Micronucleus induction by oxidative metabolites of trichloroethylene in cultured human peripheral blood lymphocytes: a comparative genotoxicity study

The genotoxic effects of oxidative metabolites of trichloroethylene (TCE), namely chloral hydrate, trichloroacetic acid (TCA), dichloroacetic acid (DCA), and trichloroethanol (TCEOH) were examined in human peripheral blood lymphocytes. In this context, lymphocytes were exposed in vitro to 25, 50, and 100 μg/ml concentrations of these metabolites separately for a period of 48 h and examined for micronucleus (MN) induction through flow cytometer. At 50 μg/ml TCE metabolites, TCA (6.33?±?0.56 %), DCA (5.06?±?0.55), and TCEOH (4.70?±?1.73) induced highly significant (p?<?0.001) frequency of MN in comparison to control (1.03?±?0.40) suggestive of their genotoxic potential. However, exposure of 100 μg/ml of all the metabolites consistently declined the frequencies of MN which in some cases was equable to that of observed at 25 μg/ml. Further, cytotoxicity and cell cycle disturbances were also measured to find out the association of these endpoints with the MN induction. DNA content analysis revealed 3–4-fold elevation of S-phase at all the concentrations tested. Particularly, at 100 μg/ml, treatment elevation of S-phase was significantly (p?<?0.0001) higher as compared to the control. Present findings together with earlier reports indicate that TCE induces genotoxicity through its metabolites. Interaction of these metabolites with DNA, as evident by elevated S-phase, seems to be the major cause of MN induction. However, involvement of spindle disruption cannot be ruled out. This comparative study also suggests that after TCE exposure, the metabolic efficiency of human to generate oxidative metabolites determines the extent of genotoxicity.     

Carbon, nitrogen, and phosphorus budget in scampi (Macrobrachium rosenbergii) culture ponds

Experiments were conducted for the study of nutrient budget in ten farmer's ponds (0.2–0.5 ha) in Orissa, India with a mean water depth of 1.0–1.2 m. Scampi (Macrobrachium rosenbergii) were stocked in these ponds at stocking density of 3.75–5.0/m². The average initial body weight of scampi was 0.02 mg. The culture period was for 4 months. Feed was the main input. Total feed applied to these ponds ranged from 945 to 2261 kg pond/cycle (crop). The feed conversion ratio varied 1.65 to 1.78. In addition to feed, rice straw, urea, and single super phosphate were applied to these ponds in small amounts for plankton production. At harvest time, the average weight of scampi varied from 60–90 g. The budget showed that feed was the major input of nitrogen (N), phosphorus (P), and carbon in these ponds. The inorganic fertilizer (urea and single super phosphate), organic fertilizer (rice straw and yeast extract), and inlet water, either from the initial fills or from rainwater, were the source of all other N, P, and organic carbon (OC) to these ponds. Total N applied to these ponds through all these inputs ranged from 44.45 to 103.98 kg N per crop, 12.23 to 28.79 kg P per crop, and from 381.54 to 905.22 kg OC per crop, respectively. Among all the inputs, feed alone accounted for 95.34 % N, 97.98 % P, and 94.27 % OC, respectively. Recovery of 16.34 to 38.66 kg N (average 29.27 kg), 1.28 to 3.02 kg P (average 2.29 kg), and 63.21 to 149.51 kg OC (average 113.20 kg), respectively, by the scampi harvest were observed in these ponds. Thus, harvest of scampi accounted for recovery of 35.18 to 39.01 (average 36.85 %) of added N, 10.09 to 10.97 (average 10.44 %) of added P, and 7.57 to 17.12 (average 16.34 %) of added OC, respectively.     

Assessment of denitrification gaseous end-products in the soil profile under two water table management practices using repeated measures analysis

The denitrification process and nitrous oxide (N2O) production in the soil profile are poorly documented because most research into denitrification has concentrated on the upper soil layer (0-0.15 m). This study, undertaken during the 1999 and 2000 growing seasons, was designed to examine the effects of water table management (WTM), nitrogen (N) application rate, and depth (0.15, 0.30, and 0.45 m) on soil denitrification end-products (N2O and N2) from a corn (Zea mays L.) field. Water table management treatments were free drainage (FD) with open drains and subirrigation (SI) with a target water table depth of 0.6 m. Fertility treatments (ammonium nitrate) were 120 kg N ha(-1) (N120) and 200 kg N ha(-1) (N200). During both growing seasons greater denitrification rates were measured in SI than in FD, particularly in the surface soil (0-0.15 m) and at the intermediate (0.15-0.30 m) soil depths under N200 treatment. Greater denitrification rates under the SI treatment, however, were not accompanied with greater N2O production. The decrease in N2O production under SI was probably caused by a more complete reduction of N2O to N2, which resulted in lower N2O to (N2O + N2) ratios. Denitrification rate, N2O production and N2O to (N2O + N2) ratios were only minimally affected by N treatments, irrespective of sampling date and soil depth. Overall, half of the denitrification occurred at the 0.15- to 0.30- and 0.30- to 0.45-m soil layers, and under SI, regardless of fertility treatment level. Consequently, sampling of the 0- to 0.15-m soil layer alone may not give an accurate estimation of denitrification losses under SI practice.     

Toxicokinetics,metabolism, and microsomal studies of chlorpropham in rats

A study on the toxicokinetic behavior, metabolism of chlorpropham, and its effect on cytochrome P₄₅₀ from liver microsomes was carried out in albino rats after a single and consecutive oral administration at 500?mg?kg^?1 body weight for 10 and 20 days. Chlorpropham was detected in the blood at 0.08?h (11.43?±?1.72?µg?mL^?1) reaching a maximum concentration at 2?h (30.90?±?2.55?µg?mL^?1) and a minimum at 48?h (1.95?±?0.20?µg?mL^?1) after a single oral administration of 500?mg?kg^?1. The absorption rate constant (K _a) was 0.66?±?0.48?h^?1. The Vd _area (18.01?±?2.78?L?kg^?1) and t _1/2 β (12.23?±?1.96?h) values suggested a wide distribution and long persistence of the compound in the body, respectively. The higher Cl_R (0.82?±?0.00?L?kg^?1?h^?1) compared to Cl_H (0.18?±?0.02?L?kg^?1?h^?1) value indicated that a major portion of chlorpropham was excreted through the urine (30%) compared to the faeces (2.81%). Chlorpropham residue was detected in all tissues of rat at 0.25?h while its metabolite, meta-chloroaniline was detected in liver, kidney, heart, lung, and spleen tissue at 0.25?h. Meta-chloroaniline was not detected in skeletal muscle, brain, fat, and stomach tissue at any time of the observation period. Maximum concentrations of chlorpropham and meta-chloroaniline were detected at 2?h (except in the spleen), and minimum concentrations of chlorpropham at 24 (heart, lung, spleen, skeletal muscle, and stomach) and 48?h (liver, kidney, brain, and fat tissue) respectively; and meta-chloroaniline at 24?h (except heart and spleen). The tissue half-life of chlorpropham in rat varied from 3.80 to 11.60?h. Repeated oral administration of chlorpropham at 500?mg?kg^?1 for 10 and 20 days caused an induction of the liver microsomal pellet of rat.     

Chemometric evaluation of heavy metal pollutions in Patna region of the Ganges alluvial plain,India: implication for source apportionment and health risk assessment

While metal pollution and distribution in soil are well documented for many countries, the situation is more serious in developing countries because of the rapid increase in industrialization and urbanization during last decades. Although it is well documented in developed countries, data about substantial metal pollution in Indian soil, especially in eastern Ganges alluvial plain (GAP), are limited. In this study, eight different blocks of Patna district located in eastern GAP were selected to investigate the contamination, accumulation, and sources of metals in surface soil considering different land use types. Additionally, human health risk assessment was estimated to mark the potential carcinogenic and non-carcinogenic effect of metals in soil. The concentration of all metals (except Pb) in soil was below the Indian standard limit of the potential toxic element for agricultural soil. Pb was the most abundant in soil, followed by Zn and Cu, and accounted for 52, 33 and 8% of the total metal. In terms of land use types, roadside soil detected higher concentrations of all metals, followed by park/grassland soil. Principal component analysis results indicated traffic pollution and industrial emissions are the major sources of heavy metals in soil. This was further confirmed by strong inter-correlation of heavy metals (Cd, Cr, Ni, Cu and Pb). Human health risk assessment results indicated ingestion via soil as the primary pathway of heavy metal exposure to both adults and children population. The estimated hazard index was highest for Pb, suggesting significant non-carcinogenic effect to both adults and children population. The children were more prone to the non-carcinogenic effect of Pb than adults. However, relatively low cancer risk value estimated for all metals suggested non-significant carcinogenic risk in the soil.     

Studies on mycorrhizal inoculation on dry matter yield and root colonization of some medicinal plants grown in stress and forest soils

Five medicinal plants viz. Abelmoschatus moschatus Linn., Clitoria tematea L., Plumbagozeylanica L., Psorolea corylifolia L. and Withania sominifera L. were grown in a polypot experiment in five soils representing coal mine soil, coppermine soil, fly ash, skeletal soil and forest soil with and without mycorrhizal inoculations in a completely randomized block design. Dry matter yield and mycorrhizal root colonization of plants varied both in uninoculated and inoculated conditions. The forest soil rendered highest dry matter due to higher yield of A. moschatus, P. zeylanica and P corylifolia while fly ash showed lowest dry matter without any inoculants. P. cematea were best in coalmine soil and W. sominifera in copper mine soil without mycorrhizal inoculation. The mycorrhiza was found to enhance the dry matter yield. This contributed minimum 0.19% to maximum up to 422.0% in different soils as compared to uninoculated plants. The mycorrhizal dependency was noticed maximum in plants grown in fly ash followed by coal mine soil, copper mine soil, skeletal soil and forest soil. The mycorrhizal response was increased maximum in W. sominifera due to survival in fly ash after inoculation followed by P corylifolia and P cematea. Percent root colonization in inoculated plant was increased minimum of 1.10 fold to maximum of 12.0 folds in comparison to un-inoculated plants . The native mycorrhiza fungi were also observed to colonize 4.0 to 32.0% roots in plants understudy. This study suggests that mycorrhizal inoculation increased the dry matter yield of medicinal plants in all soils under study. It also helps in survival of W. sominifera in fly ash.     

Identification of species-specific RAPD markers in genus Cenchrus

Cenchrus is an important component of major grass cover of world. Similar to the other major tropical grasses most of the species in genus Cenchrus are also apomictic in nature hence correct and precise identification of accessions and species are problematic and dubious. In the present study 187 decamer oligonucleotide primers were tested for PCR-based DNA amplification of six prominent species of genus Cenchrus. Of these, 32 potential repetitive and polymorphic primers were tested for identification of species-specific markers for C. ciliaris, C. setigerus, C. pennisetiformis, C. prieurri, C. biflorus and C. myosuroides. These primers yielded 51 unique RAPD markers either specific to a species (37) or shared by two or more species (14). Maximum markers were shared between C. ciliaris and C. setigerus confirming theirmore closeness to each other Primers like OPF09, OPF11, OPR15, OPAJ11, OPQ10 and OPAK20 generated strong intense bands can be used on priority in identifying the species from their natural habitat for the development of species-specific core germplasm. Due to apomictic nature this is the prime method of developing cultivars, as morphological characters are largely unable to distinguish them. The level of variation observed clearly suggest RAPD as an appropriate marker for genetic studies and in identifying the lines with species-specific markers for Cenchrus germplasm management activity and also maintaining identity and purity for proprietary reasons.     

Use of degenerate primers in rapid generation of microsatellite markers in Panicum maximum

Guineagrass (Panicum maximum Jacq.) is an important forage grass of tropical and semi-tropical regions, largely apomictic and predominantly exist in tetraploid form. For molecular breeding work, it is prerequisite to develop and design molecular markers for characterization of genotypes, development of linkage map and marker assisted selection. Hence, it is an important researchable issue to develop molecular markers in those crops where such information is scanty. Among many molecular markers, microsatellites or simple sequence repeat (SSR) markers are preferred markers in plant breeding. Degenerate primers bearing simple sequence repeat as anchor motifs can be utilized in rapid development of SSR markers; however selection of suitable degenerate primers is a prerequisite for such procedure so that SSR enriched genomic library can be made rapidly. In the present study seven degenerated primers namely KKVRVRV(AG)10, KKVRVRV(GGT)5, KKVRVRV(CT)10, KKVRVRV(AAT)6, KKVRVRV(GTG)6, KKVRVRV(GACA)5, and KKVRVRV(CAA)6 were used in amplification of Panicum maximum genomic DNA. Primers with repeat motifs (GGT)5 and (AAT)6 have not reacted whereas (AG)10, (GACA)5 and (CAA)6 highly informative as they have generated many DNA fragments ranging from 250 to 1600 bps as revealed from the results obtained with restriction digestion of recombinant plasmids. Primer with (CT)10 anchor repeat, amplified fragments of high molecular weight where as (GTG)6 primer generated only six bands with low concentration indicating less suitability of these primerin SSR markers development in P maximum.