Risk Assessment of AFM1 in Raw Milk and Dairy Products Produced in Armenia, a Caucasus Region Country: A Pilot Study.

This paper presents the first assessment of dietary exposure to aflatoxin M1 (AFM1) and associated health risks through milk and dairy product consumption in Armenia. Data on AFM1 in raw milk were obtained from an annual residue monitoring program. Additionally, commonly consumed dairy products (pasteurized milk, cheese, sour cream, curd cheese) were sampled, considering the sources of raw milk used by dairy companies. Per capita consumption of raw milk was sourced from national food balance databases, while individual consumption data for dairy products was collected via a 24 h recall survey with 1400 adult respondents. Detectable levels of AFM1 were observed in 7.14% of raw milk samples (up to 0.334 µg/kg) and, albeit at lower amounts (up to 0.009 µg/kg), in 30% and 40% of sour cream and curd cheese, respectively. The AFM1 levels were lower than the national maximum permitted level (0.5 µg/kg); however, levels in raw milk exceeded the EU ML (0.05 µg/kg). The estimated margin of exposure values for dairy products indicated no significant risk, whereas a reasonable worst-case estimate, using the measurable levels of AFM1 in raw milk consumption indicated a potential public health concern. This study provides a scientific basis for evaluating aflatoxin issues in the Caucasus area.

Guidance on the assessment of the safety of feed additives for the users.

This guidance document is intended to assist the applicant in the preparation and the presentation of an application, as foreseen in Article 7.6 of Regulation (EC) No 1831/2003, for the authorisation of additives for use in animal nutrition. It specifically covers the assessment of the safety for the users.

Exposure Assessment of Nitrofuran Metabolites in Fish and Honey Produced in Armenia: A Pilot Investigation.

In Armenia, the presence of nitrofuran residues in food products is unacceptable for both domestic sales and export. However, food may contain nitrofuran metabolites (NMs) due to the illegal use of these drugs in the agrofarming practice. This study aimed to identify NMs as the marker residues for nitrofurans in fish and honey produced in Armenia and assess the potential health risks associated with consuming these foods. The commodities studied were natural honey and three species of farmed fish produced by various regions nationwide. Concentrations of the marker metabolites (3-amino-2-oxazolidinone (AOZ), 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ), 1-aminohydantoin (AHD), and semicarbazide (SEM)) were determined through an enzyme-linked immunosorbent assay (ELISA) and verified using liquid chromatography-mass spectrometry (LC-MS/MS). Consumer groups were identified based on their average daily intake of foods. Health risk was assessed by calculating the margin of exposure (MOE). Reference values for health risk assessment were obtained from the European Food Safety Authority (EFSA). Results showed that 33.3% of fish samples and 44.4% of honey samples contained NMs, the mean concentrations ranging from 0.05 µg/kg to 0.52 µg/kg. All MOE values obtained were over 10,000, indicating that the detected concentrations of NMs in fish and honey produced in Armenia pose no health risk to consumers. However, these results highlight the illicit use of highly toxic substances and the need for improved control of farming practices.

Assessment of Heme and Non-Heme Iron Intake and Its Dietary Sources among Adults in Armenia.

Adequate dietary iron (Fe) intake is crucial for preventing Fe-deficient anemia, a recognized global public health concern which is important in Armenia. This study aimed to analyze the intake of Fe, both heme (from animal tissues) and non-heme (more prevalent, but less efficiently absorbed), as well as the Fe dietary sources, among adults in a representative national sample in Armenia. The study was conducted on 1400 individuals aged 18-80 and above, who were enrolled from all regions of Armenia. The Fe intake was assessed through a 24 h dietary recall survey, while Fe occurrence was determined through atomic absorption spectrophotometry (AAS). The results showed a high proportion of adults with a Fe intake lower than the average requirements set by EFSA (65%, 80% and 85% of males, total females and females at fertile age, respectively). Main Fe sources were bread, fruits and vegetables; heme Fe accounted only for <5% of total Fe intake. Compared to males, females had a lower intake of all forms of Fe (p < 0.05). Significant differences were observed in the intake of different forms of Fe between regions (p < 0.05), while the age-group 36-55 years had higher intakes of total Fe. Our data call for comprehensive nutritional security strategies in order to reduce iron deficiency in Armenia, that represents a public health concern.

Risk assessment of dietary exposure to trace elements that are reproductive toxicants: Lead, molybdenum and nickel. The case study of Armenia.

The research aimed to assess dietary exposure to developmental toxicants (Mo, Ni, Pb) among the Armenian adult female population of reproductive age (18-49 years). Commonly consumed foods with a daily intake of more than 1 g in Armenia have been selected to assess the occurrence of Mo, Ni, and Pb. Food consumption data among the adult population in Armenia were collected in the frame of the national survey via 24-h recall method. Estimated daily intakes (EDI) and associated potential health risks for both mean and high (95 percentile) consumers were assessed based on the health-based guidance values (HBGVs). None of the EDI values for the developmental toxicants via individual food consumption exceeded the HBGVs, however, the EDI of Pb in the case of aggregate consumption of all food products exceeded HBGV of 0.5 µg/kg b.w./day, indicating possible concerns for the neurodevelopmental effects. Noticeably, the intake of Pb through some individual food items (cheese curd, beef and veal, pelmeni and khinkali, black coffee, tap water) and the aggregate consumption of all the studied foods led to a Margin of Exposure lower than 10 compared to HBGV.This study is the first one carried out on dietary exposure to developmental toxicants among women of fertile age in a Caucasus country. The outcomes prompt the need to investigate sources of Pb pollution in foods consumed in Armenia (natural or human-derived environmental pollution, as well as food contact materials, etc.) and may pave the way for similar studies in the Caucasus region.

Risk assessment of potentially toxic trace elements via consumption of dairy products sold in the city of Yerevan, Armenia.

The study aimed to investigate the potentially toxic trace elements (PTEs) Pb, Cd, Hg, Mo, and Cu in dairy products sold in Yerevan's markets and to assess the related exposure and possible health risks among the adult population residing in the capital city of Armenia, Yerevan. The PTEs contents were determined in widely consumed dairy products (pasteurized milk, sour cream, matsoun and cheese from cow milk) using graphite furnace atomic absorption spectrometry. PTE contents were in the range of 7.90E-04 (Pb in pasteurized milk) to 6.75E-01 (Cu in cheese) mg/kg. Food consumption was assessed by the 24-h dietary recall method. The PTE daily intakes and margin of exposure (MOE) were calculated by the deterministic approach. The results indicated potential health concerns for the contribution of dairy products to the overall dietary intake of Pb and Cd. MOEs of Pb and Cd (8.71 and 8.80, respectively) estimated for high consumers of cheese (90 g/day and more) were lower than 10. The pilot data set provides the first comprehensive appraisal on the dairy products contamination by PTEs in the Caucasus region. The results point out further attention to sources of Pb and Cd in Armenian dairy products, including environment and farm characteristics.

Risk-Benefit Assessment of Feed Additives in the One Health Perspective.

Safety and sustainability of animal feeds is a pillar of the safety of the entire food chain. Feed additive assessment incorporates consumer safety as well as animal health and welfare, which, in turn, can affect productivity and hence food security. The safety of feed users and the environment are other important components of the assessment process which, therefore, builds on a One Health perspective. In several instances the assessment entails a balanced assessment of benefits and risks for humans, animals and/or the environment. Three case studies are selected to discuss issues for a consistent framework on Risk-Benefit Assessment (RBA) of feed additives, based on EFSA opinions and literature: (a) Supplementation of feeds with trace elements with recognized human toxicity (cobalt, iodine) - RBA question: can use levels, hence human exposure, be reduced without increasing the risk of deficiency in animals?; (b) Aflatoxin binders in dairy animals - RBA question: can the use reduce the risk for human health due to aflatoxin M1, without unexpected adverse effects for animals or humans?; (c) Use of formaldehyde as preservative in feedstuffs to prevent microbial contamination - RBA question: is the reduction of microbiological risks outweighed by risks for the consumers, farmed animals or the workers? The case studies indicate that the safety of use of feed additives can involve RBA considerations which fit into a One Health perspective. As in other RBA circumstances, the main issues are defining the question and finding "metrics" that allow a R/B comparison; in the case of feed additives, R and B may concern different species (farm animals and humans). A robust assessment of animal requirements, together with sustainability considerations, might be a significant driving force for a RBA leading to a safe and effective use.

Human health risk-benefit assessment of fish and other seafood: a scoping review.

Fish and other seafood are important sources of nutrients, but they are also sources of chemical contaminants that may cause adverse health effects. This article aimed to identify existing risk-benefit assessments (RBA) of fish, shellfish, and other seafood, compare methodologies, discuss differences and commonalities in findings, and identify limitations and ways forward for future studies. We conducted a scoping review of the scientific literature of studies in all languages published from 2000 through April 2019. We identified 106 RBA of fish and other seafood across Europe, Asia, North America, Africa, and at the global level. Studies were heterogeneous in terms of types of fish and other seafood considered, beneficial and adverse compounds assessed, and overall methodology. Collected data showed that a diet consisting of a variety of lean and fatty fish and other seafood is recommended for the overall population and that women of childbearing age and children should limit the consumption of fish and other seafood types that have a high likelihood of contamination. Our review emphasizes the need for evidence-based, up-to-date, and harmonized approaches in RBA in general.

Potential harm caused by physicians' a-priori beliefs in the clinical effectiveness of hydroxychloroquine and its impact on clinical and economic outcome - A simulation approach.

BACKGROUND: Despite growing controversies around Hydroxychloroquine's effectiveness, the drug is still widely prescribed by clinicians to treat COVID19 patients. Therapeutic judgment under uncertainty and imperfect information may be influenced by personal preference, whereby individuals, to confirm a-priori beliefs, may propose drugs without knowing the clinical benefit. To estimate this disconnect between available evidence and prescribing behavior, we created a Bayesian model analyzing a-priori optimistic belief of physicians in Hydroxychloroquine's effectiveness. METHODOLOGY: We created a Bayesian model to simulate the impact of different a-priori beliefs related to Hydroxychloroquine's effectiveness on clinical and economic outcome. RESULTS: Our hypothetical results indicate no significant difference in treatment effect (combined survival benefit and harm) up to a presumed drug's effectiveness level of 20%, with younger individuals being negatively affected by the treatment (RR 0.82, 0.55-1.2; (0.95 (1.1) % expected adverse events versus 0.05 (0.98) % expected death prevented). Simulated cost data indicate overall hospital cost (medicine, hospital stay, complication) of 18.361,41€ per hospitalized patient receiving Hydroxychloroquine treatment. CONCLUSION: Off-label use of Hydroxychloroquine needs a rational, objective and datadriven evaluation, as personal preferences may be flawed and cause harm to patients and to society.

25th anniversary of the Berlin workshop on developmental toxicology: DevTox database update, challenges in risk assessment of developmental neurotoxicity and alternative methodologies in bone development and growth.

25 years after the first Berlin Workshop on Developmental Toxicity this 10th Berlin Workshop aimed to bring together international experts from authorities, academia and industry to consider scientific, methodologic and regulatory aspects in risk assessment of developmental toxicity and to debate alternative strategies in testing developmental effects in the future. Proposals for improvement of the categorization of developmental effects were discussed as well as the update of the DevTox database as valuable tool for harmonization. The development of adverse outcome pathways relevant to developmental neurotoxicity (DNT) was debated as a fundamental improvement to guide the screening and testing for DNT using alternatives to animal methods. A further focus was the implementation of an in vitro mechanism-based battery, which can support various regulatory applications associated with the assessment of chemicals and mixtures. More interdisciplinary and translation research should be initiated to accelerate the development of new technologies to test developmental toxicity. Technologies in the pipeline are (i) high throughput imaging techniques, (ii) models for DNT screening tests, (iii) use of computer tomography for assessment of thoracolumbar supernumerary ribs in animal models, and (iv) 3D biofabrication of bone development and regeneration tissue models. In addition, increased collaboration with the medical community was suggested to improve the relevance of test results to humans and identify more clinically relevant endpoints. Finally, the participants agreed that this conference facilitated better understanding innovative approaches that can be useful for the identification of developmental health risks due to exposure to chemical substances.

Risk assessment of dietary exposure to potentially toxic trace elements in emerging countries: A pilot study on intake via flour-based products in Yerevan, Armenia.

Flour-based products, one staple food items of Armenians' diet, can be major contributors to the total exposure to trace elements. Therefore, this pilot study estimated the exposure to potentially toxic trace elements (Pb, Cd, Hg, Ni, Mo, Cu), and the related health risks, through consumption of the main flour-based products (white bread, lavash, pasta, cookies) in the capital city Yerevan. The TE contents in market samples were analyzed using atomic absorption spectrometry. The product consumption data were obtained via a 24-h recall survey amongst Yerevan adults (1272 respondents). Following the K-mean clustering test, three clusters were determined for each type of the product intake. The daily intakes and margin of exposure of trace elements were calculated by the deterministic approach. Considering the health-based guidance values and the obtained daily intakes, concerns for consumer safety were identified in case of dietary exposure to Ni. Moreover, the estimated of all studied elements highlighted that a consumer health concern (identified by margin of exposure less than 10) cannot be excluded in the case of white bread (mainly) as well as lavash and cookies consumption. This pilot study provides important highlights towards future total diet studies in Armenia.

Assessment of the application for renewal of authorisation of l-histidine monohydrochloride monohydrate produced with Escherichia coli NITE SD 00268 for salmonids and its extension of use to other fin fish.

Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on l-histidine monohydrochloride (HCl) monohydrate produced by fermentation using Escherichia coli NITE SD 00268 in the context of the renewal of the authorisation for salmonids when used as a nutritional additive. In addition, the applicant requested the extension of use of the additive for other fin fish. The applicant has provided evidence that the composition of the additive currently in the market complies with the conditions of authorisation. The production strain has been modified by conventional mutagenesis and it does not raise safety concerns. The use of l-histidine HCl monohydrate produced by fermentation using E. coli NITE SD 00268 is safe for salmonids and other fin fish when used as a nutritional additive to supplement the diet in appropriate amounts to cover the nutritional requirements, depending on the species, the physiological state of the animal, the performance level, the environmental conditions, the background amino acid composition of the unsupplemented diet and the status of some essential trace elements such as copper and zinc. The FEEDAP Panel considers the maximum total concentration of 1.7% histidine in feed for salmonids proposed by the applicant as safe. For other fin fish species, the level of 1.7% appears to cause adverse effects. Therefore, it is not possible to define a maximum concentration of histidine in fish other than salmonids as it depends on histidine nutritional requirements in the different fish species. The use of the authorised additive in salmonids production does not pose a risk for consumers, and the proposed maximum total concentration of 1.7% histidine in feed is considered safe for the consumer. l-Histidine HCl monohydrate produced using E. coli NITE SD 00268 supplemented at levels appropriate to cover the nutritional requirements of fish other than salmonids is considered safe for the consumer. The additive under assessment is not a skin irritant. In the absence of data, it is not possible to conclude on the potential of the additive to be toxic by inhalation, irritant to eyes or a skin sensitiser. The amino acid l-histidine is a natural component of plants and animals. The use of the additive under assessment in animal nutrition does not represent a risk to the environment. The additive is considered an efficacious source of the amino acid l-histidine for fish species.

Health risk assessment of toxicologically relevant residues in emerging countries: A pilot study on Malachite Green residues in farmed freshwater fish of Armenia.

Malachite Green (MG) has a worldwide application in aquaculture as a therapeutic agent; however, its use in food producing animals is illegal, due to potential carcinogenicity and persistence of residues. This pilot study, the first conducted in Armenia, aimed to determine the concentration of MG residues in flesh of fish grown in artificial ponds of Armenia and conduct dietary exposure assessment to characterize possible health risks to consumers. Detection of MG residues, including the major metabolite leucomalachite, was carried out in 29 fish composite samples by ELISA. The results were confirmed by LC-MS/MS. To determine fish consumption values, a food frequency questionnaire was used. Possible health risks were evaluated by calculating the Margin of Exposure (MOE) based on BMDL of 13 (neoplastic effects) and 6 (non-neoplastic effects) mg/kg bw. In 34.5% of the investigated fish samples MG residues exceeded the minimum required performance limit. For BMDL10 and BMDL0.5, the MOEs ranged 3.36E+06-3.37E+07 and 1.55E+06-1.55E+07, respectively. The MOE for neoplastic effects was more than 10,000 and for non-neoplastic effects was more than 100. The results do not indicate public health concerns. However, the results highlight issues concerning the illegal use of MG in Armenian aquaculture, which deserves further attention.

Update of the DevTox data database for harmonized risk assessment and alternative methodologies in developmental toxicology: Report of the 9th Berlin Workshop on Developmental Toxicity.

Representatives of applied science (e.g. governmental organizations, academia, and industry) met to discuss the progress towards a harmonized human health risk assessment in developmental toxicology of plant protection products, biocidal products, and other environmental chemicals at the 9th Berlin Workshop on Developmental Toxicity held in September 2018. Within the focus of the scientific discussion were the future of in-vitro methods for developmental and reproductive toxicology, the potential relevance of alternative species in testing of developmental effects, and risk and hazard assessment of developmental and endocrine effects. Furthermore, the need for a harmonized terminology for classification of anomalies in laboratory animals in developmental toxicity studies aiming for human health risk assessment was determined. Here, the DevTox database was identified as an extremely valuable tool. Overall, the participants agreed that still one of the biggest challenges for testing developmental toxicity in the 21st century is the development of animal-free test strategies and alternatives to animal testing that could provide human-relevant information in a rapid, efficient, and mechanistically informative manner.

Assessment of the application for renewal of authorisation of selenomethionine produced by Saccharomyces cerevisiae NCYC R397 for all animal species.

The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the application for renewal of authorisation of organic form of selenium produced by Saccharomyces cerevisiae NCYC R397 (Alkosel®) for all animal species. The FEEDAP Panel has delivered two opinions (on 2007 and 2016) on the safety and efficacy of the additive. The additive is characterised as organic selenium mainly selenomethionine (63%); it was initially authorised in 2007 with a content of 2,000-2,400 mg Se/kg (97-99 % of organic selenium) and in 2017 the authorisation was further amended to introduce a selenium range of 2,000-3,500 mg Se/kg. Additionally, in 2013, the initial authorising Regulation was modified to introduce a maximum supplementation selenium from this additive in feed (0.2 mg Se from Alkosel®/kg feed). The evidence provided indicates that the additive currently in the market complies with the conditions of authorisation. No new evidence was found that would make the FEEDAP Panel reconsidering its previous conclusions in the safety for target species, consumers and environment. In particular, the Panel confirms that the use of Alkosel® in animal nutrition does not pose a risk to consumers provided that the maximum selenium supplementation of 0.2 mg/kg feed from Alkosel® is not exceeded, yet respecting the maximum total selenium in feed of 0.5 mg/kg. Data on the characterisation of the additive and studies on skin/eyes effects led the Panel reconsider the safety for the user, concluding that Alkosel® is (i) hazardous upon inhalation, (ii) non-irritant to skin and considered irritant for the eyes and mucosae, and (iii) a dermal sensitiser and likely a respiratory sensitiser. A recommendation regarding the denomination of the additive under assessment was proposed by the Panel.

Assessment of the application for renewal of authorisation of Lantharenol® (lanthanum carbonate octahydrate) for cats.

Lantharenol® is a feed additive consisting of lanthanum carbonate octahydrate. It is currently authorised as a zootechnical additive (decrease in phosphorous excretion via urine) for cats; this opinion concerns the renewal of the authorisation. In 2007, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) adopted an opinion on the safety and efficacy of Lantharenol® as a feed additive for cats. The applicant has provided data demonstrating that the additive complies with the conditions of authorisation. The available evidence, including the results of the post-market monitoring plan, confirms that the use of Lantharenol® in cat food at the maximum authorised concentration of 7.5 g/kg of complete feed is safe for cats. In the absence of new data, the FEEDAP Panel reiterates its previous conclusion that Lantharenol® does not pose a risk by inhalation and is not an irritant to skin or eye and not a dermal sensitiser. The FEEDAP Panel considers that there is no need for a continuation of a specific post-market monitoring plan.

Assessment of the application for renewal of authorisation of l-arginine produced by fermentation using Corynebacterium glutamicum NITE SD 00285 for all animal species.

l-Arginine is an amino acid that is conditionally essential for mammalian neonates, some strict carnivores, birds and fish. The subject of this opinion is a request for renewal of authorisation of l-arginine produced by a strain of Corynebacterium glutamicum. The strain designation has changed to its new deposition number, NITE SD 00285, but the strain is otherwise unchanged from the previous opinion. It is not genetically modified and possesses no antibiotic resistance of safety concern. Minor changes in downstream processing following fermentation have been made. The FEEDAP Panel notes that two out of five batches did not meet the specification of the current authorisation (minimum 98% on a dry matter basis). The FEEDAP Panel concludes that l-arginine produced by fermentation to C. glutamicum NITE SD 00285 remains safe for the target species, consumers of products from animals fed the additive and the environment under the approved conditions of authorisation. The additive is considered as irritant to skin, corrosive to eyes and poses a risk by inhalation.

Vaccines: An achievement of civilization, a human right, our health insurance for the future.

Vaccines have made a key, cost-effective contribution to the prolongation of life expectancy and quality. Here we summarize challenges facing vaccinology and immunology at the level of society, scientific innovation, and technology in a global health perspective. We argue that vaccines represent a safety belt and life insurance for humankind.