Effect of temperature on the survival of F-specific RNA coliphage, feline calicivirus, and Escherichia coli in chlorinated water.

We compared the survival of F-specific RNA coliphage MS2, feline calicivirus, and E. coli in normal tap water and in tap water treated to an initial concentration of 50 ppm free chlorine and held at 4 degrees C, 25 degrees C, or 37 degrees C for up to 28 days. Our aim was to determine which of these two organisms (coliphage or E. coli) was better at indicating norovirus survival under the conditions of the experiment. There was a relatively rapid decline of FCV and E. coli in 50 ppm chlorine treated water and both organisms were undetectable within one day irrespective of the temperature. In contrast, FRNA phage survived for 7 to 14 days in 50 ppm chlorine treated water at all temperatures. All organisms survived for 28 days in tap water at 4 degrees C, but FCV was undetectable on day 21 and day 7 at 25 degrees C and 37 degrees C, respectively. Greater survival of FRNA phage compared to E. coli in 50 ppm chlorine treated water suggests that these organisms should be further investigated as indicators of norovirus in depurated shellfish, sanitized produce, and treated wastewater which are all subject to high-level chlorine treatment.

Occurrence of Escherichia coli, noroviruses, and F-specific coliphages in fresh market-ready produce.

Forty samples of fresh produce collected from retail food establishments were examined to determine the occurrence of Escherichia coli, F-specific coliphages, and noroviruses. An additional six samples were collected from a restaurant undergoing investigation for a norovirus outbreak. Nineteen (48%) of the retail samples and all outbreak samples were preprocessed (cut, shredded, chopped, or peeled) at or before the point of purchase. Reverse transcription-PCR, with the use of primers JV 12 and JV 13, failed to detect norovirus RNA in any of the samples. All six outbreak samples and 13 (33%) retail samples were positive for F-specific coliphages (odds ratio undefined, P = 0.003). Processed retail samples appeared more likely to contain F-specific coliphages than unprocessed samples (odds ratio 3.8; 95% confidence interval 0.8 to 20.0). Only two (5.0%) retail samples were positive for E. coli; outbreak samples were not tested for E. coli. The results of this preliminary survey suggest that F-specific coliphages could be useful conservative indicators of fecal contamination of produce and its associated virological risks. Large-scale surveys should be conducted to confirm these findings.

Effect of temperature and sanitizers on the survival of feline calicivirus, Escherichia coli, and F-specific coliphage MS2 on leafy salad vegetables.

We conducted a series of experiments to compare the survival of Escherichia coli, feline calicivirus, and F-specific coliphage MS2 on lettuce and cabbage with and without disinfection. Inoculated produce was held at 4, 25, or 37 degrees C for 21 days or was treated with different concentrations of sodium bicarbonate, chlorine bleach, peroxyacetic acid, or hydrogen peroxide. Survival was measured by the decimal reduction value (time to 90% reduction in titer) and the change in log titers of the test organisms. A stronger correlation of survival measures was observed between feline calicivirus and MS2 than between E. coli and either of the viral agents at 25 and 37 degrees C. The maximum time to detection limit for MS2 at all temperatures was 9 days, whereas feline calicivirus was detected for a maximum of 14 days at 4 degrees C. In contrast, E. coli was detectable for 21 days at 4 and 25 degrees C and for 14 days at 37 degrees C. Significant increases in E. coli titer occurred within the first 5 days, but virus titers decreased steadily throughout the experiments. E. coli was also highly susceptible to all disinfectants except 1% sodium bicarbonate and 50 ppm chlorine bleach, whereas the viruses were resistant to all four disinfectants.

Hand washing compliance among retail food establishment workers in Minnesota.

Inadequate hand washing by food workers is an important contributing factor to foodborne disease outbreaks in retail food establishments (RFEs). We conducted a survey of RFEs to investigate the effect of hand washing training, availability of hand washing facilities, and the ability of the person in charge (PIC) to describe hand washing according to the Minnesota Food Code (food code) on workers' ability to demonstrate food code-compliant hand washing. Only 52% of the PICs could describe the hand washing procedure outlined in the food code, and only 48% of workers could demonstrate code-compliant hand washing. The most common problems observed were failure to wash for 20 s and failure to use a fingernail brush. There was a strong positive association between the PIC being a certified food manager and being able to describe the food code hand washing procedure (odds ratio [OR], 5.5; 95% confidence interval [CI], 2.2 to 13.7), and there was an even stronger association between the PIC being able to describe hand washing and workers being able to demonstrate code-compliant hand washing (OR, 15; 95% CI, 6 to 37). Significant associations were detected among correct hand washing demonstration, physical infrastructure for hand washing, and the hand washing training methods used by the establishment. However, the principal determinant of successful hand washing demonstration was the PIC's ability to describe proper hand washing procedure. These results suggest that improving hand washing practices among food workers will require interventions that address PIC knowledge of hand washing requirement and procedure and the development and implementation of effective hand washing training methods.

Survival of F-specific RNA coliphage, feline calicivirus, and Escherichia coli in water: a comparative study.

The relationship between the survival of enteric viral pathogens and their indicators (coliform bacteria and coliphages) is not well understood. We compared the survival rates of feline calicivirus (FCV), Escherichia coli, and a male-specific RNA coliphage MS2 at 4, 25, and 37 degrees C for up to 28 days in dechlorinated water. The survival rates of E. coli and FCV, a surrogate of noroviruses (NV), had a high degree of correlation at 4 and 25 degrees C, while MS2 phage survived significantly longer (P < 0.05) at these two temperatures. At 37 degrees C, the survival rates for all three organisms were highly correlated. Decimal reduction values indicating the number of days needed for 90% reduction in titer (D values) decreased for all three organisms as storage temperatures increased. FCV had the shortest D value among all three organisms at all temperatures investigated. These findings indicate that F-specific RNA phages may be useful indicators of NV in the environment.

Concentration and detection of caliciviruses from food contact surfaces.

Outbreaks of human Norwalk virus (NV) and Norwalk-like viruses often originate in food service establishments. No reliable method is available for the detection of these human caliciviruses on food contact surfaces. We describe a simple method for the detection of NV from stainless steel work surfaces using cultivable feline calicivirus (FCV) as a model. Stainless steel surfaces were artificially contaminated with known amounts of FCV, followed by its elution in a buffer solution. Three methods of virus elution were compared. In the first method, moistened cotton swabs or pieces of nylon filter (1MDS) were used to elute the contaminating virus. The second method consisted of flooding the contaminated surface with eluting buffer, allowing it to stay in contact for 15 min, followed by aspiration of the buffer (aspiration method) after a contact period of 15 min. The third method, the scraping-aspiration method, was similar to the aspiration method, except that the surfaces were scraped with a cell scraper before buffer aspiration. Maximum virus recovery (32 to 71%) was obtained with the scraping-aspiration method using 0.05 M glycine buffer at pH 6.5. Two methods (organic flocculation and filter adsorption elution) were compared to reduce the volume of the eluate recovered from larger surfaces. The organic flocculation method gave an average overall recovery of 55% compared to the filter-adsorption-elution method, which yielded an average recovery of only 8%. The newly developed method was validated for the detection of NV by artificial contamination of 929-cm2 stainless steel sheets with NV-positive stool samples and for the detection of the recovered virus by reverse transcription-polymerase chain reaction.