Biomass and lipid enhancement in Chlorella sp. with emphasis on biodiesel quality assessment through detailed FAME signature.

In this study, the concentrations of MgSO4, salinity and light intensity were optimised for maximum biomass productivity and lipid content in Chlorella sp. Lipid synthesized at varied experimental conditions was also assessed in detail for biodiesel properties through FAME analysis. FAMEs mainly composed of C16:0, C16:1(9), C16:3(7, 10, 13), C18:0, C18:1(11), C18:2(9, 12), C18:3(9, 12, 15). The optimum biomass productivity (372.50mgL(-1)d(-1)) and lipid content (32.57%) was obtained at MgSO4-150ppm; salinity-12.5ppm, and light intensity-25µmolm(-2)s(-1). However, at this condition the cetane number, a major biodiesel property was not complying with worldwide biodiesel standard. Therefore, further optimisations were done to check the suitability of biodiesel fuel. The optimum biomass productivity (348.47mgL(-1)d(-1)) and lipid content (12.43%) with suitable biodiesel fuel properties was obtained at MgSO4-50ppm, salinity-25ppm and light intensity-100µmolm(-2)s(-1). The validation experiments confirmed the closeness of predicted and measured response values.

Stress-induced lipids are unsuitable as a direct biodiesel feedstock: a case study with Chlorella pyrenoidosa.

The effects of various stresses on the suitability of lipid synthesized by Chlorella pyrenoidosa for biodiesel production were investigated. Lipids were characterized for detailed fatty acid methyl ester profiling and biodiesel properties like cetane number (CN), iodine value, cold filter plugging point (CFPP). Maximum biomass productivity (106.63 mgL(-1)d(-1)) and lipid content (29.68%) were obtained at indoor cultivation (nitrate sufficient, pH 8-10, 24h illumination). However, compared to this condition, other nitrate sufficient cultures [pH 6-8 and 10-12 (24h illumination), and at ambient CO2 and 16:8h light:dark photoperiod (pH unadjusted)] showed ∼12-14% lower lipid productivity. Upon 50% nitrate depletion (at indoor and outdoor; pH unadjusted) lipid content has increased by 7.62% and 17%, respectively. Though stress conditions helped enhancing lipid accumulation, there was two-fold increase in PUFA content compared to that observed at pH 8-10. This resulted in fuel properties which did not comply with the biodiesel standards.

Cellular alterations and modulation of protein expression in bitumen-challenged human osteoblast cells.

PURPOSE: There are many arguments on the carcinogenic potential of bitumen extract. The mechanism of bitumen-induced damage is not well understood at the molecular level. Therefore, in the present study, cell-transforming and tumor-inducing potential of bitumen extract was studied using in vitro [human osteosarcoma (HOS) cells] and in vivo [nude and severe combined immunodeficiency (SCID) mice] models. METHODS: Gas chromatography/mass spectrometry (GC/MS) analysis was carried out to find out the existence of carcinogenic compounds in the bitumen extract. Cell transformation test, anchorage independence assay, karyotyping assay, tumorigenicity assay, and 2-DE analysis were used to find out the effect of bitumen using the in vitro and in vivo models. RESULTS: GC/MS analysis showed the existence of carcinogenic compounds in the bitumen extract. HOS cells were treated with different concentrations (25, 50, and 100 µl/ml) of bitumen extract. Compared to the parental HOS cells, bitumen transformants (HOS T1 and HOS T2) showed the characteristics of anchorage independency, chromosomal anomaly, and cellular transformation. Interestingly, bitumen transformants were not able to form tumor in nude/SCID mice. Proteomic analysis revealed the existence of 19 differentially expressed proteins involved in progression of cancer, angiogenesis, cell adhesion, etc. CONCLUSIONS: Exposure of bitumen extract to HOS cells results in the cellular transformation similar to cancer cells and can modulate proteins involved in the progression of cancer. We state that the non-tumorogenic potential of bitumen transformant in nude/SCID mice can be attributed to the downregulation of galectin-1, chromodomain helicase DNA-binding protein 1-like gene, and membrane-associated guanylate kinase 2 protein.

Genetic polymorphism of CYP2D6∗2 C→T 2850, GSTM1, NQO1 genes and their correlation with biomarkers in manganese miners of Central India.

Manganese (Mn) intoxication is most often regarded as an occupational manifestation and occurs in places such as manganese mines, dry cell battery plants and ceramic industries. In the present study, the influence of genetic polymorphism in cytochrome P450 2D6 (CYP2D6∗2), glutathione S-transferase M1 (GSTM1) and NAD(P)H quinone oxidoreductase 1 (NQO1) genes on blood manganese and plasma prolactin concentrations in manganese miners was investigated. Genotyping of CYP2D6∗2 C→T 2850 and NQO1 C→T 609 was carried out using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) whereas the genotyping of GSTM1 was carried out by multiplex PCR using beta globin as an internal control. Manganese miners with CYP2D6∗2 C→T 2850 variant genotype had relatively low Mn concentration [GM: 21.4±8.9 µg L(-1)] than the subjects with wild (GM: 36.3±8.5 µg L(-1)) and heterozygous (GM: 34.4±6.9 µg L(-1)) genotypes. Miners with CYP2D6∗2 variant genotypes showed low prolactin levels (GM: 13.13±1.6 ng mL(-1)) compared to the wild (GM: 16.4.4±1.5 µg L(-1)) and heterozygous (GM: 18.7±1.6 ng mL(-1)) genotypes. Gene-gene interaction studies also revealed that the subjects with CYP2D6∗2 C→T 2850 variant genotypes had low levels of Mn and prolactin. Our new findings suggest that CYP2D6∗2 C→T 2850 variant genotypes can regulate plasma prolactin levels in manganese miners of Central India and could be involved in the fast metabolism of blood manganese, compared to wild and heterozygous genotypes.

Gymnema montanum H. protects against alloxan-induced oxidative stress and apoptosis in pancreatic beta-cells.

The present study evaluated the molecular mechanism of antidiabetic property of G. montanum leaf extract (GLEt) against alloxan-induced apoptotic cell death in rat insulinoma cells (RINm5F). The pre-treatment of GLEt (5 microg and 10 microg/ml) resulted in significant decrease in intracellular Ca(2+) concentration, nitric oxide (NO) production along with increase in mitochondrial membrane potential in alloxan (7mM/ml) treated cells. Further GLEt reduced apoptosis by inhibiting the release of cytochrome c and subsequent cleavage of PARP and caspase-3. The immunochemical staining of 8-hydroxydeoxyguanosine (8-OHdG) also evidenced the suppression of oxidative stress by GLEt. The cell cycle analysis, annexin-V labelling assay and TUNEL assay showed the suppression of apoptosis by the treatment of GLEt. Moreover, GLEt significantly increased the cellular antioxidant levels and decreased the lipid peroxides in alloxan-treated RINm5F cells. Taken together, these findings suggest that G. montanum protects pancreatic beta-cells against reactive oxygen species (ROS) by counteracting with mitochondrial membrane permeability and inhibition of the apoptotic pathway.

Biological decolourization of C.I. Direct Black 38 by E. gallinarum.

In the present study, an Enterococcus gallinarum strain was isolated from effluent treatment plant of a textile industry based on its ability to decolourize C.I. Direct Black 38 (DB38), a benzidine-based azo dye. Effects of dye concentration and medium composition on dye decolourization were studied. The strain was found to decolourize DB38 even under aerobic conditions. Kinetics of DB38 decolourization was also examined, and V(max) and K(s) of decolourization were found to be higher in Luria broth (12.8 mg l(-1)h(-1) and 490.6 mg l(-1)) than in minimal medium (4.09 mg l(-1)h(-1) and 161.84 mg l(-1)). However, decolourization rate/biomass was found to be higher in minimal medium than in Luria broth, indicating greater decolourization efficiency of biomass in the former. The study also revealed biodegradation of DB38 to benzidine and its further deamination to 4-aminobiphenyl (4-ABP) by the culture. Ammonia released during this process was used as nitrogen source for growth of the culture.

Biodiversity and dye decolourization ability of an acclimatized textile sludge.

In the present study, sludge sample from biological treatment plant of a textile industry was acclimatized for decolourization of azo dye Direct Black 38 (DB38). A continuous culture experiment showed that the acclimatized sludge could decolourize 76% of 100mg/l DB38. Bacterial community in the sludge was analyzed using culture-independent molecular approach to get the complete picture of its diversity. RFLP analysis of its 16S rRNA gene library divided the clones into 14 distinct groups. Phylogenetic analysis of these groups showed that they belonged to five different bacterial lineages: beta- and gamma-Proteobacteria (3 and 4 respectively), Bacteroidetes (2), Firmicutes (4) and Actinobacteria (1). The largest number of clones was found to cluster in the gamma-Proteobacteria (54%), followed by Firmicutes (19%), beta-Proteobacteria (14%), Bacteroidetes (10%) and Actinobacteria (3%).

Azoreductase and dye detoxification activities of Bacillus velezensis strain AB.

Azo dyes are known to be a very important and widely used class of toxic and carcinogenic compounds. Although lot of research has been carried out for their removal from industrial effluents, very little attention is given to changes in their toxicity and mutagenicity during the treatment processes. Present investigation describes isolation of a Bacillus velezensis culture capable of degrading azo dye Direct Red 28 (DR28). Azoreductase enzyme was isolated from it, and its molecular weight was found to be 60 kDa. The enzyme required NADH as cofactor and was oxygen-insensitive. Toxicity and mutagenicity of the dye during biodegradation was monitored by using a battery of carefully selected in vitro tests. The culture was found to degrade DR28 to benzidine and 4-aminobiphenyl, both of which are potent mutagens. However, on longer incubation, both the compounds were degraded further, resulting in reduction in toxicity and mutagenicity of the dye. Thus, the culture seems to be a suitable candidate for further study for both decolourization and detoxification of azo dyes, resulting in their safe disposal.

Heavy metal status and oxidative stress in diesel engine tuning workers of central Indian population.

OBJECTIVE: To assess the oxidative stress induced due to heavy metal exposure. Exposed populations are selected from an engine tuning station and control from the same area with no occupational exposure. METHOD: Standard methods were followed for enzymatic assay, and heavy metals in blood and urine were analyzed by using inductively coupled plasma-optical emission spectrophotometer after microwave digestion. RESULT: Changes in mean blood Pb, Cd, and Ni concentrations in blood and urine of exposed population of all age groups (20 to 35, 35 to 45, and 46 to 58 years) and exposure durations (< or =10, 11 to 20, and >20 years) were statistically not significant. However, exposed workers exhibited statistically significant higher antioxidant status in terms of serum glutathione-S-transferase activity, malondialdehyde level, and catalase activity. CONCLUSION: The findings in this article suggest that occupational exposure to diesel exhaust of engine tuning workers causes induction of oxidative stress, which cannot be correlated with the heavy metals status in blood and urine of an exposed population.

Enrichment and isolation of endosulfan degrading and detoxifying bacteria.

In the present study, degradation of endosulfan by a mixed culture isolated from a pesticide-contaminated soil was studied in batch experiments. After two weeks of incubation, the mixed culture was able to degrade 73% and 81% of alpha and beta endosulfan respectively. Endodiol was identified by GC/MS as degradation intermediate. The toxicity studies of endosulfan before and after degradation were carried out using micronucleus assay on human polymorphonuclear cells. The findings suggested that the metabolism of endosulfan isomers by the mixed culture was accompanied by significant reduction in the toxicity. Studies were also carried out to quantify the degradation potential of the individual species in the mixed bacterial culture. Two cultures identified by 16S rRNA as Stenotrophomonas maltophilia and Rhodococcus erythropolis were found to be responsible for majority of the degradation by the mixed culture. S. maltophilia showed better degradation efficiency compared to that by R. erythropolis. This is the first report of endosulfan degradation using the above-mentioned organisms.

Kinetics of decolourisation and biotransformation of direct black 38 by C. hominis and P. stutzeri.

In the present study, a consortium of Cardiobacterium hominis and Pseudomonas stutzeri was isolated from an effluent treatment plant of a textile industry, based on its ability to decolourise azo dyes including direct black 38 (DB38), a benzidine-based azo dye. The role of each culture in the decolourisation process was elucidated, and C. hominis was found to decolourise the dye. Although P. stutzeri could not decolourise the dye, it was found to synergistically enhance dye decolourisation activity of C. hominis by scavenging oxygen in the medium and creating an anaerobic condition (oxidation/reduction potential -440 mV), which is known to be necessary for azo dye decolourisation. Together, the cultures could decolourise 90.5% of 100 mg l(-1) DB38 within 24 h. Kinetics of DB38 decolourisation was also examined, and P. stutzeri was found to increase V (max) and K (m) of decolourisation activity of C. hominis by 3.6- and 3-fold, respectively. The study also revealed a pathway of DB38 degradation with the release of benzidine from DB38 and subsequent degradation of benzidine to 4-aminobiphenyl by the cultures.

Decolorisation and detoxification of Direct Blue-15 by a bacterial consortium.

Studies were carried out on decolorisation and biotransformation of the dye Direct Blue-15 into 3,3'-dimethoxybenzidine (O'-dianisidine) and a sulphonated derivative by a five-member bacterial consortium. Chromatographic studies revealed further complete biodegradation of 3,3'-dimethoxybenzidine coupled with release of ammonia, but the recalcitrant sulphonated derivative persisted. The microorganisms identified in the mixed consortium by 16S rDNA sequence analysis were Alcaligenes faecalis, Sphingomonas sp. EBD, Bacillus subtilis, Bacillus thuringiensis and Enterobacter cancerogenus. The cytotoxicity data showed a significant reduction in the toxicity (P<0.001) of the degraded dye as evidenced from the number of viable human polymorphonuclear leukocyte cells present.