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Wastewater-Based and Environmental Surveillance of Infectious Diseases and Antimicrobial Resistance.
Wastewater-Based and Environmental Surveillance of Infectious Diseases and Antimicrobial R...
Wastewater-Based and Environmental Surveillance of Infectious Diseases and Antimicrobial Resistance.

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자료유형  
 학위논문(국외)
기본표목-개인명  
표제와 책임표시사항  
Wastewater-Based and Environmental Surveillance of Infectious Diseases and Antimicrobial Resistance.
발행, 배포, 간사 사항  
[S.l.] : University of Michigan. , 2025
    발행, 배포, 간사 사항  
    Ann Arbor : ProQuest Dissertations & Theses , 2025
      형태사항  
      164 p.
      일반주기  
      Source: Dissertations Abstracts International, Volume: 87-03, Section: B.
      일반주기  
      Advisor: Dvonch, Joseph Tim;Xi, Chuanwu.
      학위논문주기  
      Thesis (Ph.D.)--University of Michigan, 2025.
      요약 등 주기  
      요약Wastewater-based epidemiology (WBE) offers a powerful, non-invasive approach to monitor public health at the community level, especially in detecting infectious diseases and antimicrobial resistance (AMR). While WBE gained prominence during the COVID-19 pandemic, its broader application across environmental contexts and microbial threats remains underutilized. This dissertation addresses key methodological and knowledge gaps in WBE from multiple dimensions: Aim 1 develops high-throughput molecular tools for virus detection, Aim 2 implements AMR and Acinetobacter surveillance in urban wastewater by integrating multiple monitoring methods, and Aim 3 extends WBE principles and methods to environmental monitoring of surface water.Aim 1 focused on developing two multiplex reverse transcription droplet digital PCR (RT-ddPCR) panels to detect respiratory and enteric viruses (Influenza A/B, SARS-CoV-2, Norovirus GI/GII, RSV) in wastewater. The selected viruses represent major pathogens of public health concern that are frequently associated with community outbreaks and have been widely studied in wastewater-based surveillance. These multiplex RT-ddPCR panels were designed using the six-color QX600™ ddPCR system and validated using synthetic RNA and spiked wastewater samples. The multiplex assays demonstrated high specificity, sensitivity (LoD 0.25-0.75 copies/µL), and strong concordance with singleplex assays (R² 0.94, Kendall's τ 0.89), with minimal quantification bias. The results confirmed that multiplex RT-ddPCR offers a robust, efficient, and scalable method for viral surveillance, reducing sample and reagent usage while maintaining high analytical performance.Aim 2 investigated the spatial and temporal dynamics of AMR genes, mobile genetic elements (MGEs), and Acinetobacter spp. across a university wastewater network. Weekly samples were collected over 1.5 years from five locations, including a hospital outlet, residential sewers, and a treatment plant. To profile the microbiome and resistome of wastewater samples, selective culture of Acinetobacter spp., ddPCR for bacteria and AMR genes, and metagenomic sequencing were performed. We identified clear seasonal trends in bacterial abundance and location-specific differences in resistome and mobilome profiles. Acinetobacter spp. abundance correlated strongly with AMR gene load (r 0.7), suggesting that this genus plays a key role in shaping the resistome of urban wastewater. Genome-resolved metagenomics further revealed that dominant Acinetobacter genomes harbored clinically relevant resistance genes, such as blaOXA variants (conferring carbapenem resistance) and adeB/adeIJK (components of efflux pumps associated with multidrug resistance). This indicates that Acinetobacter not only carries a high burden of AMR genes but may also act as a primary reservoir and vehicle for their environmental persistence and potential spread.. Network analysis based on Spearman correlations between bacterial genera and AMR phenotypes identified Acinetobacter as a central hub, showing high degree, strength, and betweenness centrality compared to other genera. This indicates that Acinetobacter was one of the most strongly and broadly connected taxa in terms of co-occurrence with multiple resistance phenotypes, suggesting its prominent role in structuring the resistome within the wastewater system. Source tracking analysis suggested residential-like inputs, not hospital sources, as dominant contributors to the treatment plant influent. These findings demonstrate that wastewater surveillance can reveal hidden drivers of AMR dissemination-such as high-burden community sources-and identify key microbial taxa that contribute to resistance gene mobility. By integrating culture, molecular, and genomic approaches, this work provides a framework for monitoring AMR beyond clinical settings, informing targeted interventions in sewer-shed management, and supporting One Health efforts to mitigate resistance risks at the human-environment interface.Aim 3 applied WBE tools to assess fecal contamination in a recreational inland lake, extending WBE principles beyond centralized systems. Weekly samples during swimming seasons were tested for E. coli using culture and ddPCR (EC23S, AllBac), alongside MST markers for human (HF183) and goose (ND2) fecal sources. Results showed strong correlations between ddPCR and culture methods, with ddPCR offering greater sensitivity and reduced time to results. To identify contamination sources, we used multiple linear regression models, with fecal indicator bacteria (e.g., log-transformed E. coli, EC23S, or AllBac concentrations) as outcomes and MST marker concentrations as predictors. These models consistently identified ND2 (goose-associated) as the dominant predictor of fecal contamination, while HF183 (human-associated) contributions were minimal. While microbiologists have long monitored microbes in water systems, WBE methodologies represent a more integrated, population-level approach that combines quantitative molecular assays (like ddPCR), source attribution through MST markers, and frequent sampling to assess human health risks and contamination patterns in real time. In this study, WBE principles were adapted to a non-sewered setting by applying these molecular and statistical tools to track fecal pollution sources across space and time. These findings support targeted risk mitigation efforts-such as bird population management at recreational beaches-and demonstrate the broader adaptability of WBE frameworks for environmental surveillance in natural waters.Overall, this dissertation advances WBE by establishing and validating scalable, multi-target molecular tools and demonstrating their application across clinical, community, and recreational environments. By integrating ddPCR, metagenomics, and microbial source tracking, this research provides a holistic framework for monitoring pathogens, resistance genes, and fecal contamination. The findings have broad implications for environmental surveillance, public health protection, and One Health policy development, emphasizing the value of wastewater and environmental microbiology in addressing complex health threats at the human-environment interface.
      주제명부출표목-일반주제명  
      주제명부출표목-일반주제명  
      주제명부출표목-일반주제명  
      비통제 색인어  
      비통제 색인어  
      비통제 색인어  
      비통제 색인어  
      부출표목-단체명  
      University of Michigan Environmental Health Sciences
        기본자료저록  
        Dissertations Abstracts International. 87-03B.
        전자적 위치 및 접속  
         원문정보보기

        MARC

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        ■040    ▼aMiAaPQ▼cMiAaPQ
        ■0820  ▼a614
        ■1001  ▼aLi,  Xin.
        ■24510▼aWastewater-Based  and  Environmental  Surveillance  of  Infectious  Diseases  and  Antimicrobial  Resistance.
        ■260    ▼a[S.l.]▼bUniversity  of  Michigan.  ▼c2025
        ■260  1▼aAnn  Arbor▼bProQuest  Dissertations  &  Theses▼c2025
        ■300    ▼a164  p.
        ■500    ▼aSource:  Dissertations  Abstracts  International,  Volume:  87-03,  Section:  B.
        ■500    ▼aAdvisor:  Dvonch,  Joseph  Tim;Xi,  Chuanwu.
        ■5021  ▼aThesis  (Ph.D.)--University  of  Michigan,  2025.
        ■520    ▼aWastewater-based  epidemiology  (WBE)  offers  a  powerful,  non-invasive  approach  to  monitor  public  health  at  the  community  level,  especially  in  detecting  infectious  diseases  and  antimicrobial  resistance  (AMR).  While  WBE  gained  prominence  during  the  COVID-19  pandemic,  its  broader  application  across  environmental  contexts  and  microbial  threats  remains  underutilized.  This  dissertation  addresses  key  methodological  and  knowledge  gaps  in  WBE  from  multiple  dimensions:  Aim  1  develops  high-throughput  molecular  tools  for  virus  detection,  Aim  2  implements  AMR  and  Acinetobacter  surveillance  in  urban  wastewater  by  integrating  multiple  monitoring  methods,  and  Aim  3  extends  WBE  principles  and  methods  to  environmental  monitoring  of  surface  water.Aim  1  focused  on  developing  two  multiplex  reverse  transcription  droplet  digital  PCR  (RT-ddPCR)  panels  to  detect  respiratory  and  enteric  viruses  (Influenza  A/B,  SARS-CoV-2,  Norovirus  GI/GII,  RSV)  in  wastewater.  The  selected  viruses  represent  major  pathogens  of  public  health  concern  that  are  frequently  associated  with  community  outbreaks  and  have  been  widely  studied  in  wastewater-based  surveillance.  These  multiplex  RT-ddPCR  panels  were  designed  using  the  six-color  QX600™  ddPCR  system  and  validated  using  synthetic  RNA  and  spiked  wastewater  samples.  The  multiplex  assays  demonstrated  high  specificity,  sensitivity  (LoD  0.25-0.75  copies/µL),  and  strong  concordance  with  singleplex  assays  (R²    0.94,  Kendall's  τ    0.89),  with  minimal  quantification  bias.  The  results  confirmed  that  multiplex  RT-ddPCR  offers  a  robust,  efficient,  and  scalable  method  for  viral  surveillance,  reducing  sample  and  reagent  usage  while  maintaining  high  analytical  performance.Aim  2  investigated  the  spatial  and  temporal  dynamics  of  AMR  genes,  mobile  genetic  elements  (MGEs),  and  Acinetobacter  spp.  across  a  university  wastewater  network.  Weekly  samples  were  collected  over  1.5  years  from  five  locations,  including  a  hospital  outlet,  residential  sewers,  and  a  treatment  plant.  To  profile  the  microbiome  and  resistome  of  wastewater  samples,  selective  culture  of  Acinetobacter  spp.,  ddPCR  for  bacteria  and  AMR  genes,  and  metagenomic  sequencing  were  performed.  We  identified  clear  seasonal  trends  in  bacterial  abundance  and  location-specific  differences  in  resistome  and  mobilome  profiles.  Acinetobacter  spp.  abundance  correlated  strongly  with  AMR  gene  load  (r    0.7),  suggesting  that  this  genus  plays  a  key  role  in  shaping  the  resistome  of  urban  wastewater.  Genome-resolved  metagenomics  further  revealed  that  dominant  Acinetobacter  genomes  harbored  clinically  relevant  resistance  genes,  such  as  blaOXA  variants  (conferring  carbapenem  resistance)  and  adeB/adeIJK  (components  of  efflux  pumps  associated  with  multidrug  resistance).  This  indicates  that  Acinetobacter  not  only  carries  a  high  burden  of  AMR  genes  but  may  also  act  as  a  primary  reservoir  and  vehicle  for  their  environmental  persistence  and  potential  spread..  Network  analysis  based  on  Spearman  correlations  between  bacterial  genera  and  AMR  phenotypes  identified  Acinetobacter  as  a  central  hub,  showing  high  degree,  strength,  and  betweenness  centrality  compared  to  other  genera.  This  indicates  that  Acinetobacter  was  one  of  the  most  strongly  and  broadly  connected  taxa  in  terms  of  co-occurrence  with  multiple  resistance  phenotypes,  suggesting  its  prominent  role  in  structuring  the  resistome  within  the  wastewater  system.  Source  tracking  analysis  suggested  residential-like  inputs,  not  hospital  sources,  as  dominant  contributors  to  the  treatment  plant  influent.  These  findings  demonstrate  that  wastewater  surveillance  can  reveal  hidden  drivers  of  AMR  dissemination-such  as  high-burden  community  sources-and  identify  key  microbial  taxa  that  contribute  to  resistance  gene  mobility.  By  integrating  culture,  molecular,  and  genomic  approaches,  this  work  provides  a  framework  for  monitoring  AMR  beyond  clinical  settings,  informing  targeted  interventions  in  sewer-shed  management,  and  supporting  One  Health  efforts  to  mitigate  resistance  risks  at  the  human-environment  interface.Aim  3  applied  WBE  tools  to  assess  fecal  contamination  in  a  recreational  inland  lake,  extending  WBE  principles  beyond  centralized  systems.  Weekly  samples  during  swimming  seasons  were  tested  for  E.  coli  using  culture  and  ddPCR  (EC23S,  AllBac),  alongside  MST  markers  for  human  (HF183)  and  goose  (ND2)  fecal  sources.  Results  showed  strong  correlations  between  ddPCR  and  culture  methods,  with  ddPCR  offering  greater  sensitivity  and  reduced  time  to  results.  To  identify  contamination  sources,  we  used  multiple  linear  regression  models,  with  fecal  indicator  bacteria  (e.g.,  log-transformed  E.  coli,  EC23S,  or  AllBac  concentrations)  as  outcomes  and  MST  marker  concentrations  as  predictors.  These  models  consistently  identified  ND2  (goose-associated)  as  the  dominant  predictor  of  fecal  contamination,  while  HF183  (human-associated)  contributions  were  minimal.  While  microbiologists  have  long  monitored  microbes  in  water  systems,  WBE  methodologies  represent  a  more  integrated,  population-level  approach  that combines  quantitative  molecular  assays  (like  ddPCR),  source  attribution  through  MST  markers,  and  frequent  sampling  to  assess  human  health  risks  and  contamination  patterns  in  real  time.  In  this  study,  WBE  principles  were  adapted  to  a  non-sewered  setting  by  applying  these  molecular  and  statistical  tools  to  track  fecal  pollution  sources  across  space  and  time.  These  findings  support  targeted  risk  mitigation  efforts-such  as  bird  population  management  at  recreational  beaches-and  demonstrate  the  broader  adaptability  of  WBE  frameworks  for  environmental  surveillance  in  natural  waters.Overall,  this  dissertation  advances  WBE  by  establishing  and  validating  scalable,  multi-target  molecular  tools  and  demonstrating  their  application  across  clinical,  community,  and  recreational  environments.  By  integrating  ddPCR,  metagenomics,  and  microbial  source  tracking,  this  research  provides  a  holistic  framework  for  monitoring  pathogens,  resistance  genes,  and  fecal  contamination.  The  findings  have  broad  implications  for  environmental  surveillance,  public  health  protection,  and  One  Health  policy  development,  emphasizing  the  value  of  wastewater  and  environmental  microbiology  in  addressing  complex  health  threats  at  the  human-environment  interface.
        ■590    ▼aSchool  code:  0127.
        ■650  4▼aPublic  health.
        ■650  4▼aEnvironmental  health.
        ■650  4▼aEnvironmental  science.
        ■653    ▼aWastewater-based  epidemiology
        ■653    ▼aAntimicrobial  resistance
        ■653    ▼aMicrobial  source  tracking
        ■653    ▼aCOVID-19
        ■690    ▼a0470
        ■690    ▼a0573
        ■690    ▼a0768
        ■71020▼aUniversity  of  Michigan▼bEnvironmental  Health  Sciences.
        ■7730  ▼tDissertations  Abstracts  International▼g87-03B.
        ■790    ▼a0127
        ■791    ▼aPh.D.
        ■792    ▼a2025
        ■793    ▼aEnglish
        ■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17359872▼nKERIS▼z이  자료의  원문은  한국교육학술정보원에서  제공합니다.

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