Swapnil M. Patil

Low cost CaCl2 pretreatment of sugarcane bagasse for enhancement of textile dyes adsorption and subsequent biodegradation of adsorbed dyes under solid state fermentation

Pretreatments to sugarcane bagasse (SCB) such as CaCl2, alkali, ammonia, steam and milling showed 91%, 46%, 47%, 42% and 56% adsorption of Solvent Red 5B (SR5B); 92%, 57%, 58%, 56% and 68% adsorption of simulated dyes mixture (SDM), and 86%, 45%, 49%, 44% and 56% adsorption of a real textile effluent (RTE), respectively. However, the untreated SCB showed 32%, 38% and 30% adsorption of SR5B, SDM and RTE, respectively. Adsorption of SR5B on CaCl2 pretreated SCB follows pseudo-second order kinetics. SEM and FTIR analysis reveals the delignification of CaCl2 pretreated SCB. SR5B, SDM and RTE adsorbed on CaCl2, alkali, ammonia, steam and milling pretreated SCB were decolorized under solid state fermentation using isolated Providencia staurti strain EbtSPG. Tray bioreactor study showed 86% American Dye Manufacturers Institute (ADMI) removal of RTE in 72h. Biodegradation of adsorbed SR5B was confirmed using FTIR, HPLC and HPTLC.

Enhanced phytotransformation of Navy Blue RX dye by Petunia grandiflora Juss. with augmentation of rhizospheric Bacillus pumilus strain PgJ and subsequent toxicity analysis.

This study reveals the beneficial synergistic phytoremediation potential of Petunia grandiflora Juss. with its rhizospheric bacterial isolate Bacillus pumilus strain PgJ to decolorize reactive Navy Blue RX (NBRX) dye by their active enzymatic machinery. In vitro cultures of P. grandiflora and B. pumilus gave 80.01% and 76.80% while their consortium decolorized NBRX up to 96.86% within 36 h. Significant induction in the enzyme activities of lignin peroxidase (207%), tyrosinase (133%), laccase (161%), riboflavin reductase (78%) were seen in the roots of tissue cultured plants while enzymes tyrosinase (660%), laccase (689%), riboflavin reductase (528%) were induced significantly in the B. pumilus cells. Metabolites of treated NBRX were analyzed using UV-vis spectroscopy, gas chromatography and biotransformation was visualized using high performance thin layer chromatography profile. Metabolites of the dye exhibited reduced phytotoxicity Sorghum vulgare and Phaeseolus mungo and significant reduction in cytogenotoxicity on Allium cepa roots when compared to NBRX.

Efficient decolorization and detoxification of textile industry effluent by Salvinia molesta in lagoon treatment

Salvinia molesta, an aquatic fern was observed to have a potential of degrading azo dye Rubine GFL up to 97% at a concentration of 100mg/L within 72h using 60±2g of root biomass. Both root as well as stem tissues showed induction in activities of the enzymes such as lignin peroxidase, veratryl alcohol oxidase, laccase, tyrosinase, catalase, DCIP reductase and superoxide dismutase during decolorization of Rubine GFL. FTIR, GC-MS, HPLC and UV-visible spectrophotometric analysis confirmed phytotransformation of the model dye into smaller molecules. Analysis of metabolites revealed breakdown of an azo bond of Rubine GFL by the action of lignin peroxidase and laccase and formation of 2-methyl-4-nitroaniline and N-methylbenzene-1, 4-diamine. Anatomical tracing of dye in the stem of S. molesta confirmed the presence of dye in tissues and subsequent removal after 48h of treatment. The concentration of chlorophyll pigments like chlorophyll a, chlorophyll b and carotenoid was observed during the treatment. Toxicity analysis on seeds of Triticum aestivum and Phaseolus mungo revealed the decreased toxicity of dye metabolites. In situ treatment of a real textile effluent was further monitored in a constructed lagoon of the dimensions of 7m×5m×2m (total surface area 35m(2)) using S. molesta for 192h. This large scale treatment was found to significantly reduce the values of COD, BOD5 and ADMI by 76%, 82% and 81% considering initial values 1185, 1440mg/L and 950 units, respectively.

Bioreactor with Ipomoea hederifolia adventitious roots and its endophyte Cladosporium cladosporioides for textile dye degradation.

In vitro grown untransformed adventitious roots (AR) culture of Ipomoea hederifolia and its endophytic fungus (EF) Cladosporium cladosporioides decolorized Navy Blue HE2R (NB-HE2R) at a concentration of 20 ppm up to 83.3 and 65%, respectively within 96h. Whereas the AR-EF consortium decolorized the dye more efficiently and gave 97% removal within 36h. Significant inductions in the enzyme activities of lignin peroxidase, tyrosinase and laccase were observed in roots, while enzymes like tyrosinase, laccase and riboflavin reductase activities were induced in EF. Metabolites of dye were analyzed using UV-vis spectroscopy, FTIR and gas chromatography-mass spectrometry. Possible metabolic pathways of NB-HE2R were proposed with AR, EF and AR-EF systems independently. Looking at the superior efficacy of AR-EF system, a rhizoreactor was developed for the treatment of NB-HE2R at a concentration of 1000 ppm. Control reactor systems with independently grown AR and EF gave 94 and 85% NB-HE2R removal, respectively within 36h. The AR-EF rhizoreactor, however, gave 97% decolorization. The endophyte colonization additionally increased root and shoot lengths of candidate plants through mutualism. Combined bioreactor strategies can be effectively used for future eco-friendly remediation purposes.

Production of camptothecine using whey by an endophytic fungus: standardization using response surface methodology

Fusarium oxysporum kolhapuriensis, a novel endophytic fungi isolated from Nothapodytes nimmoniana Mabb. Grahm, was found to produce camptothecine (CPT) using whey as a complex medium. The highest production of CPT was attained using statistical methods Response Surface Methodology (RSM). Central Composite Design (CCD) was used to optimize the complex medium and culture conditions for maximum production of CPT by the fungus. The optimized medium that yielded 283 ± 0.27 mg l−1 of CPT contained 70% (v/v) of acid whey and 2% (w/v) malt extract. The other two culture parameters optimized through RSM were temperature (30 °C) and period of incubation (6 days). The production of CPT was confirmed by analytical techniques such as HPTLC, HPLC and LC-HRMS. This cost effective optimized medium using RSM might be useful for large scale CPT production which will ultimately reduce the further downstream processing cost.

Molecular characterization of intergeneric hybrid between Aspergillus oryzae and Trichoderma harzianum by protoplast fusion

Protoplast fusion between Aspergillus oryzae and Trichoderma harzianum and application of fusant in degradation of shellfish waste.

Phylogenetic analysis, genetic diversity and relationships between the recently segregated species of Corynandra and Cleoserrata from the genus Cleome using DNA barcoding and molecular markers

Cleome is the largest genus in the family Cleomaceae and it is known for its various medicinal properties. Recently, some species from the Cleome genus (Cleome viscosa, Cleome chelidonii, Cleome felina and Cleome speciosa) are split into genera Corynandra (Corynandra viscosa, Corynandra chelidonii, Corynandra felina), and Cleoserrata (Cleoserrata speciosa). The objective of this study was to obtain DNA barcodes for these species for their accurate identification and determining phylogenetic relationships. Out of 10 screened barcoding regions, rbcL, matK and ITS1 regions showed higher PCR efficiency and sequencing success. This study added matK, rbcL and ITS1 barcodes for the identification of Corynandra chelidonii, Corynandra felina, Cleome simplicifolia and Cleome aspera species in existing barcode data. Corynandra chelidonii and Corynandra felina species belong to the Corynandra genus, but they are not grouped with the Corynandra viscosa species, however clustered with the Cleome species. Molecular marker analysis showed 100% polymorphism among the studied plant samples. Diversity indices for molecular markers were ranged from He=0.1115-0.1714 and I=0.2268-0.2700, which indicates a significant amount of genetic diversity among studied species. Discrimination of the Cleome and Corynandra species from Cleoserrata speciosa was obtained by two RAPD primers (OPA-4 and RAPD-17) and two ISSR primers (ISSR-1 and ISSR-2). RAPD and ISSR markers are useful for the genetic characterization of these studied species. The present investigation will be helpful to understand the relationships of Cleome lineages with Corynandra and Cleoserrata species.

Phytobeds with Fimbristylis dichotoma and Ammannia baccifera for treatment of real textile effluent: An in situ treatment, anatomical studies and toxicity evaluation

Abstract Fimbristylis dichotoma, Ammannia baccifera and their co‐plantation consortium FA independently degraded Methyl Orange, simulated dye mixture and real textile effluent. Wild plants of F. dichotoma and A. baccifera with equal biomass showed 91% and 89% decolorization of Methyl Orange within 60 h at a concentration of 50 ppm, while 95% dye removal was achieved by consortium FA within 48 h. Floating phyto‐beds with co‐plantation (F. dichotoma and A. baccifera) for the treatment of real textile effluent in a constructed wetland was observed to be more efficient and achieved 79%, 72%, 77%, 66% and 56% reductions in ADMI color value, COD, BOD, TDS and TSS of textile effluent, respectively. HPTLC, GC‐MS, FTIR, UV–vis spectroscopy and activated oxido‐reductive enzyme activities confirmed the phytotrasformation of parent dye in to new metabolites. T‐RFLP analysis of rhizospheric bacteria of F. dichotoma, A. baccifera and consortium FA revealed the presence of 88, 98 and 223 genera which could have been involved in dye removal. Toxicity evaluation of products formed after phytotransformation of Methyl Orange by consortium FA on bivalves Lamellidens marginalis revealed less damage of the gills architecture when analyzed histologically. Toxicity measurement by Random Amplification of Polymorphic DNA (RAPD) technique revealed bivalve DNA banding pattern in treated Methyl Orange sample suggesting less toxic nature of phytotransformed dye products. Graphical abstract Figure. No Caption available. HighlightsF. dichotoma L. and A. baccifera L. decolorized Methyl Orange and real textile dye effluent.Co‐plantation of F. dichotoma L. and A. baccifera L. gave more efficient dye removal.Possible degradation pathways of Methyl Orange by all three systems are proposed.Effluents were treated note‐worthily in floating phyto‐beds by plants.Toxicity study on bivalve revealed less toxic nature of dye products.

Superparamagnetic core/shell nanostructures for magnetic isolation and enrichment of DNA

Fe3O4 magnetic nanoparticles (MNPs) are promising candidates for various biomedical applications due to their extraordinary properties. Such MNPs, surface modified with chitosan–glutaraldehyde (Fe3O4–CH/GLD) i.e. magnetic core/shell nanostructures, were used in the present study to investigate isolation and enrichment of bacterial DNA. Isolation was carried out in comparison with an organic method. FTIR was used to confirm biding of DNA onto the surface of the core–shell structures. The concentration of isolated DNA (yield) was 14.90 and 17.55 μg mL−1 for phenol/chloroform and magnetic isolation methods, respectively. The purity of isolated DNA was found to be 1.69 and 1.71 for phenol/chloroform and magnetic isolation methods, respectively. The present study firstly reports the comparison between magnetic and organic isolation of DNA. From both results (yield and purity), it was found that magnetic isolation of DNA was superior to the general organic method used for bacterial DNA isolation. Experiments for DNA enrichment were performed in batch mode and the effects of core/shell concentration, pH of the sample solution and temperature were optimized. The formation energy required for adsorption of DNA was found to be −55.56 × 10−23 J per molecule (−34.70 × 10−4 eV per molecule). The negative value indicates energy was utilized (endothermic process) for the adsorption of DNA onto the magnetic core/shells. The magnetic isolation method used in the present study was simple, fast, robust and ecofriendly (it does not require organic solvents or sophisticated equipment).

Relative profile analysis of molecular markers for identification and genetic discrimination of loaches (Pisces, Nemacheilidae).

Genus Nemacheilus, Nemachilichthys and Schistura belong to the family Nemacheilidae of the order Cypriniformes. The present investigation was undertaken to observe genetic diversity, phylogenetic relationship and to develop a molecular-based tool for taxonomic identification. For this purpose, four different types of molecular markers were utilized in which 29 random amplified polymorphic DNA (RAPD), 25 inter-simple sequence repeat (ISSR) markers, and 10 amplified fragment length polymorphism (AFLP) marker sets were screened and mitochondrial COI gene was sequenced. This study added COI barcodes for the identification of Nemacheilus anguilla, Nemachilichthys rueppelli and Schistura denisoni. RAPD showed higher polymorphism (100%) than the ISSR (93.75-100%) and AFLP (93.86-98.96%). The polymorphic information content (PIC), heterozygosity, multiplex ratio, and gene diversity was observed highest for AFLP primers, whereas the major allele frequency was observed higher for RAPD (0.5556) and lowest for AFLP (0.1667). The COI region of all individuals was successfully amplified and sequenced, which gave a 100% species resolution.