A influência da 2’-fucosil-lactose, galacto-oligossacarídeo e fruto-oligossacarídeo na microbiota intestinal de lactentes saudáveis
Data
2024-03-01
Autores
Lazarini, Tamara 
Orientadores
Morais, Mauro Batista de
Tipo
Tese de doutorado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Objetivo: Os oligossacarídeos do leite humano (HMOs) são considerados o segundo carboidrato mais abundante do leite humano e com impacto positivo no estabelecimento de uma microbiota intestinal infantil saudável. Analisar a influência dos oligossacarídeos 2´-FL (2-Fucosil-Lactose), GOS (Galacto-oligossacarídeo) e FOS (Fruto-oligossacarídeo), na modulação da microbiota intestinal de lactentes saudáveis em aleitamento artificial foi o principal objetivo do estudo. Métodos: Foram examinadas fezes de 87 lactentes no 1º. e 4º. mês de vida: grupo LM (n=28) em aleitamento materno como referência, grupo HMO (n=29) alimentados com a fórmula experimental com 2´-FL+GOS+FOS e o grupo GOS/FOS (n=30) alimentados com a fórmula controle com GOS+FOS. As amostras fecais foram analisadas utilizando o kit DNA/RNA Shield R1101 e a extração de DNA foi realizada utilizando o kit ZymoBIOMICS DNA Miniprep. A região bacteriana V3 e V4 do gene 16S rRNA foi amplificada e sequenciada no MiSeq Illumina. As sequências geradas foram analisadas utilizando o software QIIME2. Os testes de variância ANOVA, Kruskal-Wallis, índices de riqueza Chao1 e Shannon, as distâncias UniFrac e o teste de Adonis foram utilizados para as diferentes análises estatísticas. Para as vertentes clínicas foram consideradas as variáveis antropométricas e dados dos questionários autopreenchidos. Resultados: O Actinobacteriota foi o filo mais prevalente nos grupos HMO (60,4%) e LM (46,6%), e o Firmicutes apresentou maior abundância relativa para o grupo GOS/FOS (42,4%). A Bifidobacterium e a Escherichia-Shigella foram os dois gêneros bacterianos mais abundantes ao final do estudo nos três grupos. Os lactentes alimentados com fórmulas apresentou maior abundância relativa dos gêneros [Ruminococcus]_gnavus_group. O grupo LM apresentou maior abundância relativa do gênero Lacticaseibacillus em relação ao grupo GOS/FOS. Os grupos que receberam aleitamento artificial apresentaram maior alfa-diversidade da microbiota intestinal quando comparado aos lactentes amamentados (p>0,05). A análise de beta-diversidade não mostrou diferença segundo o tipo de parto ou sexo dos lactentes, no entanto, apresentou diferença estatisticamente significante segundo o tipo de alimentação por grupo, na matriz UNIFRAC ponderada (p< 0,004) e não-ponderada (p< 0,001). Todos os lactentes tiveram crescimento adequado para a idade, sem efeitos adversos. Conclusões: A adição de oligossacarídeos (2’-FL, GOS e FOS) nas fórmulas infantis não aproximou a alfa diversidade da microbiota intestinal dos lactentes em aleitamento artificial à do referencial do leite materno. A beta-diversidade dos três grupos (HMO, GOS+FOS e LM) foram diferentes entre si no final do estudo. A adição da molécula de HMO (2´-FL) em uma fórmula infantil com a combinação de prebióticos GOS+FOS, proporcionou maior abundância relativa do filo Actinobacteriota e do gênero Bifidobacterium ao final da intervenção, que foi similar ao grupo do leite materno (p>0,05). Do ponto de vista clínico, se associou com menor tempo de irritação e mais horas de sono por noite em relação aos demais grupos.
Objective: Human milk oligosaccharides (HMOs) are considered the second most abundant carbohydrate in human milk and have a positive impact on the establishment of a healthy infant gut microbiota. Analyzing the influence of the oligosaccharides 2'-FL (2-Fucosyl-Lactose), GOS (Galacto-oligosaccharide) and FOS (Fructo-oligosaccharide), in modulating the gut microbiota of healthy infants on artificial feeding was the main objective of the study. Methods: Feces from 87 infants were examined in the 1st. and 4th. month of life: LM group (n=28) breastfed infants as reference, HMO group (n=29) fed with the experimental formula with 2´-FL+GOS+FOS and the GOS/FOS group (n=30) fed with the control formula with GOS+FOS. Fecal samples were analyzed using the DNA/RNA Shield R1101 kit and DNA extraction was performed using the ZymoBIOMICS DNA Miniprep kit. The bacterial V3 and V4 region of the 16S rRNA gene was amplified and sequenced on MiSeq Illumina. The generated sequences were analyzed using QIIME2 software. The ANOVA, Kruskal-Wallis, Chao1 & Shannon richness indices, UniFrac distances and the Adonis test were used for the different statistical analyses. For the clinical aspects, anthropometric variables and data from self-completed questionnaires were considered. Results: Actinobacteriota was the most prevalent phylum in the HMO (60.4%) and LM (46.6%) groups, and Firmicutes presented the highest relative abundance for the GOS/FOS group (42.4%). Bifidobacterium and Escherichia-Shigella were the two most abundant bacterial genera at the end of the study in the three groups. Formula-fed infants had a higher relative abundance of the [Ruminococcus]_gnavus_group genera. The LM group showed a higher relative abundance of the genus Lacticaseibacillus compared to the GOS/FOS group. The groups that received formula feeding demonstrated higher alpha-diversity of the intestinal microbiota when compared to breastfed infants (p>0.05). The beta-diversity analysis showed no difference according to the type of delivery or sex of the infants; however, it showed a statistically significant difference according to the type of feeding per group, considering the weighted (p< 0.004) and unweighted (p< 0.001) UNIFRAC matrix. All infants had adequate growth for age, without adverse symptoms. Conclusions: The addition of oligosaccharides (2'-FL, GOS and FOS) to infant formulas did not resemble the gut microbiota alpha-diversity of formula fed infants closer to that the breastfeeding infants. The beta-diversity of the three groups (HMO, GOS+FOS and LM) were different from each other at the end of the study. The addition of 2'-FL (HMO molecule) to an infant formula with the combination of GOS+FOS prebiotics, provided a greater relative abundance of the phylum Actinobacteriota and the genus Bifidobacterium at the end of the intervention, which was similar to the breastfeeding infants group (p>0.05). From a clinical point of view, it was associated with less irritation time and more hours of sleep per night compared to the other groups.
Objective: Human milk oligosaccharides (HMOs) are considered the second most abundant carbohydrate in human milk and have a positive impact on the establishment of a healthy infant gut microbiota. Analyzing the influence of the oligosaccharides 2'-FL (2-Fucosyl-Lactose), GOS (Galacto-oligosaccharide) and FOS (Fructo-oligosaccharide), in modulating the gut microbiota of healthy infants on artificial feeding was the main objective of the study. Methods: Feces from 87 infants were examined in the 1st. and 4th. month of life: LM group (n=28) breastfed infants as reference, HMO group (n=29) fed with the experimental formula with 2´-FL+GOS+FOS and the GOS/FOS group (n=30) fed with the control formula with GOS+FOS. Fecal samples were analyzed using the DNA/RNA Shield R1101 kit and DNA extraction was performed using the ZymoBIOMICS DNA Miniprep kit. The bacterial V3 and V4 region of the 16S rRNA gene was amplified and sequenced on MiSeq Illumina. The generated sequences were analyzed using QIIME2 software. The ANOVA, Kruskal-Wallis, Chao1 & Shannon richness indices, UniFrac distances and the Adonis test were used for the different statistical analyses. For the clinical aspects, anthropometric variables and data from self-completed questionnaires were considered. Results: Actinobacteriota was the most prevalent phylum in the HMO (60.4%) and LM (46.6%) groups, and Firmicutes presented the highest relative abundance for the GOS/FOS group (42.4%). Bifidobacterium and Escherichia-Shigella were the two most abundant bacterial genera at the end of the study in the three groups. Formula-fed infants had a higher relative abundance of the [Ruminococcus]_gnavus_group genera. The LM group showed a higher relative abundance of the genus Lacticaseibacillus compared to the GOS/FOS group. The groups that received formula feeding demonstrated higher alpha-diversity of the intestinal microbiota when compared to breastfed infants (p>0.05). The beta-diversity analysis showed no difference according to the type of delivery or sex of the infants; however, it showed a statistically significant difference according to the type of feeding per group, considering the weighted (p< 0.004) and unweighted (p< 0.001) UNIFRAC matrix. All infants had adequate growth for age, without adverse symptoms. Conclusions: The addition of oligosaccharides (2'-FL, GOS and FOS) to infant formulas did not resemble the gut microbiota alpha-diversity of formula fed infants closer to that the breastfeeding infants. The beta-diversity of the three groups (HMO, GOS+FOS and LM) were different from each other at the end of the study. The addition of 2'-FL (HMO molecule) to an infant formula with the combination of GOS+FOS prebiotics, provided a greater relative abundance of the phylum Actinobacteriota and the genus Bifidobacterium at the end of the intervention, which was similar to the breastfeeding infants group (p>0.05). From a clinical point of view, it was associated with less irritation time and more hours of sleep per night compared to the other groups.
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Citação
LAZARINI, Tamara. A influência da 2’-fucosil-lactose, galacto-oligossacarídeo e fruto-oligossacarídeo na microbiota intestinal de lactentes saudáveis. 2024. 170 f. Tese (Doutorado) - Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, 2024.