Biodegradation of Thermoplastic and hermosetting Polyesters from Z-Protected Glutamic Acid

In a previous article,1 the development and molecular characterization of three polyesters from N-carbobenzyloxy-L-glutamic acid (ZGluOH) were reported. The polymers were a linear, heterochain polyester (ZGluOH and ethylene glycol), a crosslinked heterochain polyester (ZGluOH and diglycidyl ether of 1,4-butanediol), and a crosslinked, heterochain aromatic polyester (ZGluOH and diglycidyl ether of bisphenol A). In this manuscript, results of biodegradation studies are reported. The three polymers hydrolyzed to low molecular weight oligomers similar to the monomers with lipase. When exposed to a mixed culture of micro-organisms, the first two resins degraded to biomass and respiratory gases. The crosslinked heterochain aromatic polyester resisted microbial degradation.

Ocorrência de fungos negros em ambientes domésticos

Black fungi are able to adapt to extreme environmental conditions, such as: high temperatures, the presence of toxic chemical substances and lack of nutrients. Besides, they are also potential pathogens to humans. The natural environment of many black fungi is still unknown and some studies are being conducted to evaluate the biodiversity of this group and their different habitats. This study aimed to isolate black fungi in domestic environments and facilities, such as toothbrushes, fridge sealing rubbers, bathroom strainers and divisions, windows, wall tiles and bath sponge. For the collection, material surfaces were scratched with a scalpel and the resulting fragments were sewed in Mycosel agar (DifcoTM), supplemented with actidione to inhibit the growth of highly-sporulating fungi. Plates were incubated at 25ºC for three weeks. The 46 isolated fungi were maintained on MA2% slants at 8ºC and cryopreserved at -80ºC. Fungal identification was performed through the analysis of macro and microscopic features and ITS rDNA sequencing. The following black fungi taxa were found: Ascomycota sp., Cladosporium spp., Dothideomycete sp., Exophiala alcalophila, Ochroconis mirabilis and Rhinocladiella atrovirens. Non-melanized fungi were also found, such as Geosmithia sp., Penicillium sp. and Rhodotorula mucilaginosa. The temperature tests showed that isolated black fungi were not able to grow at 37°C, however, this temperature proved to be fungistatic to 43% of them. According to literature, all black fungi isolated in this study are opportunistic pathogens and additional studies are necessary to evaluate the risk that these micro-organisms offer to health, once they were isolated from domestic environments

Production and characterization of poly-(3-hydroxybutyrate) from recombinant Escherichia coli grown on cheap renewable carbon substrates

Although the biopolymer poly-(3-hydroxybutyrate), P[3HB], presents physicochemical properties that make it an alternative material to conventional plastics, its biotechnological production is quite expensive. As carbon substrates contribute greatly to P[3HB] production cost, the utilization of a cheaper carbon substrate and less demanding micro-organisms should decrease its cost. In the present study a 23 factorial experimental design was applied, aiming to evaluate the effects of using hydrolysed corn starch (HCS) and soybean oil (SBO) as carbon substrates, and cheese whey (CW) supplementation in the mineral medium (MM) on the responses, cell dried weigh (DCW), percentage P[3HB] and mass P[3HB] by recombinant Escherichia coli strains JM101 and DH10B, containing the P[3HB] synthase genes from Cupriavidus necator (ex-Ralstonia eutropha). The analysis of effects indicated that the substrates and the supplement and their interactions had positive effect on CDW. Statistically generated equations showed that, at the highest concentrations of HCS, SO and CW, theoretically it should be possible to produce about 2 g L(1) DCW, accumulating 50% P[3HB], in both strains. To complement this study, the strain that presented the best results was cultivated in MM added to HCS, SBO and CW ( in best composition observed) and complex medium (CM) to compare the obtained P[3HB] in terms of physicochemical parameters. The obtained results showed that the P[3HB] production in MM (1.29 g L(-1)) was approximately 20% lower than in CM (1.63 g L(-1)); however, this difference can be compensated by the lower cost of the MM achieved by the use of cheap renewable carbon sources. Moreover, using differential scanning calorimetry and thermogravimetry analyses, it was observed that the polymer produced in MM was the one which presented physicochemical properties (Tg and Tf) that were more similar to those found in the literature for P[3HB].

Brazilian research on extremophiles in the context of astrobiology

Extremophiles are organisms adapted to grow at extreme ranges of environmental variables, such as high or low temperatures, acid or alkaline medium, high salt concentration, high pressures and so forth. Most extremophiles are micro-organisms that belong to the Archaea and Bacteria domains, and are widely spread across the world, which include the polar regions, volcanoes, deserts, deep oceanic sediments, hydrothermal vents, hypersaline lakes, acid and alkaline water bodies, and other extreme environments considered hostile to human life. Despite the tropical climate, Brazil has a wide range of ecosystems which include some permanent or seasonally extreme environments. For example, the Cerrado is a biome with very low soil pH with high Al+3 concentration, the mangroves in the Brazilian coast are anaerobic and saline, Pantanal has thousands of alkaline-saline lakes, the Caatinga arid and hot soils and the deep sea sediments in the Brazilian ocean shelf. These environments harbour extremophilic organisms that, coupled with the high natural biodiversity in Brazil, could be explored for different purposes. However, only a few projects in Brazil intended to study the extremophiles. In the frame of astrobiology, for example, these organisms could provide important models for defining the limits of life and hypothesize about life outside Earth. Brazilian microbiologists have, however, studied the extremophilic micro-organisms inhabiting non-Brazilian environments, such as the Antarctic continent. The experience and previous results obtained from the Brazilian Antarctic Program (PROANTAR) provide important results that are directly related to astrobiology. This article is a brief synopsis of the Brazilian experience in researching extremophiles, indicating the most important results related to astrobiology and some future perspectives in this area. Received 29 February 2012, accepted 25 May 2012, first published online 11 July 2012

Effects of Aspergillus spp. exogenous fibrolytic enzymes on in vitro fermentation of tropical forages

BACKGROUND: Cellulose and hemicellulose are quantitatively the most important structural carbohydrates present in ruminant diets. Rumen micro-organisms produce enzymes that catalyse their hydrolysis, but the complex network formed by structural carbohydrates and lignin reduces their digestibility and restricts efficient utilisation of feeds by ruminants. This study aimed to produce two enzymatic extracts, apply them in ruminant diets to determine the best levels for ruminal digestibility and evaluate their effects on in vitro digestibility. RESULTS: In experiment 1 a two-stage in vitro technique was used to examine the effects of different enzymatic levels of Aspergillus japonicus and Aspergillus terricola on tropical forages. Enzyme addition had minor effects on corn silage at the highest enzymatic level. In experiment 2 an in vitro gas production (GP) technique was applied to determine apparent in vitro organic matter digestibility and metabolisable energy. The addition of enzymes in GP showed interesting results. Good data were obtained using sugar cane and Tifton-85 hay supplemented with extracts of A. japonicus and A. terricola respectively. CONCLUSION: Overall, the study suggests that addition of crude extracts containing exogenous fibrolytic enzymes to ruminant diets enhances the effective utilisation of ruminant feedstuffs such as forages. Copyright (c) 2012 Society of Chemical Industry

Phylogenetische Charakterisierung von Mikroorganismen aus dem Intestinaltrakt von Insekten

Zusammenfassung Die komplexe Lebensgemeinschaft des Termitendarms fasziniert die Biologen schon seit langem. Es ist bekannt, dass Termiten ihre Nahrung mit Hilfe von symbiontischen Bakterien und Protozoen verdauen können. Ohne ihre Symbionten würden sie verhungern. Das Zusammenspiel von Termiten und darmbewohnenden Mikroorganismen, zu denen Flagellaten, Bakterien, Archaebakterien und Hefen gehören, ist trotz moderner Untersuchungstechniken keineswegs vollständig aufgeklärt. In der vorliegenden Arbeit wurden:1) Einige kultivierte und nicht-kultivierte Bakterien charakterisiert, die an der Darmwand von Mastotermes darwiniensis lokalisiert sind. Die Darmwandbakterien wurden entweder nach Kultivierung oder direkt von der Darmwand für die Analyse der 16S rDNA verwendet. Die Sequenzierung erfolgte entweder nach DGGE oder nach Klonierung der PCR-Produkte. Die identifizierten Bakterien kann man in 7 Gruppen teilen:1: Gram-positive Bakterien mit hohem GC-Gehalt 2: Gram-positive Bakterien mit niedrigem GC-Gehalt 3: Fusobakterien-ähnliche Bakterien 4: ß-Proteobakterien5: Verrucomicrobien6: Bacteroides-ähnliche Bakterien7: Methanogene Bakterien 2) Aufgrund des Vorhandenseins des Coenzyms Deazaflavin-Derivats F420, kann man Methanbakterien mikroskopisch identifizieren und von anderen Bakterien unterscheiden, weil Methanbakterien im kurzwelligen Blaulicht blaugrün aufleuchten. Untersuchungen haben gezeigt, dass mindestens zwei Morphotypen von Methanbakterien an der Darmwand von M. darwiniensis vorkommen. Sie wurden auch über 16S rDNA Sequenzanalyse identifiziert. Ihre Lokalisierung an der Darmwand wurde durch Fluoreszenz-in-situ-Hybridsierung mit spezifischen Oligonukleotiden nachgewiesen. Schließlich konnte gezeigt werden, dass pro Gramm Termite 2,6 µg Methan pro Stunde produziert werden. 3) Bis jetzt wurden aus verschiedenen Termiten sulfatreduzierende Bakterien (SRB) isoliert. Deshalb wurde in dieser Arbeit die Verbreitung der SRB in verschiedenen Insekten untersucht. Insgesamt wurden zwei Sequenzen aus Libellenlarven (FSBO4 und FSBRO2), drei Sequenzen aus Zuckmückenlarven (FSCI, FSCII und FSC4), eine Sequenz aus Rosenkäfern (FSPa4-5) und ebenfalls eine Sequenz aus Eintagsfliegenlarven (FSB6) identifiziert. Alle identifizierten Bakterien ausser Klon FSB6, gehören zur Gattung Desulfovibrio. Klon FSB6 gehört zu der Gram-positiven Gattung Desulfotomaculum.Außerdem wurde die Sulfatreduktionsrate der SRB im Darm von Rosenkäfern (Pachnoda marginata), Holz- bzw. Sulfat-gefütterten Termiten (Mastotermes darwiniensis) und einer Reinkultur von Desulfovibrio intestinalis gemessen. Dabei konnte gezeigt werden, dass die Aktivität pro Zelle in Holz-gefütterten Termite am höchsten ist (4,9 nmol/107 Bakterien x h).

Development of guidelines for microbiological control in microbrewery

Over the past 15 years the Italian brewing scene showed interesting changes, especially with regard to the creation of many breweries with an annual production of less than 10,000 hectoliters. The beers produced by microbreweries are very susceptible to attack by spoilage micro-organisms that cause the deterioration of beer quality characteristics. In addition, most of the microbreweries do not practice heat treatments of stabilization and do not carry out quality checks on the product. The high presence of beer spoilage bacteria is an economic problem for the brewing industry because it can damage the brand and it causes high costs of product retrieval. This thesis project was aimed to study the management of the production process in the Italian microbreweries within a production less than 10,000 hl. In particular, the annual production, type of plant, yeast management, process management, cleaning and sanitizing of a representative sample of microbreweries were investigated. Furthermore was made a collection of samples in order to identify, with simple methods, what are spoilage bacteria more present in the Italian craft beers. 21% of the beers analysed were positive at the presence of lactic acid bacteria. These analytical data show the importance of understanding what are the weak points of the production process that cause the development of spoilage bacteria. Finally, the thesis examined the actual production of two microbreweries in order to understand the process management that can promote the growth of spoilage bacteria in beer and production plant. The analysis of the data for the two case studies was helpful to understand what are the critical points where the microorganisms are most frequently in contact with the product. The hygiene practices are crucial to ensure the quality of the finished product, especially in the case of non-pasteurized beer.

Evolution of the structural and fracture design of the ISS payloads due to the new launchers, specifically applied to Biolab IEC-2 projects

The relatively young discipline of astronautics represents one of the scientifically most fascinating and technologically advanced achievements of our time. The human exploration in space does not offer only extraordinary research possibilities but also demands high requirements from man and technology. The space environment provides a lot of attractive experimental tools towards the understanding of fundamental mechanism in natural sciences. It has been shown that especially reduced gravity and elevated radiation, two distinctive factors in space, influence the behavior of biological systems significantly. For this reason one of the key objectives on board of an earth orbiting laboratory is the research in the field of life sciences, covering the broad range from botany, human physiology and crew health up to biotechnology. The Columbus Module is the only European low gravity platform that allows researchers to perform ambitious experiments in a continuous time frame up to several months. Biolab is part of the initial outfitting of the Columbus Laboratory; it is a multi-user facility supporting research in the field of biology, e.g. effect of microgravity and space radiation on cell cultures, micro-organisms, small plants and small invertebrates. The Biolab IEC are projects designed to work in the automatic part of Biolab. In this moment in the TO-53 department of Airbus Defence & Space (formerly Astrium) there are two experiments that are in phase C/D of the development and they are the subject of this thesis: CELLRAD and CYTOSKELETON. They will be launched in soft configuration, that means packed inside a block of foam that has the task to reduce the launch loads on the payload. Until 10 years ago the payloads which were launched in soft configuration were supposed to be structural safe by themselves and a specific structural analysis could be waived on them; with the opening of the launchers market to private companies (that are not under the direct control of the international space agencies), the requirements on the verifications of payloads are changed and they have become much more conservative. In 2012 a new random environment has been introduced due to the new Space-X launch specification that results to be particularly challenging for the soft launched payloads. The last ESA specification requires to perform structural analysis on the payload for combined loads (random vibration, quasi-steady acceleration and pressure). The aim of this thesis is to create FEM models able to reproduce the launch configuration and to verify that all the margins of safety are positive and to show how they change because of the new Space-X random environment. In case the results are negative, improved design solution are implemented. Based on the FEM result a study of the joins has been carried out and, when needed, a crack growth analysis has been performed.

The Legal Protection of Fundamental Human Values in Central and Eastern Europe and Modern Biotechnology: Towards European Harmonization

The protection of the fundamental human values (life, bodily integrity, human dignity, privacy) becomes imperative with the rapid progress in modern biotechnology, which can result in major alterations in the genetic make-up of organisms. It has become possible to insert human genes into pigs so that their internal organs coated in human proteins are more suitable for transplantation into humans (xenotransplantation), and micro-organisms that cam make insulin have been created, thus changing the genetic make-up of humans. At the end of the 1980s, the Central and Eastern European (CEE) countries either initiated new legislation or started to amend existing laws in this area (clinical testing of drugs, experiments on man, prenatal genetic diagnosis, legal protection of the embryo/foetus, etc.). The analysis here indicates that the CEE countries have not sufficiently adjusted their regulations to the findings of modern biotechnology, either because of the relatively short period they have had to do so, or because there are no definite answers to the questions which modern biotechnology has raised (ethical aspects of xenotransplantation, or of the use of live-aborted embryonic or foetal tissue in neuro-transplantation, etc.). In order to harmonise the existing regulations in CEE countries with respect to the EU and supranational contexts, two critical issues should be taken into consideration. The first is the necessity for CEE countries to recognise the place of humans within the achievements of modern biotechnology (a broader affirmation of the principle of autonomy, an explicit ban on the violation of the genetic identity of either born or unborn life, etc.). The second concerns the definition of the status of different biotechnological procedures and their permissibility (gene therapy, therapeutic genomes, xenotransplantation, etc.). The road towards such answers may be more easily identified once all CEE countries become members of the Council of Europe and express their wish to join the EU, which in turn presupposes taking over the entire body of EU legislation.

Conodonts from the Quadrant Shales of Montana

Conodonts have not been known to exist in Montana prior to the discovery of the specimens discussed and described in this paper. With this fact in mind, the author has endeavored to correlate the species of seven genera found in the Quadrant formation of Montana with similar micro-organisms from other North American areas.

Oceanography during TYDEMAN cruise DCM (DeepChlorMax) to the Equatorial Atlantic

The cruise with RV Tydeman was devoted to study permanently stratified plankton systems in the (sub)tropical ocean, which are characterised by a deep chlorophyll peak between 80 and 150 m. To minimise lateral effects by horizontal transport of nutrients and organic matter from river outflow and upwelling regions, stations were selected in the middle of the North Atlantic Ocean between the continents of America and Africa. (5 - 35° N and 50 - 15° W). Here the vertical distributions of light and nutrients control the abundance and growth of autotrophic algae in the thermically stratified water column. This phytoplankton is numerically dominated by the prokaryotic picoplankters Synechococcus spp. and Prochlorococcus spp., which are smaller than 2 ?m. The productivity of the 100 to 150 m deep euphotic zone can be high, because a high heterotrophic/autotrophic biomass ratio induces a rapid regeneration of nutrients and inorganic carbon. Primary grazers are mainly micro-organisms such as heterotrophic nannoflagellates and ciliates, which feed on the small algae and on bacteria. Heterotrophic bacteria can outnumber the autotrophic algae, because their number is related to the substrate pools of dissolved and particulate dead organic matter. These DOC and detritus pools reach equilibrium at a concentration, where the rate of their production (proportional to algal biomass) equals their mineralisation and sinking rate (proportional to the concentration and weight of POC and detritus). At a relatively low value of the weight-specific loss rates, the equilibrium concentration of these carbon pools and their load of bacteria can be high. The bacterial productivity is proportional to the mineralisation rate, which in a steady state can never be higher than the rate of primary production. Hence the ratio in turnover rate of bacteria and autotrophs tends to be reciprocally proportional to their biomass ratio.

Geochemical analysis of sediments and interstitial water in sediment cores from the North-West African continental margin and from the Central Pacific

A. Continental slope sediments off Spanish-Sahara and Senegal contain up to 4% organic carbon and up to 0.4% total nitrogen. The highest concentrations were found in sediments from water depths between 1000 and 2000 m. The regional and vertical distribution of organic matter differs significantly. Off Spanish-Sahara the organic matter content of sediment deposited during glacial times (Wuerm, Late Riss) is high whereas sediments deposited during interglacial times (Recent, Eem) are low in organic matter. Opposite distribution was found in sediments off Senegal. The sediments contain 30 to 130 ppm of fixed nitrogen. In most sediments this corresponds to 2-8 % of the total nitrogen. Only in sediments deposited during interglacial times off Spanish-Sahara up to 20 % of the total nitrogen is contained as inorganically bound nitrogen. Positive correlations of the fixed nitrogen concentrations to the amounts of clay, alumina, and potassium suggest that it is primarily fixed to illites. The amino acid nitrogen and hexosamine nitrogen account for 17 to 26 % and 1.3 to 2.4 %, respectively of the total nitrogen content of the sediments. The concentrations vary between 200 and 850 ppm amino acid nitrogen and 20 to 70 ppm hexosamine nitrogen, both parallel the fluctiations of organic matter in the sediment. Fulvic acids, humic acids, and the total organic matter of the sediments may be clearly differentiated from one another and their amino acid and hexosamine contents and their amino acid composition: a) Fulvic acids contain only half as much amino acids as humic acids b) The molar amino acid/hexosamine ratios of the fulvic acids are half those of the humic acids and the total organic matter of the sediment c) The amino acid spectra of fulvic acids are characterized by an enrichment of aspartic acid, alanine, and methionine sulfoxide and a depletion of glycine, valine, isoleucine, leucine, tyrosine, phenylalanine, lysine, and arginine compared to the spectra of the humic acids and those of the total organic matter fraction of the sediment. d) The amino acid spectra of the humic acids and those of the total organic matter fraction of the sediments are about the same with the exception that arginine is clearly enriched in the total organic matter. In general, as indicated by the amino compounds humic acids resemble closer the total organic matter composition than the low molecular fulvic acids do. This supports the general idea that during the course of diagenesis in reducing sediments organic matter stabilizes from a fulvic-like structure to humic-like structure and finally to kerogen. The decomposition rates of single aminio acids differ significantly from one another. Generally amino acids which are preferentially contained in humic acids and the total organic matter fraction show a smaller loss with time than those preferably well documented in case of the basic amino acids lysine and arginine which- although thermally unstable- are the most stable amino acids in the sediments. A favoured incorporation of these compounds into high molecular substances as well as into clay minerals may explain their relatively high "stability" in the sediment. The nitrogen loss from the sediments due to the activity of sulphate-reducing bacteria amounts to 20-40 % of the total organic nitrogen now present. At least 40 % of the organic nitrogen which is liberated by sulphate-reducing bacteria can be explained ny decomposition of amino acids alone. B. Deep-sea sediments from the Central Pacific The deep-seas sediments contain 1 to 2 orders of magnitude less organic matter than the continental slope sediments off NW Africa, i.e. 0.04 to 0.3 % organic carbon. The fixed nitrogen content of the deep-sea sediments ranges from 60 to 270 ppm or from 20 to 45 % of the total nitrogen content. While ammonia is the prevailing inorganic nitrogen compound in anoxic pore waters, nitrate predominates in the oxic environment of the deep-sea sediments. Near the sediment/water interface interstital nitrate concentrations of around 30 µg-at. N/l were recorded. These generally increase with sediment depth by 10 to 15 µg-at. NO3- N/l. This suggests the presence of free oxygen and the activity of nitrifying bacteria in the interstitial waters. The ammonia content of the interstitial water of the oxic deep-sea sediments ranges from 2 to 60 µg-at. N/l and thus is several orders of magnitude less than in anoxic sediments. In contrast to recorded nitrate gradients towards the sediments/water interface, there are no ammonia concentration gradients. However, ammonia concentrations appear to be characteristic for certain regional areas. It is suggested that this regional differentiation is caused by ion exchange reactions involving potassium and ammonium ions rather than by different decomposition rates of organic matter. C. C/N ratios All estimated C/N ratios of surface sediments vary between 3 and 9 in the deep-sea and the continental margin, respectively. Whereas the C/N ratios generally increase with depth in the sediment cores off NW Africa they decrease in the deep-sea cores. The lowest values of around 1.3 were found in the deeper sections of the deep-sea cores, the highest of around 10 in the sediments off NW Africa. The wide range of the C/N ratios as well as their opposite behaviour with increasing sediment depth in both the deep-sea and continental margin sediment cores, can be attributed mainly to the combination of the following three factors: 1. Inorganic and organic substances bound within the latticed of clay minerals tend to decrease the C/N ratios. 2. Organic matter not protected by absorption on the clay minerals tends to increase C/N ratios 3. Diagenetic alteration of organic matter by micro-organisms tends to increase C/N ratios through preferential loss of nitrogen The diagenetic changes of the microbially decomposable organic matter results in both oxic and anoxic environments in a preferential loss of nitrogen and hence in higher C/N ratios of the organic fraction. This holds true for most of the continental margin sediments off NW Africa which contain relatively high amounts of organic matter so that factors 2 and 3 predominate there. The relative low C/N ratios of the sediments deposited during interglacial times off Spanish-Sahara, which are low in organic carbon, show the increasing influence of factor 1 - the nitrogen-rich organic substances bound to clay minerals. In the deep-sea sediments from the Central Pacific this factor completely predominates so that the C/N rations of the sediments approach that of the substance absorbed to clay minerals with decreasing organic matter content. In the deeper core sections the unprotected organic matter has been completely destroyed so that the C/N ratios of the total sediments eventually fall into the same range as those of the pure clay mineral fraction.