Improvement of adhesion and paint ability of EVA copolymers with different vinyl acetate contents by treatment with UV-ozone

The surface modifications produced by UV-ozone treatment of two ethylene-vinyl acetate (EVA) copolymers containing 12 and 20 wt% vinyl acetate (EVA12 and EVA20 respectively) were studied. The treatment with UV-ozone improved the wettability of both EVAs due to the creation of new carbon–oxygen moieties. The extent of these modifications increased with increasing length of the treatment and the modifications produced in EVA20 were produced for shorter lengths of treatment. The UV-ozone treatment also created roughness and heterogeneities on the EVA surfaces. Whereas roughness formation prevailed on the UV-ozone treated EVA12, important ablation was dominant on the treated EVA20. T-peel strength values in joints made with polychloroprene adhesive increased when the EVAs were treated with UV-ozone. Short length of UV-ozone treatment (1 min) produced higher T-peel strength in joints made with EVA20 whereas higher T-peel strength values in joints made with EVA12 were obtained after treatment for 5–7.5 min in which a cohesive failure into a weak boundary layer on the treated EVA surface was found. Furthermore, the adhesion of UV-ozone treated EVA20 to acrylic paint increased. Finally, the ageing resistance of the treated EVA/polychloroprene adhesive joints was good and the surface modifications on the UV-ozone treated EVAs lasted for 24 h after treatment at least.Financial support from Research Spanish Agency MICYT (projects MAT2002-02463 and PETRI95-0578-OP) is acknowledged

Estudio y caracterización molecular de la producción de aminas biógenas por parte de bacterias lácticas de origen enológico.

RESUMEN Dada la influencia negativa de la presencia de aminas biógenas en el vino, por el efecto perjudicial sobre la salud humana y por la disminución de la calidad del vino y dada también la presencia de las bacterias lácticas (BL) en los mismos, en esta tesis nos planteamos determinar la importancia de las BL en la presencia de estas aminas en el vino, así como que factores podían influir en la concentración de las mismas o en la mayor o menor producción por parte de las BL. En primer lugar, desarrollamos un método enzimático que permite analizar la concentración de histamina en vino y la producción de la misma por BL, a continuación analizamos la concentración de histamina, tiramina, feniletilamina, putrescina, cadaverina y triptamina en vinos de distintas zonas geográficas y de distintas variedades de uva, así como la influencia del pH, de la fermentación maloláctica y de los periodos de almacenamiento, además, la capacidad aminobiogénica de bacterias aisladas de algunos vinos también fue analizada. Entre otros resultados, pudimos comprobar que era durante la fermentación maloláctica cuando incrementaban los niveles de histamina, tiramina y feniletilamina, pero no de las otras aminas estudiadas, además encontramos BL capaces de producir histamina, tiramina y feniletilamina, pero no putrescina, cadaverina ni triptamina. Ante la necesidad de desarrollar un método molecular que nos permitiera identificar la presencia del gen de la tirosina descarboxilasa (tdc) en BL de origen enológico, nos planteamos la secuenciación y caracterización molecular del operón tdc del Lactobacillus brevis 9809, encontrándonos que este operón contiene 4 genes completos que codifican para una tirosin-RNAt sintetasa, para la tirosina descarboxilasa, para una probable tirosina permeasa y para un antiporter Na+/H+. Posteriormente, analizamos mediante métodos moleculares y bioquímicos la capacidad de producción de histamina, tiramina y feniletilamina por parte de más de 150 cepas de BL, además de correlacionar la presencia en vinos de estas aminas con la capacidad de producción de las mismas por las BL presentes en los mismos, observamos que Lactobacillus hilgardii y Pediococcus parvulus eran los responsables de los elevados niveles de histamina en vino y que la mayor parte de los Oenococcus podían producir esta amina, pero no representaban un peligro al producir bajos niveles. En cuanto a la tiramina, todos los Lactobacillus brevis y algún Lactobacillus hilgardii producían tiramina y feniletilamina atribuyéndose a estas especies la responsabilidad de los niveles de tiramina y feniletilamina. Finalmente, estudiamos la influencia de factores físico-químicos del vino y la fase de crecimiento sobre la producción de histamina, pudimos comprobar como la glucosa y fructosa y los ácidos málico y cítrico junto con la histamina disminuían la expresión del gen de la histidina descarboxilasa y por tanto la producción de histamina, por otro lado la histidina aumentaba dicha expresión, además la presencia de ciertos niveles de etanol y pyridoxal 5-fosfato aumentaba la producción de histamina actuando sobre el enzima histidina descarboxilasa, por último observamos como la fase de crecimiento de las BL influye decisivamente en la expresión del gen de la histidina descarboxilasa. __________________________________________________________________________________________________Biogenic amines are organic bases endowed with biological activity which are frequently found in fermented foods and beverages. They are produced mainly as a consequence of decarboxylation of amino acids. High concentrations of biogenic amines can cause undesirable physiological effects in sensitive humans, especially when alcohol and acetaldehyde are present In this work, we describe a new enzymatic method that allows directly estimating histamine concentration in synthetic media, grape must or wine (white, rosé and red) without polyphenols or sugar interferences. This new enzymatic method shows a good correlation (r=0.996, p<0.001) between the histamine concentrations and absorbances in the interval 0.4 to 160 mg l-1. The influence of grape variety, type of vinification, wine pH, malolactic fermentation, and storage in bottle on biogenic amine concentrations was also studied. Results show important differences in putrescine and histamine concentrations between regions, varieties of grape and type of wine; differences were less appreciable for the remaining biogenic amines studied. Low pH prevented biogenic amine formation. Malolactic fermentation and short storage periods in bottle (3 - 6 months) showed increases in histamine concentration, while longer periods of storage led to a general decrease in histamine. Grape varieties, different types of winemaking, pH and lactic acid bacteria may be responsible for the differences observed in the biogenic amine concentration of wines analyzed. Strains of lactic acid bacteria were isolated in this work, and their ability to form biogenic amines was assayed in synthetic media, grape must and wine. The species exhibiting the highest frequency of histamine production is Oenococcus oeni. However, the concentration of histamine produced by this species is lower than the produced by strains belonging to species of Lactobacillus and Pediococcus. A correlation of 100% between presence of histidine decarboxylase gene and histamine production was observed. The main wine species tyramine and phenylethylamine producer was Lactobacillus brevis whereas this character was no present or infrequent in the rest of analysed wine strains. The sequence of tyrosine decarboxylase operon contained four complete genes encoding atyrosyl-tRNA synthetase, the tyrosine decarboxylase, a probable tyrosine permease and a Na+/H+ antiporter. The aim of this work was also to study the influence of enological factors (organic acids, sugars, ethanol, histidine, histamine, SO2, pH and temperature) on the hdc expression and on HDC activity in Lactobacillus hilgardii, Pediococcus parvulus and Oenococcus oeni. Cell extracts and whole cells were used to study the influence of different factors on histidine decarboxylase gene (hdc) expression and histidine decarboxylase enzyme (HDC) activity. Glucose, fructose, malic acid, citric acid and histamine diminished the hdc expression, while ethanol and histidine increased the histidine decarboxylase enzyme activity. Temperature and pH had effect on the activity of HDC but not on hdc expression. Tartaric acid and L-lactic acid, and SO2 had no effect on enzyme synthesis and activity. Lb hilgardii is the most active, and O. oeni the less one, lactic acid bacteria species difer in the relative enzymatic activity but all the factors affected in the same way to Lb. hilgardii, P. parvulus and O. oeni. The hdc gene expression was lowered by glucose, fructose, malic acid, and citric acid, whereas ethanol enhanced the HDC enzyme activity. This behavior was observed for all the studied microorganisms. Thus, the conditions that normally occur during malolactic fermentation and later on, could favor the histamine production. SO2 could prevent bacterial growth, but does not diminish the HDC enzyme activity

Phytoestrogen Metabolism by Adult Human Gut Microbiota

Phytoestrogens are plant-derived polyphenols with a structure similar to human estrogens. The three main groups of phytoestrogens, isoflavones, ellagitannins, and lignans, are transformed into equol, urolithins, and enterolignans, respectively, by bacteria. These metabolites have more estrogenic/antiestrogenic and antioxidant activities than their precursors, and they are more bioavailable. The aim of this study was to analyze the metabolism of isoflavones, lignans and ellagitannins by gut microbiota, and to study the possible correlation in the metabolism of these three groups of phytoestrogens. In vitro fermentation experiments were performed with feces samples from 14 healthy adult volunteers, and metabolite formation was measured by HPLC-PAD and HPLC-ESI/MS. Only the microbiota of one subject produced equol, while most of them showed production of O-desmethylangolensin (O-DMA). Significant inter-subject differences were observed in the metabolism of dihydrodaidzein and dihydrogenistein, while the glucoside isoflavones and their aglycones showed less variability, except for glycitin. Most subjects produced urolithins M-5 and E. Urolithin D was not detected, while uroltithin B was found in half of the individuals analyzed, and urolithins A and C were detected in two and four subjects, respectively. Enterolactone was found in all subjects, while enterodiol only appeared in five. Isoflavone metabolism could be correlated with the metabolism of lignans and ellagitannins. However, the metabolism of ellagitannins and lignans could not be correlated. This the first study where the metabolism of the three groups together of phytoestrogen, isoflavones, lignans, and ellagitannins by gut microbiota is analyzed

Characterization of a benzyl alcohol dehydrogenase from Lactobacillus plantarum WCFS1

Aroma is an important sensory parameter of food products. Lactic acid bacteria have enzymatic activities that could be important in the modification of food aroma. The complete genome sequence from Lactobacillus plantarum WCFS1 shows a gene (lp_3054) putatively encoding a protein with benzyl alcohol dehydrogenase activity. To confirm its enzymatic activity lp_3054 from this strain has been overexpressed and purified. Protein alignment indicated that lp_3054 is a member of the family of NAD(P)-dependent long-chain zinc-dependent alcohol dehydrogenases. In lp_3054 all of the residues involved in zinc and cofactor binding are conserved. It is also conserved the residue that determines the specificity of the dehydrogenase toward NAD+ rather than NADP+ and, therefore, L. plantarum benzyl alcohol dehydrogenase is less active in the presence of NADP+ than in the presence of NAD+. The purified enzyme exhibits optimal activity at pH 5.0 and 30 °C. The kinetic parameters Km and Vmax on benzyl alcohol as a substrate were, respectively, 0.23 mM and 204 μmol h-1 mg-1. Besides its activity toward benzyl alcohol, it showed activity against nerol, geraniol, phenethyl alcohol, cinnamyl alcohol, and coniferyl alcohol, all of which are volatile compounds involved in determining food aroma. The biochemical demonstration of a functional benzyl alcohol dehydrogenase activity in this lactic acid bacteria species should be considered when the influence of bacterial metabolism in the aroma of food products is determinedL. plantarum WCFS1 strain was kindly provided by M. Kleerebezem from the Wageningen Centre for Food Sciences, NIZO Food Research. The technical assistance of M. V. Santamaría is greatly appreciatedPeer reviewe

Degradación de compuestos fenólicos presentes en vino mediante cepas de Lactobacillus plantarum

Trabajo presentado al IX Congreso Nacional de Investigación Enológica (GIENOL) celebrado en Badajoz del 29 de mayo al 1 de junio de 2007.Los compuestos fenólicos influyen en las características sensoriales y nutricionales del vino. La especie Lactobacillus plantarum, modelo de cultivo iniciador en biotecnología de alimentos vegetales, es una de las especies de bacterias lácticas mayoritarias en vino. El estudio pretende analizar la capacidad que presentan varias cepas de L. plantarum para degradar 15 de los compuestos fenólicos mayoritarios presentes en vinos.Este trabajo ha sido realizado gracias a la financiación recibida de los proyectos AGL2005-00470 (CICYT), RM03-002 (INIA) y 07G/0035/2003 (CAM)Peer reviewe

Study of the inhibitory activity of phenolic compounds found in olive products and their degradation by Lactobacillus plantarum strains

Lactobacillus plantarum is the main species responsible for the spontaneous fermentation of Spanish-style green olives. Olives and virgin oil provide a rich source of phenolic compounds. This study was designed to evaluate inhibitory growth activities of nine olive phenolic compounds against four L. plantarum strains isolated from different sources, and to explore the L. plantarum metabolic activities against these phenolic compounds. None of the nine compounds assayed (oleuropein, hydroxytyrosol, tyrosol, as well as vanillic, p-hydroxybenzoic, sinapic, syringic, protocatechuic and cinnamic acids) inhibited L. plantarum growth at the concentration found in olive products. Oleuropein and tyrosol concentrations higher than 100 mM were needed to inhibit L. plantarum growth. On the other hand, sinapic and syringic acid showed the highest inhibitory activity since concentrations ranging from 12.5 to 50 mM inhibited L. plantarum growth in all the strains analyzed. Among the nine compounds assayed, only oleuropein and protocatechuic acid were metabolized by L. plantarum strains grown in the presence of these compounds. Oleuropein was metabolized mainly to hydroxytyrosol, while protocatechuic acid was decarboxylated to catechol. Metabolism of oleuropein was carried out by inducible enzymes since a cell-free extract from a culture grown in the absence of oleuropein was unable to metabolize it. Independent of their isolation source, the four L. plantarum strains analysed showed similar behaviour in relation to the inhibitory activity of phenolic compounds, as well as their ability to metabolize these compoundsThis work was supported by grants AGL2005-00470 (CICYT), FUN-C-FOOD Consolider 25506 (MEC), RM03-002 (INIA), and S-0505/AGR/000153 (CAM). J.M. Landete and J. A. Curiel are recipients of a postdoctoral and predoctoral fellowship, respectively, from the MEC. H. Rodríguez is a recipient of a predoctoral fellowship from the I3P-CSIC.Peer reviewe

Alcohol Consumption during Pregnancy : Analysis of Two Direct Metabolites of Ethanol in Meconium

Alcohol consumption in young women is a widespread habit that may continue during pregnancy and induce alterations in the fetus. We aimed to characterize prevalence of alcohol consumption in parturient women and to assess fetal ethanol exposure in their newborns by analyzing two direct metabolites of ethanol in meconium. This is a cross-sectional study performed in September 2011 and March 2012 in a series of women admitted to an obstetric unit following childbirth. During admission, socio-demographic and substance use (alcohol, tobacco, cannabis, cocaine, and opiates) during pregnancy were assessed using a structured questionnaire and clinical charts. We also recorded the characteristics of pregnancy, childbirth, and neonates. The meconium analysis was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to detect the presence of ethyl glucuronide (EtG) and ethyl sulfate (EtS). Fifty-one parturient and 52 neonates were included and 48 meconium samples were suitable for EtG and EtS detection. The median age of women was 30 years (interquartile range (IQR): 26-34 years); EtG was present in all meconium samples and median concentration of EtG was 67.9 ng/g (IQR: 36.0-110.6 ng/g). With respect to EtS, it was undetectable (<0.01 ng/g) in the majority of samples (79.1%). Only three (6%) women reported alcohol consumption during pregnancy in face-to-face interviews. However, prevalence of fetal exposure to alcohol through the detection of EtG and EtS was 4.2% and 16.7%, respectively. Prevention of alcohol consumption during pregnancy and the detection of substance use with markers of fetal exposure are essential components of maternal and child health

Lactation curves in captive Iberian red deer (Cervus elaphus hispanicus)

This study examines milk production and the effect of milk production and sex of calf on body weights and gains of red deer calves and hinds of the Iberian subspecies (Cervus elaphus hispanicus). Milk production was assessed in 14 hinds by weighing calves before and after suckling and by adjusting these values to the Gamma function. Gamma estimates of total milk production up to d 105 were similar to the amounts computed directly from double weighing. Hinds showed two types of lactation curve: 1) the standard mammal lactation curve, with an asymmetrical peak at wk 2 to 4 (Type I) and 2) decreasing curves with no peak (Type II). Although there was great interindividual variability, hinds with Type I curves showed a trend to produce more milk than those with Type II. The type of curve did not seem to affect weight variables of the calf or those of the dam. Calves that gained more weight consumed greater amounts of milk (P < .05). Males were heavier than females at birth (P < .05), but males did not differ from females in their weight at 105 d, milk consumption, and gain. Gender did not affect hind weight, but dams of male calves showed a trend to be lighter (P = .063) at d 105 than dams of female calves. Our results suggest that suckling differences found in other studies between male and female calves may not involve differences in milk production, although other rearing costs seem to affect hind weight losses. They also suggest that the curve type may not depend only, as reported, on the nutrition plane

Effect of fermented soy beverage in aged female mice model

11 Pág.Soy beverage is a rich source of phytoestrogens isoflavones, with potential benefits on health. The effect of those compounds depends greatly on their bacterial metabolization into their aglycone forms. This study evaluated the health effects of two soy beverages, non-fermented (SB) and fermented with Bifidobacterium pseudocatenulatum INIA P815 (FSB), in acyclic and cyclic C57BL/6J aged female mice as a model of menopause and premenopause, respectively. SB and FSB treatments were administrated for 36 days and, subsequently, body weight, lipid and inflammatory profile and fertility were analyzed and compared. In addition, hepatic gene expression and faecal microbiota composition were also assessed. After fermentation, FSB presented a high content in the aglycones daidzein and genistein and a higher antioxidant activity. FSB treated cyclic mice showed a significant increase in the number of retrieved oocytes and zigotes. Differences in serum lipids were observed in triglycerides, which were lower in FSB than in SB groups. None of the treatments influenced the inflammatory profile or caused a dramatic change in the intestinal microbiota profile or hepatic gene expression in any of the groups. Our data showed that FSB provided greater health benefits than SB in lipid profile and fertility in cyclic mice. These beneficial effects could be attributed to the fermentation process, which produces more bioavailable and bioactive compounds, achieving a greater impact on health.This work was supported by projects RTA2017-00002-00-00, PID2020-11960RB-I00, and RTI2018-093548-B-I00 from the Spanish Ministry of Science and Innovation. We appreciate the help of Doctors Javier Ortego and Eva Calvo from CISA-CSIC in the analysis of inflammatory profile. We are grateful to the ICTAN (Institute of Food Science, Technology and Nutrition, Madrid, Spain) Analysis Services Unit for providing chromatography and mass spectrometry facilities.Peer reviewe

Probiotic and Functional Properties of Limosilactobacillus reuteri INIA P572

Limosilactobacillus reuteri INIA P572 is a strain able to produce the antimicrobial compound reuterin in dairy products, exhibiting a protective effect against some food-borne pathogens. In this study, we investigated some probiotic properties of this strain such as resistance to gastrointestinal passage or to colonic conditions, reuterin production in a colonic environment, and immunomodulatory activity, using different in vitro and in vivo models. The results showed a high resistance of this strain to gastrointestinal conditions, as well as capacity to grow and produce reuterin in a human colonic model. Although the in vitro assays using the RAW 264.7 macrophage cell line did not demonstrate direct immunomodulatory properties, the in vivo assays using a Dextran Sulphate Sodium (DSS)-induced colitic mice model showed clear immunomodulatory and protective effects of this strain.This work was supported by project no. RTA2017-00002-00-00 from the Spanish Ministry of Science and Innovation, by the Junta de Andalucía (CTS 164) and Instituto de Salud Carlos III (PI19/01058) with funds from the European Union.Ye