Colorimetric Detection of Metals Using CdS-NPs Synthesized by an Organic Extract of Aspergillus niger.

The use of cadmium sulfide nanoparticles (CdS-NPs) synthesized by fungi presents highly stable chemical and optical characteristics; this makes them a promising alternative for development of colorimetric methods for metal detection. Moreover, application of CdS-NPs is challenging due to the biological material used to carry out synthesis and coating is highly diverse; therefore, it is necessary to evaluate if such components are present in the biological material. Thus, the objective of this work was to detect metallic ions in synthetic water samples using CdS-NPs synthesized by the extract of Aspergillus niger. The conditions to produce fungal extracts were determined through a factorial design 23; additionally, biomolecules involved in metallic ions detection, synthesis, and coating of CdS-NPs were quantified; the studied biomolecules are NADH, sulfhydryl groups, proteins, and ferric reducing antioxidants (FRAP). CdS-NPs synthesized in this study were characterized by spectrophotometry, zeta potential, and high-resolution transmission electron microscopy (HRTEM). Finally, detection capacity of metallic ions in synthetic water samples was evaluated. It was proved that the methanolic extract of Aspergillus niger obtained under established conditions has the necessary components for both synthesis and coating of CdS-NPs, as well as detection of metallic ions because it was possible to synthesize CdS-NPs with a hexagonal crystalline structure with a length of 2.56 ± 0.50 nm which were able to detect Pb2+, Cr6+, and Fe3+ at pH 4 and Co2+ at pH 8.

Oil-removal enhancement in media with keratinous or chitinous wastes by hydrocarbon-degrading bacteria isolated from oil-polluted soils.

The aim of this work was to isolate oil-degrading bacteria that use chitin or keratin as carbon sources from oil contaminated soils; and additionally to study if oil removal by these bacteria is enhanced when a chitinous or a keratinous waste is added to the culture media. To isolate the above-mentioned bacteria, 12 soil samples were collected close to an oil-well. Such soils showed unsuitable nutrients content, but their counts of heterotrophic bacteria ranged within 10(5)-10(8) CFU g(-1) soil, of which 0.1-77% corresponded to oil hydrocarbon-degrading ones. By sampling on plates, 109 oil-degrading bacterial isolates were obtained. Their keratinase and chitinase activities were then screened by plate assays and spectrophotometric methods, resulting in 13 isolates that were used to integrate two mixed cultures, one keratinolytic and the other chitinolytic. These mixed cultures were grown in media with oil, or oil supplemented with chicken-feathers or shrimp wastes. The oil-hydrocarbon removal was measured by gas chromatography. Results showed that keratinolytic bacteria were better enzyme producers than the chitinolytic ones, and that oil removal in the presence of chicken-feathers was 3.8 times greater than with shrimp wastes, and almost twice, in comparison with oil-only added cultures. Identification of microorganisms from the mixed cultures by 16S rDNA, indicated the presence of seven different bacterial genera; Stenotrophomonas, Pseudomonas, Brevibacillus, Bacillus, Micrococcus, Lysobacter and Nocardiodes. These findings suggest that the isolated microorganisms and the chicken-feather wastes could be applied to the cleaning of oil-contaminated environments, whether in soil or water.

A field trial for an ex-situ bioremediation of a drilling mud-polluted site.

The remediation of drilling mud-polluted sites in the Southeast of Mexico is a top priority for Mexican oil industry. The objective of this work was to find a technology to remediate these sites. A field trial was performed by composting in biopiles, where four 1ton soil-biopiles were established, one treatment in triplicate and one unamended biopile. Amended biopiles were added with nutrients to get a C/N/P ratio of 100/3/0.5 plus a bulking agent (straw) at a soil/straw ratio of 97/3. Moisture content was maintained around 30-35%. Results showed that, after 180 d, total petroleum hydrocarbon (TPH) concentrations decreased from 99300+/-23000mgTPHkg(-1) soil to 5500+/-770mgTPHkg(-1) for amended biopiles and to 22900+/-7800mgTPHkg(-1) for unamended biopile. An undisturbed soil control showed no change in TPH concentrations. Gas chromatographic analysis showed residual alkyl dibenzothiophene type compounds. Highest bacterial counts were observed during the first 30 d which correlated with highest TPH removal, whereas fungal count increased at the end of the experimentation period. Results suggested an important role of the straw, nutrient addition and water content in stimulating aerobic microbial activity and thus hydrocarbon removal. This finding opens an opportunity to remediate old polluted sites with recalcitrant and high TPH concentration.

Colloidal chitin stained with Remazol Brilliant Blue R, a useful substrate to select chitinolytic microorganisms and to evaluate chitinases.

A simple and sensitive method based on the use of colloidal chitin stained with Remazol Brilliant Blue R (RBB) is proposed to evaluate chitinase activity. If this colloidal-stained substrate is included as a carbon source in a liquid medium, this technique allows the selection or the comparison of chitinolytic microorganisms. The colloidal substrate is proportionally solubilized and the dye released is spectrophotometrically quantified at 595 nm. The procedures used for the staining and fixing of RBB in the colloidal chitin, and a comparison with the commercial substrate chitin-azure, are presented. The influence of several physicochemical and enzymatic parameters on the release of dyes is also shown. Both stained substrates were used for studying the effect of pH, substrate concentration, temperature and time on the chitinase reaction of Bacillus thuringiensis Bt-107.

Isolation and characterization of bacteria degrading polychlorinated biphenyls from transformer oil.

Polychlorinated biphenyls from transformer oil were degraded in liquid culture under aerobic conditions using a mixed bacterial culture isolated from a transformer oil sample with a high content of polychlorinated biphenyls and other hydrocarbons. Four strains were identified, three of them corresponded to genus Bacillus, the other one to Erwinia. Bacteria in the transformer oil could remove as much as 65% of polychlorinated biphenyls (88% W/V in the transformer oil). Additional data showed that the two isolated strains of B. lentus were able to grow on transformer oil and degrade polychlorinated biphenyls by 80 and 83%. Our results provide evidence that microorganisms occurring in transformer oil have the potential to degrade polychlorinated biphenyls.

Chitosanase activity in Bacillus thuringiensis.

The ability to produce extracellular chitosanase (EC 3.2.1.132) was found by plate assays in 18 (23%) out of 77 crystalliferous strains of Bacillus thuringiensis. The best chitosanase producer was selected after the growth chosen in a liquid medium with colloidal chitosan as carbon source. Enzyme production was optimized (a 4-d incubation at 32 degrees C with shaking in a medium of pH 6.5 with 4% colloidal chitosan) and the enzyme was partially characterized. This is the first report on the chitosanase of B. thuringiensis.

Identification of recalcitrant hydrocarbons present in a drilling waste-polluted soil.

During spills of hydrocarbons in soil, it has been observed that aliphatic and the slightly aromatic hydrocarbons are first to be removed, however, branched aliphatic and aromatic compounds, polycyclic aromatic hydrocarbons (PAHs) and their similar heteroatoms with sulfur (PAS) remain strongly absorbed to soil particles. It is important to point out that studies of biodegradation of alkyl-substituted PAHs and PAS are scarce and most of them have been carried out using only available standard compounds. The aim of this investigation was to identify and to quantify the aliphatic, alkyl polycyclic aromatic, and sulfured recalcitrant fractions present in a contaminated soil with drilling wastes. A modified method of shaking-centrifugation extraction was implemented for the extraction of compounds from contaminated soil. The organic extract obtained was purified and fractionated using aluminum oxide. Gas Chromatograph with flame ionization detector (GC-FID) and Gas Chromatograph with mass spectrometer detector (GC-MS) identified the aliphatic, PAHs and PAS fractions. Hydrocarbon composition in the soil contaminated with 140,000 mg TPHs/Kg soil, consisted in 80% of branched aliphatic compounds of C10 to C22, 15% of alkyl PAHs, and 5% of PAS compounds. Lineal, lineal branched, and cyclic branched aliphatic hydrocarbons, as well as their alkyl naphthalene, anthracene and phenantrene, methyldibenzothiophene, dimethyldibenzothiophene, and dimethylnaphto[2,3-b]thiophene compounds were identified by CG-MS. The identification of compounds in soil P31, allowed us to speculate on the origin of the contamination and the natural attenuation that had occurred at this site.