The Presence of Norovirus and Adenovirus on Environmental Surfaces in Relation to the Hygienic Level in Food Service Operations Associated with a Suspected Gastroenteritis Outbreak

Erratum: Food and environmental virology 9 (2017):3, pp. 358-359 DOI.10.1007/s12560-017-9304-6Norovirus (NoV) gastroenteritis outbreaks appear frequently in food service operations (FSOs), such as in restaurants and canteens. In this study the presence of NoV and adenovirus (AdV) genomes was investigated on the surfaces of premises, especially in kitchens, of 30 FSOs where foodborne gastroenteritis outbreaks were suspected. The objective was to establish a possible association between the presence of virus genomes on surfaces and a visual hygienic status of the FSOs. NoV genome was found in 11 and AdV genome in 8 out of 30 FSOs. In total, 291 swabs were taken, of which 8.9% contained NoV and 5.8% AdV genome. The presence of NoV genomes on the surfaces was not found to associate with lower hygiene level of the premises when based on visual inspection; most (7/9) of the FSOs with NoV contamination on surfaces and a completed evaluation form had a good hygiene level (the best category). Restaurants had a significantly lower proportion of NoV-positive swabs compared to other FSOs (canteens, cafeteria, schools etc.) taken together (p = 0.00014). The presence of a designated break room for the workers was found to be significantly more common in AdV-negative kitchens (p = 0.046). Our findings suggest that swabbing is necessary for revealing viral contamination of surfaces and emphasis of hygiene inspections should be on the food handling procedures, and the education of food workers on virus transmission.Peer reviewe

Significance of official food control in food safety : food business operators' perceptions

Finnish food business operators' (FBOs) opinions of the food control inspections performed by local authorities were evaluated, using a questionnaire. The production types of FBOs included were slaughterhouses, meat, fish and milk plants, egg-packing plants and storage facilities dealing with foods of animal origin. Based on a total of 459 responses, we noted that the impacts of official controls were considered valuable for food safety, since 78.8% of the respondents saw that the actions taken based on inspections had enhanced the safety of the products. The results also highlighted the importance of inspectors being familiar with the production processes to increase the efficacy of food control. More frequent visits by official inspectors correlated positively with FBOs' conceptions of noncompliances being relevant for food safety. The FBOs saw local inspectors as the most important sources of new information concerning food safety legislation and 89.1% of the respondents confirmed that discussions with local inspectors had helped them understand the food safety risks within their processes. We also noted that the bigger the FBO, the more clearly they seem to perceive the risks associated with their processes (Spearman's rank correlation coefficient r = 0.127, P = 0.009)

Efficient DNA fingerprinting of Clostridium botulinum types A, B, E, and F by amplified fragment length polymorphism analysis.

Amplified fragment length polymorphism (AFLP) analysis was applied to characterize 33 group I and 37 group II Clostridium botulinum strains. Four restriction enzyme and 30 primer combinations were screened to tailor the AFLP technique for optimal characterization of C. botulinum. The enzyme combination HindIII and HpyCH4IV, with primers having one selective nucleotide apiece (Hind-C and Hpy-A), was selected. AFLP clearly differentiated between C. botulinum groups I and II; group-specific clusters showed <10% similarity between proteolytic and nonproteolytic C. botulinum strains. In addition, group-specific fragments were detected in both groups. All strains studied were typeable by AFLP, and a total of 42 AFLP types were identified. Extensive diversity was observed among strains of C. botulinum type E, whereas group I had lower genetic biodiversity. These results indicate that AFLP is a fast, highly discriminating, and reproducible DNA fingerprinting method with excellent typeability, which, in addition to its suitability for typing at strain level, can be used for C. botulinum group identification

Elimination of botulinum neurotoxin (BoNT) type B from drinking water by small-scale (personal-use) water purification devices and detection of BoNT in water samples.

Seven small-scale drinking water purification devices were evaluated for their capacity to eliminate botulinum neurotoxin (BoNT) type B from drinking water. Influent water inoculated with toxic Clostridium botulinum cultures and effluent purified water samples were tested for the presence of BoNT by using a standard mouse bioassay and two commercial rapid enzyme immunoassays (EIAs). The water purification devices based on filtration through ceramic or membrane filters with a pore size of 0.2 to 0.4 µm or irradiation from a low-pressure UV-lamp (254 nm) failed to remove BoNT from raw water (reduction of 2.3 log10 units). The rapid EIAs intended for the detection of BoNT from various types of samples failed to detect BoNT from aqueous samples containing an estimated concentration of BoNT of 396,000 ng/liter

Diversity of proteolytic Clostridium botulinum strains, determined by a pulsed-field gel electrophoresis approach

Pulsed-field gel electrophoresis (PFGE) was applied to the study of the similarity of 55 strains of proteolytic Clostridium botulinum (C. botulinum group I) types A, AB, B, and F. Rare-cutting restriction enzymes ApaI, AscI, MluI, NruI, PmeI, RsrII, SacII, SmaI, and XhoI were tested for their suitability for the cleavage of DNA of five proteolytic C. botulinum strains. Of these enzymes, SacII, followed by SmaI and XhoI, produced the most convenient number of fragments for genetic typing and were selected for analysis of the 55 strains. The proteolytic C. botulinum species was found to be heterogeneous. In the majority of cases, PFGE enabled discrimination between individual strains of proteolytic C. botulinum types A and B. The different toxin types were discriminated at an 86% similarity level with both SacII and SmaI and at an 83% similarity level with XhoI. Despite the high heterogeneity, three clusters at a 95% similarity level consisting of more than three strains of different origin were noted. The strains of types A and B showed higher diversity than the type F organisms which formed a single cluster. According to this survey, PFGE is to be considered a useful tool for molecular epidemiological analysis of proteolytic C. botulinum types A and B. However, epidemiological conclusions based on PFGE data only should be made with discretion, since highly similar PFGE patterns were noticed, especially within the type B strains

Antibacterial efficiency of Finnish spice essential oils against pathogenic and spoilage bacteria

Reprinted with permission from the Journal of Food Protection. Copyright held by the International Association for Food Protection, Des Moines, Iowa, U.S.A

Thermal inactivation of non-proteolytic Clostridium botulinum type E spores in model fish media and in vacuum-packaged hot-smoked fish products.

Thermal inactivation of nonproteolytic Clostridium botulinum type E spores was investigated in rainbow trout and whitefish media at 75 to 93°C. Lysozyme was applied in the recovery of spores, yielding biphasic thermal destruction curves. Approximately 0.1% of the spores were permeable to lysozyme, showing an increased measured heat resistance. Decimal reduction times for the heat-resistant spore fraction in rainbow trout medium were 255, 98, and 4.2 min at 75, 85, and 93°C, respectively, and those in whitefish medium were 55 and 7.1 min at 81 and 90°C, respectively. The z values were 10.4°C in trout medium and 10.1°C in whitefish medium. Commercial hot-smoking processes employed in five Finnish fish-smoking companies provided reduction in the numbers of spores of nonproteolytic C. botulinum of less than 103. An inoculated-pack study revealed that a time-temperature combination of 42 min at 85°C (fish surface temperature) with >70% relative humidity (RH) prevented growth from 106 spores in vacuum-packaged hot-smoked rainbow trout fillets and whole whitefish stored for 5 weeks at 8°C. In Finland it is recommended that hot-smoked fish be stored at or below 3°C, further extending product safety. However, heating whitefish for 44 min at 85°C with 10% RH resulted in growth and toxicity in 5 weeks at 8°C. Moist heat thus enhanced spore thermal inactivation and is essential to an effective process. The sensory qualities of safely processed and more lightly processed whitefish were similar, while differences between the sensory qualities of safely processed and lightly processes rainbow trout were observed

Multiplex PCR assay for detection and identification of Clostridium botulinum types A, B, E and F in food and fecal material.

Botulism is diagnosed by detecting botulinum neurotoxin and Clostridium botulinum cells in the patient and in suspected food samples. In this study, a multiplex PCR assay for the detection of Clostridium botulinum types A, B, E, and F in food and fecal material was developed. The method employs four new primer pairs with equal melting temperatures, each being specific to botulinum neurotoxin gene type A, B, E, or F, and enables a simultaneous detection of the four serotypes. A total of 43 C. botulinum strains and 18 strains of other bacterial species were tested. DNA amplification fragments of 782 bp for C. botulinum type A alone, 205 bp for type B alone, 389 bp for type E alone, and 543 bp for type F alone were obtained. Other bacterial species, including C. sporogenes and the nontoxigenic nonproteolytic C. botulinum-like organisms, did not yield a PCR product. Sensitivity of the PCR for types A, E, and F was 102 cells and for type B was 10 cells per reaction mixture. With a two-step enrichment, the detection limit in food and fecal samples varied from 102 spore/g for types A, B, and F to 101 spore/g of sample material for type E. Of 72 natural food samples investigated, two were shown to contain C. botulinum type A, two contained type B, and one contained type E. The assay is sensitive and specific and provides a marked improvement in the PCR diagnostics of C. botulinum

Sensitivity study of the instrumental temperature corrections on Brewer total ozone column measurements

The instrumental temperature corrections to be applied to the ozone measurements by the Brewer spectrophotometers are derived from the irradiance measurements of internal halogen lamps in the instruments. These characterizations of the Brewer spectrophotometers can be carried out within a thermal chamber, varying the temperature from -5 to +45ºC, or during field measurements, making use of the natural change in ambient temperature. However, the internal light source used to determine the thermal sensitivity of the instrument could be affected in both methods by the temperature variations as well, which may affect the determination of the temperature coefficients. In order to validate the standard procedures for determining Brewer’s temperature coefficients, two independent experiments using both external light sources and the internal halogen lamps have been performed within the ATMOZ Project. The results clearly show that the traditional methodology based on the internal halogen lamps is not sensitive to the temperature-caused changes in the spectrum of the internal light source. The three methodologies yielded equivalents results, with differences in total ozone column below 0.08% for a mean diurnal temperature variation of 10ºC.This work has been supported by the European Metrology Research Programme (EMRP) within the joint research project ENV59 “Traceability for atmospheric total column ozone” (ATMOZ)