Classification and compositional characterization of different varieties of cocoa beans by near infrared spectroscopy and multivariate statistical analyses.

Effective and fast methods are important for distinguishing cocoa varieties in the field and in the processing industry. This work proposes the application of NIR spectroscopy as a potential analytical method to classify different varieties and predict the chemical composition of cocoa. Chemical composition and colour features were determined by traditional methods and then related with the spectral information by partial least-squares regression. Several mathematical pre-processing methods including first and second derivatives, standard normal variate and multiplicative scatter correction were applied to study the influence of spectral variations. The results of chemical composition analysis and colourimetric measurements show significant differences between varieties. NIR spectra of samples exhibited characteristic profiles for each variety and principal component analysis showed different varieties in according to spectral features.

Antifungal effect and some properties of cell-free supernatants of two Bacillus subtilis isolates against Fusarium verticillioides.

Fusarium verticillioides causes significant decrease in corn yield and quality, and produces fumonisins, which represent a serious risk to human and animal health. Bacillus species can be an effective and environmentally friendly alternative for F. verticillioides biological control. In this study, some properties of cell-free supernatants (CFSs) of two Bacillus spp. identified as Bacillus subtilis (NT1, NT2) as well as the antifungal effect against F. verticillioides 97L were evaluated. B. subtilis NT1 and NT2 were isolated from commercially available fermented whole soybeans (Natto). Antifungal activity was observed in both CFSs of B. subtilis isolates (50-59 mm) obtained by co-culture suggesting that antifungal compound production depends on interaction between bacteria and fungi. Cell-free supernatants from the two B. subtilis isolates inhibited mycelial growth (77%-94%) and conidial germination (22%-74%) of F. verticillioides 97L. In addition, CFSs caused significant morphological changes such as distorted and collapsed hyphae with wrinkled surfaces and the presence of a large amount of extracellular material compared to the control without CFSs. Both B. subtilis isolates (NT1 and NT2) produced extracellular proteases, biosurfactants and polar low molecular weight compounds that probably act synergistically and may contribute to the antifungal activity. Antifungal compounds showed heat and pH stability and resistance to proteolytic enzymes. Furthermore, antifungal compounds showed high polarity, high affinity to water and a molecular weight less than 10 kDa. These results indicated that the two B. subtilis (NT1 and NT2) have potential as biocontrol agents for F. verticillioides.

Limosilactobacillus reuteri as sustainable biological control agent against toxigenic Fusarium verticillioides.

Corn contamination with Fusarium verticillioides (Sacc.) Nirenberg is a worldwide problem that affects yield and grain quality resulting in severe economic losses and implications for food safety. Control of F. verticillioides is a challenge, but lactic acid bacteria (LAB) has high potential as a biological control agent. In this study, the antifungal effect of Limosilactobacillus reuteri (formerly Lactobacillus reuteri) LR-92 against F. verticillioides 97L was investigated. Cell-free supernatant (CFS) from L. reuteri showed concentration-dependent fungicidal and fungistatic activity against F. verticillioides 97L. The antifungal compounds from CFS showed heat stability and pH dependence, and antifungal activity was not affected by treatment with proteolytic enzymes. High-performance liquid chromatography analysis indicated that L. reuteri LR-92 produces lactic and acetic acids. After liquid-liquid extraction, electrospray ionization mass spectrometry analysis of the active ethyl acetate fraction containing antifungal compounds revealed the production of 3-phenyllactic acid, cyclo-(L-Pro-L-Leu), cyclo-(L-Pro-L-Phe), and cyclo-(L-Phe-trans-4-OH-L-Pro). L. reuteri LR-92 has potential as a biocontrol agent for F. verticillioides and contributes to food safety.

Microcystin-Detoxifying Recombinant Saccharomyces cerevisiae Expressing the mlrA Gene from Sphingosinicella microcystinivorans B9.

Contamination of water by microcystins is a global problem. These potent hepatotoxins demand constant monitoring and control methods in potable water. Promising approaches to reduce contamination risks have focused on natural microcystin biodegradation led by enzymes encoded by the mlrABCD genes. The first enzyme of this system (mlrA) linearizes microcystin structure, reducing toxicity and stability. Heterologous expression of mlrA in different microorganisms may enhance its production and activity, promote additional knowledge on the enzyme, and support feasible applications. In this context, we intended to express the mlrA gene from Sphingosinicella microcystinivorans B9 in an industrial Saccharomyces cerevisiae strain as an innovative biological alternative to degrade microcystins. The mlrA gene was codon-optimized for expression in yeast, and either expressed from a plasmid or through chromosomal integration at the URA3 locus. Recombinant and wild yeasts were cultivated in medium contaminated with microcystins, and the toxin content was analyzed during growth. Whereas no difference in microcystins content was observed in cultivation with the chromosomally integrated strain, the yeast strain hosting the mlrA expression plasmid reduced 83% of toxins within 120 h of cultivation. Our results show microcystinase A expressed by industrial yeast strains as a viable option for practical applications in water treatment.

Pesticides removal from water using activated carbons and carbon nanotubes.

The conventional water treatment technique (CT) widely applied cannot alone remove pesticides efficiently from water. Therefore, this work aimed to provide technical and scientific support for the association of pulverized activated carbon (PACs), granular activated carbon (GACs), and carbon nanotubes (CNT) with CT concerning atrazine (ATZ), simazine (SMZ), and diuron (DIU) removal. Actual conditions of pre/during, and post-treatment points of application, within water production process line, in water treatment plants (WTPs), using the pesticides in two forms, commercial product (CP) and analytical standard (SD). It was possible to demonstrate significant differences regarding the removal of ATZ, SMZ, and DIU in their SD and CP forms for the PACs, GACs, and CNTs. The minimum dosage of CNT required for adequate adsorption of all pesticides was superior to 160 mg. L-1; is 400% higher than the minimum dosage of 40 mg. L-1 is required for PAC application. ATZ, SMZ, and DIU in the SD form were more efficiently removed with percentages superior to 96.4% for ATZ, 98.2% for SMZ, and 99.1% for DIU. The characteristics of the adsorptive materials did not guide the adsorption efficacy. Instead, chemical interaction, contact time, and point application were critical factors. The pre-treatment and post-treatment applications were the most efficient, with removals oscillating from 97.7% to 100% for ATZ, 97.7% to 100% for SMZ, and 99.1 to 100% for DIU PAC and GAC, respectively.

Fusarium verticillioides: evaluation of fumonisin production and effect of fungicides on in vitro inhibition of mycelial growth.

The objectives of this study were to evaluate the fumonisin production by 16 F. verticillioides strains on corn cultures and the effect of quintozene and fludioxonil + metalaxyl-M fungicides on "in vitro" mycelial growth on agar. In addition, the effect of fludioxonil + metalaxyl-M on fumonisin production in defined liquid culture medium was analyzed. Fumonisin B(1) levels on corn cultures ranged from 2.41 to 3996.36 µg/g and the F. verticillioides 103F strain produced the highest level (3996.36 ± 390.49 µg/g, P < 0.05). F. verticillioides strains were inoculated in potato dextrose agar with the addition of quintozene (75 to 9,375 µg/ml) and fludioxonil + metalaxyl-M (1.5 + 0.6 to 187.5 + 75 µg/ml) in order to evaluate the effect of these fungicides on "in vitro" mycelial growth. The F. verticillioides strains showed great variability concerning ED(50) values, which were below the recommended application dose for quintozene, but above that for fludioxonil + metalaxyl-M. Moreover, fungicide addition to the culture medium increased mean FB(1) levels compared to the control, suggesting the importance of focusing on the effect of fungicides on mycotoxin production as well as on the phytopathogen control.

Tryptophan Attenuates the Effects of OTA on Intestinal Morphology and Local IgA/IgY Production in Broiler Chicks.

Ochratoxin A (OTA) is a mycotoxin produced by species of Penicillium and Aspergillus that can contaminate products of plant origin that are used as animal feed. Through oral exposure, this mycotoxin primarily affects the chicken gastrointestinal system. The present study evaluated the intestinal toxic effects of OTA and the introduction of L-tryptophan to alleviate these effects in chickens. One-day-old chicks were exposed to a single OTA dose (1.4 mg/kg body weight-b.w.) and treated with or without four daily doses of L-tryptophan (100 mg/kg b.w.). Duodenal villus height/crypt depth, fecal immunoglobulin A/immunoglobulin Y (IgA/IgY) levels, and duodenal positive immunoglobulin A cells (IgA+) were evaluated by histology, ELISA, and immunohistochemistry, respectively, on the 14th day. There were significant changes in the duodenal villus height, crypt depth, and levels of fecal IgA/IgY and duodenal IgA+ cells (p < 0.05) in groups exposed to OTA. On the other hand, groups exposed to OTA and treated with L-tryptophan showed similar levels of villus height, IgA/IgY levels, and duodenal IgA+ cells to those of the control group (p > 0.05). In conclusion, exposure to a single dose of OTA orally induces changes in intestinal morphology, levels of IgA/IgY antibodies, and IgA+ cells. Thus, treatment with L-tryptophan may be a valid alternative means to reduce the harmful effects of OTA on the intestinal mucosa, which requires further study.

Further investigation of microbial degradation of microcystin using the advanced Marfey method.

It is known that microcystin (MC) is subject to microbial degradation to provide three types of products, linearized MCLR (Adda-Glu-Mdha-Ala-Leu-MeAsp-Arg), tetrapeptide Adda-Glu-Mdha-Ala, and Adda. They can be readily detected by the usual HPLC, because they commonly have an Adda moiety with a diene and an absorption maximum at 238 nm as the chromophore. However, no other degradation products without such a chromophore have been isolated to date. In this study, cell preparation of a bacterium B-9 that can degrade MC and detection of the degradation products were devised. First, we regulated the B-9 hydrolytic activity by washing with sodium chloride solution to obtain a desired cell preparation, which permitted an additional intermediate and the final products of MCLR to be obtained. Second, the resulting products could be firmly identified using the advanced Marfey method with the aid of log D. As a result of these experiments, the following degradation products were further identified: a tetrapeptide, Adda-Glu-Mdha-Ala, tripeptides Adda-Glu-Mdha, Glu-Mdha-Ala, and Arg-MeAsp-Leu, a dipeptide, Glu-Mdha, and amino acids Adda, Arg, and methylamine derived from Mdha. The present study expands the hydrolytic activity of the B-9 strain, which can hydrolyze not only cyanobacterial cyclic peptides but also MC to the intermediates and final products. The established characterization method composed of the advanced Marfey method and log D would be a standard technique for the structural characterization of a mixture of amino acids and peptides.

Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay to Detect Fusarium verticillioides in Poultry Feed Samples.

Fumonisins are a group of toxic secondary metabolites that are produced by Fusarium verticillioides which are associated with poultry health hazard and great economic losses. The objective of the present study was to develop an immunological method to detect F. verticillioides in poultry feed samples. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a polyclonal antibody against 67 kDa protein of the F. verticillioides 97K exoantigen was developed to detect this fungus. Antibody anti-67 kDa protein showed cross-reactivity against F. graminearum (2⁻7%) and F. sporotrichioides (10%), but no or low cross-reactivity against Aspergillus sp. and Penicillium sp. exoantigens. The detection limit for the 67 kDa protein of F. verticillioides was 29 ng/mL. Eighty-one poultry feed samples were analyzed for Fusarium sp. count, 67 kDa protein of F. verticillioides and fumonisin concentrations. Eighty of the 81 feed samples (98.6%) showed Fusarium sp. contamination (mean 6.2 x 104 CFU/g). Mean 67 kDa protein and fumonisin concentration in the poultry feed samples was 21.0 µg/g and 1.02 µg/g, respectively. The concentration of 67 kDa protein, as determined by ic-ELISA correlated positively (p < 0.05) with fumonisin levels (r = 0.76). These results suggest that this ic-ELISA has potential to detect F. verticillioides and predict fumonisin contamination in poultry feed samples.

Effect of Zingiber officinale Roscoe essential oil in fungus control and deoxynivalenol production of Fusarium graminearum Schwabe in vitro.

Members of the Fusarium genus are capable of contaminating agricultural commodities, compromising the quality of maize and other grains, which leads to severe quality and yield losses. Contamination with mycotoxins is also a concern. Essential oils are possible alternatives to the use of synthetic pesticides for control of fungal contamination, as many have antifungal and anti-mycotoxigenic properties and are innocuous to human health. They also do not cause any sort of microbial resistance and do not promote environmental pollution. The aim of this study was to evaluate the antifungal and anti-mycotoxigenic effects of Zingiber officinale Roscoe essential oil (GEO) upon Fusarium graminearum Schwabe in vitro. The essential oil was extracted by hydrodistillation and analysed by GC/MS. Antifungal and anti-mycotoxigenic activities were assessed by HPLC/UV by quantifying ergosterol and deoxynivalenol (DON), respectively. Results indicated that GEO inhibited ergosterol production at a concentration of 1000 µg/mL and DON production at a concentration of 500 µg/mL, evidencing that the anti-mycotoxigenic effect is independent of the antifungal effect due to its probable direct action upon toxin biosynthesis.

Aflatoxins and ochratoxin A in different cocoa clones (Theobroma cacao L.) developed in the southern region of Bahia, Brazil.

Brazil is the sixth largest producer of cocoa beans in the world, after Côte d'Ivoire, Ghana, Indonesia, Nigeria and Cameroon. The southern region of Bahia stands out as the country's largest producer, accounting for approximately 60% of production. Due to damage caused by infestation of the cocoa crop with the fungus Moniliophthora perniciosa, which causes 'witch's broom disease', research in cocoa beans has led to the cloning of species that are resistant to the disease; however, there is little information about the development of other fungal genera in these clones, such as Aspergillus, which do not represent a phytopathogenicity problem but can grow during the pre-processing of cocoa beans and produce mycotoxins. Thus, the aim of this work was to determine the presence of aflatoxin (AF) and ochratoxin A (OTA) in cocoa clones developed in Brazil. Aflatoxin and ochratoxin A contamination were determined in 130 samples from 13 cocoa clones grown in the south of Bahia by ultra-performance liquid chromatography with a fluorescence detector. The method was evaluated for limit of detection (LOD) (0.05-0.90 µg kg-1), limit of quantification (0.10-2.50 µg kg-1) and recovery (RSD) (89.40-95.80%) for AFB1, AFB2, AFG1, AFG2 and OTA. Aflatoxin contamination was detected in 38% of the samples in the range of

Dehydroabietic acid isolated from Pinus elliottii exerts in vitro antileishmanial action by pro-oxidant effect, inducing ROS production in promastigote and downregulating Nrf2/ferritin expression in amastigote forms of Leishmania amazonensis.

Dehydroabietic acid (DHA) is one of the main constituents of the resin that have antiprotozoal activity against Leishmania spp., but the leishmanicidal mechanism is unknown. The objective of the study was to investigate in vitro the leishmanicidal activity of the natural compound DHA against intracellular and extracellular forms of L. amazonensis and the mechanism of action involved. The antileishmanial activity of DHA was evaluated in vitro against promastigote forms of L. amazonensis by counting in Neubauer chamber. The morphological changes were observed by scanning electron microscopy and cell death mechanism by fluorescence assay using 2',7'-dichlorofluorescein diacetate probe (H2DCFDA), tetramethylrhodamine ethyl ester (TMRE), annexin-V and propidium iodide (PI). The antiamastigote effect was observed by counting the number of amastigotes per macrophage and percentage of infected cells. In addition, reactive oxygen species (ROS) production, nitric oxide (NO), cytokines, free iron and total iron-binding capacity (TIBC), expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and ferritin were evaluated. DHA inhibited the proliferation of promastigotes at all times tested. The compound (IC50, 40 ±â€¯0.1458 µg/mL) altered the morphology of the promastigote forms, caused mitochondrial depolarization, induced ROS production, increased phosphatidylserine exposure and caused loss of plasma membrane integrity. DHA also reduced the number of amastigotes and the percentage of infected macrophages by increasing ROS production, free iron and TIBC, and also caused downregulation of Nrf2 and ferritin expression. DHA was effective in the elimination of L. amazonensis both in its promastigote forms by apoptosis-like mechanisms and intracellular form by ROS production.

Impact of industrial dry-milling on fumonisin redistribution in non-transgenic corn in Brazil.

The aim of this study was to evaluate the fate of fumonisins B1 (FB1) and B2 (FB2) during industrial dry-milling in two lots from 2014 (n=120) and 2015 (n=120) of non-transgenic corn and their fractions (germ, pericarp, endosperm, cornmeal and grits), collected from one of the major Brazilian milling industries. Fumonisins were concentrated in the germ and pericarp at a rate of 322% and 188% (lot 1) and 311% and 263% (lot 2), respectively. In the endosperm, cornmeal and grits fumonisin levels decreased from 60 to 95%. Fumonisin levels in cornmeal and grits were below the maximum limit tolerated by the European Commission. Therefore, corn industrial dry-milling can contribute to reducing fumonisin levels in corn products intended for human consumption.

Impact of a Single Oral Acute Dose of Aflatoxin B1 on Liver Function/Cytokines and the Lymphoproliferative Response in C57Bl/6 Mice.

Aflatoxin B1 (AFB1), a mycotoxin found in food and feed, exerts harmful effects on humans and animals. The liver is the earliest target of AFB1, and its effects have been evaluated in animal models exposed to acute or chronic doses. Considering the possibility of sporadic ingestion of AFB1-contaminated food, this study investigated the impact of a single oral dose of AFB1 on liver function/cytokines and the lymphoproliferative response in mice. C57BL/6 mice were treated with a single oral AFB1 dose (44, 442 or 663 µg AFB1/kg of body weight) on the first day. Liver function (ALT, γ-GT, and total protein), cytokines (IL-4, IFN-γ, and IL-17), histopathology, and the spleen lymphoproliferative response to mitogens were evaluated on the 5th day. Although AFB1 did not produce any significant changes in the biochemical parameters, 663 µg AFB1/kg-induced hepatic upregulation of IL-4 and IFN-γ, along with liver tissue injury and suppression of the lymphoproliferative response to ConA (p < 0.05). In conclusion, a single oral dose of AFB1 exposure can induce liver tissue lesions, liver cytokine modulation, and immune suppression in C57BL/6 mice.

VIS-NIR spectroscopy as a process analytical technology for compositional characterization of film biopolymers and correlation with their mechanical properties.

There is an increasing interest in the use of polysaccharides and proteins for the production of biodegradable films. Visible and near-infrared (VIS-NIR) spectroscopy is a reliable analytical tool for objective analyses of biological sample attributes. The objective is to investigate the potential of VIS-NIR spectroscopy as a process analytical technology for compositional characterization of biodegradable materials and correlation to their mechanical properties. Biofilms were produced by single-screw extrusion with different combinations of polybutylene adipate-co-terephthalate, whole oat flour, glycerol, magnesium stearate, and citric acid. Spectral data were recorded in the range of 400-2498nm at 2nm intervals. Partial least square regression was used to investigate the correlation between spectral information and mechanical properties. Results show that spectral information is influenced by the major constituent components, as they are clustered according to polybutylene adipate-co-terephthalate content. Results for regression models using the spectral information as predictor of tensile properties achieved satisfactory results, with coefficients of prediction (R(2)C) of 0.83, 0.88 and 0.92 (calibration models) for elongation, tensile strength, and Young's modulus, respectively. Results corroborate the correlation of NIR spectra with tensile properties, showing that NIR spectroscopy has potential as a rapid analytical technology for non-destructive assessment of the mechanical properties of the films.

Prediction of chicken quality attributes by near infrared spectroscopy.

In the present study, near-infrared (NIR) reflectance was tested as a potential technique to predict quality attributes of chicken breast (Pectoralis major). Spectra in the wavelengths between 400 and 2500nm were analysed using principal component analysis (PCA) and quality attributes were predicted using partial least-squares regression (PLSR). PCA performed on NIR dataset revealed the influence of muscle reflectance (L(∗)) influencing the spectra. PCA was not successful to completely discriminate between pale, soft and exudative (PSE) and pale-only muscles. High-quality PLSR were obtained for L(∗) and pH models predicted individually (R(2)CV of 0.91 and 0.81, and SECV of 1.99 and 0.07, respectively). Water-holding capacity was the most challenging attribute to determine (R(2)CV of 0.70 and SECV of 2.40%). Sample mincing and different spectra pre-treatments were not necessary to maximise the predictive performance of models. Results suggest that NIR spectroscopy can become useful tool for quality assessment of chicken meat.

Natural occurrence of deoxynivalenol in wheat from Paraná State, Brazil and estimated daily intake by wheat products.

The occurrence of deoxynivalenol (DON) was evaluated in 113 wheat samples from the northern and central/southwestern regions of Paraná State, Brazil during the 2008 and 2009 growing seasons, and this rate of occurrence was used to estimate the DON dietary exposure. The DON determination was carried out by an indirect competitive enzyme-linked immunosorbent assay. DON was detected in 66.4% samples at levels ranging from 206.3 to 4732.3 µg/kg (mean 1894.9 µg/kg). The estimated daily intake (EDI) of DON through bread and pasta was evaluated in the inhabitants of Londrina City in northern Paraná State, Brazil. The average intake of these inhabitants was 0.79 µg/kg body weight (b.w.) for bread and 0.35 µg/kg b.w. for pasta. The total EDI was 1.13 µg/kg, which is above the Provisional Tolerable Daily Maximum Intake (PTDMI) of 1 µg/kg b.w. To our knowledge, this is the first report on natural DON occurrence in wheat and DON dietary exposure estimation from Paraná, Brazil.

Desaguamento do lodo de estações de tratamento de água por leito de drenagem/secagem / Dewatering of sludge from water treatment plants using drainage/drying bed

Vários estudos têm mostrado a eficiência das técnicas naturais de desaguamento do lodo das estações de tratamento de água no Brasil. O objetivo deste estudo foi avaliar o sistema de desaguamento por leito de drenagem/secagem, com manta geotêxtil dos lodos de decantadores de duas diferentes estações de tratamento de água, em ensaios de laboratório e escala reduzida. Foram considerados os parâmetros técnicos de projeto: densidade da manta; taxa de aplicação dos sólidos; taxa de aplicação volumétrica e secagem em diferentes condições climáticas. Na fase de drenagem, entre 78 a 91% do volume total dos líquidos percolados apresentaram qualidade compatível com o padrão de enquadramento dos corpos de água doce classes I e II, segundo a Resolução 357/05 do Conama. Na fase de secagem, foi possível obter teores de sólidos totais entre 15,9 e 20,3%, cujos valores são da mesma ordem de grandeza que aqueles obtidos por desaguamentos mecânicos, mesmo em condições críticas de secagem.

Several studies have showed the efficiency of natural techniques to dewater the sludge from water treatment plants in Brazil. The objective of this study was to evaluate the dewatering system through drainage/drying with a geotextile blanket using the sludge of decanters from two types of water treatment plants, in laboratory experiments and reduced scale. The following technical parameters of project were considered: density of blanket, application rate of solids, volumetric application rate, and drying in different climate conditions. In the drainage phase, 78 to 91% of the total volume of percolated liquid presented a compatible quality to the standardized limits for freshwater bodies from classes I and II, according to Resolution 357/05 from the National Council of Environment. In the drying phase, it was possible to obtain total solid rates between 15.9 and 20.3%, values of the same magnitude order as those obtained by mechanical drainages, even in critical drying conditions.

Blackberry (Rubus spp.): influence of ripening and processing on levels of phenolic compounds and antioxidant activity of the 'Brazos' and 'Tupy' varieties grown in Brazil / Amora Preta (Rubus spp.): influência do estádio de maturação e do processamento nos teores de compostos fenólicos e atividade antioxidante das variedades 'Brazos' e 'Tupy' cultivadas no Brasil

Fruits from temperate and tropical climates which have high levels of antioxidant compounds are the source of numerous studies concerning the correlation with benefits to human health. The objectives of this study were to quantify the anthocyanins and phenolic compounds and also to measure the antioxidant activity (ferric reducing antioxidant power - FRAP) of blackberries from two varieties grown in southern Brazil ('Brazos' and 'Tupy') at three stages of ripening; unripe, semi-ripe, ripe and their products (pulp and fermented products). During fruit ripening it was observed that weight, size, diameter and sugars increase significantly and acidity decreased significantly. The anthocyanin content ranged from 4.19 (semi-ripe 'Tupy' variety) to 205.75mg 100g-1 (ripe 'Brazos' variety). The highest levels of phenolic compounds were observed for the unripe fruit of both varieties, while antioxidant activity showed no significant difference during the ripening stages. The studied pulp showed a high content of phenolic compounds (ten times higher than that found in the ripe fruits). The anthocyanin content and antioxidant activity did not show the same increase due to the degradation of anthocyanins caused by the heat treatment that was used. The alcoholic fermented beverage made from blackberries remained stable (total phenolic compounds and antioxidant activity) during two years of storage, but the in third year a significant reduction in antioxidant activity was observed. These results can be important for establishing the shelf life of this kind of product made with blackberry.

Algumas frutas de clima temperado e tropical, principalmente as do tipo "berries", têm como característica o acúmulo de compostos antioxidantes e são objeto de inúmeros estudos, haja vista a sua correlação com os benefícios para a saúde humana. Os objetivos deste trabalho foram quantificar as antocianinas, os compostos fenólicos e mensurar a capacidade antioxidante (ferric reducing antioxidant power - FRAP) de amoras-pretas de duas cultivares ('Brazos' e 'Tupy'), nos três estágios de maturação, considerados verde, semi-maduro e maduro, e dos seus produtos (polpas e fermentados). No processo de maturação da fruta, observou-se que o peso, tamanho, diâmetro e teor de açúcar aumentaram significativamente enquanto a acidez diminuiu. Os teores de antocianinas aumentaram de 4,19 (FSM cv. 'Tupy') (frutos semi-maduros) para 205,75mg 100g-1 (FM cv. 'Brazos') (frutos maduros). Os maiores teores de compostos fenólicos foram verificados para os frutos verdes (FV) das duas cultivares, enquanto a atividade antioxidante não apresentou diferença significativa nos estádios de maturação. A polpa estudada acumulou um elevado teor de compostos fenólicos (dez vezes superior ao encontrado nas frutas maduras). Os teores de antocianinas e de atividade antioxidante não apresentaram o mesmo perfil em função da degradação das antocianinas pelo processo térmico utilizado. O fermentado alcoólico de amora permaneceu estável (fenóis totais e atividade antioxidante) durante dois anos de estocagem, porém, no terceiro ano, foi observada uma redução significativa da atividade antioxidante. Estes resultados são importantes para o estabelecimento da vida-de-prateleira desses produtos derivados da amora.

Purification of Candida guilliermondii and Pichia ohmeri killer toxin as an active agent against Penicillium expansum.

An antifungal assay with cell-free culture supernatant of Pichia ohmeri 158 and Candida guilliermondii P3 was tested against Penicillium expansum strain #2 at 25 degrees C by measuring hyphal length and percentage conidia germination. C. guilliermondii was more effective against P. expansum conidia germination (58.15% inhibition), while P. ohmeri showed higher inhibition of mycelial growth (66.17%), indicating a probable mechanism associated with killer activity. This killer toxin (molecular mass <3 kDa) was partially purified by normal phase HPLC, using TSKgel Amide-80 analytical and preparative columns. Compared with crude extract, the killer toxin eluted from the post analytical column significantly inhibited P. expansum:% inhibition rose from 42.16 to 90.93% (C. guilliermondii) and 39.32 to 91.12% (P. ohmeri) (p < 0.05). The one-step purification process was adequate in isolating killer toxin from culture supernatant and also increased anti-Penicillium activity.