Growth responses and metal accumulation capabilities of woody plants during the phytoremediation of tannery sludge.

Five woody plants species (i.e. Terminalia arjuna, Prosopis juliflora, Populus alba, Eucalyptus tereticornis and Dendrocalamus strictus) were selected for phytoremediation and grow on tannery sludge dumps of Common Effluent Treatment Plant (CETP), Unnao (Uttar Pradesh), India. Concentration of toxic metals were observed high in the raw tannery sludge i.e. Fe-1667>Cr-628>Zn-592>Pb-427>Cu-354>Mn-210>Cd-125>Ni-76 mg kg(-1) dw, respectively. Besides, physico-chemical properties of the raw sludge represented the toxic nature to human health and may pose numerous risks to local environment. The growth performances of woody plants were assessed in terms of various growth parameters such as height, diameter at breast height (DBH) and canopy area of plants. All the plant species have the capabilities to accumulate substantial amount of toxic metals in their tissues during the remediation. The ratio of accumulated metals in the plants were found in the order Fe>Cr>Mn>Pb>Zn>Cu>Cd>Ni and significant changes in physico-chemical parameters of tannery sludge were observed after treatment. All the woody plants indicated high bioconcentration factor for different metals in the order Fe>Cr>Mn>Ni>Cd>Pb>Zn>Cu. After one year of phytoremediation, the level of toxic metals were removed from tannery sludge up to Cr (70.22)%, Ni (59.21)%, Cd (58.4)%, Fe (49.75)%, Mn (30.95)%, Zn (22.80)%, Cu (20.46)% and Pb (14.05)%, respectively.

Impact of phosphorous on biochemical changes in Hordeum vulgare L. in mixed cropping with chickpea.

Multiple cropping (i.e. intercropping or mixed cropping) plays an important role in agriculture because of the effective utilization of resources, significantly enhancing crop productivity compared with that of monocultured crops. The study was planed to assess the effect of various concentrations (00, 30, 60, 90 kg ha(-1)) of phosphorous on the biochemical composition of grains of Hordeum vulgare L. (NDB-1050) in mixed cropping system with Chickpea. Phosphorous is an essential ingredient for plants to convert atmospheric N (N2) into an ammonium (NH4) as a useable form. The available nitrogen content was found more in the year 2006 (131 kg ha(-1)) than year 2005 (105 kg ha(-1)). The results of available nitrogen content were showed that the mixed cropping system enhances N fixation process because phosphorous also influences nodule development through its basic functions in plants as an energy source. Reducing, non reducing and total sugar content of H. vulgare L. were influenced by changes in the phosphorous doses. Maximum protein (13.43%) was obtained at 60 kg P2O5 ha(-1) during the year 2006. Lysine, tryptophan and methionine content were found maximum in year 2006, respectively. Total mineral content of grains of plant (0.99 g 100g(-1)) was found maximum by the application of 60 kg P2O5 ha(-1). It is possible that there was an increase in the soil N made available by the leguminous chickpea species, and this could be another reason why there was an increase in Hordeum vulgare L. shoot mass per plant with intercropping with chickpea.

Involvement and interaction of microbial communities in the transformation and stabilization of chromium during the composting of tannery effluent treated biomass of Vallisneria spiralis L.

Tannery effluent treated with aquatic macrophyte Vallisneria spiralis L. for 14 d showed significant improvement in physico-chemical properties and reduction in Cr concentration. Accumulation of Cr was found maximum in roots (358 microg g(-1)dw) as compared to shoot (62 microg g(-1)dw) of the plant. A laboratory scale composter was designed with the objectives to investigate the physico-chemical changes and role of microbes in stabilization and transformation of Cr in the composting material. Results revealed that the composting process was quick within 7-21 d as indicated by peak time for various physico-chemical parameters and drop in C/N ratio up to acceptable limit. The profile of microbial communities indicated that population of anaerobic, aerobic and nitrifying bacteria increased quickly at the initial phase, and reached a peak level of 4.2 x 10(6), 9.78 x 10(8) and 9.32 x 10(9) CFU g(-1), respectively at 21 d; while population of actinomycetes and fungi was found maximum i.e. 3.29 x 10(7) and 9.7 x 10(6) CFU g(-1), respectively, after 35 d of composting. Overall bacterial population dominated over the actinomycetes and fungi during the composting process. Cr((VI)) was transformed to Cr((III)) due to the microbial activity during the process. Sequential extraction of Cr fractionation showed its stabilization via changing into organic matter-bound and residual fractions during the composting.

Responses of cyanobacterium Anabaena doliolum during nickel stress.

Growth and biochemical responses of heterocystous nitrogen fixing cyanobacteria Anabaena doliolum were studied upon exposure to various concentrations of nickel (0.1 to 100 microM) for duration of up to 4 days, in view of its tolerance. The growth of the cyanobacterium was increased in terms of cell density (O.D), heterocyst frequency chlorophyll-a, carotenoid and c-phycocynin up to moderate exposure (10 microM for 96 hr of Ni treatment). Protein and total non protein thiol were measured as stress responsive metabolites and frequency of heterocyst and spores were observed in responses to Ni. The present study demonstrated that the tested cyanobacterium exhibited dose specific responses of metal towards studied different morphological, physiological and biochemical parameters.

Screening and identification of early warning algal species for metal contamination in fresh water bodies polluted from point and non-point sources.

The water bodies of Lucknow, Unnao and Kanpur (U.P.), India polluted through various point and non point sources were found to be either eutrophic or oligotrophic in nature. These water bodies supported a great number of algal diversity, which varied seasonally depending upon the physico-chemical properties of water. Further, the water bodies polluted through non point sources supports diverse algal species, while the water bodies polluted through point sources supports growth of tolerant blue green algae. High biomass producing algal species growing in these water bodies have accumulated significant amount of metals in their tissues. Maximum amount of Fe was found accumulated by species of Oedogonium sp. II (20,523.00 microg g(-1) dw) and Spirogyra sp. I (4,520.00 microg g(-1) dw), while maximum Chromium (Cr) was found accumulated in Phormedium bohneri (2,109.00 microg g(-1) dw) followed by Oscillatoria nigra (1,957.88 microg g(-1) dw) and Oedogonium sp. I (156.00 microg g(-1) dw) and Ni in Ulothrix sp. (495.00 microg g(-1) dw). Results showed that some of these forms growing in polluted environment and accumulating high amounts of toxic metals may be used as bioindicator species, however, their performance in metal contaminated water under different ecological niche is to be ascertained.

Isolation and characterization of chromate resistant bacteria from tannery effluent.

The tannery effluent emanating from Common Effluent Treatment Plant (CETP), Unnao (U.P, India) was found toxic in nature, having high BOD, COD, TDS and Cr content (5.88 mg l(-1)), which supported growth of chromate tolerant bacteria. Several chromate tolerant bacteria have been isolated from these effluent and maximum tolerant four strains (NBRIP-1, NBRIP-2, NBRIP-3 and NBRIP-4) were characterized in this study. These strains showed multiple metal and antibiotic resistances. Growth of these strains was reduced at higher Cr concentration with extention of lag phase. Chromium accumlulation by these isolates may have a great potential in recovery and detoxification of Cr from tannery effluent.

Morphology and cultural behavior of Botryococcus protuberans with notes on the genus.

The green alga Botryococcus protuberans was isolated from its natural environment and its morphology under different cultural conditions was examined. The alga was characterized by a high starch content and reddish oil drops as the assimilatory products. Photosynthetic pigments, Chl a, Chl b, carotenoids and xanthophylls are present. Modification of environmental conditions in modified Chu-10 medium resulted in optimum growth of the alga. Fatty acid composition revealed palmitic acid being the major component, while lauric acid, myristic acid and stearic acid were found in less quantity.

Methaemoglobinaemia in epidemic proportions.

Five cases of methaemoglobinaemia, following exposure to industrial waste, are examined, investigated and managed successfully. The condition is discussed in detail.

Studies of the hepatic mitochondrial and microsomal mixed-function oxidase system during Plasmodium yoelii infection and inducer treatment in Swiss albino mice.

Plasmodium yoelii infection resulted in depression of hepatic mitochondrial and microsomal mixed-function oxidase system indices, e.g. cytochrome P-450, cytochrome b5 and phase II detoxification enzyme glutathione-S-transferase, while heam and haemozoin registered a marked increase in Swiss albino mice. Phenobarbitone (inducer) treatment showed induced levels of hepatic mitochondrial and microsomal cytochrome P-450 and glutathione-S-transferase in normal as well as in infected mice. The induced cytochrome P-450 and glutathione-S-transferase activities were similar in normal and infected mice. The findings were further supported by the isoenzymic profile and drug-binding properties of the terminal monoxygenase, cytochrome P-450.

Filarial DNA and its interaction with polyamines and antifilarial drugs.

The interaction of DNA from filarial parasite Setaria cervi with polyamines was monitored by melting temperature (Tm) profile, condensation and B to Z DNA transition and compared with DNA of Escherichia coli, Micrococcus luteus and calf thymus having different GC content. Polyamines, viz. spermine and spermidine, stabilized the secondary structure of all DNAs as indicated by increase in Tm value. UV absorption studies indicated B to Z DNA transition in the presence of polyamines. The amount of polyamines required for B to Z transition was dependent upon base composition of DNA and charge of the polyamine. Filarial DNA (AT rich) required six times higher concentration of spermine as compared to GC rich DNA for B to Z DNA transition. Spermidine was not effective in causing transition of S. cervi DNA even at Spd:DNA-P ratio of 20. The antifilarial compound suramin significantly decreased melting temperature of filarial DNA as compared to GC rich DNAs of other parasites. Suramin adversely affected condensation and B to Z DNA transition of various DNAs but prior addition of polyamines protected the DNAs from the destabilizing effect of suramin.

Identification of the polyamine N8-acetyltransferase involved in the pathway of 1,3-diaminopropane production in Acanthamoeba culbertsoni.

A cytosolic polyamine N-acetyltransferase that preferentially catalyzes the acetylation of spermidine in the N8-position was identified in the free-living pathogenic amoeba Acanthamoeba culbertsoni. In addition to spermidine, the enzyme also catalyzed the acetylation of spermine and putrescine with Michaelis constants (Km values) of 97, 12, and 10 microM, respectively. The Km value for acetylcoenzyme A (acetyl-CoA) was estimated to be 11 microM, whereas CoA had an inhibitory constant of 6 microM. The N-acetylase has a molecular mass of approximately 45 kDa. That the enzyme preferentially catalyzed the acetylation of spermidine at the N8-position, resulting in N8-acetylspermidine, the preferred substrate of the polyamine oxidase found in A. culbertsoni, indicates a role for the enzyme in the production of 1,3-diaminopropane, the major polyamine found in the Acanthamoeba.

In vitro susceptibility of Acanthamoeba culbertsoni to inhibitors of folate biosynthesis.

The effects of different sulphonamides, dihydrofolate reductase inhibitors and other inhibitors of folate metabolism on growth of Acanthamoeba culbertsoni in a chemically defined medium are reported. Among the sulphonamides, sulphamethoxazole and sulphadiazine were most effective followed by sulphanilamide and sulphaguanidine. Inhibition by each sulphonamide was reversed by p-aminobenzoic acid as well as folic acid. 7-Methylguanosine, a pteridine synthesis-inhibitor, did not inhibit multiplication of A. culbertsoni. Among the dihydrofolate reductase inhibitors, pyrimethamine blocked the amoebic growth at 100 micrograms/ml, while trimethoprim and cycloguanil palmoate failed to cause significant inhibition of growth even at 250 micrograms/ml. Metoprine inhibited amoebic growth completely at 50 micrograms/ml. Methotrexate and a thymidylate synthetase inhibitor 5-fluorouracil inhibited growth strongly, with IC50 values (the concentration of the drug which causes 50% inhibition of the growth at 72 h) of 1.97 and 2.45 micrograms/ml, respectively. Inhibition by methotrexate, metoprine or 5-fluorouracil could not be reversed by folic acid, folinic acid, thymidine, or folinic acid plus thymidine. The results indicate unusual features in A. culbertsoni folate metabolism.

Polyamine metabolism in Acanthamoeba culbertsoni.

1,3-Diaminopropane has been identified as the major polyamine of Acanthamoeba culbertsoni. N-acetylputrescine and spermidine were present in appreciable amounts and putrescine as well as N-acetylspermidine were also detected, but spermine was absent. Changes in polyamine levels were observed during the growth of amoebae. Ornithine decarboxylase activity was detected in cell-free extracts but there was very low activity of arginine and lysine decarboxylases. A potent polyamine oxidase was demonstrated which preferentially acted on N8-acetyl-spermidine as the substrate while N1-acetylspermidine was a poor substrate; free polyamines did not serve as a good substrate for this enzyme. Active uptake of polyamines by the amoebae was also demonstrated.

Characterization of Ca(2+)-ATPase of Setaria cervi (Nematoda: Filarioidea): effect of phenothiazines and anthelmintics.

1. A Mg2+ independent, Ca(2+)-ATPase requiring high concentrations of Ca2+ (5 mM) for the activation, equally distributed in cuticle-muscular-hypodermis, genital organs and gastrointestinal tissues and mainly localized in 10,000 g pellet fraction, was identified in Setaria cervi, a bovine filarial parasite. 2. Filarial enzyme showed Km value of 3.33 mM for ATP as computed from the double reciprocal Lineweaver-Burk plot. 3. The enzyme could be completely solubilized by sonication with about 4-fold increase in specific activity of the enzyme. 4. The enzyme showed about 2-fold activation by the calmodulin fractions isolated from S. cervi and rat brain homogenates. 5. The enzyme was highly sensitive to inhibition with some phenothiazine derivatives. Trifluoperazine was observed to be the most potent inhibitor followed by promethazine and chlorpromazine. 6. Some anthelmintics viz. diethycarbamazine and centperazine were found to be highly potent inhibitors of the enzyme, significant inhibition of filarial Ca(2+)-ATPase was also observed with levamisole and suramine. 7. Studies indicate Ca(2+)-ATPase of S. cervi as a potential chemotherapeutic target.

Polyamine oxidase from Acanthamoeba culbertsoni specific for N8-acetylspermidine.

Polyamine oxidase plays a key role in the catabolism of polyamines and regeneration of spermidine and putrescine. The mammalian enzyme utilises N1-acetylspermidine, and N8-acetylspermidine, although formed in the mammals, is not catabolised further. We have characterised an enzyme from Acanthamoeba culbertsoni which acts preferentially on N8-acetylspermidine. The highly unstable enzyme was stabilised in the presence of glycerol or dimethylsulphoxide together with spermine and purified 400-fold by a combination of DEAE-cellulose, CM-cellulose, spermine-Sepharose and Sephacryl S-300 chromatography. The enzyme has a pH optimum of 8 and a temperature optimum of 45 degrees C. The relative activities on different substrates are: N8-acetylspermidine 100%, N1-acetylspermine 40%, N1-acetylspermidine 1%, N1,8-diacetylspermidine 1% and N1,12-diacetylspermine 15%. Free polyamines and substrates of monoamine oxidase were not attacked. The enzyme yielded diaminopropane as an end product of catabolism and could be involved in the biosynthesis of this unusual polyamine present in large amounts in this organism.

Action of DL-alpha-difluoromethyl ornithine on Acanthamoeba culbertsoni.

The multiplication of A. culbertsoni in the peptone medium was not inhibited by 10-20 mM concentration of alpha-difluoromethyl ornithine (DMFO) while a partial and transient inhibition of cell multiplication was observed by 10-20 mM DFMO in proteose peptone, yeast extract, glucose (PYG) medium. Ornithine decarboxylase (ODC) activity in the cells and cell free extracts was strongly inhibited by DFMO, excluding enzyme refractoriness and impermeability of cells for DFMO as the possible causes of DFMO resistance. The presence of polyamines in the peptone and PYG media as well as uptake of polyamines by the amoebae has been demonstrated. The growth and multiplication of A. culbertsoni in chemically defined medium was not affected by 1-5 mM DFMO while 10-20 mM DMFO yielded partial inhibition. A lowering of diaminopropane levels and enhancement of spermidine levels was observed in DFMO inhibited cells and level of ODC was drastically reduced in the inhibited cultures. Uptake of polyamines from the growth media may partly account for DFMO resistance of A. culbertsoni. Alternative mechanisms for DFMO resistance are indicated.