Detecting diclofenac in livestock carcasses in India with an ELISA: a tool to prevent widespread vulture poisoning

Diclofenac, a non-steroidal anti-inflammatory drug (NSAID), has caused catastrophic vulture declines across the Indian sub-continent. Here, an indirect ELISA is used to detect and quantify diclofenac in 1251 liver samples from livestock carcasses collected across India between August 2007 and June 2008, one to two years after a ban on diclofenac manufacture and distribution for veterinary use was implemented. The ELISAs applicability was authenticated with independent data obtained using LC-ESI/MS. Of 1251 samples, 1150 (91.9%) were negative for diclofenac using both methods, and 60 (4.8%) were positive at 10-4348 and 10-4441 μg kg(-1) when analysed by ELISA and LC-ESI/MS, respectively. The residue level relationship in the 60 positive samples was highly significant (p < 0.001, r(2) = 0.644). Data suggest that this immunological assay could be used not only for cost effective sample screening, but also for residue level semi-quantification.     

Lead tolerance and physiological adaptation mechanism in roots of accumulating and non-accumulating ecotypes of Sedum alfredii

Background, aim and scope

Lead (Pb) accumulation in soils affects plants primarily through their root systems. The aim of this study was to investigate early symptoms of the loss of membrane integrity and lipid peroxidation in root tissues and physiological adaptation mechanism to Pb in accumulating ecotypes (AE) and non-accumulating ecotypes (NAE) of Sedum alfredii under Pb stress in hydroponics.

Methods and results

Histochemical in situ analyses, fluorescence imaging, and normal physiological analysis were used in this study. Pb accumulation in roots of both AE and NAE increased linearly with increasing Pb levels (0?C200???M), and a significant difference between both ecotypes was noted. Both loss of plasma membrane integrity and lipid peroxidation in root tissues became serious with increasing Pb levels, maximum tolerable Pb level was 25 and 100???M for NAE and AE, respectively. Pb supplied at a toxic level caused a burst of reactive oxygen species (ROS) in root cells in both ecotypes. However, the root cells of AE had inherently higher activities of superoxide dismutase (SOD), guaiacol peroxidase (POD), and lipoxygenase (LOX) in control plants, and the induction response of these antioxidant enzymes occurred at lower Pb level in AE than NAE. AE plants maintained higher ascorbic acid and H₂O₂ concentrations in root cells than NAE when exposed to different Pb levels, and Pb induced more increase in dehydroascorbate (DHA), catalase (CAT), and ascorbate peroxidase (APX) in AE than NAE roots.

Discussion and conclusion

Results indicate that histochemical in situ analyses of root cell death and lipid peroxidation under Pb short-term stress was sensitive, reliable, and fast. Higher tolerance in roots of accumulating ecotype under Pb stress did depend on effective free oxygen scavenging by making complex function of both constitutively higher activities and sensitive induction of key antioxidant enzymes in root cells of S. alfredii.     

Decay profile and metabolic pathways of quinalphos in water, soil and plants

The widespread occurrence of pesticide residues in different agricultural and food commodities has raised concern among the environmentalists and food chemists. In order to keep a proper track of these materials, studies on their decay profiles in the various segments of ecosystem under varying environmental conditions are needed. In view of this, the metabolites of quinalphos in water and soil under controlled conditions and in plants, namely tomato and radish in field conditions have been analysed and possible pathways suggested. In order to follow the decay of the pesticide, an HPLC procedure has been developed. Studies conducted in water at different temperatures, pH and organic content reveal that the persistence of the pesticide decreases with the increase in all the three variables. In the three different types of soils studied, the effect of pH is more or less apparent on a similar line. On an average a faster decay is observed in the case of plants than in water and soil. The decay profiles in all these cases follow first order kinetics. The metabolites were identified by GC-MS. The investigations reflect that degradation occurs through hydrolysis, S-oxidation, dealkylation and thiono-thiol rearrangement. The pathways seem to be complex and different metabolites were observed with the change in the matrix. Quinalphos oxon, O-ethyl-O-quinoxalin-2-yl phosphoric acid, 2-hydroxy quinoxaline and quinoxaline-2-thiol were observed in all the matrices. Results further indicate that the metabolites, 2-hydroxy quinoxaline and oxon, which are more toxic than parent compound, persist for a longer time.     

Antioxidant properties and instrumental quality characteristics of spray dried Pangasius visceral protein hydrolysate prepared by chemical and enzymatic methods

Fish protein hydrolysates are digested form of protein with various bioactive properties where, the cleavages of molecular bonds of proteins can be broken by the enzymatic and chemical process. In this study, antioxidant properties of spray dried protein hydrolysate prepared from Pangasius viscera by using enzymatic (papain and pepsin), and chemical methods (hydrochloric acid and sodium hydroxide) were evaluated. Among the different treatments, pepsin-derived visceral protein hydrolysate showed the maximum antioxidant activity when used at higher concentrations. Essential amino acids (EAA) and hydrophobic amino acids are higher in papain-derived visceral protein hydrolysate. In pepsin-derived visceral protein hydrolysate, major proportion was contributed by glycine (Gly), glutamine (Glu), proline (Pro), and asparagine (Asp). Higher amount of aromatic amino acids are found in alkali-derived FVPH. Scanning electron microscopy (SEM) images of pepsin fish visceral protein hydrolysate showed better globular structure than the other treatments. It can be concluded that among the different treatments, the visceral protein hydrolysate prepared with pepsin had better overall quality regarding antioxidant properties and papain in nutritional point of view.

     

Chemical fractionation and heavy metal accumulation in the plant of Sesamum indicum (L.) var. T55 grown on soil amended with tannery sludge: Selection of single extractants

A pot experiment was carried out to study the single and sequential extractions of metals in different tannery sludge amendment and the potential of the plant of Sesamum indicum L. var. T55 (sesame) for the removal of metals from tannery waste contaminated site. The metal extraction efficiency obtained with each extractants was slightly different and follow the order; EDTA>DTPA>NH(4)NO(3)>NaNO(3)>CaCl(2). The correlation analysis between extractable metals in the different amendments of sludge and metal accumulation in the plant (lower and upper parts) showed better correlation for most of the tested metals with EDTA extraction. In this study, a sequential extraction technique was applied on different amendments of tannery sludge. The results showed that Mn, Zn, Cr and Cd were mostly associated with Fe-Mn oxide fraction in most of the amendments, K and Ni was found in residual (RES) fraction, Fe and Cu was bound with organic matter (OM) and RES fractions and Na was associated with carbonate (CAR) fraction. The metal accumulation after 60 d of growth of the plant was found in the order of K>Na>Fe>Zn>Cr>Mn>Cu>Pb>Ni>Cd and its translocation was found less in upper part. The accumulation of toxic metals (Cr, Ni and Cd) in the plants was found to increase with increase in sludge ratio, in contrast, the accumulation of Pb decreased. In view of growth parameters and metal accumulation in the plant, it was observed that lower amendments (25%) of tannery sludge were found suitable for the phytoremediation of most of the studied metals.     

Changes in amino acid profile and metal content in seeds of Cicer arietinum L. (chickpea) grown under various fly-ash amendments

Seeds of Cicer arietinum L. plants are edible and a valuable source of protein. Accumulation of toxic metals in the edible part of the plant, grown in fields close to fly-ash (FA) landfills, may pose a threat to human health. In the present study, the effects of FA and its amendments with different ameliorants viz., garden soil (GS), press mud (PM) and saw dust (SD), on total soluble protein contents, amino acid composition and metal accumulation in seeds were investigated in var. CSG-8962 and var. C-235 of C. arietinum. Plants accumulated adequate amounts of essential metals viz. Fe, Cu, Zn in seeds, while the toxic metals such as Cd and Cr were taken up in smaller quantities. The accumulation of Cr and Cd was less in var. C-235 than var. CSG-8962. Amendment of FA with PM enhanced the amount of soluble protein and amino acids in both varieties and was found to be superior among all tested ameliorants. Both quantitative and qualitative analysis of amino acids showed better response in var. C-235 as compared to var. CSG-8962. Thus var. C-235 seems to be suitable for cultivation in FA contaminated areas due to more accumulation of essential metals and less accumulation of toxic metals in seeds. Application of PM may further improve the growth of plants and nutritional quality of seeds.     

Bagasse-assisted bioremediation of ammonia from shrimp farm wastewater.

Development of new economically feasible ecofriendly products from agricultural wastes or byproducts for shrimp farm wastewater treatment is the objective of our continued research. Ammonia is a nitrogenous toxicant, which is commonly found in wastewater from shrimp farms. In the present study, we explored the possibility of the use of simply and inexpensively prepared bagasse products so that this abundant crop byproduct could be used to remove ammonia from shrimp farm wastewater. Bagasse, a natural highly fibrous lignocellulosic byproduct of sugarcane, was converted into five different products. Experimental results have shown that ammonia is efficiently removed from wastewater by four bagasse products with a dose of 1 to 6 g/L within 24 hours. The effect of bagasse products on other water quality parameters and growth kinetics of biofilm bacteria onto bagasse fiber have also been studied. Efficacies of products were compared by using statistical analysis. Products developed from bagasse are useful and economical.     

Disentangling horizontal pay dispersion: Experimental evidence

Although horizontal pay dispersion has been explored extensively using cross‐sectional field methods, it has received little attention using the control available through experimental designs. Many of the questions relevant to pay dispersion research can be addressed by taking an individual‐level experimental approach because this allows for clean separation of pay policies and individual effects. In this paper, we hypothesize both the motivation‐based and affect‐based effects of pay dispersion policies and test our hypotheses with a sample of over 400 participants in a real‐pay, real‐effort experiment. Results of the experiment provide support that two pay dispersion‐related pay policies, performance‐based pay dispersion and allocation criteria, have unique effects. Whereas motivation and performance effects are direct, pay satisfaction and interest in continuing work effects are the result of interactions, incorporating the performance level of workers. In follow‐up analyses, we find evidence that the temporal nature of responses to pay dispersion should be incorporated into future studies of pay dispersion.     

Investigation of low-level <Superscript>242</Superscript>Pu contamination on nutrition disturbance and oxidative stress in <Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.

Plutonium associated with higher molecular weight molecules is presumed to be poorly mobile and hardly plant available. In our present study, we investigate the uptake and effects of Pu treatments on Solanum tuberosum plants in amended Hoagland medium at concentrations of [²⁴²Pu] = 100 and 500 nm, respectively. We found a direct proof of oxidative stress in the plants caused by these rather low concentrations. For the confirmation of oxidative stress, we explored the production of nitric oxide (NO) and hydrogen peroxide (H₂O₂) by epifluorescence microscopy. Oxidative stress markers like lipid peroxidation and superoxide radicals (O₂ ^??) are monitored through histochemical analysis. The biochemical parameters i.e. chlorophyll and carotenoids are measured as an indicator of cellular damage in the tested plants including the enzymatic parameters such as catalase and glutathione reductase. From our work, we conclude that Pu in low concentration has no significant effects on the uptake of many trace and macroelements. In contrast, the content of O₂ ^?? , malondialdehyde (MDA), and H₂O₂ increases with increasing Pu concentration in the solution, while the opposite effects was found for NO, catalase, and glutathione reductase. These findings prove that even low concentration of Pu regulates ROS production and generate oxidative stress in S. tuberosum L.     

QSAR modeling for predicting mutagenic toxicity of diverse chemicals for regulatory purposes

The safety assessment process of chemicals requires information on their mutagenic potential. The experimental determination of mutagenicity of a large number of chemicals is tedious and time and cost intensive, thus compelling for alternative methods. We have established local and global QSAR models for discriminating low and high mutagenic compounds and predicting their mutagenic activity in a quantitative manner in Salmonella typhimurium (TA) bacterial strains (TA98 and TA100). The decision treeboost (DTB)-based classification QSAR models discriminated among two categories with accuracies of >96% and the regression QSAR models precisely predicted the mutagenic activity of diverse chemicals yielding high correlations (R ²) between the experimental and model-predicted values in the respective training (>0.96) and test (>0.94) sets. The test set root mean squared error (RMSE) and mean absolute error (MAE) values emphasized the usefulness of the developed models for predicting new compounds. Relevant structural features of diverse chemicals that were responsible and influence the mutagenic activity were identified. The applicability domains of the developed models were defined. The developed models can be used as tools for screening new chemicals for their mutagenicity assessment for regulatory purpose.