Evaluating survival of Escherichia coli O157:H7 in frozen and thawed apple cider: potential use of a hydrophobic grid membrane filter-SD-39 agar method. 1998

J R Sage, and S C Ingham
University of Wisconsin-Madison, Department of Food Science 53706-1565, USA.

To determine the susceptibility of Escherichia coli O157:H7 to freezing and thawing in apple cider, methods that recover injured cells are needed for accurate enumeration. This study compared the ISO-GRID hydrophobic grid membrane filter (HGMF) SD-39 agar method to two other methods: a reference most probable number (MPN) method, and plating on sorbitol MacConkey agar (SMA). To determine numbers of injured cells, SMA spread plating was also compared to Trypticase soy agar (TSA) spread plating. Two strains of E. coli O157:H7 QA 326 and ATCC 43895, were inoculated into presterilized apple cider (10 ml) which was then frozen (-20 degrees C for 24 h). Samples were thawed at 4 degrees C for 4 h, or at 23 degrees C for 1.5 h, or in a microwave oven (700 W for 10 s). Substantial cell death (0.69- to 6.33 log10 CFU/ml decreases) and injury (0.70- to 2.38-log10 CFU/ml decreases) occurred during freezing and thawing. The extent of death and injury varied with strain and thawing method. The TSA spread plating method recovered the most cells while the HGMF method always recovered more viable cells than the reference MPN method and also either recovered significantly more (P < 0.05) cells or a not significantly different number of cells than SMA spread plating. Some injured cells of both strains were not counted by the HGMF method. Significant numbers of cells injured by freezing and thawing at 4 degrees C in apple cider were enumerated in the cider was diluted 1:2 Trypticase soy broth immediately before plating. Two epifluorescent microscopic methods showed that injury was not associated with loss of cell membrane integrity.

UI MeSH Term Description Entries
D008567 Membranes, Artificial Artificially produced membranes, such as semipermeable membranes used in artificial kidney dialysis (RENAL DIALYSIS), monomolecular and bimolecular membranes used as models to simulate biological CELL MEMBRANES. These membranes are also used in the process of GUIDED TISSUE REGENERATION. Artificial Membranes,Artificial Membrane,Membrane, Artificial
D003470 Culture Media Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN. Media, Culture
D005374 Filtration A process of separating particulate matter from a fluid, such as air or a liquid, by passing the fluid carrier through a medium that will not pass the particulates. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Filtrations
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
D000362 Agar A complex sulfated polymer of galactose units, extracted from Gelidium cartilagineum, Gracilaria confervoides, and related red algae. It is used as a gel in the preparation of solid culture media for microorganisms, as a bulk laxative, in making emulsions, and as a supporting medium for immunodiffusion and immunoelectrophoresis.
D001431 Bacteriological Techniques Techniques used in studying bacteria. Bacteriologic Technic,Bacteriologic Technics,Bacteriologic Techniques,Bacteriological Technique,Technic, Bacteriological,Technics, Bacteriological,Technique, Bacteriological,Techniques, Bacteriological,Bacteriologic Technique,Bacteriological Technic,Bacteriological Technics,Technic, Bacteriologic,Technics, Bacteriologic,Technique, Bacteriologic,Techniques, Bacteriologic
D001628 Beverages Liquids that are suitable for drinking. (From Merriam Webster Collegiate Dictionary, 10th ed) Beverage
D015169 Colony Count, Microbial Enumeration by direct count of viable, isolated bacterial, archaeal, or fungal CELLS or SPORES capable of growth on solid CULTURE MEDIA. The method is used routinely by environmental microbiologists for quantifying organisms in AIR; FOOD; and WATER; by clinicians for measuring patients' microbial load; and in antimicrobial drug testing. Agar Dilution Count,Colony-Forming Units Assay, Microbial,Fungal Count,Pour Plate Count,Spore Count,Spread Plate Count,Streak Plate Count,Colony Forming Units Assay, Microbial,Colony Forming Units Assays, Microbial,Agar Dilution Counts,Colony Counts, Microbial,Count, Agar Dilution,Count, Fungal,Count, Microbial Colony,Count, Pour Plate,Count, Spore,Count, Spread Plate,Count, Streak Plate,Counts, Agar Dilution,Counts, Fungal,Counts, Microbial Colony,Counts, Pour Plate,Counts, Spore,Counts, Spread Plate,Counts, Streak Plate,Dilution Count, Agar,Dilution Counts, Agar,Fungal Counts,Microbial Colony Count,Microbial Colony Counts,Pour Plate Counts,Spore Counts,Spread Plate Counts,Streak Plate Counts
D019453 Escherichia coli O157 A verocytotoxin-producing serogroup belonging to the O subfamily of Escherichia coli which has been shown to cause severe food-borne disease. A strain from this serogroup, serotype H7, which produces SHIGA TOXINS, has been linked to human disease outbreaks resulting from contamination of foods by E. coli O157 from bovine origin. E coli O157,E coli O157-H7,Escherichia coli O157-H7
D019685 Rosales An order of the class MAGNOLIOPSIDA. Its members include some of the most known ornamental and edible plants of temperate zones including roses, apples, cherries, and peaches. Pittosporaceae,Pittosporum,Pittosporums,Rosale

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