Biosafety measures for Alicyclobacillus spp. strains across various levels of biohazard.

Alicyclobacillus bacteria are important contaminants in the beverage industry because their spores remain in the product after usual pasteurization. At the same time, their impact on human health has yet to be characterized, as it is generally assumed to be low or non-existent. However, these bacteria are causing quality concerns mainly due to odor and taste changes of the product. Since potential health effects are not precisely known, an experimental assessment was performed, including a biosafety assessment of six viable and non-viable vegetative and spore forms of Alicyclobacillus spp. strains using cell cultures and rodent study. The monolayer of Caco-2 (Cancer coli-2) cells was investigated for its adsorption effect on the epithelium of the small intestine of mice. Lactate dehydrogenase leakage (LDH) and transepithelial electrical resistance (TEER) tests were used to ensure the integrity of the cell membrane and tight junctions. The methylthiazole tetrazolium bromide (MTT) assay examined in vitro cytotoxicity in Caco-2 and HepG2 cell lines. The hemolysis of erythrocytes was spectrophotometrically measured. The results showed negligible cytotoxicity or non-toxic response in mice. In conclusion, Alicyclobacillus spp. exhibited biocompatibility with negligible cytotoxicity and minimal safety concerns.

Designing a Multiepitope Vaccine against the Foodborne Pathogenic Bacteria Listeria monocytogenes Using Subtractive Immunoinformatics Approaches.

BACKGROUND: Listeria monocytogenes, a Gram-positive bacterium, is a prominent foodborne pathogen that causes listeriosis and poses substantial health hazards worldwide. The continuing risk of listeriosis outbreaks underlies the importance of designing an effective prevention strategy and developing a robust immune response by reverse vaccinology approaches. This study aimed to provide a critical approach for developing a potent multiepitope vaccine against this foodborne disease. METHODS: A chimeric peptide construct containing 5 B-cell epitopes, 16 major histocompatibility complex I (MHC-I) epitopes, and 18 MHC-II epitopes were used to create a subunit vaccination against L. monocytogenes. The vaccine safety was evaluated by several online methods, and molecular docking was performed using ClusPro to determine the binding affinity. Immune simulation was performed using the C-ImmSimm server to demonstrate the immune response. RESULTS: The results validated the antigenicity, non-allergenicity, and nontoxicity of the chimeric peptide construct, confirming its suitability as a subunit vaccine. Molecular docking showed a good score of 1276.5 and molecular dynamics simulations confirmed the construct's efficacy, demonstrating its promise as a good candidate for listeriosis prophylaxis. The population coverage was as high as 91.04% with a good immune response, indicating good antigen presentation with dendritic cells and production of memory cells. CONCLUSIONS: The findings of this study highlight the potential of the designed chimeric peptide construct as an effective subunit vaccine against Listeria, paving the way for future advances in preventive methods and vaccine design.

Monitoring and Health Risk Assessment of Lead and Cadmium in Date Palm Fruit Cultivars Growing in Morocco.

The present study aimed to investigate the levels of two toxic trace elements (lead (Pb) and cadmium (Cd)) in 84 date fruit samples of four varieties (Mejhoul, Bouittob, Aziza and Boufegouss) produced in four Moroccan areas, using graphite furnace atomic absorption spectrometry, and to assess their risk to the consumer health. Results showed that out of the total samples, Pb was detected in 39 date fruit samples (46.43%) in the range of 0.0001-0.118 mg/kg, with an average level of 0.016 ± 0.0005 mg/kg. For Cd levels, a total of 40 samples (47.62%) were found to be contaminated with Cd levels ranging from 0.0004 to 0.752 mg/kg, with a mean value of 0.079 ± 0.003 mg/kg. Cadmium levels exceeded the European maximum regulatory limit (MRL) of 0.020 mg/kg in 17 date samples, which represents 20.24%. However, only one sample (1.19%) was above the MRL for Pb (0.10 mg/kg) set by the Codex Alimentarius Commission (CAC). The hazard quotient (HQ) and hazard index (HI) values were found to be below 1. Furthermore, cancer risk (CR) levels exceeded the acceptable ranges. As a result, the non-carcinogenic risk assessments have concluded that consuming date fruits is safe and unlikely to harm harmful effects to the consumers. However, the date consumption may pose a significant lifetime carcinogenic risk to consumers from date-growing regions.

Whole Genome Analysis of Tibetan Kefir-Derived Lactiplantibacillus Plantarum 12-3 Elucidates Its Genomic Architecture, Antimicrobial and Drug Resistance, Potential Probiotic Functionality and Safety.

BACKGROUND: Lactiplantibacillus plantarum 12-3 holds great promise as a probiotic bacterial strain, yet its full potential remains untapped. This study aimed to better understand this potential therapeutic strain by exploring its genomic landscape, genetic diversity, CRISPR-Cas mechanism, genotype, and mechanistic perspectives for probiotic functionality and safety applications. METHODS: L. plantarum 12-3 was isolated from Tibetan kefir grains and, subsequently, Illumina and Single Molecule Real-Time (SMRT) technologies were used to extract and sequence genomic DNA from this organism. After performing pan-genomic and phylogenetic analysis, Average Nucleotide Identity (ANI) was used to confirm the taxonomic identity of the strain. Antibiotic resistance gene analysis was conducted using the Comprehensive Antibiotic Resistance Database (CARD). Antimicrobial susceptibility testing, and virulence gene identification were also included in our genomic analysis to evaluate food safety. Prophage, genomic islands, insertion sequences, and CRISPR-Cas sequence analyses were also carried out to gain insight into genetic components and defensive mechanisms within the bacterial genome. RESULTS: The 3.4 Mb genome of L. plantarum 12-3, was assembled with 99.1% completeness and low contamination. A total of 3234 genes with normal length and intergenic spacing were found using gene prediction tools. Pan-genomic studies demonstrated gene diversity and provided functional annotation, whereas phylogenetic analysis verified taxonomic identity. Our food safety study revealed a profile of antibiotic resistance that is favorable for use as a probiotic. Analysis of insertional sequences, genomic islands, and prophage within the genome provided information regarding genetic components and their possible effects on evolution. CONCLUSIONS: Pivotal genetic elements uncovered in this study play a crucial role in bacterial defense mechanisms and offer intriguing prospects for future genome engineering efforts. Moreover, our findings suggest further in vitro and in vivo studies are warranted to validate the functional attributes and probiotic potential of L. plantarum 12-3. Expanding the scope of the research to encompass a broader range of L. plantarum 12-3 strains and comparative analyses with other probiotic species would enhance our understanding of this organism's genetic diversity and functional properties.

AFM1 exposure in male balb/c mice and intervention strategies against its immuno-physiological toxicity using clay mineral and lactic acid bacteria alone or in combination.

CONTEXT: Aflatoxins are the most harmful mycotoxins that cause human and animal health concerns. Aflatoxin M1 (AFM1) is the primary hydroxylated metabolite of aflatoxin B1 and is linked to the development of hepatocellular carcinoma and immunotoxicity in humans and animals. Because of the important role of dairy products in human life, especially children, AFM1 is such a major concern to humans because of its frequent occurrence in dairy products at concentrations high enough to cause adverse effects to human and animal health. Reduced its bioavailability becomes a high priority in order to protect human and animal health. OBJECTIVES: This study aimed to investigate, in vivo, the ability of lactic acid bacteria (lactobacillus rhamnosus GAF01, LR) and clay mineral (bentonite, BT) mixture to mitigate/reduce AFM1-induced immunotoxicity, hepatotoxicity, nephrotoxicity and oxidative stress in exposed Balb/c mice. MATERIALS AND METHODS: The in vivo study was conducted using male Balb/c mice that treated, orally, by AFM1 alone or in combination with LR and/or BT, daily for 10 days as follows: group 1 control received 200 µl of PBS, group 2 treated with LR alone (2.108 CFU/mL), group 3 treated with BT alone (1 g/kg bw), group 4 treated with AFM1 alone (100 µg/kg), group 5 co-treated with LR + AFM1, group 6 co-treated with BT + AFM1, group 7 co-treated with BT + LR + AFM1. Forty-eight h after the end of the treatment, the mice were sacrificed and the blood, spleen, thymus, liver and kidney were collected. The blood was used for biochemical and immunological study. Spleen and thymus samples were used to thymocytes and splenocytes assessments. Liver and kidney samples were the target for evaluation of oxidative stress enzymes status and for histological assays. RESULTS: The results showed that AFM1 caused toxicities in male Blab/c mice at different levels. Treatment with AFM1 resulted in severe stress of liver and kidney organs indicated by a significant change in the biochemical and immunological parameters, histopathology as well as a disorder in the profile of oxidative stress enzymes levels. Also, it was demonstrated that AFM1 caused toxicities in thymus and spleen organs. The co-treatment with LR and/or BT significantly improved the hepatic and renal tissues, regulated antioxidant enzyme activities, spleen and thymus viability and biochemical and immunological parameters. LR and BT alone showed to be safe during the treatment. CONCLUSION: In summary, the LR and/or BT was able to reduce the biochemical, histopathological and immunological damages induced by AFM1 and indeed it could be exploited as one of the biological strategies for food and feedstuffs detoxification.

Biodiversity and biotechnological properties of lactic acid bacteria isolated from traditional Moroccan sourdoughs.

The present study aimed at characterizing lactic acid bacteria (LAB) strains isolated from traditional sourdoughs collected in different regions of Morocco. Isolated strains were firstly identified using Gram staining and catalase reaction test. Presumptive LAB strains were then checked for various phenotypical properties including growth at 45 °C, resistance to NaCl, enzyme production, acidification capacity, diacetyl and exopolysaccharide (EPS) production, and antifungal activity. Finally, selected LAB strains were identified using 16S rDNA sequencing. Results showed that 32.1% of the isolates were thermophilic (45 °C) and 83.9% were resistant to NaCl (6.5%). Moreover, 51.7 and 37.5% were able to produce diacetyl and EPS, respectively. Regarding enzyme production, 55.3 and 7.1% of the isolates showed lipolytic and proteolytic activities, respectively. Low pH values (3.37-3.76) were obtained after 24 h of incubation of LAB strains in de Man, Rogosa and Sharpe (MRS) broth. Antifungal activity test against Aspergillus flavus, Aspergillus niger and Penicillium spp. showed an inhibition rate up to 50%. Bacterial DNA sequencing showed that LAB isolates belong to seven species, chiefly Levilactobacillus brevis, Lentilactobacillus parabuchneri, Lactiplantibacillus plantarum, Pediococcus pentosaceus, Enterococcus hirae, Bifidobacterium pseudocatenulatum, and Companilactobacillus paralimentarius. These findings, for the first time in Moroccan sourdoughs, indicate that the isolated LAB strains have good multifunctional properties and could be suitable as good starters for sourdough bread production under controlled conditions.

Aflatoxins and Ochratoxin A in dried fruits from Morocco: Monitoring, regulatory aspects, and exposure assessment.

The present study aims to investigate the presence of Aflatoxins (AF) in 180 samples dried fruits and Ochratoxin A (OTA) in 210 samples dried fruits and grape juices collected in Morocco. Mycotoxins were analyzed by high performance liquid chromatography (HPLC) coupled to fluorescence detection and immunoaffinity columns (IAC) cleanup. Contamination levels were compared with the maximum regulatory limits (MRL) recently adopted in the country, and mycotoxin exposure of adult consumers was assessed. Results showed that 13.8% of samples were contaminated with AF, with incidences of 23.3, 23.3, 20, 13.8, and 3.3%, in raisins, figs, nuts, peanuts and pistachio, respectively. There were 12 samples (6.6%) that exceeded the MRL of 2-12 ng/g set for aflatoxin B1 (AFB1). While OTA was detected in 17.1% of samples, with incidences of 3.3, 3.3, 30, 30, and 53.3% in walnuts, pistachios, peanuts, raisins and figs, respectively, and a maximum value of 99.1 in dried raisins, that exceeded the MRL (10 ng/g) set for OTA. The co-occurrence of OTA and AF was observed in 4.7% of total samples. Dietary intake showed that the OTA exposure level was lower than safety guidelines set by The Joint FAO/WHO Expert Committee on Food Additives (JECFA) at 100 ng/kg b.w./week.

Comparative Study of Ochratoxin A Exposure through the Intake of Cereal Products in Two Climatic Moroccan Regions.

The present study aims to compare ochratoxin A (OTA) exposure through the intake of three cereal derivative products (bread, pasta and semolina) in two different Moroccan climatic regions (littoral and continental). OTA weekly intakes from cereal products were calculated using a deterministic approach for each region. Results showed a statistically significant difference (p < 0.05) of OTA exposure between the two regions. Indeed, the median OTA exposure was estimated at 48.97 ng/kg b.w./week in the littoral region, while it was estimated at 6.36 ng/kg b.w./week in the continental region. The probabilistic approach showed that, due to uncertainties, the 95th percentile of weekly OTA exposure associated with the three cereal products ranged from 66.18 to 137.79 (95% CI) with a median of 97.44 ng/kg body weight (b.w.)/week. Compared to the threshold of 100 ng/kg b.w./week, 95% of the cumulative distributions predicted an exceedance frequency between 0.42 and 17.30% (95% CI), with an exceedance frequency median of 4.43%. Results showed that cereal derivatives constitute an important vector of OTA exposure and cause a significant exceedance of toxicological reference value among large consumers in the littoral region, which suggests the urgency of reconsidering the maximum regulatory limit (MRL) set for OTA (3 µg/kg) in cereal derivatives by Moroccan authorities.

Prevention and Detoxification of Mycotoxins in Human Food and Animal Feed using Bio-resources from South Mediterranean Countries: a Critical Review.

Mycotoxins, which are natural toxic compounds produced by filamentous fungi, are considered major contaminants in the food and feed chain due to their stability during processing. Their impacts in food and feedstuff pollution were accentuated due the climate change in the region. They are characterized by their toxicological effects on human and animal health but also by their harmful economic impact. Mediterranean countries: Algeria, Egypt, Libya, Morocco and Tunisia are characterized by high temperatures and high relative humidity, particularly in littoral regions that provide favorable conditions for fungal growth and toxinogenesis. Many scientific papers have been published recently in these countries showing mycotoxin occurrence in different commodities and an attempt at bio-detoxification using many bio-products. In order to minimize the bioavailability and/or to detoxify mycotoxins into less toxic metabolites (bio-transforming agents), safe and biological methods have been developed including the use of lactic acid bacteria, yeasts, plant extracts and clays minerals from Mediterranean regions. The aim of this review is to present the pollution of mycotoxins in food and feedstuff of humans and animals and to discuss the development of effective biological control for mycotoxin removal/detoxification and prevention using bio-products. This review will also elucidate the new used natural products to be considered as a new candidates for mycotoxins detoxification/prevention on animal feedstuffs.

Occurrence of pre- and postharvest multi-mycotoxins in durum wheat grains collected in 2020 and 2021 in two climatic regions of Tunisia.

Twenty two mycotoxins in 136 durum wheat collected from Tunisia in 2020 and 2021 were investigated. Mycotoxins were analyzed by UHPLCMS/MS. In 2020, 60.9% of the samples were contaminated with Aflatoxin B1 (AFB1) and/or enniatin. Whereas, in 2021, 34.4% were contaminated by enniatins. AFB1 was detected only in 2020, in the continental region (6/46) and all samples exceeded limits. AFB1 was detected in stored wheat (24-37.8 µg/kg) but also in pre-stored wheat (17-28.4 µg/kg) and in one sample collected in the field (21 µg/kg). Enniatin A1, enniatin B and enniatin B1 were detected in wheat collected in the field (30-7684 µg/kg), pre-storage (42-1266 µg/kg) and storage (65.8-498.2 µg/kg) from the continental region also, in sample collected in pre-storage (31.3-1410 µg/kg) and at harvest (48- 1060 µg/kg). Samples had a water activity less than 0.7 and moisture content ranged between 09-14%. AFB1 level represent a health risk to the Tunisian consumers.

Trace elements in Foodstuffs from the Mediterranean Basin-Occurrence, Risk Assessment, Regulations, and Prevention strategies: A review.

Trace elements (TEs) are chemical compounds that naturally occur in the earth's crust and in living organisms at low concentrations. Anthropogenic activities can significantly increase the level of TEs in the environment and finally enter the food chain. Toxic TEs like cadmium, lead, arsenic, and mercury have no positive role in a biological system and can cause harmful effects on human health. Ingestion of contaminated food is a typical route of TEs intake by humans. Recent data about the occurrence of TEs in food available in the Mediterranean countries are considered in this review. Analytical methods are also discussed. Furthermore, a discussion of existing international agency regulations will be given. The risk associated with the dietary intake of TEs was estimated by considering consumer exposure and threshold values such as Benchmark dose lower confidence limit and provisional tolerable weekly intake established by the European Food Safety Authority and the Joint FAO/WHO Expert Committee on Food Additives, respectively. Finally, several remediation approaches to minimize TE contamination in foodstuffs were discussed including chemical, biological, biotechnological, and nanotechnological methods. The results of this study proved the occurrence of TEs contamination at high levels in vegetables and fish from some Mediterranean countries. Lead and cadmium are more abundant in foodstuffs than other toxic trace elements. Geographical variations in TE contamination of food crops clearly appear, with a greater risk in developing countries. There is still a need for the regular monitoring of these toxic element levels in food items to ensure consumer protection.

Ochratoxin a in breast milk in Morocco: the affecting dietary habits of the lactating mothers and the degree of exposure of newborns "CONTAMILK study".

The aim of the present study is to evaluate the level of contamination of breast milk (BM) by ochratoxin A, among Moroccan lactating mothers in the city of Rabat, and to identify the associated factors of exposure, also to estimate the degree of exposure of the breastfeed infant. The analysis of ochratoxin A (OTA) was accomplished by ELISA method on 82 colostrum samples. OTA was detectable (>0.08 ng/mL) in 55% of samples with a maximum concentration of 10.04 ng/mL, and the levels exceeded 0.5 ng /mL in 50 % of the samples. In addition, several factors and dietary habits affect significantly the level of OTA in the analyzed samples of breast milk including, the consumption of industrial dairy products, the frequency of consumption of canned foods, dried fruits and legumes, also the period of breast milk collection. Besides, OTA was higher than the tolerable daily intake for 49% newborns. However, these results need to be confirmed by multicenter studies to more broadly estimate the levels of exposition of Moroccan population to OTA. Furthermore, awareness campaigns are recommended to inform the public, especially pregnant women and lactating women about appropriate preventive measures to limit exposure to this mycotoxin.

Aflatoxin M1 in Africa: Exposure Assessment, Regulations, and Prevention Strategies - A Review.

Aflatoxins are the most harmful mycotoxins causing health problems to human and animal. Many acute aflatoxin outbreaks have been reported in Africa, especially in Kenya and Tanzania. When ingested, aflatoxin B1 is converted by hydroxylation in the liver into aflatoxin M1, which is excreted in milk of dairy females and in urine of exposed populations. This review aims to highlight the AFM1 studies carried out in African regions (North Africa, East Africa, West Africa, Central Africa, and Southern Africa), particularly AFM1 occurrence in milk and dairy products, and in human biological fluids (breast milk, serum, and urine) of the populations exposed. Strategies for AFM1 detoxification will be considered, as well as AFM1 regulations as compared to the legislation adopted worldwide and the assessment of AFM1 exposure of some African populations. Egypt, Kenya, and Nigeria have the highest number of investigations on AFM1 in the continent. Indeed, some reports showed that 100% of the samples analyzed exceeded the EU regulations (50 ng/kg), especially in Zimbabwe, Nigeria, Sudan, and Egypt. Furthermore, AFM1 levels up to 8,000, 6,999, 6,900, and 2040 ng/kg have been reported in milk from Egypt, Kenya, Sudan, and Nigeria, respectively. Data on AFM1 occurrence in human biological fluids have also shown that exposure of African populations is mainly due to milk intake and breastfeeding, with 85-100% of children being exposed to high levels. Food fermentation in Africa has been tried for AFM1 detoxification strategies. Few African countries have set regulations for AFM1 in milk and derivatives, generally similar to those of the Codex alimentarius, the US or the EU standards.

Multi-mycotoxin contamination of green tea infusion and dietary exposure assessment in Moroccan population.

Green tea infusion is one of the most widely drunk beverages worldwide due to its health benefits associated with microelements, essential oils, and polyphenols, etc. Several studies have reported that green tea is subjected to contamination by various toxigenic fungi. Thus, this work aims to investigate the co-occurrence of 15 mycotoxins [four aflatoxins (AFB1, AFB2, AFG1, AFG2), ochratoxin A (OTA), beauvericin (BEA), four enniatins (ENA, ENA1, ENB, ENB1), zearalenone (ZEN), alternariol (AOH), tentoxin (TENT), T-2 and HT-2 toxins] in green tea samples available in Morocco by liquid chromatography tandem mass spectrometry method. Analytical and consumption data were then used to assess the dietary exposure for the population. Out of 111 total green tea samples, 62 (56%) were contaminated by at least one mycotoxin. The most found mycotoxins in samples were AOH (40%), ZEN (35%), AFG1 (2%), AFB2 (2%), ENB (2%) and TENT (1%). The highest level was found for ZEN with 45.8 ng/g. There is no sample that exceeded the recommended levels set by European Pharmacopoeia for certain mycotoxins in plant material. Although multi-mycotoxin co-occurred in samples (33%), the probable estimated daily intake values show that the intake of mycotoxins through the consumption of green tea does not represent a risk for the population.

Occurrence of Free and Conjugated Mycotoxins in Aromatic and Medicinal Plants and Dietary Exposure Assessment in the Moroccan Population.

Aromatic and medicinal plants (AMPs), as herbal material, are subjected to contamination by various mycotoxin-producing fungi, either free and conjugated. Such a problem is associated with poor storage practices, and lack of adopting good agricultural practices and good harvesting practices. Nevertheless, AMPs are poorly investigated. The purpose of this study was to investigate the co-occurrence of 15 mycotoxins (four aflatoxins (AFB1, AFB2, AFG1, and AFG2), ochratoxin A (OTA), beauvericin (BEA), four enniatins (ENA, ENA1, ENB, and ENB1), zearalenone (ZEN), alternariol (AOH), tentoxin (TENT), T-2, and HT-2 toxins) in 40 samples of AMPs frequently consumed in Morocco by using liquid chromatography tandem mass spectrometry. Evaluation of conjugated mycotoxins and their identification using liquid chromatography coupled to time-of-flight mass spectrometry with ion mass exact was also carried out. Results showed that 90% of the analyzed samples presented at least one mycotoxin, and 52% presented co-occurrence of them. Mycotoxins detected were: AOH (85%), ZEN (27.5%), ß-ZEL (22%), AFG1 (17.5%), TENT (17.5%), ENB (10%), AFG2 (7.5%), α-ZEL (5%), ENA1 (2.5%), and HT-2 (2.5%), while the conjugated mycotoxins were ZEN-14-Glc (11%) and ZEN-14-Sulf (9%). The highest observed level was for AOH, with 309 ng/g. Ten samples exceeded the recommended levels set by the European Pharmacopoeia for AF mycotoxins in plant material (4 ng/g), and three samples exceeded the maximum limits for AFs (10 ng/g) in species established by the European Commission. Although the co-occurrence of several mycotoxins in AMP samples was observed, the dietary exposure assessment showed that the intake of mycotoxins through the consumption of AMP beverages does not represent a risk for the population.

Trace Elements in Breakfast Cereals and Exposure Assessment in Moroccan Population: Case of Lead and Cadmium.

The role of breakfast cereals in a balanced diet has been recognized for many years. Such foods should be safe and not contain toxic substances, especially trace elements. Among these elements, lead (Pb) and cadmium (Cd) are two important inorganic food contaminants. In this study, we assessed the contamination levels of breakfast cereal samples available in Morocco with Pb and Cd. For this, a total of sixty-two (n = 62) samples of breakfast cereals purchased in different markets in the country were surveyed for their Pb and Cd contents by using atomic absorption spectrophotometer (GF-AAS) after total mineralization of samples. Results showed that out of 62 total samples, 47 samples (75.8%) were contaminated with Pb concentrations in the range of 0.016-1.057 µg/g. The remaining samples (24.2%) were under the detection limit (LOD) of Pb. In the case of Cd, 41 samples (66.1%) were contaminated with Cd levels that ranged between 0.011 and 0.123 µg/g. In the present study, four samples (6.45%) of breakfast cereals are above the maximum limit (0.2 µg/g) set by the European Commission Regulation No 1881/2006 for Pb in cereals. However, for the Cd, only one sample exceeded the maximum limit set for this element (0.1 µg/g). The levels compare well with those reported worldwide for similar foodstuffs. The estimation of the provisional weekly intakes of the two elements (Pb and Cd) showed that the risks of development of toxicological effects through breakfast cereals are very low. However, it is important that the long-term exposure to these elements be kept to minimum. This is the first study on the co-occurrence of the two trace elements (Pb and Cd) in breakfast cereal samples commercialized in Morocco.

Exposure assessment to ochratoxin A through the intake of three cereal derivatives from the Moroccan market.

Previous studies reported the contamination of cereals products with OTA in Morocco. Given bread, pasta and semolina are staple consumed food in the country, this study aims to assess the OTA exposure levels for the adult consumers in the country through cereal derivatives intake, by using a deterministic model based on the crossover of consumption and contamination data of 457 cereals products samples. The study also set out to determine under what conditions the OTA contributions from these 3 cereal derivatives alone were enough to cause a real public health problem in the country. The Estimated Weekly Intake EWI (ng/kg b.w./week) was used as an indicator of exposure for each cereal derivative. The global exposure (Total EWI) was determined by the summation of the EWI obtained for each cereal derivative for different consumers groups and at different OTA levels and nine scenarios were elaborated. Results showed that the total average OTA intake from the three cereal derivatives was estimated at 13.5 ng/kg b.w./week. Despite its low contamination levels, bread has been shown to be the main vector of exposure to OTA because of the high consumption levels by Moroccan adults. Its contribution to the total OTA intake was between 81 and 84% depending on the scenario. The contribution of the other investigated cereal products to total OTA intake seems to be modest and does not exceed 10% and 7% for semolina and pasta, respectively. The predominance of cereal derivatives in Moroccan food thus constitutes a potential risk factor for OTA exposure. Indeed, the risk of occurrence of potential adverse effects of OTA is real in three scenarios discussed in the study since the PTWI established by JECFA (100 ng/kg b.w./week) was exceeded. Moreover, the Maximum limit (3 ng/g) set by Moroccan regulations in 2016, seems to be less protective for the Moroccan consumers compared to other countries due to the high consumption level of cereal products. This situation is likely to cause a public health problem by the occurrence of specific pathologies at highly OTA exposed adults, especially high bread consumers.

Aflatoxin M1 prevalence in breast milk in Morocco: Associated factors and health risk assessment of newborns "CONTAMILK study".

The aim of this study was to assess the contamination of breast milk by aflatoxin M1 among nursing mothers from Rabat, Morocco, and to explore its association with several maternal parameters and dietary habits. In addition, the health risk assessment of the newborns by the estimation of the daily intake. A competitive Enzyme Linked Immunosorbent Assay method was used for the analysis of aflatoxin M1 in breast milk samples. Analytical results indicate that out of 82 total samples, 43 samples (52.4%) of milk were positive. Aflatoxin M1 levels ranged from undetectable to 13.33 ng/L, while the mean level was 5.75 ± 3.44 ng/L. Besides, several factors and foodstuffs seem to increase the level of AFM1 in breast milk. As regards the estimated daily intake of aflatoxin M1, it varies between immeasurable and a maximum of 1.16 ng/kg.bw. The degree of exposure to AFB1 and the levels of its metabolite AFM1 in breast milk were low, compared to some studies from other countries. Further investigations and periodic monitoring programs are recommended in large samples and in many cities of morocco to assess the level of exposure of the Moroccan population.

Lactobacillus plantarum MON03 counteracts zearalenone génotoxicty in mice: Chromosome aberrations, micronuclei, DNA fragmentation and apoptotique gene expression.

Zearalenone (ZEN) is a potent estrogenic metabolite produced by some Fusarium species. No treatment has been successfully employed to get rid against ZEN contained in foods and/or mitigates its genotoxicity. This study was conducted to evaluate the ability of lactic acid bacteria, isolated from Tunisia traditional butter, Lactobacillus plantarum MON03 (LP) to protect mice against cytotoxicity and genotoxicity induced by ZEN. Two doses of LP (2 × 109 CFU/L, ∼2 mg/kg and 4 × 109 CFU/L, ∼4 mg/kg) was added alone or in combination with a toxic intragastric ZEN (40 mg/kg representing 8% of LD50) dose daily for 2 wk by oral gavage. The control group received distilled water. The positive control groups received Colchicin (4 mg/kg bw) for the micronucleus assay and mitomycin C (1 mg/kg bw) for the chromosome aberrations assay. 48 h after treatment, the small intestines, femur and tibia are dissected out. Small intestines were collected for the determination of DNA fragmentation, genes expression and target proteins content. The results show that ZEN was cytotoxic and genotoxic to mice as indicated by the increase in frequencies of polychromatic erythrocytes micronucleated (PCEMN) and chromosomal aberrations in bone marrow cells. In the small intestine ZEN was increased DNA fragmentation, down regulated the expressions of caspase-3, caspase-9, and Bax as well as up-regulated the expression of Bcl-2 and their target proteins. The simultaneous intragastric administration of LP with ZEN resulted in a decrease of PCEMN number and chromosomal aberrations frequency and in an increase of polychromatic erythrocytes (PCE) in bone marrow cells compared with the group treated with ZEN alone. In addition, LP succeeded to alleviate the disturbances in DNA fragmentation and the expression of these genes and their target proteins. It could be concluded that the use of LP induced protective effects against genotoxicity of ZEN in part through adhesion and so likely diminished its bio-availability in gastro-intestinal tract.

Multi-Occurrence of Twenty Mycotoxinsin Pasta and a Risk Assessment in the Moroccan Population.

In the present study, the multi-occurrence of twenty (20) mycotoxins in pasta samples consumed in Morocco was assessed. For this, a modified Quick, Easy, Cheap Effective, Rugged, and Safe method was validated. The mycotoxins studied were identified and quantified by liquid chromatography⁻tandem mass spectrometry (LC⁻MS/MS) and gas chromatography⁻tandem mass spectrometry (GC-MS/MS). The validated method was applied to one hundred and six (n = 106) pasta samples purchased from several areas in the country. The analytical results showed that 99 out of 106 total samples (93.4%) were contaminated with at least one mycotoxin. Nine mycotoxins (Aflatoxin B1, Enniatin B, Enniatin B1, Enniatin A1, Zearalenone, Deoxynivalenol, 3-Acetyl-Deoxynivalenol, T-2, and HT-2 toxins) were present in the pasta samples. Enniatin B and Enniatin B1 were the predominant mycotoxins. The Zearalenone, Deoxynivalenol, HT-2, and T-2 toxins were present in 51.8%, 43.5%, 34.9%, and 16% of samples, respectively. Aflatoxin B1 was detected in only 2 samples. Risk exposure assessment concluded that mycotoxin levels found in pasta do not pose a significant human health risk for the Moroccan population. This is the first paper drafted on the multi-occurrence of mycotoxins in pasta from this country.