Processes influencing the toxicity of microplastics ingested through the diet.

The present study assesses the effect of culinary treatment and gastrointestinal digestion upon the release of additives present in microplastics. Organic additives were determined by gas chromatography-mass spectrometry, and inorganic additives using inductively coupled plasma-mass spectrometry. The results revealed a large number of organic additives in the plastic samples, some being classified as possible carcinogens. Contents of Sb in PET (polyethylene terephthalate), Zn and Ba in LDPE (low-density polyethylene) and PVC (polyvinylchloride), and Ti and Pb in LDPE were also noteworthy. The culinary process promotes the release and solubilization of additives into the cooking liquid, with phthalates, benzophenone, N-butylbenzenesulfonamide (NBBS) and bisphenol A being of particular concern. The solubilization of phthalates and NBBS was also observed during gastrointestinal digestion. This study demonstrates that culinary treatment and gastrointestinal digestion promote release and solubilization of additives from plastics ingested with the diet. Such solubilization may facilitate their entry into the systemic circulation.

Phosphate Uptake and Its Relation to Arsenic Toxicity in Lactobacilli.

The use of probiotic lactobacilli has been proposed as a strategy to mitigate damage associated with exposure to toxic metals. Their protective effect against cationic metal ions, such as those of mercury or lead, is believed to stem from their chelating and accumulating potential. However, their retention of anionic toxic metalloids, such as inorganic arsenic, is generally low. Through the construction of mutants in phosphate transporter genes (pst) in Lactiplantibacillus plantarum and Lacticaseibacillus paracasei strains, coupled with arsenate [As(V)] uptake and toxicity assays, we determined that the incorporation of As(V), which structurally resembles phosphate, is likely facilitated by phosphate transporters. Surprisingly, inactivation in Lc. paracasei of PhoP, the transcriptional regulator of the two-component system PhoPR, a signal transducer involved in phosphate sensing, led to an increased resistance to arsenite [As(III)]. In comparison to the wild type, the phoP strain exhibited no differences in the ability to retain As(III), and there were no observed changes in the oxidation of As(III) to the less toxic As(V). These results reinforce the idea that specific transport, and not unspecific cell retention, plays a role in As(V) biosorption by lactobacilli, while they reveal an unexpected phenotype for the lack of the pleiotropic regulator PhoP.

Protective effects of oral administration of lactic acid bacteria strains against methylmercury-induced intestinal toxicity in a murine model.

The utilization of lactic acid bacteria has been proposed to mitigate the burden of heavy metal exposure through processes probably involving chelation and reduced metal bioaccessibility. We evaluated the effects of daily intake of two strains of lactobacilli (Lactobacillus intestinalis LE1 or Lactobacillus johnsonii LE2) on intestinal toxicity during methylmercury (MeHg) exposure through drinking water (5 mg/L) for two months in mice. MeHg exposure resulted in inflammation and oxidative stress at the colon, as well as an increase in intestinal permeability accompanied by decreased fecal short-chain fatty acids (SCFA). The administration of the strains resulted in a differential protective effect that, based on their chelation capacity, supported the existence of additional mechanisms of action besides chelation. Both strains reduced IL-1ß levels and oxidative stress, while LE1 lowered TNF-α, diminished MeHg-induced mucus over-secretion triggered by the IL-4/IL-13/STAT6 pathway, reduced intestinal permeability, and ameliorated inflammation and oxidative stress, probably by acting on the Keap1/Nrf2/ARE pathway. Administration of LE1 partially restored SCFA contents, which could be partly responsible for the positive effects of this strain in alleviating MeHg toxicity. These results demonstrate that lactobacilli strains can be useful tools in reducing the intestinal toxicity of MeHg, the main mercurial form conveyed by food.

Impact of Chronic Exposure to Arsenate through Drinking Water on the Intestinal Barrier.

Chronic exposure to inorganic arsenic (As) [As(III) + As(V)], which affects millions of people, increases the incidence of some kinds of cancer and other noncarcinogenic pathologies. Although the oral pathway is the main source of exposure, in vivo studies conducted to verify the intestinal toxicity of this metalloid are scarce and are mainly focused on evaluating the toxicity of As(III). The aim of this study was to evaluate the effect of chronic exposure (6 months) of BALB/c mice to As(V) (15-60 mg/L) via drinking water on the different components of the intestinal barrier and to determine the possible mechanisms involved. The results show that chronic exposure to As(V) generates a situation of oxidative stress (increased lipid peroxidation and reactive species) and inflammation (increased contents of several proinflammatory cytokines and neutrophil infiltrations) in the intestinal tissues. There is also evidence of an altered expression of constituent proteins of the intercellular junctions (Cldn1, Cldn3, and Ocln) and the mucus layer (Muc2) and changes in the composition of the gut microbiota and the metabolism of short-chain fatty acids. All of these toxic effects eventually may lead to the disruption of the intestinal barrier, which shows an increased paracellular permeability. Moreover, signs of endotoxemia are observed in the serum of As(V)-treated animals (increases in lipopolysaccharide-binding protein LBP and the proinflammatory cytokine IL-1ß). The data obtained suggest that chronic exposure to As(V) via drinking water affects the intestinal environment.

Challenges and strategies for preventing intestinal damage associated to mercury dietary exposure.

Food represents the major risk factor for exposure to mercury in most human populations. Therefore, passage through the gastrointestinal tract plays a fundamental role in its entry into the organism. Despite the intense research carried out on the toxicity of Hg, the effects at the intestinal level have received increased attention only recently. In this review we first provide a critical appraisal of the recent advances on the toxic effects of Hg at the intestinal epithelium. Next, dietary strategies aimed to diminish Hg bioavailability or modulate the epithelial and microbiota responses will be revised. Food components and additives, including probiotics, will be considered. Finally, limitations of current approaches to tackle this problem and future lines of research will be discussed.

Oral exposure to inorganic mercury or methylmercury elicits distinct pro-inflammatory and pro-oxidant intestinal responses in a mouse model system.

Humans are mainly exposed to mercury (Hg) through contaminated foodstuffs. However, the effects of Hg on the intestinal tract have received little attention. We performed a subchronic exposure to inorganic mercury or methylmercury in mice through drinking water (1, 5 or 10 mg/L for four months) to evaluate their intestinal impact. Histological, biochemical and gene expression analyses showed that both Hg species induced oxidative stress in small intestine and colon, while inflammation was mainly detected in the colon. Increased fecal albumin content indicated a compromised epithelial barrier. Mucus production was possibly also affected, as an increase in Muc2 expression was detected. However, differential effects were detected between both Hg species. Activation of p38 MAPK and increased crypt depth were detected in colon only with MeHg. Minor differences in microbiota composition were detected between unexposed and exposed mice. Although significant differences were detected between both Hg species at 10 mg/L, only the relative abundances of low abundance taxa were affected. Concentrations of microbial-derived short-chain fatty acids were decreased, suggesting an effect on microbial metabolism or increased demand by the intestinal epithelium. Results obtained confirm previous in vitro studies and highlights the intestinal mucosa as an initial target of Hg.

Lactic acid bacteria strains reduce in vitro mercury toxicity on the intestinal mucosa.

A bicameral model consisting of Caco-2 and HT29-MTX intestinal epithelial cells and THP-1-derived macrophages has been used to test the ability of two strains of Lactobacillus to protect from damage caused by mercury. Exposure to 1 mg/ml mercury [Hg(II) or methyl-Hg] for seven days in this model resulted in an inflammatory and pro-oxidant response mainly driven by macrophages. This led to an impairment in the intestinal barrier, defective tight-junctions, increased permeability and mucus hypersecretion. In addition, the wound-healing capacity of the epithelial monolayer was also diminished. However, the presence of heat-killed Lactobacillus intestinalis or Lactobacillus johnsonii cells during Hg exposure reverted these effects, and most of the parameters recovered values similar to control cells. Both lactobacilli showed the capacity to bind Hg(II) and methyl-Hg under the cell culture conditions. This points to Hg sequestration as a likely mechanism that counteracted Hg toxicity. However, differences in the Hg binding capacity and in the effects between both strains suggest that other probiotic-mediated mechanisms may play a role in the alleviation of the damage elicited by Hg. These results show the potential of the bicameral intestinal epithelial model for screening of effective strains for their use in later in vivo studies.

Mercury toxic effects on the intestinal mucosa assayed on a bicameral in vitro model: Possible role of inflammatory response and oxidative stress.

Exposure to mercury (Hg) mostly occurs through diet, where it is mainly found as inorganic Hg [Hg(II)] or methylmercury (MeHg). In vivo studies have linked its exposure with neurological and renal diseases, however, its toxic effects upon the gastrointestinal tract are largely unknown. In order to evaluate the effect of Hg on intestinal mucosa, a bicameral system was employed with co-cultures of Caco-2 and HT29-MTX intestinal epithelial cells and THP-1 macrophages. Cells were exposed to Hg(II) and MeHg (0.1, 0.5, 1 mg/L) during 11 days. The results evidenced a greater pro-inflammatory response in cells exposed to Hg with increments of IL-8 (15-126%) and IL-1ß release (39-63%), mainly induced by macrophages which switched to a M1 phenotype. A pro-oxidant response was also observed in both cell types with an increase in ROS/RNS levels (44-140%) and stress proteins expression. Intestinal cells treated with Hg displayed structural abnormalities, hypersecretion of mucus and defective tight junctions. An increased paracellular permeability (123-170%) at the highest concentrations of Hg(II) and MeHg and decreased capacity to restore injuries in the cell monolayer were also observed. All these toxic effects were governed by various inflammatory signalling pathways (p38 MAPK, JNK and NF-κB).

Dietary microplastics: Occurrence, exposure and health implications.

The increasing use of plastic materials generates an enormous amount of waste. In the aquatic environment, a significant part of this waste is present in the form of microplastics (MPs)- particles with a diameter of between 0.1 µm and 5 mm. The arrival of these small plastics in the food chain has been recently documented. MPs have been reported in fishery products, drinking water and sea salt among other foods. Their intestinal absorption is considered limited due to their size, however, they contain a mixture of chemicals intentionally added during their manufacture, which could cross the intestinal barrier. Currently there are not enough data to allow an accurate assessment of the risk associated with dietary exposure to MPs. The lack of robust methodologies is undoubtedly one of the main problems. There is limited information on occurrence in dietary sources (drinking water and food), human intake, toxicokinetics and long term toxicity of these contaminants. The present review describes the studies published so far and points to the need for improved knowledge in order to have a more accurate view of the problems posed by MPs.

Arsenic in Tissues and Prey Species of the Scalloped Hammerhead (Sphyrna lewini) from the SE Gulf of California.

The bioaccumulation of arsenic (As) in the muscle, liver, kidneys, and brain of the shark Sphyrna lewini was measured in 40 juvenile specimens from southeast Gulf of California. Additionally, the biomagnification factor was calculated through prey items from stomach contents of the analyzed specimens. The concentrations of As (mg kg-1, wet weight) were higher in the muscle (10.1 ± 0.3) and liver (9.4 ± 0.5) than in the brain (4.5 ± 0.3) and kidneys (4.2 ± 0.2), which may be attributed to the biological functions of each tissue. Positive correlations were found between the levels of As in muscle and liver with the biological parameters of S. lewini. Hammerhead sharks feed mainly of teleost fishes with low As values (Clupeidae fishes, 1.1 ± 0.5; Sciaenidae fishes, 1.0 ± 0.6; Scomber japonicus, 1.2 ± 0.6; and Etropus crossotus 2.1 ± 0.4) compared with the predator, indicating biomagnification. Inorganic arsenic (Asi) in muscle was estimated as 3% of the total As, although muscle consumption is unlikely to represent a risk (HQ < 1) in humans. Moreover, the probabilities of developing cancer were estimated as low (3.99 × 10-5 to 3.32 × 10-6). To avoid health risks related to As, a weekly ration must not exceed 69.3 and 484.8 g in children and adults, respectively.

Arsenic speciation in cooked food and its bioaccessible fraction using X-ray absorption spectroscopy.

Thermal processing or the digestion process can alter the forms of arsenic (As) present in food. Identification of As species is necessary to accurately determine the risk associated with food consumption. X-ray absorption near-edge structure (XANES) was used to investigate As species in rice, asparagus, and garlic boiled in water containing As(V), and in their bioaccessible fractions (solubilized As after gastrointestinal digestion). The XANES analysis revealed the presence of As(III) (11871.5 eV) or As(III)-S [As(III)-Cys, 11869.6 eV] solution in the cooked foods and in their bioaccessible fractions. The percentage of trivalent species (12-55%) followed the order asparagus ≫ rice ≈ garlic. In the asparagus and garlic samples, part of the As(V) (tetrahedral form) [11875 eV] that had been added appeared in the form of an octahedral As(V) compound [As(V)-glycerol, 11876 eV]. All these changes could considerably modify the risk associated with ingestion of As-contaminated food.

In Vitro Evaluation of the Protective Role of Lactobacillus StrainsAgainst Inorganic Arsenic Toxicity.

Inorganic arsenic [iAs, As(III) + As(V)] is considered a human carcinogen. Recent studies show that it has also toxic effects on the intestinal epithelium which might partly explain its systemic toxicity. The aim of this study is to evaluate the protective role of lactic acid bacteria (LAB) against the toxic effects of iAs on the intestinal epithelium. For this purpose, the human colonic cells Caco-2 were exposed to As(III) in the presence of various LAB strains or their conditioned medium. Results showed that some strains and their conditioned media partially revert the oxidative stress, the production of pro-inflammatory cytokines, the alterations of the distribution of tight junction proteins, and the cell permeability increases caused by As(III). These results show that both soluble factors secreted or resulting from LAB metabolism and cell-cell interactions are possibly involved in the beneficial effects. Therefore, some LAB strains have potential as protective agents against iAs intestinal barrier disruption.

Toxic trace elements in dried mushrooms: Effects of cooking and gastrointestinal digestion on food safety.

Mushrooms can accumulate toxic trace elements. The objectives of the present study are to evaluate levels of mercury, cadmium, lead, and arsenic in dried mushrooms, to determine the effect of cooking on the contents of these elements, and to evaluate their bioaccessibility in the mushrooms ready for consumption. The results showed that Hg levels in Amanita ponderosa, Boletus edulis, Marasmius oreades, and Tricholoma georgii, as well as Cd levels in some samples of Amanita caesarea and T. georgii, exceeded the legislated limits. Cooking significantly reduced the levels of As (26-72%), whereas the reduction in levels of Hg, Cd, and Pb was much lower. However, the bioaccessibility of As (63-81%) was higher than the values obtained for the metals (<40%). Taking the effects of cooking and gastrointestinal digestion into account gives a more realistic estimate of the risk associated with the consumption of mushrooms.

Inorganic arsenic causes intestinal barrier disruption.

Inorganic arsenic (As) is the most toxic form of As found in food and water. Gastrointestinal disorders have been reported in populations chronically exposed to this arsenical form or to one of its metabolites; however, studies to determine the mechanisms of inorganic As toxicity at the intestinal level are scarce. The aim of this study is to determine the mechanisms of toxicity of inorganic As [As(iii) and As(v)] on intestinal epithelial cells. For this purpose, two human intestinal cell models were used: non-transformed colon epithelial cells (NCM460) and epithelial cells from a colorectal adenocarcinoma (Caco-2). Exposure to As(iii) and As(v) generates an increase in the release of the pro-inflammatory cytokine IL-8 (57-1135%) and an increase in the generation of reactive oxygen and/or nitrogen species (130-340%) in both cell lines. This pro-inflammatory and pro-oxidant response may be responsible for the structural and functional modifications demonstrated in the monolayers formed by both cell types. Treatments with As(iii) and As(v) produce a redistribution of zonula occludens 1 and a reduction in the expression of claudin 1, tight junction proteins that participate in maintaining the structure of the epithelium. All these toxic effects are finally translated into a loss of the barrier function of intestinal monolayers.

In vivo evaluation of the effect of arsenite on the intestinal epithelium and associated microbiota in mice.

Chronic exposure to inorganic arsenic (As) [As(III) + As(V)], which affects millions of people, increases the incidence of some kinds of cancer and other non-carcinogenic pathologies. Although the oral pathway is the main form of exposure, in vivo studies have not been conducted to verify the intestinal toxicity of this metalloid. The aim of this study is to perform an in vivo evaluation of the intestinal toxicity of inorganic As, using female BALB/c mice exposed through drinking water to various concentrations of As(III) (20, 50, and 80 mg/L) for 2 months. An increase was observed in oxygen and/or nitrogen reactive species, and in gene and protein expression of pro-inflammatory cytokines (IL-1ß, IL-2, IL-6) at concentrations equal to or greater than 50 mg/L. These changes were accompanied by a profound remodeling of the intestinal microbial profile in terms of diversity and global composition, which could be at the basis or exacerbate As(III) toxic effects. The histological study showed that there was moderate inflammation of the mucosa and submucosa, accompanied by hyperplasia of crypts at the highest administered dose. In addition, all the treatments with As(III) resulted in a decreased expression of Muc2, which encodes one of the main components of the intestinal layer of mucus. The effects described are compatible with the increased intestinal permeability observed at concentrations equal to or greater than 50 mg/L, indicative of loss of barrier function.

Dietary Compounds To Reduce In Vivo Inorganic Arsenic Bioavailability.

It is estimated that approximately 200 million people are exposed to arsenic levels above the World Health Organization provisional guideline value, and various agencies have indicated the need to reduce this exposure. In view of the difficulty of removing arsenic from water and food, one alternative is to reduce its bioavailability (the amount that reaches the systemic circulation after ingestion). In this study, dietary components [glutathione, tannic acid, and Fe(III)] were used to achieve this goal. As(III) or As(V) (1 mg/kg body weight) was administered daily to BALB/c mice, along with the dietary components, for 15 days. The results confirm the efficacy of Fe(III) and glutathione as reducers of arsenic bioavailability and tissue accumulation. Also, these treatments did not result in reductions of Ca, K, P, and Fe contents in the liver. These data suggest that use of these two compounds could be part of valid strategies for reducing inorganic arsenic exposure in chronically exposed populations.

Effect of lactic acid bacteria on mercury toxicokinetics.

The capacity of two LAB strains to inhibit inorganic [Hg(II)] and organic (methyl-Hg; MeHg) mercury translocation through monolayers of co-cultures of NCM460 and HT29-MTX colonic cells was evaluated. Lactobacillus casei BL23 and Lactobacillus acidophilus ATCC4356 reduced the permeability of Hg(II) and MeHg from aqueous solutions through NCM460/HT29-MTX monolayers (20-94% reduction). However, assays using the bioaccessible (soluble) Hg fraction obtained by in vitro gastrointestinal digestion of Hg-contaminated swordfish only showed a reduction (42%) with the BL23 strain. In vivo experiments carried out in mice receiving an acute dose of Hg(II) or MeHg (0.5 mg/kg body weight/day) with or without lactobacilli resulted in significant decreases of the bioavailability of MeHg with both strains and increased excretion of Hg in feces after treatment with the lactobacilli. However, Hg(II) bioavailability or excretion was not affected. Hg accumulation in liver and kidney remained similar in LAB-treated or non-treated animals. This is the first study of the impact of LAB on Hg(II) and MeHg toxicokinetics and shows that some LAB strains have potential to diminish MeHg bioavailability. Furthermore, it has established the basis for new studies on the protective effect of LAB under conditions resembling subchronic and chronic Hg exposures.

Arsenic exposure of child populations in Northern Argentina.

Chronic exposure to inorganic arsenic (As) is associated with numerous adverse effects. Argentina is one of the countries affected by arsenicism; however, there are few studies that evaluate inorganic As exposure and its effects on child population. The aim of this study is to evaluate exposure to As through water and food in child populations living in the provinces of Santiago del Estero and Chaco (n = 101), and to determine the impact of this exposure analysing biomarkers of exposure (urine and hair As contents) and effect [8-oxo-7,8-dihydro-2'-deoxyguanosine (8-OHdG)]. The populations selected live in three areas with different levels of As in the drinking water (Santa Teresa de Carballo, 0.925 mg/L; Taco Pozo, 0.210 mg/L; Jumi Pozo, 0.016 mg/L). The As intakes through water and food are especially high in the areas with the greatest As exposure (Santa Teresa de Carballo, 1575 ±â€¯8 µg/day; Taco Pozo, 386 ±â€¯8 µg/day; Jumi Pozo, 39 ±â€¯1 µg/day). The total As contents in most of the samples of hair (0.11-13.11 mg/kg) and urine (31-4258 µg/g creatinine) are higher than the reference values (hair: 1 mg/kg; urine: 50 µg/g creatinine). The increase in the level of As exposure alters the profile of metabolites in urine, with a decrease of dimethylarsinic acid (10%) and an increase in the percentages of monomethylarsonic acid (4%) and inorganic As (6%). The results also show high values of 8-OHdG (3.7-37.8 µg/g creatinine), a oxidative DNA damage marker, in the two areas with greater As exposure.

Effect of chronic exposure to inorganic arsenic on intestinal cells.

Chronic exposure to inorganic arsenic (As)-As(III) + As(V)-is associated with type 2 diabetes, vascular diseases and various types of cancer. Although the oral route is the main way of exposure to inorganic As, the adverse gastrointestinal effects produced by chronic exposure are not well documented. The aim of the present study is to evaluate the effect of chronic exposure to As(III) on the intestinal epithelium. For this purpose, NCM460 cells, non-transformed epithelial cells from the human colon, were exposed to As(III) (0.01-0.2 mg/L) for 6 months and monitored for acquisition of a tumor-like phenotype. Secretion of matrix metalloproteinases, histone modifications (H3 acetylation), hyperproliferation capacity, formation of floating spheres, anchorage-independent growth, release of cytokine interleukin-8 and expression of relevant genes in colon tumorigenesis were assessed. The results show a maintained proinflammatory response from the beginning, with an increase in interleukin-8 secretion (≤570%). Downregulation of CDX1 and CDX2 was also observed. After 14 weeks of exposure, cells presented marked increases in matrix metalloproteinase-2 secretion and histone modifications. As(III)-treated cells were hyperproliferative, grew in low-serum media and were able to form free-floating spheres. Overall, these data suggest that exposure of human colon epithelial cells to As(III) facilitates acquisition of transformed cell characteristics.

Use of lactic acid bacteria and yeasts to reduce exposure to chemical food contaminants and toxicity.

Chemical contaminants that are present in food pose a health problem and their levels are controlled by national and international food safety organizations. Despite increasing regulation, foods that exceed legal limits reach the market. In Europe, the number of notifications of chemical contamination due to pesticide residues, mycotoxins and metals is particularly high. Moreover, in many parts of the world, drinking water contains high levels of chemical contaminants owing to geogenic or anthropogenic causes. Elimination of chemical contaminants from water and especially from food is quite complex. Drastic treatments are usually required, which can modify the food matrix or involve changes in the forms of cultivation and production of the food products. These modifications often make these treatments unfeasible. In recent years, efforts have been made to develop strategies based on the use of components of natural origin to reduce the quantity of contaminants in foods and drinking water, and to reduce the quantity that reaches the bloodstream after ingestion, and thus, their toxicity. This review provides a summary of the existing literature on strategies based on the use of lactic acid bacteria or yeasts belonging to the genus Saccharomyces that are employed in food industry or for dietary purposes.