The influence of adaptation to pH (from pH 5.0 to 9.0) on membrane lipid composition, verotoxin concentration, and resistance to acidic conditions in simulated gastric fluid (SGF) (pH 1.5, 37°C) was determined for Escherichia coli O157:H7 (HEC, ATCC 43895), an rpoS-deficient mutant of ATCC 43895 (HEC-RM, FRIK 816-3), and nonpathogenic E. coli (NPEC, ATCC 25922). Regardless of the strain, D values (in SGF) of acid-adapted cells were higher than those of non-acid-adapted cells, with HEC adapted at pH 5.0 having the greatest D value, i.e., 25.6 min. Acid adaptation increased the amounts of palmitic acid (C16:0) and decreased cis-vaccenic acid (C18:17c) in the membrane lipids of all strains. The ratio of cis-vaccenic acid to palmitic acid increased at acidic pH, causing a decrease in membrane fluidity. HEC adapted to pH 8.3 and HEC-RM adapted to pH 7.3 exhibited the greatest verotoxin concentrations (2,470 and 1,460 ng/ml, respectively) at approximately 10 8 CFU/ml. In addition, the ratio of extracellular to intracellular verotoxin concentration decreased at acidic pH, possibly due to the decrease of membrane fluidity. These results suggest that while the rpoS gene does not influence acid resistance in acid-adapted cells it does confer decreased membrane fluidity, which may increase acid resistance and decrease verotoxin secretion.
Oysters at the retail stage of distribution generally contain greater densities of Vibrio parahaemolyticus than do oysters at harvest. The objective of this study was to determine the effects of postharvest storage at 26 and 3 degrees C on the growth and survival of naturally occurring V. parahaemolyticus in shellstock American oysters (Crassostrea virginica). Oysters were collected monthly from May 1998 through April 1999 from Mobile Bay, Alabama, and their V. parahaemolyticus densities were determined after 0, 5, 10, and 24 h of postharvest storage at 26 degrees C. After 24 h of storage at 26 degrees C, oysters were transferred to a refrigerator at 3 degrees C and analyzed 14 to 17 days later. V. parahaemolyticus numbers were determined by a direct plating method involving an alkaline-phosphatase-labeled DNA probe that targets the species-specific thermolabile hemolysin gene (tlh-AP) to identify suspect isolates. From April to December, when water temperatures at harvest were >20 degrees C, the geometric mean harvest density of V. parahaemolyticus was 130 CFU/g. When water temperatures were <20 degrees C, the geometric mean harvest density was 15 CFU/g. After harvest, V. parahaemolyticus multiplied rapidly in live oysters held at 26 degrees C, showing a 50-fold increase (1.7 log CFU/g) at 10 h and a 790-fold increase (2.9 log CFU/g) at 24 h (April through December). Average V. parahaemolyticus numbers showed a sixfold decrease (0.8 log CFU/g) after approximately 14 days of refrigeration. These results indicate that V. parahaemolyticus can grow rapidly in unrefrigerated oysters.
The occurrence of antibiotic-resistant bacteria in foods of animal origin is a potential health threat because resistance can be transferred among bacteria, and antibiotic-resistant pathogens may not respond to antibiotic treatments. Thirteen brands of ready-to-eat shrimp representing four countries of origin were obtained from local grocery stores. Total heterotrophic plate counts were determined, and antibiotic-resistant bacteria were isolated. Total heterotrophic colony counts ranged from 3.3 to 5.6 log CFU/g, which was within approved quality limits. A total of 1,564 isolates representing 162 bacterial species were recovered during screening of resistance to 10 antibiotics: ampicillin, ceftriaxone, chloramphenicol, clindamycin, erythromycin, nalidixic acid, streptomycin, tetracycline, trimethoprim, and vancomycin. Six hundred fifty-seven (42%) of the isolates and 131 (81%) of the species had acquired resistance to antibiotics. Numerous resistant human pathogens were isolated, including Escherichia coli, Enterococcus spp., Salmonella, Shigella flexneri, Staphylococcus spp., and Vibrio spp. Nonresistant Yersinia spp. also were isolated. Ready-to-eat shrimp is sold with instructions to thaw the product before serving, which may result in consumer exposure to antibiotic-resistant bacteria. Widespread trade of this product provides an avenue for international dissemination of antibiotic-resistant pathogens.
Changes in total volatile base nitrogen (TVBN), pH, salt soluble protein (SSP), moisture content, and expressible moisture of channel catfish frame mince during storage at Ϫ20, 0, and 5°C were investigated. Refined mince was either unwashed or washed twice, with mince designated for frozen storage mixed with cryoprotectant. TVBN increased during refrigerated storage, while SSP decreased during frozen storage. Expressible moisture increased during frozen storage but not during refrigeration storage temperatures. Moisture content and pH of mince did not change during storage. Results indicate that mince should be stored for no longer than 3 d at 0 or 5°C to maintain optimal quality. Frozen mince with cryoprotectant would remain acceptable for at least 3 months at Ϫ20°C.
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