Madeira wine vinification process: analysis and control

The present study was done in collaboration with J. Faria e Filhos company, a Madeira wine producer, and its main goal was to fully characterize three wines produced during 2014 harvest and identify possible improving points in the winemaking process. The winemaking process was followed during 4 weeks, being registered the amounts of grapes received, the fermentation temperatures, the time at which fermentation was stopped and evolution of must densities until the fortification time. The characterization of musts and wines was done in terms of density, total and volatile acidity, alcohol content, pH, total of polyphenol, organic acids composition, sugars concentration and the volatile profile. Also, it was developed and validated an analytical methodology to quantify the volatile fatty acids, namely using SPME-GC-MS. Briefly, the following key features were obtained for the latter methodology: linearity (R2=0.999) e high sensitivity (LOD =0.026-0.068 mg/L), suitable precision (repeatability and reproducibility lower than 8,5%) and good recoveries (103,11-119,46%). The results reveal that fermentation temperatures should be controlled in a more strictly manner, in order to ensure a better balance in proportion of some volatile compounds, namely the esters and higher alcohols and to minimize the concentration of some volatiles, namely hexanoic, octanoic and decanoic acids, that when above their odours threshold are not positive for the wine aroma. Also, regarding the moment to stop the fermentation, it was verified that it can be introduced changes which can also be benefit to guarantee the tipicity of Madeira wine bouquet.

Improving photofermentative hydrogen production through metabolic engineering and DOE (Design of Experiments)

A l’heure actuelle, les biocarburants renouvelables et qui ne nuit pas à l'environnement sont à l'étude intensive en raison de l'augmentation des problèmes de santé et de la diminution des combustibles fossiles. H2 est l'un des candidats les plus prometteurs en raison de ses caractéristiques uniques, telles que la densité d'énergie élevée et la génération faible ou inexistante de polluants. Une façon attrayante pour produire la H2 est par les bactéries photosynthétiques qui peuvent capter l'énergie lumineuse pour actionner la production H2 avec leur système de nitrogénase. L'objectif principal de cette étude était d'améliorer le rendement de H2 des bactéries photosynthétiques pourpres non sulfureuses utilisant une combinaison de génie métabolique et le plan des expériences. Une hypothèse est que le rendement en H2 pourrait être améliorée par la redirection de flux de cycle du Calvin-Benson-Bassham envers du système de nitrogénase qui catalyse la réduction des protons en H2. Ainsi, un PRK, phosphoribulose kinase, mutant « knock-out » de Rhodobacter capsulatus JP91 a été créé. L’analyse de la croissance sur des différentes sources de carbone a montré que ce mutant ne peut croître qu’avec l’acétate, sans toutefois produire d' H2. Un mutant spontané, YL1, a été récupéré qui a retenu l'cbbP (codant pour PRK) mutation d'origine, mais qui avait acquis la capacité de se développer sur le glucose et produire H2. Une étude de la production H2 sous différents niveaux d'éclairage a montré que le rendement d’YL1 était de 20-40% supérieure à la souche type sauvage JP91. Cependant, il n'y avait pas d'amélioration notable du taux de production de H2. Une étude cinétique a montré que la croissance et la production d'hydrogène sont fortement liées avec des électrons à partir du glucose principalement dirigés vers la production de H2 et la formation de la biomasse. Sous des intensités lumineuses faibles à intermédiaires, la production d'acides organiques est importante, ce qui suggère une nouvelle amélioration additionnel du rendement H2 pourrait être possible grâce à l'optimisation des processus. Dans une série d'expériences associées, un autre mutant spontané, YL2, qui a un phénotype similaire à YL1, a été testé pour la croissance dans un milieu contenant de l'ammonium. Les résultats ont montré que YL2 ne peut croître que avec de l'acétate comme source de carbone, encore une fois, sans produire de H2. Une incubation prolongée dans les milieux qui ne supportent pas la croissance de YL2 a permis l'isolement de deux mutants spontanés secondaires intéressants, YL3 et YL4. L'analyse par empreint du pied Western a montré que les deux souches ont, dans une gamme de concentrations d'ammonium, l'expression constitutive de la nitrogénase. Les génomes d’YL2, YL3 et YL4 ont été séquencés afin de trouver les mutations responsables de ce phénomène. Fait intéressant, les mutations de nifA1 et nifA2 ont été trouvés dans les deux YL3 et YL4. Il est probable qu'un changement conformationnel de NifA modifie l'interaction protéine-protéine entre NifA et PII protéines (telles que GlnB ou GlnK), lui permettant d'échapper à la régulation par l'ammonium, et donc d'être capable d'activer la transcription de la nitrogénase en présence d'ammonium. On ignore comment le nitrogénase synthétisé est capable de maintenir son activité parce qu’en théorie, il devrait également être soumis à une régulation post-traductionnelle par ammonium. Une autre preuve pourrait être obtenue par l'étude du transcriptome d’YL3 et YL4. Une première étude sur la production d’ H2 par YL3 et YL4 ont montré qu'ils sont capables d’une beaucoup plus grande production d'hydrogène que JP91 en milieu d'ammonium, qui ouvre la porte pour les études futures avec ces souches en utilisant des déchets contenant de l'ammonium en tant que substrats. Enfin, le reformage biologique de l'éthanol à H2 avec la bactérie photosynthétique, Rhodopseudomonas palustris CGA009 a été examiné. La production d'éthanol avec fermentation utilisant des ressources renouvelables microbiennes a été traitée comme une technique mature. Cependant, la plupart des études du reformage de l'éthanol à H2 se sont concentrés sur le reformage chimique à la vapeur, ce qui nécessite généralement une haute charge énergetique et résultats dans les émissions de gaz toxiques. Ainsi le reformage biologique de l'éthanol à H2 avec des bactéries photosynthétiques, qui peuvent capturer la lumière pour répondre aux besoins énergétiques de cette réaction, semble d’être plus prometteuse. Une étude précédente a démontré la production d'hydrogène à partir d'éthanol, toutefois, le rendement ou la durée de cette réaction n'a pas été examiné. Une analyse RSM (méthode de surface de réponse) a été réalisée dans laquelle les concentrations de trois facteurs principaux, l'intensité lumineuse, de l'éthanol et du glutamate ont été variés. Nos résultats ont montré que près de 2 moles de H2 peuvent être obtenus à partir d'une mole d'éthanol, 33% de ce qui est théoriquement possible.

Comportamentos alimentares e saúde oral em voleibolistas

Introdução e objectivos: A saúde oral em atletas é parte essencial para a saúde geral, sendo um factor determinante para a qualidade de vida e desempenho desportivo. Logo, um bom estado de saúde só existirá se a cavidade oral se encontrar ausente de patologias. Os desequilíbrios nutricionais possuem efeitos sobre a cavidade oral, condicionando assim a qualidade de vida e desempenho do atleta. Desta forma, o excesso de ingestão de alguns alimentos podem ser factores de risco para a saúde, tendo conta em que, a etiologia da cárie dentária está relacionada com a ação de microorganismos orais que produzem ácidos orgânicos, a partir do metabolismo dos hidratos de carbono. O objectivo deste estudo foi avaliar a saúde oral bem como os hábitos alimentares e o uso de protetores bucais durante a prática desportiva. Participantes e Métodos: Foram observados 55 atletas de voleibol entre os 15 e 18 anos de ambos os géneros, do clube de voleibol Academia José Moreira e Leixões. Tratou-se de um estudo transversal, no qual foi realizado exame clínico intraoral (índice de cárie CPOd, indice de erosão dentária BEWE) e preenchimento de questinário, em que os indivíduos foram caracterizados em 5 componentes: dados sociodemográficos (idade, peso e estatura), dados sobre perceção de saúde, dados sobre comportamentos de saúde oral, dados sobre prática desportiva e dados sobre comportamentos alimentares (questionário semi-quantitativo de frequência alimentar). A análise estatística descritiva e inferencial dos dados recolhidos foi realizada com o auxílio do programa informático SPSS, versão 23.0. Resultados: Os hábitos de saúde oral não são os mais adequados e a percentagem de atletas que visita o médico dentista é elevada para “só quando tem dores” ou “ocasionalmente”. A média do CPOD geral foi de 4,22 ± 4,55. Não houve diferenças estatisticamente significativas (p>0,05) entre o CPOD geral e o IMC. Nenhum dos atletas usa protetor bucal durante a prática desportiva. Os alimentos mais consumidos foram a carne, fruta, leite, peixe e, biscoitos, bolos e bolachas; e os menos consumidos foram as bebidas alcoólicas, mel ou compotas e café. Conclusão: Os hábitos de higiene oral são um melhor indicador do que o IMC para a presença de cárie. Não há relação direta entre índice CPOD e IMC. Seria importante prestar mais informação sobre vantagens do uso de protetores bucais junto dos atletas e de treinadores bem como, esclarecer que a consistência e as propriedades sensoriais ligadas à textura e à consistência dos alimentos na superfície dentária interferem com a cárie.

Ácidos orgânicos e probiótico na redução da colonização de Salmonella enterica Heidelberg em frangos de corte

In the food industry, Salmonella sp. and other pathogens represent a major threat as potential causes of foodborne diseases. The Salmonella enterica serovar Heidelberg (SH) has a great importance to public health, being currently the most frequent serotype in the official control analyzes carried out in Brazil’s Southern. In this region are concentrated the largest poultry flocks, contributing over 60 % of Brazilian production of chicken meat. To help prevent and minimize infections by pathogenic bacteria, in addition to attend the standards of biosecurity in the field and good hygiene and sanitizing programs in the industry, there is the possibility to control directly on the poultry breeding. Use of organic acids added in the diet of the fowls has an antibacterial action. In the other hand, the use of probiotics has been shown to prevent enteric diseases. This dissertation was divided into two chapters and aimed to evaluate the quantitative and qualitative reduction of SH in broiler chickens. In the first chapter the aim was to collect information on the prevalence rate of SH strain in broiler chickens. This coefficient was 100 %, being a suitable method for determining the frequency of salmonellosis in broiler chickens. In the second chapter five treatments were evaluated (a positive control and four tests), in order to reduce the colonization of SH in broilers, using organic acids and probiotic during production phase. In SH count related to the colonization of the cecum at 28 days of age differences were observed (p<0,05) among treatments. The Treatment 2 showed a count of 41 most probable number of bacterial cells per gram (MPN/g) with lower counts when compared to the positive control (5.106 MPN/g) and treatment 1 (1.354 MPN/g). The evaluation index of infected birds, there was a difference between treatments. In treatment 2, and treatment 4 reduced 58,34 % and 25 % respectively the number of fowls infected with SH. There was no effect of treatments on the performance. Administration of organic acids and probiotic used in the treatment 2 proved to be an effective tool in the SH control in broilers at 28 days of age.

Atividade antimicrobiana de ácidos orgânicos e compostos clorados sobre micro-organismos patogênicos em carne de frango

The current scenario of the Brazilian poultry production is defined by high productivity motivated by exports to markets with elevated levels of sanitary requirement. The work aimed to evaluate the efficacy of chlorinated compounds (chlorine dioxide, dichloro and trichloro) and organic acids (citric, lactic and peracetic acids) in reducing the contamination of poultry by Salmonella spp., mesophiles and enterobacteriaceae. Were isolated 102 strains Salmonella spp. poultry carcass from June to September 2014. Strains were identified by PCR. Was determined the minimum inhibitory concentration (MIC) of antimicrobial compounds for the standard strains of S. Typhimurium, S. Enteritidis and S. Heidelberg. MIC of lactic acid and peracetic acid (20 to 10 g/L) was applied in strains of Salmonella spp. isolated from the slaughter. The MIC of the compounds lactic acid and sodium dichloro was applied in contaminated chiller water with Salmonella (109 CFU/mL) and this was determined Salmonella count in water. Thighs and drumsticks poultry were contaminated with S. Heidelberg (109 UFC/mL) and were applied dichloro (60 mg/L), lactic acid (20 g/L) and sodium hypochlorite (5,0 and 0,5 mg/L) compounds. In the identification by PCR, 93,1% of the strains were identified as Salmonella. For sodium dichloro the MIC was 60 mg/L for 15 minutes to S. Heidelberg and 60 mg/L for 20 minutes for S. Enteritidis. Lactic acid presented MIC of the 5 g/L for 10 minutes to S. Enteritidis 10 g/L for 15 minutes to S. Typhimurium and 20 g/L for 20 minutes to S. Heidelberg. For peracetic acid, MICs were 10 g/L for 10 minutes to S. Typhimurium and S. Heidelberg and 10 g/L for 20 minutes to S. Enteritidis. To citric acid, MICs were 10 g/L for 10 minutes to S. Typhimurium and S. Enteritidis and 25 g/L for 20 minutes to S. Heidelberg. In the isolated Salmonella strains, lactic acid inhibited 97,89% of the strains and peracetic inhibited 100% of the strains. In contaminated chiller water, the compounds reduced the growth of standards strains. When applied to contaminated poultry meat, there was a reduction of Salmonella spp. 1,06 log10 CFU/g relative to the positive control with the use of sodium hypochlorite at 5,0 mg/L, 0,97 log10 CFU/g with dichloro and 0,56 log10 CFU/g with sodium hypochlorite 0,5 mg/L. For mesophiles reduction observed was 0,90 log10 CFU/g relative to the positive control with the use of sodium hypochlorite at 5,0 mg/L, 0,83 log10 CFU/g with dichloro and there isn´t reduction with hypochlorite with sodium 0,5 mg/L. For enterobacteriaceae reduction was 1,0 log10 CFU/g relative to the positive control with the use of sodium hypochlorite at 5,0 mg/L, 0,79 log10 CFU/g with dichloro and 0,22 log10 CFU/g with sodium hypochlorite at 0,5 mg/L. Lactic acid inhibit growth of the microorganisms tested. The data supports the discussions to regulate the use of the technology coadjuvants in the slaughter of poultry.

Optimization of lipase production from yeasts strains isolated from olive mil wastewater

Dissertação submetida à Universidade de Lisboa, Faculdade de Ciências para a obtenção do Grau de Mestre em Microbiologia Aplicada.

Avaliação da atividade antimicrobiana de filmes protéicos a base de anchoita (engraulis anchoita) incorporados com ácidos orgânicos.

Os filmes são produzidos a partir de macromoléculas, que podem ser utilizados como embalagem, como os polissacarídeos, lipídeos e proteínas. As proteínas se destacam dos demais, pois possuem uma estrutura com 20 monômeros diferentes, que confere um amplo potencial de ligações intermoleculares. A incorporação de agentes ativos em filmes é uma alternativa como embalagem, para inibir ou retardar a multiplicação de microrganismos patógenos e deteriorantes em alimentos. O objetivo deste trabalho foi avaliar a atividade antimicrobiana de filmes à base de isolado protéico de anchoita (Engraulis anchoita) – IPA adicionados de ácidos orgânicos. Para tanto, foi elaborado o IPA, pela solubilização alcalina da proteína e precipitação no ponto isoelétrico a partir de carne mecanicamente separada. O IPA foi avaliado quanto a sua composição proximal, aminoacídica e por DSC. A solução formadora dos filmes foi elaborada a partir de IPA, água, glicerol e hidróxido de sódio. As formulações dos filmes foram elaboradas segundo um planejamento fatorial 23 . Foram avaliadas as propriedades físico-químicas de resistência a tração (RT) e elongação (E); espessura, solubilidade e permeabilidade ao vapor de água (PVA); a diferença de cor (∆E*) e opacidade (Y) e microscopia eletrônica de varredura (MEV) de filmes à base de IPA. Os filmes com diferentes concentrações de ácido sórbico (AS) ou ácido benzóico (AB) foram desenvolvidos a partir da condição cujo as propriedades físico-químicas foram as melhores, sendo comparados aos filmes controles. Estes, foram avaliados quanto a sua atividade antimicrobiana frente aos microrganismos Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus e Salmonella Enteritidis pelo método de difusão em disco, além das propriedades físico-químicas, MEV e FT-IV. Os filmes com maior atividade antimicrobiana e os filmes controle foram aplicados sobre carne bovina, inoculados com os microrganismos inibidos no método de difusão em disco e armazenados a 5°C. Estes, foram avaliados a cada 2 dias durante 12 dias de armazenamento, pela método de contagem em gotas. O IPA apresentou 88,8% de proteína e 53,3% de aminoácidos polares e temperatura de desnaturação de 62,2°C. A espessura, PVA, ∆E* e Y dos filmes não foram afetados pelas variáveis estudadas no experimento. A menor solubilidade e maior RT dos filmes ocorreram em baixa concentração de IPA, glicerol e tratamento térmico, mas a E aumentou com o acréscimo dessas variáveis. As MEV das superfícies dos filmes foram homogêneas, para aqueles com leve tratamento térmico. O aumento da concentração de AS e AB na faixa de 0,50 a 1,50% resultou na diminuição da RT e aumento da E, solubilidade, ∆E* e Y. Houve mudança da organização molecular e interações intermoleculares entre as moléculas de IPA e AB testados pela avaliação do FT-IV. As MEV revelaram microporos em filmes com 1,50% de AS, o que resultou em filmes com menor homogeneidade. A maior atividade antimicrobiana foi verificada nos filmes com 1,50% de AS e AB frente a E. coli O157:H7, L. monocytogenes e S. Enteritidis. Estes filmes foram aplicados sobre carne bovina inoculada com E. coli O157:H7 e L. monocytogenes. Os filmes de AS frente a E. coli O157:H7 e L. monocytogenes apresentaram uma redução de 5 e 4 log UFC.g-1, respectivamente, em relação ao filme controle. O efeito do AB frente a estas bactérias, apresentou uma redução de 6 e 5 log UFC.g-1, ao final do 12° dia de armazenamento, respectivamente. Os filmes elaborados à base de IPA, adicionados de AS ou AB podem ser eficazes contra os patógenos alimentares testados.

Desenvolvimento e aplicação de modelo de calibração multivariada para determinação de açúcares e ácidos orgânicos em bebidas comerciais utilizando espectroscopia no infravermelho

The routine analysis for quantization of organic acids and sugars are generally slow methods that involve the use and preparation of several reagents, require trained professional, the availability of special equipment and is expensive. In this context, it has been increasing investment in research whose purpose is the development of substitutive methods to reference, which are faster, cheap and simple, and infrared spectroscopy have been highlighted in this regard. The present study developed multivariate calibration models for the simultaneous and quantitative determination of ascorbic acid, citric, malic and tartaric and sugars sucrose, glucose and fructose, and soluble solids in juices and fruit nectars and classification models for ACP. We used methods of spectroscopy in the near infrared (Near Infrared, NIR) in association with the method regression of partial least squares (PLS). Were used 42 samples between juices and fruit nectars commercially available in local shops. For the construction of the models were performed with reference analysis using high-performance liquid chromatography (HPLC) and refractometry for the analysis of soluble solids. Subsequently, the acquisition of the spectra was done in triplicate, in the spectral range 12500 to 4000 cm-1. The best models were applied to the quantification of analytes in study on natural juices and juice samples produced in the Paraná Southwest Region. The juices used in the application of the models also underwent physical and chemical analysis. Validation of chromatographic methodology has shown satisfactory results, since the external calibration curve obtained R-square value (R2) above 0.98 and coefficient of variation (%CV) for intermediate precision and repeatability below 8.83%. Through the Principal Component Analysis (PCA) was possible to separate samples of juices into two major groups, grape and apple and tangerine and orange, while for nectars groups separated guava and grape, and pineapple and apple. Different validation methods, and pre-processes that were used separately and in combination, were obtained with multivariate calibration models with average forecast square error (RMSEP) and cross validation (RMSECV) errors below 1.33 and 1.53 g.100 mL-1, respectively and R2 above 0.771, except for malic acid. The physicochemical analysis enabled the characterization of drinks, including the pH working range (variation of 2.83 to 5.79) and acidity within the parameters Regulation for each flavor. Regression models have demonstrated the possibility of determining both ascorbic acids, citric, malic and tartaric with successfully, besides sucrose, glucose and fructose by means of only a spectrum, suggesting that the models are economically viable for quality control and product standardization in the fruit juice and nectars processing industry.

Crassaminicella profunda gen. nov., sp nov., an anaerobic marine bacterium isolated from deep-sea sediments

A novel, anaerobic, chemo-organotrophic bacterium, designated strain Ra1766HT, was isolated from sediments of the Guaymas basin (Gulf of California, Mexico) taken from a depth of 2002 m. Cells were thin, motile, Gram-stain-positive, flexible rods forming terminal endospores. Strain Ra1766H(T) grew at temperatures of 25-45 degrees C (optimum 30 degrees C), pH 6.7-8.1 (optimum 7.5) and in a salinity of 5-60 g l(-1) NaCl (optimum 30 g l(-1)). It was an obligate heterotrophic bacterium fermenting carbohydrates (glucose and mannose) and organic acids (pyruvate and succinate). Casamino acids and amino acids (glutamate, aspartate and glycine) were also fermented. The main end products from glucose fermentation were acetate, butyrate, ethanol, H-2 and CO2. Sulfate, sulfite, thiosulfate, elemental sulfur, fumarate, nitrate, nitrite and Fe(III) were not used as terminal electron acceptors. The predominant cellular fatty acids were C-14 : 0, C-16:1 omega 7, C-16:1 omega 7 DMA and C-16:0. The main polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and phospholipids. The G +C content of the genomic DNA was 33.7 molo/o. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain Ra1766H(T) was affiliated to cluster XI of the order Clostridia les, phylum Firmicutes. The closest phylogenetic relative of Ra1766H(T) was Geosporobacter subterraneus (94.2% 16S rRNA gene sequence similarity). On the basis of phylogenetic inference and phenotypic properties, strain Ra1766H(T) (=DSM 27501(T)=JCM 19377(T)) is proposed to be the type strain of a novel species of a novel genus, named Crassaminicella pro funda.

Alteration of cell-wall porosity is involved in osmotic stress-induced enhancement of aluminium resistance in common bean (Phaseolus vulgaris L.)

Aluminium (Al) toxicity and drought are the two major abiotic stress factors limiting common bean production in the tropics. Using hydroponics, the short-term effects of combined Al toxicity and drought stress on root growth and Al uptake into the root apex were investigated. In the presence of Al stress, PEG 6000 (polyethylene glycol)-induced osmotic (drought) stress led to the amelioration of Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1. PEG 6000 (>> PEG 1000) treatment greatly decreased Al accumulation in the 1 cm root apices even when the roots were physically separated from the PEG solution using dialysis membrane tubes. Upon removal of PEG from the treatment solution, the root tips recovered from osmotic stress and the Al accumulation capacity was quickly restored. The PEG-induced reduction of Al accumulation was not due to a lower phytotoxic Al concentration in the treatment solution, reduced negativity of the root apoplast, or to enhanced citrate exudation. Also cell-wall (CW) material isolated from PEG-treated roots showed a low Al-binding capacity which, however, was restored after destroying the physical structure of the CW. The comparison of the Al(3+), La(3+), Sr(2+), and Rb(+) binding capacity of the intact root tips and the isolated CW revealed the specificity of the PEG 6000 effect for Al. This could be due to the higher hydrated ionic radius of Al(3+) compared with other cations (Al(3+) >> La(3+) > Sr(2+) > Rb(+)). In conclusion, the results provide circumstantial evidence that the osmotic stress-inhibited Al accumulation in root apices and thus reduced Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1 is related to the alteration of CW porosity resulting from PEG 6000-induced dehydration of the root apoplast.

Madeira wine vinification process: analysis and control

The present study was done in collaboration with J. Faria e Filhos company, a Madeira wine producer, and its main goal was to fully characterize three wines produced during 2014 harvest and identify possible improving points in the winemaking process. The winemaking process was followed during 4 weeks, being registered the amounts of grapes received, the fermentation temperatures, the time at which fermentation was stopped and evolution of must densities until the fortification time. The characterization of musts and wines was done in terms of density, total and volatile acidity, alcohol content, pH, total of polyphenol, organic acids composition, sugars concentration and the volatile profile. Also, it was developed and validated an analytical methodology to quantify the volatile fatty acids, namely using SPME-GC-MS. Briefly, the following key features were obtained for the latter methodology: linearity (R2=0.999) e high sensitivity (LOD =0.026-0.068 mg/L), suitable precision (repeatability and reproducibility lower than 8,5%) and good recoveries (103,11-119,46%). The results reveal that fermentation temperatures should be controlled in a more strictly manner, in order to ensure a better balance in proportion of some volatile compounds, namely the esters and higher alcohols and to minimize the concentration of some volatiles, namely hexanoic, octanoic and decanoic acids, that when above their odours threshold are not positive for the wine aroma. Also, regarding the moment to stop the fermentation, it was verified that it can be introduced changes which can also be benefit to guarantee the tipicity of Madeira wine bouquet.

Control of grey mould and stem-end rot in strawberry plants growing in a subtropical environment

The effect of different fungicide programs on grey mould (caused by Botrytis cinerea) and stem-end rot (caused by Gnomoniopsis fructicola) affecting strawberry plants (Fragaria ×ananassa cv. Festival) was studied in subtropical Australia over three years. The treatments involved a range of different synthetic multi- and single-site fungicides with different modes of action, a plant-defence promoter, plant extracts (lupin and rhubarb), organic acids, fatty acids, a salt, two strains of Bacillus subtilis, and single strains of B. amyloliquefaciens, Streptomyces lydicus and Trichoderma harzianum. Standard programs based on captan and thiram alternated, and applied with iprodione, fenhexamid, cyprodinil + fludioxonil, and penthiopyrad resulted in 3–4 % of unmarketable fruit compared with 25–38 % in the water-treated controls. There was no difference in the level of disease suppression when five or thirteen applications of single-site fungicides were rotated with the two multi-site fungicides. The incidence of unmarketable fruit was similar to the standard programs using isopyrazam (in 1 year out of 2), or penthiopyrad, fluazinam, chlorothalonil or thiram alone (in 1 year out of 1). The other fungicide programs were generally less effective. There were strong relationships between marketable yield and the incidence of unmarketable fruit over the three years (R2s = 0.82–0.93). A strategy based on thiram and captan applied alternately, with reduced applications of single-site fungicides is recommended and should reduce the chance of resistance to single-site fungicides becoming widespread in populations of the grey mould fungus. Although the program based on thiram alone had a similar incidence of unmarketable fruit as the standard program, repeated weekly applications of thiram are not recommended as they may cause unacceptable residues in the fruit. There were issues with some of the other fungicides due to phytotoxicity, residues, or difficulties with registering new fungicides that are in the same chemical group as currently registered products.

Composição química e atividade antioxidante de maracujá-roxo (Passiflora edulis Sims edulis)em diferentes graus de maturação

O maracujá-roxo é um fruto tropical que tem ganho destaque devido ao seu valor nutricional, organolético e às emergentes descobertas acerca das suas propriedades farmacêuticas e antioxidantes. Durante o processo de maturação decorrem várias reações de ordem física e química, e no intuito de enriquecer o conhecimento acerca de como a composição química e as propriedades do maracujá-roxo evoluem ao longo da maturação, este trabalho consistiu na caracterização física do fruto inteiro e na caracterização química geral das sementes, casca e polpa em frutos separados em cinco graus de maturação diferentes (G1 a G5). Foi também avaliada a atividade antioxidante (atividade antiradicalar e poder redutor), e o teor em compostos fenólicos (fenóis totais, derivados do ácido hidroxicinâmico e flavonóis) das diferentes matrizes que compõem o fruto ao longo da maturação. A partir dos resultados obtidos verificou-se na casca que o teor em cinza bruta aumenta essencialmente entre G1 e G2 enquanto se verificou uma diminuição da proteína. Em relação à semente, o teor em cinza bruta aumenta gradualmente, e o teor em proteína aumenta de G1 para G2, estabilizando posteriormente nos 8% (base seca). O teor em gordura aumenta gradualmente ao longo da maturação, verificando-se um maior acumulo entre G1 e G2 (9,9 e 19,1% respetivamente). No que respeita à polpa, há uma diminuição nos teores de cinza e proteína e aumento dos sólidos solúveis totais.Verificou-se um aumento ligeiro do pH ao longo da maturação (entre 2,8 a 3,1) e uma diminuição da acidez (entre 12,1 e 6,7 g de ácido cítrico 100 mL-1). Constatou-se que os açúcares predominantes na polpa foram a sacarose, frutose e glucose. Quanto aos ácidos orgánicos, o ácido cítrico foi o maioritário em todos os graus de maturação e teores mais baixos foram quantificados para os ácidos málico e ascórbico. Entre as diferentes partes do fruto estudadas, as cascas foram a matriz mais antioxidante, aumentando o seu potencial bioativo durante a maturação. As sementes apresentaram valores mais elevados de fenóis totais, derivados do ácido hidroxicinâmico e flavonóis. Foi verificado que a atividade antioxidante esteve correlacionada com os valores de fenóis totais presentes nas diferentes partes do fruto ao longo da maturação. De acordo com o conhecimento dos autores, este é o primeiro estudo que toma em consideração as alterações sofridas pelas diferentes partes do maracujá-roxo produzido em Portugal, ao longo da maturação.

Dietary supplements based on borututu: wich is the most suitable choice for bioactive purposes?

Borututu ( Cochlospermum angolensis Welw.) is a widespread tree in Angola used since antiquity by traditional healers for the prevention and treatment of hepatic diseases and for the prophylaxis of malaria [1]. This plant is mostly consumed as infusions but is also available as dietary supplements, such as piiis, capsules, and syrups, among others. In the present study, the aim was to evaluate the proximate composition and energetic contribution of borututu as weii as its composition in hydrophilic (sugars and organic acids) and lipophilic (fatty acids and tocopherols) compounds, given the fact that this plant is directly introduced in some dietary supplements. Furthermore, the bioactivity (antioxidant, hepatoprotective and antimicrobial activities) of three different formulations of borututu (infusion, pills, and syrup) was assessed and compared, and since plant beneficial properties are often ascribed to phenolic compounds [2], the phenolic profile of the formulations was also analysed. Carbohydrates (88 g/100 g) and fat (2.5 g/100 g) were the major and tl1e minor components of the studied borututu dry barks, respectively, with an energetic contribution of 384 kcal/100 g. Fructose was the most abundant sugar (1.3 g/100 g), foilowed by sucrose, trehalose and glucose (1.1, 0.98 and 0.79 g/100 g, respectively). Oxalic (0.70 g/100 g), malic (0.63 g/100 g) and citric (0.57 g/100 g) acids were present in higher amounts but shikimic and fumaric acids were also detected. Among the fatty acids found in borututu, a prevalence of saturated fatty acids (SF A; 48.2%) was observed, whereas polyunsaturated (PUFA) and monounsaturated (MUFA) fatty acids were detected in relative percentages of 30.9% and 20.8%, respectively. P-tocopherol was the most abundant of the four isoforms found in the sample, foiiowed by o-, a- and y-tocopherol, present in concentrations of 597,43, 3.7 and 2.0 g/100 g, respectively. Borututu infusion revealed the highest antioxidant activity, with EC50 values ranging from 20 to 600 J.lg/mL and was the only formulation inhibiting the growth of an HepG2 ceii line, with a Gl5o value of 146 J.lg/mL. This formulation.also revealed the best antimicrobial capacity and proved to be able to inhibit the growth of Escherichia coli, E. coli ESBL, Staphylococcus aureus and Pseudomonas aeruginosa, with MIC values of 50, 6.2, 1.6 and 25 mg!mL, respectively. Pills revealed activity against some of the studied bacterial strains and the syrup did not reveal antimicrobial activity at the studied concentration. Eilagic acids, methyl ellagic acids, eucaglobulinlglobulusin B and (epi)gaiiocatechin-0-gallate were the compounds present in all the different formulations. The highest concentration of phenolic compounds was found in the infusion extract. Protocatechuic acid was the most abundant phenolic compound in the infusions, the only preparation where it was detected, whereas ( epi)gaiiocatechin- 0-gallate was the main phenolic in the pills and eucaglobulinlglobulusin in the syrup. In a general way, borututu proved to be a good source of phytochemicals such as phenolic compounds, with the infusions revealing the best bioactive properties.

From the field to the table: ionizing radiation as a feasible postharvest treatment for fresh and dried plant foods

Food irradiation is a treatment that involves subjecting in-bulk or packaged food to a controlled dose of ionizing radiation, with a clearly defined goal. It has been used for disinfestation and sanitization of food commodities and to retard postharvest ripening and senescence processes, being a sustainable alternative to chemical agents 1 . Doses up to 10 kGy are approved by several international authorities for not offering negative effects to food from a nutrition and toxicology point of view 2 . However, the adoption of this technology for food applications has been a slow process due to some misunderstandings by the consumer who often chooses non-irradiated foods. In this study, the effects of the ionizing radiation treatment on physical, chemical and bioactive properties of dried herbs and its suitability for preserving quality attributes of fresh vegetables during cold storage were evaluated. The studied herbs, perennial spotted rockrose (Tuberaria lignosa (Sweet) Samp.) and common mallow (Malva neglecta Wallr.) were freeze-dried and then irradiated up to 10 kGy in a Cobalt-60 chamber. The selected vegetables, watercress (Nasturtium officinale R. Br.) and buckler sorrel (Rumex induratus Boiss. Reut.) were rinsed in tap water, packaged in polyethylene bags, submitted to irradiation doses up to 6 kGy and then were stored at 4 C for a period of up to 12 days. Physical, chemical and bioactive parameters of irradiated and non-irradiated samples were evaluated using different methodologies the colour was measured with a colorimeter, individual chemical compounds were analyzed by chromatographic techniques, antioxidant properties were evaluated using in vitro assays based on different reaction mechanisms, and other quality analyses were performed following official methods of analysis. The irradiation treatment did not significantly affect the colour of the perennial spotted rockrose samples, or its phenolic composition and antioxidant activity 3 . Medium doses preserved the colour of common mallow and a low dose did not induce any adverse effect in the organic acids profile. The green colour of the irradiated vegetables was maintained during cold storage but the treatment had pros and cons in other quality attributes. The 2 kGy dose preserved free sugars and favoured polyunsaturated fatty acids (PUFA) while the 5 kGy dose favoured tocopherols and preserved the antioxidant properties in watercress samples. The 6 kGy dose was a suitable option for preserving PUFA and the ω-6 ω-3 fatty acids ratio in buckler sorrel samples. This comprehensive experimental work allowed selecting appropriate processing doses for the studied plant foods in order to preserve its quality attributes and edibility.