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.

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.

Health risks of heavy metals in food and their economic burden in Armenia.

Mining leads to excessive heavy metal contamination of agricultural products in Armenia and can adversely impact human health. We quantify the concentrations of toxic heavy metals (lead, nickel, cadmium, and mercury) in food sampled from local markets of the capital city, Yerevan. We combine these measured concentrations with data from a diet survey of 1,195 people. The results provide an estimate of people's heavy metal intake in a typical day. The study finds that only dietary exposure to lead raises consumer safety concerns. More than 50 % of the population have daily intakes of lead that exceed the benchmark dose level (BMDL10) of 0.63 µg/kg b.w./day established by European Food Safety Authority. Moreover, risk assessment results indicate that approximately 96 % of the population has dietary exposure values for lead that exceed the threshold level, with animal products as the primary source. The lead exposure observed in sampled population has the potential to increase blood pressure on average by 0.54 mmHg, lower IQ levels by 1.2 point, and reduce lifetime earnings by $5000. The study reveals a strong need to determine the specific pathways by which lead enters the food supply, to mitigate the excess exposures, and to reduce the potential or severity of the resulting adverse health impacts.

Risk Characterization of the Armenian Population to Nickel: Application of Deterministic and Probabilistic Approaches to a Total Diet Study in Yerevan City.

Nickel (Ni) is a widespread metal that occurs in food and drinking water from both natural and anthropogenic sources. Oral exposure to Ni can induce a variety of adverse effects; the European Food Safety Authority established a tolerable daily intake (TDI) of 13 µg/kg bw and a lowest-observed-adverse-effect level (LOAEL) of 4.3 µg/kg bw to assess the risk of allergic reactions upon acute exposure. This study, the first conducted in Armenia, aimed to assess the dietary exposure of the adult Yerevan population (1272 subjects of both sexes) to Ni in a total diet study (TDS). Detection of Ni was carried out using atomic absorption spectrometry. To determine food consumption values, a 24-h recall survey was used. Following the K-means clustering test, two clusters were determined for food product intake. For the risk characterization of acute oral exposure, the margin of exposure (MOE) was calculated using both deterministic and probabilistic (Monte Carlo method) approaches. The average total exposure was 4.396 µg/kg bw, with limited influence by age and gender. The main contributors were "fruits and vegetables" followed by "bread and flour-based products": the total intake would be 5.11 µg/kg bw for a woman with high consumption of fruits and vegetables. Hence, the estimated chronic dietary exposure was below the TDI, irrespective of age and gender groups, and including high consumers. However, acute oral exposure estimates led to MOE values of less than 30 for most food products, indicating potential health concerns for Ni-sensitized individuals. The Monte Carlo approach indicated that the probability of occurrence of MOE lower than 30 was very high in the case of beef/veal, pork and chicken meat, eggs, and fish, alongside vegetable sources such as buckwheat, tomato, watermelon/melon, and potatoes. The findings prompt an investigation of Ni sources in the target foods in the Caucasus area.

Assessing Dietary Intakes from Household Budget Survey in Armenia, 2008-2019.

Household budget surveys are used regularly to estimate dietary intakes. The study aims to assess the trends in food consumption and nutrient intake, according to 14 dietary indicators from household budget surveys in Armenia. Data on food consumption was obtained from Armenian Integrated Living Conditions Surveys, 2008-2019. The results indicate that the consumption of all types of foods, including plant-origin has decreased, whereas the consumption of foods of animal origin has mostly stayed stable. Over time, the energy and macronutrient intakes of Armenians have decreased, while the contribution of each food group to total energy and nutrient intake has not changed. More than 50% of total energy, protein, and carbohydrate intake is attributable to cereals and bakery products. The population is characterized by macronutrient variations; the amounts of energy and carbohydrate intake are below the recommended values set by WHO/FAO, total fat intake is at the highest recommended level, while the amount of protein exceeds the threshold. Based on the findings there is an urgent need to increase awareness of nutritional requirements and a need to change widespread dietary practices, such as irregular meal intake and omission of breakfast.

Risk assessment of uptake of trace elements through consumption of cereals: a pilot study in Yerevan, Armenia.

The study aimed to assess the dietary exposure and related human health risks associated with trace elements through the intake of staple cereals, including buckwheat, rice, and emmer. The contents of Lead (Pb), Cadmium (Cd), Mercury (Hg), Nickel (Ni), Molybdenum (Mo), Iron (Fe), and Copper (Cu) were determined using Atomic Absorption Spectrometry. Cereal consumption data were obtained through a Semi-Quantitative Food Frequency Questionnaire amongst the Yerevan adult population. Concentrations of Pb, Cd, Hg, Ni, Mo, Fe and Cu were 0.75-5.56 µg/kg, 1.21-6 µg/kg, 6.7-6.99 µg/kg, 50.6-111 µg/kg, 20-429 µg/kg, 2770-30500 µg/kg and 601-1720 µg/kg respectively. The estimated daily intakes (EDI) of all studied trace elements did not exceed the health-based guidance values. The margin of exposure (MOE) values of Pb, Cd, Hg, and Mo for all clusters were above the threshold (>10 for Pb, Cd, and >100 for Hg, Mo) and therefore did not indicate a health concern. Meanwhile, in the case of Ni exposure, the MOE values for the second and third clusters of buckwheat consumption and for the third cluster of emmer consumption were below the threshold (<10) indicating that a human health risk cannot be excluded. Future work is proposed.

Compositional features of Pb in agricultural soils and geochemical associations conditioning Pb contents in plants.

Soil geochemical data is compositional. Hence the studies targeting the potential of accumulation of toxic elements (TE) in plants have to consider the compositional nature of soil chemical environment. In this study, the combined application of compositional data analysis and geospatial mapping was used to investigate Pb geochemical associations in agricultural soils, revealing the link between these associations and Pb contents in plants, as well as identifying source-specific transfer of Pb from soil to plants. The obtained results showed that soil chemical composition was conditioned by the geological peculiarities of the study area and the potential sources of chemical elements' release. Particularly, k-means clustering and CoDa-biplot allows to identify three distinct subsamples and the application of HCA showed that both Pb soil and plants contents were in the same cluster in all subsamples. However, the geochemical association of elements in subsamples I and III suggested that Pb contents in plants were conditioned by the geochemical behaviors of carbonates whereas in subsample II Pb plant contents were presented in a geochemical association (K, Rb, Pb, and Zn) typical for both fertilizers and the potassium feldspar. The transfer factor (TF) for the comparatively higher values is observed for the subsample linked to K, Rb, Pb, and Zn geochemical association. At the same time, the negative influence of carbonates on the Pb availability in the plants was evidenced. The results of this study can serve as a good example for other investigations targeting the role of soil chemical elements compositional features in elements transfer to plant.

Trans-Fatty Acids in Fast-Food and Intake Assessment for Yerevan's Population, Armenia.

There are stringent regulations applicable for trans-fatty acid (TFA) limitations from food supply across the world. However, in Armenia, there is a scarcity of data on TFA content in food products and their consumption levels. Considering that fast-food is among the major contributors to TFA intake, this study aims to assess the dietary exposure of TFAs through the consumption of fast-food in Yerevan, Armenia. Eleven types of fast-food were included in the study. The Food Frequency Questionnaire (FFQ) was used to evaluate daily fast-food consumption. TFA contents in samples were determined using gas chromatography-mass spectrometry. Mean daily fast-food consumption values ranged from 14.68 g/day to 76.09 g/day, with popcorn as the lowest and pastry as the highest consumed food. The study results indicate that the aggregate average daily intake (DI) of TFA is 0.303 g/day. Even though TFA DI values do not exceed the WHO limit of 1%, they substantially contribute to daily TFA intake and may exceed the limit when combined with other foods. Hence, it is recommended to carry out continuous monitoring of TFA content in the food supply to ensure consumer health protection.

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.

Toxic element contents and associated multi-medium health risk assessment in an area under continuous agricultural use.

Soils and contaminated plants are in the group of significant environmental pathways of human exposure to toxic elements (TEs). This study aimed to assess the soil-to-plant transfer of TEs (Pb, As, Cd, Hg), as well as the plausible health risks via different exposure pathways in the Armavir region of Armenia. The contents of TEs were determined in soil, fruit and vegetable samples using X-ray fluorescence and atomic absorption spectroscopy, respectively, and the soil-to-plant transfer of TEs and induced chronic non-carcinogenic risks were evaluated. The detected TE contents did not exceed the available national and international regulatory levels. Moreover, the evaluated soil-to-plant transfer of these TEs was negligible. The obtained results indicated the absence of a single-element non-carcinogenic risk via one single ingestion pathway. Meanwhile, in the case of multi-food and multi-soil ingestion, the combined non-carcinogenic risk estimates for Pb and As exceeded the precautionary level of 0.1 indicating a low health risk. Similarly, the multi-element hazard index (HI) showed a low level of non-carcinogenic risk through a multi-pathway ingestion. Thus, the outcomes highlighted that there was a need for precautionary measures to prevent plausible health issues for the adult population in Armavir.

The Effect of Trans Fatty Acids on Human Health: Regulation and Consumption Patterns.

Health effects of trans fatty acids (TFAs) on human organisms can vary according to their type, structure, composition, and origin. Even though the adverse health effects of industrial TFAs (iTFAs) have been widely discussed, the health effects of natural TFAs (nTFAs) are still questionable. Hence, it is important to review the literature and provide an overall picture on the health effects of different TFAs coming from industrial and ruminant sources, underlining those types that have adverse health effects as well as suggesting methods for reducing their harmful effects. Multiple databases (PubMed, Medline, Cochrane Library, etc.) were searched with the key words "trans fatty acid sources", "ruminant", "industrial", "conjugated trans linoleic acid", "human", "coronary heart disease", "cancer", etc. Reference lists of the studies were scanned discussing the health effects of iTFAs and nTFAs. The review of the literature showed that iTFAs are found to be more harmful than ruminant-produced nTFAs. Although several beneficial effects (such as reduced risk of diabetes) for nTFAs have been observed, they should be used with caution. Since during labeling it is usually not mentioned whether the TFAs contained in food are of industrial or natural origin, the general suggestion is to reduce their consumption.

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.

Carcinogenic and non-carcinogenic risk assessment of trace elements and POPs in honey from Shirak and Syunik regions of Armenia.

Honey is a highly nutritious natural product widely produced and consumed by people in Shirak and Syunik regions of Armenia. Unlike Shirak, Syunik is under the impact of mining industry. Since the environmental pollution can adversely impact the safety of honey and entail a probable risk to human health, it is important to evaluate the presence of potentially toxic trace elements in honey samples from both regions and draw comparisons. This study assesses the dietary exposure to trace elements and persistent organic pollutants through the intake of honey for the first time among people in Shirak and Syunik regions. 24-hour dietary recall method was used to investigate the consumption of honey. The presence of seven trace elements (Pb, Cd, As, Hg, Cu, Zn, and Ni) and persistent organic pollutants (hexachlorocyclohexane (HCH), dichlorodiphenyltrichloroethane (DDT) and its metabolites) were determined in honey samples using atomic absorption spectrometry and gas chromatography-mass spectrometry, respectively. In several samples, the concentrations of Cu were above the maximum allowable level. Non-carcinogenic risk values did not exceed the acceptable level, while carcinogenic risk values for Ni and As exceeded the risk level of 10-6 in both regions. Moreover, among the persistent organic pollutants, only the concentration of DDT in honey from Shirak was above the European Union maximum residue level.

Health Risk Assessment of Potentially Toxic Trace and Elements in Vegetables Grown Under the Impact of Kajaran Mining Complex.

Mining industry is one of the priority sectors of Armenia's economy. However, mining complexes without treatment facilities, such as those in Armenia, have adverse environmental impact. Moreover, soil contamination can pose a potential risk to human health, particularly, through the consumption of food crops. In this study, 12 soil and 32 vegetable composite samples were collected from the city of Kajaran where Armenia's biggest copper and molybdenum mine is located. The concentrations of Cu, Mo, Cd, Hg, As, and Pb were analyzed using atomic absorption spectrophotometer. Diet study was conducted using food frequency questionnaire. Non-carcinogenic and carcinogenic risks to human health through vegetable consumption were assessed. The results indicated that different vegetables have different trace element uptakes. Also, the transfer factors (TFs) for each vegetable varied across elements. TFs were less than 1 for the majority of trace elements. Nevertheless, in some samples of studied vegetables, the concentrations of Hg, Cd, and Pb exceeded the maximum allowable levels. THQ of Mo exceeded 1 for all the studied vegetables, while THQ of Cu exceeded 1 for potato and bean, indicating a potential health risk posed by chronic exposure. Exceedingly high levels of Mo exposure can be related to high incidence of anemia among Armenians, since Mo interacts with Cu and is a potential cause of copper deficiency-induced anemia. With regard to cancer risk, none of the carcinogenic risk values exceeded the threshold level.

Dietary Exposure Assessment of Potentially Toxic Trace Elements in Fruits and Vegetables Sold in Town of Kapan, Armenia.

Fruits and vegetables grown under the impact of Armenia's mining industry are widely sold in markets of adjacent towns. As the share of fruits and vegetables in Armenians' diet is significant, the present study aims to assess the dietary exposure of potentially toxic trace elements through the intake of fruits and vegetables sold in Kapan town, located in the biggest mining region of Armenia. The concentrations of Cu, Mo, Ni, Cr, Pb, Zn, Hg, As, and Cd in 15 types of fruits and vegetables were determined. Non-carcinogenic and carcinogenic risks were assessed. Although the estimated daily intakes of trace elements for each studied food item did not exceed health-based guidelines values, in case of the combined consumption of fruits and vegetables estimated cumulative daily intakes exceeded reference doses for Cu and Mo. Moreover, carcinogenic risk for the majority of fruits and vegetables exceeded the EPA recommended risk level of 10-6, indicating adverse health effect to local population. The outcomes of this study can serve as a basis for further research that will consider many other exposure pathways (i.e., inhalation or dermal pathways) in order to ensure the safety of the residents living under the impact of mining industry.