of 35 primers of species of potential pathogens, it can be associate the occurrence of Actinobacillus naeslundii, Enterococcus faecium, Porphyromonas asaccharolytica, Porphyromonas endodontalis, Prevotella buccae, Prevotella intermedia, Prevotella melaninogenica, Prevotella nigrescens, Prevotella oralis, Treponema denticola and Treponema pectinovorum with bovine periodontitis. In a study carried out in Scotland to verify the occurrence of periodontal lesions in slaughtered animals, 200 dental arches were examined, of which 24 (12%) presented periodontal lesions in the incisors or masticatory teeth. This unpublished study reveals that periodontitis is not uncommon in cattle slaughtered in West of Scotland and is clearly a neglected health problem in animal production and welfare. At the opportunity, the risk factors associated with the disease were evaluated in a universe of 250 slaughtered animals, of which 35 had periodontal lesions and 40 were periodontally healthy. By the logistic regression analysis was evaluated the association between the independent variables, sex, age and race with periodontitis. The age of the animals was significantly associated with the presence of periodontal lesions. For each extra year in age, a cow is 1.53 times more likely to develop periodontitis (p<0.001). Gender was not significantly associated with periodontitis (p=0.887). Regarding the variable breed type, beef cattle were 0.36 times as likely to have periodontitis compared to dairy cattle. At the same occasion, samples of subgengival biofilm of 40 bovines with periodontitis and 38 periodontally healthy were submitted to high- throughput sequencing. In the bovine microbiome the most discriminative taxa in the samples of healthy animals were Gastranaerophilus, Planifilus, Burkholderia and Arcobacter. In animals with periodontitis, the most prevalent microorganisms were Elusimicrobia, Synergista, Propionivibrio, Fusobacteria, Wolinella, Porphyromonas, Candidatus, Prevotella, Firmicutes (uncultivable), Bacteroides and Treponema. In conclusion, the two groups of bovines evaluated harboured distinct microbial profiles, and the samples of bovines with periodontitis were more diverse in microorganisms than those of healthy cattle. In this unprecedented context, in the oral microbiology of bovines we can verify the main components in the bacterial homeostasis of the biofilm of healthy sites and the dysbiosis in the periodontal lesions, providing indicators for and evolution of the knowledge on bovine periodontitis.
Bovine periodontitis is a progressive purulent infectious process associated with the presence of strict anaerobic subgingival biofilm. Seasonal and associated to soil and dietary management, the disease has variations in its clinical presentation, which includes since an aggressive form until chronic manifestations. The economic losses are significant and result from difficulties in gripping, chewing and rumination. The present study aimed to identify and characterize bovine periodontitis and identify the microorganisms associated with periodontal lesions by molecular methods. In the evaluation of the microbiota of the periodontal pocket (n=26) and gingival sulcus (n=25) of cattle, by polymerase chain reaction (PCR) and with the use of 35 primers of species of potential pathogens, it can be associate the occurrence of Actinobacillus naeslundii, Enterococcus faecium, Porphyromonas asaccharolytica, Porphyromonas endodontalis, Prevotella buccae, Prevotella intermedia, Prevotella melaninogenica, Prevotella nigrescens, Prevotella oralis, Treponema denticola and Treponema pectinovorum with bovine periodontitis. In a study carried out in Scotland to verify the occurrence of periodontal lesions in slaughtered animals, 200 dental arches were examined, of which 24 (12%) presented periodontal lesions in the incisors or masticatory teeth. This unpublished study reveals that periodontitis is not uncommon in cattle slaughtered in West of Scotland and is clearly a neglected health problem in animal production and welfare. At the opportunity, the risk factors associated with the disease were evaluated in a universe of 250 slaughtered animals, of which 35 had periodontal lesions and 40 were periodontally healthy. By the logistic regression analysis was evaluated the association between the independent variables, sex, age and race with periodontitis. The age of the animals was significantly associated with the presence of periodontal lesions. For each extra year in age, a cow is 1.53 times more likely to develop periodontitis (p<0.001). Gender was not significantly associated with periodontitis (p=0.887). Regarding the variable breed type, beef cattle were 0.36 times as likely to have periodontitis compared to dairy cattle. At the same occasion, samples of subgengival biofilm of 40 bovines with periodontitis and 38 periodontally healthy were submitted to high- throughput sequencing. In the bovine microbiome the most discriminative taxa in the samples of healthy animals were Gastranaerophilus, Planifilus, Burkholderia and Arcobacter. In animals with periodontitis, the most prevalent microorganisms were Elusimicrobia, Synergista, Propionivibrio, Fusobacteria, Wolinella, Porphyromonas, Candidatus, Prevotella, Firmicutes (uncultivable), Bacteroides and Treponema. In conclusion, the two groups of bovines evaluated harboured distinct microbial profiles, and the samples of bovines with periodontitis were more diverse in microorganisms than those of healthy cattle. In this unprecedented context, in the oral microbiology of bovines we can verify the main components in the bacterial homeostasis of the biofilm of healthy sites and the dysbiosis in the periodontal lesions, providing indicators for and evolution of the knowledge on bovine periodontitis.
Descrição
Palavras-chave
Periodontite, Bovinos, Anaeróbios, PCR, Sequenciamento de alto rendimento, Fatores de risco, Periodontitis, Bovine, Anaerobics, High-throughput sequencing, Risk factors