Proteoma salivar e microbiomas dental e ruminal de bovinos mantidos em dieta com suplemento proteico
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Data
2022-12-06
Autores
Saraiva, Júlia Rebecca [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A boca é um ambiente ecologicamente dinâmico composto por elementos estruturais que lhe conferem uma identidade fisiológica específica, como a deposição de biofilmes dentais e a expressão de proteínas salivares associadas a processos locais e sistêmicos. As doenças periodontais afetam bovinos e outros ruminantes causando impactos na produção e bem-estar animal, e acredita-se que sua etiologia esteja associada ao manejo alimentar dos animais. O objetivo do presente estudo foi avaliar o efeito de dieta suplementar proteica e feno de Panicum cultivado em áreas recém-reformadas no proteoma salivar e integridade periodontal, e microbioma dental e ruminal de bezerros em fase de dentição decídua. Foram utilizados 12 bovinos divididos aleatoriamente em dois grupos: dieta base feno (F) e dieta suplementar proteica (SP) durante 60 dias. A saliva foi coletada aos 20 e 60 dias para análise proteômica por espectrometria de massas nanoAcquity UPLC-Xevo QTof MS, software ProteinLynx Global Server (PLGS) e o número de acesso as proteínas foi fornecido pelo banco de dados UniProt para espécie Bos taurus. O exame clínico bucal foi realizado semanalmente pela inspeção da cavidade bucal e sondagem periodontal dos incisivos. Já para o sequenciamento, coletou-se biofilme subgengival e 15mL de conteúdo ruminal no início e término do experimento (0 e 60 dias) para análise de sequenciamento de última geração do gene 16S rRNA. A análise de interação entre tempo e dieta demonstrou significância na incidência de episódios de gengivite entre os dois grupos aos 20 dias e aos 60 e 20 dias da dieta exclusiva de feno. Aos 60 dias da experimentação, mais processos biológicos (9) e imunológicos (11) estiveram alterados na saliva dos bovinos. Proteínas com múltiplas funções foram detectadas na saliva dos animais, principalmente de função antimicrobiana e anti-inflamatória. No microbioma dental, Filos já reconhecidos em bovinos foram identificados, como Proteobacteria, Bacteroidetes, Fusobateria. Os gêneros Moraxella, Neisseria e Lautropia destacaram-se em ambos os momentos. No rúmen, Filos considerados centrais dentro desse ambiente como Bacteroidetes e Firmicutes foram os mais representativos. Não foram observadas diferenças significativas de riqueza e diversidade do microbioma bucal e ruminal dos animais, entretando houve dissimilaridade na composição do microbioma bucal dos bovinos que receberam dieta base feno entre período inicial e final (p= 0.007), e no microbioma ruminal no grupo dieta base feno (p= 0.004) e nos bovinos do grupo dieta suplementar proteica (comparações entre os períodos inicial e final, p= 0.004) bem como entre as duas dietas no momento final (p= 0.005). Aparentemente, dietas com suplemento proteico estimulam a expressão de proteínas salivares com papel protetivo em bovinos, e interferem na modulação dos microbiomas (que ocorre inicialmente no rúmen) e provavelmente refletirá na boca dos animais.
The mouth is an ecologically dynamic environment composed of structural elements that confer a specific physiological identity to this location, such as the deposition of dental biofilms and the expression of salivary proteins associated with local and systemic processes. Periodontal diseases affect cattle and other ruminants causing impacts on animal production and welfare, and its etiology is believed to be associated with the feeding management of animals. The aim of the present study was to evaluate the effect of supplemental protein diet and Panicum hay grown in newly reformed areas on the salivary proteome and periodontal integrity, and dental and ruminal microbiome of calves in the deciduous phase. Twelve cattle were randomLy divided into two groups: hay-based diet (F) and protein supplemental diet (SP) for 60 days. Saliva was collected at 20 and 60 days for proteomic analysis by nanoAcquity UPLC-Xevo QTof MS mass spectrometry, ProteinLynx Global Server (PLGS) software and the protein accession number was provided by the UniProt database for Bos taurus species. The oral clinical examination was performed weekly by inspection of the oral cavity and periodontal probing of the incisors. For sequencing, subgingival biofilm and 15mL of rumen content were collected at the beginning and end of the experiment for analysis of next-generation sequencing of the 16S rRNA gene. The analysis of interaction between time and diet showed a significance in the incidence of gingivitis episodes between the two groups at 20 days and at 60 and 20 days of the hay-only diet. At 60 days of experimentation, more biological (9) and immunological (11) processes were altered in bovine saliva. Proteins with multiple functions were detected in the animals' saliva, mainly with antimicrobial and anti-inflammatory functions. In the dental microbiome, phyla already recognized in cattle were identified, such as Proteobacteria, Bacteroidetes, Fusobateria, and at the genus level Moraxella, Neisseria and Lautropia stood out in both moments. In the rumen, phyla considered central within this environment such as Bacteroidetes and Firmicutes were the most representative. In the rumen, phyla considered central within this environment such as Bacteroidetes and Firmicutes were the most representative. There were no significant differences in the richness and diversity of the oral and ruminal microbiome of the animals, however there was dissimilarity in the composition of the oral microbiome of cattle that received a haybased diet between the initial and final period (p= 0.007), and in the ruminal microbiome in the diet group hay base (p= 0.004) and in the cattle of the supplementary protein diet group (comparisons between the initial and final periods, p= 0.004) as well as between the two diets at the final moment (p= 0.005). Apparently, protein-supplemented diets stimulate the expression of salivary proteins with a protective role in cattle, and interfere with the modulation of microbiomes (which initially occurs in the rumen) and will probably be reflected in the animals' mouths.
The mouth is an ecologically dynamic environment composed of structural elements that confer a specific physiological identity to this location, such as the deposition of dental biofilms and the expression of salivary proteins associated with local and systemic processes. Periodontal diseases affect cattle and other ruminants causing impacts on animal production and welfare, and its etiology is believed to be associated with the feeding management of animals. The aim of the present study was to evaluate the effect of supplemental protein diet and Panicum hay grown in newly reformed areas on the salivary proteome and periodontal integrity, and dental and ruminal microbiome of calves in the deciduous phase. Twelve cattle were randomLy divided into two groups: hay-based diet (F) and protein supplemental diet (SP) for 60 days. Saliva was collected at 20 and 60 days for proteomic analysis by nanoAcquity UPLC-Xevo QTof MS mass spectrometry, ProteinLynx Global Server (PLGS) software and the protein accession number was provided by the UniProt database for Bos taurus species. The oral clinical examination was performed weekly by inspection of the oral cavity and periodontal probing of the incisors. For sequencing, subgingival biofilm and 15mL of rumen content were collected at the beginning and end of the experiment for analysis of next-generation sequencing of the 16S rRNA gene. The analysis of interaction between time and diet showed a significance in the incidence of gingivitis episodes between the two groups at 20 days and at 60 and 20 days of the hay-only diet. At 60 days of experimentation, more biological (9) and immunological (11) processes were altered in bovine saliva. Proteins with multiple functions were detected in the animals' saliva, mainly with antimicrobial and anti-inflammatory functions. In the dental microbiome, phyla already recognized in cattle were identified, such as Proteobacteria, Bacteroidetes, Fusobateria, and at the genus level Moraxella, Neisseria and Lautropia stood out in both moments. In the rumen, phyla considered central within this environment such as Bacteroidetes and Firmicutes were the most representative. In the rumen, phyla considered central within this environment such as Bacteroidetes and Firmicutes were the most representative. There were no significant differences in the richness and diversity of the oral and ruminal microbiome of the animals, however there was dissimilarity in the composition of the oral microbiome of cattle that received a haybased diet between the initial and final period (p= 0.007), and in the ruminal microbiome in the diet group hay base (p= 0.004) and in the cattle of the supplementary protein diet group (comparisons between the initial and final periods, p= 0.004) as well as between the two diets at the final moment (p= 0.005). Apparently, protein-supplemented diets stimulate the expression of salivary proteins with a protective role in cattle, and interfere with the modulation of microbiomes (which initially occurs in the rumen) and will probably be reflected in the animals' mouths.
Descrição
Palavras-chave
Bovinos, Dieta, Periodontal diseases, Proteomics, High-Throughput Nucleotide Sequencing