Abstract
A recent study reported the existence of a diverse microbiota in 5-to-7-month-old calf fetuses, suggesting that colonization of the bovine gut with so-called “pioneer” microbiota may begin during mid-gestation. In the present study, we investigated the microbiota in bovine fetuses at early gestation. Amniotic and allantoic fluids, and intestinal and placental (cotyledon) tissue samples harvested from fetuses (n = 33) on day 83 of gestation were processed for the assessment of fetal microbiota using 16S rRNA gene sequencing. The sequencing results revealed that a diverse and complex microbial community was present in allantoic and amniotic fluids, and fetal intestine and placenta on day 83 of gestation in beef cattle. Microbial community structure was significantly different between allantoic and amniotic fluid, and intestinal and placental microbiota (0.047 ≥ R(2) ≥ 0.019, P ≤ 0.031). Allantoic fluid had a greater (P < 0.05) microbial richness (number of OTUs) (122 ± 10) compared to amniotic fluid (84 ± 6), intestine (63 ± 7) and placenta (66 ± 6). Microbial diversity (Shannon index) was similar for the intestinal and placental samples, and both were less diverse compared to the fetal fluid microbiota (P < 0.05). At the phylum level, 39 different archaeal and bacterial phyla were detected across all fetal samples, with Proteobacteria (55%), Firmicutes (16.2%), Actinobacteria (13.6%) and Bacteroidetes (5%) predominating. Among the 20 most relatively abundant bacterial genera, Acidovorax, Acinetobacter, Brucella, Corynebacterium, Enterococcus, Exiguobacterium and Stenotrophomonas differed by fetal sample type (P < 0.05). A total of 55 taxa were shared among the four different microbial communities. qPCR of bacteria in the intestine and placenta samples as well as scanning electron microscopy imaging of fetal fluids provided additional evidence for the presence of a microbiota in these samples. Overall, the results of this study indicate that colonization with pioneer microbiota may occur during early gestation in bovine fetuses.