title: Procedure for isolation and enumeration of Vibrio parahaemolyticus.
abstract: An evaluation of criteria used in the identification of Vibrio parahaemolyticus showed that cultural responses varied with respect to growth in broth with 10% NaCl, type of hemolysis, reactions in triple sugar-iron-agar, and serological reactions. With few or no exceptions, cultures were positive for cytochrome oxidase, utilized glucose fermentatively, were sensitive to pteridine (0/129) and novobiocin, and failed to grow in Trypticase soy broth (TSB) without NaCl. A procedure employing a direct plating technique, with or without prior enrichment, was designed for the isolation and enumeration of V. parahaemolyticus. The plating medium consisted of 2.0% peptone, 0.2% yeast extract, 1.0% corn starch, 7% NaCl, and 1.5% agar, with the pH adjusted to 8.0. The enrichment broth was TSB with 7% NaCl. Dilutions of food homogenates were either spread directly on the plates or inoculated into enrichment broth. TSB enrichments were incubated at 42 C for 18 hr. A loopful of the TSB tubes then was streaked onto the direct plating medium. Incubation of plates was at 42 C for 24 to 48 hr. Smooth, white to creamy, circular, amylase-positive colonies were then picked as suspect V. parahaemolyticus. Confirmation of gram-negative, fermentative, oxidase-positive, pleomorphic rods sensitive to pteridine 0/129 was made by a fluorescent-antibody technique. With this procedure, a satisfactory quantitative recovery of known V. parahaemolyticus from inoculated seafoods was made possible. V. parahaemolyticus was nto isolated from other salted foods.
mesh_terms: D000681:Amylases / Q000096:biosynthesis; D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis; D001431:Bacteriological Techniques; D003470:Culture Media; D004352:Drug Resistance, Microbial; D003576:Electron Transport Complex IV / Q000096:biosynthesis; D004912:Erythrocytes / Q000276:immunology; D005285:Fermentation; D005396:Fish Products; D005455:Fluorescent Antibody Technique; D005516:Food Microbiology; D005947:Glucose / Q000378:metabolism; D006461:Hemolysis; D006801:Humans; D007106:Immune Sera; D009675:Novobiocin / Q000494:pharmacology; D011621:Pteridines / Q000494:pharmacology; D011817:Rabbits; D012756:Sheep; D014733:Vibrio / Q000145:classification / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D000942:Antigens, Bacterial; D003470:Culture Media; D007106:Immune Sera; D011621:Pteridines; D009675:Novobiocin; D003576:Electron Transport Complex IV; D000681:Amylases; D005947:Glucose