Comparison of the Petrifilm dry rehydratable film and conventional culture methods for enumeration of yeasts and molds in foods: collaborative study.

A collaborative study was performed involving 18 laboratories and 6 food types to compare 3M Petrifilm yeast and mold count plates with the method described in the U.S. Food and Drug Administration's Bacteriological Analytical Manual. Four species of mold and 2 species of yeast were used to inoculate the following foods: hot dogs, corn meal, ketchup, orange juice, yogurt, and cake mix. Each collaborator received 15 samples of each food type: 5 low-level inoculations, 5 high-level inoculations, and 5 uninoculated samples. There was no significant difference between the means of the 2 methods for any product or inoculation level. The Petrifilm yeast and mold count plate method for enumeration of yeasts and molds in foods has been adopted first action by AOAC INTERNATIONAL.

High-sensitivity dry rehydratable film method for enumeration of coliforms in dairy products: collaborative study.

A dry-film coliform count plate that is inoculated with 5 mL sample was compared with the Violet Red Bile Agar plate method in a collaborative study by 18 laboratories. Products analyzed were 2% milk, chocolate milk, cream, vanilla ice cream, cottage cheese, and cheese. Collaborators tested blind duplicate uninoculated samples and samples inoculated at low, medium, and high level. Significantly (P < 0.05) higher numbers of coliforms were recovered by the dry-film method from 2% milk samples at the 3 inoculum levels, the chocolate milk at the low- and high-inoculum levels, and the cream at the high-inoculum level. Significantly higher counts were obtained by the agar method for cottage cheese samples at the low-inoculum level. The repeatability standard deviation for the dry-film method was significantly higher for the high-inoculum level chocolate milk sample and the medium-inoculum level cottage cheese. The same statistic was significantly higher for the agar method at all 3 inoculum levels in the 2% milk and the medium-inoculum level cream. The high-sensitivity dry rehydratable film method for enumeration of coliforms in dairy products has been adopted first action by AOAC INTERNATIONAL.

Dry rehydratable film for enumeration of total coliforms and Escherichia coli in foods: collaborative study.

Rehydratable dry-film plating methods for total coliforms and Escherichia coli in foods have been compared to the AOAC most probable number methods. Fourteen laboratories participated in the collaborative study. Three coliform and E. coli levels in 6 samples of 4 product types (flour, nuts, cheese, and beef with gravy) and in 3 samples of 2 product types (mushrooms and raw turkey) were tested in duplicate by the participants. The mean log counts for the 3 methods were comparable. In general, the repeatability and reproducibility variances of the plating methods were as good as or better than that of the MPN method. The method has been adopted official first action by AOAC.

Dry rehydratable film for enumeration of total aerobic bacteria in foods: collaborative study.

A rehydratable dry-film plating procedure for aerobic plate counts has been compared to the standard agar plate method (966.23B and C, 15th ed.; 46.014-46.015, 14th ed.) in a collaborative study by 12 laboratories. Each laboratory analyzed the normal microflora of 3 samples in duplicate for 6 products. The aerobic plate counts ranged from 1.0 x 10(3) to 1.0 x 10(8) cfu/g. The products were flour, nuts, frozen raw shrimp, spice, frozen raw ground turkey, and frozen and refrigerated vegetables. Repeatability standard deviations of the 2 methods did not differ significantly for 13 of 18 test samples. For 1 shrimp and 2 turkey samples, the dry-film method had lower repeatability variances (P less than 0.05) and for 1 spice sample the agar method had lower repeatability variances (P less than 0.05). Relative standard deviations of repeatability were between 1.7 and 15.5% for the dry-film method and 1.2 and 16.0% for the agar method. Relative standard deviations of reproducibility ranged from 2.4 to 23.4% for the dry-film method and 2.3 to 18.8% for the agar method. The dry rehydratable film method has been adopted official first action for determination of the aerobic plate count.

Dry rehydratable films for enumeration of coliforms and aerobic bacteria in dairy products: collaborative study.

A collaborative study was conducted to compare proposed dry-film plating methods, using aerobic count plates and coliform count plates, to standard agar plating methods for quantifying aerobic bacteria and coliforms in dairy products. In this study, 5 food products (chocolate milk, pasteurized cheese, nonfat dry milk, evaporated milk, and vanilla ice cream), selected as representative dairy products, were analyzed by 11 collaborating laboratories. The results indicate that the dry-film plating methods are equivalent to or better than the agar plating methods. The aerobic count and coliform count dry-film plating methods have been adopted official first action.

Evaluation of the 3M Petrifilm™ Culture Plate Method for Enumerating Aerobic Flora and Coliforms in Poultry Processing Facilities.

Petrifilm™ methods were compared to traditional plating methods for monitoring microbial contamination in poultry processing facilities. No differences were seen between the Petrifilm methods and conventional method for enumeration of total bacterial populations, and no trends were seen in the ability of either method to detect coliforms in naturally contaminated samples. When processed poultry products were artificially inoculated with a variety of microorganisms, no difference in efficiency of recovery of the bacteria was noted. The Petrifilm methods were shown to be a practical and accurate alternative for monitoring microbial levels in poultry processing facilities.

Microbial colonization of the intestinal epithelium in suckling mice.

Colonization by indigenous microorganisms of the mucosal epithelia of the large bowels of suckling mice was followed by microbial culture techniques and by light, fluorescence, and electron microscopy. Certain microbes colonize in distinctive patterns the cecal and colonic epithelia in these mice. Coliforms and enterococci colonize the large bowel 7 to 9 days after birth and reach high population levels during the second week. During that period, these facultative anaerobes can be detected by immunofluorescence techniques in microcolonies in the mucin on the epithelium. During the third week, however, after their populations decline to the low levels characteristic of adult mice, coliforms and enteroccoci can be observed only infrequently in the mucous layer. Anaerobic fusiform-shaped bacteria appear in the mucous layers along with the microcolonies of enterococci and coliforms during the second week after birth. These anaerobes increase in numbers in the mucin until they form thick layers on the mucosal epithelium by the end of the third week. They remain in the mucous layer throughout the life of the normal mouse. Anaerobic spiral-shaped microbes also colonize the mucous layer on the cecal and colonic epithelium. But these organisms can be detected by immunofluorescence in 1-week-old mice, well in advance of the time the fusiform-shaped bacteria can be found. In the second week, the latter microbes co-inhabit the mucous layer with the spiral-shaped organisms. The fusiform- and spiral-shaped microbes remain associated in the mucin on the cecal and colonic mucosal epithelia into the adult life of mice.

Anaerobic bacteria on the mucosal epithelium of the murine large bowel.

Anaerobic bacteria can be detected at population levels of 10(11) organisms per g of cecum or colon in adult mice from four different colonies widely spaced in the United States. Most of these microorganisms are oxygen-intolerant fusiform-shaped bacteria. At least one type of these tapered, rod-shaped bacteria can be seen in layers in the epithelial mucin in frozen-section histological preparations of the large bowels of mice. In addition, such microorganisms can be seen within 0.5 mum of the epithelium in ultrathin sections of colon or cecum examined in an electron microscope. These fusiform-shaped bacteria predominate in the mucin layers. However, spiral-shaped microorganisms can be found as well near the mucosal epithelia in ultrathin sections of colon. Also, such organisms can be seen in negatively-stained preparations of washings of the colonic mucosal epithelia examined in an electron microscope. At least three types of spiral-shaped organisms, including both spiral-shaped bacteria and spirochetes, can be found in preparations from mice from three of the four colonies. Such spiral-shaped microorganisms can be detected at population levels as great as 10(9) organisms per g of cecum or colon in anaerobic cultures of the large bowels of mice from all four colonies. One anaerobic spiral bacterium was isolated in pure culture. This particular organism was found by immunofluorescence to be intermingled with the fusiform-shaped bacteria in the mucin on the mucosal epithelium in the mouse large bowel.

Cecal enlargement and microbial flora in suckling mice given antibacterial drugs.

Enlargement and microbial colonization of the cecum were examined in neonatal mice suckling mothers drinking either water or an aqueous solution of penicillin. The full ceca increased in weight at the same rate in both drug-treated and control mice during the first 15 to 17 days after birth. Thereafter, cecal weight increased at a greater rate in the drug-treated animals than in the untreated controls. At weaning, the ceca in treated mice were two to three times the size of control organs and remained enlarged as long as penicillin was given. The enlarged ceca did not differ histologically from those in controls. From birth, the cecal microflora in the drug-treated mice differed qualitatively and quantitatively and in colonization pattern from the flora of control mice. The ceca of untreated animals were colonized primarily by large populations of lactobacilli during the first week after birth, small populations of coliforms and enterococci during the second week, and enormous populations of bacteroides and certain gram-negative fusiform-shaped anaerobic bacteria during the third week. In contrast, the organs of the treated mice were populated by large populations of coliforms and enterococci during the first week and enormous populations of clostridia and unusual gram-negative nonsporeforming bacteria during the third week. These large abnormal populations were present in the ceca as they enlarged during the third week after birth in the drug-treated animals. These findings confirm that only certain populations of anaerobic bacteria can act to maintain cecal size in normal animals.