Detoxification of Aflatoxin-Contaminated Maize by Neutral Electrolyzed Oxidizing Water

Aflatoxins, a group of extremely toxic mycotoxins produced by Aspergillus flavus, A. parasiticus and A. nomius, can occur as natural contaminants of certain agricultural commodities, particularly maize. These toxins have been shown to be hepatotoxic, carcinogenic, mutagenic and cause severe human and animal diseases. The effectiveness of neutral electrolyzed oxidizing water (NEW) on aflatoxin detoxification was investigated in HepG2 cells using several validation methodologies such as the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay, the induction of lipid peroxidation, the oxidative damage by means of glutathione modulation, the Ames test and the alkaline Comet assay. Our results showed that, after the aflatoxin-contaminated maize containing 360 ng/g was soaked in NEW (60 mg/L available chlorine, pH 7.01) during 15 min at room temperature, the aflatoxin content did not decrease as confirmed by the immunoaffinity column and ultra performance liquid chromatography methods. Aflatoxin fluorescence strength of detoxified samples was similar to untreated samples. However, aflatoxin-associated cytotoxicity and OPEN ACCESS Toxins 2015, 7 4295 genotoxicity effects were markedly reduced upon treatment. According to these results, NEW can be effectively used to detoxify aflatoxin-contaminated maize

Actividad esporicida de la solución electrolizada con pH neutro en hongos de importancia poscosecha

The electroless water with neutral pH (7) is a novel antimicrobial agent, which has an effect on a large variety of microorganisms, safe for humans and the environment. The efficiency of electrolyzed super oxidation water with neutral pH (SES) in the reduction of spore germination and development of the germinative tube in fungi of postharvest importance was determined. A suspension of 8 × 10 7 mL -1 spores of the fungi Botrytis cinerea isolated from blackberry, Colletotrichum gloeosporioides isolated from mango, guava and lychee, Fusarium solani isolated from chili and stevia, Monilinia fructicola isolated from peach, Penicillium digitatum isolated from Mexican lemon and Persian lemon, Penicillium sp., papaya isolate and Rhizopus stolonifer isolated from yaca and soursop, were in contact with the SES for 5 min at concentrations of 3, 5, 6, 8, 18, 24, 27, 29, 36 and 43 ppm free chlorine, and sterile distilled water (control). Spores were seeded in culture medium potato dextrose agar (Bioxon ® ); the evaluations were carried out at 24 and 48 hours after sowing. The 100% inhibition of spore germination and Botrytis germ length was observed in the concentration range of 18-43 ppm, Colletotrichum (6-43 ppm), Fusarium (6-43 ppm), Monilinia (8 and 24-43 ppm), Penicillium (18-43 ppm), Rhizopus isolated from soursop (5-24 and 29-43 ppm) and Rhizopus of yaca (18-43 ppm). The results suggest that SES could be used as an alternative control of post-harvest fungi. Palabras clave: Disease, fruit, pathogen, rot, spores.El agua electrolizada con pH neutro (7) es un novedoso agente antimicrobiano, que tiene efecto en una gran variedad de microorganismos, seguro para los seres humanos y el medio ambiente. Se determinó la eficacia del agua electrolizada de súper oxidación con pH neutro (SES) en la reducción de la germinación de esporas y desarrollo del tubo germinativo en hongos de importancia postcosecha. Una suspensión de 8 × 10 7 esporas mL -1 de los hongos Botrytis cinerea aislado de zarzamora, Colletotrichum gloeosporioides aislado de mango, guayaba y lichi, Fusarium solani aislado de chile y estevia, Monilinia fructicola aislado de durazno, Penicillium digitatum aislado de limón mexicano y limón persa, Penicillium sp., aislado de papaya y Rhizopus stolonifer aislado de yaca y guanábana, estuvieron en contacto con la SES por 5 min a concentraciones de 3, 5, 6, 8, 18, 24, 27, 29, 36 y 43 ppm de cloro libre, y agua destilada estéril (testigo). Las esporas se sembraron en medio de cultivo papa dextrosa agar (Bioxon ® ); las evaluaciones se realizaron a las 24 y 48 horas posteriores a la siembra. La inhibición de 100% en la germinación de esporas y longitud del tubo germinativo de Botrytis se observó en el rango de concentración de 18-43 ppm, Colletotrichum (6-43 ppm), Fusarium (6 - 43 ppm), Monilinia (8 y 24-43 ppm), Penicillium (18-43 ppm), Rhizopus aislado de guanábana (5-24 y 29-43 ppm) y Rhizopus de yaca (18-43 ppm). Los resultados sugieren que la SES podría ser utilizada como alternativa de control de hongos poscosecha. Palabras clave: Enfermedad, esporas, fruto, pudrición, patógeno

A Theoretical Study of 8-Chloro-9-Hydroxy-Aflatoxin B1, the Conversion Product of Aflatoxin B1 by Neutral Electrolyzed Water

Theoretical studies of 8-chloro-9-hydroxy-aflatoxin B1 (2) were carried out by Density Functional Theory (DFT). This molecule is the reaction product of the treatment of aflatoxin B1 (1) with hypochlorous acid, from neutral electrolyzed water. Determination of the structural, electronic and spectroscopic properties of the reaction product allowed its theoretical characterization. In order to elucidate the formation process of 2, two reaction pathways were evaluated—the first one considering only ionic species (Cl+ and OH−) and the second one taking into account the entire hypochlorous acid molecule (HOCl). Both pathways were studied theoretically in gas and solution phases. In the first suggested pathway, the reaction involves the addition of chlorenium ion to 1 forming a non-classic carbocation assisted by anchimeric effect of the nearest aromatic system, and then a nucleophilic attack to the intermediate by the hydroxide ion. In the second studied pathway, as a first step, the attack of the double bond from the furanic moiety of 1 to the hypochlorous acid is considered, accomplishing the same non-classical carbocation, and again in the second step, a nucleophilic attack by the hydroxide ion. In order to validate both reaction pathways, the atomic charges, the highest occupied molecular orbital and the lowest unoccupied molecular orbital were obtained for both substrate and product. The corresponding data imply that the C9 atom is the more suitable site of the substrate to interact with the hydroxide ion. It was demonstrated by theoretical calculations that a vicinal and anti chlorohydrin is produced in the terminal furan ring. Data of the studied compound indicate an important reduction in the cytotoxic and genotoxic potential of the target molecule, as demonstrated previously by our research group using different in vitro assays

Ameliorative Effects of Neutral Electrolyzed Water on Growth Performance, Biochemical Constituents, and Histopathological Changes in Turkey Poults during Aflatoxicosis

Different in vitro and in silico approaches from our research group have demonstrated that neutral electrolyzed water (NEW) can be used to detoxify aflatoxins. The objective of this investigation was to evaluate the ability of NEW to detoxify B-aflatoxins (AFB1 and AFB2) in contaminated maize and to confirm detoxification in an in vivo experimental model. Batches of aflatoxin-contaminated maize were detoxified with NEW and mixed in commercial feed. A total of 240 6-day-old female large white Nicholas-700 turkey poults were randomly divided into four treatments of six replicates each (10 turkeys per replicate), which were fed ad libitum for two weeks with the following dietary treatments: (1) control feed containing aflatoxin-free maize (CONTROL); (2) feed containing the aflatoxin-contaminated maize (AF); (3) feed containing the aflatoxin-contaminated maize detoxified with NEW (AF + NEW); and (4) control feed containing aflatoxin-free maize treated with NEW (NEW). Compared to the control groups, turkey poults of the AF group significantly reduced body weight gain and increased feed conversion ratio and mortality rate; whereas turkey poults of the AF + NEW group did not present significant differences on productive parameters. In addition, alterations in serum biochemical constituents, enzyme activities, relative organ weight, gross morphological changes and histopathological studies were significantly mitigated by the aflatoxin-detoxification procedure. From these results, it is concluded that the treatment of aflatoxin-contaminated maize with NEW provided reasonable protection against the effects caused by aflatoxins in young turkey poults