Análise da microbiota do colostro de vacas pluríparas e primíparas e de fezes de vacas e bezerros leiteiros no pós parto
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2018-12-06
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Universidade Federal de Viçosa
Resumo
Neste trabalho, o objetivo foi avaliar a composição da microbiota do colostro e fezes de vacas pluríparas e primíparas, assim como das fezes dos bezerros no pós- parto. As análises da microbiota não cultivada do colostro (1, 2 e 3 dias após o parto) e das fezes das vacas e dos bezerros foram realizadas um dia após o parto utilizando sequenciamento de nova geração do rRNA 16S. As análises dos índices de riqueza (Chao 1), diversidade (Shannon e Simpson) e beta diversidade indicaram que a comunidade bacteriana do colostro de pluríparas e primíparas não diferiram entre a categoria animal (teste t, p < 0,05). Os filos Firmicutes, Tenericutes, Kiritimatiellaeota e Fibrobacteres foram significativamente mais abundantes (p < 0,05) em pluríparas quando comparado às primíparas, enquanto que Proteobacteria, Actinobacteria, Cloacimonetes e Fusobacteria foram mais abundantes (White’s nonparametric t-test, p < 0,05) em primíparas quando comparados com as pluríparas. As famílias Lachnospiraceae, Ruminococcaceae, Prevotellaceae, Rikenellaceae e Christensenellaceae foram mais abundantes nas pluríparas (White’s non-parametric t-test, p < 0,05) quando comparada às primíparas. Enquanto que Weeksellaceae e Burkholderiaceae foram mais abundantes nas primíparas quando comparado às pluríparas (White’s non- parametric t-test, p < 0,05). Os gêneros mais abundantes (>0,1%) foram Pseudomonas (1,2 e 0,64%), Fibrobacter (0,26 e 0,47%), Stenotrophomonas (0,53 e 0,21%), Sphingobacterium (0,48 e 0,16%) e Brevundimonas (0,34 e 0,12%), nas primíparas e pluríparas, respectivamente. As OTUs que foram identificadas em pelo menos 50% dos animais de uma das categorias (n ≥ 4) foram selecionadas para a análise de espécies indicadoras, resultando no total de 368 OTUs. As OTUs Otu01030 e Otu00891 foram classificadas como membros das famílias Rikenellaceae e Christensenellaceae, enquanto que a Otu00721 foi classificada como pertencente ao gênero Butyrivibrio e Otu00094 como do gênero Prevotella. Essas OTUs foram identificadas como indicadoras nas amostras de colostro de vacas pluríparas, independentemente se foram ou não observadas nas amostras de primíparas. A Otu00101, identificada como Brevundimonas, foi considerada como indicadora das amostras de colostro de primíparas, sendo observada apenas nas amostras desses animais. A análise da betadiversidade das fezes indicou que bezerros de pluríparas e bezerros de primíparas foram significativamente semelhantes entre si, assim como as pluríparas e primíparas (ANOSIM, p < 0,05). Os índices de riqueza e diversidade (Chao 1, Shannon e Simpson) indicaram que a comunidade bacteriana das fezes de pluríparas e primíparas, assim como das fezes dos bezerros de pluríparas e primíparas não diferem significativamente entre si (teste t, p > 0,05). Foram identificadas 47 famílias compartilhadas nas fezes dos animais adultos e bezerros após o nascimento, sendo que Ruminococcaceae, Lachnospiraceae e Christensenellaceae foram mais abundantes nas pluríparas e primíparas, enquanto Enterobacteriaceae, Clostridiaceae_1 e Ruminococcaceae foram mais abundantes nos bezerros. Apenas a família Lactobacillaceae foi mais abundante em bezerros nascidos de primíparas quando comparado aos bezerros nascidos de pluríparas (White’s non-parametric ttest, p < 0,05). O gênero Ruminococcus_1 foi significativamente mais abundante (White’s non-parametric t-test, p < 0,05) nas primíparas (0,74 ± 0,22%) quando comparado às pluríparas (0,23 ± 0,16%), enquanto Lactobacillus e Faecalibacterium apresentaram abundância relativa significativamente maior (White’s non-parametric t-test, p < 0,05) nas fezes dos bezerros de primíparas quando comparado à microbiota fecal dos bezerros de multíparas. Dessa forma, o colostro de pluríparas e primíparas diferem na abundância relativa dos filotipos e Lachnospiraceae, Ruminococcaceae e Prevotellaceae foram os grupos mais abundantes no colostro de multíparas. As fezes de bezerros originados de primíparas possuem microbiota com maior abundância de Lactobacillus e Faecalibacterium, os quais parecem estar relacionados com a saúde e desempenho dos bezerros.
In this work, the objective was to evaluate the composition of the microbiota of the colostrum and the feces of multiparous and primiparous cows, as well as the microbiota of the feces of calves in the postpartum period. Analyzes of the noncultured colostrum microbiota (1, 2 and 3 days postpartum) and the fecal microbiota of cows and deep sequencing of the 16S rRNA gene. Analyzes of the species richness indices (Chao 1), diversity (Shannon and Simpson) and beta diversity indicated that the bacterial colostrum community of multiparous and primiparous did not differ between the animal category (t-test, p < 0.05). Phyla Firmicutes, Tenericutes, Kiritimatiellaeota and Fibrobacteres were significantly more abundant (p < 0.05) in multiparous than primiparous, while Proteobacteria, Actinobacteria, Cloacimonetes and Fusobacteria were more abundant (White's non-parametric t- test, p < 0.05) in primiparous when compared to multiparous. The families Lachnospiraceae, Ruminococcaceae, Prevotellaceae, Rikenellaceae and Christensenellaceae were more abundant in pluriparous cows (White's nonparametric t-test, p < 0.05) when compared to the primiparous. The families Weeksellaceae and Burkholderiaceae were more abundant in primiparous compared to multiparous cows (White's non-parametric t-test, p < 0.05). The most abundant genera (> 0.1%) were Pseudomonas (1.2 and 0.64%), Fibrobacter (0.26 and 0.47%), Stenotrophomonas (0.53 and 0.21%), Sphingobacterium (0.48 and 0.16%) and Brevundimonas (0.34 and 0.12%) in the primiparous and multiparous cows, respectively. OTUs that were identified in at least 50% of the animals from one of the categories (n ≥ 4) were selected for the analysis of indicator species, resulting in 368 OTUs. OTU01030 and OTU00891 were classified as members of the families Rikenellaceae and Christensenellaceae, while OTU00721 was classified as belonging to the genus Butyrivibrio and Otu00094 as from the genus Prevotella. These OTUs were identified as indicators in colostrum samples from pluriparous cows, regardless if they were or not observed in samples from primiparous animals. OTU00101, identified as Brevundimonas, was considered as an indicator of the primiparous colostrum samples, being observed only in colostrum from these animals. Analysis of fecal beta diversity indicated that multiparous and primiparous calves were significantly similar to each other, as well as multiparous and primiparous (ANOSIM, p < 0.05). The richness and diversity indices (Chao 1, Shannon and Simpson) indicated that the bacterial community in the feces of multiparous and primiparous animals, as well as in feces of calves from multiparous and primiparous, did not differ significantly (test t, p > 0.05). A total of 47 bacterial families were found in the feces of adult animals and calves after birth. Ruminococcaceae, Lachnospiraceae and Christensenellaceae were more abundant in the multiparous and primiparous animals, whereas Enterobacteriaceae, Clostridiaceae and Ruminococcaceae were more abundant in calves. Only the Lactobacillaceae family was more abundant in primiparous calves when compared to multiparous calves (White’s non-parametric t-test , p < 0.05). The genus Ruminococcus_1 was significantly more abundant (White’s non-parametric t-test, p < 0.05) in primiparous animals (0.74 ± 0.22%) when compared to the multiparous cows (0.23 ± 0.16%), while Lactobacillus and Faecalibacterium showed greater relative abundance (White’s non-parametric t-test, p < 0.05) in feces of primiparous calves when compared to the fecal microbiota of multiparous calves. Therefore, the colostrum of multiparous and primiparous differs in relative abundance of the phylotypes and the colostrum of multiparous cows is more diverse than the primiparous in the second day after calving. Lachnospiraceae, Ruminococcaceae and Prevotellaceae were the most abundant groups in the colostrum from multiparous cows. The feces of calves originated from primiparous animals harbor a microbiota with greater abundance of Lactobacillus and Faecalibacterium, which appears to be related with health and performance of the calves.
In this work, the objective was to evaluate the composition of the microbiota of the colostrum and the feces of multiparous and primiparous cows, as well as the microbiota of the feces of calves in the postpartum period. Analyzes of the noncultured colostrum microbiota (1, 2 and 3 days postpartum) and the fecal microbiota of cows and deep sequencing of the 16S rRNA gene. Analyzes of the species richness indices (Chao 1), diversity (Shannon and Simpson) and beta diversity indicated that the bacterial colostrum community of multiparous and primiparous did not differ between the animal category (t-test, p < 0.05). Phyla Firmicutes, Tenericutes, Kiritimatiellaeota and Fibrobacteres were significantly more abundant (p < 0.05) in multiparous than primiparous, while Proteobacteria, Actinobacteria, Cloacimonetes and Fusobacteria were more abundant (White's non-parametric t- test, p < 0.05) in primiparous when compared to multiparous. The families Lachnospiraceae, Ruminococcaceae, Prevotellaceae, Rikenellaceae and Christensenellaceae were more abundant in pluriparous cows (White's nonparametric t-test, p < 0.05) when compared to the primiparous. The families Weeksellaceae and Burkholderiaceae were more abundant in primiparous compared to multiparous cows (White's non-parametric t-test, p < 0.05). The most abundant genera (> 0.1%) were Pseudomonas (1.2 and 0.64%), Fibrobacter (0.26 and 0.47%), Stenotrophomonas (0.53 and 0.21%), Sphingobacterium (0.48 and 0.16%) and Brevundimonas (0.34 and 0.12%) in the primiparous and multiparous cows, respectively. OTUs that were identified in at least 50% of the animals from one of the categories (n ≥ 4) were selected for the analysis of indicator species, resulting in 368 OTUs. OTU01030 and OTU00891 were classified as members of the families Rikenellaceae and Christensenellaceae, while OTU00721 was classified as belonging to the genus Butyrivibrio and Otu00094 as from the genus Prevotella. These OTUs were identified as indicators in colostrum samples from pluriparous cows, regardless if they were or not observed in samples from primiparous animals. OTU00101, identified as Brevundimonas, was considered as an indicator of the primiparous colostrum samples, being observed only in colostrum from these animals. Analysis of fecal beta diversity indicated that multiparous and primiparous calves were significantly similar to each other, as well as multiparous and primiparous (ANOSIM, p < 0.05). The richness and diversity indices (Chao 1, Shannon and Simpson) indicated that the bacterial community in the feces of multiparous and primiparous animals, as well as in feces of calves from multiparous and primiparous, did not differ significantly (test t, p > 0.05). A total of 47 bacterial families were found in the feces of adult animals and calves after birth. Ruminococcaceae, Lachnospiraceae and Christensenellaceae were more abundant in the multiparous and primiparous animals, whereas Enterobacteriaceae, Clostridiaceae and Ruminococcaceae were more abundant in calves. Only the Lactobacillaceae family was more abundant in primiparous calves when compared to multiparous calves (White’s non-parametric t-test , p < 0.05). The genus Ruminococcus_1 was significantly more abundant (White’s non-parametric t-test, p < 0.05) in primiparous animals (0.74 ± 0.22%) when compared to the multiparous cows (0.23 ± 0.16%), while Lactobacillus and Faecalibacterium showed greater relative abundance (White’s non-parametric t-test, p < 0.05) in feces of primiparous calves when compared to the fecal microbiota of multiparous calves. Therefore, the colostrum of multiparous and primiparous differs in relative abundance of the phylotypes and the colostrum of multiparous cows is more diverse than the primiparous in the second day after calving. Lachnospiraceae, Ruminococcaceae and Prevotellaceae were the most abundant groups in the colostrum from multiparous cows. The feces of calves originated from primiparous animals harbor a microbiota with greater abundance of Lactobacillus and Faecalibacterium, which appears to be related with health and performance of the calves.
Descrição
Palavras-chave
Microbiologia agrícola, Colostro, Bovinos, Microbiota, Sequenciamento de nucleotídeo
Citação
SILVA, Juliana Soares da. Análise da microbiota do colostro de vacas pluríparas e primíparas e de fezes de vacas e bezerros leiteiros no pós parto. 2018. 88 f. Tese (Doutorado em Microbiologia Agrícola) - Universidade Federal de Viçosa, Viçosa. 2018.