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P3-04 Extraction of Enteric Virus Indicator from Seawater Using Activated Carbon

Wednesday, July 25, 2012
Exhibit Hall (Rhode Island Convention Center)
Jiemin Cormier, Louisiana State University, Baton Rouge, LA
Miguel Gutierrez, Louisiana State University, Baton Rouge, LA
Lawrence Goodridge, Colorado State University, Fort Collins, CO
Marlene Janes, Louisiana State University, Baton Rouge, LA
Introduction: Enteric virus-contaminated shellfish represents a significant health threat to shellfish consumers as well as an economic threat to the seafood industry. Male specific bacteriophage MS2 has been identified as a suitable indicator for water-borne enteric virus outbreaks, and its presence indicates a recent contamination. There is an urgent need for the development of rapid detection methods that enable efficient detection of viruses in seawater samples.

Purpose: To investigate if activated carbon can be used for extraction of MS2 bacteriophage from seawater.

Methods: The effects of pH (4, 5, 6, 7, 8, and 9), salinity (0, 10, 20, 30, and 40 ppt) and contact temperature (4 °C, 20 °C and 37 °C) on the absorbance efficiency of activated carbon were investigated in artificial seawater. MS2 bacteriophage at 1E8-1E9 PFU were inoculated into 500 ml of artificial seawater and 1 g of activated carbon was added. After 3 h of constant stirring, activated carbon was separated from the seawater, incubated with 1 ml of trypsin-EDTA solution for 2 h at room temperature to release MS2 bacteriophage from the activated carbon, and RNA was extracted. qRT- PCR was conducted to determine the PFU of MS2 bacteriophage released from the activated carbon.

Results: Results indicated that temperature has a significant effect on the absorbance efficiency of activated carbon. Warmer temperature provides better efficiency for activated carbon. In zero salinity water, the pH has no significant effect on the absorbance efficiency of activated carbon, regardless of the temperature. Activated carbon has significant higher absorbance efficiency in seawater than zero salinity water. Seawater with salinity of 10 ppt at a contact temperature of 37 °C was proven to provide the best absorbance efficiency for activated carbon.

Significance: The use of activated carbon has the potential to be developed into a rapid concentration and detection method for enteric water-borne viruses.

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