Sequencing and culture-based characterization of the vaginal and uterine microbiota in beef cattle that became pregnant or remained open following artificial insemination

ABSTRACT In this study, we evaluated the vaginal and uterine microbiota between beef cattle that became pregnant via artificial insemination (AI) and those that remained open to identify microbial signatures associated with fertility. We also characterized the culturable fraction of these microbiota...

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Main Authors: Emily M. Webb, Devin B. Holman, Kaycie N. Schmidt, Beena Pun, Kevin K. Sedivec, Jennifer L. Hurlbert, Kerri A. Bochantin, Alison K. Ward, Carl R. Dahlen, Samat Amat
Format: Article
Language:English
Published: American Society for Microbiology 2023-12-01
Series:Microbiology Spectrum
Subjects:
Online Access:https://journals.asm.org/doi/10.1128/spectrum.02732-23
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author Emily M. Webb
Devin B. Holman
Kaycie N. Schmidt
Beena Pun
Kevin K. Sedivec
Jennifer L. Hurlbert
Kerri A. Bochantin
Alison K. Ward
Carl R. Dahlen
Samat Amat
author_facet Emily M. Webb
Devin B. Holman
Kaycie N. Schmidt
Beena Pun
Kevin K. Sedivec
Jennifer L. Hurlbert
Kerri A. Bochantin
Alison K. Ward
Carl R. Dahlen
Samat Amat
author_sort Emily M. Webb
collection DOAJ
description ABSTRACT In this study, we evaluated the vaginal and uterine microbiota between beef cattle that became pregnant via artificial insemination (AI) and those that remained open to identify microbial signatures associated with fertility. We also characterized the culturable fraction of these microbiota and screened certain vaginal and uterine bacterial isolates for antimicrobial resistance. For this, vaginal and uterine swabs were collected before AI from two cohorts of Angus-crossbred cattle: mature cows (vaginal and uterine: 27 open and 31 pregnant) and heifers (vaginal: 26 open and 33 pregnant). The microbiota of these samples were assessed using 16S rRNA gene sequencing and culturing. Twenty-nine vaginal and uterine bacterial isolates were screened for resistance against 41 antibiotics. Within the vaginal microbiota, 11 amplicon sequence variants (ASVs) were more relatively abundant in the open heifers compared with the pregnant heifers. No differentially abundant ASVs were detected in the vaginal samples from pregnant versus open cows. Pregnant cows had a distinct uterine microbiota community structure (P = 0.008) and interaction network structure compared with open cows. Twenty-eight differentially abundant uterine ASVs were observed between the pregnant and open cows. Community structure and diversity were different between the cow’s vagina and uterus. A total of 733 bacterial isolates were recovered from vaginal (512) and uterine (221) swabs under aerobic (83 different species) and anaerobic (69 species) culturing. Among these isolates were pathogenic species and those mostly susceptible to tested antibiotics. Overall, our results indicate that fertility-associated taxonomic signatures are present in the bovine uterine and vaginal microbiota. IMPORTANCE Emerging evidence suggests that microbiome-targeted approaches may provide a novel opportunity to reduce the incidence of reproductive failures in cattle. To develop such microbiome-based strategies, one of the first logical steps is to identify reproductive microbiome features related to fertility and to isolate the fertility-associated microbial species for developing a future bacterial consortium that could be administered before breeding to enhance pregnancy outcomes. Here, we characterized the vaginal and uterine microbiota in beef cattle that became pregnant or remained open via artificial insemination and identified microbiota features associated with fertility. We compared similarities between vaginal and uterine microbiota and between heifers and cows. Using culturing, we provided new insights into the culturable fraction of the vaginal and uterine microbiota and their antimicrobial resistance. Overall, our findings will serve as an important basis for future research aimed at harnessing the vaginal and uterine microbiome for improved cattle fertility.
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spelling doaj.art-403e5987c5ae474d99f2ba46c92c897f2023-12-12T13:17:18ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972023-12-0111610.1128/spectrum.02732-23Sequencing and culture-based characterization of the vaginal and uterine microbiota in beef cattle that became pregnant or remained open following artificial inseminationEmily M. Webb0Devin B. Holman1Kaycie N. Schmidt2Beena Pun3Kevin K. Sedivec4Jennifer L. Hurlbert5Kerri A. Bochantin6Alison K. Ward7Carl R. Dahlen8Samat Amat9Department of Microbiological Sciences, North Dakota State University , Fargo, North Dakota, USALacombe Research and Development Centre, Agriculture and Agri-Food Canada , Lacombe, Alberta, CanadaDepartment of Microbiological Sciences, North Dakota State University , Fargo, North Dakota, USADepartment of Microbiological Sciences, North Dakota State University , Fargo, North Dakota, USACentral Grasslands Research Extension Center, North Dakota State University , Streeter, North Dakota, USADepartment of Animal Sciences and Center for Nutrition and Pregnancy, North Dakota State University , Fargo, North Dakota, USADepartment of Animal Sciences and Center for Nutrition and Pregnancy, North Dakota State University , Fargo, North Dakota, USADepartment of Animal Sciences and Center for Nutrition and Pregnancy, North Dakota State University , Fargo, North Dakota, USADepartment of Animal Sciences and Center for Nutrition and Pregnancy, North Dakota State University , Fargo, North Dakota, USADepartment of Microbiological Sciences, North Dakota State University , Fargo, North Dakota, USAABSTRACT In this study, we evaluated the vaginal and uterine microbiota between beef cattle that became pregnant via artificial insemination (AI) and those that remained open to identify microbial signatures associated with fertility. We also characterized the culturable fraction of these microbiota and screened certain vaginal and uterine bacterial isolates for antimicrobial resistance. For this, vaginal and uterine swabs were collected before AI from two cohorts of Angus-crossbred cattle: mature cows (vaginal and uterine: 27 open and 31 pregnant) and heifers (vaginal: 26 open and 33 pregnant). The microbiota of these samples were assessed using 16S rRNA gene sequencing and culturing. Twenty-nine vaginal and uterine bacterial isolates were screened for resistance against 41 antibiotics. Within the vaginal microbiota, 11 amplicon sequence variants (ASVs) were more relatively abundant in the open heifers compared with the pregnant heifers. No differentially abundant ASVs were detected in the vaginal samples from pregnant versus open cows. Pregnant cows had a distinct uterine microbiota community structure (P = 0.008) and interaction network structure compared with open cows. Twenty-eight differentially abundant uterine ASVs were observed between the pregnant and open cows. Community structure and diversity were different between the cow’s vagina and uterus. A total of 733 bacterial isolates were recovered from vaginal (512) and uterine (221) swabs under aerobic (83 different species) and anaerobic (69 species) culturing. Among these isolates were pathogenic species and those mostly susceptible to tested antibiotics. Overall, our results indicate that fertility-associated taxonomic signatures are present in the bovine uterine and vaginal microbiota. IMPORTANCE Emerging evidence suggests that microbiome-targeted approaches may provide a novel opportunity to reduce the incidence of reproductive failures in cattle. To develop such microbiome-based strategies, one of the first logical steps is to identify reproductive microbiome features related to fertility and to isolate the fertility-associated microbial species for developing a future bacterial consortium that could be administered before breeding to enhance pregnancy outcomes. Here, we characterized the vaginal and uterine microbiota in beef cattle that became pregnant or remained open via artificial insemination and identified microbiota features associated with fertility. We compared similarities between vaginal and uterine microbiota and between heifers and cows. Using culturing, we provided new insights into the culturable fraction of the vaginal and uterine microbiota and their antimicrobial resistance. Overall, our findings will serve as an important basis for future research aimed at harnessing the vaginal and uterine microbiome for improved cattle fertility.https://journals.asm.org/doi/10.1128/spectrum.02732-23vaginal microbiotauterine microbiotabovine16S rRNA gene sequencingculturingpregnancy
spellingShingle Emily M. Webb
Devin B. Holman
Kaycie N. Schmidt
Beena Pun
Kevin K. Sedivec
Jennifer L. Hurlbert
Kerri A. Bochantin
Alison K. Ward
Carl R. Dahlen
Samat Amat
Sequencing and culture-based characterization of the vaginal and uterine microbiota in beef cattle that became pregnant or remained open following artificial insemination
Microbiology Spectrum
vaginal microbiota
uterine microbiota
bovine
16S rRNA gene sequencing
culturing
pregnancy
title Sequencing and culture-based characterization of the vaginal and uterine microbiota in beef cattle that became pregnant or remained open following artificial insemination
title_full Sequencing and culture-based characterization of the vaginal and uterine microbiota in beef cattle that became pregnant or remained open following artificial insemination
title_fullStr Sequencing and culture-based characterization of the vaginal and uterine microbiota in beef cattle that became pregnant or remained open following artificial insemination
title_full_unstemmed Sequencing and culture-based characterization of the vaginal and uterine microbiota in beef cattle that became pregnant or remained open following artificial insemination
title_short Sequencing and culture-based characterization of the vaginal and uterine microbiota in beef cattle that became pregnant or remained open following artificial insemination
title_sort sequencing and culture based characterization of the vaginal and uterine microbiota in beef cattle that became pregnant or remained open following artificial insemination
topic vaginal microbiota
uterine microbiota
bovine
16S rRNA gene sequencing
culturing
pregnancy
url https://journals.asm.org/doi/10.1128/spectrum.02732-23
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