244 Fertility Outcomes Associated with Reproductive Tract Cytokines and Microbiota in Postpartum Beef Cows

Abstract Local immune activity in the reproductive tract is crucial in the response to uterine diseases, normal reproductive functions, and establishing pregnancy. Few studies have evaluated the influence of the local immune environment of the reproductive tract on fertility outcomes. The objectives were to 1) evaluate reproductive cytokine concentrations in postpartum cows undergoing estrus synchronization followed by timed artificial insemination (TAI) and 2) correlate reproductive bacterial communities with cytokine concentrations. Angus cows (n = 20) were subjected to a 7-Day Co-Synch protocol with pre-synchronization beginning 21 days prior (d -21) to TAI (d 0). Uterine and vaginal flushes were collected on d -21 and -2. Pregnancy was determined by transrectal ultrasound on d 30. Bacterial community profiling and analyses were conducted targeting the V1 to V3 hypervariable regions of the 16S rRNA gene. Cytokine concentrations for interleukin (IL)-1b, IL-6, IL-10, and transforming growth factor beta (TGF-β) were determined by commercial ELISA kits. Concentration data were analyzed using PROC MIXED and correlations using Pearson correlation in SAS. No differences were detected in vaginal samples (P >0.05). No differences in IL-10 or IL-1b concentrations were detected in uterine samples (P >0.05). Uterine TGF-β concentrations were greater in resulting pregnant than non-pregnant cows (44.0 ± 13.4 pg/mL vs 14.7 ± 4.9 pg/mL; P = 0.05). Uterine TGF-β was negatively correlated with the relative abundance of genera Treponema (r = -0.668; P = 0.05) in resulting non-pregnant cows on d -21. Uterine IL-6 concentrations were greater in resulting non-pregnant than pregnant cows (198.7 ± 21.8 pg/mL vs 144.3 ± 16.1 pg/mL; P = 0.05). Uterine IL-6 and the relative abundance of genera Butyrivibrio were positively correlated (r = 0.742; P = 0.02) in resulting non-pregnant cows on d -21. These results suggest possible relationships between uterine bacterial communities and cytokines prior to TAI that may ultimately affect fertility outcomes in beef cattle.

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181 Uterine and vaginal bacteria and cytokine profiles prior to artificial insemination between resulting pregnant and open postpartum beef cows

Journal of Animal Science ◽

10.1093/jas/skz397.099 ◽

2020 ◽

Vol 98 (Supplement_2) ◽

pp. 43-44

Author(s):

Rebecca Poole ◽

Taylor B Ault ◽

Phillip R Myer ◽

Andrea Lear ◽

Ky Pohler

Keyword(s):

Beef Cattle ◽

Relative Abundance ◽

Bacterial Communities ◽

Reproductive Tract ◽

Pearson Correlation ◽

Beef Cows ◽

Rrna Gene ◽

Concentration Data ◽

Positive Correlation ◽

Cytokine Profiles

Abstract Numerous factors impact reproductive success in beef cattle; however, the presence and activity of bacteria in the reproductive tract and its effects on fertility is relatively unknown. The objective of this study was to evaluate the relationship between reproductive tract bacterial communities and cytokine profiles prior to timed-AI (TAI) and compare between resulting pregnancy status at d30. Sixty-eight beef cows, 80 ± 2.6 d postpartum at TAI, were synchronized and subjected to TAI on d0. Pregnancy was diagnosed via transrectal ultrasonography on d30. Uterine and vaginal flushes were collected on d -21 and -2 for bacterial DNA extraction to sequence the V1-V3 hypervariable regions of the 16S rRNA gene and to measure pro-inflammatory [interlukin-6 (IL-6)] and anti-inflammatory [IL-10 and transforming growth factor-beta (TGFβ)] cytokine concentrations using validated commercial ELISAs. Concentration data were analyzed using repeated measures in PROC MIXED of SAS and correlations were performed in SAS using Pearson correlation. There were no differences in cytokine concentrations by day or status x day. There were no differences in uterine IL-6 or IL-10 concentrations (P > 0.05). Uterine TGFβ concentrations were overall elevated in pregnant cows compared to open cows (72.6 vs. 13.7 ± 12.1 pg/mL, respectively; P < 0.05). On d -2, a positive correlation exists between TGFβ and the relative abundance of Actinobacteria in the uterus of pregnant cows (r=0.93, P = 0.02). There were no differences in vaginal IL-6 or TGFβ concentrations (P > 0.05). On d -2, vaginal IL-10 concentrations were elevated in open cows compared to pregnant cows (741.6 vs. 602.1 ± 47.0 pg/mL, respectively; P < 0.05) and a positive correlation exists between IL-10 and the relative abundance of Lentisphaerae in the vagina of pregnant cows (r=0.89, P = 0.04). These data suggest a possible relationship between bacterial communities and cytokines concentrations within the reproductive tract of beef cattle which may affect fertility.

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Evaluation of Reproductive Tract Cytokines in Post-partum Beef Cows Relating to Reproductive Microbiota and Fertility Outcomes

Frontiers in Animal Science ◽

10.3389/fanim.2021.704714 ◽

2021 ◽

Vol 2 ◽

Author(s):

Rebecca K. Poole ◽

Taylor B. Ault-Seay ◽

Rebecca R. Payton ◽

Phillip R. Myer ◽

Andrea S. Lear ◽

...

Keyword(s):

Relative Abundance ◽

Transforming Growth Factor Beta ◽

Bacterial Communities ◽

Reproductive Tract ◽

Transforming Growth Factor ◽

Post Partum ◽

Transrectal Ultrasound ◽

Beef Cows ◽

Pregnancy Status ◽

Uterine Diseases

The activity of the immune system in the reproductive tract has been proven to be crucial in the response to uterine diseases, normal reproductive functions, and tolerance to the allogeneic fetus during pregnancy. The objectives of the current study were to (1) evaluate uterine and vaginal cytokine concentrations in postpartum cows undergoing estrus synchronization followed by timed artificial insemination (TAI) and (2) correlate bacterial communities with cytokine concentrations. Postpartum Angus cows (n = 20) were subjected to a 7-Day Co-Synch protocol with pre-synchronization beginning 21 days prior (d −21) to TAI (d 0). Uterine and vaginal flushes were collected on d −21 and −2. Pregnancy was determined by transrectal ultrasound on d 30. Cytokines include interleukin (IL)-1b, IL-6, IL-10, transforming growth factor beta (TGF-β), and immunoglobin A (IgA) and concentrations were determined by commercial ELISA kits. No differences by day or pregnancy status in cytokine concentrations were detected in vaginal samples. No differences by day or pregnancy status in IgA, IL-10, or IL-1b concentrations were detected in uterine samples. Overall TGF-β concentrations in the uterus were greater in resulting pregnant than non-pregnant cows (44.0 ± 13.4 pg/mL vs. 14.7 ± 4.9 pg/mL; P = 0.047). Uterine TGF-β was correlated with the relative abundance of genera Treponema (r = −0.668; P = 0.049) in resulting non-pregnant cows on d −21 and with the relative abundance of genera Ureaplasma (r = 0.901; P = 0.0004) in resulting pregnant cows on d −2. In resulting pregnant animals, a tendency for a strong correlation was detected between d −2 progesterone concentrations and uterine TGF-β concentrations (r = 0.591, P = 0.07). Overall IL-6 concentrations in the uterus were greater in resulting non-pregnant than pregnant cows (198.7 ± 21.8 pg/mL vs. 144.3 ± 16.1 pg/mL; P = 0.045). A correlation was also detected between uterine IL-6 concentrations and the relative abundance of genera Butyrivibrio (r = 0.742; P = 0.022) in resulting non-pregnant cows on d −21. These results suggest possible relationships between different bacterial communities and cytokine concentrations within the uterus of beef cattle prior to TAI that may ultimately affect fertility outcomes.

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87 Bacterial communities of the uterus and vagina between resulting pregnant and non-pregnant postpartum beef cows

Journal of Animal Science ◽

10.1093/jas/skz122.089 ◽

2019 ◽

Vol 97 (Supplement_2) ◽

pp. 49-49

Author(s):

Taylor B Ault ◽

Brooke A Clemmons ◽

Sydney T Reese ◽

Gessica A Franco ◽

Felipe G Dantas ◽

...

Keyword(s):

Bacterial Communities ◽

Reproductive Tract ◽

Bacterial Species ◽

Transrectal Ultrasound ◽

Beef Cows ◽

Rrna Gene ◽

Hypervariable Regions ◽

Pregnancy Status ◽

Uterine Environment ◽

Bacterial Dna Extraction

Abstract The prevalence of reproductive losses costing 1 billion dollars annually to the beef and dairy industries has led researchers to evaluate strategies to improve reproductive efficiency. The microbiome of the reproductive tract has been a recent focus in humans due to its effects on health and fertility; however, little research has evaluated the uterine and vaginal microbiomes of cattle. The present study evaluated the reproductive tract bacterial communities of postpartum beef cows undergoing estrus synchronization and the differences between cows who became pregnant and those who failed to conceive. Forty postpartum Angus cows were subjected to a 7 Day Co-Synch protocol with a pre-synchronization step 21 days prior (day -21) to artificial insemination (AI). Uterine and vaginal flushes were collected on day -21, -9, and -2 of the protocol for pH and bacterial DNA extraction to sequence the V1-V3 hypervariable regions of the 16S rRNA gene. Transrectal ultrasound was performed 30 days after AI for pregnancy diagnosis. Ten pregnant and non-pregnant animals were selected and used for analysis. Decreased numbers of bacterial species present in the uterus of cows were identified, independent of pregnancy status, leading up to AI with an average of 1180 ± 97 species on day -21 to 427 ± 99 on day -2 (P < 0.0001). Principal coordinate analyses depicted significant clustering of bacterial communities by day in the uterus (P = 0.001) and vagina (P = 0.001) as well as pregnancy status in the uterus at day -2 (P = 0.005). Additionally, at day -2, multiple significantly different uterine bacterial genera were present at >1% in non-pregnant cows while present at <1% in pregnant cows. Uterine pH increased in non-pregnant cows but decreased in pregnant cows. In summary, the results suggest uterine bacterial abundances prior to AI may influence the uterine environment and reproductive outcomes.

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Environmental Factors Shape the Epiphytic Bacterial Communities of Gracilariopsis Lemaneiformis

10.21203/rs.3.rs-144624/v1 ◽

2021 ◽

Author(s):

Pengbing Pei ◽

Hong Du ◽

Muhammad Aslam ◽

Honghao Liang ◽

Hui Wang ◽

...

Keyword(s):

Environmental Factors ◽

Relative Abundance ◽

Bacterial Communities ◽

High Throughput Sequencing ◽

Inorganic Nitrogen ◽

Pearson Correlation ◽

Critical Role ◽

Gracilariopsis Lemaneiformis ◽

Rrna Gene ◽

Epiphytic Bacteria

Abstract Macroalgae host variety of symbionts on their surface which play critical role in their growth and development processes. Nevertheless, the complete understanding of this interaction of epiphytic bacteria and host algae is still in its infancy. This study comprehensively analyses epiphytic bacterial communities composition of red macroalga Gracilariopsis lemaneiformis and environmental factors such as nitrogen and phosphorus which shape the composition of epiphytic bacterial communities of G. lemaneiformis and variation of epiphytic bacterial communities composition at different geographical location. The composition and structure of epiphytic bacterial communities were characterized using high throughput sequencing data of the V3-V4 hypervariable region of 16S rRNA gene amplicon sequencing. The epiphytic bacterial communities composition data revealed that epiphytic bacteria varied significantly among three different geographic locations i) Nan’ao Island (NA) (ii) Lianjiang County (LJ) and iii) Nanri Island (NR) in China. Redundancy analysis (RDA) showed that the relative abundance of Bacteroidetes, Firmicutes, Verrucomicrobia and Epsilonbacteraeota at NR were strongly positively correlated with total nitrogen (TN), total phosphorus (TP), nitrate nitrogen (NO3-N), and dissolved inorganic nitrogen (DIN), whereas negatively correlated with nitrite nitrogen (NO2-N). In addition, the relative abundance of cyanobacteria at NA and LJ were strongly positively correlated with NO2-N, whereas negatively correlated with TN, TP, NO3-N, and DIN. Furthermore, the results of Mantel test indicated that the epiphytic bacterial communities composition is significantly correlated with these environmental factors, which are also proved by Pearson correlation analysis. In conclusion, it is proposed that environmental factors such as NO3-N and DIN play key role in the communities composition of epiphytic bacteria in G. lemaneiformis. Our study provides important baseline knowledge for the communities composition of epiphytic bacteria in G. lemaneiformis and their correlation among themselves as well as with their surrounding environmental factors.

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A Grain-Based SARA Challenge Affects the Composition of Epimural and Mucosa-Associated Bacterial Communities throughout the Digestive Tract of Dairy Cows

Animals ◽

10.3390/ani11061658 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1658

Author(s):

Jan C. Plaizier ◽

Anne-Mette Danscher ◽

Paula A. Azevedo ◽

Hooman Derakhshani ◽

Pia H. Andersen ◽

...

Keyword(s):

Digestive Tract ◽

Relative Abundance ◽

Bacterial Communities ◽

Dry Matter ◽

Control Diet ◽

Rrna Gene ◽

Ruminal Acidosis ◽

Clustering Patterns ◽

The 16S Rrna Gene ◽

Distal Jejunum

The effects of a subacute ruminal acidosis (SARA) challenge on the composition of epimural and mucosa-associated bacterial communities throughout the digestive tract were determined in eight non-lactating Holstein cows. Treatments included feeding a control diet containing 19.6% dry matter (DM) starch and a SARA-challenge diet containing 33.3% DM starch for two days after a 4-day grain step-up. Subsequently, epithelial samples from the rumen and mucosa samples from the duodenum, proximal, middle and distal jejunum, ileum, cecum and colon were collected. Extracted DNA from these samples were analyzed using MiSeq Illumina sequencing of the V4 region of the 16S rRNA gene. Distinct clustering patterns for each diet existed for all sites. The SARA challenge decreased microbial diversity at all sites, with the exception of the middle jejunum. The SARA challenge also affected the relative abundances of several major phyla and genera at all sites but the magnitude of these effects differed among sites. In the rumen and colon, the largest effects were an increase in the relative abundance of Firmicutes and a reduction of Bacteroidetes. In the small intestine, the largest effect was an increase in the relative abundance of Actinobacteria. The grain-based SARA challenge conducted in this study did not only affect the composition and cause dysbiosis of epimural microbiota in the rumen, it also affected the mucosa-associated microbiota in the intestines. To assess the extent of this dysbiosis, its effects on the functionality of these microbiota must be determined in future.

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Endophytic bacterial communities of oilseed rape associate with genotype-specific resistance against Verticillium longisporum

FEMS Microbiology Ecology ◽

10.1093/femsec/fiz188 ◽

2019 ◽

Vol 96 (1) ◽

Cited By ~ 1

Author(s):

Stefanie P Glaeser ◽

Iulian Gabur ◽

Hossein Haghighi ◽

Jens-Ole Bartz ◽

Peter Kämpfer ◽

...

Keyword(s):

Oilseed Rape ◽

Relative Abundance ◽

Bacterial Communities ◽

Quantitative Resistance ◽

Bacterial Community Composition ◽

Phylogenetic Group ◽

Rrna Gene ◽

Host Genotype ◽

Brassica Napus L ◽

Verticillium Longisporum

ABSTRACT Associations of endophytic bacterial community composition of oilseed rape (Brassica napus L.) with quantitative resistance against the soil-borne fungal pathogen Verticillium longisporum was assessed by 16S rRNA gene amplicon sequencing in roots and hypocotyls of four plant lines with contrasting genetic composition in regard to quantitative resistance reactions. The plant compartment was found to be the dominating driving factor for the specificity of bacterial communities in healthy plants. Furthermore, V. longisporum infection triggered a stabilization of phylogenetic group abundance in replicated samples suggesting a host genotype-specific selection. Genotype-specific associations with bacterial phylogenetic group abundance were identified by comparison of plant genotype groups (resistant versus susceptible) and treatment groups (healthy versus V. longisporum-infected) allowing dissection into constitutive and induced directional association patterns. Relative abundance of Flavobacteria, Pseudomonas, Rhizobium and Cellvibrio was associated with resistance/susceptibility. Relative abundance of Flavobacteria and Cellvibrio was increased in resistant genotypes according to their known ecological functions. In contrast, a higher relative abundance of Pseudomonas and Rhizobium, which are known to harbor many species with antagonistic properties to fungal pathogens, was found to be associated with susceptibility, indicating that these groups do not play a major role in genetically controlled resistance of oilseed rape against V. longisporum.

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Semen Modulates Inflammation and Angiogenesis in the Reproductive Tract of Female Rabbits

Animals ◽

10.3390/ani10122207 ◽

2020 ◽

Vol 10 (12) ◽

pp. 2207

Author(s):

Jaume Gardela ◽

Amaia Jauregi-Miguel ◽

Cristina A. Martinez ◽

Heriberto Rodríguez-Martinez ◽

Manel López-Béjar ◽

...

Keyword(s):

Gene Expression ◽

Growth Factor ◽

Transforming Growth Factor Beta ◽

Reproductive Tract ◽

Transforming Growth Factor ◽

Interleukin 10 ◽

Breeding Systems ◽

Preimplantation Embryos ◽

Natural Mating ◽

Anti Inflammatory

The maternal environment modulates immune responses to facilitate embryo development and ensure pregnancy. Unraveling this modulation could improve the livestock breeding systems. Here it is hypothesized that the exposure of the female rabbit reproductive tract to semen, as well as to early embryos, modulates inflammation and angiogenesis among different tissue segments. qPCR analysis of the gene expression changes of the anti-inflammatory interleukin-10 (IL10) and transforming growth factor beta family (TGFβ1–3) and the angiogenesis mediator vascular endothelial growth factor (VEGF-A) were examined in response to mating or insemination with sperm-free seminal plasma (SP). Reproductive tract segment (cervix to infundibulum) samples were obtained in Experiment 1, 20 h after gonadotropin-releasing hormone (GnRH) stimulation (control), natural mating (NM) or vaginal infusion with sperm-free SP (SP-AI). Additionally, segmented samples were also obtained at 10, 24, 36, 68 or 72 h after GnRH-stimulation and natural mating (Experiment 2). The results of gene expression, analyzed by quantitative PCR, showed that NM effects were mainly localized in the uterine tissues, depicting clear temporal variation, while SP-AI effects were restricted to the oviduct. Changes in anti-inflammatory and angiogenesis mediators indicate an early response in the uterus and a late modulation in the oviduct either induced by semen or preimplantation embryos. This knowledge could be used in the implementation of physiological strategies in breeding systems to face the new challenges on rabbit productivity and sustainability.

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Changes of Bacterial Communities in Response to Prolonged Hydrodynamic Disturbances in the Eutrophic Water-Sediment Systems

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph16203868 ◽

2019 ◽

Vol 16 (20) ◽

pp. 3868

Author(s):

Haomiao Cheng ◽

Ling Cheng ◽

Liang Wang ◽

Tengyi Zhu ◽

Wei Cai ◽

...

Keyword(s):

Relative Abundance ◽

Bacterial Communities ◽

16S Rrna Gene Sequencing ◽

Aquatic Environments ◽

Rrna Gene ◽

Evolutionary Trend ◽

Ecological Evaluation ◽

Eutrophic Water ◽

Rrna Gene Sequencing ◽

Opposite Tendency

The effects of hydrodynamic disturbances on the bacterial communities in eutrophic aquatic environments remain poorly understood, despite their importance to ecological evaluation and remediation. This study investigated the evolution of bacterial communities in the water–sediment systems under the influence of three typical velocity conditions with the timescale of 5 weeks. The results demonstrated that higher bacterial diversity and notable differences were detected in sediment compared to water using the 16S rRNA gene sequencing. The phyla Firmicutes and γ-Proteobacteria survived better in both water and sediment under stronger water disturbances. Their relative abundance peaked at 36.0%, 33.2% in water and 38.0%, 43.6% in sediment, respectively, while the phylum Actinobacteria in water had the opposite tendency. Its relative abundance grew rapidly in static control (SC) and peaked at 44.8%, and it almost disappeared in disturbance conditions. These phenomena were caused by the proliferation of genus Exiguobacterium (belonging to Firmicutes), Citrobacter, Acinetobacter, Pseudomonas (belonging to γ-Proteobacteria), and hgcI_clade (belonging to Actinobacteria). The nonmetric multidimensional scaling (NMDS) and Venn analysis also revealed significantly different evolutionary trend in the three water-sediment systems. It was most likely caused by the changes of geochemical characteristics (dissolved oxygen (DO) and nutrients). This kind of study can provide helpful information for ecological assessment and remediation strategy in eutrophic aquatic environments.

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The female urinary microbiota in relation to the reproductive tract microbiota

10.1101/628974 ◽

2019 ◽

Author(s):

Chen Chen ◽

Lilan Hao ◽

Weixia Wei ◽

Fei Li ◽

Liju Song ◽

...

Keyword(s):

Microbial Communities ◽

Bacterial Communities ◽

Reproductive Tract ◽

Chinese Women ◽

Uterine Cavity ◽

Detailed Comparison ◽

Reproductive Age ◽

Rrna Gene ◽

Urogenital System ◽

Urinary Microbiota

AbstractHuman urine is traditionally considered to be sterile, and whether the urine harbours distinct microbial communities has been a matter of debate. The potential link between female urine and reproductive tract microbial communities is currently not clear.Here we collected the urine samples from 147 Chinese women of reproductive age, and explored the nature of colonization by 16S rRNA gene amplicon sequencing, real-time qPCR and live bacteria culture. To demonstrate utility intra-individual Spearman’s correlation was used to explore the relationship between urine and multi-sites of the reproductive tract. PERMANOVA was also performed to explore potential correlations between the lifestyle and various clinical factors and urinary bacterial communities. Our data demonstrated distinct bacterial communities in urine, indicative of a non-sterile environment. Types of diverse, Streptococcus-dominated, and Lactobacillus-dominated were the three most common types in the cohort. Detailed comparison of the urinary microbiota to the multi-sites of reproductive tract microbiota demonstrated the urinary microbiota was more similar to the microbiota in the cervix and uterine cavity instead of vagina in the same women.Our data demonstrates the potential connectivity of the microbiota in the female urogenital system and provided insight into the exploration of urethra and genital tract diseases.

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The leaf bacterial microbiota of female and male kiwifruit plants in distinct seasons: assessing the impact of Pseudomonas syringae pv. actinidiae

Phytobiomes Journal ◽

10.1094/pbiomes-09-20-0070-r ◽

2021 ◽

Author(s):

Aitana Ares ◽

Joana Pereira ◽

Eva Garcia ◽

Joana Costa ◽

Igor Tiago

Keyword(s):

Bacterial Community ◽

Pseudomonas Syringae ◽

Relative Abundance ◽

Bacterial Communities ◽

Defense Mechanisms ◽

Abiotic Factors ◽

Illumina Miseq ◽

Rrna Gene ◽

Sequencing Platform ◽

The Impact

The pandemic Pseudomonas syringae pv. actinidiae (Psa) has been compromising the production of the kiwifruit industry in major producing countries. Abiotic factors and plant gender are known to influence the disease outcome. To better understand their impact, we have determined the diversity of the leafs bacterial communities using the V5-V6 region of the 16S rRNA gene amplicon on the Illumina MiSeq sequencing platform. Healthy and diseased female and male kiwifruit plants were analyzed in two consecutive seasons: spring and autumn. This work describes whether the season, plant gender and the presence of Psa can affect the leaves bacterial community. Fifty bacterial operational taxonomic units (OTUs) were identified and assigned to five phyla distributed by 14 different families and 23 genera. The leaves of healthy female and male kiwi plants share most of the identified bacterial populations, that undergoes major seasonal changes. In both cases a substantial increase of the relative abundance of genus Methylobacterium is observed in autumn. The presence of Psa induced profound changes on leaves bacterial communities structure translated into a reduction in the relative abundance of previously dominant genera that had been found in healthy plants, namely Hymenobacter, Sphingomonas and Massilia. The impact of Psa was less pronounced in the bacterial community structure of male plants in both seasons. Some of the naturally occurring genera have the potential to act as an antagonist or as enhancers of the defense mechanisms paving the way for environmentally friendly and sustainable disease control.

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