Risk identification in food safety: Strategy and outcomes of the EFSA emerging risks exchange network (EREN), 2010–2014

The European Food Safety Authority (EFSA) established an Emerging Risks Exchange Network (EREN) to exchange information between EFSA and the Member states (MSs) on possible emerging risks for food and feed safety in 2010. The Network is composed of delegates from MSs and Norway designated through the Advisory Forum of EFSA and observers from the European Commission, EU pre-accession countries, the Food and Drug Administration of the USA and the Food and Agricultural Organisation of the United Nations. Through 2010 to 2014, the EREN met 12 times. The EREN discussed a total of 63 signals of potential emerging issues that were presented and assessed using a standard template developed by the Emerging Risks unit of EFSA (EMRISK). Out of these signals, 39 originated from EFSA, 24 from MSs. The issues discussed were mainly microbiological and chemical hazards, but also food safety issues as result of illegal activity, new consumer consumption trends, biotoxins, new technologies and processes, allergens, animal health, environmental pollution, new analytical methods, new food packaging technology and unknown hazards were on the agenda. Based on the available evidence, EREN recommended whether an issue should be considered emerging or not, and if it merited further consideration, such as generating data on the issue, starting a full risk assessment and/or consultation of other bodies. According to the emerging risks identification process set in place at EFSA, the issues discussed and found of relevance by EREN were sent to the EFSA's Scientific Committee Standing Working Group on Emerging Risks for final evaluation. With four case studies, i.e the zoonotic potential of Usutu virus, risk of ciguatera fish poisoning in EU, zoonotic aspects of illegally imported wildlife products and benefits and risks of 3D food printing, the method developed to preliminary assess signals of potential emerging issues is presented and discussed.     

Three years of monitoring of PCDD/F,DL-PCB and NDL-PCB residues in bovine milk from Lombardy and Emilia Romagna regions (Italy): Contamination levels and human exposure assessment

In the three-years period 2012–2014, 160 cow milk samples from farms located in Lombardy and Emilia Romagna regions (Italy) were analyzed during the implementation of the Italian National Residues Monitoring Plan to assess the presence of PCDD/F, DL-PCB and NDL-PCB residues. The obtained contamination data were combined with cow milk consumption data from the Italian national dietary survey to estimate PCDD/F, DL-PCB and NDL-PCB human dietary exposure through the consumption of whole, semi skimmed and skimmed bovine milk. The exposure assessment was carried out separately for children, teenagers, adults and elderly. Average contamination levels of the analyzed samples were found to be 1.26 pg WHO-TEQ/g fat for the sum of PCDD/Fs and DL-PCBs and 9.30 ng/g fat for the sum of the 6 NDL-PCB indicators. PCB 126 was found to be the main contributor to the total WHO-TEQ. Using the upper bound approach, the estimated mean dietary intakes ranged from 0.07 pg WHO-TEQ/kg bw per day to 0.39 pg WHO-TEQ/kg bw per day for the sum of PCDD/Fs and DL-PCBs and considering exposure from whole milk. NDL-PCB mean dietary intakes resulted between 0.52 ng/kg bw per day and 2.86 ng/kg bw per day for consumption of whole milk. Children and teenagers were found to be the most exposed groups. This is the first time that Italian consumers exposure to NDL-PCBs is assessed using contamination data of cow milk produced in Italy.     

Antibiotic and synthetic growth promoters in animal diets: Review of impact and analytical methods

Food quality and safety have been a significant and pressing issue in recent years. In light of the FAO's definition of food security – the physical, social and economic access to sufficient and nutritious food – food safety plays a fundamental role. Animal feed and feeding is pivotal to the livestock industry, but the use of veterinary antibiotics (VAs) and synthetic growth promoters (SGP) diminishes the sustainability of the diets and can cause an accumulation of residues in animals (meat, milk and eggs) and the environment (water and soil pollution). Wastewater systems are another major pathway through which antibiotics and hormones can enter the environment, with negative consequences. In order to protect the planet through more sustainable feeding, the reduction of antibiotics and synthetic growth promoters is a key aim, in particular with the goal of reducing antibiotic resistance and allergies. Analytical methods play a crucial role in food analysis, to determine the presence of antibiotics and other additives. Recent methods are based on liquid chromatography with ultraviolet, fluorescence, or mass spectrophotometry detection, which is recognized as an essential technique in food analysis, able to identify more than 300 compounds in feed samples. In general, a monitoring program put in place to educate the population on the hazards of residues in animal products is necessary, in conjunction with a continuous decrease in the use of antibiotics and synthetic growth promoters in animal diets.     

Occurrence and molecular identification of anisakid nematodes isolated from Pacific cod (Gadus macrocephalus) caught off Korea

Anisakids nematodes from Pacific cod (Gadus macrocephalus) in Korea were investigated and their molecular identification was conducted, to assess the epidemiological role of Pacific cod in human anisakidosis in Korea. Totally 238 Pacific cod were caught from 5 different areas around Korean peninsula. Fish were dissected and carefully examined for collecting nematodes. PCR-RFLP and the subsequent sequencing were conducted for molecular identification of those nematodes. A high prevalence of infection (193/238, 81.1%) in Pacific cod was observed, and 1694 nematodes were collected. 79.1% (1340/1694) of the nematodes were found freely in the body cavity of Pacific cod, and the rest of them (20.9%, 354/1694) were in the digestive tract or attached to other organs. PCR-RFLP analysis using HinfI and RsaI restriction enzymes revealed 3 different banding patterns corresponding to Anisakis pegreffii, Hysterothylacium aduncum and hybrid genotype (Anisakis simplex × A. pegreffii), respectively. Of 1694 nematodes, 1280 (75.6%) were identified as A. pegreffii and 406 (24.0%) were H. aduncum. A. pegreffii occupied 84.0% (1125/1340) of the nematodes in the body cavity and 40.0% (132/330) of them in the digestive tract, but no nematodes were found in Pacific cod muscles.     

Aflatoxin contamination in unrecorded beers from Kenya – A health risk beyond ethanol

Samples of unrecorded opaque beers (n = 58; 40 based on maize, 5 on sorghum and 13 on other plants) and recorded wines (n = 8) in Kenya were screened for aflatoxins using a rapid ELISA technique followed by confirmation using liquid chromatography-tandem mass spectrometry. Six of the maize beers were obtained from Kibera slums in Nairobi County. Aflatoxin contamination was detected in six unrecorded beers (10%), but in none of the recorded wines. Remarkably, three of the aflatoxin positive samples were from the Kibera slums.The mean concentration of aflatoxins in the positive samples was 3.5 μg/L (range 1.8–6.8 μg/L), corresponding for an average consumption of 500 mL (1 standard drink) to a margin of exposure (MOE) of 36 (range: 15–58), which is considered as ‘risk’. On the other hand, the alcoholic strength of the aflatoxin positive samples had a mean of 4.3% vol (range 3.5–4.8%) corresponding to a MOE of 2.5 (range of 2.2–3.0) for the equivalent consumption volume. While aflatoxins pose a risk to the consumer, this risk is about 10 times lower than the risk of ethanol.The Joint FAO/WHO Expert Committee on Food Additives sets no acceptable daily intake for aflatoxins since they are genotoxic carcinogens and instead recommends for the reduction of aflatoxin dietary exposure as an important public health goal, particularly in populations who consume high levels of any potentially aflatoxins-contaminated food. Nevertheless, ethanol still posed a considerably higher risk in the unrecorded beers examined. However, consumers should be informed about aflatoxins, as these are an involuntary and unknown risk to them. In addition, producers should be educated about measures to reduce aflatoxins in alcoholic beverages.     

Determination,content analysis and removal efficiency of fining agents on ochratoxin A in Chinese wines

A simple and convenient HPLC-FD detection method for ochratoxin A (OTA) with a high detection limit and a short run time has been developed. OTA has been found in most samples of Chinese market wine, including domestic and imported wines, but the content was not very high. Only a few wines showed an OTA content that exceeded the EC and OIV limits, indicating that most Chinese market wines were safe. The OTA intake for Chinese from wine was 0.017 ng/kg (bw) per day, which was lower than the SCF and JECFA limits and also lower than in many other countries. This was mainly due to the low per capita wine consumption in China, but it is still necessary for the Chinese government and wine makers to monitor OTA levels in wine and to establish relevant regulations. An egg white treatment (0.20 mg/mL, 48 h) was the best removal method for OTA.     

The effects of tangerine peel (Citri reticulatae pericarpium) essential oils as glazing layer on freshness preservation of bream (Megalobrama amblycephala) during superchilling storage

This work was aim to the application of tangerine peel essential oils (TPEOs) as glazing layer on fish preservation. In this paper, essential oils were respectively extracted from the peel of ponkan, bitter orange and sweet orange. Their compositions were analyzed using GC–MS method. Fresh sample of bream was immersed into different TPEO for the formation of glazing layers on the surface and stored at −1 ±0.2 °C for 25-days storage. A thorough study of freshness evaluation was carried out and statistics analysis was performed to analyze the effect of TPEO. The result showed that the glazing layers of TPEO can effectively slow down the degradation process of fish samples and the resulting variations in electrical, moisture, chemical, microbial, sensory and textural characteristics. Additionally, statistical analysis showed that there are significant (most of p is less than 0.05) difference between control and treated sample, but insignificant (most of p is more than 0.05) differences among the samples with glazing layer of different TPEOs.The work indicated TPEOs had remarkable effect in the storage of aquatic products.     

Detection of selected corticosteroids and anabolic steroids in calf milk replacers by liquid chromatography–electrospray ionisation – Tandem mass spectrometry

The use of corticosteroids and anabolic steroids in food producing animals is regulated or banned in the European Union (EU). However, their use as growth promoters cannot be excluded. Milk replacers, considered by EU legislation as feeds, may be a good way of administration of these compounds. In order to improve the control of growth promoter utilization in animal husbandry and preventing possible consequences to animal welfare, we developed a method for multiresidue analysis of prednisolone, prednisone, dexamethasone, cortisone, cortisol, 17α- and 17β-boldenone and their precursor androstadienedione (ADD), testosterone, epitestosterone, 17α- and 17β-nandrolone, and trenbolone in powdered milk for calves. All analytes were extracted, after a common deproteinization and defatting sample pre-treatment, by a unique immunoaffinity column and analysed by liquid chromatography tandem mass spectrometry (LC–MS/MS) in both positive and negative electrospray ionization (ESI) modes. The method was validated according to the criteria of the Commission Decision 2002/657/CE. The analytical limits were from 0.39 to 0.73 ng mL⁻¹ for the decision limit (CCα) and 0.46–0.99 ng mL⁻¹ for detection capability (CCβ). The analysis of 50 samples of milk replacers for calves, always revealed the presence of cortisol and cortisone (average concentrations 2.56 and 1.06 ng mL⁻¹, respectively), frequently testosterone and epitestosterone (1.24 and 0.63 ng mL⁻¹, respectively), occasionally β-nandrolone (0.82 ng mL⁻¹) and prednisolone (0.41 ng mL⁻¹). The other anabolic steroids were never found.     

Verifying the red wines adulteration through isotopic and chromatographic investigations coupled with multivariate statistic interpretation of the data

The assessment of wine traceability and authenticity is a major concern that has gained a lot of interest internationally since the wine has always been subjected to various fraudulent practices. Practiced since ancient times, wine fraud has become more sophisticated in the present day, taking many forms. Consumers, regulatory bodies and manufacturers are all interested to have reliable analytical tools and information to allow the authentication and detection of wine adulteration or incorrect labelling. This research study evaluates and proposes a possible strategy for the detection of adulterated sweet or medium sweet red table wines using appropriate chemical parameters which can reveal prohibited practices in the winemaking process. The work is performed on 29 table wine samples, bought from the market, packed in PET bottles. Exogenous addition of sugar and water in the counterfeited table red wines was detected by the measurement of stable isotopes content (δ¹³C and δ¹⁸O) known as origin markers and supplementary confirmed by other classical parameters, as the alcoholic strength of the wines (‰ vol.) and the presence of 5-(hydroxymethyl)-2-furaldehyde (HMF) and of synthetic sweeteners or synthetic red dyes used to correct deficiencies of taste and colour. Additionally, the nature and profile of anthocyanins, as indicator of the red colour of wine, was investigated in the table wines and then compared with that of authentic wines obtained by microvinification of Vitis vinifera, in order to determine their authenticity.     

Chemical taste taint accumulation in RAS farmed fish – A Fr 13 risk assessment demonstrated with geosmin (GSM) and 2-methylisoborneol (MIB) in barramundi (Lates calcarifer)

Recirculating Aquaculture System (RAS) farming of fish is becoming widespread. However, taste taint accumulation in the fish flesh as geosmin (GSM) and/or 2-methylisoborneol (MIB) is a major concern for farmers and consumers. Here we apply the emerging Fr 13 risk methodology (Food Control 50 (2015) 770–777; Food Control 29 (2013) 248–254) to demonstrate quantitatively the impact of naturally occurring fluctuations in these chemicals in RAS water on their accumulation in fish flesh. The approach is based on the time dependent accumulation model of Hathurusingha and Davey (Ecological Modelling 291 (2014) 242–249) in which taint chemicals enter the flesh via the gills and are diluted through metabolism and growth, together with a risk factor (p) such that for all p > 0 the chemical taint is above a desired threshold concentration (which includes a practical tolerance) respectively, 0.814 and 0.77, μg kg⁻¹ for GSM and MIB. Monte Carlo (with Latin Hypercube) sampling of chemical in the growth water (C_W), water temperature (T) and growth time (t) was used to simulate practical RAS farmed barramundi (Lates calcarifer), a premium fish, for up to 260 days growth. Results show some 10.10% of all harvests over the long term will result in fish with taste taint as GSM above the threshold concentration due to natural fluctuations in the RAS environment. For MIB this failure rate is 10.56%. The vulnerability to taste taint failure is shown to be impacted highly significantly by the time to harvest, and to a lesser extent by concentration and fluctuation of the taint chemicals in the RAS water. An advantage is growth time can be readily controlled by farmers; this should not exceed 240 day. The methodology appears generalizable and therefore applicable to a range of RAS farmed foods. Findings will be of immediate benefit to RAS farmers and to risk researchers in foods processing.     

Occurrence and co-occurrence of Fusarium mycotoxins in wheat grains and wheat flour from Romania

In this study, the presence of fourteen Fusarium mycotoxins, legislated by the European Union – deoxynivalenol, zearalenone, HT-2 and T-2 toxins (EC/1881/2006; 2013/165/EU), or non-legislated (five trichothecens and five “emerging” mycotoxins), was evaluated in 31 whole unprocessed wheat samples and 35 white wheat flour samples from different areas of Romania. For this purpose, a validated multi-mycotoxins liquid chromatography tandem mass spectrometry method was applied. Seventy three percent of the analyzed samples contained at least one mycotoxin. The highest occurrence was for enniatin B, 71% of the analyzed samples being positive (21–407 μg kg⁻¹). Regarding the legislated mycotoxins, deoxynivalenol was detected in 14% (111–1787 μg kg⁻¹) of the samples, while zearalenone was detected in 9% (51–1135 μg kg⁻¹). Only one sample was positive for neosolaniol. Concerning the co-occurrence, 42% of the samples were contaminated with two to five mycotoxins, the most frequent being the binary or tertiary combinations of enniatins. This is the first study applied to Romanian wheat grains and flour samples using a high sensitive multi-mycotoxins method, and which included also “emerging” mycotoxins.     

Minerals in vitro bioaccessibility and changes in textural and structural characteristics of uncooked pre-germinated brown rice influenced by ultra-high pressure

Ultra-high pressure (UHP) is considered as an effective processing method to enhance micronutrients utilization efficiency in germinated brown rice (GBR), but there is little information available for the effects of UHP treatments on mineral bioaccessibility and related structural characteristics. Therefore, this work examined the changes induced by UHP in the in vitro bioaccessibility of selected minerals and structural and textural properties at pressures, as well as their potential relationship. UHP treatments were applied at 100, 300 and 500 MPa for 10 min, and 0.1 MPa as control. Proximate analysis showed that free fatty acids (FFA) and moisture in GBR greatly increased after UHP treatment. Three representative minerals with different levels were chosen to assess the bioaccessibility, including phosphorus (P), magnesium (Mg) and manganese (Mn). The results demonstrated that the minerals bioaccessibility, measured by percentage solubility, were the highest when at 300 MPa whereas it decreased at 500 MPa. Correspondingly, microstructure imaging by scanning electron microscope (SEM) showed that the gelatinization of starch granule occurred at 300 MPa and it became more obvious when increased to 500 MPa. Starch gelatinization significantly lowered the hardness of GBR grains, simultaneously accompanied with the decrease of cohesiveness, gumminess and resilience through texture profile analysis (TPA). Multivariate analysis by principal component analysis and canonical correspondence analysis indicated a highly intra-associated (p < 0.01) TPA parameters and a close correlation between the bioaccessibility and hardness, chewiness and springiness. These results provide information on minerals bioaccessibility and structural properties of uncooked GBR after UHP treatment, which could lay the foundation for further elucidating the correlation between structure and micronutrients bioaccessibility.     

A Simple Colorimetric Enzymatic-Assay,based on immobilization of acetylcholinesterase by adsorption,for sensitive detection of organophosphorus insecticides in olive oil

Over the last decades, cholinesterase (ChE) biosensors/bioassays have emerged as an ultra sensitive and rapid technique for pesticides detection in environmental monitoring, food and quality control. In this work a Simple Colorimetric Enzymatic-Assay for the sensitive detection of three organophosphorus insecticides widely used in the treatment of olive trees: Malathion, Dimethoate and Methidathion in their oxidized form was developed. The systems are based on the immobilization of 0.3 mIU of electric eel acetylcholinesterase by adsorption on MaxiSorp™ microplates, and tested using standard solutions and real samples of olive oil. The developed colorimetric tests showed good analytical performances with limits of detection as low as 10⁻⁹ M for the widely used pesticide malaoxon (oxidized malathion), as well as a good reproducibility, good operational and storage stability and they can be applied directly on olive oil real samples without a laborious pre-treatment and after a simple liquid–liquid extraction, which demonstrate the potentiality of this technique to be used for commercial purpose.     

Effect of nutmeg (Myristica fragrans) essential oil on the oxidative and microbial stability of cooked sausage during refrigerated storage

This paper examines the oxidative and microbial stability of cooked sausages, produced with the addition of 10 ppm (NO1) and 20 ppm (NO2) nutmeg (Myristica fragrans) essential oil. Instrumental parameters of color (CIE L*, CIE a* and CIE b*), Thiobarbituric acid-reactive substance (TBARS) values, microbial profile and sensory properties of aroma have been determined on the 1st, 30th, 45th, and 60th day of storage. Addition of the nutmeg essential oil had no effect on the color of cooked sausages. At the end of the storage, NO2 sausages had the best oxidative and microbial stability. TBARS values of NO1 and NO2 sausages were 1.21 mg MDA/kg and 0.95 mg MDA/kg, respectively, and were significantly lower (P < 0.05) compared to control (1.53 mg MDA/kg). Total number of aerobic mesophilic bacteria was lowest in NO2 sausages (78.3 cfu/g) and highest in control (185 cfu/g). After 45 and 60 days of storage, sensory properties of aroma of NO1 (4.21; 3.92) and NO2 sausages (4.39, 4.28) were better compared to those in control (4.07, 3.25). Hence, the addition of nutmeg essential oil in amount of 20 ppm can be successfully applied in order to extend the shelf life of cooked sausages.     

Antimicrobial effects of cinnamon bark oil on microbial composition and quality of grass carp (Ctenopharyngodon idellus) fillets during chilled storage

In this study, 11 essential oils were initially screened for antimicrobial activity against Aeromonas veronii, Acinetobacter johnsonii, Shewanella putrefaciens, and Pseudomonas jessenii. Cinnamon bark oil exhibited the highest antimicrobial activity and was used as a natural preservative in grass carp fillets stored at 4 ± 1 °C. Sensory assessment, total volatile basic nitrogen (TVB-N), color, biogenic amines, adenosine triphosphate (ATP)-related compounds, K-value, and presence of microbiota were analyzed during storage. Cinnamon bark oil treatment was found to be effective in enhancing organoleptic quality, inhibiting microbial growth, and delaying the increase of TVB-N, putrescine, cadaverine and K-value. Based on sensory analysis, shelf-life of grass carp fillets was 8 days for the control and 12 days for the treatment group. According to high-throughput sequencing, Aeromonas and Pseudomonas were the predominant microbiota in fresh grass carp. As storage time progressed, the microbial composition of both control and treatment samples became less diverse. Pseudomonas followed by Aeromonas were dominant in spoiled control samples. However, there were significant differences in microbial composition between control and treatment samples at the end of storage. Cinnamon bark oil treatment inhibited the growth of Aeromonas and Shewanella, and Pseudomonas was the only predominant microbiota found in spoiled treatment samples.     

Stable isotope and fatty acid compositions of monovarietal olive oils: Implications of ripening stage and climate effects as determinants in traceability studies

The quantitative variability of isotopic and fatty acid compositions in extra virgin olive oil was investigated in relation to variety, olive ripeness index and geographical origin. The study was carried out in two environments of Umbria region (central Italy), differing in annual/seasonal temperatures and rainfalls. Significant correlation was found between oil isotope compositions and climatic seasonal changes during fruit ripening. Moreover, significant isotopic differences among cultivars were related to the period of maximum oil accumulation in the fruits and to the fatty acid profile. A carbon kinetic isotopic effect could be associated with the fatty acid elongation from palmitic to stearic acid and to the unsaturation step leading from oleic to linoleic. No correlation was found between stable isotope and fatty acid compositions during fruit ripening. The analyzed cultivar and ripening effects on oil traceability issues are discussed in order to focus the resolution power of stable isotope methodologies and the possibility to distinguish monovarietal oils from the same area.     

Development of an HTS system to identify natural chemicals that specifically inhibit Escherichia coli O157:H7 adhesion to host cells

Adhesion to intestinal epithelium is critical for virulence and infection of Escherichia coli O157:H7, which is a major cause of foodborne illnesses. Targeting adhesion but not growth of the pathogen is an effective approach to control bacterial infections. In this study, a high-throughput screening system was developed to identify anti-adhesive natural chemicals using a reporter strain whose activity entirely depends on Ler since genes encoding the locus of enterocyte effacement (LEE) which forms attaching and effacing lesions are known to be activated by Ler. A selected hit, yomogin, suppressed the expression of virulent genes encoded within the LEE without affecting bacterial growth itself. Yomogin significantly reduced the adherence of E. coli O157:H7 to Caco-2 epithelial cells and cytotoxicity. These results indicated that the developed system allows for rapid and cost-effective discovery of novel anti-adhesive agents and yomogin can prevent early stage of E. coli O157:H7 pathogenesis.     

Bactericidal effect of various non-thermal plasma agents and the influence of experimental conditions in microbial inactivation: A review

Microbial inactivation using non-thermal gas discharge at atmospheric pressure has become a subject of significant research effort in the recent years. In this paper, we reviewed the different viewpoints proposed by various researchers, and discussed the reasons for arriving at these conclusions. We summarized some general rules, and offered a proposal to study the reasons behind their conclusions by building mathematical model for prediction of principle factors. The future prospects for the application of plasma are outlined.     

Effects of Salmonella bacteriophage,nisin and potassium sorbate and their combination on safety and shelf life of fresh chilled pork

Salmonella is an important foodborne pathogen and a serious threat to human health worldwide. This study was to reduce Salmonella and the spoilage bacteria on fresh chilled pork using bacteriophage, nisin, and potassium sorbate (PS) along with their combinations. Microbial, chemical, and sensory qualities of the fresh chilled pork (artificially contaminated with Salmonella 3 log CFU/g) treated with bacteriophage (9 log PFU/g), nisin (5000 IU/g), PS (2 mg/g) and their combinations were evaluated. The result showed that all the samples treated with phage could significantly (P < 0.05) reduce Salmonella population on fresh chilled pork. The combination treatment of nisin, PS and phage (N-PS-P) could significantly lower total viable counts (TVC), TVB-N and TBARS of the chilled pork during the storage period. The TVC of sample treated by N-PS-P was reduced by 2.3 log CFU/g at 7th day. It was also found through the electronic nose detection that the N-PS-P treatment was able to significantly reduce odour and maintain good sensory of the chilled pork. Hence, the N-PS-P treatment extended the shelf life of fresh chilled pork up to 14 days. No adverse effect of the phage on the chilled pork was observed. In conclusion, this study suggests that the phage and its combination with nisin and PS have great potential to be used as a good preservative for fresh chilled pork.