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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 Micr...
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  
Rumen microbiology
키워드  
Fungi
키워드  
Genome-resolved metagenome
기타저자  
The Ohio State University Animal Sciences
기본자료저록  
Dissertations Abstracts International. 86-12B.
전자적 위치 및 접속  
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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.

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