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In the period 2006-2011 six public catering establishments (3 canteens and 3 cafeterias) were monitored, trough audit and sampling, in order to verify the application of good manufacturing and hygiene practices during food production, handling and serving. The compliance to microbiological food safety criteria (Listeria monocytogenes and Salmonella spp.) and process hygiene criteria were investigated using ISO standards for microbiological analyses. A total of 612 samples were collected: 192 food samples and 288 environmental swab samples from canteens; 33 food samples and 99 swab samples from cafeterias. Regarding food safety, two samples were in disagreement with criteria fixed in EU Regulation as Listeria monocytogenes was isolated from a turkey breast in a canteen and from a sandwich in a cafeteria. Regarding process hygiene criteria, as no microbiological limits are legally defined for catering services, for this study limits were fixed according to the quality standards of tender, scientific literature and laboratory experience. 23.4% foodstuffs and 8.7% swabs resulted non-compliant in canteens; 48.5% foodstuffs and 6.1% swabs resulted non-compliant in cafeterias. The count of coagulase-positive staphylococci (CPS) was higher of the fixed limits in raw turkey meat and in cooked spinach: the presence among CPS of S. aureus was confirmed, strains were not able to produce enterotoxins. The most common non-compliance in hygiene criteria was represented by aerobic colony count (60.7% of total non-compliance in canteens and 75.0% in cafeterias) and coliform bacteria (20.3% in canteens and 25.0% in cafeterias). Nine raw foods or raw readyto- eat food samples were non-compliant for both coliform bacteria and aerobic count; one sample (raw turkey meat) was non-compliant for CPS and aerobic count but resulted to be compliant after cooking. Auditing and sampling are the most effective tools to improve food quality standard and to enhance food business operators’ awareness of hygiene procedures.

Salmonella bongori 48:z35:– is considered endemic to Sicily (Italy) due to its epidemiological peculiarity. To our knowledge, no previous cases of human infection caused by S. bongori 48:z35:– have ever been reported in mainland Italy. Here we describe the isolation of S. bongori 48:z35:– from a 1-year-old symptomatic child in northwest Italy (Piedmont Region). The strain showed no antimicrobial resistance. Reporting of S. bongori 48:z35:– in a previously safe area is important to identify epidemiological changes.

Aflatoxins (AFs) are mycotoxins produced by some species of Aspergillus. In dairy cows, ingested AFB1 is metabolized into carcinogenic AFM1 which is eliminated through milk, thus posing a risk for consumer health. Here we describe the set, validation, and application of screening (ELISA) and confirmatory (HPLC) tests carried out on milk samples collected through official control of mycotoxin levels in northern Italy over a three-year period (2012–2014). The limit of detection (LOD) was set at 5 ppt (ng/kg) and 2 ppt for ELISA and HPLC, respectively, and the limit of quantification (LOQ) was 10 ppt for confirmatory HPLC. A total of 1668 milk samples were analyzed: ELISA identified 36 (2.2%) positive milk samples that were subsequently confirmed by HPLC. The level of AFM1 in the positive samples ranged between 18 ± 2 and 208 ± 27 ppt. Of the total samples, only eight (0.5%) were found non-compliant with the EU regulatory limit (50 ppt; range 74 ± 10 to 208 ± 27 ppt). Use of ELISA and HPLC tests in series allows for high-volume analysis of samples, thus saving time and money while guaranteeing high analytical precision and accuracy.

The aim of the study involved evaluation of the presence of aflatoxin M1 in milk for sale in a specific North West Italian region, Piedmont. The study, conducted from November 2003 to July 2005, was linked to the specific emergency situation which arose due to the climatic conditions during the summer of 2003 which encouraged the development of aflatoxin B1 in items used for animal feed. This in turn led to the transfer of the metabolite, aflatoxin M1, into the milk. In total some 316 milk samples were collected during the commercial phase by the official control bodies and analysed. The analysis involved the use of high pressure liquid chromatography (HPLC) combined with fluorimetric measurement, and purification of the extracts using immunoaffinity columns. The results indicated only 2 non conforming samples (0.6%), with limits higher than those set out in the regulations (0.05 μg L−1). In addition, the analyses revealed, in 5 samples (1.6%), threshold values of 0.05 μg L−1. From the data obtained it can be seen that the “aflatoxins” problem only marginally affected Piedmont Region though the trend for average monthly values suggests a return to the use of contaminated animal feed as soon as official controls are less intensive.

In the latest year, and also in 2013, Salmonella was the most frequently detected causative agent in foodborne outbreaks (FBOs) reported in Europe. As indicated in EFSA report (2015) the serotypes mostly associated to FBOs are S. Typhimurium and Enteritidis; while Salmonella Typhimurium is generally associated with the consumption of contaminated pork and beef, FBOs due to Salmonella Enteritidis are linked to eggs and poultry meat. In this study it is described the investigation of a domestic FBO involving four adults and linked to homemade lasagne. Investigations were performed to determine the relatedness of Salmonella strains, identify the sources of infection, and trace the routes of Salmonella contamination in this FBO. Salmonella strains were isolated in 3 out of 4 patient stool samples and from lasagne and all of them were serotyped as S. Enteritidis. Pulsed-field gel electrophoresis (PFGE) analysis revealed the genotypical similarity of the all the strains. Although serotyping and PFGE analysis identified the common food source of infection in this FBO, it was not possible to determine how or at what point during food preparation the lasagne became contaminated with Salmonella.

Cardiac syncope heralds significantly higher mortality compared with syncope due to non-cardiac causes or unknown aetiology, which is commonly considered a benign event. Nonetheless, previous studies have typically investigated the outcome of syncope across different aetiological subgroups, rather

Staphylococcal food poisoning is a common foodborne disease caused by the ingestion of staphylococcal enterotoxins (SEs) produced mainly by enterotoxigenic strains of Staphylococcus aureus. To date, 21 SEs and/or enterotoxin-like types have been identified, several of which represent a potential hazard for consumers. To protect consumer health and to reduce the amount of SE-contaminated food entering the market, European Union legislation regulating food safety requires testing for SEs. The Italian National Reference Laboratory organized a ring trial to test technical and analytical proficiency in the national network of official food laboratories. Twenty-four laboratories took part, and each received and analyzed 24 blind dairy samples. Reproducibility of the results from the laboratories was assessed by the Cohen k index, and accuracy (sensitivity and specificity) was evaluated according to the International Organization for Standardization definition (ISO 16140:2003). Trial results revealed partially satisfactory agreement: 254 of 276 possible paired participants (92%) reached a k value0.60, which is conventionally recognized as satisfactory. Accuracy was deemed satisfactory; 100% sensitivity and 100% specificity were achieved by 22 and 18 of the 24 laboratories, respectively.

Several EC Directives have been promulgated to protect allergic individuals but no rule has been established with regard to allergen cross-contamination caused by shared transport vehicles or common processing equipment. The aim of this research was to quantify, by enzyme-linked immunosorbent assay (ELISA) or real-time polymerase chain reaction, the presence in meat- or fish-based foods of four allergens (milk, egg, crustaceans and molluscs) that was not indicated either in the list of ingredients or in the label alert. In the time frame of 2007–2009, a total of 723 samples were subjected to 1983 analyses. The percentage of samples scoring positive ranged between 1.8% and 6.8% over the 3 years, and the concentrations of undeclared allergens found were 0.3–13.3 mg kg−1 for milk (β-lactoglobulin) and 0.21–12 mg kg−1 for egg white proteins. On this basis, the possibility of cross-contamination serious enough to raise public health concern cannot be dismissed.

Following the detection of methicillin-resistant Staphylococcus aureus (MRSA) ST398 in food-producing animals, both livestock and wildlife, and derived products, are considered potential sources of MRSA in humans. There is a paucity of data on MRSA in foods in Italy, and the data regarding wild animals are particularly scarce. A total of 2162 food samples collected during official monitoring activities in 2008 were analyzed for the detection of S. aureus. Also, samples from 1365 wild animals collected by the National Reference Center for Wild Animal Diseases in 2003-2009 were subjected to anatomopathological examination. S. aureus isolates were processed for phenotypic and molecular methicillin resistance determinations. S. aureus was found in 2.0% of wild animal carcasses and in 3.2% of wild boar lymph nodes: none showed methicillin resistance. The prevalence of S. aureus in food was 17.1%. Two MRSA strains, both from bulk tank milk (prevalence 0.77%) were isolated: the strains were resistant to tetracycline, had spa-type t899, and were negative for the Panton-Valentine leukocidin gene. The low prevalence of MRSA suggests that the risk of transmission to humans via food is limited. However, attention should be paid to the cattle food chain, which may be a potential route of transmission of LA-MRSA.

Campylobacteriosis was the most frequently reported zoonosis in the European Union (EU) in 2010, with Campylobacter jejuni, Campylobacter coli, and Campylobacter lari as the most frequently reported species in foodborne outbreaks (FBOs). Relatively sensitive to environmental factors, these species may be present in low numbers. In line with EU policy for food control and FBO detection and in view of the need to reduce response time, we validated an alternative molecular method according to ISO 16140:2003 which establishes the general principle and technical protocol for the validation of alternative methods in the microbiological analysis of food. We used a qualitative real-time PCR commercial kit for the detection of C. jejuni, C. coli, and C. lari in two food categories "fruit and vegetable-based products" and "dairy products". The validation protocol comprises two phases: the first is a method comparison study of the alternative method against the reference method, and the second is an interlaboratory study of each of the two methods. In the first step, ISO 16140:2003 validation examines the following parameters: limit of detection (LOD); relative accuracy, relative specificity and sensitivity; relative detection level (RDL); and inclusivity and exclusivity. Except for LOD, inclusivity and exclusivity, the other steps were performed against the reference method (ISO 10272:2006). The LOD of the real-time PCR method was set at 4CFU/25g or mL for both food categories. Relative accuracy (98.33%), specificity (96.77%), and sensitivity (100%) were recorded for the food category "fruit and vegetable-based products" and 93.3%, 88.24%, 100%, respectively, for "dairy products". The RDL according to Fisher's exact test was p=1 for both food categories, for each level, and each food/strain combination. The interlaboratory study results showed correct identification of all 24 blind samples with both methods by all the participating laboratories. The results show that this commercial kit is suitable for the rapid detection of C. jejuni, C. coli, and C. lari on fruit, vegetables and dairy products and may aid in official controls. In conclusion, the use of alternative methods is recommended for the rapid identification of positive samples and the identification of the possible bacterial source in a FBO within 48 h.

UNLABELLED Staphylococcal foodborne intoxication, occurring after consumption of staphylococcal enterotoxins (SEs) in food, is considered one of the most common forms of bacterial foodborne outbreaks worldwide. Milk and dairy products account for 5% of all the incriminated foods in staphylococcal outbreaks, referring to Europe. The distribution of genes encoding for enterotoxins in Staphylococcus aureus strains is highly variable, with some carried on stable regions of the chromosome and others carried on mobile genetic elements. The aim of this study was to analyse the distribution of genes encoding for SEs in Staph. aureus strains isolated from milk and dairy products. In the period from January 2010 to June 2011, a total of 1245 dairy samples (848 of raw milk and 397 of dairy products) were collected and analysed for detection of genes encoding for 11 SEs and SEls (SEA, SEB, SEC, SED, SEE, SEG, SEH, SEI, SER SElJ and SElP) according to the procedures of the Italian National Reference Laboratory for coagulase-positive Staphylococci including Staph. aureus. Staphylococcus aureus strains were isolated in 481 (39%) samples. Of the 481 isolates of Staph. aureus tested, 255 (53%) were positive for one or more SE genes, and thirty-five different enterotoxin gene profiles were distinguished among the isolates. ser gene, found in 134 (28%) of the isolates, was the most frequent, followed by sed (25%) and selj genes (25%). The identification of new SEs increased the isolation frequency of enterotoxigenic staphylococci, thus suggesting that the pathogenic potential of Staph. aureus may be of greater importance than previously thought. Further studies are needed to quantify the expression of these new enterotoxins, and to assess their contribution to foodborne disease burden. SIGNIFICANCE AND IMPACT OF THE STUDY The analyses targeted 11 staphylococcal enterotoxins genes and 35 different enterotoxin gene profiles were distinguished among the isolates. A total of 255 Staph. aureus isolates were positive for one or more SE genes while ser gene was the most prevalent. In 93% of the isolates bearing genes located on the enterotoxin gene cluster (n = 89), both seg and sei genes were present.

Campylobacteriosis was the most commonly reported zoonosis for confirmed human cases in European Union during 2011. Poultry meat was very often implicated in Campylobacter infections in humans. In Italy commerce of raw poultry meat is common in open-air markets: these areas can be considered at high risk of bacterial contamination due to the high presence birds like pigeons. The aim of this study was to collect data about the contamination by thermotolerant Campylobacter of raw poultry meat commercialised in open-air markets, of work-surfaces in contact with poultry meat and of pigeon stools sampled in the market areas in Turin, Northern Italy. Between September 2011 and December 2012, 86 raw poultry meat samples, 86 environmental swabs and 108 animal samples were collected in 38 open-air markets. Analysis were carried out according to ISO10272-1:2006 standard. C.coli was detected in 2.3% (2/86) of raw poultry meat samples, whereas no swab (0/86) resulted positive. Of pigeon stool 28% (30/107) was positive for C.jejuni (83.3% C.jejuni subsp. jejuni and 16.7% C.jejuni subsp. doylei). C.jejuni subsp. jejuni was isolated from 1 dead pigeon. Our results showed lower rates of contamination than those reported at retail in Europe. Although samples were collected in areas at high risk of contamination, raw poultry meat and work surfaces reported a low level of presence of thermotolerant Campylobacter. The high percentage of C.jejuni isolated from pigeon stools showed the importance of a continuous application of preventive measures by the food business operators and the surveillance activity by the Competent Authority.

The effect of Lactococcus lactis nisin-producing strains, isolated from Italian fermented foods, onthe survival of two foodborne pathogens namely Listeria monocytogenes and Staphylococcus aureuswas investigated in experimental cheese production. One of the three Lactobacillus lactis nisin inn-oculated as starters, Lactobacillus lactis 41FL1 lowered S. aureus count by 1.73 log colony-formingunits (cfu)/g within the first 3 days, reaching the highest reduction, 3.54 log cfu/g, by the end ofripening period of 60 days. There was no effect on L. monocytogenes. The application of L. lactis41FL1 as bioprotective culture in controlling S. aureus shows considerable promise.Keywords Lactococcus lactis, nisin, Staphylococcus aureus, Listeria monocytogenes, Cheese.INTRODUCTIONLactic acid bacteria (LAB) are widely used forthe fermentation and preservation of a widerange of milk, meat and vegetable foods.Among LAB strains, Lactococcus lactis isextensively used as starter culture in the dairyindustry, and the potential applications of thesestrains in food production are well documented.In particular, in recent years, great interest wasfocused on the possible use of certain productsof their metabolism, called bacteriocins, aspotential biopreservatives in foods (O’Sullivanet al. 2002).Bacteriocins are proteinaceous antimicrobialcompounds ribosomially synthesised by bacteriaand able to inhibit a high number of potentialpathogens among which Listeria monocytogenesand Staphylococcus aureus (Cotter et al. 2005).The current technologies employed to inacti-vate bacterial pathogens in foods are not alwaysefficient and often represent particular treatmentsthat may modify the fisical and chemical proper-ties of the food products. Therefore, newapproaches to combat pathogenic and spoilagemicroorganisms are needed. In the last fewyears, bacteriocins from LAB have been thor-oughly characterised and tested in food systemsas new food preservatives (Stiles 1996).Although many bacteriocins have been charac-terised, two of the most widely used in foodindustry as biopreservative are nisin (Class I,lantibiotic) and pediocin (Class II). Nisin is thebacteriocin with the longest history of safe usein food industry. It has been shown to be effec-tive for the microbial control in a number ofdairy products and also widely assessed incheese manufacturing (Ross et al. 2002). Inmost of cases, the bacteriocin is incorporatedinto the product as a dried concentrated powder(O’Sullivan et al. 2002), but currently, the useof nisin-producing starter or costarter culturesseems to be an alternative way of incorporatingthe bacteriocin (e.g. cheesemaking process)(Sobrino-Lopez and Marton-Belloso 2008).Among the pathogenic bacteria potentiallyassociated with dairy foods, S. aureus andL. monocytogenes are the cause of numerous

Between June and September 2010, widespread Italian consumer reports of unusual blue spoilage on fresh dairy products were publicized, resulting in the so-called blue mozzarella event. An inordinately high number of samples from mozzarella and whey cheese products of Italian and German production subsequently tested positive for Pseudomonas fluorescens. The aim of this study was to verify whether a selected P. fluorescens strain was responsible for this apparently unusual event. Molecular characterization of 181 isolated P. fluorescens strains was conducted using a newly optimized pulsed-field gel electrophoresis protocol. Although a high number of pulsotypes was found (132), only four pulsotypes were associated with more than one production plant, and only one German isolate had the same pulsotype as was detected in two Italian plants. This is the only evidence of possible cross-contamination among cheeses from the two countries. The overall results did not support the spread of contamination from German to Italian plants or the presence of one environmental strain that spread in both countries.

The autochthonous “Piemontese” bovine breed is primarily raised for meat due to the high quality of production. The aim of this study was to investigate the microbiological, nutritional and chemical aspects of samples meat from cattle breeding “Piemontese”.

L. monocytogenes represents a primary concern in the production of Gorgonzola, a Protected Designation of Origin (PDO) Italian blue-veined cheese produced only in the Piedmont and Lombardy regions. L. monocytogenes isolates (N = 95) obtained from Gorgonzola rinds, paste, and production/ripening environments were serotyped and then genotyped using Pulsed Field Gel Electrophoresis (PFGE). The goal of this study was to investigate the variability of L. monocytogenes PFGE-types across different PDO Gorgonzola manufacturers (N = 22). The majority of the strains (88%) were serotyped as 1/2a. PFGE identified 2 major pulse-types grouping 62 strains, detected from different plants and years, suggesting the presence of persistent and niche-adapted L. monocytogenes. In 9 plants, environmental strains shared the same pulse-types with strains from rinds or paste, suggesting a possible transmission pathway. Encouragingly, L. monocytogenes was retrieved from only 1 paste, indicating that production processes were under control in 21 plants. In the remaining plant, un-effective pasteurization or cross-contamination during production processes could be the cause of the contamination. Consequently, it is imperative that producers operate under the total respect of the Good Manufacturing Practices and following the principles of the Hazard Analysis Critical Control Point plans, in order to contain contamination throughout the whole processing.

Twenty-five yeast strains isolated from sourdough samples for Panettone, Pandoro and Cornetto brioche manufactured by eight different bakeries in northern Italy were characterised. Classification was performed by the simplified identification method (SIM), Kurtzman and Fell's identification protocol, the API system from bioMerieux (France) and the MicroLog system from Biolog (USA). Genetic diversity was investigated by randomly amplified polymorphic DNA fingerprinting and mitochondrial-DNA restriction enzyme analysis. Sequences of the internal transcribed spacers between 18S and 26S rDNA genes were analysed. Candida humilis was the predominant species (56% of isolates), whereas the remaining strains (44%) were related to the Saccharomyces cerevisiae sensu stricto group. Identification systems based on phenotypic analysis proved to be unreliable to identify yeasts from sourdough. Either RAPD-PCR or mtDNA restriction analysis showed to be suitable for the identification of species, but could not be used to differentiate among the isolates at the strain level. Sequencing of the ITS region permitted a consistent classification of the sourdough yeasts.

Contaminants like heavy metals could remain in milk and its by-products if forage used for ruminant nutrition is contaminated with high levels of these elements. In EU maximum levels (LMR)are established only for lead in milk. In Russia limits have been fixed for different heavy metals in milk and dairy products; for this reason exporting countries, including Italy, have to demonstrate the conformity of their products to the expected limits. IZSPLV analysed 18 cheese samples to research lead, cadmium, arsenic and mercury. All samples were negative and could be exported toRussia.

Raw milk at vending machine is surging in popularity amongst consumers of Northern Italy; indeed in Piedmont Region there are more than 100 vending machines. In June 2008 Piedmont Region set out a specific monitoring plan to check the milk quality. From June to December 2008, 113 raw milk samples were collected at vending machines. Samples were analysed for Listeria monocytogenes, Salmonella spp., coagulase positive staphylococci, Staphylococcus aureus and Campylobacter. Moreover, 100 samples were analysed for the quantification of aflatoxin M1. 26 samples have been resulted Not Conform for the hygienic criteria and 1 exceeded the aflatoxin M1 limit.

Veterinary Record (2004)155, 337-338L Decastelli, DVM,G. Ru, DVM,S. Gallina, DVM,Istituto ZooprofilatticoSperimentale delPiemonte, Liguria e Valled'Aosta, Via Bologna 148,10154 Turin, ItalyG. Brizio , DVM,D. Gentile, DVM,Servizio di prevenzione,ASR 17, Via Bava S. Paolo40,12045 Possano, (GN),ItalyA . Caprioli, PhD,Istituto Superiore diSanita, Viale Regina Elena299,00161 Rome, Italy

The aim of this study was to compare two different methods for enterotoxic Staphylococcus aureus characterization. 110 S.aureus strains was isolated from foods and tested with ELISA method able to detect toxins type A to E in culture medium and PCR protocols able to detect the presence of genes (sea to see; seg to sej; sep; ser) encoding for staphylococcal enterotoxins. 27 strains came out positive with ELISA; 68 resulted to have at least one encoding gene. sea and ser genes were detected respectively in 29,1% and 27,3% of strains.