Primary bioreceptivity of limestones from the mediterranean basin to phototrophic microorganisms

Dissertação apresentada para a obtenção do grau de Doutor em Conservação e Restauro pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

The influence of inherent properties of building limestones on their bioreceptivity to phototrophic microorganisms

Annals of Microbiology, 59 (4) 705-713 (2009)

Relative Frequency of Nosocomial Microorganisms at Unicamp University Hospital from 1987 to 1994

The frequency of microorganisms identified in nosocomial infections at Unicamp University Hospital from 1987 to 1994 was analysed. The most common microorganism was S. aureus (20.9%), which was found in surgical wound, bloodstream and arterial-venous infections. In urinary tract infections (UTI), gram-negative rods (56.5%) and yeasts (9%) predominated. A. baumannii isolates were observed to have increased in the last three years. There was a gradual increase in the frequency of coagulase-negative staphylococci and A. baumannii in bloodstream infections but there wasn’t any change in Candida sp

IDENTIFICATION OF Pseudomonas spp. AS AMOEBA-RESISTANT MICROORGANISMS IN ISOLATES OF Acanthamoeba

Acanthamoeba is a “Trojan horse” of the microbial world. The aim of this study was to identify the presence of Pseudomonas as an amoeba-resistant microorganism in 12 isolates of Acanthamoeba. All isolates showed the genus Pseudomonas spp. as amoeba-resistant microorganisms. Thus, one can see that the Acanthamoeba isolates studied are hosts of Pseudomonas.

Synthesis of new enzyme stabilisers inspired by compatible solutes of hyperthermophilic microorganisms

Dissertation presented to obtain the Ph.D degree in Engineering Sciences and Technology.

Public service culture collections of microorganisms: are they important for biotechnology?

Microbiology as a scientific discipline recognised the need to preserve microorganisms for scientific studies establishing from its very beginning research culture collections (CC). Later on, to better serve different scientific fields and bioindustries with the increasing number of strains of scientific, medical, ecological and biotechnological importance public service CC were established with the specific aims to support their user communities. Currently, the more developed public service CC are recognised as microBiological Resources Centres (mBRC). mBRC are considered to be one of the key elements for sustainable international scientific infrastructure, which is necessary to underpin successful delivery of the benefits of biotechnology, whether within the health sector, the industrial sector or other sectors, and in turn ensure that these advances help drive economic growth. In more detail, mBRCs are defined by Organisation for Economic Co-operation and Development (OECD) as service providers and repositories of the living cells, genomes of organisms, and information relating to heredity and functions of biological systems. mBRCs contain collections of culturable organisms (e.g., microorganisms, plant, animal cells), replicable parts of these (e.g. genomes, plasmids, virus, cDNAs), viable but not yet culturable organisms, cells and tissues, as well as database containing molecular, physiological and structural information relevant to these collections and related bioinformatics. Thus mBRCs are fundamental to harnessing and preserving the world’s microbial biodiversity and genetic resources and serve as an essential element of the infrastructure for research and development. mBRCs serve a multitude of functions and assume a range of shapes and forms. Some are large national centres performing a comprehensive role providing access to diverse organisms. Other centres play much narrower, yet important, roles supplying limited but crucial specialised resources. In the era of the knowledge-based bio-economy mBRCs are recognised as vital element to underpinning the biotechnology.

Antimicrobial activity of honeys from two stingless honeybee species and Apis mellifera (Hymenoptera: Apidae) against pathogenic microorganisms

Honeys are described possessing different properties including antimicrobial. Many studies have presented this activity of honeys produced by Apis mellifera bees, however studies including activities of stingless bees honeys are scarce. The aim of this study was to compare the antimicrobial activity of honeys collected in the Amazonas State from Melipona compressipes, Melipona seminigra and Apis mellifera against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Chromobacterium violaceum, and Candida albicans. Minimum inhibitory concentrations were determined using the agar dilution method with Müller-Hinton agar (for bacteria) or Saboraud agar (for yeast). Staphylococcus aureus and E. faecalis were inhibited by all honeys at concentrations below 12%, while E. coli and C. violaceum were inhibited by stingless bee honeys at concentrations between 10 and 20%. A. mellifera honey inhibited E. coli at a concentration of 7% and Candida violaceum at 0.7%. C. albicans were inhibited only with honey concentrations between 30 and 40%. All examined honey had antimicrobial activity against the tested pathogens, thus serving as potential antimicrobial agents for several therapeutic approaches.

Biosurfactants produced by marine microorganisms with therapeutic applications

Marine microorganisms possess unique metabolic and physiological features and are an important source of new biomolecules, such as biosurfactants. Some of these surface-active compounds synthesized by marine microorganisms exhibit antimicrobial, anti-adhesive and anti-biofilm activity against a broad spectrum of human pathogens (including multi-drug resistant pathogens), and could be used instead of the existing drugs to treat infections caused by them. In other cases, these biosurfactants show anti-cancer activity, which could be envisaged as an alternative to conventional therapies. However, marine biosurfactants have not been widely explored, mainly due to the difficulties associated with the isolation and growth of their producing microorganisms. Culture-independent techniques (metagenomics) constitute a promising approach to study the genetic resources of otherwise inaccessible marine microorganisms without the requirement of culturing them, and can contribute to discover novel biosurfactants with significant biological activities. This paper reviews the most relevant biosurfactants produced by marine microorganisms with potential therapeutic applications and discusses future perspectives and opportunities to discover novel molecules from marine environments.

Microbial respiration with chlorine oxyanions: diversity and physiological and biochemical properties of chlorate- and perchlorate-reducing microorganisms

Chlorine oxyanions are valuable electron acceptors for microorganisms. Recent findings have shed light on the natural formation of chlorine oxyanions in the environment. These suggest a permanent introduction of respective compounds on Earth, long before their anthropogenic manufacture. Microorganisms that are able to grow by the reduction of chlorate and perchlorate are affiliated with phylogenetically diverse lineages, spanning from the Proteobacteria to the Firmicutes and archaeal microorganisms. Microbial reduction of chlorine oxyanions can be found in diverse environments and different environmental conditions (temperature, salinities, pH). It commonly involves the enzymes perchlorate reductase (Pcr) or chlorate reductase (Clr) and chlorite dismutase (Cld). Horizontal gene transfer seems to play an important role for the acquisition of functional genes. Novel and efficient Clds were isolated from microorganisms incapable of growing on chlorine oxyanions. Archaea seem to use a periplasmic Nar-type reductase (pNar) for perchlorate reduction and lack a functional Cld. Chlorite is possibly eliminated by alternative (abiotic) reactions. This was already demonstrated for Archaeoglobus fulgidus, which uses reduced sulfur compounds to detoxify chlorite. A broad biochemical diversity of the trait, its environmental dispersal, and the occurrence of relevant enzymes in diverse lineages may indicate early adaptations of life toward chlorine oxyanions on Earth.

Zeolite-based nanomaterials for the control of opportunistic pathogenic microorganisms

Dissertação de mestrado em Applied Biochemistry (área de especialização em Biomedicine)

Desarrollo de fotosensibilizadores con aplicaciones en la inactivación fotodinámica de microorganismos. Development of photosensitizer with applications in the photodynamic inactivation of microorganisms.

El plan propone desarrollar nuevas agentes fotosensibilizadores derivados de macrociclos pirrólicos con aplicaciones en la inactivación fotodinámica (PDI) de microorganismos. La propuesta abarca el desarrollo de procedimientos apropiados para la síntesis de compuestos derivados de porfirinas, subftalocianinas y ftalocianinas sustituidas en la periferia por grupos que permitan aumentar la actividad biológica. Con la finalidad de incrementar la incorporación intracelular y la actividad fotodinámica se evaluarán sensibilizadores con distinta distribución y número de cargas, en los cuales se ha incrementado el carácter anfifílico por la presencia de grupos lipofílicos y catiónicos. La combinación de un fotosensibilizador con un compuesto antifúngico está diseñada para aumentar la eficiencia en la inactivación de hongos. También serán evaluadas superficies antimicrobianas recubiertas con una película de fotosensibilizadores. En primera instancia, la actividad fotodinámica de los nuevos agentes fototerapéuticos serán evaluados en sistemas biomiméticos conteniendo sustratos biológicamente activos. Los estudios in vitro serán realizados en cultivos de bacterias y levaduras. Esta aplicación presenta considerable importancia en la inactivación de microorganismos patógenos que crecen in vivo en un foco localizado de infección, en la desinfección de fluidos biológicos y aguas contaminadas con microbios resistentes.

Biorremediación de cromo y fenol mediante la aplicación de microorganismos nativos de la Provincia de Córdoba Bioremediation of chromium and phenol by the application of native microorganisms from Córdoba Province

La contaminación ambiental por metales pesados como el cromo y por compuestos orgánicos como los fenoles es un grave problema a nivel mundial debido a su toxicidad y a sus efectos adversos sobre los seres humanos, la flora y la fauna, tanto por su acumulación en la cadena alimentaria como por su continua persistencia en el medio ambiente. En un estudio preliminar, efectuado por nuestro laboratorio, se han detectado elevados niveles de estos contaminantes en sedimentos y efluentes en zonas industriales del sur de la provincia de Córdoba, lo cual plantea la necesidad de removerlos. Entre las tecnologías disponibles, la biorremediación, que se basa en el uso de sistemas biológicos, como los microorganismos, para la detoxificación y la degradación de contaminantes, se presenta como una alternativa probablemente más efectiva y de menor costo que las técnicas convencionales. Sin embargo, la aplicación de esta tecnología depende en gran parte de la influencia de las características particulares y específicas de la zona a remediar. En consecuencia, en primer lugar se caracterizará la zona de muestreo y se aislarán e identificarán microorganismos nativos de la región, tolerantes a cromo y fenol, a partir de muestras de suelo, agua y sedimentos, ya que podrían constituir una adecuada herramienta biotecnológica, mejor adaptada al sitio a tratar. Posteriormente se estudiará la biorremediación de Cr y fenol utilizando dichos microorganismos, analizando su capacidad para biotransformar, bioacumular o bioadsorber a estos contaminantes, y se determinarán las condiciones óptimas para el tratamiento. Se analizarán los posibles mecanismos fisiológicos, bioquímicos y moleculares involucrados en la remediación, que constituye una etapa crucial para el diseño de una estrategia adecuada y eficiente. Finalmente, se aplicará esta tecnología a escala reactor, como una primera aproximación al tratamiento a mayor escala. De esta manera se espera reducir los niveles de estos contaminantes y así minimizar el impacto ambiental que ellos producen en suelos y acuíferos. A futuro, la utilización de los microorganismos seleccionados, de manera individual o formando consorcios, para el tratamiento de efluentes industriales previa liberación al medio ambiente, o su uso en bioaumento, constituirían posibles alternativas de aplicación. Los principales impactos científico-tecnológicos del proyecto serán: (a) la generación de una nueva tecnología biológica de decontaminación de cromo y fenol, intentando presentar soluciones frente a una problemática ambiental que afecta a nuestra región, pero que además es común a la mayoría de los países, (b) la formación de nuevos recursos humanos en el área y (c) el trabajo en colaboración con otros grupos de investigación que se destacan en el área de biotecnología ambiental. Environmental pollution produced by heavy metals, such as chromium and organic compounds like phenolics is a serious global problem due to their toxicity, their adverse effects on human life, plants and animals, their accumulation in the food chains and also by their persistance in the environment. In a previous study performed in our laboratory, high levels of these pollutants were detected in sediments and effluents from industrial zones of the south of Cordoba Province, which determine the need to remove them. Among various technologies, bioremediation which is based on the use of biological systems, such as microorganisms, to detoxify and to degrade contaminants, is probably the most effective alternative, and it is less expensive than other conventional technologies. However, the application of this technology depends on the influence of the particular and specific characteristics of the zone to be remediate. As a consecuence, at the first time, the zone of sampling will be characterized and then, native microorganisms, tolerant to chromium and phenol, will be isolated from soils, water and sediments and identificated. These microorganisms would be an adequate biotechnological tool, more adapted to the conditions of the site to be remediate than other ones. Then, the ability of these selected microorganisms to biotransform, bioaccumulate or biosorbe chromium and phenol will be studied and the optimal conditions for the treatment will be determined. The possible physiological, biochemical and molecular mechanisms involved in bioremediation will be also analized, because this is a crucial step in the design of an adequate and efficient remediation strategy. Finally, this technology will be applied in a reactor, as an approximation to the treatment at a major scale. A reduction in the levels of these pollutants will be expected, to minimize their environmental impact on soils and aquifers.

Infection by microorganisms on soybean seeds obtained from plants treated with growth regulators

This research, deals with the effects of exogenous growth regulators on infection by microorganisms on soybean (Glycine max cv. Davis) seeds. To study the influence of the chemicals, soybean plants were sprayed with gibberellic acid (GA) 100 ppm, (2-chloroethyl) trimethylammonium chloride (CCC) 2,000 ppm, succinic acid-2,2-dimethy1hydrazide (SADH) 4,000 ppm, indolylacetic acid (IAA) 100 ppm, 2,3,5-triiodobenzoic acid (TIBA) 20 ppm (three applications), and Agrostemin (1g/10 ml/ 3 1). Application of growth regulators did not affect infect ion by microorganisms on soybean seeds. The prominent fungus isolated was Phomopsis sojae. Alternaria and Fusarium spp. were isolated from seeds. The presence of a bacterium on the seeds was observed. The delay in harvest and high humidity increased the number of seeds from which Phomopsis was recovered.