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Quantification of the capacity of vibrio fischeri to establish symbiosis with Euprymna scolopes.
Donnelly, Aidan R; Giacobe, Elizabeth J; Cook, Rachel A; Francis, Gareth M; Buddle, Grace K; Beaubrun, Christina L; Cecere, Andrew G; Miyashiro, Tim I.
Afiliação
  • Donnelly AR; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, United States of America.
  • Giacobe EJ; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, United States of America.
  • Cook RA; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, United States of America.
  • Francis GM; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, United States of America.
  • Buddle GK; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, United States of America.
  • Beaubrun CL; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, United States of America.
  • Cecere AG; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, United States of America.
  • Miyashiro TI; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, United States of America.
PLoS One ; 18(7): e0287519, 2023.
Article em En | MEDLINE | ID: mdl-37440554
Most animals establish long-term symbiotic associations with bacteria that are critical for normal host physiology. The symbiosis that forms between the Hawaiian squid Euprymna scolopes and the bioluminescent bacterium Vibrio fischeri serves as an important model system for investigating the molecular mechanisms that promote animal-bacterial symbioses. E. scolopes hatch from their eggs uncolonized, which has led to the development of squid-colonization assays that are based on introducing culture-grown V. fischeri cells to freshly hatched juvenile squid. Recent studies have revealed that strains often exhibit large differences in how they establish symbiosis. Therefore, we sought to develop a simplified and reproducible protocol that permits researchers to determine appropriate inoculum levels and provides a platform to standardize the assay across different laboratories. In our protocol, we adapt a method commonly used for evaluating the infectivity of pathogens to quantify the symbiotic capacity of V. fischeri strains. The resulting metric, the symbiotic dose-50 (SD50), estimates the inoculum level that is necessary for a specific V. fischeri strain to establish a light-emitting symbiosis. Relative to other protocols, our method requires 2-5-fold fewer animals. Furthermore, the power analysis presented here suggests that the protocol can detect up to a 3-fold change in the SD50 between different strains.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Vibrio / Aliivibrio fischeri Idioma: En Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Vibrio / Aliivibrio fischeri Idioma: En Ano de publicação: 2023 Tipo de documento: Article