Uterine and vaginal bacterial community diversity prior to artificial insemination between pregnant and nonpregnant postpartum cows1.

The present study evaluated the bovine vaginal and uterine bacterial community diversity and its relationship to fertility. Postpartum beef cows (n = 68) were synchronized beginning on day -21 and ending with timed artificial insemination (TAI) on day 0. Pregnancy was diagnosed 30 d after TAI. Uterine and vaginal flushes were collected on day -21, -9, and -2 for bacterial DNA extraction to sequence the V1 to V3 hypervariable regions of the 16S rRNA gene. Results indicated a decrease in the number of bacterial species over time in the uterus of resulting pregnant and nonpregnant beef cows (P < 0.0001). Principal coordinate analyses (PCoA) depicted clustering of samples, indicating closely related bacterial communities, by day in the uterus and vagina (P < 0.0001). At day -2, uterine samples from nonpregnant and pregnant animals clustered separately (P < 0.0001), with nonpregnant animal samples clustering tightly together. Overall, the current study suggests the shift in the reproductive bacterial communities' diversity and phylogenetic relationship leading up to the time of breeding may contribute to successful pregnancy establishment.

Bacterial taxonomic composition of the postpartum cow uterus and vagina prior to artificial insemination1.

The current study characterized the taxonomic composition of the uterine and vaginal bacterial communities during estrous synchronization up to timed artificial insemination (TAI). Postpartum beef cows (n = 68) were subjected to pre-synchronization step 21 d prior to TAI (day -21), followed by an industry standard 7 Day Co-Synch on day -9 and TAI on day 0. Uterine and vaginal flushes were collected on days -21, -9, and -2 of the protocol and pH was immediately recorded. Pregnancy was determined by transrectal ultrasound on day 30. Bacterial DNA was extracted and sequenced targeting the V1 to V3 hypervariable regions of the 16S rRNA bacterial gene. Results indicated 34 different phyla including 792 different genera present between the uterus and vagina. Many differences in the relative abundance of bacterial phyla and genera occurred between resulting pregnancy statuses and among protocol days (P < 0.05). At day -2, multiple genera were present in >1% abundance of nonpregnant cows but <1% abundance in pregnant cows (P < 0.05). Uterine pH increased in nonpregnant cows but decreased in pregnant cows (P > 0.05). Overall, our study indicates bacterial phyla and genera abundances shift over time and may potentially affect fertility by altering the reproductive tract environment.

Effect of complexed trace minerals on cumulus-oocyte complex recovery and in vitro embryo production in beef cattle1,2.

The objective of this experiment was to evaluate the impact of complexed trace mineral supplementation on ovum pick-up (OPU) and in vitro embryo production in lactating beef cows. Thirty days prior to fixed-time artificial insemination (FTAI; day -30), 68 postpartum cows were stratified by BW, BCS, and parity before being randomly assigned to 10 pens of either a treatment (TRT; n = 5) or a control (CNT; n = 5) group. Each group received a weekly mineral supplement allotment of 1.16 kg × week-1 × cow-calf pair-1 for 14 wk. Cows assigned to the TRT group received a mineral supplement that contained amino acid complexes of zinc, copper, and manganese, as well as cobalt glucoheptonate (Availa Plus; Zinpro Corp., Eden Prairie, MN, USA), while cows assigned to the CNT group received a mineral supplement that was formulated to contain similar concentrations of these trace minerals from inorganic sources. All cows were submitted to a 7 d CO-Synch + CIDR protocol on day -10 and bred using FTAI on day 0. Pregnancy diagnosis was performed on day 28 and nonpregnant cows were removed. All pregnant cows were subjected to ovum pick-up (OPU) on day 52 and 67 of gestation. Cumulus-oocyte complexes (COCs) were evaluated and graded prior to in vitro fertilization (IVF). Analysis of variance was conducted to determine effects of treatment on response variables, and pen was considered the experimental unit. Supplement consumption did not differ (P = 0.48) between treatments (1.16 ± 0.12 vs. 1.07 ± 0.15 kg of DM × week-1 × cow-calf pair-1 for TRT and CNT, respectively). Total COC recovery was greater (P = 0.03) from TRT when compared with CNT cows (22.4 ± 2.0 vs. 16.4 ± 1.4 COCs × pen-1, respectively) and the number of COCs meeting maturation criteria was increased in TRT cows (P = 0.05) when compared with CNT cows (15.9 ± 1.6 vs. 11.8 ± 1.0 COCs × pen-1, respectively). Production of transferable embryos tended to be greater (P = 0.06) for TRT than CNT cows (4.7 ± 0.6 vs. 2.7 ± 0.7 embryos × pen-1, respectively). Furthermore, when expressed as a ratio, the number of recovered COCs meeting maturation criteria that were required to produce a transferable embryo tended to be lower for TRT than CNT cows (3.10 ± 0.93 vs. 7.02 ± 1.60; P = 0.06). In summary, complete replacement with complexed trace mineral improved COC recovery and in vitro embryo production when compared with inorganic forms of these trace minerals in beef cows.

Vaginal and Uterine Bacterial Communities in Postpartum Lactating Cows.

Reproductive inefficiency in cattle has major impacts on overall productivity of cattle operations, increasing cost of production, and impacting the sustainability of the cattle enterprise. Decreased reproductive success and associated disease states have been correlated with the presence of specific microbes and microbial community profiles, yet details of the relationship between microbial communities and host physiology are not well known. The present study profiles and compares the microbial communities in the bovine uterus and vagina using 16S rRNA sequencing of the V1-V3 hypervariable region at the time of artificial insemination. Significant differences (p < 0.05) between the vaginal and uterine communities were observed at the level of α-diversity metrics, including Chao1, Shannon's Diversity Index, and observed OTU. Greater clustering of vaginal OTU was apparent in principal coordinate analysis compared to uterine OTU, despite greater diversity in the vaginal community in both weighted and unweighted UniFrac distance matrices (p < 0.05). There was a significantly greater relative abundance of unassigned taxa in the uterus (p = 0.008), otherwise there were few differences between the overall community profiles. Both vaginal and uterine communities were dominated by Firmicutes, although the relative abundance of rRNA sequences corresponding to species in this phylum was significantly (p = 0.007) lower in the uterine community. Additional differences were observed at the genus level, specifically in abundances within Clostridium (p = 0.009), Anaerofustis (p = 0.018), Atopobium (p = 0.035), Oscillospira (p = 0.035), 5-7N15 (p = 0.035), Mycoplasma (p = 0.035), Odoribacter (p = 0.042), and within the families Clostridiaceae (p = 0.006), Alcaligenaceae (p = 0.021), and Ruminococcaceae (p = 0.021). Overall, the comparison revealed differences and commonalities among bovine reproductive organs, which may be influenced by host physiology. The increased abundance of unassigned taxa found in the uterus may play a significant biological role in the reproductive status of the animal. The study represents an initial dataset for comparing bacterial communities prior to establishment of pregnancy.