Biodegradation of high molecular weight PAHs using isolated yeast mixtures: application of meta-genomic methods for community structure analyses

Bioaugmentation for the removal of polyaromatic hydrocarbons (PAHs) from wastewater using bacteria and yeasts is considered environment-friendly and a cost-effective technique. The effectiveness of this biodegradation system depends on the stability of inoculated microorganisms and the availability of nutrients. This study is aimed to investigate the removal of high molecular weight (HMW)-PAHs from biologically treated produced water using different biological systems. Three systems, inoculated with activated sludge (AS), the mixture of five yeast strains (MY), and the mixture of AS and the five yeast strains (SY), respectively, were constructed, and their performance for the removal of HMW-PAHs was compared over 10 weeks. The effluent of the biologically treated produced water from an oilfield was used as the influent after chrysene and benzo(a)pyrene were spiked as HMW-PAHs. Polymerase chain reaction-based denaturing gradient gel electrophoresis (PCR-DGGE) and fluorescent in situ hybridization (FISH) techniques were used to examine the changes in the structures and abundances of the bacterial and yeast communities in these three systems. Only SY and MY systems were capable to remove chrysene (90.7 % and 98.5 %, respectively) and benzo(a)pyrene (80.7 % and 95.2 %, respectively). PCR-DGGE analysis confirmed that all of the five yeast strains inoculated remained in the SY and MY systems, while FISH results showed that the relative abundance of yeast in the SY and MY systems (10.6 % to 21.9 %, respectively) were significantly higher than AS system (2.3 % to 7.8 %, respectively). The relative abundances of the catechol 2,3-dioxygenase (C23O) indicated that the copy number ratios of benzene ring cleavage gene C23O in the yeast amended systems were much higher than that in the AS system. In this study, all of the three systems were effective in removing the low molecular weight (LMW)-PAHs, while HMW-PAHs including chrysene and benzo(a)pyrene were efficiently removed by MY and SY systems, not by AS system. The high HMW-PAHs removal in the MY and SY bioaugmentation systems possibly attributed to the inoculation of the mixed yeast culture. By combining the PCR-DGGE results with the FISH analyses, it was found that yeast probably consisting mainly of the five inoculated strains inhabited in the two bioaugmentation systems as a dominant population. The relatively higher performance of the SY system might be attributed to the suspended growth type which permitted a more efficient contact between microbial cells and contaminants. The bioaugmentation systems (SY and MY) were successfully established by inoculating with five nonindigenous yeast strains and demonstrated high performance in removal of HMW-PAHs.     

Purification and Properties of Extracellular Poly(3-hydroxybutyrate) Depolymerase Produced by <Emphasis Type="Italic">Aspergillus fumigatus</Emphasis> 202

An extracellular poly(3-hydroxybutyrate) (PHB) depolymerase produced by a thermotolerant fungal soil isolate, Aspergillus fumigatus 202, was purified and characterized. Maximum PHB depolymerase production was obtained at the end of 48 h with initial medium pH 7.0 and 45 °C in Bushnell Haas Minerals medium containing PHB as sole source of carbon. The PHB depolymerase was purified using size exclusion chromatography to a fold purification of 20.62 and 61.62% yield. SDS-PAGE and isoelectric focusing revealed the molecular weight and pI of the purified enzyme as 63,744 Da and 4.2, respectively. N-terminal amino acid sequence of purified enzyme was HAXDAYLVK. This non-glycosylated enzyme was most active at pH 9.0 and 45 °C. Purified enzyme was inactivated by N-bromosuccinimide and dithiothreitol suggesting the involvement of tryptophan residues and disulfide bonds at its active site. Nonionic detergents like Tween 20, Tween 80 and Triton X-100 inhibited the enzyme activity. Ions like Ca⁺² and Mg⁺² (5 mM) increased the enzyme activity 1.5 times. Fe⁺² effectively inhibited the enzyme activity to 88% whereas Hg⁺² completely inhibited the enzyme.     

Development of Thixotropic Coatings Using Dimerized Soybean Oil Fatty Acids: The Case of Polyamide of 1,2-Phenylene Diamine

Soybean oil (SBO) was dimerized and the crude dimer acid product reacted with 1,2-phenylene diamine at 210 ± 5 °C under inert atmosphere to obtain fatty polyamide (FPA). The FPA was used to modify a commercial alkyd resin by reacting a mixture of the alkyd resin with 5 wt% of FPA at 120 °C for 80 min under inert atmosphere. The FTIR spectrum of the FPA modified resin showed evidence of higher degree of H-bonding than was found for the unmodified alkyd. White gloss coatings of 15, 20, 25, and 30% solids were formulated from the modified and unmodified resins and examined for performance with respect to: leveling, sag resistance, drying time, pigment settling, skinning tendency and film hardness. Results showed that the unmodified alkyd coatings exhibited good leveling but poor sag resistance at all solid contents. In contrast, FPA modified alkyd coatings combined good leveling with high sag resistance indicating their thixotropic nature. A strong tendency to pigment settling was observed for unmodified alkyd coatings but was not observed in the FPA modified alkyd coatings. The modified alkyd coatings showed skinning while the unmodified alkyd coatings did not skin. A 30% solids coating formulation of the FPA modified resin showed shorter surface dry time but longer hard dry time than the unmodified alkyd resin coating.     

Block Copolymers Containing (R)-3-Hydroxybutyrate and Isosorbide Succinate or (S)-Lactic Acid Mers

A new aliphatic block copolyester was synthesized in bulk from transesterification techniques between poly((R)-3-hydroxybutyrate) (PHB) and poly(isosorbide succinate) (PIS). Additionally, other two block copolyesters were synthesized in bulk either from transesterification reactions involving PHB and poly(l-lactide) (PLLA) or from ring-opening copolymerization of l-lactide and hydroxyl-terminated PHB, as result of a previous transesterification reactions with isosorbide. Two-component blends of PHB and PIS or PLLA were also prepared as comparative systems. SEC, MALDI-TOF mass spectrometry (MALDI-TOFMS), ¹H and ¹³C NMR spectroscopy, WAXD, solubility tests, and TG thermal analysis were used for characterization. The block copolymer structures of the products were evidenced by MALDI-TOFMS, ¹³C NMR, and WAXD data. The block copolymers and the corresponding binary blends presented different solubility properties, as revealed by solubility tests. Although the incorporation of PIS sequences into PHB main backbone did not enhance the thermal stability of the product, it reduced its crystallinity, which could be advantageous for faster biodegradation rate. These products, composed of PHB and PIS or PLLA sequences, are an interesting alternative in biomedical applications.     

Annual nutrients budget for the grazed and ungrazed sites of an alpine expanse in North-West Himalaya,India

This work was undertaken to analyze nutrient contents of vegetation in an alpine meadow—Tungnath, North-West Himalaya, India. The study pertains to the uptake, transfer and release of four main macronutrients (organic carbon, total nitrogen, total potassium and total phosphorus) in grazed (exposed to extensive grazing by cattles) and ungrazed (grazing completely prohibited) communities. Mineral concentration was recorded higher for the ungrazed sites compared to the grazed sites, and maximum standing state of nutrients was found in roots. Belowground compartment (roots) contributed maximum share of mineral elements to soil. Litter nutrients release was low because of low microbial activity and continuous removal of phytomass. Observations reveal that there was very little amount of nutrient release from phytomass and vegetation in alpine are very poor source of mineral recycling. Low transfer rate of minerals from one compartment to other is adequate for greater amount of these minerals that are translocated back into the storage organs. A small proportion get removed through rain splash or through the removal of hay during grazing as relatively high release rates in ungrazed sites when compared to grazed sites was observed. This translocation can be considered as an important adaptation in alpine plants for survival during adverse environmental conditions, against all types of biotic pressures and also for regeneration in the forthcoming growing season.     

An assessment of ecosystem services of Corbett Tiger Reserve,India

This paper examines the economic value of selected ecosystem services of Corbett Tiger Reserve, India. The direct cost was derived from secondary sources, and indirect and opportunity costs through socioeconomic surveys. For recreational value the individual approach to travel cost method was used, and to assess carbon sequestration the replacement cost method was used. The maintenance cost of the reserve was estimated as US $2,153,174.3 year⁻¹. The indirect costs in terms of crop and livestock depredation by wild animals ranged from US $2,153,174.3 year⁻¹. The indirect costs in terms of crop and livestock depredation by wild animals ranged from US 2,408 to US $37,958 village⁻¹ over a period of 5 years. The dependence of local communities was for fuel wood (US $37,958 village⁻¹ over a period of 5 years. The dependence of local communities was for fuel wood (US 7,346 day⁻¹), fodder (US $5,290 day⁻¹), small timber, and other nontimber forest products. The recreational value of the reserve was estimated as US $5,290 day⁻¹), small timber, and other nontimber forest products. The recreational value of the reserve was estimated as US 167,619 year⁻¹. With the cost per visitor being US $2.5, the consumers’ surplus was large, showing the willingness of visitors to pay for wildlife recreation. The forests of the reserve mitigate carbon worth US $2.5, the consumers’ surplus was large, showing the willingness of visitors to pay for wildlife recreation. The forests of the reserve mitigate carbon worth US 63.6 million, with an annual flow of US $65.0 ha⁻¹ year⁻¹. The other benefits of the reserve include US $65.0 ha⁻¹ year⁻¹. The other benefits of the reserve include US 41 million through generation of electricity since 1972. The analysis reveals that, though the benefits outweigh costs, they need to be accrued to local communities so as to balance the distribution of benefits and costs.     

Gender,age and subject matter: impact on teachers’ ecological values

By means of environmental education, teachers are supposed to intervene in pupils’ awareness and concern of environmental issues. Teachers’ environmental attitudes, however, have rarely been studied. We analysed 367 German pre- and in-service teachers’ environmental attitudes for their structure and socio-economic correlates within the frame of a European project. Thus, we retrieved for adults the two-dimensional model of ecological values (2-MEV) presented for adolescents by Bogner and Wiseman (Sci Educ Int 15(1):27–48, 2004). For the two domains, Preservation and Utilisation, the psychometric structure was confirmed and discriminative correlations with age, gender and teaching subject were unveiled. Further research is needed to deduce the implications for teaching practice.     

Evaluation of the gene expression changes in Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) as affected by the bio-removal of toxic textile dyes from aqueous solution in small-scale bioreactor

A laboratory-scale bioremediation unit was designed, built and tested for the bio-removal of several Direct textile dyes. Four experiments were carried out to assess the efficiency of the bioremediation unit using Aspergillus niger fungal strain. Three commonly used Direct dyes and textile dyes mixture (simulated effluent: Direct brown, Direct violet, Direct green) were tested in this study. The strain of A. niger was efficient in the removal of the three Direct dyes. The decolorization percentages of the dyes after 24 h of incubation were 56.2, 51.7, and 95.4% for Direct brown, Direct green, Direct violet dyes, respectively. The percentages increased up to 79.4, 86.4, and 96.7% after 72 h of incubation for the same dyes, respectively. The results also showed that the fungal strain reduced the chemical oxygen demand values of simulated dye effluents from 165 to 564 mg/l with most of the dyes. The assessment of bioremediation products on biomodel was conducted using a fresh water fish. The liver and brain of Nile tilapia were tested to evaluate the expression of genes coding for several proteins related to stress such as metallothioneins (MTs), cytochrome P450 (CYP450), and heat shock proteins (HSPs). To assess the alterations in the gene expression, ten animals from each group were killed after 4 weeks of treatment. The results revealed significant increases in the brain and hepatic mRNA levels of all stress protein genes MT, CYP450, Hsp70a, b, and Hsp47 in the fish groups treated with industrial Direct violet, green, and brown dye water. Exposure of tilapia to bioremediation products after treatment with A. niger fungi reduced the over-expression of the stress protein genes in the brain and liver tissues.