Italian Journal of Food Safety <p>The <strong>Italian Journal of Food Safety (IJFS)</strong> is the official journal of the <a href="" target="_blank" rel="noopener">Italian Association of Veterinary Food Hygienists (AIVI)</a>. The Journal addresses veterinary food hygienists, specialists in the food industry and other experts offering technical support and advice on food of animal origin. The <strong>Italian Journal of Food Safety</strong> publishes original research papers concerning food safety and hygiene, animal health, zoonoses and food safety, food safety economics. Reviews, editorials, technical reports, brief notes, conference proceedings, letters to the Editor, and book reviews are also welcome. Every article published in the Journal will be peer-reviewed by experts in the field and selected by members of the Editorial Board.</p> PAGEPress Scientific Publications, Pavia, Italy en-US Italian Journal of Food Safety 2239-7132 <p><strong>PAGEPress</strong> has chosen to apply the&nbsp;<a href="" target="_blank" rel="noopener"><strong>Creative Commons Attribution NonCommercial 4.0 International License</strong></a>&nbsp;(CC BY-NC 4.0) to all manuscripts to be published.<br><br> An Open Access Publication is one that meets the following two conditions:</p> <ol> <li>the author(s) and copyright holder(s) grant(s) to all users a free, irrevocable, worldwide, perpetual right of access to, and a license to copy, use, distribute, transmit and display the work publicly and to make and distribute derivative works, in any digital medium for any responsible purpose, subject to proper attribution of authorship, as well as the right to make small numbers of printed copies for their personal use.</li> <li>a complete version of the work and all supplemental materials, including a copy of the permission as stated above, in a suitable standard electronic format is deposited immediately upon initial publication in at least one online repository that is supported by an academic institution, scholarly society, government agency, or other well-established organization that seeks to enable open access, unrestricted distribution, interoperability, and long-term archiving.</li> </ol> <p>Authors who publish with this journal agree to the following terms:</p> <ol> <li>Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.</li> <li>Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.</li> <li>Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.</li> </ol> Whole genome sequencing based typing and characterisation of Shiga-toxin producing Escherichia coli strains belonging to O157 and O26 serotypes and isolated in dairy farms <p>In the present study, the genetic relationships as well as the virulome and resistome of newly sequenced O26 and O157 Shiga-toxin producing <em>E. coli</em> (STEC) isolates, collected from dairy farms in Italy, were investigated in comparison to publicly available genomes collected worldwide. The whole genome of Italian isolates was sequenced on Illumina MiSeq Platform. Reads quality control, <em>de novo</em> draft genome assembly, species confirmation and the 7-loci Multi-Locus Sequence Type assignment were performed using INNUca pipeline. Reference-based SNPs calling was performed on O157 and O26 genomes, separately, mapping contigs to high-quality finished genomes. Virulence and antimicrobial resistance determinants were detected <em>in silico</em><br>using the tool ABRicate. Phylogenetic reconstructions revealed that genomes clustered mainly based on their 7-loci MLST type. The virulome of tested genomes included 190 determinants. O157 genomes carried <em>chu</em> genes associated to heme mediated iron uptake, whereas O26 genomes harboured genes <em>ybt</em> associated to siderophore mediated iron uptake. Resistome analysis showed the presence of <em>tet</em>(34) on all but one O157 genomes and on only one O26 genomes. Only 4 genomes carried genes associated to multiresistance. In the present study, the genes <em>chu</em> and <em>ybt</em> were identified as potential biomarker for the differentiation of O157 and O26 serotypes.</p> Frederique Pasquali Federica Palma Marcello Trevisani Antonio Parisi Alex Lucchi Alessandra De Cesare Gerardo Manfreda ##submission.copyrightStatement## 2019-02-08 2019-02-08 7 4 10.4081/ijfs.2018.7673 Arcobacter spp. in bovine milk: An emerging pathogen with potential zoonotic risk <p>The aim of the present study was to assess the prevalence and genetic characteristics of <em>Arcobacter</em> spp. in bovine bulk tank milk produced in Apulia Region (Italy). Samples collected from 396 dairy farms, after enrichment in a selective broth, were subjected to an <em>Arcobacter</em> genus - specific Real Time PCR. Positive broths, previously filtered, were seeded on Karmali, MCCD and Columbia Blood Agar plates; presumptive <em>Arcobacter</em> spp. colonies were identified using an amplification and sequencing method and then characterized by Multi-Locus Sequence Typing (MLST). Prevalence of <em>Arcobacter</em> spp. in bovine milk samples was 5% (20/396); <em>A. butzleri</em> was the only isolated species, in agreement with previous studies that reported <em>A. butzleri</em> as the most commonly recovered species in milk and dairy products. MLST analysis of the 20 <em>A. butzleri</em> strains identified 81 alleles and 16 STs. Consistent with previous studies, MLST revealed a high level of heterogeneity between the <em>A. butzleri</em> isolates and confirmed the high discriminatory power of this method and its suitability for epidemiological investigations. This study confirmed the importance of raw milk as a possible source of <em>Arcobacter</em> spp. for humans.</p> Marta Caruso Laura Latorre Gianfranco Santagada Rosa Fraccalvieri Laura Maria Difato Angela Miccolupo Loredana Capozzi Elisabetta Bonerba Anna Mottola Antonio Parisi ##submission.copyrightStatement## 2019-02-08 2019-02-08 7 4 10.4081/ijfs.2018.7685 Detection of Extended- Spectrum Beta-Lactamase producing Escherichia coli from mesenteric lymph nodes of wild boars (Sus scrofa) <p>Wild boars (Sus scrofa) are increasing in several European countries, including Italy. In areas with intensive animal farming, like the Italian Emilia-Romagna region, they are likely to be exposed to antimicrobialresistant (AMR) bacteria of livestock origin. In 2017-2018, 108 mesenteric lymph nodes samples were collected from 108 wild boars hunted in Parma province, Emilia-Romagna region, to be tested for ESBL- and carbapenemase-producing <em>Escherichia coli.</em> One isolate (WB-21L) out of 108 (0.9%) was phenotypically confirmed as ESBLproducing <em>E. coli</em>. The strain WB-21L was tested by PCR for the genes <em>bla</em><sub>SHV</sub>, <em>bla</em><sub>CTX-M</sub>, <em>bla</em><sub>TEM</sub>, <em>bla</em><sub>AmpC</sub>, <em>bla</em><sub>KPC</sub>, <em>bla</em><sub>NDM</sub>, <em>bla</em><sub>VIM</sub>, <em>bla</em><sub>IMP</sub>, <em>bla</em><sub>OXA-48</sub>, <em>bla</em><sub>SPM</sub>, <em>bla</em><sub>BIC</sub>, <em>bla</em><sub>SIM</sub>, <em>bla</em><sub>DIM</sub>, <em>bla</em><sub>GIM</sub>, <em>bla</em><sub>AIM</sub>, resulting positive for TEM β-lactamase. Resistance to ampicillin, amoxicillin/clavulanic acid, streptomycin, sulfasomidine, tetracycline and trimethoprim confirmed the multi-resistance nature of the strain WB-21L. Nine <em>E. coli</em> isolates showed resistance to meropenem by the Kirby Bauer test but none of them showed Meropenem MIC values indicative of resistance. In conclusion, the present study shows the presence of ESBL <em>E. coli</em> in wild boars and the possible risk of transfer to game meat&nbsp;handlers and consumers. Future studies are needed to better evaluate the sources of AMR bacteria in wildlife.</p> Silvia Bonardi Clotilde Silvia Cabassi Simona Longhi Federico Pia Margherita Corradi Stefano Gilioli Erika Scaltriti ##submission.copyrightStatement## 2019-02-08 2019-02-08 7 4 10.4081/ijfs.2018.7707 Molecular identification of Sarcocystis spp. in cattle: partial sequencing of Cytochrome C Oxidase subunit 1 (COI) <p><em>Sarcocystis</em> spp. are protozoan parasites with an obligatory two-host life cycle, with herbivores as intermediate hosts and carnivores as definitive hosts. Cattle are intermediate hosts for several species of <em>Sarcocystis</em>: indeed, in addition to <em>S. cruzi, S. hirsuta</em> and <em>S. hominis</em>, at least four new species were recently identified in bovine muscle: <em>S. bovifelis, S. rommeli, S. bovini</em> and <em>S. heydorni</em>. Since is not possible to unambiguously discriminate between <em>S. hominis</em> and the new species either morphologically or by the analysis of the 18S ribosomial (rRNA) gene, the aim of the present study was to use molecular techniques to discriminate cattle <em>Sarcocystis</em> species, taking advantage of the higher discriminative power of the Cytochrome C Oxidase subunit I mitochondrial (mtDNA COI) gene. Therefore, 119 bovine muscle samples were tested to identify <em>S. hominis</em>-like sarcocystis using a multiplex PCR of the 18S rRNA gene; later, positive samples were tested using a newly designed primer set for the PCR amplification of COI gene. Species identification was achieved by sequencing the amplified products: 16 sequences were confirmed to belong to <em>S. bovifelis</em>, while 12 sequences didn’t constitute the best BLAST match of any of the published sequences, allowing to speculate the possible presence of <em>S. hominis</em>. This study confirms the higher discriminatory power of COI mitochondrial gene; besides, our work provides the first report of <em>S. bovifelis</em> in Italy.</p> Selene Rubiola Francesco Chiesa Stefania Zanet Tiziana Civera ##submission.copyrightStatement## 2019-02-08 2019-02-08 7 4 10.4081/ijfs.2018.7725 Experimental accelerated shelf life determination of a ready-to-eat processed food <p>The most direct way to estimate the shelf life of a product is to conduct simulation tests which are time consuming and expensive. Conversely, accelerated shelf life tests can be successfully used for stable products having long expected shelf life. The aim of the study was directed to verify the possibility to apply an accelerated shelf life test to perishable food products having a short-expected shelf life, such as a new ready-to-eat processed food preparation, composed mainly by cereals, tuna and chicken, packed in thermo-sealed trays and pasteurised. Different samples of the product were stored in thermal abuse conditions, collected periodically and subjected to determinations of TVB-N, pH and sensorial characteristics. Q<sub>10</sub> and activation energy were calculated allowing to obtain a predictive evaluation of the product shelf life at&nbsp;the 4°C recommended temperature. The product shelf life was assessed at 26 days <em>vs&nbsp;</em>the 30 days expected by the manufacturer, showing the possibility to apply successfully ASLT for products having short shelf life, saving both time and money.</p> M. Naceur Haouet Mauro Tommasino Maria Lucia Mercuri Ferdinando Benedetti Sara Di Bella Marisa Framboas Stefania Pelli M. Serena Altissimi ##submission.copyrightStatement## 2019-02-11 2019-02-11 7 4 10.4081/ijfs.2018.6919 Study on microbial communities in domestic kitchen sponges: Evidence of Cronobacter sakazakii and Extended Spectrum Beta Lactamase (ESBL) producing bacteria <p>Domestic environment, in particular, kitchen setting is a well-established source of microbial contamination. Kitchen sponges represent an important vehicle of microbial transmission and maintenance of spoilage bacteria and pathogenic strains responsible for food borne diseases. The aim of this study was to evaluate the microbial communities of 100 ‘in-use’ kitchen sponges, improving the knowledge on their role in cross-contamination in domestic environment and transmission of ESBLproducing strains. Sponges were processed for: aerobic mesophilic bacteria (AMB), <em>Enterobacteriaceae</em> (EB), yeasts and molds (YM), coagulase-positive staphylococci (CPS), micrococci (MCC), anaerobic sulfite reducing bacteria (ASR), and for the detection of <em>Listeria monocytogenes, Salmonella</em> spp. and <em>Yersinia enterocolitica</em>. A total of 309 enterobacteria strains were identified and then processed for ESBL (Extended Spectrum Beta Lactamase) phenotypical expression. A high contamination level of kitchen sponges was observed (mean value AMB 8.25±1.1; EB 5.89±1.2; YM 5.57±1.1; MCC 4.82±0.1 log CFU/g). Identified enterobacteria strains revealed several opportunistic and pathogenic agents such as <em>Enterobacter cloacae</em> (28%), <em>Citrobacter freundii</em> (23.3%), <em>Cronobacter sakazakii</em> (14.6%) and other strains in lower percentage. <em>Listeria monocytogenes</em> was found in only one sponge (1%). A total of 69 (22.3%) enterobacteria resulted ESBL+, with the following prevalence: P. rettgeri (50%), <em>L. adenocarboxilata</em> (30%), <em>K. pneumoniae</em> (25%), <em>K. oxytoca</em> (25%), <em>C. sakazakii</em> (20%), <em>E. cloacae</em> (20.7%), <em>C. freundii</em> (20.1%). Results confirm the potential role of kitchen sponges as vehicle for food-borne pathogens such as, <em>C. sakazakii</em> for the first time, infectious agents and spoilage microorganisms. The observed high contamination level and the presence of several ESBLs opportunistic pathogens, stresses the necessity to improve a proper education of the consumers on the effective treatment to reduce their microbial loads.</p> Stefania Maria Marotta Filippo Giarratana Anastasia Calvagna Graziella Ziino Alessandro Giuffrida Antonio Panebianco ##submission.copyrightStatement## 2019-02-11 2019-02-11 7 4 10.4081/ijfs.2018.7672 Microbiota analysis and microbiological hazard assessment in poultry carcasses from conventional and antibiotic free farms <p>The aim of this study was to assess microbiota and microbiological hazards in poultry carcasses from animals reared in conventional (n=15) and antibiotic free (n=15) farms. An aliquot of neck and breast skin was obtained from each individual carcass at the end of the refrigeration tunnel and submitted to DNA extraction. Total DNA was sequenced in the 16S rRNA and reads analysed with MG-RAST to classify the colonising bacteria up to the genus level and compare each taxonomic group in terms of mean relative frequency of abundance in conventional and antibiotic free carcasses. Firmicutes displayed abundances always higher than 38% but did not show statistically significative differences between conventional and antibiotic free carcasses. On the contrary, Bacteroidetes and Actinobacteria were significantly higher in antibiotic free then conventional carcasses (21.57 <em>vs</em> 10.95%; 19.29 <em>vs</em> 12.05%), whereas Proteobacteria were higher in the latter (33.19 <em>vs</em> 19.52%). The genera significantly higher in antibiotic free than conventional carcasses were <em>Chryseobacterium</em> (10.07 <em>vs</em> 1.94%), <em>Rothia</em> (3.08 <em>vs</em> 0.77%) and <em>Micrococcus</em> (1.12 <em>vs</em> 0.16%), while <em>Shewanella</em> was significantly higher in conventional carcasses (1.38 <em>vs</em> 0.26%). Among Firmicutes, the genera significantly higher in conventional carcasses were <em>Ureibacillus</em> (1.45 <em>vs</em> 0.11%) and <em>Bacillus</em> (3.28 <em>vs</em> 0.56%). The higher abundance of Proteobacteria in conventional carcasses might suggest that hygienic conditions in conventional farms are worse than antibiotic free farms. However, from a food safety point of view, <em>Salmonella</em> was not detected in both kinds of carcasses and the Campylobacter mean relative frequency of abundance was always lower than 0.4%.</p> Alessandra De Cesare Antonio Parisi Alex Lucchi Loredana Capozzi Angela Bianco Frederique Pasquali Gerardo Manfreda ##submission.copyrightStatement## 2019-02-11 2019-02-11 7 4 10.4081/ijfs.2018.7706 Detection of fish allergen by droplet digital PCR <p>Fish is one of fourteen allergens that must be highlighted on the label within the ingredients list. It should be noted that the European regulation, is very restrictive to allergens with zero tolerance. Therefore it is important to establish sensitive and specific methods for detecting fish allergen. Applicability to detect and quantify fish allergen by droplet digital polymerase chain reaction (ddPCR) has been evaluated in this work. Genomic DNA of three fish species belonging to the most common fish families were analyzed. PCR primers were designed to amplify a 166 bp region of the 18S rRNA gene. Comparative studies were performed to establish the optimal primer and probe concentrations.&nbsp; Annealing temperature was determined by using thermal gradient. The results have shown good applicability of the optimized 18S rRNA gene-method to detect and quantify small amounts of the target in all samples analyzed. However, validation studies are needed in order to apply ddPCR technology for routine allergens analysis.</p> <p><strong>&nbsp;</strong></p> Cinzia Daga Simona Cau Maria Giovanna Tilocca Barbara Soro Aldo Marongiu Bruna Vodret ##submission.copyrightStatement## 2019-02-11 2019-02-11 7 4 10.4081/ijfs.2018.7264 Food safety objectives, criteria, ranking and hierarchization <p>The paper describes the terminology of risk in the view of food safety objectives, criteria, ranking and hierarchization: the terms Performance Criterion, Process Criteria, Product Criterion, Microbiological Criteria, Food Safety Objective, Performance standard, Food safety policy objectives, Risk Assessment Policies, Weighing, Precaution, Prevention, Precautionary principle, Prevention principle, Principle of separation between risk assessment and management, Rank, Ranking, Categorization, Ranking, Priorization, ALARA (As Low Reasonably Achievable), ALOP (Appropriate level of protection), Risk Management, Risk management in the public function are reported and discussed.</p> Gaetano Liuzzo Stefano Bentley Federica Giacometti Silvia Piva Andrea Serraino ##submission.copyrightStatement## 2019-02-11 2019-02-11 7 4 10.4081/ijfs.2018.7395 Re: Effect of Nigella sativa against cisplatin induced nephrotoxicity in rats <p>n.a.</p> Mehdi Nematbakhsh ##submission.copyrightStatement## 2019-02-08 2019-02-08 7 4 10.4081/ijfs.2018.7780