Plant and yeast consortium for efficient remediation of dyes and effluents: a biochemical and toxicological study.

Textile effluent carries a range of dyes that may be recalcitrant and resistant to biodegradation. A unique consortium of the Fimbristylis dichotoma and Saccharomyces cerevisiae is exploited for the biodegradation of an azo dye Rubine GFL and actual textile effluent. This consortium enhances the rate of biodegradation of Rubine GFL and actual textile effluent with an excellent rate of biodegradation of 92% for Rubine GFL and 68% for actual textile effluent when compared to the individual one within 96 h. Speedy decolorization of Rubine GFL and actual textile effluent was observed due to the induction of oxido-reductive enzymes of the FD-SC consortium. Along with the significant reduction in the values of COD, BOD, ADMI, TSS, and TDS with 70, 64, 65, 41, and 52%, respectively, in experimental sets treated with FD-SC consortium. The biodegradation of Rubine GFL was confirmed with UV-Vis spectroscopy at the preliminary level, and then, metabolites formed after degradation were detected and identified by FTIR, HPLC, and GC-MS techniques. Also, decolorization of the dye was observed in the sections of the root cortex of Fimbristylis dichotoma. The toxicity of dye and metabolites formed after degradation was assessed by seed germination and bacterial count assay, where increased germination % and bacterial count from 31×107CFUs to 92 × 107 CFUs reflect the nontoxic nature of metabolites. Furthermore, the nontoxic nature of metabolites was confirmed by fish toxicity on Cirrhinus mrigala showed normal structures of fish gills and liver in the groups treated with FD-SC consortium proving the better tactic for biodegradation of dyes and textile effluent.

Construction and implementation of floating wetpark as effective constructed wetland for industrial textile wastewater treatment.

Fimbristylis dichotoma, Ipomoea aquatica, Pluchea tomentosa and their co-plantation (consortium FIP) autonomously degrade Orange 3R. Consortium FIP showed 84% removal of Orange 3R within 48 h, which is a higher dye elimination rate than individual plant systems. Oxidoreductase enzymes like tyrosinase (76%), varatryal alcohol oxidase (85%), lignin peroxidase (150%), riboflavin reductase (151%), laccase (171%), NADH-DCIP reductase (11%) and azo reductase (241%) were expressed in consortia FIP during Orange 3R degradation. UV-vis spectroscopy, enzyme activities, HPTLC, FTIR and GC-MS confirmed mineralization of Orange 3R into its metabolites. Microscopic investigation of root tissue revealed the harsh effect of dye on root tissues. Toxicity assessment on the HepG2 cell line demonstrated the toxic nature of Orange 3R, which gets reduced after phyto-treatment with consortia FIP. Floating wetpark of consortia FIP was found more efficient for the treatment of industrial textile waste and accomplished 87%, 86%, 75%, 49% and 46% removal of COD, BOD, color, TSS and TDS of effluent.

In situ textile wastewater treatment in high rate transpiration system furrows planted with aquatic macrophytes and floating phytobeds.

Plants are known to remediate dyes, metals and emerging contaminants from wastewaters. Vetiveria zizanioides, a perennial bunchgrass showed removal of Remazol Red (RR, 100 mg/L) up to 93% within 40 h. Root and shoot tissues of V. zizanioides revealed induction in dye degrading enzymes viz. lignin peroxidase by 2.28 and 1.43, veratryl alcohol oxidase 2.72 and 1.60, laccase 6.15 and 3.55, and azo reductase 2.17 and 2.65-fold, respectively, during RR decolorization. Substantial increase was observed in the contents of chlorophyll a, chlorophyll b, and carotenoids in the plant leaves during treatment. Anatomical studies of roots, HPLC and GC-MS analysis of metabolites, and phytotoxicity assessment confirmed phytotransformation of RR into nontoxic metabolites. Floating phytobed with V. zizanioides treated textile wastewater (400 L) effectively and reduced ADMI, COD, BOD, TDS, and TSS by 74, 74, 81, 66 and 47%, respectively within 72 h. In-situ treatment of textile wastewater for 5 days in constructed furrows planted with semiaquatic plants, V. zizanioides, Ipomoea aquatica and its consortium-VI decreased ADMI by 68, 61 and 76%, COD by 75, 74 and 79%, BOD by 73, 71 and 84%, TDS by 77, 75 and 83%, and TSS by 34, 31 and 51%, respectively. This treatment was also useful to remove arsenic, cadmium, chromium and lead from wastewater. Overall observation suggests wise strategy to use this plantation in the furrows of high rate transpiration system and phytobeds in deep water for textile wastewater treatment.

In situ phytoremediation of dyes from textile wastewater using garden ornamental plants, effect on soil quality and plant growth.

In situ phytoremediation of dyes from textile wastewater was carried out in a high rate transpiration system ridges (91.4 m × 1.0 m) cultivated independently with Tagetes patula, Aster amellus, Portulaca grandiflora and Gaillardia grandiflora which reduced American Dye Manufacturers Institute color value by 59, 50, 46 and 73%, respectively within 30 d compared to dye accumulated in unplanted ridges. Significant increase in microbial count and electric conductivity of soil was observed during phytoremediation. Reduction in the contents of macro (N, P, K and C), micro (B, Cu, Fe and Mn) elements and heavy metals (Cd, As, Pb and Cr) was observed in the soil from planted ridges due to phyto-treatment. Root tissues of these plants showed significant increase in the specific activities of oxido-reductive enzymes such as lignin peroxidase, laccase, veratryl alcohol oxidase, tyrosinase and azo reductase during decolorization of textile dyes from soil. Anatomical studies of plants roots revealed the occurrence of textile dyes in tissues and subsequent degradation. A minor decrease in plant growth was also observed. Overall surveillance suggests that the use of garden ornamental plants on the ridges of constructed wetland for the treatment of dyes from wastewater along with the consortia of soil microbial flora is a wise and aesthetically pleasant strategy.

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

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 60h at a concentration of 50ppm, while 95% dye removal was achieved by consortium FA within 48h. 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.

Co-plantation of aquatic macrophytes Typha angustifolia and Paspalum scrobiculatum for effective treatment of textile industry effluent.

Field treatment of textile industry effluent was carried out in constructed drenches (91.4m×1.2m×0.6m; 65.8m3) planted independently with Typha angustifolia, Paspalum scrobiculatum and their co-plantation (consortium-TP). The in situ treatment of effluent by T. angustifolia, P. scrobiculatum and consortium-TP was found to decrease ADMI color value by 62, 59 and 76%, COD by 65, 63 and 70%, BOD by 68, 63 and 75%, TDS by 45, 39 and 57%, and TSS by 35, 31 and 47%, respectively within 96h. Heavy metals such as arsenic, cadmium, chromium and lead were also removed up to 28-77% after phytoremediation. T. angustifolia and P. scrobiculatum showed removal of Congo Red (100mg/L) up to 80 and 73%, respectively within 48h while consortium-TP achieved 94% decolorization. Root tissues of T. angustifolia and P. scrobiculatum revealed inductions in the activities of oxido-reductive enzymes such as lignin peroxidase (193 and 32%), veratryl alcohol oxidase (823 and 460%), laccase (492 and 182%) and azo reductase (248 and 83%), respectively during decolorization of Congo Red. Anatomical studies of roots, FTIR, HPLC, UV-vis Spectroscopy and GC-MS analysis verified the phytotransformation. Phytotoxicity studies confirmed reduced toxicity of the metabolites of Congo Red.

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.

Ipomoea hederifolia rooted soil bed and Ipomoea aquatica rhizofiltration coupled phytoreactors for efficient treatment of textile wastewater.

Ipomoea aquatica, a macrophyte was found to degrade a highly sulfonated and diazo textile dye Brown 5R up to 94% within 72 h at a concentration of 200 mg L(-1). Induction in the activities of enzymes such as azoreductase, lignin peroxidase, laccase, DCIP reductase, tyrosinase, veratryl alcohol oxidase, catalase and superoxide dismutase was observed in leaf and root tissue in response to Brown 5R exposure. There was significant reduction in contents of chlorophyll a (25%), chlorophyll b (17%) and carotenoids (30%) in the leaves of plants. HPLC, FTIR, UV-vis spectrophotometric and HPTLC analyses confirmed the biotransformation and removal of parent dye from solution. Enzymes activities and GC-MS analysis of degradation products lead to the proposal of a possible pathway of phytotransformation of dye. The proposed pathway of dye metabolism revealed the formation of Napthalene-1,2-diamine and methylbenzene. Toxicity study on HepG2 cell lines showed a 3 fold decrease in toxicity of Brown 5R after phytoremediation by I. aquatica. Hydrophytic nature of I. aquatica leads to its exploration in a combinatorial phytoreactor with Ipomoea hederifolia soil bed system. Rhizofiltration with I. aquatica and soil bed treatment by I. hederifolia treated 510 L of effluent effectively within 72 h. I. aquatica along with I. hederifolia could decolorize textile industry effluent within 72 h of treatment as evident from the significant reductions in the values of COD, BOD, solids and ADMI. Further on field trials of treatment of textile wastewater was successfully carried out in a constructed lagoon.

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.

Analysis of genetic variability in endemic medicinal plants of genus Chlorophytum from the Indian subcontinent using amplified fragment length polymorphism marker.

The genus Chlorophytum consists of medicinally important species like Chlorophytum borivilianum, C. tuberosum and C. attenuatum. Uncontrolled harvest of this plant from wild habitat due to its high commercial value made the species of this genus be listed in the Red Data Book of Indian plants as an endangered species. In India, approximately nineteen species of Chlorophytum are found; out of these, only C. borivilianum is cultivated commercially. The objective of this study was to measure genetic diversity, population structure and phylogenetic relationship among the species using Amplified Fragment Length Polymorphisms (AFLP). Fifteen pairs of primer (out of 64 primer pairs screened) were used to analyse the genetic diversity in eighteen species of genus Chlorophytum. Cluster analysis, estimation of the gene flow among the species and of the phylogeographic distribution of this genus were carried out using an AFLP data matrix. A high level of genetic diversity was observed on the basis of the percentage of polymorphic bands (99.91%), Shannon's information index (0.3592) and Nei's gene diversity (0.2085) at species level. Cluster analysis of UPGMA dendrogram, principal component analysis and Bayesian method analysis resolved these species in three different clusters, which was supported by morphological information. The Mantel test (r=0.4432) revealed a significant positive correlation between genetic and geographic distances. The collected data have an important implication in the identification, authentication, and conservation of the species of the genus Chlorophytum.

Phytoremediation of sulfonated Remazol Red dye and textile effluents by Alternanthera philoxeroides: An anatomical, enzymatic and pilot scale study.

Alternanthera philoxeroides Griseb. a macrophyte was found to degrade a highly sulfonated textile dye Remazol Red (RR) completely within 72 h at a concentration of 70 mg L(-1). An induction in the activities of azoreductase and riboflavin reductase was observed in root and stem tissues; while the activities of lignin peroxidase, laccase and DCIP reductase were induced in leaf tissues. Some enzymes namely tyrosinase, veratryl alcohol oxidase, catalase and superoxide dismutase displayed an increase in their activity in all the tissues in response of 72 h exposure to Remazol Red. There was a marginal reduction in contents of chlorophyll a (20%), chlorophyll b (5%) and carotenoids (16%) in the leaves when compared to control plants. A detailed anatomical study of the stem during uptake and treatment revealed a stepwise mechanism of dye degradation. UV-vis spectrophotometric and high performance thin layer chromatographic analyses confirmed the removal of parent dye from solution. Based on the enzymes activities and gas chromatography-mass spectroscopic analysis of degradation products, a possible pathway of phytotransformation of RR was proposed which revealed the formation of 4-(phenylamino)-1,3,5-triazin-2-ol, naphthalene-1-ol and 3-(ethylsulfonyl)phenol. Toxicity study on Devario aequipinnatus fishes showed that the anatomy of gills of fishes exposed to A. philoxeroides treated RR was largely protected. The plants were further explored for rhizofiltration experiments in a pilot scale reactor. A. philoxeroides could decolorize textile industry effluent of varying pH within 96 h of treatment which was evident from the significant reductions in the values of American dye manufacturers' institute color, chemical oxygen demand, biological oxygen demand, total dissolved and total suspended solids.