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Elucidating the Under-Explored Genomic Diversity and Metabolic Potential of the Rumen Microbiome Through Multi-Omics Approaches
Elucidating the Under-Explored Genomic Diversity and Metabolic Potential of the Rumen Microbiome Through Multi-Omics Approaches
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20260202104757
- ISBN
- 9798280770171
- DDC
- 574
- 저자명
- Yan, Ming.
- 서명/저자
- Elucidating the Under-Explored Genomic Diversity and Metabolic Potential of the Rumen Microbiome Through Multi-Omics Approaches
- 발행사항
- [Sl] : The Ohio State University, 2024
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- 형태사항
- 237 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 86-12, Section: B.
- 주기사항
- Advisor: Yu, Zhongtang.
- 학위논문주기
- Thesis (Ph.D.)--The Ohio State University, 2024.
- 초록/해제
- 요약The rumen hosts a diverse array of prokaryotic (bacteria and archaea) and eukaryotic (fungi and protozoa) microbes. Collectively, they hydrolyze complex plant cell wall materials into simple sugars, which are further fermented into VFA, representing a substantial source of the host's energy needs. By incorporating inorganic ammonia generated from feed protein and urea, rumen microbes also provide a significant portion of the host's protein requirements. As regulators of the microbial ecosystem, rumen viruses (bacteriophages and eukaryotic viruses) also influence rumen fermentation and microbial protein synthesis. They achieve this by directly lysing microbes, thereby modulating microbial composition or by modifying the metabolism of infected bacterial cells. Additionally, they drive co-evolution between microbes and viruses, acting as vectors for horizontal gene transfer or through dynamic defense and counterdefense interactions with microbes.The anaerobic microbial cultivation techniques developed by Robert Hungate enable rumen microbiologists to explore the diverse spectrum of rumen microbial physiology and metabolism. However, despite continuous efforts in anaerobic cultivation, the culturable rumen microbes (including viruses) represent only a limited fraction of the overall diversity. Moreover, microbial cultures, whether monocultures or mixed cultures, fail to fully replicate the intricate microbial interactions observed in vivo, such as cross-feeding and predatory conditions. Fortunately, multi-omics technologies complement traditional culture-dependent analyses, enabling us to explore microbial ecology by uncovering the genomes (via genome-resolved metagenomics) and metabolism (via metatranscriptomics, metaproteomics and enzymatic activities) of the unculturable majority. Utilizing advancements in multi-omics and bioinformatics, this research aims to bridge the gap in rumen microbial genomics within the context of microbial ecology and rumen fermentation.The first objective of this research (Chapter 3) is to explore the largely underexplored rumen phage genomics, providing the basic framework of rumen phage diversity, taxonomy, and functional potential. By analyzing 975 rumen metagenomes collected from 13 ruminant species (both domestic and wild) across 5 continents, we have identified a vast diversity of phages in the rumen that impact all major rumen microbes. This effort culminated in the establishment of the first comprehensive rumen virome database (RVD), significantly enhancing the identification of phage sequences in rumen metagenomic data compared to existing databases. Through detailed genomic analyses of these phage sequences, we discovered a variety of rumen phages encoding diverse auxiliary metabolic genes (AMGs) capable of directly modifying their host's central metabolism. Additionally, we found direct evidence of phages serving as reservoirs of antimicrobial resistance genes (ARGs). Together, the RVD database and functional framework established here provide a solid foundation for further studies on rumen viruses.The second objective of this research (Chapter 4) is to investigate the ecological significance of rumen phages, focusing on their role in microbial assembly and impact on microbial-phage interactions in relation to animal production. Our research initially focused on identifying prophage sequences within rumen microbial genomes and investigating their role in driving microbial evolution. Subsequently, we analyzed microbe-phage infection networks and co-occurrence networks to assess the significance of rumen phages in shaping microbial structures at both the strain and community levels. Finally, we conducted a reanalysis of data reported by nine studies involving treatment groups of ruminants with varying animal efficiency or dietary compositions to understand the impact of phage diversity and microbe-phage interactions on animal production. Collectively, we found that prophages are common in the rumen microbial genomes and may drive microbial diversification. In addition, viruses in the rumen may regulate microbes at strain and community levels through both antagonistic and mutualistic (through lysogeny) interactions. Moreover, this study establishes that the rumen virome demonstrates responsiveness to dietary shifts and associations with key animal production traits, including feed efficiency, lactation performance, weight gain, and methane emissions.The third objective of this research (Chapter 5) is to develop a bioinformatics program to identify sequences of eukaryotic microbes (protozoa and fungi) from rumen metagenomes and analyze the genomic and functional diversity of the newly identified sequences at both contig and bin levels. Using our newly developed program (referred to as GutEuk), we analyzed thousands of assembled genomes and tens of thousands of contigs to identify fungal and protozoan sequences. We conducted phylogenetic analysis with the newly identified genomes, along with previously established genomes, to explore potential novel diversity. Additionally, we predicted and annotated protein sequences from the newly identified rumen microbial eukaryotes and evaluated the program's effectiveness in enhancing metaproteomics analysis. Overall, GutEuk demonstrates improved performance in identifying microbial eukaryotes in the rumen and reveals an unexplored protozoan diversity. The established protein sequence database can improve protein identification from rumen metaproteomics, which is often a limiting factor in such studies.The fourth objective of this research (Chapter 6) is to investigate microbial N utilization in the rumen and their regulation under different energy and nitrogen availability. Firstly, we collected thousands of rumen microbial genomes and conduct comparative genomics analysis to identify the species-specific N assimilation and polysaccharide degradation paradigms. We also reanalyzed RVD to identify rumen phages that encode N assimilation-rated genes as AMGs. Then, we conducted an animal trial with 11 pairs of lamb twins using a split-plot (concentrate level as the main plot and crude protein level as the subplot) crossover design (different crude protein level over two periods) as a model to study the rumen microbial compositional change at different forage and CP levels. We identified potential nitrogen utilization strategies for hundreds of rumen microbes based on the genome annotation. In the absence of a complete glutamine synthetase-glutamate synthase pathway, many of the examined rumen microbes may rely on glutamate dehydrogenase for ammonia assimilation. Additionally, they may utilize other enzymes such as aspartase and alanine dehydrogenase, as well as preformed amino acids and peptides, to meet their nitrogen requirements. We also identified rumen microbes potentially involved in denitrification and urea hydrolysis processes. The classic nitrogen assimilation regulatory pathway was incomplete in most of the examined genomes, indicating the presence of alternative regulatory mechanisms. Ecological analysis supported the genome annotation findings, suggesting that ammonia concentration may not be a limiting factor in the rumen. Instead, rumen microbes appear to employ diverse, species-specific nitrogen utilization strategies in response to fluctuations in energy and nitrogen levels within the rumen. Overall, the rumen microbial nitrogen utilization strategies identified in this study, including those extending to unculturable microbes, could facilitate future practical research aimed at optimizing dietary formulations and modeling to enhance nitrogen utilization efficiency in ruminants.
- 일반주제명
- Bioinformatics
- 일반주제명
- Microbiology
- 일반주제명
- Animal sciences
- 일반주제명
- Genetics
- 키워드
- Rumen microbiome
- 키워드
- Fungi
- 기타저자
- The Ohio State University Animal Sciences
- 기본자료저록
- Dissertations Abstracts International. 86-12B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■1001 ▼aYan, Ming.
■24510▼aElucidating the Under-Explored Genomic Diversity and Metabolic Potential of the Rumen Microbiome Through Multi-Omics Approaches
■260 ▼a[Sl]▼bThe Ohio State University▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a237 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 86-12, Section: B.
■500 ▼aAdvisor: Yu, Zhongtang.
■5021 ▼aThesis (Ph.D.)--The Ohio State University, 2024.
■520 ▼aThe rumen hosts a diverse array of prokaryotic (bacteria and archaea) and eukaryotic (fungi and protozoa) microbes. Collectively, they hydrolyze complex plant cell wall materials into simple sugars, which are further fermented into VFA, representing a substantial source of the host's energy needs. By incorporating inorganic ammonia generated from feed protein and urea, rumen microbes also provide a significant portion of the host's protein requirements. As regulators of the microbial ecosystem, rumen viruses (bacteriophages and eukaryotic viruses) also influence rumen fermentation and microbial protein synthesis. They achieve this by directly lysing microbes, thereby modulating microbial composition or by modifying the metabolism of infected bacterial cells. Additionally, they drive co-evolution between microbes and viruses, acting as vectors for horizontal gene transfer or through dynamic defense and counterdefense interactions with microbes.The anaerobic microbial cultivation techniques developed by Robert Hungate enable rumen microbiologists to explore the diverse spectrum of rumen microbial physiology and metabolism. However, despite continuous efforts in anaerobic cultivation, the culturable rumen microbes (including viruses) represent only a limited fraction of the overall diversity. Moreover, microbial cultures, whether monocultures or mixed cultures, fail to fully replicate the intricate microbial interactions observed in vivo, such as cross-feeding and predatory conditions. Fortunately, multi-omics technologies complement traditional culture-dependent analyses, enabling us to explore microbial ecology by uncovering the genomes (via genome-resolved metagenomics) and metabolism (via metatranscriptomics, metaproteomics and enzymatic activities) of the unculturable majority. Utilizing advancements in multi-omics and bioinformatics, this research aims to bridge the gap in rumen microbial genomics within the context of microbial ecology and rumen fermentation.The first objective of this research (Chapter 3) is to explore the largely underexplored rumen phage genomics, providing the basic framework of rumen phage diversity, taxonomy, and functional potential. By analyzing 975 rumen metagenomes collected from 13 ruminant species (both domestic and wild) across 5 continents, we have identified a vast diversity of phages in the rumen that impact all major rumen microbes. This effort culminated in the establishment of the first comprehensive rumen virome database (RVD), significantly enhancing the identification of phage sequences in rumen metagenomic data compared to existing databases. Through detailed genomic analyses of these phage sequences, we discovered a variety of rumen phages encoding diverse auxiliary metabolic genes (AMGs) capable of directly modifying their host's central metabolism. Additionally, we found direct evidence of phages serving as reservoirs of antimicrobial resistance genes (ARGs). Together, the RVD database and functional framework established here provide a solid foundation for further studies on rumen viruses.The second objective of this research (Chapter 4) is to investigate the ecological significance of rumen phages, focusing on their role in microbial assembly and impact on microbial-phage interactions in relation to animal production. Our research initially focused on identifying prophage sequences within rumen microbial genomes and investigating their role in driving microbial evolution. Subsequently, we analyzed microbe-phage infection networks and co-occurrence networks to assess the significance of rumen phages in shaping microbial structures at both the strain and community levels. Finally, we conducted a reanalysis of data reported by nine studies involving treatment groups of ruminants with varying animal efficiency or dietary compositions to understand the impact of phage diversity and microbe-phage interactions on animal production. Collectively, we found that prophages are common in the rumen microbial genomes and may drive microbial diversification. In addition, viruses in the rumen may regulate microbes at strain and community levels through both antagonistic and mutualistic (through lysogeny) interactions. Moreover, this study establishes that the rumen virome demonstrates responsiveness to dietary shifts and associations with key animal production traits, including feed efficiency, lactation performance, weight gain, and methane emissions.The third objective of this research (Chapter 5) is to develop a bioinformatics program to identify sequences of eukaryotic microbes (protozoa and fungi) from rumen metagenomes and analyze the genomic and functional diversity of the newly identified sequences at both contig and bin levels. Using our newly developed program (referred to as GutEuk), we analyzed thousands of assembled genomes and tens of thousands of contigs to identify fungal and protozoan sequences. We conducted phylogenetic analysis with the newly identified genomes, along with previously established genomes, to explore potential novel diversity. Additionally, we predicted and annotated protein sequences from the newly identified rumen microbial eukaryotes and evaluated the program's effectiveness in enhancing metaproteomics analysis. Overall, GutEuk demonstrates improved performance in identifying microbial eukaryotes in the rumen and reveals an unexplored protozoan diversity. The established protein sequence database can improve protein identification from rumen metaproteomics, which is often a limiting factor in such studies.The fourth objective of this research (Chapter 6) is to investigate microbial N utilization in the rumen and their regulation under different energy and nitrogen availability. Firstly, we collected thousands of rumen microbial genomes and conduct comparative genomics analysis to identify the species-specific N assimilation and polysaccharide degradation paradigms. We also reanalyzed RVD to identify rumen phages that encode N assimilation-rated genes as AMGs. Then, we conducted an animal trial with 11 pairs of lamb twins using a split-plot (concentrate level as the main plot and crude protein level as the subplot) crossover design (different crude protein level over two periods) as a model to study the rumen microbial compositional change at different forage and CP levels. We identified potential nitrogen utilization strategies for hundreds of rumen microbes based on the genome annotation. In the absence of a complete glutamine synthetase-glutamate synthase pathway, many of the examined rumen microbes may rely on glutamate dehydrogenase for ammonia assimilation. Additionally, they may utilize other enzymes such as aspartase and alanine dehydrogenase, as well as preformed amino acids and peptides, to meet their nitrogen requirements. We also identified rumen microbes potentially involved in denitrification and urea hydrolysis processes. The classic nitrogen assimilation regulatory pathway was incomplete in most of the examined genomes, indicating the presence of alternative regulatory mechanisms. Ecological analysis supported the genome annotation findings, suggesting that ammonia concentration may not be a limiting factor in the rumen. Instead, rumen microbes appear to employ diverse, species-specific nitrogen utilization strategies in response to fluctuations in energy and nitrogen levels within the rumen. Overall, the rumen microbial nitrogen utilization strategies identified in this study, including those extending to unculturable microbes, could facilitate future practical research aimed at optimizing dietary formulations and modeling to enhance nitrogen utilization efficiency in ruminants.
■590 ▼aSchool code: 0168.
■650 4▼aBioinformatics
■650 4▼aMicrobiology
■650 4▼aAnimal sciences
■650 4▼aGenetics
■653 ▼aRumen microbiome
■653 ▼aRumen microbiology
■653 ▼aFungi
■653 ▼aGenome-resolved metagenome
■690 ▼a0475
■690 ▼a0715
■690 ▼a0410
■690 ▼a0369
■71020▼aThe Ohio State University▼bAnimal Sciences.
■7730 ▼tDissertations Abstracts International▼g86-12B.
■790 ▼a0168
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17358828▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.


