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.

The ameliorative effect of Lactobacillus paracasei BEJ01 against FB1 induced spermatogenesis disturbance, testicular oxidative stress and histopathological damage.

Fumonisin B1 (FB1) is a possible carcinogenic molecule for humans as classified by the International Agency for Research on Cancer (IARC) in 2B group. In livestock, it is responsible for several mycotoxicoses and economic losses. Lactobacillus strains, inhabitants of a wide range of foodstuffs and the gastrointestinal tract, are generally recognized as safe (GRAS). Thus, the aim of this work was to evaluate the protective effect of Lactobacillus paracasei (LP) against FB1-induced reprotoxicities including testicular histopathology, sperm quality disturbance, and testosterone level reduction.Pubescent mice were divided randomly into four groups and treated for 10 days. Group 1: Control; Group 2: FB1 (100 µg/kg b.w); Group 3: LP (2 × 109 CFU/kg b.w); Group 4: LP (2 × 109 CFU/kg b.w) and FB1 (100 µg/kg b.w). After the end of the treatment, animals were sacrificed. Plasma, epididymis, and testis were collected for reproductive system studies.Our results showed that FB1 altered epididymal sperm quality, generated oxidative stress, and induced histological alterations. Interestingly, these deleterious effects have been counteracted by the LP administration in mice.In conclusion, LP was able to prevent FB1-reproductive system damage in BALB/c mice and could be validated as an anti-caking agent in an animal FB1-contaminated diet.

Potential protective effect of lactic acid bacteria against zearalenone causing reprotoxicity in male mice.

Zearalenone (ZEN) is a worldwide fusarotoxin that poses a threat to the consumer due to its chronic toxicity. Herein we examined the effects of ZEN on adult mouse testis, focusing on oxidative stress, biochemical and morphological parameters. In addition, since cytoskeletal remodeling is a key event for the production of good quality gametes, the expression and localization of two proteins, Dishevelled-associated activator of morphogenesis 1 (DAAM1) and Prolyl endopeptidase (PREP), involved in cytoskeletal dynamics during spermatogenesis were evaluated. To ameliorate the testicular dysfunction induced by ZEN we tested the eventual protective effects of lactic bacteria Lactobacillus plantarum MON03 (LP) on its reprotoxicity. Adult male mice were then treated daily for 2 wks by oral gavage with ZEN and/or LP. The results confirmed that ZEN altered sperm parameters, generated oxidative stress and provoked structural alteration, evidenced by the increased number of abnormal seminiferous tubules and of apoptotic cells, particularly Leydig cells. Interestingly, at molecular level we evaluated, for the first time, the ability of ZEN to alter DAAM1 and PREP protein level and localization. Moreover, the co-treatment with LP, thanks to its capacity to reduce ZEN bioavailability in the gastrointestinal tract, ameliorated all the considered parameters. These results suggest the use of this probiotic as food supplement to prevent/counteract ZEN-induced reprotoxicity.

Mycotoxins occurrence in milk and cereals in North African countries - a review.

North African countries; Algeria, Egypt, Libya, Morocco and Tunisia suffer from mycotoxin contamination. Various studies have indicated the presence of mycotoxins in raw milk and cereals (i.e. wheat, barley, maize and cereal-based products). Aflatoxins (AFs), Aflatoxin M1 (AFM1), Ochratoxin A (OTA), Fumonisin (FB1) and Zearalenone (ZEN)-mycotoxin are the most detected due to climatic change in the region. In this review, we will present the kind of foods and feeds cereals and milk based products contaminated and the level of their contaminated mycotoxin. On the other hand, researchers try to find biologic methods to remove/mitigate mycotoxins in food and feed using bio-products. But the research works concerning legislations and mycotoxin risk assessment still rare. Therefore, it appears necessary to make review on the current status of mycotoxins in North African countries in order to explore data related to contamination of basic food in this region and to highlight the problem to the policy-makers to establish a serious legislation on this matter. On the other hand, to give more information to the worldwide readers about the impact of climate change on the food and feed pollution on mycotoxins in the Mediterranean Sea region.

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.

Efficacy of lactic acid bacteria supplementation against Fusarium graminearum growth in vitro and inhibition of Zearalenone causing inflammation and oxidative stress in vivo.

Fusarium graminearum invasion and Zearalenone (ZEN)-mycotoxin contamination are considered the most global threat to food and feed. This study investigates the effect Lactobacillus plantarum MON03 viable cells (LPVC) and LP free cells supernatant (LPFCS) against Fusarium graminearum growth and ZEN production in vitro and evaluates if treatment with LP viable cells can counteract the negative effect of ZEN on inflammation and oxidative stress in mesenteric lymph nodes and serum biochemical parameters in mice. For the in vitro study, 7 days of LPVC, LPFCS and F. graminearum co-incubation at different concentrations was done in order to determine the antifungal activity and ZEN- production inhibition. Regarding the in vivo study, Balb/c mice were treated as following: Control, ZEN group, LP group and ZEN + LP group for 30 days. In vitro, LPVC showed an excellent antifungal activity after 7 days of co-incubation (103 CFU/ml). LPVC was succeeded also to inhibit ZEN production by the fungi. In vivo, ZEN has shown an important oxidative damage. As a result of the exposure to ZEN, an increase cytokines, as effectors of an inflammatory response, were observed in the mesenteric lymph nodes (MLN) of intoxicated mice. In parallel, a serum biochemical change was also observed. LPVC induced a reduction of ZEN-induced oxidative stress and counteracts also the biochemical parameters damage and the inflammatory markers increased by ZEN. LPVC can be valorized as an anti-cating agent in the vitro and in the gastro-intestinal tract to decrease ZEN-toxic effects.

Immunological effects of AFM1 in experimental subchronic dosing in mice prevented by lactic acid bacteria.

AIM: Recently, higher contamination by aflatoxin M1 (AFM1) has been detected in many countries. Unfortunately, many tons of contaminated milk and milk byproducts are removed from the food chain to avoid human contamination; as a consequence of higher economic losses. Fewest number of studies are interested to AFM1 detoxification using lactic acid bacteria. MATERIALS AND METHODS: In this study, AFM1-degradation using Lactobacillus paracasei BEJ01 (LPBEJ01) was tested in vitro. The preventive effect of LPBEJ01 against AFM1 immunobiological effects in mice are treated orally during 3 weeks with 100 µg AFM1, LPBEJ01 (2 × 109 CFU/ml∼2 mg/kg p.c.) and a mixture of AFM1 and LPBEJ01. RESULTS: In vitro LPBEJ01 was found able to absorb 98% of AFM1 (100 µg/ml) in liquid medium after 24 h and more than 95% of AFM1 could be eliminated after 24 h in a solid-state fermentation. Animals treated with AFM1 obtained lower body weight than the control ones. The mitogenic response of spleen mononuclear cells (SMCs) in vivo was higher in mice treated with AFM1. The SMC of mice treated with AFM1 produced lower levels of IL-2, higher levels IL-4 and no effect on IL-10 production. The peritoneal macrophages of mice that treated with AFM1 released less H2O2, while mice exposed orally with the mixture of AFM1 and LPBEJ01 produced higher levels. CONCLUSION: LPBEJ01 was safe and it did not have any sign of toxicity. It can be used as an additive for AFM1-detoxification contamination in the food chain in countries suffering from this problem.

Lactobacillus paracasei alleviates genotoxicity, oxidative stress status and histopathological damage induced by Fumonisin B1 in BALB/c mice.

Fumonisin B1 (FB1) is a prevelant mycotoxin in our alimentary chain. It was produced by the fungi of the genus Fusarium verticillioides and Fusarium proliferatum. FB1 was playing as a competitive inhibitor of ceramide synthase; a key enzyme in the biosynthesis of sphingolipids. Indeed, it was associated with several affects in humans and livestock animals. The aim of our report was elucidated to evaluate the protective effects of Lactobacillus paracasei BEJ01 (LP) isolated from the traditional butter of Tunisia against the FB1 genotoxicity, hematoxicity, oxidative stress and histological damage in liver and kidney of BALB/c mice. Forty old week mice were randomly divided into four treatment groups (10 mice/group): Group 1: control; Group 2: LP (2 × 109 CFU/ml ~ 2 mg/kg p.c); Group 3: FB1 (100 µg/kg p.c.); Group 4: LP (2 × 109 CFU/ml ~ 2 mg/kg p.c) + FB1 (100 µg/kg p.c.). 48 h after the end of the treatment (10 days), the mice were sacrificed and the blood, liver and kidney were collected. The blood was used for hematological and biochemical studies. Liver and kidney samples were the target for evaluation of oxidative stress enzymes status and histopathological damage. The results indicated that FB1 was induced toxicities in the blood parameters and in liver and kidney tissues as well as in the profile of oxidative stress enzymes levels. The co-treatment with LP was found able to mitigate the FB1 toxicities by reduction of its bioavailability in the gastro intestinal tract. However, treatment with LP alone was safe and no sign of toxicity was showed. In Summary, the LP strain was able to prevent FB1 toxicities and indeed it could be exploited as one of the biological strategies for foodstuffs decontamination.

Zearalenone nephrotoxicity: DNA fragmentation, apoptotic gene expression and oxidative stress protected by Lactobacillus plantarum MON03.

The present study was conducted to determine the abilities of the living Lactobacillus plantarum MON03 cells to degrade Zearalenone (ZEN) in liquid medium, and to elucidate the preventive effect in ZEN-contaminated balb/c mice showing kidney damage. The DNA fragmentation, Bcl-2 and Bax gene expression, caspase-3 activity, mRNA level of inflammation-regulating cytokines and histology of kidney tissues were examined. Female Balb/c mice were divided into four groups (10/group) and treated daily for 2 wk by oral gavage with lactic acid bacteria (L. plantarum MON03) 2 × 109 CFU/L, ~2 mg/kg only, ZEN (40 mg/kg BW) only, ZEN (40 mg/kg BW) + lactic acid bacteria (L. plantarum MON03, 2 × 109 CFU/L, ~2 mg/kg). Control group received vehicle. At the end of experiment, the kidney was collected for the determination of DNA fragmentation, Bcl-2 and Bax gene expression,caspase-3 activity, Malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) content, as well as for any alterations in expression of total antioxidant activity (TAC) and mRNA levels of inflammation-regulating cytokines (e.g., IL-10, IL-6, TNF-alpha). The results indicated that, kidney cells exposure to ZEN led to increased caspase-3 activity, MDA, and IL-10, IL-6, TNF-alpha and Bax mRNA levels, but decreased TAC content and down-regulated expression of GSH-Px and CAT and Bcl-2 mRNA. Co-treatment with ZEN plus LP suppressed the levels of DNA fragmentation; normalized kidney MDA and increased CAT levels, up-regulated expression of GSH-Px and CAT, and normalized mRNA levels of the analyzed cytokines. It's concluded that ZEN might have toxic effects in kidney. Further, it can be seen that use of LP induced protective effects against the oxidative stress and kidney toxicity of ZEN in part through adhesion (and so likely diminished bioavailability).

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.

Review: Biotechnology of mycotoxins detoxification using microorganisms and enzymes.

Mycotoxins are secondary metabolites produced by various toxigenic fungi belonging to the genera Aspergillus, Fusarium and Penicillium. They have mutagenic, teratogenic, carcinogenic, immunosuppressive and endocrine disrupting effects on both humans and animals. The ubiquitous occurrence and heat stability of these mycotoxins during food and feed processing pose serious health problems in human and animals. Therefore successful biotechnological detoxification tools are in great demand. Recently, the use of bacteria, yeast, moulds and enzymes to mitigate/bind or biodegrade various mycotoxins is a novel strategy used for mycotoxins removal. In this review, recent development in biological control of mycotoxins removal and detoxification will be discussed.

Lactobacillus plantarum alleviate aflatoxins (B1 and M1) induced disturbances in the intestinal genes expression and DNA fragmentation in mice.

This study aimed to assess the disturbances in intestinal genes expression and DNA fragmentation in mice treated orally with aflatoxin B1 (AFB1) or aflatoxin M1 (AFM1) and the protective activity of Lactobacillus plantarum (LP). Male Balb/c mice were divided into 6 groups including the control group, the group treated with 2 mg/kg b.w of LP (2 × 109 cfu/mL), the groups treated with AFB1 or AFM1 (100 µg/kg b.w), and the groups treated with AFB1 or AFM1 during, after or before LP. Small intestines were collected for the determination of DNA fragmentation, gene expression and target protein content. The results showed that AFB1 or AFM1 increased DNA fragmentation, down regulated the expressions of caspase-3, caspase-9, CYP3A13, Bax and p53 as well as up-regulated the expression of TNF-α and Bcl-2 and their target proteins. LP succeeded to alleviate the disturbances in DNA fragmentation and the expression of these genes. The improvement was more pronounced in the group co-administered with the toxins plus LP. It could be concluded that AFB1 and AFM1 induced disturbances in intestinal function via the disturbances in DNA fragmentation and genes expression. LP induced a potential protective effect and is considered a promising agent against the genotoxicity induced by these mycotoxins.

Immuno-physiological alterations from AFB1 in rats counteracted by treatments with Lactobacillus paracasei BEJ01 and montmorillonite clay mixture.

High contamination by aflatoxin B1 (AFB1) has been detected in Beja province (Tunisia) in many dairy products and animal feed, which has resulted in many tons of cereals and cereals being removed from the market, causing economic loss. While removal represents a means of reducing risk, exposures still occur. Studies have increasingly focused on means of AFB1 biodegradation/elimination using lactic acid bacteria and clay mineral. In the study here, Lactobacillus paracasei BEJ01 (LP) and montmorilonite clay (MT) were used to reduce the physio-/immunotoxicologic disorders that could develop in rats that underwent AFB1 exposures for a total of 7 consecutive days. The results indicated that rats treated with AFB1 (80 µg/kg BW) alone had significant decreases in lymphocytes in their blood (including B-lymphocytes, CD3(+), CD4(+), and CD8(+) T-lymphocyte subtypes, and NK cells), immunoglobulins (IgA and IgG) and pro-inflammatory cytokines; these rats also had altered oxidative stress status. In contrast, in rats treated with LP + MT (2 × 10(9) cfu/ml [∼ 2 mg/kg] + 0.5 mg MT/kg BW) for a total of 7 days before, concurrent with or after AFB1 treatment, there was a significant blockade/mitigation of each AFB1-impacted parameter. Moreover, treatment with the mixture at any point in relation to AFB1 treatment expectedly caused enhanced TNFα and IL-1ß expression relative to control values; all other parameters were comparable to values noted in control rats. Alone, the mixture had no impact on host parameters. From the results here it may be concluded the the LP + MT mixture was effective in protecting these hosts against AFB1-induced immunologic/physiologic disorders and that LP + MT could prevent and/or mitigate AFB1 toxicities in vivo.

Interaction of aflatoxin B1 and fumonisin B1 in mice causes immunotoxicity and oxidative stress: Possible protective role using lactic acid bacteria.

Aflatoxins (AF) are important foodborne mycotoxins implicated in human health and have immunocytotoxic effects. The aims of this study were to evaluate a new aflatoxin B1 (AFB1) and fumonisin B1 (FB1)-binding/degrading micro-organism for biological detoxification, to examine its ability to degrade AFB1 and FB1 in liquid medium, and to evaluate its potential in vivo protective role against any combined effects from AFB1 and FB1 on host splenocyte caspase-3 activity (reflecting DNA damage/cell death) and mRNA levels of select inflammation-regulating cytokines. Balb/c mice were divided into groups (10/group) and treated daily for 2 weeks by oral gavage with AFB1 (80 µg/kg BW), FB1 (100 µg/kg), AFB1 + FB1, or lactic acid bacteria (Lactobacillus paracasei BEJ01, 2 × 10(9) CFU/L, ∼2 mg/kg) - alone or in combination with the AFB1 and/or FB1. After the exposures, spleens were collected for measures of caspase-3 activity, lipid peroxidation (LP), and glutathione (GSH) content, expression of anti-oxidation protective enzymes (GPx and SOD), and mRNA levels of inflammation-regulating cytokines (e.g. IL-10, IL-4, IFNγ, TNFα). Thymii were also removed for analysis of apoptosis. The results indicated that, in the spleen, exposure to the mycotoxins led to increased caspase-3 activity, LP, and IL-10 and IL-4 mRNA levels, but decreased GSH content and down-regulated expression of GPx and SOD, and of IFNγ and TNFα mRNA. Co-treatment using Lactic Acid Bacteria (LAB) with AFB1 or FB1 suppressed levels of DNA fragmentation, normalized splenic LP and increased GSH levels, up-regulated expression of GPx and SOD, and normalized mRNA levels of the analyzed cytokines. It is concluded that AFB1 and FB1 might have combinational (synergistic moreso than additive) toxic effects in situ. Further, it can be seen that use of LAB induced protective effects against the oxidative stress and (immuno)toxicity of these agents in part through adhesion (and so likely diminished bioavalability).

Tunisian radish (Raphanus sativus) extract prevents cadmium-induced immunotoxic and biochemical alterations in rats.

Cadmium (Cd), a known carcinogen and potent immunotoxicant in humans and animals, is dispersed throughout the environment as a result of pollution from a variety of sources. Tunisian radish (Raphanus sativus) extract (TRE) is a known anti-oxidant and free radical scavenger that has been shown to help alleviate immune system disorders, including some induced by environmental toxicants. The present study was undertaken to investigate potential protective effects of TRE against Cd-induced immunotoxicities (and general toxicities) in situ. Cadmium chloride (at 2.5 mg CdCl2/kg BW) and TRE (5, 10, or 15 mg/kg BW) were given (alone or in combination [actually, in sequence of Cd and then TRE]) to rats daily by oral gavage for 2 weeks. Results indicated that treatment with CdCl2 alone resulted in significant decreases in plasma levels of total protein, triglycerides, creatine kinase, creatinine, IgG and IgA, T-lymphocyte sub-types (CD4(+), CD3(+), CD56(+), and CD8(+)), and in thymic and hepatic indices (relative weights). In contrast, CdCl2 treatment caused significant increases in serum LDH, AST, and ALT, in the formation/release of pro-inflammatory cytokines (IL-1 and TNFα), and in the relative weights of host spleen and kidneys. Rats treated with TRE alone had no discernable changes compared to the controls with regard to all test parameters. Combined treatment of CdCl2 and TRE-at any dose-resulted in a significant improvement of all test parameters compared to those seen with Cd alone. These results illustrated (and provided further support for a continuing belief in) the beneficial effects of TRE in reducing the harmful outcomes of commonly encountered toxicants (like Cd) on the immune system and on overall host health status.

Potential preventive role of lactic acid bacteria against aflatoxin M1 immunotoxicity and genotoxicity in mice.

Aflatoxin M1 (AFM1) is a mycotoxin produced by numerous Aspergillus species in pre- or post-harvest cereals and milk. Exposure to AFM1 imparts potent economic losses in the livestock industry. Toxicologically, it also causes severe immune system problems. The aims of this study were to evaluate a new AFM1-binding/degrading microorganism for biologic detoxification, to examine its ability to degrade AFM1 in liquid medium, and to evaluate its potential for in vivo preventative effects against AFM1-induced immunotoxicity and genotoxicity in mice. Lactobacillus plantarum MON03 (LP) isolated from Tunisian artisanal butter was found to display significant binding ability to AFM1 in PBS (93%) within 24 h of incubation. Further, the LP was able to tolerate gastric acidity, bile salts, and adhere efficiently to Caco-3 cells in vitro. The in vivo study used Balb/c mice that received either vehicle (control), LP only (at 1 × 10(9)CFU/L, ∼1 mg/kg bw), AFM1 (100 mg/kg bw), or AFM1 + LP daily for 15 days (by gavage); two other groups received a single dose of colchicine (4 mg/kg) or mitomycin C (1 mg/kg) as positive controls for induction of micronuclei and chromosomal aberrations, respectively. The results showed that, compared to in control mice, AFM1 treatment led to significantly decreased body weight gains, and caused cytotoxic/genotoxic effects as indicated by increases in frequencies of polychromatic erythrocytes, as well as those with micronucleation (PCEMN) and chromosomal aberrations, among bone marrow cells. The concurrent administration of LP with AFM1 strongly reduced the adverse effects of AFM1 on each parameter. Mice receiving AFM1 + LP co-treatment displayed no significant differences in the assayed parameters as compared to the control mice. By itself, the bacteria caused no adverse effects. Based on the data, it is concluded that the test bacteria could potentially be beneficial in the detoxification of AFM1-contaminated foods and feeds for humans and animals.

Lactobacillus paracasei BEJ01 prevents immunotoxic effects during chronic zearalenone exposure in Balb/c mice.

BACKGROUND AND AIM: Zearalenone (ZEN) is an estrogenic mycotoxin produced by numerous Fusarium species in pre- or post-harvest cereals. ZEN displays a potent estrogenicity in livestock and also causes severe immunological problems. The aims of this study were to isolate a new ZEN-degrading micro-organism for biological detoxification, to examine its ability to degrade ZEN in liquid medium, and to evaluate its potential for in vivo preventitive effects against ZEN (as would occur with contaminated feed)-induced immunomodulation in mice. MATERIALS AND METHODS: Lactobacillus paracasei BEJ01 (LP) isolated from Tunisian artisanal butter was found to display significant binding ability to ZEN in phosphate-buffered saline (i.e. 96.6%) within 24 h of incubation. The in vivo study was conducted using Balb/c mice that received either vehicle (control), LP only (at 2 × 10(9 )cfu/l, ∼2 mg/kg BW), ZEN alone (at 40 mg/kg BW), or ZEN + LP daily for 15 d. RESULTS: Compared to control mice, ZEN treatment led to significantly decreased body weight gains and decrements in all immune parameters assessed. The addition of LP to ZEN strongly reduced the adverse effects of ZEN on each parameter. In fact, mice receiving ZEN + LP co-treatment displayed no significant differences in the assayed parameters as compared to the control mice. The exposures to the bacteria alone had no adverse effects in the mice. CONCLUSION: From these data, we conclude that LP bacteria could be beneficial in human and animals for protection against immunotoxicity from ZEN at high levels and during chronic exposures.

Interaction of Lactobacillus plantarum MON03 with Tunisian montmorillonite clay and ability of the composite to immobilize Zearalenone in vitro and counteract immunotoxicity in vivo.

BACKGROUND AND AIM: The present study was conducted to determine the abilities of the living Lactobacillus plantarum MON03 (LP) cells, Tunisian montmorillonite clay and their composites to accumulate Zearalenone (ZEA) from a liquid medium and elucidate the preventive effect of their composite in ZEA-contaminated balb/c mice showing immunotoxicity disorders. MATERIALS AND METHODS: In the in vitro study, LP (2 × 10(9) CFU/mL), TM (0.5 mg) and LP+TM were incubated with 50 µg mL(-1) ZEA for 0, 12 and 24 h. For the in vivo study, the composite MT+LP was evaluated also for possible protection regarding ZEA-immunotoxicity in Balb/c mice as a sensitive model. RESULTS: Results indicated that TM and LP+TM had a high capacity of adsorbing ZEA 87.2 ± 2.1 and 94.2 ± 2.1%, respectively. However, LP alone able to remove only 78% after 24 h of incubation. The quantity of adsorbed ZEA by LP, TM and LP+TM were 39, 43,5 and 47 µg mL(-1) of PBS, respectively. The in vivo results indicated that mice orally exposed to ZEA- (40 mg/kg bw) for 2 weeks showed severe immunotoxicity typical of fusarotoxicosis regarding thymocytes and splenocytes cell viability count, IFN-γ, IL-12, TNF-α production and B-cell activation. Mice treated with LP and TM alone, and LP+MT in combination with ZEA were comparable to the control. CONCLUSION: Both LP and TM are safe by themselves and their composite succeeded to exert a potential prevention by counteracting ZEA-immunotoxicity and can be implicated in the biotechnology of ZEA removal in human food and animal feed.