Jun 15, 2022

Public workspaceProtocol for Bulk Water Enrichment for the Isolation of Salmonella from Surface Water

DOI
dx.doi.org/10.17504/protocols.io.ewov1n3xygr2/v1
  • 1USDA ARS Environmental Microbial and Food Safety Laboratory;
  • 2USDA Agricultural Research Service Environmental Microbial and Food Safety Laboratory;
  • 3USDA Agricultural Research Service Meat Safety and Quality Laboratory;
  • 4USDA Agricultural Research Service Water Efficiency and Salinity Research Unit;
  • 5National Antimicrobial Resistance Monitoring System Environmental Working Group;
  • 6USDA Agricultural Research Service
  • NARMS EWG: Lisa Durso, Lari Hiott, Cheryl L East, Betty McConn, Andrea Ottesen, Jie Zheng, Laura Boczek;
Manan Sharma
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DOI: dx.doi.org/10.17504/protocols.io.ewov1n3xygr2/v1
Protocol Citation: Autumn L Kraft, Manan Sharma, James E Wells, Abasiofiok Mark Ibekwe, NARMS EWG, Jonathan G Frye 2022. Protocol for Bulk Water Enrichment for the Isolation of Salmonella from Surface Water. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov1n3xygr2/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
This protocol was used to examine 4 different sources of water inoculated with a low level of Salmonella Typhimurium (ca 0.030 CFU/mL).
Created: May 04, 2022
Last Modified: April 19, 2023
Protocol Integer ID: 61969
Keywords: National Program 108, Environmental Microbial and Food Safety Laboratory, Sureface water, Salmonella, water enrichment
Abstract
This protocol details the bulk water enrichment for the isolation of Salmonella from surface water.
Attachments
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411-892.docx
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Materials
Supplies Needed

  • Sterilized 2 L bottles
  • Sterilized 500 mL and 1L graduated cylinders
  • 5x Universal Pre-Enrichment broth (UPB, Acumedia #7510)
  • Control strain, Salmonella typhimurium BIOBALL® Luminate (BioMerieux, #422190)
Protocol
Protocol
Take 1 of the water sample bottles (containing Amount1 L of surface water) and carefully transfer water into a sterile 2 L Pyrex bottle.

If needed, add control strain to water sample by gently tipping vial over the opening. Close bottle tightly and thoroughly mix sample.
Mix
Aseptically measure Amount250 mL of 5X UPB and pour into 2 L bottle (making final concentration of 1X UPB)

Secure lid and mix thoroughly before incubating at Temperature37 °C for 18-24 h.
Note
For next steps, refer to “Selective Enrichment Protocol for Salmonella Isolation from Surface Water.”


Incubation
Mix