RESUMEN
The objective of the present study was to explore the influence of dietary supplementation with a mixed additive (MA) containing a probiotic and anti-mycotoxin (Saccharomyces cerevisiae RC016 and Lactobacillus rhamnosus RC007) and its interaction on the performance and health (biochemistry and liver/intestine histopathology) of broilers fed diets contaminated with aflatoxin B1 (AFB1) at 506000±22.1ng/kg. The MA contained S. cerevisiae RC016 (1×107cells/g) and L. rhamnosus RC007 (1×108cells/g) in relation 1:1. A total of sixty-one-day-old Cobb broilers were randomly allocated into four treatment groups with three replicates of 5 birds each for a five-week-old feeding experiment. The experimental diet for each treatment (T) was formulated as follows: T1, a commercial diet (CD); T2, CD+AFB1; T3, CD+0.1% MA; T4, CD+AFB1+0.1% MA. The MA improved (p<0.01) production parameters (weight gain, conversion rate, and carcass yield) and reduced (p<0.01) the toxic effect of AFB1 on the relative weight of the livers. In addition, the macro and microscopic alterations of livers and the possible intestinal injury related to histological damage in the presence of mycotoxin were reduced. The use of probiotic MA based on S. cerevisiae RC016 and L. rhamnosus RC007 in animal feed provides greater protection against mycotoxin contamination and is safe for use as a supplement in animal feed, providing beneficial effects that improve animal health and productivity. This is of great importance at the economic level for the avian production system.
Asunto(s)
Aflatoxina B1 , Alimentación Animal , Pollos , Contaminación de Alimentos , Lacticaseibacillus rhamnosus , Probióticos , Saccharomyces cerevisiae , Animales , Aflatoxina B1/toxicidad , Pollos/microbiología , Suplementos Dietéticos , Hígado/efectos de los fármacos , Hígado/patologíaRESUMEN
Aflatoxin B1 (AFB1) and fumonisin B1 (FB1) are mycotoxins widely found as cereal contaminants, and their co-consumption is associated with liver cancer. Both are immunotoxic, but their interactions have been little studied. This work was aimed to evaluate in mouse spleen mononuclear cells (SMC) the effects of the exposure to AFB1 (5-50 µM), FB1 (25-250 µM), and AFB1-FB1 mixtures (MIX) on the in vitro differentiation of regulatory T cells (Treg and Tr1-like) and Th17 cells, as well as elucidate the contribution of aryl hydrocarbon receptor (Ahr) in such effects. AFB1 and mainly MIX induced cytotoxicity in activated CD4 cells via Ahr signaling. AFB1 (5 µM) increased the Treg cell differentiation, but its combination with FB1 (25 µM) also reduced Th17 cell expansion by Ahr-dependent mechanisms. Therefore, this mixture could enhance the Treg/Th17 cell ratio and favor immunosuppression and escape from tumor immunosurveillance to a greater extent than individual mycotoxins. Whereas, AFB1-FB1 mixtures at medium-high doses inhibited the Tr1-like cell expansion induced by the individual mycotoxins and affected Treg and Th17 cell differentiation in Ahr-independent and dependent manners, respectively, which could alter anti-inflammatory and Th17 immune responses. Moreover, individual FB1 altered regulatory T and Th17 cell development independently of Ahr. In conclusion, AFB1 and FB1 interact by modifying Ahr signaling, which is involved in the immunotoxicity as well as in the alteration of the differentiation of Treg, Tr1-like, and Th17 cells induced by AFB1-FB1 mixtures. Therefore, Ahr is implicated in the regulation of the anti- and pro-inflammatory responses caused by the combination of AFB1 and FB1.
Asunto(s)
Aflatoxina B1 , Diferenciación Celular , Fumonisinas , Receptores de Hidrocarburo de Aril , Linfocitos T Reguladores , Células Th17 , Receptores de Hidrocarburo de Aril/metabolismo , Aflatoxina B1/toxicidad , Animales , Células Th17/efectos de los fármacos , Linfocitos T Reguladores/efectos de los fármacos , Fumonisinas/toxicidad , Ratones , Diferenciación Celular/efectos de los fármacosRESUMEN
Mycotoxins, produced by fungi, can contaminate fish food and harm their health. Probiotics enhance immune balance and primarily function in the animal intestine. This study aimed to assess aflatoxin's impact on Piaractus mesopotamicus and explore probiotic-based additive (PBA) benefits in mitigating these effects, focusing on antioxidant activity, biochemical indices, and hepatic histopathology. Two experiments were conducted using P. mesopotamicus fry. The first experimental assay tested various levels of aflatoxin B1 (0.0, 25.0, 50.0, 100.0, 200.0, and 400.0 µg kg-1) over a 10-day period. The second experimental assay examined the efficacy of the probiotic (supplemented at 0.20%) in diets with different levels of aflatoxin B1 (0.0, 25.0, and 400.0 µg kg-1) for 15 days. At the end of each assay, the fish underwent a 24-hour fasting period, and the survival rate was recorded. Six liver specimens from each treatment group were randomly selected for metabolic indicator assays, including superoxide dismutase, catalase, alanine aminotransferase, aspartate aminotransferase, and albumin. Additionally, histopathological analysis was performed on six specimens. The initial study discovered that inclusion rates above 25.0 µg kg-1 resulted in decreased activity of AST (aspartate aminotransferase), ALT (alanine aminotransferase), ALB (albumin), CAT (catalase), and SOD (superoxide dismutase), accompanied by liver histopathological lesions. In the second study, the inclusion of PBA in diets contaminated with AFB1 improved the activity of AST and ALT up to 25.0 µg kg-1 of AFB1, with no histopathological lesions observed. The study demonstrated the hepatoprotective effects of PBA in diets contaminated with AFB1. The enzyme activity and hepatic histopathology were maintained, indicating a reduction in damage caused by high concentrations of AFB1 (400.0 µg kg-1 of AFB1). The adverse effects of AFB1 on biochemical and histopathological parameters were observed from 25.0 µg kg-1 onwards. Notably, PBA supplementation enhanced enzymatic activity at a concentration of 25 µg kg-1 of AFB1 and mitigated the effects at 400.0 µg kg-1 of AFB1. The use of PBAs in pacu diets is highly recommended as they effectively neutralize the toxic effects of AFB1 when added to diets containing 25.0 µg kg-1 AFB1. Dietary inclusion of aflatoxin B1 at a concentration of 25.0 µg kg-1 adversely affects the liver of Piaractus mesopotamicus (Pacu). However, the addition of a probiotic-based additive (PBA) to the diets containing this concentration of aflatoxin neutralized its toxic effects. Therefore, the study recommends the use of PBAs in Pacu diets to mitigate the adverse effects of aflatoxin contamination.
Asunto(s)
Aflatoxina B1 , Alimentación Animal , Enfermedades de los Peces , Hígado , Probióticos , Animales , Probióticos/farmacología , Probióticos/administración & dosificación , Hígado/efectos de los fármacos , Hígado/patología , Alimentación Animal/análisis , Enfermedades de los Peces/inducido químicamente , Enfermedades de los Peces/prevención & control , Enfermedades de los Peces/patología , Aflatoxina B1/toxicidad , Dieta/veterinaria , Suplementos Dietéticos/análisis , Aflatoxinas/toxicidadRESUMEN
This experiment was conducted to determine the effect of an adsorbent material based on powdered alfalfa leaves added in the aflatoxin B1 (AFB1)-contaminated diet of turkey poults on production parameters, blood cell count, serum biochemistry, liver enzymes, and liver histology. For this purpose, three hundred and fifty female Nicholas-700 poults were randomly assigned into five treatments: (1) Control, AFB1-free diet; (2) AF, diet contaminated with 250 ng AFB1/g; (3) Alfalfa, AFB1-free diet + 0.5% (w/w) adsorbent; (4) AF+alfalfa, diet contaminated with 250 ng AFB1/g + 0.5% (w/w) adsorbent, and (5) AF+ yeast cell wall (YCW), diet contaminated with 250 ng AFB1/g + 0.5% (w/w) of yeast cell wall (a commercial mycotoxin binder used as reference material). The in vivo efficacy of powdered alfalfa leaves was assessed during a 28-day period. In general, the addition of powdered alfalfa leaves in the AFB1-free diet gave the best performance results (body weight, body weight gain, and feed intake) and improved the values of total protein, glucose, calcium, creatinine, and blood urea nitrogen. Moreover, the addition of powdered alfalfa leaves in the AFB1-contaminated diet enhanced body weight and body weight gain and significantly reduced the feed intake, compared to the AF and AF+YCW groups. Additionally, significant alterations in serum parameters were observed in poults intoxicated with the AFB1, compared to the Control group. Furthermore, typical histopathological lesions were observed in the liver of the AF group, which were significantly ameliorated with the addition of powdered alfalfa leaves. Conclusively, these results pointed out that low inclusion of powdered alfalfa leaves in the contaminated feed counteracted the adverse effects of AFB1 in turkey poults.
Asunto(s)
Aflatoxina B1 , Alimentación Animal , Medicago sativa , Hojas de la Planta , Pavos , Animales , Aflatoxina B1/toxicidad , Medicago sativa/química , Hojas de la Planta/química , Alimentación Animal/análisis , Femenino , Hígado/efectos de los fármacos , Hígado/patología , Dieta/veterinaria , Polvos , Peso Corporal/efectos de los fármacosRESUMEN
The present study aimed to evaluate whether a moderate dose of aflatoxin B1 in pigs' diet interferes with pigs' growth and health in the nursery phase and whether an anti-mycotoxin mixture minimizes the adverse effects of the toxin. One blend with Saccharomyces cerevisiae lysate, zeolite, silicon dioxide, propylene glycol, Carduus marianus extract, soy lecithin, and carbonate was used as an anti-mycotoxin. Four treatments, with six repetitions per treatment and three pigs/pen: Afla0-AntiMyc0 - negative control (without aflatoxin); Afla500-AntiMyc0 - positive control (500 ppb of aflatoxin); Afla0-AntiMyc1000 - 1000 mg/kg of anti-mycotoxin blend; Afla500-AntiMyc1000 - 500 ppb aflatoxin +1000 mg/kg of anti-mycotoxin blend. It was observed that pigs in the positive control (Afla500-AntiMyc0) had lower body weight and weight gain when compared to the other treatments during the experimental period. Also, pigs from Afla500-AntiMyc0 had lower feed intake between days 1-20 and 1 to 30 than Afla0-AntiMyc0. The pigs from Afla500-AntiMyc0 had higher levels of liver enzymes aspartate aminotransferase and alanine aminotransferase compared to other treatments. The pigs from Afla500-AntiMyc0 had higher villus height than the other treatments, while the folded size was smaller in this treatment. Crypts were deeper in the intestines of pigs in both treatments that consumed aflatoxin. In general, it is concluded that the intake of aflatoxin B1 by nursery pigs has negative impacts on the health and, consequently, the animals' growth performance; however, the addition of the contaminated feed with an anti-mycotoxin blend was able to protect the pigs, minimizing the adverse effects caused by the mycotoxin.
Asunto(s)
Aflatoxina B1 , Micotoxinas , Porcinos , Animales , Aflatoxina B1/toxicidad , Aspergillus flavus , Dieta/veterinaria , Aumento de Peso , Alimentación Animal/análisisRESUMEN
The objectives of this study were to evaluate the exposure to a diet naturally contaminated with mycotoxins on lactation performance, animal health, and the ability to sequester agents (SA) to reduce the human exposure to AFM1. Sixty healthy lactating Holstein cows were randomly assigned to two groups: naturally contaminated diet without and with the addition of a SA (20 g/cow/d AntitoxCooPil® -60% zeolite-40% cell wall-). Each cow was monitored throughout lactation. The concentration of aflatoxin B1 (AFB1) in feed and M1 (AFM1) in milk, health status, and productive and reproductive parameters were measured. AFB1 concentration in feed was very low (2.31 µg/kgDM). The addition of SA reduced the milk AFM1 concentrations (0.016 vs. 0.008 µg/kg) and transfer rates (2.19 vs. 0.77%). No differences were observed in health status, production and reproduction performance. The inclusion of SA in the diet of dairy cows reduce the risk in the most susceptible population.
Asunto(s)
Aflatoxina M1 , Contaminación de Alimentos , Lactancia , Leche , Secuestrantes , Animales , Bovinos , Femenino , Aflatoxina B1/toxicidad , Aflatoxina B1/análisis , Aflatoxina M1/análisis , Aflatoxina M1/antagonistas & inhibidores , Alimentación Animal/análisis , Alimentación Animal/toxicidad , Dieta/veterinaria , Contaminación de Alimentos/análisis , Contaminación de Alimentos/prevención & control , Leche/química , Secuestrantes/administración & dosificación , Distribución AleatoriaRESUMEN
Abstract Lactobacilli are probiotics with Aflatoxin (AF) detoxification ability, found in fermented products, GIT of animals and environment. Purpose of this study was to investigate the ability of broiler isolates of Lactobacillus against Aflatoxin B1 (AFB1). For this purpose, 5 isolates of Lactobacillus from broiler gut were incubated with 100 ppb AFB1 in aqueous environment and effect of different parameters (cell fractions, time, temperature, pH) on detoxification was determined by HPLC. The ameliorative effect of Lactobacillus salivarius (LS) against AFB1 was studied in broiler. The results revealed that LS (CR. 4) showed the best results (in vitro) as compared to other isolates (L. salivarius (CR. 3, CR, 4), L. agilis (CE. 2.1, CE. 3.1) and L. crispatus (CE. 28). Cell debris of CR. 4 showed significantly higher detoxification (P<0.05). Maximum amount of AFB1 was detoxified at 30°C (97%), pH 4.0 (99%) and 6 h (99.97%). In vivo study showed that AFB1 decreased weight gain (1,269 ± 0.04 gm/ bird), feed consumed (2,161 ± 0.08 gm/ bird), serum total protein (2.42 ± 0.34 gm/ dl), serum albumin (0.5 ± 0.2 2 gm/dl) and antibody titer (4.2 ± 0.83). Liver function enzymes were found (alanine transaminase (ALT): 32 ± 10.7 U/L) and aspartate transaminase (AST): 314.8 ± 27 U/L) elevated in AFB1 fed broilers. Treatment with 1% LS not only decreased the toxic effects of AFB1 (group D) but also improved the overall health of broilers due to its probiotic effects (p<0.05) as compared to control negative (group A). The detoxification ability of LS was better than commercial binder (CB) (0.2% Protmyc). It was concluded that detoxification of AFB1 by Lactobacillus was strain, temperature, pH and time dependent. LS has detoxification ability against AFB1 in vivo.
Resumo Os lactobacilos são probióticos com capacidade de desintoxicação da Aflatoxina (AF), encontrados em produtos fermentados, TGI de animais e meio ambiente. O objetivo deste estudo foi investigar a capacidade de isolados de frango de corte de Lactobacillus contra a Aflatoxina B1 (AFB1). Para tanto, 5 isolados de Lactobacillus de intestino de frango foram incubados com 100 ppb AFB1 em meio aquoso, e o efeito de diferentes parâmetros (frações celulares, tempo, temperatura, pH) na desintoxicação foi determinado por CLAE. O efeito melhorador de Lactobacillus salivarius (LS) contra AFB1 foi estudado em frangos de corte. Os resultados revelaram que LS (CR. 4) apresentou os melhores resultados (in vitro) em comparação com outros isolados [L. salivarius (CR. 3, CR. 4), L. agilis (CE. 2.1, CE. 3.1) e L. crispatus (CE. 28)]. Detritos celulares de CR. 4 mostraram desintoxicação significativamente maior (P < 0.05). A quantidade máxima de AFB1 foi desintoxicada a 30 °C (97%), pH 4.0 (99%) e 6 h (99,97%). O estudo in vivo mostrou que AFB1 diminuiu o ganho de peso (1,269 ± 0.04 g / ave), alimento consumido (2,161 ± 0.08 g / ave), proteína total sérica (2.42 ± 0.34 g / dl), albumina sérica (0.5 ± 0.22 gm / dl) e título de anticorpo (4.2 ± 0.83). As enzimas da função hepática foram encontradas (alanina transaminase (ALT): 32 ± 10.7 U / L) e aspartato transaminase (AST): 314.8 ± 27 U / L) elevadas em AFB1 alimentados com frangos. O tratamento com 1% LS não só diminuiu os efeitos tóxicos de AFB1 (grupo D), mas também melhorou a saúde geral dos frangos devido aos seus efeitos probióticos (p < 0.05) em comparação com o controle negativo (grupo A). A capacidade de desintoxicação do LS foi melhor do que o aglutinante comercial (CB) (0.2% Protmyc). Concluiu-se que a desintoxicação de AFB1 por Lactobacillus foi dependente da cepa, temperatura, pH e tempo. LS tem capacidade de desintoxicação contra AFB1 in vivo.
Asunto(s)
Animales , Aflatoxina B1/análisis , Aflatoxina B1/toxicidad , Probióticos , Pollos , Lactobacillus , Alimentación Animal/análisisRESUMEN
Introduction: Aflatoxins B1 are among the most common poisonous mycotoxins produced by certain fungi that harm animals and crops. Mycotoxins can cause a variety of adverse health effects and pose a serious health threat to humans. The Maximum Residue Limits of aflatoxin B1 in processed cereals and ingredients are 2 parts per billion (ppb) and 5 ppb, respectively. Objectives: To evaluate the status of aflatoxin B1 contamination in rice, corn and staple food produced in Ha Giang province compared with the maximum permitted levels. Methods: A total of 210 rice and maize samples were analyzed to quantify the level of aflatoxin B1. Analysis of mycotoxins was conducted by High Performance Liquid Chromatography using a fluorescence detector. Results: It was found that rice, rice products, maize, and maize products had a mean aflatoxin B1 content of 1.79 ppb, 2.55 ppb, 2.19 ppb, and 6.35 ppb, respectively. The results also showed that 71.9 percent of samples were contaminated with mycotoxins, and 14.28 percent of samples exceeded the maximum allowable limit. Conclusion: The concentration of aflatoxin B1 in 14.28 percent of the samples are over permissible limits by nationwide regulations (AU)
Introducción: La aflatoxina B1 se encuentra entre las micotoxinas más comunes y venenosas producidas por ciertos hongos que dañan a los animales y los cultivos. Las micotoxinas pueden causar una variedad de efectos adversos para la salud y representar una grave amenaza para la salud de los seres humanos. Los límites máximos de residuos de aflatoxina B1en cereales e ingredientes procesados son de 2 ppb y 5 ppb, respectivamente. Objetivos: Evaluar el estado de contaminación por aflatoxina B1 en arroz, maíz y alimentos básicos producidos en la provincia de Ha Giang, en comparación con los niveles máximos permitidos. Métodos: Se analizaron un total de 210 muestras de arroz y maíz para cuantificar el nivel de aflatoxina B1. El análisis de micotoxinas se realizó mediante cromatografía líquida de alta resolución, utilizando un detector de fluorescencia. Resultados: Se encontró que el arroz, los productos de arroz, el maíz y los productos de maíz tenían un contenido medio de aflatoxin B1, de 1,79 ppb, 2,55 ppb, 2,19 ppb y 6,35 ppb, respectivamente. Los resultados también mostraron que el 71,9 por ciento de las muestras estaban contaminadas con micotoxinas y el 14,28 por ciento de las muestras excedieron el límite máximo permitido. Conclusión: La concentración de aflatoxina B1 en el 14,28 por ciento de las muestras está por encima de los límites permisibles por la norma nacional(AU)
Asunto(s)
Oryza , Contaminación de Alimentos , Cromatografía Líquida de Alta Presión/métodos , Aflatoxina B1/toxicidad , Zea mays , Micotoxinas/análisis , Producción de Cultivos/métodosRESUMEN
Aflatoxin B1 (AFB1) exhibits the most potent mutagenic and carcinogenic activity among aflatoxins. For this reason, AFB1 is recognized as a human group 1 carcinogen by the International Agency of Research on Cancer. Consequently, it is essential to determine its properties and behavior in different chemical systems. The chemical properties of AFB1 can be explored using computational chemistry, which has been employed complementarily to experimental investigations. The present review includes in silico studies (semiempirical, Hartree-Fock, DFT, molecular docking, and molecular dynamics) conducted from the first computational study in 1974 to the present (2022). This work was performed, considering the following groups: (a) molecular properties of AFB1 (structural, energy, solvent effects, ground and the excited state, atomic charges, among others); (b) theoretical investigations of AFB1 (degradation, quantification, reactivity, among others); (c) molecular interactions with inorganic compounds (Ag+, Zn2+, and Mg2+); (d) molecular interactions with environmentally compounds (clays); and (e) molecular interactions with biological compounds (DNA, enzymes, cyclodextrins, glucans, among others). Accordingly, in this work, we provide to the stakeholder the knowledge of toxicity of types of AFB1-derivatives, the structure-activity relationships manifested by the bonds between AFB1 and DNA or proteins, and the types of strategies that have been employed to quantify, detect, and eliminate the AFB1 molecule.
Asunto(s)
Aflatoxina B1 , Aflatoxinas , Humanos , Aflatoxina B1/toxicidad , Simulación del Acoplamiento Molecular , Aflatoxinas/metabolismo , Relación Estructura-Actividad , Carcinógenos , ADN/metabolismoRESUMEN
Several studies demonstrated the toxicity of aspartame (ASP) and aflatoxin B1 (AFB1 ) in preclinical models. Although the majority of these reports assessed the toxic effects of each substance separately, their concomitant exposure and hazardous consequences are scarce. Importantly, the deleterious effects at the central nervous system caused by ASP and AFB1 co-exposure are rarely addressed. We evaluated if concomitant exposure to AFB1 and ASP would cause behavioral impairment and alteration in oxidative status of the brain in male rats. Animals received once a day for 14 days AFB1 (250 µg/kg, intragastric gavage [i.g.]), ASP (75 mg/kg, i.g.), or both substances (association). On day 14, they were subjected to behavioral evaluation, and biochemical and molecular parameters of oxidative status were measured in the cerebral cortex and hippocampus. In the open field test, AFB1 and combination treatments modified the motor, exploratory, and grooming behavior. In the splash test, all treatments caused a reduction in grooming time compared to the control group. An increase in thiobarbituric acid-reactive substances content induced by AFB1 and combination treatments was observed. The antioxidant defenses (vitamin C, nonprotein sulfhydryl, and ferric reducing antioxidant power) were impaired in all groups compared to control. Regarding molecular evaluation, mitochondrial superoxide dismutase-2 immunoreactivity decreased after AFB1 or ASP exposition in the hippocampus. Thus, co-exposure to ASP and AFB1 was potentially more toxic because it aggravated behavioral impairments and oxidative status disbalance in comparison to the groups that received only ASP or AFB1 . Therefore, our data suggest that those substances caused a disruption in brain homeostasis.
Asunto(s)
Aflatoxina B1 , Antioxidantes , Ratas , Masculino , Animales , Antioxidantes/farmacología , Aflatoxina B1/toxicidad , Aspartame/toxicidad , Ácido Ascórbico/farmacología , Hipocampo/metabolismo , Estrés OxidativoRESUMEN
The potential interactions among food additives/contaminants and the consequences to biological systems is a topic that is rarely addressed in scientific literature. Thus, the current study investigated if the combined administration of ASP and AFB1 would impair hepatic and renal oxidative status. Male Wistar rats received during 14 days once a day ASP (75 mg/Kg) and/or AFB1 (250 µg/Kg) through intragastric route. At the end of experimental protocol, samples of liver and kidneys were collected for assessing biochemical markers of oxidative status. In the hepatic tissue, the treatment with a single substance (ASP or AFB1) caused an increase in TBARS levels, and a reduction in non-enzymatic antioxidant defenses (Vit C and NPSH levels and FRAP test). In the kidneys, TBARS levels were increased only in the group that received ASP + AFB1. The association reduced NPSH content, while the treatment with AFB1 reduced the FRAP levels. GST and CAT activities were increased in all treatments. Overall, ASP and AFB1 association presented higher toxic effects to the tissues. To the best of our knowledge, this is the first study demonstrating that the associated use of both ASP and AFB1 induces more extensive injuries in comparison to the effects caused by each one alone. Therefore, these data demonstrated that concomitant exposure to ASP and AFB1 potentiated their oxidative damage in hepatic tissue, suggesting that this organ is particularly sensitive to the toxic action induced by these substances.
Asunto(s)
Aflatoxina B1 , Antioxidantes , Ratas , Masculino , Animales , Aflatoxina B1/toxicidad , Antioxidantes/farmacología , Aspartame/toxicidad , Aspartame/metabolismo , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , Ratas Wistar , Estrés Oxidativo , Hígado , Biomarcadores/metabolismo , Aditivos Alimentarios/metabolismo , Aditivos Alimentarios/farmacologíaRESUMEN
In vitro tests are performed to evaluate the efficacy of antimycotoxins additives (AMAs); nevertheless, such assays show a low correlation with in vivo trials, which are also required to determine AMAs' efficacy. In search of an alternative method, the current study investigated the use of an ex vivo technique. Six AMAs (AMA1 to AMA6) had their ability to reduce intestinal absorption of aflatoxin B1 (AFB1) evaluated. Jejunal explants were obtained from broilers and subjected to two treatments per AMA in Ussing chambers: T1 (control) - 2.8 mg/L AFB1, and T2 - 2.8 mg/L AFB1 + 0.5% AMA. AMAs were also tested in vitro to assess adsorption of AFB1 in artificial intestinal fluid. In the ex vivo studies, AMA1 to AMA6 decreased intestinal absorption of AFB1 by 67.11%, 73.82%, 80.70%, 85.86%, 86.28% and 82.32%, respectively. As for the in vitro results, AMA1 to AMA6 presented an adsorption of 99.72%, 99.37%, 99.67%, 99.53%, 99.04% and 99.15%, respectively. The evaluated ex vivo model proved useful in the assessment of AMAs. No correlation was reported between ex vivo and in vitro findings. Further studies are needed to elucidate the correlation between ex vivo and in vivo results seeking to reduce animal testing.
Testes in vitro são realizados para avaliar a eficácia de aditivos antimicotoxinas (AAMs); entretanto, tais experimentos apresentam uma baixa correlação com ensaios in vivo, que também são exigidos para determinar a eficácia de AAMs. Em busca de um método alternativo, o presente estudo investigou o uso de uma técnica ex vivo. A capacidade de seis AAMs (AAM1 a AAM6) de reduzir a absorção intestinal de aflatoxina B1 (AFB1) foi avaliada. Explantes jejunais foram coletados de frangos de corte e submetidos a dois tratamentos por AAM em câmaras de Ussing: T1 (controle) - 2,8 mg/L AFB1, e T2 - 2.8 mg/L AFB1 + 0,5% AAM. Os AAMs também foram testados in vitro para verificar a adsorção de AFB1 em fluido intestinal artificial. Nos ensaios ex vivo, AAM1 ao AAM6 diminuíram a absorção intestinal de AFB1 em 67,11%, 73,82%, 80,70%, 85,86%, 86,28% e 82,32%, respectivamente. Quanto aos achados in vitro, AAM1 ao AAM6 apresentaram adsorção de 99,72%, 99,37%, 99,67%, 99,53%, 99,04% e 99,15%, respectivamente. O modelo ex vivo avaliado mostrou-se eficiente na avaliação de AAMs. Não houve correlação entre os resultados ex vivo e in vitro. Estudos adicionais são necessários para definir a correlação entre achados ex vivo e in vivo na tentativa de reduzir os testes em animais.
Asunto(s)
Animales , Antitoxinas/análisis , Pollos , Aflatoxina B1/toxicidad , Yeyuno , Técnicas In VitroRESUMEN
Lactobacilli are probiotics with Aflatoxin (AF) detoxification ability, found in fermented products, GIT of animals and environment. Purpose of this study was to investigate the ability of broiler isolates of Lactobacillus against Aflatoxin B1 (AFB1). For this purpose, 5 isolates of Lactobacillus from broiler gut were incubated with 100 ppb AFB1 in aqueous environment and effect of different parameters (cell fractions, time, temperature, pH) on detoxification was determined by HPLC. The ameliorative effect of Lactobacillus salivarius (LS) against AFB1 was studied in broiler. The results revealed that LS (CR. 4) showed the best results (in vitro) as compared to other isolates (L. salivarius (CR. 3, CR, 4), L. agilis (CE. 2.1, CE. 3.1) and L. crispatus (CE. 28). Cell debris of CR. 4 showed significantly higher detoxification (P<0.05). Maximum amount of AFB1 was detoxified at 30°C (97%), pH 4.0 (99%) and 6 h (99.97%). In vivo study showed that AFB1 decreased weight gain (1,269 ± 0.04 gm/ bird), feed consumed (2,161 ± 0.08 gm/ bird), serum total protein (2.42 ± 0.34 gm/ dl), serum albumin (0.5 ± 0.2 2 gm/dl) and antibody titer (4.2 ± 0.83). Liver function enzymes were found (alanine transaminase (ALT): 32 ± 10.7 U/L) and aspartate transaminase (AST): 314.8 ± 27 U/L) elevated in AFB1 fed broilers. Treatment with 1% LS not only decreased the toxic effects of AFB1 (group D) but also improved the overall health of broilers due to its probiotic effects (p<0.05) as compared to control negative (group A). The detoxification ability of LS was better than commercial binder (CB) (0.2% Protmyc). It was concluded that detoxification of AFB1 by Lactobacillus was strain, temperature, pH and time dependent. LS has detoxification ability against AFB1 in vivo.
Asunto(s)
Aflatoxina B1 , Probióticos , Aflatoxina B1/análisis , Aflatoxina B1/toxicidad , Alimentación Animal/análisis , Animales , Pollos , LactobacillusRESUMEN
Aflatoxins are mycotoxins produced as secondary fungal metabolites. Among them, aflatoxin B1 (AFB1) stands out due to its genotoxic and mutagenic potential, being a potent initiator of carcinogenesis. In this review, the outcomes from the published literature in the past 10 years on the effects of AFB1 pathophysiological mechanisms on embryological and fetal development are discussed. In several animal species, including humans, AFB1 has a teratogenic effect, resulting in bone malformations, visceral anomalies, lesions in several organs, and behavioral and reproductive changes, in addition to low birth weight. The mutagenic capacity of AFB1 in prenatal life is greater than in adults, indicating that when exposure occurs in the womb, the risk of the development of neoplasms is higher. Studies conducted in humans indicate that the exposure to this mycotoxin during pregnancy is associated with low birth weight, decreased head circumference, and DNA hypermethylation. However, as the actual impacts on humans are still unclear, the importance of this issue cannot be overemphasized and studies on the matter are essential.
Asunto(s)
Aflatoxina B1/toxicidad , Mutágenos/toxicidad , Efectos Tardíos de la Exposición Prenatal/inducido químicamente , Animales , Metilación de ADN/efectos de los fármacos , Femenino , Humanos , Neoplasias/inducido químicamente , Neoplasias/genética , Neoplasias/patología , Embarazo , Efectos Tardíos de la Exposición Prenatal/genética , Efectos Tardíos de la Exposición Prenatal/patologíaRESUMEN
Aflatoxin B1 (AFB1) is a mycotoxin highly toxic and carcinogenic to humans due to its potential to induce oxidative stress. The Beta-caryophyllene (BCP) have been highlighted for its broad spectrum of pharmacological effects. The present study aimed to investigate the beneficial effects of BCP against the susceptibility of hepatic and renal tissues to AFB1 toxicity, in biochemical parameters to assess organ function, tissue oxidation, and the immunocontent of oxidative and inflammatory proteins. Male Wistar rats was exposed to AFB1 (250 µg/kg, i.g.) and/or BCP (100 mg/kg, i.p.) for 14 successive days. It was found that exposure to AFB1 did not change the measured renal toxicity parameters. Also, AFB1 increased liver injury biomarkers (gamma glutamyl transferase and alkaline phosphatase) and reduced levels of non-enzymatic antioxidant defenses (ascorbic acid and non-protein thiol), however did not cause changes in the lipid peroxidation levels. Moreover, AFB1 interfered in oxidative pathway regulated by Kelch-like ECH-associated protein (Keap1)/nuclear factor (erythroid-derived 2)-like 2 (Nrf2), overacting Glutathione-S-Transferase (GST) activity. Lastly, a main effect of AFB1 on the total interleukin 1 beta (IL-1ß) was observed. Remarkably, the associated treatment of AFB1 + BCP improved altered liver parameters. In addition, BCP and AFB1 + BCP groups showed an increase in the levels of inhibitor of nuclear factor kappa-B kinase subunit beta (IKKß). Thus, these results indicated that BCP has potential protective effect against AFB1 induced hepatotoxicity.
Asunto(s)
Aflatoxina B1/toxicidad , Citoprotección/efectos de los fármacos , Hígado/efectos de los fármacos , Sesquiterpenos/farmacología , Animales , Antioxidantes/metabolismo , Glutatión/metabolismo , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Hígado/citología , Hígado/metabolismo , Masculino , Factor 2 Relacionado con NF-E2/metabolismo , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas WistarRESUMEN
In this study, the changes in oncogenic and tumor suppressor signaling pathways in liver and their association with serum and urinary biomarkers of aflatoxin exposure were evaluated in Wistar rats fed diets containing aflatoxin B1 (AFB1) for 90 days. Rats were divided into four groups (n = 15 per group) and assigned to dietary treatments containing 0 (control), 50 (AFB50), 100 (AFB100) and 200 µg AFB1 kg-1 diet (AFB200). Multiple preneoplastic foci of hepatocytes marked with glutathione-S-transferase-placental form (GST-P) were identified in AFB100 and AFB200 groups. Hepatocellular damage induced by AFB1 resulted in overexpression of cyclin D1 and ß-catenin. The liver expression of retinoblastoma (Rb) and p27Kip1 decreased in AFB100 and AFB200 groups, confirming the favorable conditions for neoplastic progression to hepatocellular carcinoma. All samples from rats fed AFB1-contaminated diets had quantifiable AFB1-lysine in serum or urinary AFM1 and AFB1-N7-guanine, with mean levels of 20.42-50.34 ng mL-1, 5.31-37.68 and 39.15-126.37 ng mg-1 creatinine, respectively. Positive correlations were found between AFB1-lysine, AFM1 or AFB1-N7-guanine and GST-P+, ß-catenin+ and cyclin D1+ hepatocytes, while Rb + cells negatively correlated with those AFB1 exposure biomarkers. The pathways evaluated are critical molecular mechanisms of AFB1-induced hepatocarcinogenesis in rats.
Asunto(s)
Aflatoxina B1/toxicidad , Ciclina D1/metabolismo , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Proteína de Retinoblastoma/metabolismo , beta Catenina/metabolismo , Aflatoxina B1/análogos & derivados , Aflatoxina B1/sangre , Aflatoxina B1/metabolismo , Aflatoxina B1/orina , Aflatoxina M1/orina , Alanina Transaminasa/metabolismo , Animales , Aspartato Aminotransferasas/metabolismo , Biomarcadores/sangre , Biomarcadores/orina , Expresión Génica/efectos de los fármacos , Guanina/análogos & derivados , Guanina/orina , Hepatocitos/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/patología , Lisina/sangre , Masculino , Lesiones Precancerosas/inducido químicamente , Lesiones Precancerosas/patología , Ratas WistarRESUMEN
Studies have shown that the intracellular content of probiotic (postbiotics) has antioxidant properties, which can improve the antioxidant status in vivo. However, its absorption and mechanisms underlying the protective effects are still unknown. The antioxidant capacity of Lacticaseibacillus casei CRL431 (IC-431) postbiotics was determined after an in vitro simulated digestive process. Permeability of antioxidant constituents of IC-431 was determined by an ex vivo everted duodenum assay. Aflatoxin B1-induced oxidative stress rat models were established and treated with IC-431; biomarkers of hepatic mitochondrial function and H2O2 levels, oxidative stress, and oxidative stress index (OSi) were examined. The antioxidant capacity of IC-431 (477 ± 45.25 µmol Trolox Equivalent/L) was reduced by exposure to the simulated digestive process. No difference (p > 0.05) was found among digested and the permeate fraction of IC-431. A protective effect was observed by significantly lower OSi and higher liver glutathione peroxidase and catalase activities. Lower H2O2 production, a higher degree of mitochondrial uncoupling, and lower mitochondrial respiration coefficient were also observed (p < 0.05). These results suggest that IC-431 antioxidant components permeate intestinal barriers to enter the bloodstream and regulate antioxidant status during AFB1-induced oxidative stress by reducing hepatic mitochondrial dysfunction, thus enhancing antioxidant enzyme response.
Asunto(s)
Aflatoxina B1 , Lacticaseibacillus casei , Mitocondrias , Estrés Oxidativo , Probióticos , Aflatoxina B1/toxicidad , Animales , Antioxidantes , Peróxido de Hidrógeno , Mitocondrias/fisiología , RatasRESUMEN
OBJECTIVE: In Guatemala, cirrhosis is among the 10 leading causes of death, and mortality rates have increased lately. The reasons for this heavy burden of disease are not clear as the prevalence of prominent risk factors, such as hepatitis B virus, hepatitis C virus and heavy alcohol consumption, appears to be low. Aflatoxin B1 (AFB1) exposure, however, appears to be high, and thus could be associated with the high burden of cirrhosis. Whether AFB1 increases the risk of cirrhosis in the absence of viral infection, however, is not clear. DESIGN: Cirrhosis cases (n=100) from two major referral hospitals in Guatemala City were compared with controls (n=200) from a cross-sectional study. Logistic regression was used to estimate the ORs and 95% CIs of cirrhosis and quintiles of AFB1 in crude and adjusted models. A sex-stratified analysis was also conducted. RESULTS: The median AFB1 level was significantly higher among the cases (11.4 pg/mg) than controls (5.11 pg/mg). In logistic regression analyses, higher levels of AFB1 was associated with cirrhosis (quintile 5 vs quintile 1, OR: 11.55; 95% CI 4.05 to 32.89). No attenuation was observed with adjustment by sex, ethnicity, hepatitis B virus status, and heavy alcohol consumption. A significantly increasing trend in association was observed in both models (p trend <0.01). Additionally, the cirrhosis-AFB1 association was more prominent among men. CONCLUSIONS: The current study found a significant positive association between AFB1 exposure and cirrhosis. Mitigation of AFB1 exposure and a better understanding of additional risk factors may be important to reduce the burden of cirrhosis in Guatemala.
Asunto(s)
Aflatoxina B1/sangre , Consumo Excesivo de Bebidas Alcohólicas/complicaciones , Cirrosis Hepática/etiología , Micotoxinas/sangre , Aflatoxina B1/efectos adversos , Aflatoxina B1/toxicidad , Consumo Excesivo de Bebidas Alcohólicas/epidemiología , Estudios de Casos y Controles , Costo de Enfermedad , Estudios Transversales , Exposición a Riesgos Ambientales , Femenino , Guatemala/epidemiología , Hepacivirus/aislamiento & purificación , Hepatitis B/complicaciones , Hepatitis B/epidemiología , Hepatitis B/virología , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis C/complicaciones , Hepatitis C/epidemiología , Hepatitis C/virología , Humanos , Cirrosis Hepática/epidemiología , Cirrosis Hepática/metabolismo , Cirrosis Hepática/mortalidad , Modelos Logísticos , Masculino , Persona de Mediana Edad , Micotoxinas/efectos adversos , Micotoxinas/toxicidad , Prevalencia , Factores de RiesgoRESUMEN
Aflatoxin B1 aldehyde reductase (AFAR) enzyme activity has been associated to a higher resistance to the aflatoxin B1 (AFB1) toxicity in ethoxyquin-fed rats. However, no studies about AFAR activity and its relationship with tolerance to AFB1 have been conducted in poultry. To determine the role of AFAR in poultry tolerance, the hepatic in vitro enzymatic activity of AFAR was investigated in liver cytosol from four commercial poultry species (chicken, quail, turkey and duck). Specifically, the kinetic parameters Vmax, Km and intrinsic clearance (CLint) were determined for AFB1 dialdehyde reductase (AFB1-monoalcohol production) and AFB1 monoalcohol reductase (AFB1-dialcohol production). In all cases, AFB1 monoalcohol reductase activity saturated at the highest aflatoxin B1 dialdehyde concentration tested (66.4 µM), whereas AFB1 dialdehyde reductase did not. Both activities were highly and significantly correlated and therefore are most likely catalyzed by the same AFAR enzyme. However, it appears that production of the AFB1 monoalcohol is favored over the AFB1 dialcohol. The production of alcohols from aflatoxin dialdehyde showed the highest enzymatic efficiency (highest CLint value) in chickens, a species resistant to AFB1; however, it was also high in the turkey, a species with intermediate sensitivity; further, CLint values were lowest in another tolerant species (quail) and in the most sensitive poultry species (the duck). These results suggest that AFAR activity is related to resistance to the acute toxic effects of AFB1 only in chickens and ducks. Genetic selection of ducks for high AFAR activity could be a means to control aflatoxin sensitivity in this poultry species.
Asunto(s)
Aflatoxina B1/análogos & derivados , Aldehído Reductasa/metabolismo , Aves de Corral/metabolismo , Aflatoxina B1/química , Aflatoxina B1/toxicidad , Animales , Femenino , Cinética , MasculinoRESUMEN
A study was conducted to determine the cytosolic in vitro hepatic enzymatic kinetic parameters Vmax, KM, and intrinsic clearance (CLint) for aflatoxin B1 (AFB1) reductase [aflatoxicol (AFL) production] and AFL dehydrogenase (AFB1 production) in four commercial poultry species (chicken, quail, turkey and duck). Large differences were found in AFB1 reductase activity, being the chicken the most efficient producer of AFL (highest CLint value). Oxidation of AFL to AFB1 showed only slight differences among the different poultry species. On average all species produced AFB1 from AFL at a similar rate, except for the turkey which produced AFB1 from AFL at a significantly lower rate than chickens and quail, but not ducks. Although the turkey and duck showed differences in AFL oxidation Vmax and KM parameters, their CLint values did not differ significantly. The ratio AFB1 reductase/AFL dehydrogenase enzyme activity was inversely related to the known in vivo sensitivity to AFB1 being highest for the chicken, lowest for the duck and intermediate for turkeys and quail. Since there is no evidence that AFL is a toxic metabolite of AFB1, these results suggest that AFL production is a detoxication reaction in poultry. Conversion of AFB1 to AFL prevents the formation of the AFB1-8,9-exo-epoxide which, upon conversion to AFB1-dihydrodiol, is considered to be the metabolite responsible for the acute toxic effects of AFB1.