Chile 50 años: Inolvidando

Chile 50 years: Unforgettin

Direct analysis of microbial populations in soil and freshwater aquifers using nucleic acid based techniques

The first DNA-based methods for direct quantification of soil protozoa, and a DNA-based quantification method to describe the spread of phenanthrene-degrading bacteria in soil and freshwater aquifers, have recently been developed at the BIOPRO Research Centre at the Geological Survey of Denmark and Greenland (GEUS). Well-known genes for phenoxyalcanoic acid degradation have been used to monitor the in situ degradation of phenoxyalcanoic acid pesticides. Studies have been initiated on the short-lived mRNA molecules that are expected to provide a shortcut to the understanding of low, yet important, microbial activity in geological samples. This article reviews recent developments in techniques based on analysis of nucleic acids from soils and aquifers. Analytical work has been carried out mainly on soil samples from a former asphalt production plant at Ringe (Fig. 1). The Ringe plant constitutes one of the most polluted industrial sites in Denmark, and is a priority site of studies by the BIOPRO Research Centre. Although rich in carbon, the Ringe subsoil is an oligotrophic environment due to the high content of polycyclic aromatic hydrocarbons (PAH). This is an environment where the supply of nutrients to microorganisms is low, leading to slow growth, low total numbers of microorganisms and small cells. To study microbial communities of oligotrophic environments, analytical methods with low detection limits are needed. Until recently, microorganisms of natural environments were mainly studied by cultivation-dependent methods. However, microorganisms that can be cultured on agar plates are now known to represent only a small fraction of the total microbial community. Modern methods, therefore, need to be based on the detection of biomolecules in the microorganisms rather than being dependent on growth of the microorganisms. The best available techniques are based on DNA and RNA molecules (Fig. 2), which due to their high level of resolution allow closely related organisms or functional genes to be distinguished. In the following review, examples are given of applications of these nucleic acid based methods

Comparison of DNA extraction kits for PCR-DGGE analysis of human intestinal microbial communities from fecal specimens

<p>Abstract</p> <p>Background</p> <p>The influence of diet on intestinal microflora has been investigated mainly using conventional microbiological approaches. Although these studies have advanced knowledge on human intestinal microflora, it is imperative that new methods are applied to facilitate scientific progress. Culture-independent molecular fingerprinting method of Polymerase Chain Reaction and Denaturing Gradient Gel Electrophoresis (PCR-DGGE) has been used to study microbial communities in a variety of environmental samples. However, these protocols must be optimized prior to their application in order to enhance the quality and accuracy of downstream analyses. In this study, the relative efficacy of four commercial DNA extraction kits (Mobio Ultra Clean^®Fecal DNA Isolation Kit, M; QIAamp^®DNA Stool Mini Kit, Q; FastDNA^®SPIN Kit, FSp; FastDNA^®SPIN Kit for Soil, FSo) were evaluated. Further, PCR-DGGE technique was also assessed for its feasibility in detecting differences in human intestinal bacterial fingerprint profiles.</p> <p>Method</p> <p>Total DNA was extracted from varying weights of human fecal specimens using four different kits, followed by PCR amplification of bacterial 16S rRNA genes, and DGGE separation of the amplicons.</p> <p>Results</p> <p>Regardless of kit, maximum DNA yield was obtained using 10 to 50 mg (wet wt) of fecal specimens and similar DGGE profiles were obtained. However, kits FSp and FSo extracted significantly larger amounts of DNA per g dry fecal specimens and produced more bands on their DGGE profiles than kits M and Q due to their use of bead-containing lysing matrix and vigorous shaking step. DGGE of 16S rRNA gene PCR products was suitable for capturing the profiles of human intestinal microbial community and enabled rapid comparative assessment of inter- and intra-subject differences.</p> <p>Conclusion</p> <p>We conclude that extraction kits that incorporated bead-containing lysing matrix and vigorous shaking produced high quality DNA from human fecal specimens (10 to 50 mg, wet wt) that can be resolved as bacterial community fingerprints using PCR-DGGE technique. Subsequently, PCR-DGGE technique can be applied for studying variations in human intestinal microbial communities.</p

Chile: memoria de las voces

Gracias a la vida que me ha dado tantome dio el corazón que agita su marcocuando miro el fruto del cerebro humano,cuando miro el bueno tan lejos del malo,cuando miro el fondo de tus ojos claros. Violeta Parra, campesina de Parral, sur de Chile, nos enseñó las raíces del canto chileno y latinoamericano. Partió resoluta, sin dinero y una grabadora por territorios imposibles, buscando gente sencilla, recopilando el tesoro oral que se transmitía en el campo de una generación a otra: canciones, ri..

Crónica de un Taller muy particular

“Todos los sábados llueve torrencialmente en Milán y aún más en el Barrio Chino“. Eso pienso mis primeros dos días en Italia, después de que Laura Scarabelli y su pareja me buscaran en el aeropuerto, me mostraran amablemente el barrio y me llevaran a mi departamentito para mis próximas tres semanas. Nos despedimos hasta el lunes 29 de febrero, emblemática fecha y comienzo de mi taller