Listeria monocytogenes in ready-to-eat (RTE) delicatessen foods: Prevalence, genomic characterization of isolates and growth potential.

This study investigated Listeria monocytogenes prevalence and count in 132 ready-to-eat (RTE) delicatessen samples belonging to different categories (starters with/without mayonnaise pasta/rice-based courses, meat/fish-based main courses) produced by an Italian industry. Whole Genome Sequencing characterized the isolates to map the pathogen circulation. Moreover, the growth potential of L. monocytogenes in the most contaminated product was investigated by a challenge test. L. monocytogenes was detected in 23 samples, giving an estimated prevalence of 17.4 %. Starters with mayonnaise showed a very high prevalence (56.7 %), showing the role of the sauce in the diffusion of the pathogen within the plant. A total of 49 isolates were obtained; they belonged to two different serogroups, IIb and IIa, and were related to two clonal complexes (CCs) and sequence types (STs) (CC288-ST330 and CC121-ST717), suggesting the possible persistence and circulation of the pathogen within the plant. The results of the challenge test showed a limited ability to grow in the selected product thanks to the presence of lactic microflora.

Growth of Listeria monocytogenes in ready-to-eat "shrimp cocktail": Risk assessment and possible preventive interventions.

The present study investigated the presence, growth potential, and public health risk posed by Listeria monocytogenes in a ready-to-eat "shrimp cocktail". The pathogen was detected in 4 out of the 104 samples, and there were no counts above the enumeration limit (1 Log colony-forming unit (CFU)/g); the product was a suitable substrate for pathogen growth owing to its chemical/physical properties. A stochastic quantitative microbial risk assessment (QMRA) was performed to estimate the expected number of invasive listeriosis cases caused by the consumption of 10,000 servings of the product on the last day of its shelf life, considering a population comprising healthy consumers, those susceptible, and transplant recipients. The model predicted no cases for this scenario. Uncertainties were included by considering alternative scenarios; even when considering an increased mean bacterial concentration (up to 3-4 Log CFU/g), no cases were estimated. Following a producer's demand, the exposure assessment model was also used to estimate the probability of the product exceeding the threshold of 2 log CFU/g during the shelf life. The possibility of Listeria growth in the product could not be avoided. Therefore, a modification of the production process was tested to re-classify the product as unsuitable for Listeria growth (EC Reg. 2073/2005). The shrimps were conditioned in three different organic acid solutions comprising: acetic acid (1500 ppm) (A); benzoic acid (1500 ppm) + acetic acid (500 ppm) + lactic acid (750 ppm) (BLA); and lactic acid (4500 ppm) + sodium acetate (2500 ppm) (LSA). Testing was conducted over various treatment durations (1 day-5 days). Treatment for 2 days in the LSA solution was selected based on efficacy, the absence of consumer-perceptible sensorial modifications, and the producers' production rate requirements. The concentration of L. monocytogenes decreased when the new process was applied, which confirmed the usefulness and effectiveness of the treatment relative to the traditional process. Thus, the product obtained by the modified production process did not support the growth of L. monocytogenes.

Potential growth of Listeria monocytogenes in Italian mozzarella cheese as affected by microbiological and chemical-physical environment.

In the present study, 33 brands of mozzarella cheese (pasteurized cow milk mozzarella obtained by direct acidification through the addition of food-grade citric acid or obtained by natural acidification through the addition of thermophilic starter cultures, mozzarella for pizza mainly obtained by addition of citric acid, and pasteurized buffalo milk mozzarella obtained by adding microbial rennet) were characterized for the factors potentially influencing the growth of Listeria monocytogenes (microbial populations, moisture, pH, and organic acids). Then, the growth potential of L. monocytogenes in mozzarella was investigated by challenge tests performed at different temperatures. The presence of heterogeneous microflora (lactobacilli, streptococci, Pseudomonas spp., and, for buffalo mozzarella, yeasts) was evidenced. Almost all the product typologies were classified as high-moisture mozzarella cheese because moisture was >52%. Moreover, pH varied from 5.32 to 6.43 depending on the manufacturing methodology applied. Organic acid concentrations too showed great variability depending on the mozzarella production method, with values ranging from less than limit of detection (LOD; 16 mg/kg) to 14,709 mg/kg, less than LOD (216 mg/kg) to 29,195 mg/kg, and less than LOD (47 mg/kg) to 1,725 mg/kg in the water phase of lactic, citric, and acetic acids, respectively. Despite this presence, the concentration of undissociated acids was lower compared with the minimum inhibitory concentrations estimated for L. monocytogenes by other authors. This was confirmed by the results of the challenge tests conducted inoculating the pathogen in mozzarella produced with the addition of citric acid, as the microorganism grew fast at each temperature considered (4, 9, 15, and 20°C). Good hygiene practices should be strictly applied, especially with the aim of avoiding postproduction contamination of mozzarella, as the presence of organic acids and microflora is insufficient to prevent L. monocytogenes growth.

Effect of dairy product environment on the growth of Bacillus cereus.

pH is one of the most important parameters to manage bacterial replication in foodstuffs. In this study, the ability of 2 Bacillus cereus strains, 1 clinical human isolate (GPe2) and 1 isolate from a dairy product (D43), were investigated for in vitro growth at different pH values (from 3.5 to 7.5) at 2 temperatures (15 and 37°C), showing their ability to grow from 5.5 to 7.5 and from 5.0 to 7.5, respectively. The ability of spores of these 2 microorganisms to germinate in different typologies of dairy products (unflavored yogurt, Taleggio cheese, mascarpone cheese, and raw and pasteurized milk) was also investigated by inoculating the spores and maintaining the products at 15°C. No growth was observed in yogurt, likely due to the combined effect of low pH (<5) and the presence of natural microflora. An inhibitory action of the natural microflora on the growth of B. cereus was also hypothesized for Taleggio cheese and raw milk, as these substrates were characterized by a high natural lactic acid bacteria population and permissive pH values (5.8/6.8 in Taleggio cheese, >7 in raw milk). In pasteurized milk and mascarpone cheese, where pH was not restrictive for B. cereus growth and where no significant natural microflora was present, growth occurred rapidly up to loads close to 7 log cfu/g.

Former food products safety: microbiological quality and computer vision evaluation of packaging remnants contamination.

The use of alternative feed ingredients in farm animal's diets can be an interesting choice from several standpoints, including safety. In this respect, this study investigated the safety features of selected former food products (FFPs) intended for animal nutrition produced in the framework of the IZS PLV 06/14 RC project by an FFP processing plant. Six FFP samples, both mash and pelleted, were analysed for the enumeration of total viable count (TVC) (ISO 4833), Enterobacteriaceae (ISO 21528-1), Escherichia coli (ISO 16649-1), coagulase-positive Staphylococci (CPS) (ISO 6888), presumptive Bacillus cereus and its spores (ISO 7932), sulphite-reducing Clostridia (ISO 7937), yeasts and moulds (ISO 21527-1), and the presence in 25 g of Salmonella spp. (ISO 6579). On the same samples, the presence of undesired ingredients, which can be identified as remnants of packaging materials, was evaluated by two different methods: stereomicroscopy according to published methods; and stereomicroscopy coupled with a computer vision system (IRIS Visual Analyzer VA400). All FFPs analysed were safe from a microbiological point of view. TVC was limited and Salmonella was always absent. When remnants of packaging materials were considered, the contamination level was below 0.08% (w/w). Of note, packaging remnants were found mainly from the 1-mm sieve mesh fractions. Finally, the innovative computer vision system demonstrated the possibility of rapid detection for the presence of packaging remnants in FFPs when combined with a stereomicroscope. In conclusion, the FFPs analysed in the present study can be considered safe, even though some improvements in FFP processing in the feeding plant can be useful in further reducing their microbial loads and impurity.

Effects of antioxidant mixtures in the diet of finishing pigs on the oxidative status and shelf life of longissimus dorsi muscle packaged under modified atmosphere.

The effect of pig dietary supplementation with an antioxidant mixture (AOX), containing vitamin E and verbascoside, on animal oxidative status, meat quality parameters, and shelf life of the longissimus dorsi (LD) muscle was examined. Seventy pigs with an average live weight of 95.2 ± 1.2 kg were selected and assigned to 2 dietary treatments. The control (CTR) group was fed a commercial diet, and the AOX group was fed the same diet supplemented with the AOX, containing vitamin E and verbascoside from Verbenaceae extract, for 45 d before slaughter. At the beginning and at the end of the trial, blood samples were collected to determine oxidative status, using the Kit Radicaux Libres test. At slaughter, carcass weight was recorded and LD muscles from 10 pigs per treatment were sampled. Physical, chemical, microbiological, and sensory parameters and oxidative stability of LD muscle were assessed for up to 21 d of storage at 4°C under modified atmosphere packaging. Dietary AOX positively affected ( < 0.05) oxidative status and carcass dressing percentage. The oxidative and color stability of the LD muscle were improved ( < 0.05) in the AOX group compared with the control. The sensory shelf life revealed that at 15 d of storage, meat from the AOX group was comparable ( < 0.05) to the fresh meat in appearance and aroma. A lower ( < 0.05) spp. load was observed in the AOX samples than in the control samples. No other microbiological parameters were affected by dietary treatment. Overall, the present data showed that dietary AOX supplementation in pigs improved in vivo antioxidant status and exerted antioxidant and antimicrobial effects, thus enhancing the shelf life of raw pork under commercial conditions.

Microbiological shelf life at different temperatures and fate of Listeria monocytogenes and Escherichia coli inoculated in unflavored and strawberry yogurts.

Three different trials were performed on unflavored and strawberry yogurts produced in a small-scale dairy plant. In the first trial, the microbiological shelf life of the products was evaluated at 4, 8, and 20°C. At 4°C the product showed low total viable counts until the end of the trial (d 35=3.0±0.7 and 1.5±0.0 log cfu/g in unflavored and strawberry yogurt, respectively). The loads were lower in strawberry yogurt at 4°C compared with unflavored yogurt because of the antimicrobial activity exerted by potassium sorbate present in the fruit puree added. Yeasts were confirmed to be the specific spoilage agents of this product, reaching rapidly high loads with thermal abuse (5.9-7.4 log cfu/g at d 18). In the second trial, Escherichia coli and especially Listeria monocytogenes added at 2 concentrations (2 and 5 log cfu/g) showed a rapid decrease in both types thanks to the acidic conditions provided by the products, but L. monocytogenes was very resistant; its presence was always detected until the end of the period considered (d 68). In the third trial, no statistically significant differences were detected between wild and acid-adapted strains of L. monocytogenes added to the products, due to the quick adaptation that probably occurred after inoculation.