Iron Milk Medium Method for Recovering Clostridium perfringens from Shellfish: Collaborative Study

1994 ◽  
Vol 77 (2) ◽  
pp. 351-356 ◽  
Author(s):  
Carlos Abeyta ◽  
June H Wetherington

Abstract Eleven laboratories participated in a collaborative study analyzing shellfish (oysters, Crassostrea gigas) for the detection and enumeration of Clostridium perfringens by the iron milk medium (IMM) method. The IMM method was compared to AOAC Official Method 976.30. Shellfish were artificially inoculated with C. perfringens cells (vegetative and spores) at low (1 × 103 colony forming units [cfu]/g), medium (1 × 104 cfu/g), and high (1 × 106 cfu/g) levels. Negative controls (zero level) were analyzed by each laboratory. C. perfringens FD-1, the strain involved in a foodborne illness, was used. Blind duplicates of each inoculum level were analyzed, giving a total of 16 samples per laboratory. The selectivity of IMM relies solely on the rapid growth of C. perfringens at 45°C indicated by stormy fermentation reaction within 18 h. C. perfringens is detected and enumerated using the most probable number technique. A statistical evaluation of the data found no significant differences between the estimates from the 2 methods. The IMM method for detection of C. perfringens from shellfish has been adopted first action by AOAC INTERNATIONAL.

2006 ◽  
Vol 89 (1) ◽  
pp. 115-126 ◽  
Author(s):  
Hidemasa Kodaka ◽  
Hajime Teramura ◽  
Tadanobu Nirazuka ◽  
Shingo Mizuochi ◽  
David Goins ◽  
...  

Abstract Compact Dry CF is a ready-to-use test method for the enumeration of coliform bacteria in food. The plates are presterilized and contain culture medium and a cold water-soluble gelling agent. The medium should be rehydrated with 1 mL diluted sample inoculated into the center of the self-diffusible medium, allowing the solution to diffuse by capillary action. The plate can be incubated at 35C for 2024 h and the colonies counted without any further working steps. The Compact Dry CF medium plates were validated with 5 different raw meats. The performance tests were conducted at 35C. In all studies performed, no apparent differences were observed between the Compact Dry CF method and the AOAC Official Method 966.24 results. For the accuracy claim (n = 75), a correlation factor of r2 = 0.91 (coliform) could be assigned, as stated in the application for Performance-Tested MethodSM. No significant variations in coliform bacterial counts were observed with different production lots or plates of diverse storage age by the quality consistency and storage robustness studies.


2003 ◽  
Vol 66 (1) ◽  
pp. 120-121
Author(s):  
JOHN KAROLUS ◽  
MERCURIA CUMBO ◽  
SUSAN BOEHLER ◽  
LAURA SAVINA

Four hundred fifteen shellfish seawater samples from approved, conditionally approved, and restricted areas along the coastlines of Connecticut, Massachusetts, and Maine were tested in duplicate to compare results obtained with A-1 medium (AOAC official method no. 978.23) and those obtained with A-1 medium without salicin. Four laboratories used five sets of most probable number procedures to perform the analyses. No statistically significant differences between the two media were found with the t test, the F test, or the analysis of variance.


1997 ◽  
Vol 80 (6) ◽  
pp. 1235-1271 ◽  
Author(s):  
Jane E Fox Dobson ◽  
Foster D McClure ◽  
Alvin P Rainosek ◽  
K Dashiell ◽  
J Fox Dobson ◽  
...  

Abstract An intralaboratory1collaborative study evaluated a modified version of AOAC Official Method 971.13 for determining the fish flesh content (FFC) in frozen coated fish products by comparing it with the on-line method. Eleven collaborators analyzed 36 products (a total of 6336 test samples). Each product targeted one of 4 percent fish flesh (PFF) levels (35,50,65, and 80). Products were manufactured from one of 3 raw materials (fillet blocks, minced blocks, and natural fillets) and processed in one of 4 forms (sticks, portions, formed portions, and fillets) and one of 4 styles (raw breaded, batter-dipped, precooked, and fully cooked). Each “official” test sample was tracked through the processing system and weighed (1) before battering and/or breading and, depending on product style, before frying; and (2) after battering and/or breading and, depending on product style, after frying; so that it served as its own control.


1996 ◽  
Vol 79 (4) ◽  
pp. 907-916 ◽  
Author(s):  
Joanna M Lynch ◽  
David M Barbano ◽  
J Richard Fleming

Abstract A modified Mojonnier ether extraction method for determination of the fat content of cream was developed based on the method for milk (AOAC Official Method 989.05). The cream Babcock method (AOAC Official Method 920.111 B-C) was modified to harmonize with the milk Babcock method (AOAC Official Method 989.04) and to clarify procedural details. Using the AOAC collaborative study format, 10 laboratories tested 9 pairs of blind duplicate heat-treated cream samples with a fat range of 30-45% using both methods. The statistical performance (invalid and outlier data removed) was as follows: mean % fat = 37.932, sr = 0.125, sR = 0.151, RSDr = 0.330, RSDR = 0.398, r = 0.354, and R = 0.427 for the ether extraction method. For the Babcock method, mean % fat = 38.209, sr = 0.209, SR = 0.272, RSDr = 0.548, RSDR = 0.712, r = 0.592, and R = 0.769. Average test results for fat from the Babcock method were 0.277% (absolute fat) greater than for the Mojonnier ether extraction method. The difference between methods, as a percentage of the average fat content of the samples, was 0.73%. This agrees with differences observed between the 2 methods for milk when 10 to 17 laboratories tested 7 milk samples in blind duplicate at bimonthly intervals over a 4-year period (average difference 0.029% fat, 0.78% as a percentage of average fat content). The Mojonnier ether extraction and Babcock methods for fat in cream have been adopted by AOAC INTERNATIONAL. The new Babcock method replaced the AOAC Official Method 920.111 B-C.


1982 ◽  
Vol 65 (5) ◽  
pp. 1129-1133 ◽  
Author(s):  
William D St John ◽  
Jack R Matches ◽  
Marleen M Wekell

Abstract A simple iron milk medium was used for isolation and enumeration of Clostridium perfringens from soil, sludge, and water samples. The whole milk contained only iron powder as a reducing agent; no other inhibitors were added. The iron milk most probable number (MPN) procedure was compared with 4 plating media: sulfite-polymyxin-sulfadiazine, Shahidi-Ferguson perfringens, tryptose-sulfite- cycloserine (both with and without egg yolk), and tryptone-sulfite-neomycin. The selectivity of the iron milk relies solely on the rapid growth of C. perfringens at 45°C and the stormy fermentation reaction within 18 h. Isolates were confirmed as C. perfringens by standard biochemical tests. The iron milk MPN procedure compared very well with the 4 plating media tested. Selectivity of incubation temperature, short incubation time, and ease of identification by the characteristic stormy fermentation make this method ideal for enumerating C. perfringens from large numbers of samples.


2005 ◽  
Vol 68 (2) ◽  
pp. 331-335 ◽  
Author(s):  
V. NAVARRO-HIDALGO ◽  
E. CABRERA-DÍAZ ◽  
H. ZEPEDA ◽  
L. MOTA DE LA GARZA ◽  
A. CASTILLO ◽  
...  

A quantitative survey of Clostridium perfringens in typical foods served at local restaurants was conducted for 18 months in Guadalajara, Mexico. A total of 151 samples, including goat's birria (50), pozole (50), and beef tamales (51), were collected from small restaurants in Guadalajara. Samples were tested for C. perfringens by the most probable number (MPN) method and for mesophilic aerobic plate counts (MAPCs) and coliform, yeast, and mold counts by plate count methods. Isolates confirmed as C. perfringens were further sporulated and tested for cytotoxic or cytotonic effect against Vero cells as an indication of enterotoxin production. C. perfringens was detected in 78 (52%) of all samples at concentrations that ranged from 2.3 to 5.4 log MPN/g. Average MAPCs were 1.3 to 2.7 log CFU/g, depending on the type of dish. Coliform counts ranged from less than 1.0 to 1.5 CFU/g, and yeast and mold counts were less than 1.0 log CFU/g in all cases. A total of 118 isolates of C. perfringens were tested for enterotoxic effect on Vero cells; 82 (70%) showed activity against Vero cells. Of them, 31 isolates induced cell lysis, indicating cytotoxic effect; 41 induced cell elongation, indicating cytotonic effect; and 10 produced both cytotoxic and cytotonic effect. Dilution of the bacterial filtrates that were still producing an effect on Vero cells ranged from 1:80 to 1:5,120. These results underscore the importance of determining enterotoxigenicity when testing for C. perfringens in foods.


1987 ◽  
Vol 70 (1) ◽  
pp. 35-38 ◽  
Author(s):  
Gayle A Lancette ◽  
John Lanier

Abstract Enumeration of Staphylococcus aureus in foods was collaboratively studied by comparing the present AOAC final action method, 46.062, which uses trypticase soy broth with 10% NaCl to a proposed replacment method which uses the same broth with 1% sodium pyruvate added. Fifteen collaborators analyzed uninoculated samples of milk, tuna salad, and ground turkey, as well as samples inoculated with low (102 cells/g), middle (104 cells/g), and high (106 cells/g) levels of S. aureus. The samples were frozen immediately to maintain the inoculated level of S. aureus in the food. A different strain of S. aureus was used for each food; heat-stressed S. aureus cells were used to inoculate the milk samples. The pyruvate-amended broth significantly (α = 0.05) increased enumeration of low, middle, and high levels of S. aureus from milk and ground turkey, and from tuna salad at middle and high levels. The pyruvate-amended media method has been adopted official first action to replace method 46.062


2013 ◽  
Vol 96 (1) ◽  
pp. 86-90 ◽  
Author(s):  
Larry M Brooks ◽  
Benjamin J Kuhlman Kuhlman ◽  
Doug W McKesson ◽  
Leo McCloskey

Abstract The poor interoperability of anthocyanin glycosides measurements by two pH differential methods is documented. Adams-Harbertson, which was proposed for commercial winemaking, was compared to AOAC Official MethodSM 2005.02 for wine. California bottled wines (Pinot Noir, Merlot, and Cabernet Sauvignon) were assayed in a collaborative study (n = 105), which found mean precision of Adams- Harbertson winery versus reference measurements to be 77 ± 20%. Maximum error is expected to be 48% for Pinot Noir, 42% for Merlot, and 34% for Cabernet Sauvignon from reproducibility RSD. Range of measurements was actually 30 to 91% for Pinot Noir. An interoperability study (n = 30) found Adams-Harbertson produces measurements that are nominally 150% of the AOAC pH differential method. Large analytical chemistry differences are: AOAC method uses Beer-Lambert equation and measures absorbance at pH 1.0 and 4.5, proposed a priori by Flueki and Francis; whereas Adams-Harbertson uses “universal” standard curve and measures absorbance ad hoc at pH 1.8 and 4.9 to reduce the effects of so-called co-pigmentation. Errors relative to AOAC are produced by Adams-Harbertson standard curve over Beer-Lambert and pH 1.8 over pH 1.0. The study recommends using AOAC Official Method 2005.02 for analysis of wine anthocyanin glycosides.


2006 ◽  
Vol 89 (1) ◽  
pp. 100-114 ◽  
Author(s):  
Hidemasa Kodaka ◽  
Shingo Mizuochi ◽  
Hajime Teramura ◽  
Tadanobu Nirazuka ◽  
David Goins ◽  
...  

Abstract Compact Dry E. coli/Coliform Count (EC) is a ready-to-use test method for the enumeration of Escherichia coli and coliform bacteria in food. The plates are presterilized and contain culture medium and a cold water-soluble gelling agent. The medium should be rehydrated with 1 mL diluted sample inoculated onto the center of the self-diffusible medium, allowing the solution to diffuse by capillary action. The plate can be incubated at 35C for 2024 h and the colonies counted without any further working steps. The Compact Dry EC medium plates were validated as an analysis tool for determining colony-forming units (CFU) of E. coli and coliform bacteria from a variety of raw meats using 5 different types of raw meats. The performance tests were conducted at 35C. In all studies performed, no apparent differences were observed between the Compact Dry ECmethod and theAOAC Official Method 966.24 results. For the accuracy claim (n = 75), a correlation factor of r2 = 0.93 (E. coli) and r2 = 0.93 (coliform bacteria) could be assigned, as stated in the application for Performance-Tested MethodSM.


2003 ◽  
Vol 86 (5) ◽  
pp. 963-970 ◽  
Author(s):  
Karen M Silbernagel ◽  
Robert P Jechorek ◽  
Charles N Carver ◽  
Barbara L Horter ◽  
Kathryn G Lindberg ◽  
...  

Abstract 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.


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