Evaluation and validation of an alternative method to detect Campylobacter spp. in dairy products

Foods implicated in human campylobacteriosis include raw or undercooked poultry and raw dairy products. Because Campylobacter spp. are the most frequently reported cause of bacterial infection in the European Union and because conventional methods are cumbersome, rapid methods for Campylobacter detection and quantification in food are needed. With this study we sought to validate, according to the standard procedure (UNI EN ISO 16140:2003), an alternative to the reference analytical method (UNI EN ISO 10272- 1:2006) for official controls of Campylobacter spp. in raw milk and dairy products. Milk samples collected from 16 milk vending machines located throughout the Genoa metropolitan area were analyzed using two different methods, an enzymelinked fluorescent assay (ELFA) and a real-time PCR assay, and evaluated in parallel against the reference method. In addition, a total of 460 samples of raw milk collected from milk vending machines were analyzed by ELFA. Results obtained with ELFA showed it was compliant with UNI EN ISO 10272-1:2006 criteria and that the immunoassay had 100% sensitivity, specificity, and accuracy. Regarding samples of milk vending machines, 5.0% (23/460) tested positive at ELFA screening and were subsequently confirmed as C. jejuni. Validation according to UNI EN ISO 16140:2003 of the ELFA method suggests it may be a useful alternative to conventional methods for detecting Campylobacter spp. in official controls

Development and validation of species-specific molecular diagnostic tool for Opisthorchis felineus (Digenea, Opisthorchiidae) metacercariae

Opisthorchis felineus (family Opisthorchiidae) is a parasitic flatworm representing a serious threat to humans in some countries. Opisthorchiasis occurs after consumption of raw or undercooked cyprinid fish infected by the metacercarial stage of the parasite. Due to its small size, detection of the parasite in fish fillet is time-consuming and difficult. Furthermore, isolated metacercariae can be identified to genus but not to species level using morphological features and molecular techniques are necessary. In this work, we describe the development of primers for a diagnostic PCR amplification of a 254-bp fragment of the cytochrome c oxidase I in the mitochondrion of Opisthorchis felineus metacercariae isolated from fish fillet, together with a validation protocol for this method

Enterotoxigenic strain of Staphylococcus aureus causing food-borne outbreak in a private context

In the last European Food Safety Authority (EFSA) report on zoonoses a total of 5262 food- borne outbreaks (FBOs) have been reported in Europe in 2010. Staphylococcal FBOs are caused by consuming food contaminated with one or more preformed enterotoxins and are characterised by rapid onset of symptoms. In May 2012, an Italian family made up of five people was involved in a FBO: food sample of arancini (fried rice balls) were analysed and resulted positive for coagulase positive staphylococci (CPS) (>100,000 cfu/g) and for staphylococcal enterotoxins (SE) (types A and C). Laboratory analyses also led to the isolation of Staphylococcus aureus strain carrying the gene encoding for enterotoxin type A and belonging to the human biotype. The FBO described in this paper should be included in the next official FBO report as a strong evidence case: food and toxins responsible for symptoms and enterotoxigenic S. aureus strain were identified and the clinical symptoms matched with the final diagnosis

A Salmonella enterica subsp. enterica serovar Enteritidis foodborne outbreak after consumption of homemade lasagne

In the latest year, and also in 2013, <em>Salmonella</em> 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 <em>S.</em> Typhimurium and Enteritidis; while <em>Salmonella</em> Typhimurium is generally associated with the consumption of contaminated pork and beef, FBOs due to <em>Salmonella</em> 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 <em>lasagne</em>. Investigations were performed to determine the relatedness of <em>Salmonella</em> strains, identify the sources of infection, and trace the routes of <em>Salmonella</em> contamination in this FBO. <em>Salmonella</em> strains were isolated in 3 out of 4 patient stool samples and from <em>lasagne</em> and all of them were serotyped as S. Enteritidis. Pulsed-field gel electrophoresis (PFGE) analysis revealed the genotypical similarity of 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 <em>lasagne</em> became contaminated with <em>Salmonella</em>

Evidence of Antimicrobial Resistance and Presence of Pathogenicity Genes in Yersinia enterocolitica Isolate from Wild Boars

Yersinia enterocolitica (Ye) is a very important zoonosis andwild boars play a pivotal role in its transmission. In the last decade, the wild boar population has undergone a strong increase that haspushed them towards urbanized areas, facilitating the human–wildlife interface and the spread of infectious diseases from wildlife to domestic animals and humans. Therefore, it is important to know the serotype, antimicrobial resistance and presence of pathogenicity genes of Yersinia enterocolitica (Ye) isolated in species. From 2013 to 2018, we analyzed the liver of 4890 wild boars hunted in Liguria region; we isolated and serotyped 126 Ye positive samples. A decisive role in the pathogenicity is given by the presence of virulence genes; in Ye isolated we found ystB (~70%), ymoA (45.2%), ail (43.6%) and ystA (~20%). Moreover, we evaluated the susceptibility at various antimicrobic agents (Ampicillin, Chloramphenicol, Enrofloxacin, Gentamicin, Kanamycin, Trimethoprim–Sulfamethoxazole, Sulfisoxazole, Ceftiofur and Tetracycline). The antibiotic resistance was analyzed, and we found a time-dependent increase. It is important to shed light on the role of the wild boars as a reserve of potentially dangerous diseases for humans, and also on the antibiotic resistance that represents a public health problem

Food allergens: state of the art in Piedmont region in the period 2011-2012

The US National Institutes of Allergy and Infectious Diseases defines food allergy as <em>adverse health effect arising from a specific immune response that occurs reproducibly on exposure to a given food</em>. Undeclared allergens in food label represent a risk for consumers, as there is no therapy for food allergies. According to Directive 2003/89/EC, declaration of all ingredients and derived substances in the label is mandatory. In 2011-2012, in Piedmont region (North-western Italy) 285 food samples were analysed for β-lactoglobulin and 234 for egg proteins. The aim of this work was to analyse 2 years data in order to assess the presence of undeclared milk and egg allergenic proteins in food placed on the market checking the compliance of labeling of food allergens. Analyses were carried out with ELISA tests, both for the detection of the egg and milk proteins. β-lactoglobulin was found in 2.8% (8/286) of samples, while egg proteins in 4.7% (11/234)