Carlos A. A. de Almeida

Influence of package, type of apple juice and temperature on the production of patulin by Byssochlamys nivea and Byssochlamys fulva.

Although the production of patulin in apple fruits is mainly by Penicillium expansum, there is no information on the ability of heat resistant moulds that may survive pasteurization to produce this mycotoxin in juice packages during storage and distribution. In this study, the production of patulin by Byssochlamys spp (Byssochlamys nivea FRR 4421, B. nivea ATCC 24008 and Byssochlamys fulva IOC 4518) in cloudy and clarified apple juices packaged in laminated paperboard packages or in polyethylene terephthalate bottles (PET) and stored at both 21 degrees C and 30 degrees C, was investigated. The three Byssochlamys strains were able to produce patulin in both cloudy and clarified apple juices. Overall, the lower the storage temperature, the lower the patulin levels and mycelium dry weight in the apple juices (p<0.05). The greatest variations in pH and degrees Brix were observed in the juices from which the greatest mycelium dry weights were recovered. The maximum levels of patulin recovered from the juices were ca. 150 microg/kg at 21 degrees C and 220 microg/kg at 30 degrees C. HPLC-UV, HPCL-DAD and mass spectrometry analyses confirmed the ability of B. fulva IOC 4518 to produce patulin. Due to the heat resistance of B. nivea and B. fulva and their ability to produce patulin either in PET bottles or in laminated paperboard packages, the control of contamination and the incidence of these fungi should be a matter of concern for food safety. Control measures taken by juice industries must also focus on controlling the ascospores of heat resistant moulds.

Robotic automated clean-up for detection of fumonisins B1 and B2 in corn and corn-based feed by high-performance liquid chromatography

A high-performance liquid chromatography (HPLC) system with fluorescence detection and an automated on-line solid-phase extraction procedure for fumonisins B1 and B2 in corn and corn-based products is described. Different amounts of strong anion-exchange, C18 and end-capped C18 (C(18 ec)) silicas were tested for sample clean-up. Various HPLC parameters were analyzed. The best methodology was found to be extraction with acetonitrile-water and clean up on C(18 ec) disposable extraction cartridges. The system has the advantage of running in an unattended mode of operation and allows processing of 40 samples without system refuel, performing clean-up, o-phthaldialdehyde derivatization, injection and fumonisin detection by fluorescence detection linked to a computer integrator for automated data processing. Recoveries were performed with corn and corn-based feed samples (n=3) spiked with 0.1, 0.5, 1.0, 5.0 and 10 microg/g. Average recoveries for corn and corn-based feed were, respectively, 92.6 and 88.3% with relative standard deviations (RSDs) of 5.04 and 6.22%, for fumonisin B1 and 91.2 and 89.0% with RSDs of 5.84 and 7.88% for fumonisin B2. Detection limits (S/N=3) for corn and corn-based feed were approximately 0.03 microg/g for fumonisin B1 and 0.05 microg/g for fumonisin B2

Production of fumonisins by strains of Fusarium moniliforme according to temperature, moisture and growth period

Producao de fumonisinas B1 (FB1) e B2 (FB2) a partir de duas cepas brasileiras (LAMIC 2999/96 e 113F) e uma cepa americana (NRRL 13616) de Fusarium moniliforme foi avaliada em culturas de laboratorio submetidas a diferentes temperaturas (20, 25 e 30oC) e a diferentes teores de umidade (25, 34 e 42%) em substrato de milho. As culturas foram realizadas em periodos de 10, 20, 30, 45 e 60 dias, totalizando135 tratamentos com duas repeticoes para cada um. As fumonisinas foram extraidas com acetonitrila/agua. A limpeza foi realizada empregando cartuchos de silica C18 encapada (C18ec) e a derivacao com o-ftalodialdeido foram realizadas por um sistema processador automatico de amostras (ASPEC), seguidas por quantificacao das toxinas por CLAE. A producao de fumonisinas variou muito, atingindo rendimentos medios de 0,25 a 5515,45 µg/g de FB1 e de 0,15 a 3032,10 µg/g de FB2. Neste trabalho, os fatores como cepa, temperatura, umidade e dias de cultura fungica foram avaliados, e todos estes influenciaram nas quantidades de fumonisinas produzidas. As mais altas producoes de FB1 foram obtidas pela cepa 113F nas seguintes condicoes: teor de umidade de 34%, 60 dias de cultura, temperatura de 25oC. A maior producao media de FB2 foi obtida pela mesma cepa com culturas durante 45 dias, a um teor de umidade de 42%, a temperatura de 25oC. A temperatura ideal para producao de fumonisinas foi calculada por meio de analise de regressao, sendo 24,5oC e 24,3oC (±2oC) para FB1 e FB2, respectivamente.

Desempenho e plumagem de frangos de corte intoxicados por aflatoxinas

This work had the goal to study the exposure effect of broiler chickens to aflatoxins during a period of 42 days related to its development in size and feathers. Two experimental groups of male chicks were treated with ad libitum food, one group without aflatoxins (control group) and other using contaminated food with 3mg/kg of aflatoxins (treated group). Birds were evaluated twice a day to observe its performance and consumption of food. The weights were measured at the end of each breeding phase (21, 35 and 42 days). At the end of the experiment animal size, weight, amount of feathers, organs and amino acids profile in blood were analyzed and compared between the two groups. The treated group showed classical signs of intoxication with aflatoxins such as less uniformity, less feed consumption and body weight gain. Also some macroscopic and microscopic organs and tissue changes were observed. Evaluation of heart and liver of the treated group showed an increase in their average relative weight. Feather mass in this group was significantly reduced (33.8%), but its amino acids concentration was not affected despite the low performance of the intoxicated animals.

Determination of anti-anxiety and anti-epileptic drugs in hospital effluent and a preliminary risk assessment

In this study, an analytical methodology was developed for the determination of psycho-active drugs in the treated effluent of the University Hospital at the Federal University of Santa Maria, RS - Brazil. Samples were collected from point A (Emergency) and point B (General effluent). The adopted methodology included a pre-concentration procedure involving the use of solid phase extraction and determination by liquid chromatography coupled to mass spectrometry. The limit of detection for bromazepam and lorazepam was 4.9 ± 1.0 ng L(-1) and, for carbamazepine, clonazepam and diazepam was 6.1 ± 1.5 ng L(-1). The limit of quantification was 30.0 ± 1.1 ng L(-1), for bromazepam, clonazepam and lorazepam; for carbamazepine was 50.0 ± 1.8 ng L(-1) and was 40.0 ± 1.0 ng L(-1) for diazepam. The mean concentrations in the Emergency and General effluent treated currents were as follows: for bromazepam, 195 ± 6 ng L(-1) and 137 ± 7 ng L(-1); for carbamazepine, 590 ± 6 ng L(-1) and 461 ± 10 ng L(-1); for diazepam, 645 ± 1 ng L(-1) and 571 ± 10 ng L(-1); for lorazepam, 96 ± 7 ng L(-1) and 42 ± 4 ng L(-1); and for clonazepam, 134 ± 10 ng L(-1) and 57 ± 10 ng L(-1). A preliminary risk assessment was conducted: carbamazepine and diazepam require considerable attention owing to their environmental toxicity. The occurrence of these psychoactive-drugs and the environmental risks that they pose demonstrated the need for a more efficient treatment system. As far we are aware, there have been no comparable studies to this on the hazards of hospital effluents in Brazil, and very few that have carried out a risk assessment of psycho-active drugs in hospital effluent in general.

Seasonal variability of the essential oil of Hesperozygis ringens (Benth.) Epling.

This study was developed to evaluate the effect of seasonality on the yield and chemical composition of the essential oil (EO) of Hesperozygis ringens (Benth.) Epling, a native species from the Brazilian Pampa. Leaves were collected from four specimens of a single population in each of the four seasons for a year and were extracted in triplicate by hydro-distillation for 2 hours. The yield of EO (% w/w) was calculated on fresh weight basis (FWB), and the 16 oil samples were analyzed by gas chromatography coupled to mass spectrometry (GC-MS) and gas chromatography with flame ionization detector (GC-FID). Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA) were used as statistical tools to evaluate differences in chemical composition. The highest yields were obtained in autumn, spring and summer (2.32-4.38%), while the lowest yields were detected in winter, ranging from 1.15 to 1.91%. Oxygenated monoterpenoids were the predominant class of chemical constituents in the EO obtained in all seasons, showing the highest contents in autumn and summer, and pulegone was identified as a major compound, whose contents varied between 54.13 and 81.17%. The EO samples were divided into three chemical groups by HCA and PCA and were assigned to the same group, except for the three samples gathered in winter. The results showed a seasonal influence on the yield and chemical composition of the EO.

determination of patulin in apple juiCe by HplC uSing a Simple and faSt Sample preparation metHod

The goal of this work was to develop a simple and rapid preparation method for pa- tulin analysis in apple juice without previous clean-up. This method combined sonication and liquid extraction techniques and was used for determination of patulin in 37 commercial apple juices available on the market in the South of Brazil. The method performance characteristics were determined using a sample obtained in a local market fortified at five concentration levels of patulin and done in triplicates. The coefficient of variation for repeatability at the fortifica- tion level of 20.70µg.L -1 was 3.53 % and the recovery 94.63 %, respectively. The correlation coefficient was 0.9996 and agrees with the requirements for a linear analytical method value. The detection limit was 0.21µg.L -1 and the quantification limit 0.70 µg.L -1 . Only three of the analyzed samples were upper the allowed level of 50.00 µg.L -1 recommended for the World