Analysis of Polyhydroxyalkanoates Granules in Haloferax mediterranei by Double-Fluorescence Staining with Nile Red and SYBR Green by Confocal Fluorescence Microscopy.

Haloferaxmediterranei is a haloarchaeon of high interest in biotechnology because it produces and mobilizes intracellular polyhydroxyalkanoate (PHA) granules during growth under stress conditions (limitation of phosphorous in the culture media), among other interesting metabolites (enzymes, carotenoids, etc.). The capability of PHA production by microbes can be monitored with the use of staining-based methods. However, the staining of haloarchaea cells is a challenging task; firstly, due to the high ionic strength of the medium, which is inappropriate for most of dyes, and secondly, due to the low permeability of the haloarchaea S-layer to macromolecules. In this work, Haloferax mediterranei is used as a halophilic archaeon model to describe an optimized protocol for the visualization and analysis of intracellular PHA granules in living cells. The method is based on double-fluorescence staining using Nile red and SYBR Green by confocal fluorescence microscopy. Thanks to this method, the capability of PHA production by new haloarchaea isolates could be easily monitored.

Synthesis and Characterization of New Layered Double Hydroxide-Polyolefin Film Nanocomposites with Special Optical Properties.

In this study, we have synthesized new double layered hydroxides to be incorporated to low density polyethylene thermoplastic matrix. These new composites present promising applications as materials to build greenhouses due to the enhancement of their optical properties. A characterization of the modified nanoclay has been performed by means of X-ray fluorescence (XRF), X-ray Diffraction (XRD), Thermogravimetric analysis (TGA), and Fourier-transform infrared spectroscopy (FTIR). We have prepared a series of polyolefin-based films to evaluate the effect of the addition of a whitening agent (disodium 2,2'-((1,1'-biphenyl)-4,4'-diyldivinylene)bis(benzenesulfonate)), the modified hydrotalcite-like material and a commercial dispersant. The rheological and mechanical characterization of the films have proved that the inclusion of the modified-layered double hydroxides (LDHs) do not substantially affect the processing and mechanical performance of the material. On the other hand, optical properties of the nanocomposites are improved by reducing the transmission in the UVA region.