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Background The 3M™ Petrifilm™ Coliform Count (CC) Plate is a sample-ready-culture medium system which contains modified Violet Red Bile nutrients, a cold-water-soluble gelling agent, and a tetrazolium indicator that facilitates colony enumeration. Objective To validate the 3M Petrifilm CC Plate method for bottled water through the AOAC® Performance Tested Method(s)SM program. Methods The performance of the 3M Petrifilm CC Plate was compared to the U.S. Food and Drug Administration Bacteriological Analytical Manual Chapter 4, Section III.D for the enumeration of coliforms in bottled water. Matrix data were normalized by log10 transformation and performance indicators included repeatability and difference of means with 90 and 95% confidence intervals. Inclusivity, exclusivity, robustness, and product consistency and stability were evaluated. Results This study demonstrated that the 3M Petrifilm CC Plate method detects and enumerates coliforms from bottled water. The average log10 counts of the 3M Petrifilm CC Plate method were equivalent to or better than the average log10 counts of the reference method. Results from inclusivity and exclusivity studies demonstrated that the 3M Petrifilm CC Plate method differentiated coliforms from non-coliform strains. In product consistency, stability, and robustness testing, different lots of 3M Petrifilm CC Plates and small deviations in incubation time and temperature did not affect test results. Conclusion The 3M Petrifilm CC Plate method is an accurate and robust method for the enumeration of coliforms in bottled water. Highlights The 3M Petrifilm CC Plate allows for detection of confirmed coliforms within 24 h. Up to 20 sample-ready plates can be stacked during incubation, providing flexibility and saving space.

A validation study of the 3M™ Petrifilm™ Environmental Listeria (EL) Plate (3M Food Safety, St. Paul, MN) was conducted at Q Laboratories, Inc., Cincinnati, OH. The method was compared to the Health Canada MFHPB-30 reference method for the analysis of stainless steel environmental surfaces. Twenty replicates of the environmental surface were analyzed at a low and a high inoculum level. The low-level sampling area was inoculated with 0.2–2 CFU/5 cm2, and the high-level sampling area was inoculated with 2–5 CFU/5 cm2. Five control replicates were also analyzed at 0 CFU/5 cm2. There was no significant difference in the number of positives detected by the 3M Petrifilm EL Plate method and the Health Canada MFHPB-30 reference method for the environmental surface analyzed in this study.

With the rapid growth of the dairy industry and the establishment of strict antimicrobial residue limits in the People's Republic of China's (PRC) milk supply, a -lactamase product known as antimicrobial destroyer was introduced into dairy production without regulatory review. We developed a method for detecting this product in milk samples based on a modified cylinder plate method. The presence of -lactamase is defined as a difference between the inhibitory zones of the test samples (supplemented with 25 g/mL sulbactam plus 0.5 g/mL penicillin G) and control samples (supplemented only with 0.5 g/mL penicillin G) 3 mm. Using this method, 77 individually packaged milk samples were randomly collected from 5 retail stores in 3 cities over a 4-month period (May to August 2006). Of the 77 samples, 49 were found to be -lactamase-positive. In 2 undiluted milk samples showing extremely high -lactamase activity, 25 g/mL sulbactam could not inhibit penicillin G activity. Because there is a lack of safety data on -lactamases in milk products, these data indicated a potentially serious safety concern for the dairy industry in the PRC.

A new method for detection of Salmonella in foods in 48 h has been granted AOAC First Action approval in selected foods (Official Method 2001.08) using both the VIDAS Immuno-Concentration Salmonella (ICS) method and a combination of 3 selective plates: Hektoen enteric (HE), bismuth sulfite (BS), and xylose lysine desoxycholate (XLD).

The 3M™ Petrifilm™ Staph Express Count plate method was compared with AOAC Official Method 975.55 for the enumeration of Staphylococcus aureus in selected foods. Four foods—cooked, diced chicken; cured ham; smoked salmon; and pepperoni—were analyzed for S. aureus by 12 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample, a low inoculation level, a medium inoculation level, and a medium inoculation level with background flora, each in duplicate. The mean log10 counts for the methods were comparable for all 4 foods. The repeatability and reproducibility variances of the 24 h Petrifilm Staph Express Count plate method were similar to those of the 72 h standard method.

The 3M™ Petrifilm™ Staph Express Count plate method was compared with AOAC Official Method 975.55 for the enumeration of Staphylococcus aureus in selected foods. Five foods—ice cream, raw milk, yogurt, whey powder, and cheese—were analyzed for S. aureus by 12 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample, a low inoculation level, a medium inoculation level, and a medium inoculation level with background flora, each in duplicate. The mean log10 counts for the methods were comparable for all 5 foods. The repeatability and reproducibility variances of the 24 h Petrifilm Staph Express Count plate method were similar to those of the 72 h standard method.

The practice of detecting and enumerating all oxidase-negative, glucose-fermenting-Gram-negative rods (i.e., the family Enterobacteriaceae) isused to indicate unsanitary or inadequate food processing conditions. The objective of this interlaboratory collaborative study was to evaluate and compare the methods described in Standard Methods for the Examination of Dairy Products (SMEDP) and the Compendium of Methods for the Microbiological Examination of Foods (Compendium) with a commercial product, the 3M™ Petrifilm™ Enterobacteriaceae Count Plate, for the recovery of Enterobacteriaceae in foods. Six foods—cheddar cheese, milk, flour, frozen prepared meals, frozen broccoli, and nut pieces—were analyzed for Enterobacteriaceae by 12 collaborating laboratories. For each food tested, the collaborators received 8 blind test portions consisting of a control test portion and 3 levels of inoculated test portion, each in duplicate. Each test portion was tested by the Petrifilm Enterobacteriaceae Count Plate method as well as the SMEDP or Compendium methods. The precision estimates (repeat-ability or within-laboratory variation, and reproducibility or between-laboratory variation) were calculated with standard statistical techniques.

The VIDAS Immuno-concentration Salmonella (ICS) plus selective plate method (Hektoen enteric, xylose lysine desoxycholate, bismuth sulfite) method for the detection of Salmonella was compared to the Bacteriological Analytical Manual (BAM)/AOAC culture method in a collaborative study. Thirty-two laboratories participated in the evaluation. Each laboratory tested one or more of the 6 test products: milk chocolate, nonfat dry milk, dried whole egg, soy flour, ground black pepper, and ground raw turkey. The 2 methods were in agreement for 1297 of the 1455 samples. Of the 158 samples not in agreement, 82 were VIDAS ICS plus selective plate-positive and BAM/AOAC-negative, and 76 were VIDAS ICS plus selective plate-negative and BAM/AOAC-positive.

The VIDAS Immuno-concentration Salmonella (ICS) plus selective plate method (Hektoen enteric, bismuth sulfite, Salmonella identification) method for the detection of Salmonella was compared to the Bacteriological Analytical Manual (BAM)/AOAC culture method in a collaborative study. Thirty-two laboratories participated in the evaluation. Each laboratory tested one or more of the 6 test products: milk chocolate, nonfat dry milk, dried whole egg, soy flour, ground black pepper, and ground raw turkey. The 2 methods were in agreement for 1283 of the 1440 test samples. Of the 157 test samples not in agreement, 82 were VIDAS ICS plus selective plate-positive and BAM/AOAC-negative, and 75 were VIDAS ICS plus selective plate-negative and BAM/AOAC-positive.

A collaborative study with Enterobacteriaceae was conducted to validate Hygicult® E dipslides by comparison with violet red bile glucose agar (VRBGA) contact plates and swabbing, using stainless steel surfaces artificially contaminated with microbes at various levels. Twelve laboratories participated in the validation procedure. The total number of collaborative samples was 108. The microbial level in each sample was assessed in triplicate by using the 3 above-mentioned methods. No Enterobacteriaceae were used at the low inoculation level. At the middle inoculation level, the percentages detached from the test surfaces were 16.6 with the Hygicult E method, 15.3 with the contact plate method, and 14.6 with swabbing; at the high innoculation level, the percentages were 14.5, 15.8, and 9.8, respectively. The percentage of acceptable results after the removal of outliers was 97.2. Repeatability relative standard deviations ranged from 33.4 to 44.9%; reproducibility relative standard deviations ranged from 45.2 to 77.1%. The Hygicult E dipslide, VRBGA contact plate, and swabbing methods gave similar results at all 3 microbial levels tested

A rehydratable dry-film plating method for coliforms in foods, the 3M™ Petrifilm™ Rapid Coliform Count plate method, was compared with the U.S. Food and Drug Administration's Bacteriological Analytical Manual method for nondairy foods and the American Public Health Association's Standard Methods for the Examination of Dairy Products (SMEDP) method for dairy foods. Six food types, vanilla ice cream, cheddar cheese, fresh refrigerated uncooked pasta, wheat flour, prepared frozen macaroni and cheese, and frozen hash browns, were analyzed for coliforms by 11 collaborating laboratories. For each food product tested, the collaborators received 8 blind samples consisting of a control sample and 3 levels of inoculated sample, each in duplicate. The mean log counts for the methods were comparable. The repeatability and reproducibility variances of the Petrifilm Rapid Coliform Count method at 14 and 24 h were not significantly different from those of the standard methods.

A rehydratable dry-film plating method for Staphylococcus aureusin foods, the 3M™ Petrifilm™ Rapid S. aureus Count Plate method, was compared with AOAC® Official MethodSM 975.55 (Staphylococcus aureus in Foods). Nine foods—instant nonfat dried milk, dry seasoned vegetable coating, frozen hash browns, frozen cooked chicken patty, frozen ground raw pork, shredded cheddar cheese, fresh green beans, pasta filled with beef and cheese, and egg custard—were analyzed for S. aureus by 13 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample and 3 levels of inoculated test sample, each in duplicate. The mean log counts for the methods were comparable for pasta filled with beef and cheese; frozen hash browns; cooked chicken patty; egg custard; frozen ground raw pork; and instant nonfat dried milk. The repeatability and reproducibility variances of the Petrifilm Rapid S. aureus Count Plate method were similar to those of the standard method.

Results with the new 3M™ Petrifilm™ Rapid S. aureus Count (RSA) Plate method were compared with those of the classical Baird-Parker agar (BPA) method for detection and enumeration of Staphylococcus aureus. Studies on 219 bacterial strains demonstrated that the Petrifilm RSA plate is more sensitive than and as specific as the classical BPA method for confirmed identification of S. aureus. Counts of colonies from 71 pure cultures, 61 naturally contaminated food samples, and more than 750 artificially inoculated food samples showed that the Petrifilm RSA method was as effective as the classical BPA method for identification and enumeration of S. aureus. The Petrifilm RSA method gave results in one-third the time required for the classical method.

A rehydratable dry-film plating method for Escherichia coli, the Petrifilm E. coli/Coliform (EC) Count Plate in foods, has been compared with the AOAC INTERNATIONAL most probable number (MPN) method. Eleven laboratories participated in the collaborative study. Three E. coli levels in 8 samples each of frozen raw ground turkey, frozen raw ground beef, and frozen cooked fish were tested in duplicate. Mean log counts for the Petri film plate procedure were not significantly different from those for the MPN procedure for cooked fish samples inoculated with low or high inocula levels, for samples of raw turkey inoculated at medium level, and for beef inoculated at low, medium, and high levels. Repeatability and reproducibility vari ances of the Petrifilm EC Plate method recorded at 24 h were as good as or better than those of the MPN method. The dry rehydratable film method for enumerating confirmed E. coli in poultry, meats, and seafood has been adopted first action by AOAC INTERNATIONAL.

Made available in DSpace on 2022-04-29T08:47:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-01-01 The validation of a microbiological assay, applying cylinder plate method for determination of the activity of lomefloxacin in coated tablets is described. Using a strain of Bacillus subtilis ATCC 9372 as the test organism, lomefloxacin was measured in concentrations ranging from 2.0 to 8.0 μg/mL. The method validation showed that it is linear (r= 0.9999), precise (relative standard deviation = 1.15%), and accurate (it measured the added quantities). The excipients did not interfere in the determination. It was concluded that the microbiological assay is satisfactory for quantitation of lomefloxacin in tablets. Universidade Estadual Paulista (UNESP) Faculdade de Ciências Farmacêuticas UNESP, Rod. Araraquara-Jaú, km 1, CEP 14801-902, Araraquara, SP Universidade Estadual Paulista (UNESP) Faculdade de Ciências Farmacêuticas UNESP, Rod. Araraquara-Jaú, km 1, CEP 14801-902, Araraquara, SP

A test procedure for evaluating the effect of adding commercial liquid hand dishwashing detergents to kitchen sponges to control microbial growth is described. Claims for this type of application are being made on dishwashing detergents throughout the world. In this evaluation, commercially available kitchen sponges were stripped of antimicrobial compounds. Sponges were then inoculated with a pool of 7 microorganisms which consisted of Gram positives, Gram negatives, and yeast. Inoculated sponges were treated with the detergent as recommended by the manufacturer and allowed to incubate for 16 h at ambient temperature. Surviving microorganisms were then quantitated using either the spiral or pour plate method. Tests were run using both clean sponges and sponges soiled with 0.5% nonfat dry milk (NFDM). Untreated sponges showed stasis or slightly increased bacterial populations after the incubation period in the absence of NFDM. Significant increases of up to 3 log cfu/mL were observed for untreated sponges when soiled with NFDM. Statistically significant reductions were observed for clean sponges (99.8–99.9998%) and sponges soiled with NFDM (87.6–99.9%) when detergents making “antibacterial sponge” claims were added to the inoculated sponges. Statistically significant differences between detergents making “antibacterial sponge” claims were also observed.

In previous experiments, the turbidimetric method for determining chlortetracycline-HCI (CTC-HCl) in feeds showed lower recovery than the AOAC plate method. Although the addition of vitamins to the turbidimetric medium improved results, values by the turbidimetric method remained about 10% lower than by the plate method. A modified (1.7 × the weight recommended by the manufacturer) turbidimetric assay medium decreased assay sensitivity but did not significantly change the slope of S. aureus response to CTC-HCl. There was no evidence that vitamin fortification of the modified medium had any significant effect on the growth rate of test organism. Examination of about 100 samples of commercial feeds containing CTC-HCl showed excellent agreement in results between the turbidimetric and plate methods.

Assay of bacitracin activity in feeds is subject to many sources of variation. Bacitracin standards are complex mixtures and may contain various amounts of less active forms of bacitracin. Test organisms may respond differently to these mixtures. Collaborative studies should involve the use of one standard, perhaps USP zinc bacitracin. The use of different types of bacitracin standards (regular or zinc) in a laboratory may contribute to variation in sample potency. The pyridine extraction method is subject to serious operational difficulties. Among the contributing factors are incomplete evaporation of pyridine (causing positive bias), allowing the temperature to increase too much during evaporation (causing negative bias), and the numerous manipulative steps in the technique (causing large variability). Because of these factors and the toxic properties of pyridine, the method finds little use. Methanol extraction offers a better substitute for determining zinc or methylene disalicylate bacitracin in premixes and complete feeds by the plate method.

The manual and automated turbidimetric assays and a modified official plate assay for Chlortetracycline (CTC-HCl) in feed were collaboratively studied. Three feed samples (swine feed, 100 g CTC-HCl/ton; premix I, 20 g each of CTC-HCl and sulfamethazine/lb, and 10 g penicillin/lb; and premix II, 50 g CTC-HCl/lb) were analyzed at 2 dilutions. Twelve laboratories conducted the plate assay; 8 laboratories, the manual turbidimetric method ; and 7 laboratories, the Autoturb analysis. Within a method, there was no significant difference between dilutions. Between methods, there was a significant difference between the manual turbidimetric plate assays only for swine feed. However, the same sample dilutions or the average values of the 2 dilutions for both methods showed no statistical difference. Among the collaborators, the slope of CTC-HCl standard curve varied between about 2.0 and 3.0 for the plate method. The turbidimetric assay has been adopted as official first action for feeds containing >20 g CTC-HCl/lb.

The spiral plate count method is a semiautomated plating technique that greatly reduces manpower and material costs normally associated with the pour plating technique. In this collaborative study, 8 laboratories compared the spiral and pour plating techniques, using 4 samples each of 3 products : frozen pumpkin pie, frozen chicken pot pie, and shampoo. The results show that 10 of the 12 comparisons of means of the pour and spiral methods were not significantly different; 2 values were significant at α = 0.01. Overall, the components of variance were less than that of the current milk standard, and the replicate per cent coefficient of variation was satisfactory. This study indicates that the spiral plate method is an acceptable alternative to the pour plate method; the spiral plate method has been adopted as official first action.

Two laboratories participated in a comparison of a manual turbidimetric assay with the AUTOTURB® System and the AOAC method of analysis of tylosin in 4 feed samples. Results showed no significant difference between the 2 turbidimetric assays. When the AOAC method was considered, the difference between laboratories was significant. On an overall basis the turbidimetric methods were significantly higher than the plate method. The relative standard deviation was higher (6.72%) for the plate assay versus turbidimetric assay (4.5%). The mean recovery in both laboratories was significantly less than the labeled amount of tylosin by all 3 methods.