Full scare commercial pasteurization equipment operated at 72-73°C with a holding time of 15-16 s was used to determine the ability of commercial thermal processing to inactivate Listeria monocytogenes strain Scott A. Three methods of providing L. monocytogenes concentration in raw milk were employed: freely suspended (extra-cellular), inside bovine phagocytes (in vitro procedure), and inside bovine phagocytes in experimentally infected cows (in vivo). Three enrichment methods were used to assay for L. monocytogenes after pasteurization: cold enrichment (4°C, 28 d), selective enrichment of Lovett et al. (FDA procedure) (17), and the USDA-FSIS two stage enrichment procedure. In addition, a 1-L sample taken just before the vacuum breaker was incubated undiluted in the original sample container (4°C, 4 weeks). None of the four assay methods could detect Listeria in the pasteurized milk.

The lethality of Listeria isolates was determined with normal adult mice and mice that were immunocompromised by treatment with 20 mg of carrageenan per kg. The mean 50% lethal doses (LD50s) of the pathogenic isolates were significantly lower (alpha = 0.05) in the immunocompromised mice than in the untreated mice, with an average reduction of 5.8 log10 units. In contrast, the mean LD50s of the nonpathogenic isolates were lower in the immunocompromised mice by an average of only 0.4 log10 unit, a difference that was not significant (alpha = 0.05). When immunocompromised mice were used, the LD50s of pathogenic Listeria monocytogenes isolates were lower than those of nonpathogenic L. innocua and L. seeligeri isolates by greater than or equal to 6 log10 units and lower than those of nonpathogenic L. ivanovii isolates by greater than or equal to 4 log10 units. Pathogenic L. monocytogenes isolates could be distinguished from nonpathogenic isolates by their ability to cause deaths in immunocompromised mice in 3 days at a dose of approximately 10(4) CFU per mouse. An alternative procedure using iron-overloaded mice failed to effectively differentiate pathogenic Listeria isolates.

The method of Doyle and Roman (Appl. Environ. Microbiol. 43:1343-1353, 1982) was compared with that of Lovett et al. (Appl. Environ. Microbiol. 46:459-462, 1983) for the ability to recover Campylobacter jejuni strains inoculated into raw milk at a concentration of less than 1 cell per g. The method of Lovett et al. gave significantly greater recovery proportions.

To establish an enrichment system of high efficiency for recovery of Campylobacter jejuni from market chickens, the effects of the temperature, duration of incubation, and pH of the enrichment culture on the isolation of the bacterium were evaluated. Whole chickens or chicken parts in plastic bags were individually rinsed, and the washings filtered through cheesecloth. The cells were separated from the washings by centrifugation, and the pellet was inoculated into 100 mL of enrichment broth. Isolation of C. jejuni from poultry samples was significantly increased by incubating these samples in an enrichment medium at 42 degrees C as opposed to 35 degrees C; for 48 h as opposed to 24 h or 72 h; and at pH 7.0 as opposed to pH 6.0, 6.5, 7.5, or 8.0.

Campylobacter jejuni was isolated from raw milk by a method that can routinely detect less than or equal to 1 organism per ml. This procedure was used in a survey of 195 separate farms and showed a 1.5% incidence of C. jejuni in milk from bulk tanks.

Of 790 samples of oyster shellstock freshly harvested during a 12-month survey, 111 (most of which were harvested from June through August) contained Vibrio cholerae non-O1 (611 strains), and seven contained O1 Inaba (11 strains) organisms. None of the V. cholerae strains isolated were enterotoxigenic by immunological and biological tests.

Three of 36 raw milk isolates of Yersinia enterocolitica produced enterotoxin in milk at 25 degrees C, but not at 4 degrees C. No strain tested could survive pasteurization.