D.

Analysis

17.3.04
AOAC Official Method 991.14
Coliform and Escherichia coli
Counts in Foods

See 989.10B(b)–(f) (see 17.3.03).
E. coli count plates.—Plates are similar to coliform count plates,
989.10B(b) (see 17.3.03), with addition of5-bromo-4-chloro-3-indolyl-β-Dglucuronide, an indicator of glucuronidase activity. This allows coliforms
and E. coli to be read on the same plate. Petrifilm E. coli Count Plates (Microbiology Products, 3M Center, Bldg. 275-5W-05, St. Paul, MN 55144, USA)
meet these specifications.

(a) Coliform count.—Place dry-film E. coli count plate, B, or
coliform count plate, 989.10B(b) (see 17.3.03), on flat surface. Lift
top film and inoculate 1 mL test suspension onto center of film base.
Carefully place top film down onto inoculum. Distribute test suspension over prescribed growth area with downward pressure on center
of plastic spreader device (flat side down). Leave plate undisturbed
1 min to permit gel to solidify. Incubate plates 24 ± 2 h at 35° ± 1°C.
In incubator, place plates in horizontal position, clear side up, in
stacks not exceeding 20 units. Count plates promptly after incubation period. After incubation is complete, plates may be stored
frozen (≤–15°C) up to 7 days. This should be avoided as routine
practice. Use standard colony counter for counting purposes. Magnifier-illuminator may also be used to facilitate counting. Coliforms appear as red colonies that have one or more gas bubbles associated
(within one colony diameter) with them. Count all colonies in countable
range (15–150 colonies). Red colonies without gas bubbles are not
counted as coliform organisms.
(b) E. coli count.—Use E. coli count plate and proceed as in (a). Incubate an additional 24 ± 2 h (48 ± 4 h total). E. coli colonies appear as
blue colonies associated with gas bubbles; other coliforms appear as red
colonies with gas.

C. Preparation of Test Suspension

Reference: JAOAC 74, 635(1991).

Dry Rehydratable Film
(Petrifilmä E. coli/Coliform Count Plateä and
Petrifilmä Coliform Count Plateä) Methods
First Action 1991
Final Action 1994

See Table 991.14 for the results of the interlaboratory study supporting the acceptance of the method.
A. Principle

See 989.10A (see 17.3.03).
B. Apparatus and Reagent

See 966.23B (see 17.2.01) and 986.33C (see 17.3.02).

Revised: March 2002

© 2002 AOAC INTERNATIONAL

285 2.9 19.549 0.120 1.109 3.268 2.2 3.4 7.1 Low Medium High Low Medium High Low Medium High Low Medium High Low Medium High Low Medium High 1.4 6.436 0.7 21.835 0.243 2.316 0.320 0.7 63.106 0.001 0.318 1.112 0.028 0.1 21.8 16.744 0.233 0.746 4.732 0.291 1.3 2.4 9.261 0.6 12.215 2.345 0.9 3.7 13.699 0.605 0.154 0.5 36.9 13.069 0.1 16.9 Low Medium High Low Medium High Low Medium High Low Medium High Low Medium High Low Medium High 1.550 0.192 0.6 62.896 0.3 12.0 15.728 0.296 0.716 1.058 3.929 0.3 18.098 0.074 2.2 9.141 0.8 60.2 6.759 0.753 0.857 22.8 27.722 1.257 2.9 3. % Total coliforms on coliform count plates Ground turkey Fresh mushrooms Beef with gravy Cheese Wheat flour Nut meal Ground turkey Fresh mushrooms Beef with gravy Cheese Wheat flour Nut meal Ground turkey Fresh mushrooms Beef with gravy Cheese Wheat flour Nut meal Low Medium High Low Medium High Low Medium High Low Medium High Low Medium High Low Medium High 1.227 0.265 © 2002 AOAC INTERNATIONAL .2 24.242 0.6 33.6 9.8 10.2 12.8 6. coli on dry-film count plates Product Level Mean log.216 0.067 0.9 19.8 22.166 0.278 0.883 1.172 2.8 3.316 0.2 4.819 2.8 3.2 37.460 0.6 33.364 0.3 49.4 21.6 13.378 2.105 4.411 0.9 8.355 0.361 0.233 0.7 11.760 0.440 1.3 38.808 1.212 2.737 1.346 1.343 0.331 0.845 0.113 4. coli on E.5 10.0 35.175 3.0 47.172 0.5 52.5 16.422 0.364 0.486 2.049 0.9 14.6 21.0 40.268 0.783 0.296 0.028 1.9 8.662 5.432 0.821 0.524 0.8 6.236 0.3 23.600 5.261 0.500 3.079 5.295 0.0 3.2 2.288 0. coli count plates 0.190 0.639 0.557 0.1 21.9 4.270 0.923 2.6 21.7 37.161 0.0 18.243 3.967 0.8 38.14 Interlaboratory study results for total coliforms and E. % RSDR.4 38.172 0.991 1.5 14.7 26.172 2.873 0.370 0.434 0. cfu/g sr sR RSDr.177 2.062 0.8 51.2 12.364 0.193 2. coli count plates 0.7 15.9 3.235 0.238 0.256 2.5 2.2 11.403 0.8 38.817 0.191 0.6 27.2 8.9 33.123 0.312 0.2 44.095 3.278 4.016 0.0 22.9 18.391 0.1 11.715 0.6 37.4 22.550 0.9 11.092 3.607 2.2 11.6 31.635 0.1 23.620 0.443 0.190 0.3 23.5 15.831 10.7 12.872 0.361 2.5 8.0 0.5 21.601 0.206 0.454 3.1 22.295 0.0 1.267 Total coliforms on E.1 22.3 22.060 0.315 0.3 18.1 4.4 23.Table 991.291 3.7 20.813 0.064 0.314 0.336 1.507 0.4 38.828 0.826 4.442 0.098 0.7 44.3 16.679 0.934 0.284 0.999 0.0 13.1 9.197 0.425 Total E.241 0.874 1.419 1.362 1.1 9.915 9.