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.

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.

Aspergillus section Flavi and aflatoxins in dried figs and nuts in Algeria.

The presence of Aspergillus section Flavi and aflatoxin (AF) contamination was investigated in 112 samples of peanuts, almonds and dried figs collected in Algeria. The occurrence of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) in different commodities has been determined with a sensitive method based on high performance liquid chromatography (HPLC) coupled with fluorescence detection with post-column photochemical derivatisation. Analytical results indicated that 28 samples of peanuts, 16 samples of almonds and 26 samples of dried figs contained detectable levels of AFs. A total of 69 samples (61.6%) were contaminated with AFB1 ranging from the limit of quantification to 174 µg kg-1. AFB2 was found in 12 samples (10.7%) and varied from 0.18 to 193 µg kg-1. Seven samples revealed AF concentrations lower than the limit of quantification. Eleven peanut and fourteen dried fig samples exceeded the European maximum limits for AFB1.

Antifungal properties of an actinomycin D-producing strain, Streptomyces sp. IA1, isolated from a Saharan soil.

An actinomycete strain named IA1, which produced an antimicrobial compound, was isolated from a Saharan soil in In Amenas, Algeria. The study of the 16S rDNA sequence of this strain permitted to relate it to Streptomyces mutabilis NBRC 12800(T) (99.93% of similarity). Strain IA1 exhibited strong activity against a wide range of plant pathogenic fungi. One bioactive compound produced in large amounts (46.7 mg L(-1) day(-1) ), named YA, was isolated and purified by TLC and reverse phase HPLC. The structure elucidation of the pure substance, using combined data from UV visible, NMR spectra, and mass spectrometry, permitted to identify it as actinomycin D, and was thus found for the first time in S. mutabilis related species. The biocontrol abilities of the strain IA1 and compound YA were evaluated through two diseases, i.e., chocolate spot of field bean and Fusarium wilt of flax. The occurrence of the two fungal diseases was effectively reduced. The reduction of chocolate spot disease symptoms reached 80 and 91.7% with IA1 and YA seedlings pretreatments, respectively. Soil pretreatment with IA1 or YA also allowed to reduce Fusarium wilt disease impact by almost 60%.

Streptomonospora algeriensis sp. nov., a halophilic actinomycete isolated from soil in Algeria.

A halophilic actinomycete strain, designated H27(T), was isolated from a soil sample collected from a hypersaline habitat in Djelfa Province (North-Central Algeria), and then investigated using a polyphasic taxonomic approach. The strain was observed to produce poor aerial mycelium, which formed short chains of oval to cylindrical-shaped spores at maturity, and non fragmented substrate mycelium. The optimum NaCl concentration for growth was found to be 10-15 % (w/v) and the optimum growth temperature and pH were found to be 28-37 °C and 6-7, respectively. The diagnostic diamino acid in the cell-wall peptidoglycan was identified as meso-diaminopimelic acid. The predominant menaquinones of strain H27(T) were identified as MK-11 (H4) and MK-10 (H6). The major fatty acids were found to be iso-C16:0, anteiso-C17:0, 10 methyl C17:0 and 10 methyl C16:0. The diagnostic phospholipids detected were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and phosphatidylinositol. The chemotaxonomic properties of strain H27(T) are consistent with those shared by members of the genus Streptomonospora. 16S rRNA gene sequence analysis indicated that strain H27(T) is most closely related to Streptomonospora alba DSM 44588(T) (98.8 %) and Streptomonospora flavalba DSM 45155(T) (98.7 %) whereas the DNA-DNA relatedness values between strain H27(T) and the two type strains were 17.1 and 57.9 %, respectively. Based on the combined genotypic and phenotypic evidence, it is proposed that strain H27(T) should be classified as representative of a novel species, for which the name Streptomonospora algeriensis sp. nov. is proposed. The type strain is H27(T) (=DSM 45604(T) =CCUG 63369(T) =MTCC 11563(T)).

Aspergillus section Flavi and aflatoxins in Algerian wheat and derived products.

Wheat and its derivatives are a very important staple food for North African populations. The aim of this study was to analyze populations of Aspergillus section Flavi from local wheat based on aflatoxins (AFs), cyclopiazonic acid (CPA) and sclerotia production, and also to evaluate AFs-contaminated wheat collected from two different climatic regions in Algeria. A total of 108 samples of wheat were collected during the following phases: pre-harvest, storage in silos and after processing. The results revealed that among the Aspergillus species isolated, those belonging to section Flavi were predominant. Of the 150 strains of Aspergillus section Flavi isolated, 144 were identified as Aspergillus flavus and 6 as Aspergillus tamarii. We showed that 72% and 10% of the A. flavus strains produced AFs and CPA, respectively. Among the 150 strains tested, 60 produced amounts of AFB1 ranging from 12.1 to 234.6 microg/g of CYA medium. Also, we showed that most strains produced large sclerotia. AFB1was detected by HPLC in 56.6% of the wheat samples and derived products (flour, semolina and bran) with contamination levels ranging from 0.13 to 37.42 microg/kg.