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Several food producers use precautionary allergen labelling (PAL) to alert consumers to the possible unintended presence of allergens in food products. In the vast majority of countries, the use of...

Fighting food frauds is a ceaseless challenge because of the constant evolution of fraudulent practices and for the consequences both on consumers’ and on globalized trade. In Italy fish is a vulnerable commodity for frauds thanks to the high national production, importation and consumption and it is important to monitor the entire food chain in order to detect and prevent fraudulent actions, such as species substitutions, which is considered the most common fraud in seafood. Aim of this study was to realise a targeted Monitoring Plan to estimate the prevalence of fish species substitutions in Italy. As a first step, Italian fish supply chain, from production to selling, was analysed, in order to identify products and chain points at risk, by reviewing literature and by involving, in two focus groups, food inspectors and representatives of the large scale food distribution system. Then a monitoring plan was designed by sampling three fish species considered at major risk for their economic value and/or large consumption, i.e. tuna, grouper and flat fishes, at different selling points (wholesale markets, retail markets, fish shops and supermarkets). From February to March 2017, 242 samples from fresh, frozen or transformed whole fishes or fillets were sampled in 13 Italian cities (5 in Northern Italy, 2 in Central Italy, 3 in Southern Italy and 3 in the main Islands). Samples were analysed by “FINS” (Forensically Informative Nucleotide Sequencing), using two markers: cytochrome oxidase subunit I gene (COI) as first option and then mitochondrial cytochrome B gene (cytb), if necessary to identify uncertain or unassigned samples. Species substitutions were uncovered in 8.7% of analysed samples, principally related to grouper (prevalence of 14.71%). Supermarkets resulted the selling points with a major number of frauds (prevalence of 12.79%). Substituted species were taxonomically related to those declared on the label and no species harmful for consumers were detected. Results obtained can give to National Authorities a detailed frame of trends in fish substitution frauds in Italy, providing also relevant information to put into effect control measures.

Rationale Carry-over is an undesirable contamination from medicated to non-medicated during the production of feedingstuffs. In 2014 the European Parliament and the Council started working to produce a new regulatory act that will fix tolerable levels of drugs by carry-over in non-target feed to have a harmonized practice to evaluate this contamination by veterinary drugs. Methods We developed a rapid and effective multi-analyte method coupling ultraperformance liquid chromatography to tandem mass spectrometry (UPLC/MS/MS) for the detection of 37 drugs belonging to different classes of antimicrobials (sulfonamides, tetracyclines, macrolides, quinolones, pleuromutilins and streptogramins) in feeds at carry-over levels. The method was in-house validated in the concentration range 0.25-2.0 mg kg-1 , according to the Regulation (UE) 2017/625 requirements and the guideline included in the Commission Decision 2002/657/EC for official methods. Results The UPLC/MS/MS method allows the determination of the antimicrobials in 15 min, by providing results compliant to the criteria established by the European Commission legislation. All the analytes showed a limit of detection (LOD) in the range 2.0-5.0 μg kg-1 and a limit of quantification (LOQ) at 10.0 μg kg-1 ; oxytetracycline, doxycycline, spiramycin and virginiamycin have a higher LOD and LOQ (15.0 μg kg-1 ; 30.0 μg kg-1 , respectively). Recoveries were satisfactory ranging from 90.4% to 103.1%. Conclusions The method is characterized by an effective clean-up of all drugs without the use of large sample size and organic solvent extraction.

Cd bioaccumulation was investigated in the most consumed rice varieties (Oryza sativa L.) in Italy to assess the health risk related to consumption of the cereal for Italian population. In addition, the health implications concerning to consumption of rice-based foods was evaluated for Italian infants. Cadmium was detected by Q-ICP-MS. Method was validated in compliance with UNI CEI EN ISO/IEC 17025. Cd concentrations were found below the Maximum Levels (MLs) established at the European Union level for rice (0.092 mg kg−1 for brown rice and 0.062 mg kg−1 for white rice). The Cd Estimated Weekly Intakes (EWIs) through brown rice and white rice consumption by total population accounted approximately for 6% and 4% of the TWI set by the European Food Safety Authority (2.5 μg kg−1 body weight), respectively. The Cd EWIs through rice consumption by different population groups obtained in this survey did not show notable differences with those related to the total population except for child group that showed the highest EWIs of Cd. As regards to the Cd EWIs quantified for infants of four, six and eight month old considering the rice-based infant food consumption accounted for 22%, 38%, and 36% of the EFSA TWI, respectively.

Background and aim Sarcoidosis is a systemic granulomatous inflammatory disease whose causes are still unknown and for which epidemiological data are often discordant. The aim of our study is to investigate prevalence and spatial distribution of cases, and identify environmental exposures associated with sarcoidosis in an Italian province. Methods After georeferentiation of cases, the area under study was subdivided with respect to Municipality and Health Districts and to the altitude in order to identify zonal differences in prevalence. The bioaccumulation levels of 12 metals in lichen tissues were analyzed, in order to determine sources of air pollution. Finally, the analysis of the correlation between metals and between pickup stations was performed. Results 223 patients were identified (58.3% female and 41.7% male of total) and the mean age was 50.6±15.4 years (53.5±15.5 years for the females and 46.5±14.4 for the males). The mean prevalence was 49 per 100.000 individuals. However, we observed very heterogeneous prevalence in the area under study. The correlations among metals revealed different deposition patterns in lowland area respect to hilly and mountain areas. Conclusions The study highlights a high prevalence of sarcoidosis cases, characterized by a very inhomogeneous and patchy distribution with phenomena of local aggregation. Moreover, the bioaccumulation analysis was an effective method to identify the mineral particles that mostly contribute to air pollution in the different areas, but it was not sufficient to establish a clear correlation between the onset of sarcoidosis and environmental risk factors.

Histamine content was tested on 311 samples of fresh fish and fish products collected in Puglia (Italy) in the years 2009–2011. The analyses were carried out employing ELISA for screening and HPLC to confirm suspicious non-compliant samples according to Commission Regulation (EC) 1441/2007. Histamine concentration >2.5 mg kg−1 was detected in 58% of total samples with 5% of non-compliance. Among 216 samples of fresh fish, 70% had a content of histamine less than 10 mg kg−1, while a percentage of 6% showed histamine level higher than 100 mg kg−1. Although for processed anchovies a contamination below 50 mg kg−1 in 21 samples (75%) was observed, the highest percentage of “non-compliant” samples (14%) was found. Good quality of canned tuna and canned mackerel was demonstrated by histamine content lower than 10 mg kg−1. The validation of both ELISA and HPLC methods and the optimization of the confirmatory method were also reported.

Italy is dealing with iodine deficiency since ancient times. In 1848 an ad hoc committee appointed by the king of Sardinia, identified extensive areas afflicted by endemic goiter and endemic cretinism in Piedmont, Liguria and Sardinia. Since then many epidemiological studies have been conducted in our country. These showed that iodine deficiency was present not only in mountain areas but also in coastal areas. In 1972 the iodization of salt at 15 mg/kg was allowed by law and iodized salt was distributed on request to selected endemic areas. Five years later the distribution was extended to the whole country. However the sale of iodized salt was not mandatory at that time and only a small fraction of the Italian population started using iodized salt. In 1991 the content of iodine in the salt was raised to 30 mg/kg and in 2005 a nationwide salt iodization program was finally implemented. Some years later a nationwide monitoring program of iodine prophylaxis was also implemented. Since 2005 the sale of iodized salt in Italian supermarkets has increased (34% in 2006, 55% in 2012), although it has been observed that the use of iodized salt is still low in the communal eating areas and in the food industry. These data are coherent with recent epidemiological studies showing that some regions in our country are still characterized by mild iodine deficiency and a high frequency of goiter and other iodine deficiency disorders. This implies that further efforts should be made to successfully correct iodine deficiency in Italy.

Flavomycin complex is an antibiotic banned in the European Union as an additive in feed stuffs. As a consequence, the monitoring programmes for official control within the Community require analysis of feeds for possible illegal use of flavomycin. A method for unambiguous identification and quantification of moenomycin A, the main pharmacologically active component of flamomycin complex, in several feeds by liquid chromatography coupled to electrospray ion trap mass spectrometry (LC/ESI-MS/MS) is herein described for the first time. The method was developed to be used as a confirmative analytical tool for the network of Italian official control laboratories; both the singly and doubly charged molecular ions were observed as precursor ions, from which four product ions were selected for both quantitative analysis and unambiguous identification of moenomycin A. The method was in-house validated for feeds in the concentration range 0.50-30.0 microg/g, according to the Regulation 882/2004/EC requirements. Mean recoveries ranging between 83.9-94.2% and relative standard deviations

The analytical issues connected with chromium determination in foodstuffs by quadrupole inductively coupled plasma–mass spectrometry (Q-ICP-MS) were addressed, including signal stability, spectral interferences and the use of mathematical correction equations. The analytical performance was compared to that of electrothermal atomisation–atomic absorption spectrometry (ETA-AAS), selected as reference method. Five food certified reference materials (CRMs), including two meat-based CRMs not previously characterized for their Cr content, were included in the study. The use of ultrasonic nebulization (UN) and the adoption of 53Cr as analytical mass allowed precise and accurate results to be obtained by Q-ICP-MS, with lower detection limits than ETA-AAS.

The presence of cadmium (Cd), lead (Pb) and mercury (Hg) was investigated in fish and seafood products collected from the FAO Major Fishing Area 34, Eastern Central Atlantic. Samples were purchased from different retail outlets in Italy. Samples were selected so as to assess human exposure through diet. Metals were detected by Q-ICP-MS and Hg-AAS. All the metal concentrations detected were largely below the maximum levels (MLs) established by the European Union. The exposure assessment was undertaken by matching the concentration of Cd, Pb and total Hg in fish and other seafood products selected purposed according to Italian consumption data. The estimated weekly intakes (EWIs) for the evaluated elements related to the consumption of fish and other seafood products by the median of the Italian total population accounted for 14%, 2% and 14% of the standard tolerable weekly intake (TWI) for Cd and Hg as well as the provisional tolerable weekly intake (PTWI) for Pb, respectively.

Ultra performance liquid chromatography (UPLC) hyphenated to tandem mass spectrometry (MS/MS) was used for the development of an analytical method capable of simultaneous identification and quantification of 18 β -agonist compounds namely, brombuterol, chlorbrombuterol, cimaterol, cimbuterol, clenbuterol, clencyclohexerol, clenisopenterol, clenpenterol, clenproperol, hydroxymetylclenbuterol, isoxsuprine, mabuterol, mapenterol (clenbuterol-like compounds), ractopamine, ritodrine, salbutamol, salmeterol (salbutamol-like compounds) and zilpaterol in bovine urine. In compliance with the Commission Decision 2002/657/EC (CD 2002/657/EC), the method was validated applying a matrix-comprehensive in-house validation approach based on a fractional factorial design. Six experimental factors varied on two levels were selected for this purpose. The relevant validation parameters such as decision limit CCα (range, 0.24–0.51 μg L − 1 ) and detection capability CCβ (range, 0.27–0.71 μg L − 1 ), within-laboratory reproducibility ( β -agonist compounds in the framework of official monitoring plans.

Salt iodization is one of the preferred strategies to eradicate iodine-deficiency disorders worldwide. The aim of this study was (i) to estimate the daily iodine intake in the Italian population, (ii) to verify whether the recently recommended salt intake of 5 g/day in adults and less in children may be sufficient to achieve the adequate daily iodine intake, (iii) to estimate the contribution of iodine-enriched vegetables to the total daily iodine intake.The content of iodine has been evaluated in 288 food samples highly representative of Italian eating habits. The contribution of foods and iodized salt to the total daily iodine intake was estimated in adults, adolescents, and children on the basis of available nationwide data of food consumption.The total daily iodine intake owing to food and uniodized salt consumption resulted lower than the daily requirement in all age groups. With a daily consumption of 5 g of salt iodized at 30 mg/kg, the estimated daily iodine intake resulted within the range of optimal iodine intake in all age groups. In children the recommended iodine intake is achieved with a daily consumption of 3 g of iodized salt. Moreover, the daily iodine intake significantly increased in all age groups if the consumption of iodine-enriched vegetables was considered in substitution of non iodine-enriched ones.Our study suggests that the recommended quantities of salt, if iodized at 30 mg/kg, are sufficient to achieve the adequate daily iodine intake both in adults and children.

The cadmium content of muscle, liver, kidney and blood samples from 62 horses slaughtered in Italy was investigated. Cadmium was determined by graphite furnace atomic absorption spectrometry (GFAAS) after wet digestion of the samples. The mean and median contents of all samples were (on a fresh weight basis) 75 and 41 micrograms kg-1 for muscle, 2.46 and 2.10 mg kg-1 for liver, 20.0 and 13.5 mg kg-1 for kidney. The cadmium level in blood samples was always below 6 micrograms l-1. The cadmium concentrations in muscle, liver and kidney were found to be related to the life span of the specimens and increased with age. Females exhibited higher levels than males, but this difference was significant only in the case of muscle tissue. The geographical origin was recognized as the main factor influencing the cadmium content of the equine specimens analysed. The differences between horses coming from the three main breeding countries considered in this study (Poland, Lithuania, Hungary) were marked and statistically significant. The average intake of cadmium from equine meat was estimated for the general population and for population groups resident in areas with high consumption of this food item. While the cadmium intake from equine meat for the average consumer accounts for about 1% of the total cadmium intake estimated for Italy, in the latter case the enhanced consumption of equine muscle is often accompanied with the consumption of substantial quantities of equine liver and this may lead to high cadmium intakes.

Methyleugenol (ME) is a natural constituent of the essential oils of a number of plants widely used in foodstuffs as flavouring agents. Its occurrence, routes of intake, and available information on toxicity, kinetics and metabolism are reviewed. In view of the carcinogenic potential of ME, the need to check its presence in food products with effective analytical methods is pointed out.

The purpose of this study was to estimate the lead intake from crystalware resulting from short-term contacts with beverages, under conditions that are likely to occur to a consumer. The extraction ability of different kinds of beverages was estimated by comparison with 4% acetic acid under conditions of continuous contact for 3 h. It was found that lead release increased in the following order: cola drinkHAcwhiskywhite wine. Under conditions of repeated use under different scenarios, lead release showed a steep decrease with increasing number of contacts, for both wine and cola drink. The maximum lead intake resulted from the cola drink, corresponding to an ingestion of 14.5 micrograms Pb for consumption of 350 ml beverage. Assuming a fixed contribution from the diet of 71 micrograms/day, in the six scenarios taken into consideration, total daily lead intake levels ranged from a minimum of about 76 micrograms up to a maximum of 86 micrograms lead. As these values, converted on a weekly basis, would correspond to 35% and 40% PTWI respectively, significant health risks resulting from the ingestion of beverages in contact with crystalware can be excluded. Finally it was found that the use of a dishwater did not affect significantly the release of lead into wine, while release into cola drink was slightly but significantly increased after the third cycle.

Cd bioaccumulation pattern was investigated in Mediterranean spider crab ( Maya squinado , Herbst, 1788) collected from the northern Adriatic Sea. Specimens were caught in the framework of a monitoring plan in order to quantify the Cd distribution into different organs and tissues of crab. For this purpose, Cd level was studied in appendages, cephalothorax, abdomen as well as gonads. Cd concentrations were found largely below the Maximum Level (ML) established at the European Union (EU) level for muscle from crab appendages (found mean 0.011 mg kg −1 ) and approximately amounted to 2% of the EU ML (0.50 mg kg −1 ). The higher Cd concentrations were found in organs and tissues included in crab body such as abdomen, chephalotorax and gonads with respect to appendages. Chephalotorax showed the highest metal concentration (mean value of 1.19 mg kg −1 ). The possible differences in Cd bioaccumulation rate among crab organs and tissues were also investigated applying a parametric linear regression. A major Cd bioaccumulation rate was revealed in chephalotorax with respect to other analyzed organs and tissues. Furthermore, the evaluation of health risk related to human consumption of the Mediterranean spider crab has been studied for median of total population, median and 95th percentile of consumers of Italy. The observed results highlighted that the consumption of organs and tissues included in crab body such as abdomen, gonads and, in particular, chephalotorax substantially increased the Cd intake reaching also alarming Estimated Weekly Intake (EWI) values especially for median and 95th percentile of Italian consumers.

Iron homeostasis in humans is tightly regulated by mechanisms aimed to conserve iron for reutilisation, with a negligible role played by excretory mechanisms. In a previous study we found that mice have an astonishing ability to tolerate very high doses of parenterally administered iron dextran. Whether this ability is linked to the existence of an excretory pathway remains to be ascertained.Iron overload was generated by intraperitoneal injections of iron dextran (1 g/kg) administered once a week for 8 weeks in two different mouse strains (C57bl/6 and B6D2F1). Urinary and faecal iron excretion was assessed by inductively coupling plasma-mass spectrometry, whereas cardiac and liver architecture was evaluated by echocardiography and histological methods. For both strains, 24-hour faeces and urine samples were collected and iron concentration was determined on days 0, 1 and 2 after iron administration.In iron-overloaded C57bl/6 mice, the faecal iron concentration increased by 218% and 157% on days 1 and 2, respectively (p0.01). The iron excreted represented a loss of 14% of total iron administered. Similar but smaller changes was also found in B6D2F1 mice. Conversely, we found no significant changes in the concentration of iron in the urine in either of the strains of mice. In both strains, histological examination showed accumulation of iron in the liver and heart which tended to decrease over time.This study indicates that mice have a mechanism for removal of excess body iron and provides insights into the possible mechanisms of excretion.

The presence of selected toxic heavy metals, such as cadmium (Cd), lead (Pb) and mercury (Hg), was investigated in fish and seafood products, namely, blue mussel, carpet shell clam, European squid, veined squid, deep-water rose shrimp, red mullet, European seabass, gilthead seabream, Atlantic cod, European hake, Atlantic bluefin tuna and swordfish so as to assess their human exposure through diet. Metals were detected by quadrupole inductively coupled plasma mass spectrometry (Q-ICP-MS) and hydride generation atomic absorption spectrometry (Hg-AAS). Measurements of Cd, Pb and Hg were performed by means of analytical methods validated in compliance with UNI CEI EN ISO/IEC 17025 [2005. General requirements for the competence of testing and calibration laboratories. Milano (Italy): UNI Ente Nazionale Italiano di Unificazione]. The exposure assessment was undertaken matching the levels of Cd, Pb and total Hg with consumption data related to fish and seafood products selected for this purpose. In order to establish human health implications, the estimated weekly intakes (EWIs) for Cd, Pb and Hg were compared with the standard tolerable weekly intakes (TWI) for Cd and provisional tolerable weekly intakes (PTWIs) for Pb and Hg stipulated by the European Food Safety Authority (EFSA) and the Food and Agriculture Organization/World Health Organization (FAO/WHO) Joint Expert Committee on Food Additives (JECFA). The found metal concentrations were largely below the maximum levels (MLs) established at the European Union level with the exception of Cd. This metal exceeded the MLs in squid, red mullet, European hake and Atlantic cod. Squid and blue mussel showed the highest Pb concentrations which accounted for 60% and 10% of the MLs, respectively. Highest Hg levels were found in predatory fish. The concentrations of Hg in swordfish, Atlantic bluefin tuna and red mullet accounted for 50%, 30% and 30% of the MLs, respectively. The EWIs for Cd, Pb and Hg related to the consumption of fish and seafood products by the median of the Italian total population accounted for 20%, 1.5% and 10% of the standard TWI for Cd as well as PTWIs for Pb and Hg, respectively. Furthermore, the EWIs estimated using consumption data concerning Italian consumers did not exceed the standard TWI and PTWIs, except for Cd at 95th percentile.

Flavomycin complex is an antibiotic banned in the European Union as an additive in feed stuffs. As a consequence, the monitoring programmes for official control within the Community require analysis of feeds for possible illegal use of flavomycin. A method for unambiguous identification and quantification of moenomycin A, the main pharmacologically active component of flamomycin complex, in several feeds by liquid chromatography coupled to electrospray ion trap mass spectrometry (LC/ESI-MS/MS) is herein described for the first time. The method was developed to be used as a confirmative analytical tool for the network of Italian official control laboratories; both the singly and doubly charged molecular ions were observed as precursor ions, from which four product ions were selected for both quantitative analysis and unambiguous identification of moenomycin A. The method was in-house validated for feeds in the concentration range 0.50-30.0 microg/g, according to the Regulation 882/2004/EC requirements. Mean recoveries ranging between 83.9-94.2% and relative standard deviations23% account for method trueness and repeatability, respectively. Moreover, other analytical performance parameters, i.e. method specificity, ruggedness, the linearity of detector response, the limit of quantification (LOQ), the limit of detection (LOD), and measurement uncertainty were evaluated and reported. The ion trap LC/ESI-MS/MS method is highly selective and reliable; high drug recovery, good reproducibility and an LOQ down to 0.10 microg/g guarantee its applicability for confirmatory purposes in the official control activity in Italy.

Sample contamination as a consequence of abrasion of grinding tools during the homogenization of food materials to be analysed for trace elements was addressed. The possible release of 15 trace elements (Al, As, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Se, Sn, V, Zn) from six different grinding and milling devices, operating either continuously or discontinuously, was evaluated. All the devices were commercially available and were representative of models usually employed in food and agricultural laboratories. Wheat grains belonging to one soft and one durum cultivar were used as test material. The determination of the analyte concentrations in subsamples submitted to the different preparation treatments was performed by quadrupole inductively coupled plasma mass spectrometry (Q-ICP-MS). Accordingly, a suitable digestion method was developed and the ArC+ interference affecting Cr determination was evaluated and corrected. Statistical differences with respect to the control were detected for 10 elements (Al, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb) and in most cases contamination of the samples was traced back to the composition of the grinding equipment. None of the investigated devices was contamination-free with respect to all of the quantified elements. Abrasion of the grinding tools was higher with durum wheat than with soft wheat as a consequence of their different hardiness.

Studies were performed on the efficacy, residues and in vitro enterocyte toxicity of 4-hexylresorcinol (4-HR), which could be utilized as an inhibitor of shrimp melanosis (black spot). Mediterranean sea shrimp (Parapaeneus longirostris) were treated with solutions of 4-HR in sea-water, at three different concentrations, 25, 50 or 100 mg/kg of shrimp, to test its antioxidative property. As a comparison a group of shrimp was treated with sodium metabisulphite (1 g/kg), while another group was left untreated. 4-HR showed a marked ability to inhibit or slow down melanosis (black spot) in shrimp; the most effective concentration was 100 mg/kg within an optimum period of 7 days but with effects up to the tenth day. During the first 5 days, 4-HR residues in the edible part of the shrimp showed a fast decrease in all three groups, going from initial average values of 20 mg/kg at 0 time, to 0.9 in the group treated at 25 mg/kg; from 42 to 1.8 mg/kg in the group at 50 mg/kg and from 85 to 1.9 mg/kg in the group at 100 mg/kg. In vitro studies on enterocyte-like Caco-2 cells did not indicate any cytotoxic effect up to a concentration of 50 micrograms/ml. Moreover, no inhibition of protein synthesis was observed, which lends further support to the absence of significant damage to the intestinal mucosa induced by 4-HR. The available database on 4-HR pharmacology and toxicology is inadequate to determine even a provisional ADI. There is negative evidence of carcinogenesis and no significant untoward effects were observed in humans when it was used as an anthelmintic. However, it is not possible to determine a NOEL for non-genotoxic effects. 4-HR could become an interesting alternative to the use of sulphites to prevent black spot. However, a more complete database is needed to achieve a regulatory evaluation.