Bio-detoxification of mycotoxins by lactic acid bacteria from different food matrices

Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP- PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 g/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 m pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Os mass media como agentes educadores para a saúde

Dissertação de mestrado em Ciências da Comunicação (área de especialização em Informação e Jornalismo)

Aromatic amines sources, environmental impact and remediation

Aromatic amines are widely used industrial chemicals as their major sources in the environment include several chemical industry sectors such as oil refining, synthetic polymers, dyes, adhesives, rubbers, perfume, pharmaceuticals, pesticides and explosives. They result also from diesel exhaust, combustion of wood chips and rubber and tobacco smoke. Some types of aromatic amines are generated during cooking, special grilled meat and fish, as well. The intensive use and production of these compounds explains its occurrence in the environment such as in air, water and soil, thereby creating a potential for human exposure. Since aromatic amines are potential carcinogenic and toxic agents, they constitute an important class of environmental pollutants of enormous concern, which efficient removal is a crucial task for researchers, so several methods have been investigated and applied. In this chapter the types and general properties of aromatic amine compounds are reviewed. As aromatic amines are continuously entering the environment from various sources and have been designated as high priority pollutants, their presence in the environment must be monitored at concentration levels lower than 30 mg L1, compatible with the limits allowed by the regulations. Consequently, most relevant analytical methods to detect the aromatic amines composition in environmental matrices, and for monitoring their degradation, are essential and will be presented. Those include Spectroscopy, namely UV/visible and Fourier Transform Infrared Spectroscopy (FTIR); Chromatography, in particular Thin Layer (TLC), High Performance Liquid (HPLC) and Gas chromatography (GC); Capillary electrophoresis (CE); Mass spectrometry (MS) and combination of different methods including GC-MS, HPLC-MS and CE-MS. Choosing the best methods depend on their availability, costs, detection limit and sample concentration, which sometimes need to be concentrate or pretreated. However, combined methods may give more complete results based on the complementary information. The environmental impact, toxicity and carcinogenicity of many aromatic amines have been reported and are emphasized in this chapter too. Lately, the conventional aromatic amines degradation and the alternative biodegradation processes are highlighted. Parameters affecting biodegradation, role of different electron acceptors in aerobic and anaerobic biodegradation and kinetics are discussed. Conventional processes including extraction, adsorption onto activated carbon, chemical oxidation, advanced oxidation, electrochemical techniques and irradiation suffer from drawbacks including high costs, formation of hazardous by-products and low efficiency. Biological processes, taking advantage of the naturally processes occurring in environment, have been developed and tested, proved as an economic, energy efficient and environmentally feasible alternative. Aerobic biodegradation is one of the most promising techniques for aromatic amines remediation, but has the drawback of aromatic amines autooxidation once they are exposed to oxygen, instead of their degradation. Higher costs, especially due to power consumption for aeration, can also limit its application. Anaerobic degradation technology is the novel path for treatment of a wide variety of aromatic amines, including industrial wastewater, and will be discussed. However, some are difficult to degrade under anaerobic conditions and, thus, other electron acceptors such as nitrate, iron, sulphate, manganese and carbonate have, alternatively, been tested.

Molecular mass distribution of materials solubilized by xylanase treatment of Douglas-Fir kraft pulp

Irgazyme, a commercial xylanase preparation from Trichoderma longibrachiatum, and xylanase D a purified enzyme from Trichoderma harzianum E58 were tested for their ability to enhance peroxide bleaching of Douglas-fir (Pseudotsuga menziesii) kraft pulp. A treatment with Irgazyme caused a much larger increase in brightness than did xylanase D. A double xylanase treatment with Irgazyme, before and after peroxide bleaching, resulted in the highest final brightness. Alkaline extraction increased the brightness of Douglas-fir brownstock. Treatment with Irgazyme released more lignin and carbohydrates than did xylanase D. The molecular mass of the lignin extracted from Irgazyme-treated brownstock was much larger than that from the control pulp. The lignin-like macromolecules directly solubilized from peroxide bleached pulps were substantially larger than those solubilized from the brownstock, irrespective of whether they were produced during xylanase or control treatments. This indicates that different kinds of materials were solubilized when a xylanase treatment was applied at different points in the bleaching sequence and raises concerns about the role of lignin entrapment in the mechanism by which xylanase enhances peroxide bleaching.

Análise de fragmentos arqueológicos: um diálogo entre a Química, Biologia e Arqueologia

Dissertação de mestrado em Técnicas de Caracterização e Análise Química

Determinação de psicofármacos em cabelo humano por técnicas cromatográficas hifenadas: um método alternativo não-invasivo

Dissertação de mestrado em Técnicas de Caracterização e Análise Química

Implementação e validação de um método analítico para a determinação de sucralose (E955) em refrigerantes e cervejas por HPLC-ELSD

Dissertação de mestrado em Técnicas de Caraterização e Análise Química

Left Ventricular Hypertrophy Evaluation in Obese Hypertensive Patients: Effect of Left Ventricular Mass Index Criteria

PURPOSE: To evaluate left ventricular mass (LVM) index in hypertensive and normotensive obese individuals. METHODS: Using M mode echocardiography, 544 essential hypertensive and 106 normotensive patients were evaluated, and LVM was indexed for body surface area (LVM/BSA) and for height² (LVM/h²). The 2 indexes were then compared in both populations, in subgroups stratified according to body mass index (BMI): <27; 27-30; > or = 30kg/m². RESULTS: The BSA index does not allow identification of significant differences between BMI subgroups. Indexing by height² provides significantly increased values for high BMI subgroups in normotensive and hypertensive populations. CONCLUSION: Left ventricular hypertrophy (LVH) has been underestimated in the obese with the use of LVM/BSA because this index considers obesity as a physiological variable. Indexing by height² allows differences between BMI subgroups to become apparent and seems to be more appropriate for detecting LVH in obese populations.

Development of nanohydrogels as a vehicle for the release of bioactive compounds in food matrices

PhD in Chemical and Biological Engineering

Body Mass Index: A Scientific Evidence-Based Inquiry

OBJECTVE: To objectively and critically assess body mass index and to propose alternatives for relating body weight and height that are evidence-based and that eliminate or reduce the limitations of the body mass index. METHODS: To analyze the relations involving weight and height, we used 2 databases as follows: 1) children and adolescents from Brazil, the United States, and Switzerland; and 2) 538 university students. We performed mathematical simulations with height data ranging from 115 to 190 cm and weight data ranging from 25 to 105 kg. We selected 3 methods to analyze the relation of weight and height as follows: body mass index - weight (kg)/height (m²); reciprocal of the ponderal index - height (cm)/weight1/3 (kg); and ectomorphy. Using the normal range from 20 to 25 kg/m² for the body mass index in the reference height of 170 cm, we identified the corresponding ranges of 41 to 44 cm/kg1/3 for the reciprocal of the ponderal index, and of 1.45 to 3.60 for ectomorphy. RESULTS: The mathematical simulations showed a strong association among the 3 methods with an absolute concordance to a height of 170 cm, but with a tendency towards discrepancy in the normal ranges, which had already been observed for the heights of 165 and 175 cm. This made the direct convertibility between the indices unfeasible. The reciprocal of the ponderal index and ectomorphy with their cut points comprised a larger age range in children and adolescents and a wider and more central range in the university students, both for the reported (current) and desired weights. CONCLUSION: The reciprocal of the ponderal index and ectomorphy are stronger and are more mathematically logical than body mass index; in addition, they may be applied with the same cut points for normal from the age of 5 ½ years on.

Influence of skeletal muscle mass on ventilatory and hemodynamic variables during exercise in patients with chronic heart failure

OBJECTIVE: To assess the influence of skeletal muscle mass on ventilatory and hemodynamic variables during exercise in patients with chronic heart failure (CHF). METHODS: Twenty-five male patients underwent maximum cardiopulmonary exercise testing on a treadmill with a ramp protocol and measurement of the skeletal muscle mass of their thighs by using magnetic resonance imaging. The clinically stable, noncachectic patients were assessed and compared with 14 healthy individuals (S) paired by age and body mass index, who underwent the same examinations. RESULTS: Similar values of skeletal muscle mass were found in both groups (CHF group: 3863 ± 874 g; S group: 3743 ± 540 g; p = 0.32). Significant correlations of oxygen consumption in the anaerobic threshold (CHF: r = 0.39; P= 0.02 and S: r = 0.14; P = 0.31) and of oxygen pulse also in the anaerobic threshold (CHF: r = 0.49; P = 0.01 and S: r =0.12; P = 0.36) were found only in the group of patients with chronic heart failure. CONCLUSION: The results obtained indicate that skeletal muscle mass may influence the capacity of patients with CHF to withstand submaximal effort, due to limitations in their physical condition, even maintaining a value similar to that of healthy individuals. This suggests qualitative changes in the musculature.

Ganadería de precisión 1. Desarrollo de un sistema electrónico para construir matrices genealógicas aplicables al mejoramiento animal en diversas especies (Programa: sustentabilidad productiva de pequeños rumiantes en áreas desfavorables - supprad).

El mejoramiento animal tiene como función modificar genéticamente una población animal con un fin económico determinado. Las herramientas que se utilizan son: evaluación genética, selección y/o esquemas de apareamiento. La evaluación genética se realiza obteniendo información (marcadores) que predice el genotipo del animal que no es registrable directamente. Estos marcadores son el fenotipo o información física del animal y algún predictor del genotipo que puede ser un marcador químico o la relación de parentesco con otro animal (padre-madre, hermanos, progenie). El uso de los marcadores genéticos para determinar la genealogía está en desarrollo actualmente pero resulta extremadamente costoso para ciertos niveles de producción. Por otra parte, los dispositivos electrónicos ensayados hasta el momento no han resultado muy eficientes, por lo tanto se propone para este proyecto desarrollar un sistema electrónico de identificación animal que permita construir matrices de genealogía para la evaluación genética de reproductores de las diferentes especies en condiciones de producción extensivas. El proyecto se desarrollará entre especialistas en Mejoramiento Animal e Ingenieros Electrónicos, en un aporte multidisciplinario a la solución del problema de establecer la filiación a través del armado del par macho-hembra durante el servicio natural de las distintas especies y el par madre-cría en algún momento luego del parto. La solución propuesta consiste en la implementación de dispositivos electrónicos activos (microcontrolador y tranceptor de radiofrecuencia alimentados a batería) dispuestos como nodos de una red de sensores inalambricos. Estos dispositivos serán colocados a través de collares en los animales tomados como objeto de estudio y mediante el continuo intercambio de informacion entre ellos se generará una matriz de datos que permita a posterior establecer la relación que existe entre ellos a traves de la determinacion de la periodicidad de sus cercanías relativas.

Antioxidantes de matrices vegetales regionales concentrados con tecnologías de alto vacío

El objetivo general de este proyecto es el de estudiar la recuperación de nutracéuticos y antioxidantes a partir de orégano y romero, tanto evitando su perdida durante el procesamiento como mediante su extracción y concentración utilizando tecnologías eficientes y no contaminantes. Se fraccionarán los aceites esenciales y oleoresinas, por destilación molecular para obtener productos de alto contenido en nutracéuticos. Se estudiará la extracción-separación de nutracéuticos de un modo teórico-experimental. Se realizarán determinaciones experimentales de cada una de las operaciones, para distintos valores de las variables operativas más relevantes a efectos de estudiar su influencia y lograr la optimización de las operaciones. Se utilizarán los modelos de Redes Neuronales Artificiales para el modelado y optimizacion de las operaciones implementadas.

Procesos oxidativos en matrices alimentarias complejas. Oxidative processes in complex food matrices.

Las reacciones bioquímicas que ocurren como consecuencia del tratamiento y almacenamiento de los alimentos, mejoran la seguridad alimentaria, las propiedades sensoriales y la vida útil. Sin embargo, el tratamiento térmico, la exposición a la luz y el oxígeno pueden causar daño oxidativo a los lípidos y proteínas. Los procesos oxidativos de matrices complejas tienen características distintivas que no se manifiestan cuando los componentes son sometidos a oxidación individualmente. La hipótesis de trabajo es que la oxidación de proteínas en matrices alimentarias complejas altera la estructura y las propiedades funcionales de las proteínas y, que las modificaciones que se producen varían según las condiciones de procesamiento y de la composición química del alimento. Nuestros estudios intentan demostrar que el estado oxidativo de las proteínas de un alimento es un parámetro importante para la evaluación de las propiedades funcionales, sensoriales y nutricionales de un producto lácteo. El objetivo general del proyecto es el estudio de los procesos de oxidación de matrices alimentarias complejas (la leche, miel) y su relación con distintos procesos y materiales utilizados en la industria. Es decir, nos proponemos estudiar las consecuencias funcionales y biológicas (calidad nutricional, coagulación) de la oxidación proteica en modelos experimentales “in vitro” y en productos comerciales. 1. Estudiar los fenómenos de peroxidación proteica en leche entera y descremada sometida a los distintos procesos tecnológicos de la producción de leche y queso a escala laboratorio. Se realizarán las mismas experiencias con albúmina sérica y con proteínas aisladas de suero de leche para comparar diferencias entre una matriz compleja y una simple. 2. Determinar la relación entre oxidación y composición proteica de la leche, y los cambios en las fracciones proteicas aisladas (caseínas y beta-lactoglobulina). 3. Analizar el impacto de los procesos tecnológicos a nivel de producción primaria (composición proteica y estado de oxidación) en los indicadores de inflamación (contenido de células somáticas y proteína C Reactiva) y de estado redox (capacidad antioxidante de los productos lácteos y nivel de carbonilos de proteinas). 4. Comparar las características de composición química y el estado de oxidación de leche provenientes de las tres regiones (Buenos Aires, Santa Fe y Córdoba) que conforman la cuenca láctea Argentina. Este objetivo se realizará conjuntamente con los integrantes de nuestro grupo de investigación que trabajan en el Laboratorio de Control de Calidad de la Escuela Superior de Lechería. 5. Determinar los metabolitos secundarios de mieles uniflorales propuestos como responsables de la capacidad antioxidante de estas (polifenoles) y como indicadores de su origen botánico. 6. Valorar la capacidad antioxidante total de mieles uniflorales. 7. Validar los métodos analíticos y semicuantitativos utilizados y a utilizar en el presente proyecto teniendo en cuenta lo efectos de matrices típico de los fluidos biológicos y las mezclas. El estudio de las modificaciones oxidativas de matrices complejas es un tema que es importante tanto desde el punto de vista del conocimiento básico como del aplicado. Nosotros creemos que el presente proyecto aportará conocimiento sobre las características de las vías oxidativas de proteínas en matrices complejas y que podrá ser utilizado para diseñar estrategias productivas tendientes a disminuir el deterioro de la calidad de la leche debido a la exposición a energía radiante. Parte de la experiencia ganada por el grupo ha sido ya volcada a subsanar dificultades y problemas de oxidación y deterioro de la calidad de alimentos. Además, se contribuirá a discernir la paradoja que existe en el área sobre las propiedades oxidantes/antioxidantes de los polifenoles y la relación entre estas y el estado oxidativo de un alimento. The biochemical reactions that occur as a result of food treatment and storage, improve food security, sensory properties and shelf life. Heat treatment, exposure to light and oxygen can cause oxidative damage to lipids and proteins. Oxidative processes in complex matrices display distinctive features that do not appear when the components are individually subjected to oxidation. The hypothesis is that protein oxidation in complex food matrices alters the structure and functional properties of proteins and that the modifications vary according to process conditions and food composition. The main goal is to study oxidation of complex food matrices (milk, honey) with different processes and materials used in the industry. The specific aims are: 1. To study protein oxidation in whole milk and skim subject to various technological processes. The same experiences will be done with serum albumin and isolated whey proteins to compare complex and simple matrices. 2. To determine the relationship between oxidation and milk protein composition, and changes in casein and beta-lactoglobulin. 3. Analyze the impact of technological processes at the level of primary production on markers of inflammation and redox (antioxidant capacity and protein carbonyls). 4. Compare characteristics of chemical composition and oxidation state of milk. 5. Determine secondary metabolites of honey responsible for the antioxidant capacity of these. 6. To evaluate the total antioxidant capacity unifloral honey. This project will provide knowledge about characteristics of oxidative pathways of proteins in complex matrices that can be used to design production strategies aimed at reduce the deterioration of milk quality. Also, it would help to discern the paradox that exists on the oxidants/antioxidants properties of polyphenols and the relationship between these and the oxidative status of a food.

GANADERÍA DE PRECISIÓN. DESARROLLO DE UN SISTEMA ELECTRÓNICO PARA CONSTRUIR MATRICES GENEALÓGICAS APLICABLES AL MEJORAMIENTO ANIMAL EN DIVERSAS ESPECIES.

El mejoramiento animal tiene como función modificar genéticamente una población animal con un fin económico determinado. Las herramientas que se utilizan son: evaluación genética, selección y/o esquemas de apareamiento. La evaluación genética se realiza obteniendo información (marcadores) que predice el genotipo del animal que no es registrable directamente. Estos marcadores son el fenotipo o información física del animal y algún predictor del genotipo que puede ser un marcador químico o la relación de parentesco con otro animal. El uso de los marcadores genéticos para determinar la genealogía está en desarrollo actualmente pero resulta extremadamente costoso para ciertos niveles de producción. Los dispositivos electrónicos ensayados hasta el momento no han resultado muy eficientes, por lo tanto se propone para este proyecto desarrollar un sistema electrónico de identificación animal que permita construir matrices de genealogía para la evaluación genética de reproductores de las diferentes especies en condiciones de producción extensivas. El proyecto se desarrollará entre especialistas en mejoramiento animal e ingenieros electrónicos, en un aporte multidisciplinario a la solución del problema de establecer la filiación a través del armado del para macho-hembra durante el servicio natural de las distintas especies y el par madre-cría en algún momento luego del parto. La solución propuesta consiste en la implementación de dispositivos electrónicos activos (microcontrolador y tranceptor de radiofrecuencia alimentados a batería) dispuestos como nodos de una red de sensores inalambricos. Estos dispositivos serán colocados en los animales tomados como objeto de estudio, y mediante el continuo intercambio de informacion entre ellos se genera una matriz de datos que permita a posterior establecer la relación que existe entre ellos a traves de la determinacion de la periodicidad de sus cercanias relativas.