Miniaturized bacterial biosensor system for arsenic detection holds great promise for making integrated measurement device.

Combining bacterial bioreporters with microfluidics systems holds great promise for in-field detection of chemical or toxicity targets. Recently we showed how Escherichia coli cells engineered to produce a variant of green fluorescent protein after contact to arsenite and arsenate can be encapsulated in agarose beads and incorporated into a microfluidic chip to create a device for in-field detection of arsenic, a contaminant of well known toxicity and carcinogenicity in potable water both in industrialized and developing countries. Cell-beads stored in the microfluidics chip at -20°C retained inducibility up to one month and we were able to reproducibly discriminate concentrations of 10 and 50 μg arsenite per L (the drinking water standards for European countries and the United States, and for the developing countries, respectively) from the blank in less than 200 minutes. We discuss here the reasons for decreasing bioreporter signal development upon increased storage of cell beads but also show how this decrease can be reduced, leading to a faster detection and a longer lifetime of the device.

Generalist hydrocarbon-degrading bacterial communities in the oil-polluted water column of the North Sea.

The aim of this work was to determine the effect of light crude oil on bacterial communities during an experimental oil spill in the North Sea and in mesocosms (simulating a heavy, enclosed oil spill), and to isolate and characterize hydrocarbon-degrading bacteria from the water column. No oil-induced changes in bacterial community (3 m below the sea surface) were observed 32 h after the experimental spill at sea. In contrast, there was a decrease in the dominant SAR11 phylotype and an increase in Pseudoalteromonas spp. in the oiled mesocosms (investigated by 16S rRNA gene analysis using denaturing gradient gel electrophoresis), as a consequence of the longer incubation, closer proximity of the samples to oil, and the lack of replenishment with seawater. A total of 216 strains were isolated from hydrocarbon enrichment cultures, predominantly belonging to the genus Pseudoaltero monas; most strains grew on PAHs, branched and straight-chain alkanes, as well as many other carbon sources. No obligate hydrocarbonoclastic bacteria were isolated or detected, highlighting the potential importance of cosmopolitan marine generalists like Pseudoalteromonas spp. in degrading hydrocarbons in the water column beneath an oil slick, and revealing the susceptibility to oil pollution of SAR11, the most abundant bacterial clade in the surface ocean.

Oxidative stress in gastric mucosa of asymptomatic humans infected with Helicobacter pylori: effect of bacterial eradication.

BACKGROUND: Inducible nitric oxide synthase (iNOS) and interleukin 8 (IL-8) are positive in approximately 50% of Helicobacter pylori-related diseases but it is not clear whether oxidative stress is also present in H. pylori asymptomatic humans. Our aim was to study the expression of iNOS, superoxide dismutase, catalase and IL-8 production in H. pylori-infected asymptomatic humans, and to investigate the effect of eradication of H. pylori. MATERIALS AND METHODS: Biopsies of corpus and antrum of asymptomatic H. pylori positive and negative humans served for determination of the gastritis score and H. pylori status; iNOS was measured by reverse transcriptase polymerase chain reaction and immunohistochemistry and superoxide dismutase and catalase by immunohistochemistry. IL-8 in biopsies was assessed by enzyme-linked immunosorbent assay. RESULTS: Immunostaining of iNOS, catalase and superoxide dismutase was significantly associated with H. pylori infection and was localized to inflammatory cells. IL-8 concentrations were greater in the H. pylori positive than H. pylori negative group and decreased after bacterial eradication. A decrease in staining for iNOS and catalase was observed after H. pylori eradication. CONCLUSIONS: INOS and antioxidant enzymes are present in gastric biopsies of asymptomatic H. pylori positive humans. Eradication caused a significant decrease in staining for iNOS and catalase. These results indicate that oxidative stress occurs in asymptomatic patients and can be modulated by H. pylori eradication.

The dynamics of biofilm bacterial communities is driven by flow wax and wane in a temporary stream

Biofilm communities are exposed to long periods of desiccation in temporary streams. We investigated how water flow intermittency affected the bacterial community structure colonizing three different streambed compartments in a Mediterranean stream. Massive parallel sequencing revealed different bacterial communities in biofilms from sand sediments and cobbles. Bacterial communities were similar (62% of shared operational taxonomic units) in the epipsammic and hyporheic biofilms, and more diverse than those in the epilithic biofilms. The non-flow phase caused a decrease of bacterial diversity in the biofilms, when communities included only bacterial taxa assumed to be adapted to water stress. The most sensitive bacterial communities to flow intermittency were in the epilithic, where the exposure to physical stress was the highest. In sand sediments a wide group of bacterial taxa was tolerant to desiccation. During non-flow the proliferation of opportunistic taxa in the superficial compartments evidenced the biological link with the terrestrial environment. Bacterial communities better tolerate rewetting than desiccation, since a major number of taxa tolerant to rewetting occurred in all biofilms. Overall, bacterial communities in sandy compartments showed higher resistance to flow intermittency than those in epilithic biofilms

Étude des propriétés de transport dans les hydrogels de curdlan

Les hydrogels de polysaccharide sont des biomatériaux utilisés comme matrices à libération contrôlée de médicaments et comme structures modèles pour l’étude de nombreux systèmes biologiques dont les biofilms bactériens et les mucus. Dans tous les cas, le transport de médicaments ou de nutriments à l’intérieur d’une matrice d’hydrogel joue un rôle de premier plan. Ainsi, l’étude des propriétés de transport dans les hydrogels s’avère un enjeu très important au niveau de plusieurs applications. Dans cet ouvrage, le curdlan, un polysaccharide neutre d’origine bactérienne et formé d’unités répétitives β-D-(1→3) glucose, est utilisé comme hydrogel modèle. Le curdlan a la propriété de former des thermogels de différentes conformations selon la température à laquelle une suspension aqueuse est incubée. La caractérisation in situ de la formation des hydrogels de curdlan thermoréversibles et thermo-irréversibles a tout d’abord été réalisée par spectroscopie infrarouge à transformée de Fourier (FT-IR) en mode réflexion totale atténuée à température variable. Les résultats ont permis d’optimiser les conditions de gélation, menant ainsi à la formation reproductible des hydrogels. Les caractérisations structurales des hydrogels hydratés, réalisées par imagerie FT-IR, par microscopie électronique à balayage en mode environnemental (eSEM) et par microscopie à force atomique (AFM), ont permis de visualiser les différentes morphologies susceptibles d’influencer la diffusion d’analytes dans les gels. Nos résultats montrent que les deux types d’hydrogels de curdlan ont des architectures distinctes à l’échelle microscopique. La combinaison de la spectroscopie de résonance magnétique nucléaire (RMN) à gradients pulsés et de l’imagerie RMN a permis d’étudier l’autodiffusion et la diffusion mutuelle sur un même système dans des conditions expérimentales similaires. Nous avons observé que la diffusion des molécules dans les gels est ralentie par rapport à celle mesurée en solution aqueuse. Les mesures d’autodiffusion, effectuées sur une série d’analytes de diverses tailles dans les deux types d’hydrogels de curdlan, montrent que le coefficient d’autodiffusion relatif décroit en fonction de la taille de l’analyte. De plus, nos résultats suggèrent que l’équivalence entre les coefficients d’autodiffusion et de diffusion mutuelle dans les hydrogels de curdlan thermo-irréversibles est principalement due au fait que l’environnement sondé par les analytes durant une expérience d’autodiffusion est représentatif de celui exploré durant une expérience de diffusion mutuelle. Dans de telles conditions, nos résultats montrent que la RMN à gradients pulsés peut s’avérer une approche très avantageuse afin de caractériser des systèmes à libération contrôlée de médicaments. D’autres expériences de diffusion mutuelle, menées sur une macromolécule de dextran, montrent un coefficient de diffusion mutuelle inférieur au coefficient d’autodiffusion sur un même gel de curdlan. L’écart mesuré entre les deux modes de transport est attribué au volume différent de l’environnement sondé durant les deux mesures. Les coefficients d’autodiffusion et de diffusion mutuelle similaires, mesurés dans les deux types de gels de curdlan pour les différents analytes étudiés, suggèrent une influence limitée de l’architecture microscopique de ces gels sur leurs propriétés de transport. Il est conclu que les interactions affectant la diffusion des analytes étudiés dans les hydrogels de curdlan se situent à l’échelle moléculaire.

Étude de l'influence de la réassociation en surface des atomes N et O sur l'inactivation des spores bactériennes dans une post-décharge N2-O2 basse pression en flux

Le recours au plasma pour stériliser des dispositifs médicaux (DM) est un domaine de recherche ne datant véritablement que de la fin des années 1990. Les plasmas permettent, dans les conditions adéquates, de réaliser la stérilisation à basse température (≤ 65°C), tel qu’exigé par la présence de polymères dans les DM et ce contrairement aux procédés par chaleur, et aussi de façon non toxique, contrairement aux procédés chimiques comme, par exemple, l’oxyde d’éthylène (OEt). Les laboratoires du Groupe de physique des plasmas à l’Université de Montréal travaillent à l’élaboration d’un stérilisateur consistant plus particulièrement à employer les effluents d’une décharge N2-%O2 basse pression (2-8 Torrs) en flux, formant ce que l’on appelle une post-décharge en flux. Ce sont les atomes N et O de cette décharge qui viendront, dans les conditions appropriées, entrer en collisions dans la chambre de stérilisation pour y créer des molécules excitées NO*, engendrant ainsi l’émission d’une quantité appréciable de photons UV. Ceux-ci constituent, dans le cas présent, l’agent biocide qui va s’attaquer directement au bagage génétique du micro-organisme (bactéries, virus) que l’on souhaite inactiver. L’utilisation d’une lointaine post-décharge évite du même coup la présence des agents érosifs de la décharge, comme les ions et les métastables. L’un des problèmes de cette méthode de stérilisation est la réduction du nombre de molécules NO* créées par suite de la perte des atomes N et O, qui sont des radicaux connus pour interagir avec les surfaces, sur les parois des matériaux des DM que l’on souhaite stériliser. L’objectif principal de notre travail est de déterminer l’influence d’une telle perte en surface, dite aussi réassociation en surface, par l’introduction de matériaux comme le Téflon, l’acier inoxydable, l’aluminium et le cuivre sur le taux d’inactivation des spores bactériennes. Nous nous attendons à ce que la réassociation en surface de ces atomes occasionne ainsi une diminution de l’intensité UV et subséquemment, une réduction du taux d’inactivation. Par spectroscopie optique d’émission (SOE), nous avons déterminé les concentrations perdues de N et de O par la présence des matériaux dans le stérilisateur, ainsi que la diminution de l’émission UV en découlant. Nous avons observé que cette diminution des concentrations atomiques est d’autant plus importante que les surfaces sont catalytiques. Au cours de l’étude du phénomène de pertes sur les parois pour un mélange N2-%O2 nous avons constaté l’existence d’une compétition en surface entre les atomes N et O, dans laquelle les atomes d’oxygènes semblent dominer largement. Cela implique qu’au-delà d’un certain %O2 ajouté à la décharge N2, seuls les atomes O se réassocient en surface. Par ailleurs, l’analyse des courbes de survie bi-phasiques des micro-organismes a permis d’établir une étroite corrélation, par lien de cause à effet, entre la consommation des atomes N et O en surface et la diminution du taux d’inactivation des spores dans la première phase. En revanche, nous avons constaté que notre principal agent biocide (le rayonnement ultraviolet) est moins efficace dans la deuxième phase et, par conséquent, il n’a pas été possible d’établir un lien entre la diminution des concentrations et le taux d’inactivation de cette phase-là.

Mass production of nitrifying bacterial consortia for the rapid establishment of nitrification in saline recirculating aquaculture systems

Two distinct nitrifying bacterial consortia, namely an ammonia oxidizing non-penaeid culture (AMO NPCU-1) and an ammonia oxidizing penaeid culture (AMOPCU-1), have been mass produced in a nitrifying bacterial consortia production unit (NBCPU). The consortia, maintained at 4 C were activated and cultured in a 2 l fermentor initially. At this stage the net biomass (0.105 and 0.112 g/l), maximum specific growth rate (0.112 and 0.105/h) and yield coefficients (1.315 and 2.08) were calculated respectively, for AMONPCU-1 and AMOPCU-1 on attaining stationary growth phase. Subsequently on mass production in a 200 l NBCPU under optimized culture conditions, the total amounts of NH4 ?–N removed by AMONPCU-1 and AMOPCU-1 were 1.948 and 1.242 g/l within 160 and 270 days, respectively. Total alkalinity reduction of 11.7–14.4 and 7.5–9.1 g/l were observed which led to the consumption of 78 and 62 g Na2CO3. The yield coefficient and biomass of AMONPCU-1 were 0.67 and 125.3 g/l and those of AMOPCU-1 were 1.23 and 165 g/l. The higher yield coefficient and growth rate of AMOPCU-1 suggest better energy conversion efficiency and higher CO2 fixation potential. Both of the consortia were dominated by Nitrosomonas-like organisms. The consortia may find application in the establishment of nitrification within marine and brackish water culture systems.

Development of nitrifying bacterial consortia for immobilizing in nitrifying bioreactors designed for penaeid and non-penaeid larval rearing systems in the tropics

Two ammonia oxidizing (AMOPCU-1 and AMONPCU-1) and two nitrite oxidizing (NIOPCU-1 and NIONPCU-1) consortia for activating nitrifying bioreactors and thereby establishing nitrification in penaeid and non-penaeid hatchery systems were developed by enrichment. For further amplification of the consortia a simple medium having seawater (either salinity 30 ‰ or 15 ‰) as base, supplemented with NH4+-N/NO2--N and PO4- and pH adjusted to 8 was identified. During the amplification in a fermentor the consortia exhibited excessive wall growth and diminished their yield coefficient posing difficulty in harvesting the cells completely. The consortia consisted of both Gram negative and Gram-positive bacterial cells embedded in a mucilaginous matrix of glycocalyx - like material presumably composed of polysaccharides. The consortia besides being useful in activating nitrifying bioreactors developed for shrimp/prawn hatchery systems can also be used as bioaugmentors in the bioremediation of ammonia and nitrite toxicity in aquaculture systems.

Factors affecting biological control effectiveness of Pasteuria penetrans in Meloidogyne javanica and the bacterial development in the nematode body

The invasion and infectivity of Meloidogyne javanica juveniles (J2) encumbered with spore of Pasteuria Penetrans were influenced by the temperature and the time J2 were in the soil before exposure to roots. The percentage of infected females decreased as the time juveniles spent in soil increased. When spore encumbered J2 were maintained at 30 degrees C the decrease in infection was greater than that at 18 degrees C. The thermal time requirements and the base temperature for P. penetrans development were estimated. The rate of development followed an exponential curve between 21 and 36 degrees C and the base temperature for development was estimated by extrapolation to be 18.5 degrees C. The effect of integrating a nematode resistant tomato cultivar with the biocontrol agent P. penetrans also was investigated. The ability of the biocontrol agent to reduce numbers of root-knot nematodes was dependent on the densities of the nematode and P. penetrans spores in the soil.

The pst operon of enteropathogenic Escherichia coli enhances bacterial adherence to epithelial cells

Enteropathogenic Escherichia coli (EPEC) adheres in vivo and in vitro to epithelial cells. Two main adhesins, the bundle-forming pilus and intimin, encoded by the Up operon and eae, respectively, are responsible for the localized and the intimate adherence phenotypes. Deletion of the pst operon of EPEC abolishes the transport of inorganic phosphate through the phosphate-specific transport system and causes the constitutive expression of the PHO regulon genes. In the absence of pst there is a decrease in the expression of the main EPEC adhesins and a reduction in bacterial adherence to epithelial cells in vitro. This effect is not related to PHO constitutivity, because a Delta pst phoB double mutant that is defective in the transcription of the PHO genes also displayed low levels of adherence and expression of adhesins. Likewise, a PHO-constitutive phoR mutation did not affect bacterial adherence. The expression of the per operon, which encodes the Up and ler regulators PerA and PerC, is also negatively affected by the pst deletion. Overall, the data presented here demonstrate that the pst operon of EPEC plays a positive role in the bacterial adherence mechanism by increasing the expression of perA and perC and consequently the transcription of bfp and eae.

New bacterial cellulose nanocomposites prepared by in situ radical polymerization of methacrylate monomers

Bacterial cellulose/polymethacrylate nanocomposites have received attention in numerous areas of study and in a variety of applications. The attractive properties of methacrylate polymers and bacterial cellulose, BC, allow the synthesis of new nanocomposites with distinct characteristics. In this study, BC/poly(glycidylmethacrylate) (BC/PGMA) and BC/poly(ethyleneglycol)methacrylate (BC/PPEGMA) nanocomposites were prepared through in situ free radical polymerization of GMA and PEGMA, respectively. Ammonium persulphate (APS) was used as an initiator and N,N’methylenebisacrilamide (MBA) was used as a crosslinker in BC/PGMA. Chemical composition, morphology, thermal stability, water absorption, mechanic and surface properties were determined through specific characterization techniques. The optimal polymerization was obtained at (1:2) for BC/PGMA, (1:2:0.2) ratio for BC/GMA/MBA and (1:20) for BC/PPEGMA, with 0.5% of initiator at 60 ºC during 6 h. A maximum of 67% and 87% of incorporation percentage was obtained, respectively, for the nanocomposites BC/PGMA/MBA and BC/PPEGMA. BC/PGMA nanocomposites exhibited an increase of roughness and compactation of the three-dimensional structure, an improvement in the thermal and mechanical properties, and a decrease in their swelling ability and crystallinity. On the other hand, BC/PPEGMA showed a decrease of stiffness of three-dimensional structure, improvement in thermal and mechanical properties, an increase in their swelling ability and a decrease the crystallinity. Both BC/polymethacrylate nanocomposites exhibited a basic surface character. The acid treatment showed to be a suitable strategy to modifiy BC/PGMA nanocomposites through epoxide ring-opening reaction mechanism. Nanocomposites became more compact, smooth and with more water retention ability. A decrease in the thermal and mechanical proprieties was observed. The new nanocomposites acquired properties useful to biomedical applications or/and removal of heavy metals due to the presence of functional groups.

Synthesis and characterization of microcrystalline cellulose produced from bacterial cellulose

In this study, microcrystalline cellulose (MCC) was prepared from the acid hydrolysis of bacterial cellulose (BC) produced in culture medium of static Acetobacter xylinum. The MCC-BC produced an average particle size between 70 and 90 mu m and a degree of polymerization (DP) of 250. The characterization of samples was performed by thermogravimetric analysis, X-ray diffraction, and scanning electron microscopy (SEM). The MCC shows a lower thermal stability than the pristine cellulose, which was expected due to the decrease in the DP during the hydrolysis process. In addition, from X-ray diffractograms, we observed a change in the crystalline structure. The images of SEM for the BC and MCC show clear differences with modifications of BC fiber structure and production of particles with characteristics similar to commercial MCC.

The effects of bacterial leaching on metal partitioning in sewage sludge

The partitioning of Mn, Al, Zn, Cu and Ti ions in municipal sewage sludge was investigated before and after bioleaching processes effectuated by Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans. Oxidation reduction potential increase and pH decrease were obtained as a result of bacterial activity. A less pronounced and constant decrease was obtained with A. ferrooxidans, whereas A. thiooxidans presented a lag phase before a steep pH decrease. Metal solubilization was accomplished in experimental systems supplemented with energy source, Fe2+ for A. ferrooxidans and S-0 for A. thiooxidans. Solubilization efficiency differed for each metal except for Al, and was relatively similar for either organism. Metal partitioning was conducted using five-step sequential extraction procedure before and after the bioleaching. The results indicated that Zn and Mn ions were mostly associated with the organic fraction, whereas Cu, Al and Ti ions with the sulphide/ residue fraction. The bioleaching process caused prompt solubilization of metals mostly associated with the more labile fractions (exchangeable, adsorbed and organically bound metals), whereas those associated to the less labile ones (EDTA and sulphide/residue fractions) were exchanged towards more labile fractions.

Diffusion coefficient of K+ in ion exchanged glasses calculated from the refractive index and the Vickers hardness profiles

The top faces of float glass samples were exposed to vapors resulting from the decomposition of KNO3 at 565 degrees C for up to 32 h. X-ray dispersive spectra (EDS) show that K+ ions migrate into the glass. The K+ concentration profile was obtained and its diffusion coefficient was calculated by the Boltzmann-Matano technique. The mean diffusion coefficient was approximately 10 X 10(-11) cm(2) s(-1). It was observed that the refractive index and the Vickers hardness decrease with the depth (after the removal of successive layers), and their profiles were thus obtained. These profiles enabled the calculation of the diffusion coefficient of K+ through the Boltzmann-Matano technique, with mean results ranging between 6 x 10(-11) and 30 x 10(-11) cm(2) s(-1). (c) 2006 Elsevier B.V. All rights reserved.

Transparent bacterial cellulose-boehmite-epoxi-siloxane nanocomposites

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