High-Throughput Screening of Microbial Isolates with Impact on <em>Caenorhabditis elegans</em> Health

Irtiqa Ali, Jack Martin, Alejandra Zárate-Potes, Alexandre Benedetto

Research output: Contribution to journalArticle (journal)peer-review

2 Citations (Scopus)
1 Downloads (Pure)

Abstract

With its small size, short lifespan, and easy genetics, Caenorhabditis elegans offers a convenient platform to study the impact of microbial isolates on host physiology. It also fluoresces in blue when dying, providing a convenient means of pinpointing death. This property has been exploited to develop high-throughput label-free C. elegans survival assays (LFASS). These involve time-lapse fluorescence recording of worm populations set in multiwell plates, from which population median time of death can be derived. The present study adopts the LFASS approach to screen multiple microbial isolates at once for the effects on C. elegans susceptibility to severe heat and oxidative stresses. Such microbial screening pipeline, which can notably be used to prescreen probiotics, using severe stress resistance as a proxy for host health is reported here. The protocol describes how to grow both C. elegans gut microbiota isolate collections and synchronous worm populations in multiwell arrays before combining them for the assays. The example provided covers the testing of 47 bacterial isolates and one control strain on two worm strains, in two stress assays in parallel. However, the approach pipeline is readily scalable and applicable to the screening of many other modalities. Thus, it provides a versatile setup to rapidly survey a multiparametric landscape of biological and biochemical conditions that impact C. elegans health.
Original languageEnglish
Article numbere63860
JournalJournal of Visualized Experiments
Volume2022
Issue number182
Early online date28 Apr 2022
DOIs
Publication statusPublished - 28 Apr 2022

Keywords

  • High-Throughput Screening
  • Microbial Isolates
  • Caenorhabditis Elegans Health
  • Oxidative Thermal Stress Resistance
  • C.elegans Strains
  • Bacterial Isolates
  • Versatile And Scalable Approach
  • Rapid Screening
  • Conditions With Impact On C.elegans Health
  • Study Of Mechanisms Of Health And Diseases
  • Preclinical Screening Of Drugs
  • Antiemetics
  • Probiotics
  • Nematode Mutant
  • Knockdown
  • Transgenic
  • Disease Models
  • Complex Interactions
  • Host Microbe Interactions
  • Metabolic Disorders
  • Stress Handling
  • Aging
  • Contamination Prevention
  • E.coli OP50 Bacteria Culture Preparation

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