Effect of topical berberine in murine cutaneous leishmaniasis lesions.

OBJECTIVES: More effective topical treatments remain an unmet need for the localized forms of cutaneous leishmaniasis (CL). The aim of this study was to evaluate the efficacy and safety of a topical berberine cream in BALB/c mice infected with Leishmania major parasites. METHODS: A cream containing 0.5% berberine-ß-glycerophosphate salt and 2.5% menthol was prepared. Its physicochemical and stability properties were determined. The cream was evaluated for its capacity to reduce lesion size and parasitic load as well as to promote wound healing after twice-a-day administration for 35 days. Clinical biochemical profile was used for estimating off-target effects. In vitro time-to-kill curves in L. major-infected macrophages and skin and plasma pharmacokinetics were determined, aiming to establish pharmacokinetic/pharmacodynamic relationships. RESULTS: The cream was stable at 40°C for 3 months and at 4°C for at least 8 months. It was able to halt lesion progression in all treated mice. At the end of treatment, parasite load in the skin was reduced by 99.9% (4 log) and genes involved in the wound healing process were up-regulated compared with untreated mice.The observed effects were higher than expected from in vitro time-to-kill kinetic and plasma berberine concentrations, which ranged between 0.07 and 0.22 µM. CONCLUSIONS: The twice-a-day administration of a topical berberine cream was safe, able to stop parasite progression and improved the appearance of skin CL lesions. The relationship between drug plasma levels and in vivo effect was unclear.

Selection of Enzymatic Treatments for Upcycling Lentil Hulls into Ingredients Rich in Oligosaccharides and Free Phenolics.

In this study, the comprehensive chemical characterization of red lentil hulls obtained from the industrial production of football and split lentils was described. The lentil hulls were rich in dietary fiber (78.43 g/100 g dry weight with an insoluble to soluble fiber ratio of 4:1) and polyphenols (49.3 mg GAE/g dry weight, of which 55% was bound phenolics), which revealed the suitability of this lentil by-product as a source of bioactive compounds with recognized antioxidant and prebiotic properties. The release of oligosaccharides and phenolic compounds was accomplished by enzymatic hydrolysis, microwave treatment and a combination of both technologies. The key role played by the selection of a suitable enzymatic preparation was highlighted to maximize the yield of bioactive compounds and the functional properties of the lentil hull hydrolysates. Out of seven commercial preparations, the one with the most potential for use in a commercial context was Pectinex® Ultra Tropical, which produced the highest yields of oligosaccharides (14 g/100 g lentil hull weight) and free phenolics (45.5 mg GAE/100 g lentil hull weight) and delivered a four-fold increase in terms of the original antioxidant activity. Finally, this enzyme was selected to analyze the effect of a microwave-assisted extraction pretreatment on the yield of enzymatic hydrolysis and the content of free phenolic compounds and oligosaccharides. The integrated microwave and enzymatic hydrolysis method, although it increased the solubilization yield of the lentil hulls (from 25% to 34%), it slightly decreased the content of oligosaccharides and proanthocyanidins and reduced the antioxidant activity. Therefore, the enzymatic hydrolysis treatment alone was more suitable for producing a lentil hull hydrolysate enriched in potential prebiotics and antioxidant compounds.

Potential Usefulness of a Wakame/Carob Functional Snack for the Treatment of Several Aspects of Metabolic Syndrome: From In Vitro to In Vivo Studies.

Metabolic syndrome (MetS) greatly increases the risk of cardiovascular diseases and type 2 diabetes mellitus. The aim of this study was to evaluate the efficacy of functional snacks containing a combination of wakame (W) and carob pod (CP) flours in reducing markers associated with MetS. The mechanisms of action underlying these effects were also evaluated. In vitro approaches were carried out in mature 3T3-L1 adipocytes and RAW 264.7 macrophages treated with different doses of extracts from W, CP, or a combination of both. Furthermore, an in vivo experiment was conducted in rats with MetS treated with normal-caloric diets containing different snack formulations with combinations of 1/50 (snack A) or 1/5 of wakame/carob (snack B). In vitro experiments results indicated that both W and CP had delipidating effects, but only the latter induced anti-inflammatory and anti-hypertensive effects. As far as the in vivo study is concerned, snack B was ineffective and snack A showed an anti-hypertensive effect in rats with MetS. The present study shows for the first time the in vitro efficacy of a W and CP combination as an anti-inflammatory, delipidating, and anti-hypertensive tool, and its potential usefulness in treating MetS.

Short communication: Analysis of mycotoxins in Spanish milk.

We surveyed the presence of 22 mycotoxins in 191 Spanish cow milk samples. Mycotoxins could be carried over from diet into animal milk and have toxic effects on human and animal health. The interaction of different mycotoxins may be additive or synergetic. Therefore, surveillance of mycotoxin co-occurrence in milk is recommended. Aflatoxins M1, B1, B2, G1, and G2, ochratoxins A and B, nivalenol, deoxynivalenol, deepoxy-deoxynivalenol, 3- and 15-acetyldeoxynivalenol, diacetoxyscirpenol, neosolaniol, fusarenon X, T-2 and HT-2 toxins, fumonisins B1, B2, and B3, sterigmatocystin, and zearalenone were analyzed. Samples were treated by liquid-liquid extraction with acidified acetonitrile, followed by an acetonitrile-water phase separation using sodium acetate. The analysis was carried out by HPLC coupled to a triple quadrupole mass spectrometer. None of the analyzed mycotoxins had a concentration level higher than their detection limit (0.05-10.1 µg/L). The aflatoxin M1 in the samples never exceeded the level established by the European Union.

Differential Gene Expression by Lactobacillus plantarum WCFS1 in Response to Phenolic Compounds Reveals New Genes Involved in Tannin Degradation.

Lactobacillus plantarum is a lactic acid bacterium that can degrade food tannins by the successive action of tannase and gallate decarboxylase enzymes. In the L. plantarum genome, the gene encoding the catalytic subunit of gallate decarboxylase (lpdC, or lp_2945) is only 6.5 kb distant from the gene encoding inducible tannase (L. plantarumtanB [tanBLp ], or lp_2956). This genomic context suggests concomitant activity and regulation of both enzymatic activities. Reverse transcription analysis revealed that subunits B (lpdB, or lp_0271) and D (lpdD, or lp_0272) of the gallate decarboxylase are cotranscribed, whereas subunit C (lpdC, or lp_2945) is cotranscribed with a gene encoding a transport protein (gacP, or lp_2943). In contrast, the tannase gene is transcribed as a monocistronic mRNA. Investigation of knockout mutations of genes located in this chromosomal region indicated that only mutants of the gallate decarboxylase (subunits B and C), tannase, GacP transport protein, and TanR transcriptional regulator (lp_2942) genes exhibited altered tannin metabolism. The expression profile of genes involved in tannin metabolism was also analyzed in these mutants in the presence of methyl gallate and gallic acid. It is noteworthy that inactivation of tanR suppresses the induction of all genes overexpressed in the presence of methyl gallate and gallic acid. This transcriptional regulator was also induced in the presence of other phenolic compounds, such as kaempferol and myricetin. This study complements the catalog of L. plantarum expression profiles responsive to phenolic compounds, which enable this bacterium to adapt to a plant food environment.IMPORTANCELactobacillus plantarum is a bacterial species frequently found in the fermentation of vegetables when tannins are present. L. plantarum strains degrade tannins to the less-toxic pyrogallol by the successive action of tannase and gallate decarboxylase enzymes. The genes encoding these enzymes are located close to each other in the chromosome, suggesting concomitant regulation. Proteins involved in tannin metabolism and regulation, such GacP (gallic acid permease) and TanR (tannin transcriptional regulator), were identified by differential gene expression in knockout mutants with mutations in genes from this region. This study provides insights into the highly coordinated mechanisms that enable L. plantarum to adapt to plant food fermentations.

Antitumoural Sulphur and Selenium Heteroaryl Compounds: Thermal Characterization and Stability Evaluation.

The physicochemical properties of a compound play a crucial role in the cancer development process. In this context, polymorphism can become an important obstacle for the pharmaceutical industry because it frequently leads to the loss of therapeutic effectiveness of some drugs. Stability under manufacturing conditions is also critical to ensure no undesired degradations or transformations occur. In this study, the thermal behaviour of 40 derivatives of a series of sulphur and selenium heteroaryl compounds with potential antitumoural activity were studied. In addition, the most promising cytotoxic derivatives were analysed by a combination of differential scanning calorimetry, X-ray diffraction and thermogravimetric techniques in order to investigate their polymorphism and thermal stability. Moreover, stability under acid, alkaline and oxidative media was tested. Degradation under stress conditions as well as the presence of polymorphism was found for the compounds VA6E and VA7J, which might present a hurdle to carrying on with formulation. On the contrary, these obstacles were not found for derivative VA4J.

Molecular characterization of allergens in raw and processed kiwifruit.

BACKGROUND: The prevalence of allergy to kiwifruit is increasing in Europe since the last two decades. Different proteins have been identified as kiwifruit allergens; even though with geographic differences, Act d 1, a cysteine protease protein of 30 kDa, and Act d 2, a thaumatin-like protein of 24 kDa, are normally considered the most important. The aim of this study was (i) to identify at molecular level the sensitization pattern in a group of well-characterized patients allergic to kiwifruit and (ii) to assess the role of technological treatments on kiwifruit allergenic potential. METHODS: The differences in the pattern of antigenicity between fresh and processed kiwifruit were evaluated by both immunoelectrophoretic techniques and clinical tests. RESULTS: In the group of patients included in this study, three proteins were identified as major allergens in fresh kiwifruit, as the specific sensitization was present in ≥50% of the subjects. These proteins corresponded to actinidin (Act d 1), pectin methyl aldolase (Act d 6), and thaumatin-like protein (Act d 2). Kiwellin (Act d 5) and proteins of Bet v 1 family (Act d 8/act d 11) were also recognized as minor allergens. Immunoreactivity was totally eliminated by industrial treatments used for the production of kiwifruit strained derivative. CONCLUSIONS: In this group of allergic children, the technological treatments used in the production of kiwifruit strained product reduced drastically the allergenic potential of kiwifruit.

Allergenic proteins in enology: a review on technological applications and safety aspects.

Proteinaceous products are widely used as fining agents during winemaking to remove unwanted insoluble particles and undissolved microscopic particles (colloidal material) from the must or wine to improve stability. Some of them (egg white, caseinates, and fish gelatine) have allergenic potential and the presence of their residues in the final product could represent a risk for allergic individuals. Moreover, lysozyme (an egg allergen) is included among wine additives to control the fermentation processes and avoid spoiling during winemaking. The aim of this paper is to review the experimental/clinical data on the use of allergenic products in enology and the measurement of relative risk for sensitized subjects. In addition, methods developed specifically for the quantification of allergenic residues in must and wine are described.

Impact of Elicitation on Antioxidant and Potential Antihypertensive Properties of Lentil Sprouts.

The aim of this study was to investigate the application of elicitors (500 µM ascorbic acid, 50 µM folic acid, 5 mM glutamic acid and 50 ppm chitosan in 5 mM glutamic acid) during lentil germination up to 8 days as a strategy to increase germination rate and to enhance the accumulation of γ-aminobutyric acid (GABA) and phenolic compounds. The effect of elicitation on the protein profile and antioxidant and angiotensin I converting enzyme (ACE) inhibitory activities of sprouted lentils was also evaluated. The application of elicitors did not negatively affect the germination yield of lentils and no significant changes on the protein pattern of lentils germinated in the presence of elicitors were observed. Chitosan/glutamic acid increased by 1.6-fold the GABA content in lentil sprouts, whilst ascorbic and folic acids as well as chitosan/glutamic acid were highly effective to enhance the total content of phenolic compounds and the antioxidant activity of sprouted lentils. All elicited lentil sprouts showed ability to inhibit ACE activity (IC50: 9.5-11.9 µg peptides/mL). Therefore, elicitation can be considered a promising approach to improve the content of compounds with antioxidant and potential antihypertensive activities in lentil sprouts.

Synthesis of [(77)Se]-methylselenocysteine when preparing sauerkraut in the presence of [(77)Se]-selenite. Metabolic transformation of [ (77)Se]-methylselenocysteine in Wistar rats determined by LC-IDA-ICP-MS.

The use of enriched Se isotopes as tracers has provided important information on Se metabolism. However, selenium isotopes are expensive and difficult to obtain. A simple and cheap strategy based on the production of [(77)Se]-methylselenocysteine ([(77)Se]-MeSeCys) when preparing sauerkraut in the presence of [(77)Se]-selenite was developed. The resulting [(77)Se]-MeSeCys was used for evaluating the metabolic transformation of MeSeCys in Wistar rats, by feeding them with an AIN-93 M diet containing 20 % sauerkraut enriched in [(77)Se]-MeSeCys. Organs (liver, kidney, brain, testicles, and heart) were obtained after seven days of treatment and subjected to total selenium and selenium-speciation analysis by high-performance liquid chromatography coupled with isotope-dilution-analysis inductively-coupled-plasma mass spectrometry (HPLC-IDA-ICP-MS). Analysis of (77)Se-labeled organs revealed a prominent increase (more than 100 % Se-level enhancement) of selenium in the kidney and heart, whereas in the liver selenium concentration only increased by up to 20 % and it remained constant in the brain and testicles. (77)Se-enriched-sauerkraut supplementation does not alter the concentration of other essential elements in comparison to controls except for in the heart and kidney, in which selenium was positively correlated with Mg, Zn, Cu, and Mo. HPLC-ICP-MS analysis of hydrolyzed extracts after carbamidomethylation of the (77)Se-labeled organs revealed the presence of [(77)Se]-SeCys and an unknown Se-containing peak, the identity of which could not be verified by electrospray-ionization (ESI)-MS-MS. Low amounts of [(77)Se]-MeSeCys were found in (77)Se-labeled liver and kidney extracts, suggesting the incorporation of this selenium species in its intact form.

Biochemical and immunochemical evidences supporting the inclusion of quinoa (Chenopodium quinoa Willd.) as a gluten-free ingredient.

To date, the only acceptable therapeutic approach for celiac disease (CD) is a strict elimination from the diet of gluten-containing foods, but this diet does not always guarantee an adequate nutritional intake. Pseudocereals are receiving considerable attention as interesting alternatives for the formulation of gluten-free products, and quinoa grains arise as nutritive substitutes of conventional cereals. The aim of this study was the characterization of different quinoa samples corresponding to 11 quinoa varieties, using polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) and immunoblotting techniques to assess their suitability for celiac subjects. Some of these varieties were grown in Italy to assess if the reproduction in a new habitat can guarantee the retention of the "safe" protein pattern. None of the quinoa varieties studied presented protein bands with electrophoretic mobility comparable with those of wheat gliadins, the toxic protein for celiac subjects. All the quinoa samples showed a low binding affinity for both specific anti-gliadin antibodies and IgAs from celiac subjects, confirming that quinoa can be considered as a safe ingredient for celiac patients. However, reliable varieties should be previously selected since the immuno cross-reactivity with anti-gliadin antibodies can vary significantly.

Monitoring Mycotoxin Exposure in Food-Producing Animals (Cattle, Pig, Poultry, and Sheep).

Food-producing animals are exposed to mycotoxins through ingestion, inhalation, or dermal contact with contaminated materials. This exposure can lead to serious consequences for animal health, affects the cost and quality of livestock production, and can even impact human health through foods of animal origin. Therefore, controlling mycotoxin exposure in animals is of utmost importance. A systematic literature search was conducted in this study to retrieve the results of monitoring exposure to mycotoxins in food-producing animals over the last five years (2019-2023), considering both external exposure (analysis of feed) and internal exposure (analysis of biomarkers in biological matrices). The most commonly used analytical technique for both approaches is LC-MS/MS due to its capability for multidetection. Several mycotoxins, especially those that are regulated (ochratoxin A, zearalenone, deoxynivalenol, aflatoxins, fumonisins, T-2, and HT-2), along with some emerging mycotoxins (sterigmatocystin, nivalenol, beauvericin, enniantins among others), were studied in 13,818 feed samples worldwide and were typically detected at low levels, although they occasionally exceeded regulatory levels. The occurrence of multiple exposure is widespread. Regarding animal biomonitoring, the primary objective of the studies retrieved was to study mycotoxin metabolism after toxin administration. Some compounds have been suggested as biomarkers of exposure in the plasma, urine, and feces of animal species such as pigs and poultry. However, further research is required, including many other mycotoxins and animal species, such as cattle and sheep.

High pressure-assisted enzymatic hydrolysis potentiates the production of quinoa protein hydrolysates with antioxidant and ACE-inhibitory activities.

The impact of different pressure levels in the HHP-assisted hydrolysis by Alcalase of quinoa proteins on the catalytic efficiency, peptide release, phenolic compounds content, and biological activities was investigated. The protein profile (SDS-PAGE) showed a more extensive peptide breakdown for the HHP-assisted proteolysis at 300-400 MPa, which was confirmed by the higher extent of hydrolysis and peptide concentration. Quinoa protein hydrolysates (QPH) produced at 200 and 300 MPa exhibited higher total phenolic contents and antioxidant activities (methanol-acetone and aqueous extracts) when compared to the non-hydrolyzed (QPI) and non-pressurized hydrolyzed samples. Kaempferol dirhamnosyl-galactopyranoside was the prevalent phenolic compound in those samples, increasing total flavonoids by 1.8-fold over QPI. The QPH produced at 300 MPa inhibited ACE more effectively, exhibiting the greatest anti-hypertensive potential, along with the presence of several ACE-inhibitory peptides. The peptide sequences GSHWPFGGK, FSIAWPR, and PWLNFK presented the highest Peptide Ranker scores and were predicted to have ACE inhibitory, DPP-IV inhibitory, and antioxidant activities. Mild pressure levels were effective in producing QPH with enhanced functionality due to the effects of bioactive soluble phenolics and low molecular weight peptides.

High-Performance Liquid Chromatography-Fluorescence Detection Method for Ochratoxin A Quantification in Small Mice Sample Volumes: Versatile Application across Diverse Matrices Relevant for Neurodegeneration Research.

Ochratoxin A (OTA) is a mycotoxin commonly found in various food products, which poses potential health risks to humans and animals. Recently, more attention has been directed towards its potential neurodegenerative effects. However, there are currently no fully validated HPLC analytical methods established for its quantification in mice, the primary animal model in this field, that include pivotal tissues in this area of research, such as the intestine and brain. To address this gap, we developed and validated a highly sensitive, rapid, and simple method using HPLC-FLD for OTA determination in mice tissues (kidney, liver, brain, and intestine) as well as plasma samples. The method was rigorously validated for selectivity, linearity, accuracy, precision, recovery, dilution integrity, carry-over effect, stability, and robustness, meeting the validation criteria outlined by FDA and EMA guidelines. Furthermore, the described method enables the quantification of OTA in each individual sample using minimal tissue mass while maintaining excellent recovery values. The applicability of the method was demonstrated in a repeated low-dose OTA study in Balb/c mice, which, together with the inclusion of relevant and less common tissues in the validation process, underscore its suitability for neurodegeneration-related research.

Characterization of the sensitization profile to lupin in peanut-allergic children and assessment of cross-reactivity risk.

BACKGROUND: Case reports of allergy to lupin, due to primary sensitization or cross-reactions with other legumes, are increasing as a consequence of the augmented use of lupin flour in bakery, pasta formulations and other food items. The main allergens that have been associated with the sensitization to lupin are α- and ß-conglutins and, to a lesser extent, γ- and δ-conglutin, but no conclusive data are available so far. The aim of this study was to characterize the sensitization pattern to lupin in a group of 12 Italian children allergic to peanut and identify the specific lupin proteins involved in the cross-reactivity with peanut. METHODS: The immunochemical cross-reactivity among peanut and lupin was evaluated by both in vitro immunoblotting and in vivo fresh food skin prick test (FFSPT). RESULTS: The results showed that ß-conglutin was recognized by cutaneous IgEs from 7/12 peanut-allergic children in FFSPT and serum IgEs from 5/12 in immunoblotting, while 4/12 and 8/12 patients tested positive to γ-conglutin in FFSPT and immunoblotting, respectively. No significant immunoreactive responses were observed to α- and δ-conglutins under non-reducing conditions, but they were bound in FFSPT by the sera of 5/12 and 3/12 patients, respectively. CONCLUSION: In this group of allergic children, ß-conglutin has been identified as the major lupin allergen involved both in vitro and in vivo cross-reactivity with peanut proteins. The role of γ-conglutin in the cross-reactivity between lupin and peanut proteins was also relevant and clear, despite the observed unspecificity of the immunoblotting responses.

Impact of storage under ambient conditions on the vitamin content of dehydrated vegetables.

The consumption of dehydrated vegetables, which provides an important source of vitamins, is increasing worldwide. Dehydrated vegetables are located on non-refrigerated shelves in food shops and, therefore, it is of utmost importance to understand the modifications that take place in the content of these labile micronutrients at the ambient conditions currently found in food shops. The present study discusses the effect of storage for 3, 6, 9 and 12 months on the content of thiamin and vitamin C in different commercial and pilot plant dehydrated garlic, onions, potatoes and carrots in darkness at room temperature under vacuum conditions. The content of ß-carotene under these conditions was also studied in dehydrated carrots. Thiamin remained stable over the first 3 months of storage (∼90% retention), while long-term storage led to larger losses (retention of 85% in garlic and 45% in commercial carrots after 12 months of storage). The content of vitamin C drastically decreased during the storage period and even disappeared in some dried onions and carrots following 12 months of storage. Storage for 6 months at ambient conditions preserved 80-90% of the ß-carotene content in dehydrated vegetables, while long-term storage led to significant ß-carotene degradation (retentions between 43 and 81%). These results suggest that vitamins are gradually lost during storage at the practical conditions in food shops and will thus provide relevant information concerning dried vegetables, so manufacturers may calculate shelf life under established storage conditions.

Innovative Processing Technologies for Developing Functional Ingredients and Food Products with Health Benefits from Grains.

Grains are dry seeds belonging to diverse crops, including cereals, pseudocereals and pulses [...].

Co-Occurrence of Mycotoxins in Feed for Cattle, Pigs, Poultry, and Sheep in Navarra, a Region of Northern Spain.

Mycotoxins, toxic compounds produced by fungi on raw materials, such as cereals, represent a serious health hazard. Animals are exposed to them mainly through the ingestion of contaminated feed. This study presents data about the presence and co-occurrence of nine mycotoxins: aflatoxins B1, B2, G1, and G2, ochratoxins A and B, zearalenone (ZEA), deoxynivalenol (DON), and sterigmatocystin (STER), in 400 samples of compound feed for cattle, pigs, poultry, and sheep (100 samples each) collected in Spain (2019-2020). Aflatoxins, ochratoxins, and ZEA were quantified using a previously validated HPLC method using fluorescence detection; whereas DON and STER were quantified using ELISA. Moreover, the obtained results were compared with those obtained in this country and published in the last 5 years. The mycotoxin presence in Spanish feed, especially for ZEA and DON, has been demonstrated. The maximum individual levels found were: AFB1: 6.9 µg/kg in a sample of feed for poultry; OTA: 65.5 µg/kg in a sample of feed for pigs, DON: 887 µg/kg in a sample of feed for sheep, and ZEA: 816 µg/kg in a sample of feed for pigs. Nevertheless, regulated mycotoxins appear, in general, at levels below those regulated by the EU; in fact, the percentage of samples containing concentrations above these limits was very low (from 0% for DON to 2.5% for ZEA). The co-occurrence of mycotoxins has also been demonstrated: 63.5% of the analyzed samples presented detectable levels of two to five mycotoxins. Due to the fact that the distribution of mycotoxins in raw materials can change greatly from year to year with climate conditions or market globalization, regular mycotoxin monitorization in feed is needed to prevent the integration of contaminated materials in the food chain.

Biomonitoring of 19 Mycotoxins in Plasma from Food-Producing Animals (Cattle, Poultry, Pigs, and Sheep).

Mycotoxins are of great concern in relation to food safety. When animals are exposed to them, health problems, economic losses in farms and related industries, and the carryover of these compounds to animal-derived foods can occur. Therefore, control of animal exposure is of great importance. This control may be carried out by analyzing raw material and/or feed or through the analysis of biomarkers of exposure in biological matrixes. This second approach has been chosen in the present study. Firstly, a methodology capable of analyzing mycotoxins and some derivatives (AFB1, OTA, ZEA, DON, 3- and 15-ADON, DOM-1, T-2, HT-2, AFM1, STER, NEO, DAS, FUS-X, AFB2, AFG1, AFG2, OTB, and NIV) by LC-MS/MS in human plasma, has been revalidated to be applied in animal plasma. Secondly, this methodology was used in 80 plasma samples obtained from animals dedicated to food production: cattle, pigs, poultry, and sheep (20 samples of each), with and without being treated with a mixture of ß-glucuronidase-arylsulfatase to determine possible glucuronide and sulfate conjugates. Without enzymatic treatment, no mycotoxin was detected in any of the samples. Only one sample from poultry presented levels of DON and 3- and 15-ADON. With enzymatic treatment, only DON (1 sample) and STER were detected. The prevalence of STER was 100% of the samples, without significant differences among the four species; however, the prevalence and levels of this mycotoxin in the previously analyzed feed were low. This could be explained by the contamination of the farm environment. Animal biomonitoring can be a useful tool to assess animal exposure to mycotoxins. However, for these studies to be carried out and to be useful, knowledge must be increased on appropriate biomarkers for each mycotoxin in different animal species. In addition, adequate and validated analytical methods are needed, as well as knowledge of the relationships between the levels found in biological matrices and mycotoxin intake and toxicity.

In Vivo Genotoxicity and Toxicity Assessment of Sterigmatocystin Individually and in Mixture with Aflatoxin B1.

Mycotoxins are natural food and feed contaminants produced by several molds. The primary mode of exposure in humans and animals is through mixtures. Aflatoxin B1 (AFB1) and sterigmatocystin (STER) are structurally related mycotoxins that share the same biosynthetic route. Few in vivo genotoxicity assays have been performed with STER. In the present genotoxicity study, Wistar rats were dosed orally with STER (20 mg/kg b.w.), AFB1 (0.25 mg/kg b.w.) or a mixture of both in an integrated micronucleus (bone marrow) and comet study (liver and kidney). STER was dosed at the highest feasible dose in corn oil. No increase in the percentage of micronuclei in bone marrow was observed at any condition. Slight DNA damage was detected in the livers of animals treated with AFB1 or the mixture (DNA strand breaks and Fpg (Formamidopyrimidine DNA glycosylase)-sensitive sites, respectively). Plasma, liver, and kidney samples were analyzed with LC-MS/MS demonstrating exposure to both mycotoxins. General toxicity parameters (organs absolute weight, biochemistry, and histopathology) were not altered either individually or in the mixture. The overall absence of individual genotoxicity did not allow us to set any type of interaction in the mixture. However, a possible toxicokinetic interaction was observed.