A UPLC-MS/MS method for quantification of ß-N-methylamino-L-alanine (BMAA) in Cycas sphaerica roxb. and its use in validating efficacy of a traditional BMAA removal method.

The presence of neurotoxin ß-N-Methylamino-L-alanine (BMAA) in the seeds of Cycas sphaerica is reported for first time. We developed a UPLC-MS/MS method for BMAA quantification by derivatizing with dansyl chloride. The method successfully differentiated L-BMAA from its structural isomer 2,4-diaminobutyric acid (DAB). The extracting mixture 0.1M TCA: ACN 4:1 v/v had a recovery level of >95%. The method is a high throughput sensitive chromatographic technique with 16.42 ng g-1 Limit of Quantification. BMAA was present in the endosperm of C. sphaerica, and was not detected in the leaves and pith. Washing of seeds in running cold water for 48 h reduced BMAA content by 86%. The local communities also treat the seeds under running cold water, but only for 24 h. The results of the study thus validated the traditional BMAA removal process through cold water treatment, but recommend for increase in the treatment period to 48 h or more.

A comprehensive review on spatial and temporal variation of arsenic contamination in Ghaghara basin and its relation to probable incremental life time cancer risk in the local population.

Spatial and temporal variations have been found in the levels of arsenic (As) throughout the groundwater of the Ghaghara basin. Fifteen out of twenty-five districts in this basin are reported to be affected by As, where the levels of As in groundwater and soil exceed the permissible limits set by the WHO (10 µgl-1) and FAO (20 mgkg-1) respectively. These districts include a total of four municipalities in Nepal and eighty-six blocks in India, all of which have varying degrees of As contamination. Approximately 17 million people are at risk of As poisoning, with more than two orders of magnitude higher potential lifetime incremental cancer risk, constituting over 153 thousand potential additional cases of cancer due to As-contaminated drinking water. Out of the 90 As-contaminated blocks in the Ghaghara basin, 4 blocks have about 7-fold higher potential risk of developing cancer, 49 blocks have 8-37-fold higher risk, and 37 blocks have up to 375-fold higher risk compared to the upper limit of the USEPA acceptable range, which is 1 × 10-6-1 × 10-4. High accumulation of As has been reported in the nails, hair, and urine of local inhabitants, with higher levels observed in females than males. The toxicity of As is manifested in terms of a higher occurrence of various diseases. Reproductive endpoints, such as increased incidences of preterm birth, spontaneous abortion, stillbirth, low-birth weight, and neonatal death, have also been reported in the basin. The level of As in tube wells has been found to be negatively correlated with the depth (r = -0.906), and tube wells with high levels of As (>150 µgl-1) are generally located within close proximity (<10 km) to abandoned or present meander channels in the floodplain areas of the Ghaghara river. In addition to As contamination, the water quality index (WQI) in the Ghaghara basin is poor according to the BIS standards for drinking water. Groundwater in six out of fifteen districts is unsuitable for drinking purposes, with a WQI exceeding 100. The levels of As in agricultural soil in many villages of Ballia, Bahraich, and Lakhimpur Kheri districts have exceeded the FAO limit. Water from deep tube wells has been found to be relatively safe in terms of As content, and thus can be recommended for drinking purposes. However, the use of surface water needs to be encouraged for irrigation purposes in order to preserve soil health and reduce As contamination in the food chain, thereby minimizing the risk of cancer.

Changes in fatty acids in Brassica juncea L. oil grown under two simulated conditions of fluoride contamination.

Rapeseed, the second-most-important vegetable oil source, is cultivated in various areas of India where both groundwater and soil are contaminated with fluoride (F-). Furthermore, the frequent use of F- contaminated groundwater for irrigation leads to accumulation of F- in surface and sub-surface soil. The study aims to compare the morphological and biochemical changes in Brassica juncea L., the variations in its fatty acids (FAs) composition and oil yield, under two regimes of F- contaminated soils: (i) pre-contaminated soil (Tr) and (ii) irrigation with F- contaminated water (Ir). The level of F- (µg g-1) in the plant tissues (root, leaf, and grain) was significantly higher in Ir_10 (18.3, 14.7, and 2.8, respectively) than in Tr_10 (4.3, 2.6, and 0.77, respectively), while the oil yield was significantly lower with Ir_10 (19.5%) than with Tr_10 (44.9%). The phytoremediation potential of F- by Brassica juncea L. is greater in Tr regime than in the Ir regime. The erucic acid content (%), which is detrimental to cardiac health, increased to 67.37% (Ir_10) and 58.3% (Tr_10) from 57.73% (control). Thus, the present study shows that irrigation with F- contaminated water results in greater toxicity and accumulation in plants and is not safe for human health.

Irrigation with F^­ contaminated water results in a greater accumulation of F^­ in mustard than cultivated on pre-contaminated soil. The level of erucic acid in mustard oil enhances against F^­ exposure.

Spatio-temporal variation of fluoride in groundwater and agricultural soil and crops of Unnao district, UP: Monitoring and assessment.

Fluoride (F-) contamination in groundwater of Unnao district, Uttar Pradesh was reported for the first time in 1994, however comprehensive monitoring of F- in different environmental matrices remains to be undertaken. The presented study reports spatio-temporal monitoring of F- content in groundwater, crops and soil from F- affected district Unnao, in pre-monsoon (PRM), monsoon (MO) and post-monsoon (PMO), to establish F- groundwater-soil-plant continuum. More than 80% of groundwater samples were contaminated with F-> 1.0 mg L-1 with highest level (mg L-1), at Patiyara (3.6 ± 0.64), during PRM > Pathakpur (2.73 ± 0.57) during PMO > Sarukheda (2.40 ± 0.43) during PRM. High Cr in groundwater was observed in Jajmau (7.08 ± 1.42). The level of F- (mg Kg-1) in agricultural soils followed 3.4 ± 0.71 at Patiyara (MO) > 2.9 ± 0.14 at Badlikheda (PRM) 1.89 ± 0.28 at Jagatkhera (PRM). Among the different edible parts of crops in selected sites, highest F- content (mg Kg-1), F- level in grains of Oryza sativa ranged between 0.23 ± 0.02 to 2.01 ± 0.24. Whereas in the edible fruit of Trichosanthes diocia contained 1.47 ± 0.32 and Momordica charantia 1.47 ± 0.02. Leaf of spinach (1.03 ± 0.22) and seed of Brassica juncea (0.73 ± 0.08). Overall, comparing across all the three seasons, level of F- was highest in all the plants during MO, as compared to PRM and PMO. The regression analysis of physiochemical properties of groundwater show negative relationship between Na+ and F- whereas soil alkalinity exhibited strong influence in soil F-. The high F- content in soil and groundwater at Patiyara and Shekhpur also coincided with presence of several brick kilns, possibly contributing to the high F-.

Nitric oxide-mediated alleviation of arsenic stress involving metalloid detoxification and physiological responses in rice (Oryza sativa L.).

Rice is a staple crop, and food chain contamination of arsenic in rice grain possesses a serious health risk to billions of population. Arsenic stress negatively affects the rice growth, yield and quality of the grains. Nitric oxide (NO) is a major signaling molecule that may trigger various cellular responses in plants. The protective role of NO during arsenite (AsIII) stress and its relationship with plant physiological and metabolic responses is not explored in detail. Exogenous NO, supplemented through the roots in the form of sodium nitroprusside, has been shown to provide protection vis-à-vis AsIII toxicity. The NO-mediated variation in physiological traits such as stomatal density, size, chlorophyll content and photosynthetic rate maintained the growth of the rice plant during AsIII stress. Besides, NO exposure also enhanced the lignin content in the root, decreased total arsenic content and maintained the activities of antioxidant isoenzymes to reduce the ROS level essential for protecting from AsIII mediated oxidative damage in rice plants. Further, NO supplementation enhanced the GSH/GSSG ratio and PC/As molar ratio by modulating PC content to reduce arsenic toxicity. Further, NO-mediated modulation of the level of GA, IAA, SA, JA, amino acids and phenolic metabolites during AsIII stress appears to play a central role to cope up with AsIII toxicity. The study highlighted the role of NO in AsIII stress tolerance involving modulation of metalloid detoxification and physiological pathways in rice plants.

Nickel and cadmium phytoextraction efficiencies of vetiver and lemongrass grown on Ni-Cd battery waste contaminated soil: A comparative study of linear and nonlinear models.

A comparative assessment of the phytoremediation efficiency of two tolerant grass species viz. vetiver and lemongrass were performed in pots against simulated Ni-Cd battery electrolyte waste (EW) contaminated soil (EW1%, EW2% and EW4% w/w). Ni (µg g-1) accumulation was higher in shoots (36.8) and roots (252.9) of vetiver than in lemongrass (12.5 and 79.7, respectively). While the same trend was true for Cd (µg g-1) accumulation in vetiver and lemon grass roots (232.2 and 147.2, respectively), however, the accumulation in vetiver shoot (43.4) was less than in lemongrass (99.9). The bioaccumulation factor of metals in both grasses increased with EW contamination. Vetiver was tolerant towards EW toxicity than lemongrass, as it exhibited lesser decline in morphological parameters, lesser rise in TBARS against the doses of EW. The activities of SOD, APX, POD enzymes were higher in vetiver whereas, only GR in lemongrass. Multiple linear regression model show, pH had strong and positive influence over the Ni and Cd uptake by the plants whereas, phosphate, OM and bioavailable metals influenced negatively. The higher R2 (>0.9) and Chi-square values ≤ 1 in sigmoid non-linear model demonstrates robustness of the model for predicting the Ni and Cd accumulation (MHM) in both the grasses. Ni accumulation was higher than Cd, roots had greater accumulation of heavy metal and vetiver was a greater accumulator of Ni and Cd from EW the contaminated soil than lemongrass.

Biochar amendment reduced the risk associated with metal uptake and improved metabolite content in medicinal herbs.

Contaminations of heavy metals such as lead (Pb) and cadmium (Cd) in medicinal plants (MPs) not only restrict their safe consumption due to health hazards but also lower their productivity. Biochar amendments in the soil are supposed to immobilize the toxic metals, improve the soil quality and agricultural productivity. However, the impact of biochar on growth attributes, metal accumulation, pharmacologically active compounds of MPs, and health risk is less explored. An experiment was performed on three medicinal plants (Bacopa monnieri (L.), Andrographis paniculata (Burmf.) Nees, and Withaniasomnifera (L.)) grown in a greenhouse in soil co-contaminated with Pb and Cd (at two concentrations) without and with biochar amendments (2 and 4% application rates). The fractionation of Pb and Cd, plant growth parameters, stress enzymes, photosynthetic capacity, pharmacologically active compounds, nutrient content, uptake and translocation of metals, antioxidant activities, and metabolite content were examined in the three MPs. The accumulation of Pb and Cd varied from 3.25-228 mg kg1 and 1.29-20.2 mg kg-1 , respectively, in the three MPs, while it was reduced to 0.08-18 mg kg-1 and 0.03-6.05 mg kg-1 upon biochar treatments. Plants grown in Pb and Cd co-contaminated soil had reduced plant biomass (5-50% depending on the species) compared to control and a deleterious effect on photosynthetic attributes and protein content. However, biochar amendments significantly improved plant biomass (21-175%), as well as photosynthesis attributes, chlorophyll, and protein contents. Biochar amendments in Pb and Cd co-contaminated soil significantly reduced the health hazard quotient (HQ) estimated for the consumption of these medicinal herbs grown on metal-rich soil. An enhancement in secondary metabolite content and antioxidant properties was also observed upon biochar treatments. These multiple beneficial effects of biochar supplementation in Pb and Cd co-contaminated soil suggested that a biochar amendment is a sustainable approach for the safe cultivation of MPs. This article is protected by copyright. All rights reserved.

Orthosilicic acid (OSA) reduced grain arsenic accumulation and enhanced yield by modulating the level of trace element, antioxidants, and thiols in rice.

Arsenic (As), a toxic metalloid, is finding its route to human through intake of As-contaminated water and consumption of food grown on contaminated soil. Rice is the most As-affected crop. Present study is aimed to assess the impact of stabilized orthosilicic acid (a proprietary formulation for plant-available silicon (Si) and earlier used as fertilizer for rice to enhance growth and yield) in reducing the accumulation of As in rice grains. Application of arsenic in the form of arsenate (AsV) and arsenite (AsIII) significantly affected plant growth in a dose-dependent manner. Higher doses of AsV and AsIII (50 and 25 mg L-1 respectively) significantly decreased the yield attributes leading to lower yield. A significant accumulation of As in grain was observed in both AsV- and AsIII-exposed plants in a dose-dependent manner. Arsenic exposure also increased the level of Si in rice grains. Application of Si, either in soil or on leaves (foliar), greatly reduced grain As accumulation (up to 67% in AsV and 78% in AsIII) and enhanced the growth and yield of plants under As stress. The level of thiols and activities of antioxidant enzymes were also enhanced under Si application. Foliar Si application was more effective in increasing grain Si level and reducing grain As than soil Si. The level of other trace elements was also significantly enhanced by Si application irrespective of the presence or absence of As in comparison with control. Arsenic exposure constrained some of the trace elements, such as Zn and Co, which were restored by Si application. Results of the present study showed that the application of currently used Si formulation may effectively reduce grain As level even in highly As-contaminated soil and improve grain quality of rice.

Fluoride distribution and contamination in the water, soil and plants continuum and its remedial technologies, an Indian perspective- a review.

Fluorine is an essential element required in trace amounts but gets toxic for human beings at levels more than 1.5 mg F- L-1 primarily through drinking contaminated water. It is the 13th most abundant element and constitutes about 0.06-0.09% in the earth crust. It is electronegative in aqueous medium forming fluoride ion (F-). Fluoride contamination in the environment occurs mostly due to anthropogenic and geogenic sources. Fluoride is widely distributed in all components of environment, air (0.1-0.6 µg L-1) soils (150-400 mg Kg-1) rocks (100-2000 mg Kg-1), plant (0.01-42 mg Kg-1) and water (1.0-38.5 mg L-1). Human beings and animals are being exposed to F- primarily from water (0.2-42.0 mg L-1) and plants (0.77-29.5 µg g-1). Fluorosis, a health hazard due to F- is a major problem in many countries across the world affecting about 200 million people globally. In India, > 62 million people in twenty states are facing problem due to F-. The most affected states are Rajasthan (7670 habitations), Telangana (1,174 habitations) and Karnataka (1122 habitations). To mitigate this problem, there is an urgent need to understand the current status and brief knowledge of F- geochemistry. The objective of this review is to highlight different sources of F- that contaminate different environmental matrices including plants, the extent of contamination level in India, uptake, translocation and toxicity mechanism in plants. The review also highlights currently available mitigation methods or technologies through physio-chemical and biological means.

Comparison of two ferns (Adiantum capillus-veneris Linn. and Microsorium punctatum (Linn.) Copel) for their Cr accumulation potential and antioxidant responses.

Study was undertaken to compare Cr accumulation in two ferns (Adiantum capillus-veneris Linn. and Microsorium punctatum (Linn.) Copel) and the role of antioxidants were also investigated towards metal tolerance in order to assess the use of ferns in phytomediation/ phytostabilization. Different concentrations (0, 50, 100, 150 microg g(-1) dw) of Cr were added to fern planted in pot containing 1 kg soil. In both the ferns, Cr accumulation increased with increase in metal concentration and maximum accumulation of 800.5 microg g(-1) (fronds) and 1457.4 microg g(-1) (roots) in M. punctatum and 660.8 microg g(-1) (fronds) and 1259.6 microg g(-1) (roots) in A. capillus-veneris was recorded. The increase in the levels of malondialdehyde, antioxidants and antioxidant enzymes (superoxide dismutase, glutathione peroxidase) in A. capillus-veneris was less pronounced than M. punctatum under Cr exposure as compared to their respective controls. In view of less decrease in chlorophyll content and antioxidants along with higher accumulation of Cr in the fronds M. punctatum, is indicative of its higher tolerance towards Cr. However, bioaccumulation factor (concentration of Cr in fronds/concentration of Cr in the soil) of both the ferns was recorded > 1 which qualifies the plants as potential Cr hyperaccumulator and suitable for phytoremediaton.

Effect of chromium on accumulation and antioxidants in Cucumis utillissimus L.: response under enhanced bioavailability condition.

This study compares the accumulation of Cr(VI) and biochemical changes (total chlorophyll, carotenoid, protein, malondialdehyde (MDA) and cysteine contents) and roles of antioxidant enzymes (superoxide dismutase (SOD), guaiacol peroxidase (GPX), ascorbate peroxidase (APX)) in tolerance to metal induced stress in Cucumis utillissimus L. grown in Cr contaminated soil (CS) with garden soil (GS). Furthermore, Cr bioavailability was enhanced by ethylene diamine tetra-acetic acid (EDTA) addition to the soil to forecast the plant's accumulation pattern at elevated Cr environment. Accumulation of Cr in the leaves of the plant increased with increase in substrate metals concentration. It further increased with the addition of EDTA by 1437% and 487% in GS and CS, respectively at the highest treatment level. The lipid peroxidation increased proportionately with increase in Cr accumulation in the leaves. All the activity of antioxidant enzymes (SOD, GPX and APX) and the level of cysteine increased with dose dependant manner. SOD and cysteine were observed to be higher in the GS than in CS, but APX and GPX were found to be higher in CS than in GS. The increase in GPX and APX activities with the increase in Cr concentration could be assumed that these two enzymes have a major role in the defense mechanism towards stress induced by Cr in C. utillissimus.

Study on arsenate tolerant and sensitive cultivars of Zea mays L.: differential detoxification mechanism and effect on nutrients status.

The study identifies sensitive and tolerant cultivars of Zea mays L. (cv. Azad kamal (AK) and Azad uttam (AU)) towards As(V) induced stress, based upon growth biochemical parameters and metal(loid) levels in a sand culture. As(V) (µgg⁻¹ dw) accumulation was lower in cv. AK (31 ± 1 and 107 ± 30) than cv. AU (34.5 ± 3.3 and 132.6) in leaves and roots, respectively, which correlated with lower levels of malondialdehyde and H2O2. No definite trend of Mn, Cu, Zn, Fe, Ca, K and Na accumulation signifies that As(V) has little influence on their uptake. Total chlorophyll and protein levels increased in cv. AK and decreased in cv. AU at 7d. Higher levels of SOD and GR in cv. AK and conversely higher levels of APX, GPX and CAT in cv. AU could be a possible differential detoxification mechanism between the cultivars. The results indicate that cv. AK seems to be arsenate tolerant than cv. AU. We assure that the undertaken study does not involve humans or experimental animals and were conducted in accordance with national and institutional guidelines for the protection of human subjects and animal welfare.

Metal accumulation, growth, antioxidants and oil yield of Brassica juncea L. exposed to different metals.

In agricultural fields, heavy metal contamination is responsible for limiting the crop productivity and quality. This study reports that the plants of Brassica juncea L. cv. Pusa bold grown on contaminated substrates [Cu, Cr(VI), As(III), As(V)] under simulated field conditions have shown translocation of metals to the upper part and its sequestration in the leaves without significantly affecting on oil yield, except for Cr and higher concentration of As(V), compared to control. Decrease in the oil content in As(V) treated plants was observed in a dose dependent manner; however, maximum decrease was recorded in Cr treated plants. Among all the metal treatments, Cr was the most toxic as evident from the decrease in oil content, growth parameters and antioxidants. The accumulation of metals was below the detection limit in the seeds grown on 10 and 30 mg kg(-1) As(III) and Cr(VI); 10 mg kg(-1) As(V)) and thus can be recommended only for oil cultivation.

Interactive effects of Cr and Fe treatments on plants growth, nutrition and oxidative status in Zea mays L.

The effects of Cr and Fe, singly and in combination were investigated on nutrients uptake (Cu, Zn and Mn), lipid peroxidation, antioxidant enzymes, chlorophyll content and growth parameters in Zea mays L. var. Ruchi (SRHM 445). Roots of the Cr treated plants were stunted and root hair formation was greatly impaired. The leaves were bearing wilted appearance. Cr was primarily accumulated in the roots with a low translocation rate to the leaves. Cu absorption decreased with increase in Cr concentrations only in roots. In leaves there is no correlation. Iron induced lipid peroxidation was higher in the leaves at both the concentrations (3 and 9 microg ml(-1)) after 7d, and decreased significantly by the addition of Cr. Concentrations of most leaves antioxidants were lower in mixed metal treatment compared to single treatments, indicating an interaction between metals leading to reduced cellular effects as indicated by lower lipid peroxidation levels. Changes in APX and GPX activities observed in the leaves of contaminated-plants suggest their involvement in heavy-metal stress tolerance. The plant seems to be tolerant as the translocation of Cr was recorded less, which decreased in the presence of Fe.

Role of antioxidants in Cr tolerance of three crop plants: Metal accumulation in seeds.

Pot experiments were conducted on three crop plants (Vigna radiata cv. PDM54, V. radiata cv. NM1, Brassica juncea cv. vaibhav) which were grown on six different tannery sludge (TS) amendments. The translocation of Cr to the seeds was found to be higher in B. juncea than the two cultivars of V. radiata, and their levels were below detectable limits in all the three plants grown on lower amendments of tannery sludge (T10 and T25). There was a gradual increase in protein content and antioxidant levels in all the plants grown on lower sludge amendments. However, the levels of all the antioxidants were higher in B. juncea than the two cultivars of V. radiata. The increase in malondialdehyde content of B. juncea was lower than the two cultivars of V. radiata as compared to their respective controls. Overall, the plants of B. juncea have shown better tolerance than both the cultivars of V. radiata.