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Master's Dissertation
DOI
https://doi.org/10.11606/D.10.2020.tde-18052020-102343
Document
Author
Full name
Henrique Tobaro Macedo
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Pirassununga, 2020
Supervisor
Committee
Brunetto, Marcio Antonio (President)
Jeremias, Juliana Toloi
Souza, Luciano Melo de
Title in Portuguese
Caracterização da microbiota fecal de cães obesos e após emagrecimento
Keywords in Portuguese
Canino
Disbiose
Microbioma
Obesidade
Trato gastrointestinal
Abstract in Portuguese
A obesidade é considerada na medicina veterinária a afecção nutricional e metabólica mais comum da atualidade. Esta ocorre devido ao acúmulo excessivo de tecido adiposo no organismo, resultado do aumento na ingestão de energia associado a falta de atividade física. A obesidade vem sendo correlacionada com disbiose da microbiota intestinal, bem como em desordens imunológicas, alterações ortopédicas, respiratórias e metabólicas, como resistência insulínica e hiperlipidemia. Estudos demonstraram em ratos geneticamente obesos e induzidos pela dieta, que a microbiota cecal de animais acima do peso apresentam tags genéticos ambientais que codificam enzimas envolvidas na degradação de polissacarídeos e no metabolismo de carboidratos simples. Assim, a microbiota de indivíduos obesos pode ser mais eficiente em digerir e metabolizar a energia da dieta que indivíduos magros. Dessa forma, o objetivo deste trabalho foi caracterizar a microbiota fecal de cães obesos e em escore corporal ideal e avaliar os efeitos do programa de perda de peso sobre os filos e gêneros bacterianos. Foram incluídos vinte cães, fêmeas, castradas, de diferentes raças, com idade entre 1 a 9 anos. Os animais foram divididos em dois grupos: grupo experimental de animais obesos (GO), com escore de condição corporal (ECC) 9 e com porcentagem de gordura corporal superior a 30%, determinada pelo método de diluição de isótopos de deutério e um grupo controle (GC), com ECC 4 ou 5 e gordura corporal máxima de 15%. Os animais obesos foram incluídos em um programa de perda de peso e, quando perderam 20% do peso inicial, passaram a compor o grupo de animais emagrecidos (GE). Foram coletadas amostras de fezes dos três grupos experimentais. Nos grupos controle e obesos, a coleta foi realizada após um período de adaptação com dieta de manutenção e, no grupo emagrecido, a coleta de fezes foi realizada no final do programa de perda de peso. Após cada coleta, as amostras foram armazenadas em freezer -80°C. Na sequência, foi extraído o DNA total das fezes e estas foram sequenciadas pela metodologia Illumina. As abundâncias observadas para cada filo e gênero foram avaliadas por modelo linear generalizado, considerando distribuição binominal e pelo emprego da função de ligação logit. O modelo incluiu efeitos fixos de grupos (Controle, Obesos e Emagrecidos), além dos efeitos aleatórios de animal e resíduo. Todas as análises foram realizadas pelo procedimento PROC GLIMMIX do programa Statistical Analysis System. Valores de P<0,05 foram considerados significativos. Foram identificados 7 filos e 51 gêneros diferentes entre os 3 grupos experimentais. O filo e gênero predominante entre os grupos foi respectivamente o Firmicutes (53,01% - 71,98%) e o Clostridium (42,43 25,51%). Foi observada diferença entre animais obesos e magros para o filo Firmicutes (P=0,0189) e gênero Sutturella (P=0,0095). O programa de perda de peso modulou de forma positiva os filos Actinobacteria (P=0,0128), Firmicutes (P=0,0189) e o gênero Dorea (P<,0001) e, negativamente o Faecalibacterium (P=0,0114). O presente trabalho verificou que o programa de perda de peso foi capaz de modular os microrganismos do trato gastrointestinal, de forma que, os cães emagrecidos apresentaram composição microbiana com maior biodiversidade, aumento de Actinobacteria, Firmicutes e Dorea e menor abundancia de Faecalibacterium.
Title in English
Characterization of fecal microbiota of obese dogs and after weight loss
Keywords in English
Canine
Dysbiosis
Gastrointestinal tract
Microbiome
Obesity
Abstract in English
Obesity is considered the most common nutritional and metabolic disorder in veterinary medicine today. This occurred to the excessive accumulation of adipose tissue in the organism, due to increase in energy intake with low physical activity. Obesity has been associated with an intestinal microbiota dysbiosis, as well as in immune disorders, orthopedic, respiratory and metabolic changes, such as insulin resistance and hyperlipidemia. Studies have shown in both genetically obese and diet-induced mice that the cecal microbiota from obese animals is enriched with environmental genetic tags that encode enzymes involved in the degradation of polysaccharides and the metabolism of simple carbohydrates. Thus, the microbiota of obese individuals may be more efficient in digesting and metabolizing dietary energy than lean individuals. So, the present study aims to characterize the microbioma profile of obese dogs and to evaluate the effects of weight loss on the population of microorganisms of dogs included in a weight loss program. Twenty dogs, female, neutered, different breeds and 1 to 9 years old were included. The animals were divided into two groups: experimental group of obese animals (GO) with a body condition score (BCS) 9 and body fat percentage greater than 30% determined by the deuterium isotope dilution method and a control group (GC) with BCS 4 or 5 and maximum body fat of 15%. The obese animals were included in a weight loss program and when they lost 20% of their initial weight, they became part of the group of emaciated animals (GE). Stool samples were collected from the three experimental groups. In the control and obese groups, the collection was performed after an adaptation period with a maintenance diet and in the emaciated group, the collection of feces was done at the end of the weight loss program. After each collection, the samples were stored at -80°C. Total DNA was extracted from the feces and sequenced using the Illumina methodology. The abundances observed for each phylum and gender were evaluated using a generalized linear model, considering binomial distribution and using the logit link function. The model includes fixed effects of Groups (Control, Obese and Weight Loss), in addition to the random effects of animals and residue. All as analyzes were performed using PROC GLIMMIX procedure from the Statistical Analysis System Program. Values of P<0.05 were considered significant. Seven different phyla and 51 different males were identified among the three experimental groups. The most prevalent phylum and genus among the groups was Firmicutes (53.01% - 71.98%) and Clostridium (42.43 25.51%) respectively. It was observed difference between obese and lean animals for phyla Firmicutes (P = 0.0189) and genus Sutturella (P = 0.0095). The weight loss program regulated positively the phyla Actinobacteria (P = 0.0128), Firmicutes (P = 0.0189) and genus Dorea (P P<0001), and negatively genus Faecalibacterium (P = 0.0114). The present study found that the weight loss program is capable of modulating the microorganisms of the gastrointestinal tract, so that the emaciated dogs presented microbial composition with greater biodiversity, greater presence of Actinobacteria, Firmicutes and Dorea and less abundance of Faecalibacterium.
 
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Publishing Date
2020-08-14
 
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