Investigation of the contents of biogenic amines and some food safety parameters of various commercially salted fish products

Seventy-eight salted fish products originating from Turkey and European countries were analyzed for their content of biogenic amines (BA), water activity (a_w), salt and moisture, and their microbiological counts. About 10% of the samples exceeded regulated histamine levels established by the FDA and/or EU authorities. The highest histamine value was found for brined anchovy as 422 ppm. Among the other BAs, the highest level was obtained for tyramine as 524 ppm. The low a_w (<0.9) values were obtained with samples containing water phase salt (%WPS) content over 20%. Brined and dry salted products had higher %WPS than lakerda, which is a Turkish and Greek salted fish product. Despite of salt used as preservative agent in fish processing, it is found that salting alone is not enough to guarantee low levels of histamine and other BAs. This study indicated that commercial salted fish products, particularly lakerda, may still present health risk for BAs depending on processing technique, handling and storage conditions of raw material, and during processing. Therefore, processors and regulatory bodies are advised to re-evaluate their productions for safe handling measures.     

Anisakis survival in refrigerated fish products under CO2 modified-atmosphere

With the aim to assess the survival of Anisakis infection in fish stored under CO₂ modified-atmosphere, third-stage larvae (L3) of Anisakis simplex were inoculated in samples of skinless fish musculature and then packaged in three different CO₂ modified-atmospheres commonly used to preserved fresh fish products. The microenvironment provided by CO₂ modified-atmosphere packaging (MAP) does not impair the well-being and somatic antigenic properties of Anisakis larvae in infected fresh fish. This result indicates that MAP does not protect against anisakiasis risk. Therefore proactive regular inspections in commercial fresh fish should be done by MAP operators in fishery industries to ensure that healthy products (i.e., parasite-free fish products) reach the consumer, thus accomplishing with the co-responsibility criterion established in the EU food hygiene legislation.     

DNA barcoding as a tool for detecting mislabeling of fishery products imported from third countries: An official survey conducted at the Border Inspection Post of Livorno-Pisa (Italy)

The importation of fishery products into the European Union (EU) is constantly rising. The aim of this study was to conduct a survey on labeling non-compliances on fishery products imported from extra-European countries, in collaboration with the veterinary staff of the Border Inspection Post of Livorno-Pisa (BIP), of the Italian Ministry of Health. The correspondence between the products' identity and the scientific denominations reported on the accompanying certificates was checked using the DNA barcoding method. Overall, 277 products belonging to different categories (fish, cephalopods, crustaceans, bivalves, amphibian) were submitted to analysis for species identification. The comparison of the molecular results with the scientific names declared on the accompanying documents showed that 22.5% (95%CI 17.8–28.0) of the analyzed products were mislabeled. The highest percentage of mislabeling was observed on cephalopod based products (43.8%, 95% CI 32.3–55.9), followed by crustaceans (17.0%, 95% CI 9.2–29.2) and fish (14.0%, 95% CI 8.7–21.9). A higher rate was found in products imported from China, Vietnam and Thailand. The present study is the first survey on mislabeling in products sampled at BIPs in Italy. The results highlight the need of implementing analytical checks, based on DNA analysis, on incoming fishery products.     

Development of a Catch-damage-index to assess the quality of cod at landing

Quality and quality management are of increasing concern and importance in all parts of the fishery chain. Fishing gears and handling onboard the vessel have multiple impacts on fish quality, and damages caused during these operations may affect the product quality considerably, resulting in downgrading of the end products. The European fish sector consider measurement of quality defects and other quality factors as important, but methods for systematic evaluation of quality defects caused during catching operations have not been published to any extent.In this work cod caught by different fishing gears commonly used in the Norwegian coastal fisheries were examined for catch-related damages at landing. Other species like saithe and haddock may suffer the same damages, but due to being the economically most important specie in Norway, cod is chosen as model. The severities of the damages are described, and the influence of the damages on the quality of cod products is considered. Based on this, a Catch-damage-index (CDI) scheme is elaborated.     

Comparison of IEF patterns of sarcoplasmic proteins of fish from North Atlantic and Aegean Sea

Identification of processed fish lacking morphological characteristics is of growing importance in the times of increasing global trade and illegal fishery. Protein electrophoresis techniques, like isoelectric focusing (IEF) of water-soluble proteins, are fast and inexpensive tools for identification of raw fishery products. Protein profiles of raw fish or fillet of the same species were found to be remarkably constant. However, it cannot be excluded that protein patterns of fish of the same species from distant stocks may show some variation.The protein profiles of commercially important fish from different fishing grounds were determined by collecting samples of 38 fish species from Aegean Sea and Turkish fish farms and of five species from North Atlantic waters. The raw fillet of the fish was analysed by IEF of water-soluble (sarcoplasmic) proteins. Species-specific protein profiles were obtained suited for identification of raw fish or fillet. When protein patterns and pI values were compared between Mediterranean and North Atlantic fish of the respective species, only minor differences were observed. These differences were related to the intensity of protein bands, but not to the position within the pH gradient of the IEF gel.The pI values (isoelectric points) of the prominent protein bands of the fish were determined to be used as a database.     

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.     

Food safety aspects of fresh Nile perch (Lates niloticus) fillets from Lake Victoria imported to the European market: Helminth parasites and microbiological status

Nile perch (Lates niloticus) is a popular, usually low priced food fish in many countries worldwide, and the most important export fish from Lake Victoria and its adjacent countries Uganda, Tanzania and Kenya. Despite its high import figures, the constantly increasing demand within the EU for ‘new’ fish species and the results of initial studies indicating high coliform bacterial loads, food safety aspects regarding its microbiological as well as parasitological status have been hitherto widely neglected. In the present study imported fresh fish fillets as well as superchilled fish from a German retail market and a Dutch wholesaler were examined to evaluate the current microbiological and parasitological quality. A total of 200 fresh fillets as well as 20 superchilled fish (beheaded, eviscerated, descaled) samples were screened for the presence of helminth parasites by visual inspection, candling, the UV-press method and artificial digestion method. The study includes as well a quantitative test on the recovery rate of metacercarial cysts. Microbial assessment was performed on ten fresh fillets and on the skin and tissue of 20 superchilled fish. Helminth parasites were neither detected in the edible parts nor in the skin of imported products. Results of the microbiological examination revealed low levels of bacterial contamination of superchilled fish samples, whereas high total viable counts and high levels of naturally present bacterial pathogens of the genus Aeromonas spp. and Pseudomonas spp. were identified from the fresh fillets. Our findings indicate that hygienic production conditions have improved in comparison to previous studies. As total viable counts as well as specific spoiling bacteria were high in fresh fillets, we recommend adequate heat treatment of fresh Nile perch fillets before consumption.     

A model for risk-based monitoring of contaminants in feed ingredients

A qualitative spreadsheet model has been developed for ranking feed ingredients on the basis of the potential risk of exceeding existing guidance or maximum levels in the EU for a certain contaminant, and the potential consequence of the presence of this contaminant on the health of animals and/or humans. The approach was based on the general concept of risk, being frequency times consequences of presence of the contaminant. Contamination of compound feeds due to presence of the contaminant in feed ingredients was estimated, per animal category, by: annual volumes of feed ingredients used for feed production, stratified per country of origin; the portion of each ingredient in compound feed formulations used for various animal categories; and the potential contamination of an ingredient per country of origin. The consequences of the contamination were accounted for by two consequence factors, both estimated per animal category: one for the potential impact of the contaminant on the health of the target animal, and one for the impact on human health, related to the possible formation of residues in animal derived food products.The use of the model was demonstrated by its application to the presence of dioxins and dl-PCBs in compound feed for farm animals produced in the Netherlands in 2013 and 2014. Model results include the relative contribution, based on relative ranking scores, of each feed ingredient to the chance of exceeding limits and potential consequences on animal and human health. Feed ingredients ranking highest were palm oil, other fats and oils, dried products like bakery products, sunflower expeller/extracted, maize, and fish meal.The model can be used by risk managers in feed industry and by governmental bodies for supporting decision making on the optimal allocation of resources for control of ingredients for compound feed production for presence of contaminants.     

Employing visual inspection and Magnetic Resonance Imaging to investigate Anisakis simplex s.l. infection in herring viscera

Anisakis spp. and Pseudoterranova spp. are anisakid nematodes with complex life cycles that utilise crustacean, fish and, finally, marine mammal hosts. Many species of marine fish are used by these larval anisakids as paratenic or intermediate hosts, and these are capable of infecting human consumers, causing public health, economic and social concerns worldwide. They are responsible for a fish-borne zoonosis called anisakidosis and associated allergic problems in humans, monetary loss to the fishing industry and the avoidance of fish products by consumers. An understanding of the parasite/fish relationship and the development of improved methods and tools for parasite detection are important to provide better insights to their biology, as well as technology to control the quality and safety of fishery products. In the present study, Anisakis simplex s.l. infection levels and location within the viscera of Atlantic herring (Clupea harengus) were visually investigated to provide a better understanding of settlement behaviour within the host fish, and to test if Magnetic Resonance Imaging (MRI) is a useful technology to detect anisakids in the viscera of whole herring. MRI potential to detect anisakids in fish muscle was also tested. Visual inspection determined an ascaridoid prevalence of 76% (>90% A. simplex s.l. (likely A. simplex s.s.) in herring viscera, with 69% of the parasites located outside the hind stomach and intestine areas. MRI demonstrated a capacity to detect A. simplex s.l. accumulations in the viscera of whole herring, A. simplex s.l. larvae and their movements within fish muscle, and Pseudoterranova sp. larva in fish muscle. Visual inspection of herring showed that A. simplex s.l. larvae frequently accumulate in the posterior end of the terminal blind sac of the stomach, around the ductus pneumaticus, as had previously been reported in herring and other clupeids. It has been hypothesized that the food digestion process, visceral organ topography and physicochemical conditions of fish tissues may play an important role in determining the encapsulation site and in vivo migration behaviours of A. simplex s.l. larvae within fish hosts. MRI showed potential to investigate the occurrence of anisakids and possibly other macroparasites of fish, and to detect anisakids in fishery products in situ, in a 3D environment and in a non-invasive and non-destructive way. Further investigation is required to determine if MRI can be used as a routine anisakid inspection tool by the fishing industry.     

Quantitative microbiological risk assessment (QMRA) of food-borne zoonoses at the European level

At EU level, the European Food Safety Authority (EFSA) is the body responsible for risk assessment in the field of food and feed safety. Following a request from European Union (EU) risk managers, the EFSA Panel on Biological Hazards (BIOHAZ) provided, for the first time, two EU-wide farm-to-fork quantitative microbiological risk assessments (QMRA), with regard to Salmonella in slaughter and breeder pigs and Campylobacter in broilers. The Scientific Opinion on a QMRA of Salmonella in pigs represented a major step forward in terms of modelling from farm to consumption as it took into account the variability between and within EU Member States. This QMRA model was developed to estimate the prevalence of infection/contamination and the microbial load from the farm to the point of consumption (exposure) and then estimating the probability of infection. It was used to investigate the effect of interventions to control Salmonella in pigs at different points of the food chain and resulted in a hierarchy of suggested on farm and slaughterhouse control measures, with estimates of the reduction of human cases. To model the effect of interventions from farm to fork on the incidence of human campylobacteriosis, a QMRA model was developed and used in the framework of the Scientific Opinion on a QMRA of Campylobacter in broiler meat. Reductions to the public health risk of campylobacteriosis could be achieved through a variety of interventions, both in primary production or at the slaughterhouse, with different impacts. Reductions of public health risk using targets at primary production or microbiological criteria were also estimated through modelling using additional models. QMRA of food-borne pathogens at European level has proven a useful/good tool to enable risk managers to evaluate the feasibility and the cost-benefit ratio of introducing control measures and targets to further protect public health of European consumers.     

Irradiation: a critical control point in ensuring the microbiological safety of raw foods

The widespread and increasing incidence of foodborne diseases and the resultant social and economic impact on the human population have brought food safety to the forefront of public health concerns. This has prompted public health authorities worldwide to reassess their methods of food safety assurance, and to resort to a more cost-effective, preventive method that is known as hazard analysis and critical control point (HACCP). Ensuring food safety depends on effective control measures, i.e., methods to prevent food contamination and, when necessary, decontamination. Present production methods cannot totally prevent food contamination, and the complexity of food handling and processing provides ample opportunity for contamination as well as survival and growth of pathogenic organisms. It is also unlikely that the methods of production can ensure foods totally free from in the near future, for many pathogens are part of the normal flora of the environment. The application of an HACCP-based approach as a method for the management of hazards of the food chain demonstrates the need for applying a cold decontamination treatment as a control measure in the production of foods which are to be marketed raw or minimally processed. Irradiation (increasingly referred to as “cold pasteurization”) is such a control measure in the production of several types of raw or minimally processed foods such as poultry, meat and meat products, fish, seafood, and fruits and vegetables. In the production of these foodstuffs, irradiation may thus be a critical control point (CCP). It has the potential to eliminate vegetative forms of bacterial pathogens as well as parasites. Moreover, irradiation fulfils other criteria for a CCP, i.e., critical limits (minimum and maximum doses) can be established and monitored, and process control is well known. Corrective actions can also be taken when necessary. Irradiation is a safe technology and has been recognized as such by the FAO/WHO Codex Alimentarius Commission. It certainly merits the attention of industry and public health authorities. Today, 40 countries permit the irradiation of one or more foodstuffs: 12 countries have approved its use for pathogen control in poultry, 8 other for use in meats, and 13 in fish and seafood.     

Tracing contamination of Methicillin-resistant Staphylococcus aureus (MRSA) into seafood marketing chain by staphylococcal protein A typing

Methicillin resistant Staphylococcus aureus (MRSA) is not a resident flora of fish. Its presence is endorsed to post-harvest contamination viz., handlers, handling equipment and environment. Earlier investigation revealed the presence of MRSA in seafood sold in retail fish markets in Kerala, India. Further studies were conducted to understand and identify the source of contamination into seafood sold in the retail markets by a pilot study. Seventeen samples which includes seafood and fishery environment samples from a landing centre and a retail fish market were collected to identify the source of contamination of MRSA. The whole experiment was repeated with same sampling plan for validation of the procedure, a week later from the same landing centre to the point of sale at fish market. MRSA was isolated from 35.2% to 23.5% of samples during first and second visits respectively. spa typing of the MRSA isolates revealed that MRSA from the landing centre (t311 and t15669) were carried to the retail fish market. Ice and water were the probable source for contamination during handling at the landing centre. This is first study to trace the source of contamination of MRSA in seafood and fishery environment. It is imperative that spa typing can be implemented for studying the local spread of MRSA clones at specific geographical locations only after establishing its diversity. To better understand the complexity of local spread of MRSA and reproducibility of this experiment, studies has to be conducted in other landing centre and retail markets.     

Qualitative risk assessment of introduction of anisakid larvae in Atlantic salmon (Salmo salar) farms and commercialization of products infected with viable nematodes

In this study, the likelihood of introduction of anisakid larvae in Atlantic salmon (Salmo salar) farms and subsequent commercialization of fish infected by at least one vital parasite was formally assessed using an adaptation of the risk assessment framework for importation of animals and animal products of the World Organization for Animal Health (OIE). The different plausible pathways were identified and outlined. The most recent information concerning the parasite, its definitive, intermediate hosts and the farming practices typical of Atlantic salmon farms were reviewed and used to assess the likelihoods of each key step along the pathways. A matrix for the conditional probabilities was adopted to combine the qualitative estimations and obtain an objective and transparent overall risk of introduction along each route. In order to avoid misinterpretation and overconfidence on the outcome, the uncertainties surrounding the estimations were considered. The only situation for which the assessed risk in a typical Atlantic salmon farm was deemed to be non-negligible involved the ingestion of infected hosts that penetrate the harvesting cages. In this event, the overall risk was estimated as ‘Very Low’ with a high degree of uncertainty because of the scarcity of information in some of the key steps along the pathway. However, the scientific evidence in support of the overall estimation suggests that the availability of additional data would be unlikely to increase the final estimated risk. On the basis of the available information of the system, the estimated risk of introduction and commercialization of farmed Atlantic salmons infected by viable nematodes resulted ‘Very Low’ even under a conservative approach. The proposed qualitative model is an objective and transparent method to assess the risk when data and information are scarce and provides a framework for the qualitative assessment of the introduction of alive parasites in aquaculture/mariculture implants and subsequent commercialization of infested fishery products. The framework could be easily adapted to other parasite-host interactions besides anisakid nematodes in Farmed Atlantic salmons.     

The international risk governance council framework and its application to Listeria monocytogenes in soft cheese made from unpasteurised milk

The International Risk Governance Council (IRGC) developed a Risk Governance Framework whose purpose is to help policy makers, regulators and risk managers both understand the concept of risk governance and apply it to their handling of risks. The Framework allows input from the scientific and public health perspective as well as the less-easily measurable side of freedom of choice. For food, the public wants many choices and less regulation, but at the same time safe products to consume. These ideals are at times in conflict. One example of this is the demand for soft cheese made from unpasteurised milk which may be contaminated with L. monocytogenes or other pathogens. Distinguishing between simple, complex, uncertain and ambiguous risks can help to design a risk management strategy. Public health advocates typically see management of soft cheese made from unpasteurised milk as a simple risk (protection of health), whereas for a small but vociferous proportion of the public sees it more of an uncertain or ambiguous one (choice trumps the slight risk of illness). The implications of five management options for the cheese are discussed. The new Codex Guidelines tend to put these cheeses into Option 1, a strict control on the presence of the pathogen, which will be ignored by the cheese aficionados. Other options are worth exploring to give some more choice but at low risk.     

Inorganic arsenic in rice and rice-based diets: Health risk assessment

Total and inorganic arsenic (As) content in rice and rice-based diets (n = 59) obtained from supermarkets in South Australia were studied to investigate the contamination levels and whether consumption of these products pose potential health risks to young children and adults. Results show that of the 59 rice-based products, 31 (53%) exceeded the EU recommended value (100 μg/kg) of As for young children and 13 (22%) samples had higher than maximum level of 200 μg/kg recommended for adults. Arsenic content varies as rice crackers > baby rice > rice cakes > puffed rice > other rice-based snacks > ready-to-eat rice. Of the 6 categories of rice-based products, except ready-to-eat rice, all others exceeded the EU recommended value for young children. Even manufacture recommended servings deliver significant amounts (0.56–6.87 μg) of inorganic As. These amounts are within the range of BMDL₀₁ values indicated by the European Food Safety Authority (EFSA), which means the risk cannot be avoided for young children and adults considering the levels of total and inorganic As in rice-based products.     

Species identification in anchovy pastes from the market by PCR-RFLP technique

The wide variety of marketed fishery products sometimes makes the recognition of species difficult, especially in processed food. So the consumer can be exposed to several commercial and health frauds. The European law obliges to indicate the species by specific commercial names only in fresh and prepared fishery products, but not in processed ones, in which the species can be named using generic denominations. In the present study, the use of a PCR-RFLP technique for species identification in anchovy paste, a widespread product in Italy, is described. After the PCR amplification of a common 272 bp fragment of mitochondrial cytochrome b gene from different fish species used as standards (European anchovy, European pilchard, European sprat, twaite shad, round sardinella, Atlantic horse mackerel and Mediterranean horse mackerel), the amplicons were digested using four restriction enzymes (MspI, HincII, Eam1104I and Alw26I). These patterns were compared with patterns from DNA extracted from anchovy pastes purchased from the market enabling to detect the presence of species different from anchovy (Engraulis encrasicolus) in some of them. The suitability of extending the obligation to indicate the species using specific commercial denominations even to processed fishery products is also discussed.     

Prevention and control of hazards in seafood

Seafood is high on the list of foods transmitting disease. However, the food safety issues are highly focussed and more than 80% of all seafood-borne outbreaks are related to biotoxins (ciguatoxin), scombrotoxin or the consumption of raw molluscan shellfish. The safety hazards in seafood production are listed and discussed. It is pointed out that there are serious safety concerns related to the consumption of raw fish and shellfish due to the presence of biological (bacteria, virus, parasites) and chemical (biotoxins) hazards. These hazards are present in the fish and shellfish pre-harvest and are difficult or impossible to control by applying presently available preventive measures. In contrast, the hazards related to contamination, recontamination or survival of biological hazards during processing are well-defined and can be controlled by applying Good Manufacturing Practice (GMP), Good Hygiene Practice (GHP) and a well designed HACCP-programme. Similarly, the means to prevent the growth of pathogenic microorganisms during distribution and storage of the final products are – with a few exceptions – available. Proper application of well-known preservative parameters including temperature is able to control growth of most pathogens. When this is not the always case, for example inhibition of Listeria monocytogenes in lightly preserved fish products, it is recommended to limit the stated shelf-life of these products to a period of no-growth for the pathogen of concern. There is a good agreement between the trends shown in disease statistics, the hazard analysis and the qualitative risk assessment of the various fish products. It is recommended that consumers should be informed of the risk of eating raw seafood – particularly molluscan shellfish and certain freshwater fish.     

The SAFE FOODS Risk Analysis Framework suitable for GMOs? A case study

This paper describes the current EU regulatory framework for risk analysis of genetically modified (GM) crop cultivation and market introduction of derived food/feed. Furthermore the risk assessment strategies for GM crops and derived food/feed as designed by the European Food Safety Authority (EFSA) are described on which international agreement exists. Existing flaws in the EU regulatory framework for GMOs have been identified and proposals are put forward to improve current risk analysis procedures for GMOs by taking the SAFE FOODS Risk Analysis Framework into account. The SAFE FOODS framework describes an iterative decision-making process with four distinct stages i.e. framing, risk–benefit assessment, evaluation, and risk management which includes decision-making, and implementation, and a final review stage. Three major changes compared to current risk analyses practices are proposed, i.e. (i) the addition of a formal framing stage, during which problem formulation and the objectives of the risk analysis are established, (ii) enlargement of the scope of the risk assessment, by including the assessment of potential benefits, and an impact analysis of social and economic aspects, and (iii) addition of a formal evaluation stage, in order to weigh risks, costs and benefits and their distribution. Furthermore a broader participation of certain entities, organisations and individual citizens in specific segments of the risk analysis process, in particular in the framing and evaluation stage, is proposed. The proposed changes in current risk analyses practises may contribute to restore consumer confidence in risk analysis process of GMOs in the EU.