Micotoxinas em silagens de milho do sul do Brasil e metodologia analítica para aflatoxinas por espectroscopia de infravermelho proximo em milho

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias, Programa de Pós-Graduação em Ciência dos Alimentos, Florianópolis, 2013.Esse trabalho reporta o desenvolvimento de metodologia para análise rápida de aflatoxinas (AFLs) em milho (Zea mays L.) por espectrometria através de sua detecção em comprimento de onda na faixa do infravermelho próximo (S-NIR). Para tal, foram utilizadas 450 amostras de milho em grão (contendo uma faixa ampla de contaminação bem como amostras negativas como Controle) previamente analisadas por método imunoenzimático (ELISA). Através de leituras destas amostras no S-NIR, foi definida e quantificada a curva de calibração correspondente a faixa de AFLs (amostras contendo AFLs em diferentes concentrações) que representem contaminações em milho (= e > do limite máximo tolerável- MTL do Mercosul e Brasil: 20 e 50 µg/kg, respectivamente para alimentos para animais de produção). O Limite de quantificação (LOQ) do método referência utilizado para quantificar previamente a contaminação por AFLs foi de 5 µg/kg. A partir dos dados referência, foram obtidos espectros no S-NIR e desenvolvida curva de calibração através de modelos matemáticos aplicados para verificar a viabilidade da análise de AFLs por S-NIR. Dos 450 espectros coletados (?: 400 a 2500 nm), 24 espectros foram selecionados para o tratamento matemático e aplicada a equação de regressão mínimo quadrados parciais - PLS (partial least-square). A faixa de detecção estabelecida foi de 6,1 a 62,0 µg/kg. O valor de R² (coeficiente de determinação) para a curva de AFLs no milho foi de 0,9885, SECV (erro padrão de validação cruzada)de 7,4508 e SEC (erro padrão de calibração) de 2,0584. Novos espectros de amostras contaminadas 92 deverão ser coletadas com objetivo de tornar a curva mais robusta bem como estabelecer seus respectivos limite de detecção (LOD) e LOQ.Abstract : This work reports the development of a methodology for rapid analysis of aflatoxins (AFLs) in maize (Zea mays L.) spectrometry through its detection wavelength in the near-infrared (NIR-S). To this end, we used 450 samples of corn (containing a wide range of contamination as well as negative control samples) previously analyzed by enzyme-linked immunosorbent assay (ELISA). Through readings of these samples in the NIR-S, was defined and quantified the calibration curve corresponding to band AFLs (AFLs samples containing different concentrations) that represent contamination in maize (= and> the maximum limit tolerable-LMT Mercosul and Brazil: 20 and 50 µg/kg, respectively for animal feed production). The limit of quantitation (LOQ) of the reference method used to quantify previously contamination AFLs was 5 µg/kg. From the data reference spectra were obtained on S-NIR calibration curve developed using mathematical models applied to verify the feasibility of analysis of AFLs by S-NIR. Of the 450 collected spectra (?: 400 to 2500 nm), 24 spectra were selected for the mathematical treatment and applied the regression equation partial least squares - PLS (partial least-square). The detection range was set from 6.1 to 62.0 µg/kg. The value of R² (coefficient of determination) for the curve AFLs maize was 0.9885, SECV (standard error of cross validation) of 7.4508 and SEC (standard error of calibration) of 2. New spectra contaminated samples should be collected in order to make the curve more robust as well as establish their respective LOD and LOQ

Productivity and the presence of mycotoxins in oats, wheat, and triticale subjected to grazing

This study aimed to evaluate the effects of grazing on the agronomic characteristics, grain yield, and presence of the mycotoxins aflatoxins, fumonisins, and zearalenone in IPR 126 oat, BRS Tarumã wheat, and IPR 111 triticale. This study was performed from April 24, 2012 to November 11, 2013 at UNIOESTE’s Experimental Farm in southern Brazil. The experimental design was a randomized block in a banded scheme, with four replications. The treatments in the A tracks were the three crops and the B tracks, the treatments: without grazing, one grazing event, or two grazing events. One grazing reduced the productivity of oat by 42.48% and triticale by 28.09% and increased wheat productivity by 24.89%. Two grazings reduced oat productivity by 54.14%, wheat by 40.96%, and triticale by 54.69%. Crops subjected to two grazings should be used for dry matter production for ground cover or partially used for silage when the plant is in the pasty or semi-hard grain stage, as grazing affects the agronomic characteristics and grain yield. The BRS Tarumã wheat, when used in crop-livestock integration systems, should be grazed only once due to increased productivity, while IPR 126 oat and IPR 111 triticale should not be grazed in crop-livestock integration systems for commercial production of grain. Oats from grazed plants should not be added to feeds of poultry in the early stage or pigs in the early and growth stages due to the higher levels of aflatoxins, fumonisins, and zearalenone. Grazing of winter crops increases the length of the crop cycle and grain formation will occur under climatic conditions favorable to the mycotoxins. Therefore, when grains are intended for human consumption, grazing is not recommended, as it increases the possibility of aflatoxins, fumonisins, and zearalenone at levels above the maximum tolerable levels permitted by law. These grains should be supplied to animals only after analysis of the mycotoxin levels present in the grains

Bromatological and mycotoxin analysis on soybean meal before and after the industrial process of micronization

Aflatoxins, fumonisins and zearalenone take part of the most studied mycotoxin groups due to their toxic effects on animal and human health. This research evaluated samples of soybeans meal used in animal food industry. A hundred and twenty one soybean meal samples were analyzed, so that 66 were analyzed before the industrial processing of micronization and 55 after it. The bromatological average of samples before micronization showed the following answers: 12.4% moisture; 46.4% protein; 79.5% protein solubility; 5.9% ash content; 2.2% fat; 4.3% fiber and 0.02 (ΔpH) of urease activity. The samples of micronization soybean meal showed 7.0% average values for moisture and 48.6% for crude protein. The mycotoxin levels were low in natura soybean meal; therefore, average values were 0.5μg kg-1, 29.6μg kg-1 and 56.8μg kg-1 for aflatoxin, zearelenone and fumonisin, respectively. After micronization, the average values for the studied samples were 1.3μg kg-1, 67.5μg kg-1 and 89.1μg kg-1, respectively for the same mycotoxins. The results for bromatological and mycotoxin analyses indicate similarity with the established patterns according to the Brazilian Compendium for Animal feed and reference literature. However, at least one of the three studied mycotoxin was detected in all of the analyzed samples and there was greater contamination of soybeans meal after the micronization process

Dehydration curve, fungi and mycotoxins in Tifton 85 hay dehydrated in the field and in shed

The objective of this study was to estimate the dehydration curve and occurrence of fungi and mycotoxins in Tifton 85 hay. The experimental design was randomized blocks in split plots with four replications and five levels of nitrogen (0, 25, 50, 75 and 100 kg ha-1) in the form of urea, two regrowth ages (28 and 35 days) and drying hay in the sun and in shed. After harvesting, the grass was sampled four times (0, 8, 23 and 32 hours) to dry in the sun and twelve times (0, 3, 18, 27, 42, 51, 66, 75, 90, 99, 114 and 123 hours) for drying in shed, turned over daily. The fungi were identified after seeding and growth in three steps of haymaking (cutting, baling and after 30 days of storage). To verify the presence of mycotoxins, 20 samples, composed of hay stored for 30 days, were collected, of which 10 samples were from sun-dried hay and 10 from hay dried in shed. It took 32 hours to produce hay at field conditions, with water loss rates up to 6.10 g g-1 DM-h during the first 8 hours after cutting. The average loss of water in this period was 2.0 g g-1 DM-h. Hays dried in the shed took 123 hours and the dehydration rates were less than 0.5 g g-1 DM-h due to environmental conditions. There was a predominance of three genera of fungi: Fusarium, Penicillium and Aspergillus. The largest population was the Fusarium, followed by Penicillium, at the steps of baling and storage. The fumonisin mycotoxin was found at a higher concentration and there was no difference in fumonisin concentration in the hay dried in the sun and in shed. Although it brings no hazard to animal health, the production of aflatoxin and zearalenone was significantly higher in the samples of sun-dried hay