Gaseous Ozonation to Reduce Aflatoxins Levels and Microbial Contamination in Corn Grits.

Corn is one of the most cultivated cereals in Brazil. However, its grains are constantly exposed to contamination by mycotoxins. Corn grits are used by the food industry to produce a large variety of corn products such as canjiquinha, a cultural food easily purchased by the Brazilian consumer at low prices. Some studies have demonstrated high contamination of this product by aflatoxins (AFs), representing a potential risk of exposure due to such a contamination. In this study, the efficacy of gaseous ozonation was evaluated on the levels of aflatoxins and on the microbial contamination of corn grits. The application of gaseous ozone was tested in different combinations of exposure time, ozone concentration, and canjiquinha mass. After the ozonation treatment, samples were collected for aflatoxin and microbiological analyses. Aflatoxins were evaluated using a high-performance liquid chromatography with fluorescence detection (HPLC-FD) system using pre-column derivatization, and the microbiological analyses were carried out for toxin-producer fungi and mesophilic bacteria. After ozone detoxification, results showed reductions up to 57% in aflatoxin levels. Total fungal count was reduced around 3.0 cycles log CFU g-1 and total mesophilic counts were reduced to non-detectable levels. These results demonstrated that ozonation is an effective alternative for reducing aflatoxin and microbial contamination in products like canjiquinha, thereby improving food safety.

Antigenotoxic Effect of Piperine in Broiler Chickens Intoxicated with Aflatoxin B1.

Piperine is an abundant amide extracted from black pepper seeds which has been shown to have protective effects against cytotoxic and genotoxic carcinogenesis induced by certain chemical carcinogens and aflatoxin B1 (AFB1) in vitro. The aim of this work was to study, in vivo, the antigenotoxic potential of feed-added piperine on broiler chickens experimentally intoxicated with AFB1, using micronucleus and comet assays. The antigenotoxicity assessment of 9-day-old chicks was performed on a total of 60 chickens divided into four groups of 15 broilers each: (C) control, (P) 60 mg·piperine kg-1 feed, (A) 0.5 mg·AFB1·kg-1 body weight, (daily by oral route), and (P + A) co-treatment with piperine and AFB1. The experiment was conducted for 26 days. Chicks intoxicated with AFB1 showed significant genotoxic effects in the first 24 h post intoxication, and the effects remained in the other periods analyzed (48, 72, and 96 h and 26 days of treatment). The DNA damage in peripheral blood cells, the number of erythrocytes with micronuclei, and polychromatic-to-normochromatic erythrocyte ratio were significantly reduced or absent in the piperine/AFB1 group. No significant differences were observed between the group piperine/AFB1 and the control and piperine-alone groups. The addition 60 mg·kg-1 of piperine to the diet of the broiler chicks was safe, promoting beneficial effects in poultry health with respect to the toxic effects 0.5 mg·AFB1·kg-1 body weight.

Administração oral de piperina em frangos de corte / Piperine oral administration on broiler' chikens

O efeito tóxico da piperina, principal amida presente em diversas espécies de pimenta, foi avaliada em frangos de corte por meio da administração oral (0,00, 1,12, 2,25 e 4,5mg kg-1 peso vivo) por 14 dias consecutivos. Foram empregados 60 pintos machos (Cobb Avian 48) com sete dias de idade, distribuídos aleatoriamente em quatro grupos experimentais (n=15). Foram avaliados parâmetros como: ganho de peso, peso relativo do fígado e alterações hematológicas e anatomopatológicas. A administração oral de piperina não interferiu no ganho de peso ou no peso relativo do fígado, além de não promover alteração no tamanho e na coloração dos órgãos ou no aparecimento de lesões de parênquima e nas mucosas do órgão. Todavia, alterações histopatológicas dose-dependentes foram observadas. A piperina não foi capaz de alterar significativamente os parâmetros hematológicos analisados, com exceção do leucograma, em que as doses de 1,12mg e 2,25mg kg-1 promoveram aumento do número de heterófilos e do número total de leucócitos, respectivamente. Os resultados sugerem que a dose oral de 1,12mg de piperina por quilo não é tóxica para frangos de corte, sendo semelhante aos resultados obtidos para ratos e camundongos. Além disso, constatou-se a capacidade da piperina em aumentar o número total de leucócitos circulantes a partir da dose de 2,25mg kg-1 nessa espécie animal.

The toxic effect of piperine, the main amide compound of different pepper species, was evaluated on broiler chickens by oral administration at 0.00, 1.12, 2.25 and 4.50mg kg-1 concentrations for 14 days. Sixty seven days old male chicks (Cobb Avian 48) randomly allocated to four experimental groups (n=15) were used in this work. Parameters such as: body weight gain, liver relative weight, hematological and anatomopathological alterations were analyzed. The oral route administration did not interfere on weight gain or liver relative weight, as well as, any modification on size and organs' color and/or parenchyma/mucous membranes injuries were observed; however, liver histopathological changes were noticed in a dose-dependent manner. In addition, piperine did not alter hematological parameters, except for leukocytes counting, which at 1.12 and from 2.25mg kg-1 caused an increase of heterophils and in the total number of leukocytes, respectively. The results suggest that 1.12mg kg-1 of piperine orally administrated is not toxic for broiler chickens, as previously observed for rats and mice. Moreover, 2.25mg kg-1 of piperine seems to increase the total number of leukocytes for this animal specie.

Characterization of Aspergillus species based on fatty acid profiles.

Cellular fatty acid (FA) composition was utilized as a taxonomic tool to discriminate between different Aspergillus species. Several of the tested species had the same FA composition and different relative FA concentrations. The most important FAs were palmitic acid (C16:0), estearic acid (C18:0), oleic acid (C18:1) and linoleic acid (C18:2), which represented 95% of Aspergillus FAs. Multivariate data analysis demonstrated that FA analysis is a useful tool for differentiating species belonging to genus Aspergillus. All the species analyzed showed significantly FA acid profiles (p < 0.001). Furthermore, it will be possible to distinguish among Aspergillus spp. in the Flavi Section. FA composition can serve as a useful tool for the identification of filamentous fungi.

Mycoflora and fumonisin contamination in Brazilian corn from sowing to harvest.

The present study aimed to analyze the mycoflora and potential mycotoxin contamination of soil and corn samples collected at different plant maturity stages in Capão Bonito and Ribeirão Preto, two regions of the State of São Paulo, Brazil. In addition, the data obtained were correlated with the occurrence of wind-dispersed fungi and the predominant climatic conditions of the two regions studied. Corn mycoflora profiles showed that Fusarium verticillioides prevailed in 35% of the samples from Capão Bonito and in 49% of the samples from Ribeirão Preto. Examination of wind-dispersed fungi also revealed a high incidence of F. verticillioides. Soil mycoflora analyses showed that Penicilliumwas the most prevalent genus, although F. verticillioides was present in 55.5% of Capão Bonito's samples and in 26.7% of Ribeirão Preto's samples. With respect to water activity, the corn kernels most contaminated with F. verticillioides had water activity levels of 0.70-0.80. HPLC analysis of fumonisins revealed that 88.5% of Capão Bonito's kernels were contaminated with fumonisin B(1) (FB(1)) (0.09-10.87 microg/g) and 53.8% with fumonisin B(2) (FB(2)) (0.05-0.52 microg/g); Ribeirão Preto's kernels presented contamination levels of 93.5% for FB(1) (0.11-17.69 microg/g) and 61.3% for FB(2) (0.05-5.24 microg/g). No aflatoxins were detected by thin-layer chromatography in corn grains of either region. The concomitant occurrence of F. verticillioides and fumonisins in most of the field corn assayed demonstrates the importance of an effective control of cultivation throughout the plant maturity stages.