![]() vulnificus enumeration, the standard FDA approved protocol will be used. Similarly, 100 microl of homogenate will be plated onto thiosulfate Citrate Bile-Salt and Sucrose (TCBS) agar in triplicate for total Vibrio plate count. One hundred microl sample will be plated onto MA in triplicate for total aerobic plate count. Ten-fold dilutions from the homogenate will be made with PBS. The homogenate will be diluted 1:1 (wt/wt) in phosphate-buffered saline (PBS) and homogenized for another 30s. Oyster meat sample and the shell liquid from ten oysters will be pooled together and homogenized. Oysters were washed and scrubbed in tap water with sterile brushes, shucked and weighed in sterile equipment to avoid cross-contamination. Ten oysters will be randomly selected before and during depuration once a day for bacteriological analysis. Bacteriological analysis will include determination of total heterotrophic aerobic plate counts, total Vibrio counts, and V. Salinity, temperature, and dissolved oxygen will be determined everyday. ![]() During depuration, oysters will be fed twice a day with marine microalgae concentrate (Shellfish diet 1800, Reedmariculture, Campbell, CA). Cleaned oysters will be treated under UV filtered seawater system using a water flow-through system for 7 days at the Auburn University Shellfish Laboratory (AUSL). Oysters will be scrubbed with a sterile scrub-brush under running tap water to remove extraneous material. Basically, the general depuration protocols will be as follows: 120 live shellstock eastern oysters will be acclimated in 19 mm mesh size Durethane oyster bags, (ADPI Enterprise Inc., Philadelphia, PA) for 2 weeks in filtered water flow-through system before depuration. Project Methods Depuration experiments will be based on previous studies performed in our laboratory.
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