Assessment of microbial community structure changes by amplified ribosomal DNA restriction analysis (ARDRA)

Amplified ribosomal DNA restriction analysis (ARDRA) is a simple method based on restriction endonuclease digestion of the amplified bacterial 16S rDNA. In this study we have evaluated the suitability of this method to detect differences in activated sludge bacterial communities fed on domestic or industrial wastewater, and subject to different operational conditions. The ability of ARDRA to detect these differences has been tested in modified Ludzack-Ettinger (MLE) configurations. Samples from three activated sludge wastewater treatment plants (WWTPs) with the MLE configuration were collected for both oxic and anoxic reactors, and ARDRA patterns using double enzyme digestions AluI+MspI were obtained. A matrix of Dice similarity coefficients was calculated and used to compare these restriction patterns. Differences in the community structure due to influent characteristics and temperature could be observed, but not between the oxic and anoxic reactors of each of the three MLE configurations. Other possible applications of ARDRA for detecting and monitoring changes in activated sludge systems are also discussed

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry, a revolution in clinical microbial identification.

Until recently, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) techniques for the identification of microorganisms remained confined to research laboratories. In the last 2 years, the availability of relatively simple to use MALDI-TOF MS devices, which can be utilized in clinical microbiology laboratories, has changed the laboratory workflows for the identification of pathogens. Recently, the first prospective studies regarding the performance in routine bacterial identification showed that MALDI-TOF MS is a fast, reliable and cost-effective technique that has the potential to replace and/or complement conventional phenotypic identification for most bacterial strains isolated in clinical microbiology laboratories. For routine bacterial isolates, correct identification by MALDI-TOF MS at the species level was obtained in 84.1-93.6% of instances. In one of these studies, a protein extraction step clearly improved the overall valid identification yield, from 70.3% to 93.2%. This review focuses on the current state of use of MALDI-TOF MS for the identification of routine bacterial isolates and on the main difficulties that may lead to erroneous or doubtful identifications.

Biomassa microbiana e produção de C-CO2 e N mineral de um podzólico vermelho-escuro submetido a diferentes sistemas de manejo: Microbial biomass and C-CO2 and mineral nitrogen production in paleudult soil cultivated under different management systems

Os sistemas de manejo, com diferenças no revolvimento do solo e na composição dos resíduos vegetais, alteram as propriedades biológicas do solo, com reflexos na qualidade do solo e na produtividade das culturas. Com vistas em medir estas alterações nas propriedades biológicas do solo, a biomassa e a atividade microbiana foram avaliadas em um Podzólico Vermelho-Escuro, em Eldorado do Sul (RS), utilizando diferentes preparos (convencional, reduzido e plantio direto) e dois sistemas de sucessões de culturas (aveia preta + vica/milho + caupi e aveia/milho). As avaliações foram realizadas em quatro épocas, durante 12 meses, e em duas profundidades (0-5 e 5-15 cm). O carbono da biomassa microbiana foi analisado pelo método de fumigação-incubação, e a atividade microbiana, pela produção de C-CO2 e N mineral, após 60 dias de incubação. As diferenças na biomassa e na atividade microbiana, entre os sistemas de manejo, foram mais pronunciadas na camada de 0-5 cm. Nesta camada de solo, observaram-se os maiores valores de biomassa e de atividade nos preparos conservacionistas e no sistema aveia + vica/milho + caupi. Dentre as variáveis estudadas, a mineralização de N mostrou-se a mais sensível aos manejos, à profundidade e à época de amostragem.

Surface topography and mixed-pixel effects on the simulated L-band brightness temperatures.

The impact of topography and mixed pixels on L-band radiometric observations over land needs to be quantified to improve the accuracy of soil moisture retrievals. For this purpose, a series of simulations has been performed with an improved version of the soil moisture and ocean salinity (SMOS) end-to-end performance simulator (SEPS). The brightness temperature generator of SEPS has been modified to include a 100-m-resolution land cover map and a 30-m-resolution digital elevation map of Catalonia (northeast of Spain). This high-resolution generator allows the assessment of the errors in soil moisture retrieval algorithms due to limited spatial resolution and provides a basis for the development of pixel disaggregation techniques. Variation of the local incidence angle, shadowing, and atmospheric effects (up- and downwelling radiation) due to surface topography has been analyzed. Results are compared to brightness temperatures that are computed under the assumption of an ellipsoidal Earth.

Europe and the Petersberg Tasks – Nation-State behavior through the prism of Strategic Culture

Les missions Petersberg són l'operatiu militar més ambiciós organitzat per la Unió Europea en el desenvolupament de la CSDP, Política Europea de Seguretat i Defensa. Amb l'objectiu d'aconseguir una organització efectiva y funcional d'aquestes missions, és desitjable que les cultures estratègiques dels diferents Estats membres siguin, en gran mesura, compatibles en benefici d'una cultura estratègica europea amb directrius clares. Aquest estudi compara les cultures estratègiques d'Alemanya, el Regne Unit i França en referència al seu nivell de compatibilitat contrastant-les amb dos casos recents, exemples paradigmàtics de cultures estratègiques integrals. D'aquesta manera, pretenem descriure les circumstàncies en què es desenvolupen les missions Petersberg.

Structure-function characterization and optimization of a plant-derived antibacterial peptide.

Crushed seeds of the Moringa oleifera tree have been used traditionally as natural flocculants to clarify drinking water. We previously showed that one of the seed peptides mediates both the sedimentation of suspended particles such as bacterial cells and a direct bactericidal activity, raising the possibility that the two activities might be related. In this study, the conformational modeling of the peptide was coupled to a functional analysis of synthetic derivatives. This indicated that partly overlapping structural determinants mediate the sedimentation and antibacterial activities. Sedimentation requires a positively charged, glutamine-rich portion of the peptide that aggregates bacterial cells. The bactericidal activity was localized to a sequence prone to form a helix-loop-helix structural motif. Amino acid substitution showed that the bactericidal activity requires hydrophobic proline residues within the protruding loop. Vital dye staining indicated that treatment with peptides containing this motif results in bacterial membrane damage. Assembly of multiple copies of this structural motif into a branched peptide enhanced antibacterial activity, since low concentrations effectively kill bacteria such as Pseudomonas aeruginosa and Streptococcus pyogenes without displaying a toxic effect on human red blood cells. This study thus identifies a synthetic peptide with potent antibacterial activity against specific human pathogens. It also suggests partly distinct molecular mechanisms for each activity. Sedimentation may result from coupled flocculation and coagulation effects, while the bactericidal activity would require bacterial membrane destabilization by a hydrophobic loop.

Application of real-time PCR for total airborne bacterial assessment : comparison with epifluorescence microscopy and culture-dependent methods

Traditional culture-dependent methods to quantify and identify airborne microorganisms are limited by factors such as short-duration sampling times and inability to count nonculturableor non-viable bacteria. Consequently, the quantitative assessment of bioaerosols is often underestimated. Use of the real-time quantitative polymerase chain reaction (Q-PCR) to quantify bacteria in environmental samples presents an alternative method, which should overcome this problem. The aim of this study was to evaluate the performance of a real-time Q-PCR assay as a simple and reliable way to quantify the airborne bacterial load within poultry houses and sewage treatment plants, in comparison with epifluorescencemicroscopy and culture-dependent methods. The estimates of bacterial load that we obtained from real-time PCR and epifluorescence methods, are comparable, however, our analysis of sewage treatment plants indicate these methods give values 270-290 fold greater than those obtained by the ''impaction on nutrient agar'' method. The culture-dependent method of air impaction on nutrient agar was also inadequate in poultry houses, as was the impinger-culture method, which gave a bacterial load estimate 32-fold lower than obtained by Q-PCR. Real-time quantitative PCR thus proves to be a reliable, discerning, and simple method that could be used to estimate airborne bacterial load in a broad variety of other environments expected to carry high numbers of airborne bacteria. [Authors]

Theme of "swinging" in shamanism and folk culture

V. E. Sharapov & D. A. Nesanelis

Prevention of vitamin K deficiency bleeding with three oral mixed micellar phylloquinone doses: results of a 6-year (2005-2011) surveillance in Switzerland.

In 2003, the Swiss guidelines to prevent vitamin K deficiency bleeding (VKDB) were adapted. As two oral doses (2 mg, hour/day 4) of mixed micellar VK preparation had failed to abolish late VKDB, a third dose (week 4) was introduced. This report summarizes the new guidelines acceptance by Swiss pediatricians and the results of a prospective 6-year surveillance to study their influence on the incidence of VKDB. The new guidelines acceptance by Swiss pediatricians was evaluated by a questionnaire sent to all pediatricians of the Swiss Society of Paediatrics. With the help of the Swiss Paediatric Surveillance Unit, the incidence of VKDB was monitored prospectively from July 1, 2005 until June 30, 2011. Over a 6-year period (458,184 live births), there was one case of early and four cases of late VKDB. Overall incidence was 1.09/10(5) (95 % confidence intervals (CI) 0.4-2.6). Late VKDB incidence was 0.87/10(5) (95 % CI 0.24-2.24). All four infants with late VKDB had an undiagnosed cholestasis at the time of bleeding; parents of 3/4 had refused VK prophylaxis, and in 1/4, the third VK dose had been forgotten. Compared with historical control who had received only two oral doses of mixed micellar VK (18 cases for 475,372 live births), the incidence of late VKDB was significantly lower with three oral doses (Chi(2),Yates correction, P = 0.007). CONCLUSION: VKDB prophylaxis with 3 × 2 mg oral doses of mixed micellar VK seems to prevent adequately infants from VKDB. The main risk factors for VKDB in breast-fed infants are parental VK prophylaxis refusal or an unknown cholestasis.