DELINEATION OF CADMIUM CONTAMINATED SOILS AROUND BUDDAH NALLAH (LUDHIANA) AND REMEDIAL MEASURES OF AFFECTED SOILS

Thesis

Submitted to Punjab Agricultural University in partial fulfillment of the requirements for the degree of

MASTER OF SCIENCE

in

SOILS
(Minor Subject: Agronomy)

By
Dharamvir Singh Kambo

(L-2008-A-77-M)

Department of Soil Science

College of Agriculture

PUNJAB AGRICULTURAL UNIVERSITY LUDHIANA-141004

2011
1

CERTIFICATE I
This is to certify that the thesis entitled, Delineation of cadmium contaminated soils around Buddah Nallah (Ludhiana) and remedial measures of affected soils submitted for the degree of M.Sc. in the subject of Soil Science (Minor Subject: Agronomy) of the Punjab Agricultural University, Ludhiana, is a bonafide research work carried out by Dharamvir Singh Kambo (L-2008-A-77-M) under my supervision and that no part of this thesis has been submitted for any other degree. The assistance and help received during the course of investigations have been fully acknowledged.

(Major Advisor) Dr. MPS. KHURANA Senior Soil Chemist Department of Soil Science Punjab Agricultural University, Ludhiana- 141004 (India)

CERTIFICATE- II 2

This is to certify that the thesis entitled, Delineation of cadmium contaminated soils around Buddah Nallah (Ludhiana) and remedial measures of affected soils submitted by Dharamvir Singh Kambo (L-2008-A-77-M) to the Punjab Agricultural University, Ludhiana, in partial fulfilment of the requirements for the degree of M. Sc. in the subject of Soil Science (Minor Subject: Agronomy ) has been approved by the Students Advisory Committee along with the Head of Department after an oral examination of the same.

___________ Head of the Department (Dr. Charanjit Singh) Major Advisor (Dr.MPS Khurana)

_________________________ Dean, Post-Graduate Studies (Dr. Gursharan Singh)

Acknowledgements
First of all, I bow my head to AKAL PURKH the ALMIGHTY by whose kindness I have been able to clear another chapter of my life. Words are compendious in expressing my profound sense of gratitude to my revered Major Advisor Dr. M P S Khurana, Sr. Soil Chemist, Department of Soil Science, Punjab Agricultural University, Ludhiana, for his constant guidance, constructive criticism, encouragement and unstinting moral support provided during this investigation. Working under his expertise has been a great learning experience. I am highly thankful to the respected members of my Advisory Committee, Dr.U S Sadana, Professor of Soil fertility, Department of Soil Science, Dr. B S Brar, Sr. Soil Scientist Department of Soil Science and Dr. Raj Kumar Pedologist, Department of Soil Science and Dr. S S Walia Agronomist, Department of Agronomy for their valuable suggestions, continuous support and going through the manuscript. I duly acknowledge the research facilities provided by the Head, Department of Soil Science, Punjab Agricultural University, Ludhiana. I am falling short of words to express my feelings of obligation toward my parents who always stand by me during the testing times of my life. Their everlasting inspiration, support and affection enable me to face difficult situations in my life. I am highly thankful to my senior Mr. Sanjeev Kumar and my friends, Jasbir, Arshdeep, Sukhwinder, Satnam, Gurpreet, Sandeep and Remander, for their pleasant association and cooperation during the hours of need. Invaluable help rendered by laboratory and field staff of the Department of Soil Science is fully acknowledged. I feel proud to be a part of PAU, Ludhiana where I learnt a lot and spent some unforgettable moments of my life. Needless to say, errors and omissions if any are all mine.

Date__________ ______________ Place: Ludhiana Singh Kambo

Dharamvir

Title of the thesis Name of the student and Admission No. Major Subject Minor Subject Name and Designation of Major Advisor Degree to be Awarded Year of award of degree Total pages in thesis Name of the University

: : : : : : : : :

Delineation of cadmium contaminated soils around Buddah Nallah (Ludhiana) and remedial measures of affected soils Dharamvir Singh Kambo L-2008-A-77-M Soil Science Agronomy Dr. MPS Khurana, Sn. Soil Chemist M.Sc. 2011 80 Punjab Agricultural University, Ludhiana 141 004, Punjab, India ABSTRACT

Cadmium is potentially toxic metal and is highly carcinogenic that enters the food chain from the soil through crop uptake resulting from various anthropogenic activities. The surface sewage irrigated soils collected laterally around Buddah Nullah irrespective of the sites had DTPA extractable cadmium 5.2 times more than the adjoining tube well irrigated soils. Considering the threshold value of 3 mg Cd kg-1 soil, about 11.3 per cent soils have crossed this limit and needs cleanup operation. However management option to rehabilitate such soils depends on pools of Cd responsible for phytotoxicity and use of amendments able to influence these pools. A screen house experiment was conducted to assess the effect of (0, 2.5, 5 10, 20 and 40 mg Cd kg-1 soil) and CaCO3 (2.5 and 5%), FYM (1 and 2 %) and Phosphorous (20 and 40 P2O5 mg kg-1 soil) on the growth of pigweed on soil having DTPA-Cd 0.36 mg kg-1 soil. Dry matter yields decreased as a consequence of phytotoxic effect of Cd emanating from increased availability of Cd in soils and plants. The rate at which significant declined occurred was 10 mg kg-1. However application of different amendments viz (Calcium carbonate, FYM, Phosphorus) exhibited variable behavior as far as their remediation potential was concerned. Application CaCO3, FYM and phosphorous at their highest rate reduced DTPA- Cd by 52.6 percent, 37.1 percent and 45.1 percent respectively. Consequently maximum enhancement in dry matter yields was observed with application of 5% CaCO3 among other amendments. The upper critical toxic level in soil and shoots of pig weed was found to be 4.38 mg kg -1 soil and 14.6 g g-1 dry matter respectively. The interaction of Cd with Zn and Fe was found to be rate dependent. Cu and Mn in both shoots and roots was negatively correlated to the added Cd. All the fractions of Cd in soils increased significantly with cadmium application. Amendments decreased the EX+WS fraction, the maximum depressing effect observed with 5% CaCO3 because of concomitant increase in CARB fraction. FYM application decreased the CARB fraction where as it encouraged both OM-Cd and oxide bound fractions. Phosphorus application was effective in transforming Cd in to oxide bound fractions with little influence on carbonate and organic. Key words: Cadmium, Delineation, Amendments, Critical levels, Cadmium fractions

________________________

___________________

Signature of the Major Advisor

Signature of the Student

CONTENTS CHAPTER I II III IV V TOPIC INTRODUCTION REVIEW OF LITERATURE MATERIALS AND METHODS RESULTS AND DISCUSSION SUMMARY REFERENCES PAGE

1-3 4-21 22-27 28-62 63-66 67-80

CHAPTER - I INTRODUCTION Increasing public awareness over the effects of environmental pollution during recent decade was a result of enhanced understanding of the risk to human health. Contamination of elevated-heavy metals in soils can adversely affect soil ecology, agricultural productivity, quality of agricultural products, water resources, human and animal serious health problem (Raicevic 2005). Agricultural lands show elevated levels of pollutant elements due to various anthropogenic activities such as continuous use of sewage water largely contaminated with industrial effluents, sewage sludge and fertilizers (Rattan et al 2002; Patel et al 2004). It is released into environment by power stations, industries engaged in electroplating, pigments, plastics, stabilizers and nickel cadmium batteries (Sanita di Toppi and Gabrielli 1999). Increasing awareness of the environmental and public health hazard of toxic metals pressurizes society to develop management strategies to remediate or restore the contaminated area. Among the pollutant elements, cadmium (Cd) is of great environmental concern as it is easily absorbed by the plants from the contaminated soils and frequently accumulated by agriculturally important crops with a significant potential to impair animal and human health. Cadmium is a pollutant and potential toxin that has no known function in any biological organism (Wagner 1993). This warrants close attention as it is a cumulative poison. This has led the International Food Standards Organization, Codex Alimentarious Commission to propose a 0.1 mg Cd kg-1 limit for cereals, pulses and legumes. The maximum tolerable intake of cadmium for humans recommended by FAO/WHO is 70 ug/day. Therefore demarcation, delineation and mapping through global poisoning system of cadmium contaminated soil in the highly industrialized town of Punjab (Ludhiana) is urgently required to know the extent of pollution. Among remediation options, physical methods such as vitrification and evacuation of polluted soils and its disposal to land fill sites are quite expensive. The major problem associated with phyto-remediation is low metal removing rates (McGrath et al 2002). Chemical stabilization appears to be an alternative technique that is seen as cost-effective and environmentally sustainable. The principal aim of stabilization is the reduction in the bio7

available fraction of the metal either through increased metal sorption and/or precipitation, or through the formation of insoluble complexes. Hence; there has been an increasing interest in the immobilization of the metals using range of inorganic and organic material like lime, phosphate, organic manures, and zeolites (Singh et al 1989; Khurana and Kansal 2000). However, use of lime, phosphate, and farm yard manure (FYM) is cost-effective option. Lime provides adsorptive surfaces and facilitates pH induced precipitation. The FYM as metal ameliorant has also been studied as it off sets the toxic effect of pollutant elements. On the contrary, Narwal and Singh (1998) reported an increase insolubility of metals in soils amended with organic material. Further speciation in to various geochemical forms of heavy metals including cadmium in contaminated soils affecting their solubility is gaining importance to understand fully the behavior of this metal in soils which directly influence their availability to plants. Although the total metal concentration in many contaminated soils may be high, the phytoavailable fraction is usually very low due to the strong association of metal with organic matter, FeMn oxides, and clays, and precipitation as carbonates, hydroxides, and phosphates (McBride 1995). The magnitude of these forms is not only controlled by pH, organic matter, cation exchange capacity, calcium carbonate content of the soil through dissolution, adsorption, precipitation and chelation reaction but also on the rates of metal loading from inorganic or organic sources and climate. Once, these heavy metals are incorporated in to the soil, their extractability decreases with time indicating a possible change of their forms in the soil (Bell et al 1991). If the crop is planted following the application of heavy metals and amendments (Calcium carbonate, FYM and Phosphorus), the change in chemical form any influence the uptake by plants and help to reduce the toxicity. Effect of different inorganic amendments on fractionation and availability of Cd to wheat was evaluated by Ghafoor et al (2008). Inorganic amendments viz. lime, gypsum, diammonium phosphate (DAP) and potassium dihydrogen phosphate was used at different rates were found to effective in retaining Cd in non available pools and substantial decrease in exchangeable fraction of Cadmium. Sequential extraction data thus indicated that amendments were equally effective in transforming readily available Cd to less mobile fractions (carbonate, Fe/Mn oxide). There is paucity of information pertaining to speciation (various fractions) controlling and contributing towards the availability of cadmium to the crops particularly grown on sewage irrigated soils. This study was to examine the effect of amendments on fractionation and bioavailability of Cd in soil in order to the alleviate its toxic effects of cadmium on vegetables crops. In Punjab, vegetables crops are grown mostly on sewage irrigated soils and are known to accumulate large amounts of heavy metals in their shoots and roots because of their high biomass and root proliferation. Chalai (Pig weed Amaranthus tricolor) is generally 8

found growing on sewage irrigated soils for edible purposes because of the low production cost and high productivity per unit area, this is considered to be the low cost vegetable in the Indian market and often described as poor mans food. As it is consumed as green vegetable, amount of Cd present in it has direct bearing on human health. Although various amendments has been tried for different crops but the work on efficient and cost efficient ameliorative measure affecting the availability of cadmium to this crop is still not elucidated. There for it was felt desirable to examine different amendments on the bioavailability of Cd to this crop. Objectives 1) 2) 3) 4) To delineate Cd contaminated soil for its lateral distribution in order to assess its extent of pollution. To assess relative suitability of different amendments for minimizing Cd pollution in contaminated soils and plants. To establish the upper threshold limit of toxicity of Cd for pig weed. To study the effect of various amendments on the transformations of Cd in soil.

CHAPTER II
REVIEW OF LITERATURE The review pertinent to present investigation had been reviewed under the following heads 2.1 2.2 2.3 2.4 Influence of sewage water / sludge application on accumulation of heavy metal in soils and plants Effect of cadmium on growth, concentration and uptake in different plant species: Effect of different amendments on cadmium uptake Transformation of heavy metals in soils

2.1 Influence of sewage water / sludge application on accumulation of heavy metal in soils and plants Repeated use of waste water for irrigation to crops resulted in build-up of heavymetals in the soil (Singh et al, 1985; Mapanda et al 2005; Rattan et al. 2005; Liu et al. 2005; Mitra and Gupta 1999; Zhang et al 2008). Elevated levels of different heavy-metals in soils regularly irrigated with sewage water than ground water have also been found (Siebe 1998; Aghabarati et al. 2008). Siebe (1998) reported significant increase in DTPA-extractable-Pb and Cd up to 30 cm soil layer with the application of sewage water, consecutively for 80 years than the soils irrigated with ground water. El-Arby and Elbordiny (2006) reported concentrations of DTPA-Fe, Mn, Zn, Cu, Co and Ni in the sandy soil receiving treated waste water averaged 3.2, 122, 129, and 186 which was 22.0, 14.5 and 10.5 fold than soils receiving ground water as irrigation, respectively. Mapanda et al (2005) studied the magnitude of contamination and annual loadings of soils with Cu, Zn, Cd, Ni, Cr and Pb at three sites in Harare (Zimbabwe) where waste water irrigation was practiced in vegetable gardens for last ten years. They reported that heavy-metal concentrations in sandy and sandyclay soils ranged from 7.0 to 145 mg kg1 for Cu, 14 to 228 mg kg1 for Zn, 0.5 to 3.4 mg kg1 for Cd, <0.01 to 21 mg kg1 for Ni, 33 to 225 mg kg1 for Cr and 4 to 59 mg kg1 for Pb up to 20 cm soil depth. Annual heavy metal loading rates showed that all studied heavy-metals exceeded their permitted limits in soils, depending on site. Mitra and Gupta (1999) reported that total and DTPAextractable concentration of Pb, Cd, Cr, Co and Ni were significantly higher in soils of East Calcutta irrigated with sewage water than the tube well water irrigated soils of Baruipur farm. Concentration of Pb, Cd and Cr were found to be above the permissible limits in sewage irrigated soils. 10

Singh et al (2010) reported that continuous application of waste water and clean water site as have led to higher concentrations of heavy metals in the soil at waste water irrigated site as compared to clean water irrigated site which were higher by 109 % for Cd, 152 % for Cu, 25 % for Pb, 32 % for Zn, 161 % for Ni and 52.8 % for Cr. Kansal and Khurana (1999) observed that DTPAextractable and total Cd concentration of sewage irrigated soils were higher than tube-well irrigated soils in the industrial towns of Punjab. Krsad Trkdo et al (2003) reported 2 to 50-fold higher concentration of Cd, Pb, Cu and Co, ~40-fold higher Zn concentration in volcanic soils irrigated with waste water compared to ground water in Van region of Turkey. Liu et al (2005) reported metal enrichment factor of Cd (1.8), Cr (1.7), Cu (2.3), Zn (2.0), Pb (1.9) and the metal contamination factor of Cd (2.6), Cr (1.5), Cu (2.0), Zn (1.7), and Pb (1.6) in soil that received the sewage water irrigation. Rattan et al (2005) reported that sewage waste water irrigation for 20 years resulted in significant build-up of DTPA-extractable Zn (208%), Cu (170%), Fe (170%), Ni (63%) and Pb (29%) in sewage waste water irrigated soils over ground water irrigated soils, whereas Mn content was depleted by 31%. Fractionation study indicated relatively higher build-up of Zn, Cu, Fe and Mn in bio-available pools of sewage waste water irrigated soils. In contrast, Rusan et al (2007) reported non-significant (p0.05) differences in soil available Pb and Cd content in soils of Jordan with the application of municipal waste water even up to 10 years. Mohammad and Mazahrez (2003) also reported inconsistent variation in heavy-metal content of soil with industrial waste water application. Khurana et al (2003) found that mean values of DTPA- Pb, Ni, Cd Zn, Mn and Fe in surface (0-15cm) soils of highly industrialized city of Ludhiana irrigated largely with sewage effluents were 4.21, 3.58, 0.30, 11.9, 25.4 and 49.2 as compared to 2.76, 0.40, 0.12, 2.10, 8.34, 10.88 in less industrialized city of Sangrur indicating higher loading of soils of Ludhiana with heavy metals through sewage irrigation. The increase in heavy metals content of soils with continuous application of sludge/ sewage water has also been reported by Brar et al (2002) and Kuhad et al (1989). Taywade and Prasad (2008) observed that in sewage water irrigated soils are associated with relatively higher concentration of DTPA-Fe, Mn, Cu, Zn, Pb, Cr and Cd as compared to the corresponding non-irrigated soils. In a study Dheri et al (2007) reported that Pb, Cr, Cd and Ni were significantly higher in sewage water irrigated soil compared to tube well irrigated soil .The concentration of Pb, Cr, Cd and Ni in sewage irrigated soils were 1.8, 35.5, 3.6 and 14.3 times higher than their concentration in tube well irrigated soils respectively. The build up of Cr (35.5 times) and Ni

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(14.3 times) in this study was so high that these elements may cause phyto-toxicity to crop plants with continuous application of sewage effluents. The investigation carried by Sikka (2003) found that the mean content of available cadmium, lead and nickel where Buddah Nallah (a sewage channel passing through the interior of Ludhiana City) water has been continuously used were 0.8, 4.7 and 1.9 mg kg -1 soil which were 530, 361 and 296 percent higher than the adjoining fields where tube well water has been used for irrigation. In earlier study Sharma and Kansal (1986) noted that available cadmium was three to five times higher in Nullah irrigated soils where it ranged from 0.119 to 0.253 mg kg
-1

soil as against its content which ranged from 0.085 to 0.115 mg kg

-1

soil in

tube-well irrigated Gupta and Mitra (2002) studied the effect of long-term use of sewage waste water in soils of Calcutta (India) and reported 2.43, 46.5, 3.81, 0.86, 93.0, 15.9, 3.88, 2.44 and 6.61 fold increase in total-Fe, Zn, Cu, Mn, Cd, Pb, Co, Ni and Cr concentration in comparison to soils receiving ground water for crops. In an another study in El-Sadat City of Egypt on the use of industrial for 5 years, El-Arby and Elbordiny (2006) reported significant increase of 1.72, 2.48, 2.72, 4.49, 4.24, 1.90 and 2.36 times increased concentration of total-Fe, Mn, Zn, Cu, Co, Ni and Pb over ground water irrigated soils (0-15 cm) has been reported. Patel et al (2004) compared the build-up of total heavy-metal content in soils irrigated with mixed sewage and industrial effluents and those irrigated with effluents from paper mill industry. They reported that soil irrigated with paper mill effluents accumulate 2.4, 4.5 and 3.8 mg kg-1 less Fe, Zn and Cu, respectively but 2.1 and 18.6 mg kg-1 more Mn and Pb, respectively than mixed sewage and industrial effluents. Chen et al (2009) analyzed the total concentrations and chemical speciation of heavy metals including Cd, Cr, Cu, Zn and Ni in sewageirrigated soils in the eastern suburb of Beijing, China. The results showed that there was remarkable buildup of Cd, Cr, Zn and Cu in sewage-irrigated top soils compared to reference topsoils. Among the four metals, Cd was more mobile and bioavailable in the sewage-irrigated topsoils than in the reference topsoils. Higher Cd contents in sewage-irrigated soils may constitute potential risk on food security and human health. Prabu et al (2009) conducted an experiment to assess the extent of heavy metal contamination of vegetables due to irrigation with polluted Akaki river water, Ethiopia on agricultural land. The results showed that the heavy metals in Akaki water were higher than the natural elemental levels in freshwater. The concentration of Cr in all vegetables was more than the maximum limit. The Cd accumulation was more in leafy vegetables than other vegetables under study. Metal transfer factors from soil to vegetables were significant for Zn, Mn, Cu, Fe and Cd and accumulation of Cr and Ni was comparatively less while that of Zn, Fe, Cu and Mn was more in vegetable plants. 12

Mapanda et al (2005) studied the contamination of leafy vegetables (Brassica species) with copper (Cu), zinc (Zn), cadmium (Cd), nickel (Ni), lead (Pb) and chromium (Cr) and their subsequent risk to human exposure in the City of Harare, where waste water was used for irrigating vegetables and concluded that it was not safe and might not be sustainable in the long-term. They also reported elevated concentrations of Cu, Zn, Cd, Ni, Cr and Pb in the topsoil of sites irrigated with waste water. Lokeshwari and Chandrappa (2006) studied to assess the extent of heavy metal contamination of vegetation due to irrigation with sewage-fed lake water on agricultural land. Samples of water, soil and crop plants have been analyzed for seven heavy metals, viz. Fe, Zn, Cu, Ni, Cr, Pb and Cd using atomic absorption spectrophotometry. The results showed the presence of some of the heavy metals in rice and vegetables, beyond the limits of Indian standards. Metal transfer factors from soil to vegetation are found significant for Zn, Cu, Pb and Cd. Comparing the results of heavy metals in water, soil and vegetation with their respective natural levels; it was observed that impact of lake water on vegetation was found to be more than the soil. Gupta et al (2007) investigated the effects of municipal waste water irrigation on the accumulation of heavy metals ( Pb, Zn, Cd, Cr, Cu and Ni ) in soil and vegetables and revealed that heavy metal-contaminated vegetables grown in wastewater-irrigated areas may pose public health hazards. Mishra and Tripathi (2008) conducted a study to determine the heavy metal contamination in soil with accumulation in plants in waste water irrigated areas. Results revealed that waste water contained lower concentrations of Cr, Zn, Cu, and Pb except Cd than the permissible limits prescribed by the World Health Organization (WHO). The maximum metal concentrations occurred in Brassica oleracea. The metal enrichment and degree of contamination showed that accumulation of the five toxic metals increased during sewage irrigation as compared with the reference values, other Indian regions and globally. However, based on WHO standards for heavy metal contamination of soil and irrigation water, our data does not ensure safe levels for food. 2.2 Effect of Cadmium on growth, its concentration and uptake in different plant species Cadmium is a non essential element that negatively affects plant growth and development. It is recognized as an extremely significant pollutant due to its high toxicity and large solubility in water (Pinto et al 2004). It is easily taken up by plants and can interfere with many physiological processes associated with normal growth and development (Artexe et al 2002). It inhibits root and shoot growth, affects nutrient uptake and synthesis of chlorophyll, protein, carbohydrate, free amino acids, and RNA etc Greger et al (1991) and reduces crop yields (Jing and Logan 1992; Lune Van and Zwart 1997). The uptake of cadmium by higher plants depends on its concentration in the soil and its bioavailability, 13

modulated by the presence of organic matter, pH, redox potential, temperature and concentration of other elements. Cadmium easily penetrates the root through the cortical tissue and is translocated to the above ground tissues (Yang et al 1998). In general, cadmium is toxic to a number of plant species at relatively low soil and plant concentration.Visible symptoms of cadmium toxicity in plants include chlorosis, leaf rolls and stunting. Patel et al (1980) reported that high level of Cd produced 50% or more yield decrease in bush bean plants (Phaseolus vulgaris) in highly calcareous desert soil. Further, they reported that increasing Cd levels caused significant decrease in the yield of crop. Borges and Wollum (1981) reported that application of Cd decreased the dry matter production of tops, roots and nodules of soybean plants. Nutrient imbalance in the presence of Cd was observed. They suggested that the interaction of Cd with some nutrients may be responsible for Cd toxicity. They further found most pronounced Cd effect on Fe and Mn rather than Zn. Page (1981) found that Cd was usually lowest in grain and fruit crops, intermediate in root crops and highest in leafy crops. The most important soil factors influencing crop Cd were the concentration in soil and soil pH. Cataldo et al (1981) also reported that following root absorption, Cd was strongly retained by roots with only 2% of the accumulated Cd being transported to leaves, as much as 8% was transported to seeds during seed filling. Khan and Khan (1983) studied the effect of Cd on the nutrient concentration of tomato and egg plant. The application of Cd was found to effect the nutrient concentration and their uptake by plants. Cadmium decreased Zn in both crops and increased Mn, Fe and Cu in tomato and decreased in egg plant. Chernykh (1991) noticed the effect of different concentrations of Pb and Cd on the uptake of major and trace elements by barley and Vicia sativa cv which were grown in pots taking various types of soils. The low concentrations of Cd or Pb didnt alter plant material uptake, but higher heavy metal concentrations disrupted mineral uptake and translocation. Reductions in the uptake of P, Ca, Mg, Cu and Fe were more pronounced in Vicia sativa than in barley at high heavy metal concentration. Among the various soils, the effect of Pb and Cd were more visible in acid sod-podzolic soil than in uncultivated sod-podzolic and least in chernozem. Haghiri (1973) studied the growth and cadmium concentration of soybean and wheat tops as influenced by soil applied Cd. In both the crops, Cd concentration increased while yields decreased with increasing level of applied Cd. Cadmium toxicity (2.5 ppm) of applied cadmium. He also assessed the effect of soil applied Cd at 0-10 ppm on the yield and cadmium uptake of radish, lettuce, capsicum and celery. Cadmium tolerance and uptake varied widely among crops, lettuce having the highest concentration and radish roots the lowest. The largest relative reduction in growth was shown by lettuce followed by radish, 14

green pepper and celery. Maclean (1976) found that addition of 5 ppm of Cd decreased the yield of lettuce in all the five soils selected during the study. Rana and Kansal (1985) reported that application of Cd to soil showed adverse effect on plant growth. The fresh yield of barseem fodder decreased significantly with Cd application to soils. The higher rate of Cd application even lead to mortality of plants immediately after germination. Koreak and Fanning (1978) added Cd to soil (2 mg Cd kg-1 soil) in the form of inorganic salt (CdSO4) and an equivalent amount in the form of municipal sewage sludge and determined the amount of Cd absorbed by corn (Zea mays). Even though, the pH of the soil amended with sewage sludge was lower (pH 6.0) than that of same soil amended with CdSO4 (pH 6.5), the amount of cadmium accumulated by the corn foliage from CdSO4 was 5 to 18 times greater than the amount accumulated by the foliage from the soil amended with sewage sludge. Maclean (1976) observed reduction in yield of lettuce in all the five soils when Cd was added at the rate of 5 ppm. Sadana and Singh (1987) found that increasing level of Cd up to 8.0 mg kg-1 soil reduced significantly (40 percent) dry matter yield of spinach. The corresponding increase in Cd content of spinach was from 0.6 to 38.4 g g -1 dry matter. Singh and Nayyar (1991) observed 15 to 79 percent reduction in dry matter yield of wheat at 45 days of growth at different rates of Cd application. Saini and Kansal (1990) revealed that yield of maize fodder decreased significantly with the application of cadmium. The higher yield of 9.24 g pot-1 was obtained without Cd which was significantly reduced to 7.30 g pot -1 when the rate of Cd application was increased to 5 g g-1 soil. Singh and Nayyar (1989) reported a significant decrease in mean dry matter yield of corn with 30 mg Cd kg-1 soil application in a pot culture experiment on coarse textured soil. Juwarkar and Shende (1986) obtained 32 percent reduction in grain yield of wheat on calcareous vertisol due to toxic effect of high level of Cd (400 ppm) applied through sewage sludge. Walker et al (1979) reported a significant linear effect of Cd upon the concentration of Zn, Mn, Cu, K, Ca and Mg except P which decreased with increased Cd conducted on low pH soil application in soyabean plants while Mahler et al (1982) from a green house trial reported that Cd additions to the soil decreased Zn in lettuce, tomato, corn and swiss chard, decreased Mn in Swiss chard and lettuce, it increased in tomato but showed little or no effect in corn. Further, application of Cd didnt affect P, Fe, Ca, Mg, S and K uptake. Jeng and Bergseth (1992) reported that high total content of heavy metals is not synonymous with increased plant uptake. Jasiewicz (1993) sampled carrots, beetroots, parsley, celery, leeks, onions, cabbages and lettuces taken from numerous localities and analyzed for Cd, Ni and Pb levels in the aerial parts as well as in roots. In the majority of

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samples, Cd and Ni accumulated more in the roots as compared to Pb which was more in the aerial parts. The contents of Cd and Pb exceeded the permitted levels. In a green house experiment, Lehoczay et al (1996) studied the influence of Cd concentrations (0, 50 or 100 mg kg-1 soil) on biomass production and Cd contents of maize, garlic and spinach by taking two soil types i.e. Eutric cambisol and Gleycluvisol. The Cd contents of all three crops increased with increasing concentrations of Cd in the soil. The concentrations of Cd in plants grown in the strongly acidic gleycluvisol soil were many times higher than those of plants grown in the neutral Eutric cambisol soil. With regard to biomass accumulation spinach was most sensitive to Cd. Arauja et al (1997) reported that Cd contents in plants like Carinata and Duronegro increased with increased cadmium levels in the nutrient solutions. Applied cadmium decreased contents of Zn and Mn in roots with little or no effect on the contents of these elements in the shoot. Copper content in plants remained unaffected by Cd levels in the nutrient solution except for the Cd content in roots of Carinata, Fe contant was altered only in the roots of duronegro. Bipasha et al (1997) reported that Cd alone significantly decreased shoot bud formation and shoot elongation, with the higher concentration having greater effects, but addition of equal concentration of Zn overcame the toxicity of Cd to some extent. Tolerant plants could be regenerated in a culture medium containing both metals and showed greater uptake of Cd in the roots than shoots, whereas Zn was translocated from roots to shoots. Ebbs and Kochain (1998) screened twenty two grasses and cereals in hydroponic culture indicated that oats (Avena sativa) and barley (Hordeum vulgare) tolerated higher levels of Cu, Cd and Zn as evidenced by elevated concentrations of these metals in the plant shoots. Khurana et al (2006) reported significant decrease in mean dry matter yield of maize cultivars namely Sartaj and Parkash with 10 mg Cd kg-1 soil. Lehoczky et al (1998) assessed the effect of increasing Cd concentration on biomass production of lettuce and noticed a decrease in biomass production as Cd rate was increased. Similar results were obtained by Singh and Aggarwal (2005). A study was conducted by Shafiq (2008) to determine the effect of different concentrations of lead and cadmium on seed germination and seedling growth of Leucaena leucocephala. Seed were grown under laboratory conditions at 25, 50, 75 and 100 ppm of metal ions of lead and cadmium. Both lead and cadmium treatments showed toxic effects on various growth indices of L. leucocephala. Seed germination and root length significantly (p<0.05) decreased at 50 ppm treatment of cadmium as compared to control. The seedling dry weight also significantly (p<0.05) reduced at 25 ppm treatment of lead and cadmium. 2.3 Effect of different amendments on cadmium concentration and its uptake

16

Stabilization/solidification techniques are developed to convert contaminants into less soluble, immobile or less toxic forms. Stabilization aims to immobilize contaminants by adding immobilizing agents enhancing adsorption, complex binding or precipitation (Kumpiene et al 2008). Typical immobilizing agents in field studies amendments are: limes, phosphates, organic matter induced additives (peat, manure) and industrial co-products based Synthetics (Guo et al 2006) 2.3.1 Effect of phosphorus amendment on cadmium concentration and its uptake The application of different amendments is well known for their effectiveness to reduce toxicity of heavy metal to crop plant in contaminated soils. The use of phosphorus amendment has gained an increasing interest to minimize heavy metal toxicity through chemical immobilization of Cd in crop plant. The reduction in phyto toxicity of heavy metal has been observed due to their immobilization in soil. The role of phosphorus amendments to reduce Cd toxicity occurred through the formation of Cdphosphate complexes. This may be due to sorption mechanisms such as surface fixation, ion exchange, precipitation and co-perception when different phosphorus source are added to Cd contaminated soils. These processes led to lowering the phyto availability of Cd to crop plants. Chemical immobilization of heavy metal with phosphate may thus be a good remedial measure to reduce Cd toxicity to crop plants in contaminated sites. Phosphate-based additives forms secondary phosphate precipitates that are relatively insoluble and stable in a wide range of conditions. Solubility of both metals and phosphates remains low under neutral conditions and acidity has to be added for an efficient metal immobilization (Chen et al 2003). Levi-Minzi and Petruzzelli (1984) observed that phosphate induce variation in soil pH influenced the solubility of Cd in soils. They noticed that that while the effect phosphorus on pH and cadmium was less evident in an organic soils with high pH buffering capacity, the addition of diammonium phosphate increase soil ph thereby reducing solubility of Cd in mineral soils with low pH buffering capacity. Bolland et al (1977) reported that the specific adsorption of phosphate anions increased the negative charge of variable surface which may result in the increasing retention of metal cat ion such as Cd +2, Cu+2 and Zn+2. Naidu et al (1996) also reported an increase in net negative charge of surface due to specific adsorption of phosphate anions. Bolan et al (1999) observed that retention of Cd+2, Cu+2and Zn+2 ions increased with increase in specific adsorption of phosphate ions through P amendment. Bolan et al (2003) reported that with addition of KH2PO4 as a source of amendment to the contaminated soils, the plant growth of Brassica juncia increased with application of P amendment in the Cd contaminated soils and also phosphorus application decrease the Cd concentration in plants. The reduction in phyto toxicity of Cd due to phosphate induced immobilization of Cd in soils, 17

which could be explained by precipitation of Cd as Cd(OH) 2 and Cd3(PO4)2.The mechanism involved in phosphate induced Cd adsorption included increase in pH , increase in surface charge, co-adsorption of phosphate and Cd as an ion pair and surface complex formation of Cd on phosphate compound. Ma et al (1994) reacted Pb and synthetic hydroxyl apatite (HA) in the presence of various levels of Al, Cd, Cu, Fe, Ni or Zn at different pH levels and observed a significant reduction in Cd concentration in solution. They suggested that mechanisms for reducing Cd concentration in soil solution were likely the formation of amorphous mixed-metal phosphates (Cd-phosphates or Ca-Pb-Cd phosphates), sorption on apatite surfaces, or ion exchange reactions. Laperche et al (1996) reported that addition of phosphorus may reduce phytoavailbility of Cd through a combination of several mechanisms, such as sorption, precipitation or co precipitation. Valsami-Jones et al (1998) studied the reactions of synthetic HA and natural apatite with aqueous Pb and Cd and found that Cd concentration were reduced in the aqueous solution. They also suggested similar mechanism of formation of amorphous mixedmetal phosphate as well Chen et al (1997) suggested that reduction in aqueous Cd concentration with apatite addition occurs primarily because of sorption mechanism such as surface complexsation and ion exchange rather than precipitation of Cd phosphate. Laperche et al (1997) investigated the effect of different apatite amendments on Pb availability to sudan grass and reported that in the absence of apatite, the Pb concentration in sudan grass were as high as 170 mg Pb kg-1 which got reduced to 3mg kg-1 Basta and McGoven (2004) studied that the evaluation of three chemical immobilization treatments i.e. agriculture limestone (AL), mineral rook phosphate and diammonium phosphate (DAP) to reduce heavy metal (Cd, Zn, Pb) transport in a smelted contaminated soil. They observed that DAP treatment was superior to all other materials for reducing Cd, Zn and Pb concentration. Application of DAP @ 10g kg respectively. Brown et al (2004) reported that with application of phosphorus amendments, the phytotoxicity of Cd was reduced which led to higher plant yield in tall fescue grass and observed that 1 % H3PO4 treatment was the most effective to reduce plant Cd concentration. Hettiarachchi and Pierzynski (2002) studied the effect of various P amendments on plant growth of Sudan grass and Swiss chard. They reported that Cd concentrations in both the crops were reduced significantly by triple super phosphate (TSP) but did not change with the addition of rock phosphate indicating thereby that plant growth was enhanced by the presence of soluble phosphorus source due to formation of mixed metal-phosphates. The lower solubility of these metal phosphates could have restricted metal uptake by plant. Cao et
-1

was most effective

treatment for immobilization of Cd, Pb and Zn with reduction of 94.6, 98.9and 95.8

18

al (2003) also reported that with the application of phosphorus through phosphate rock, the Pb, Zn and Cu concentration in roots of Augustine grass was decreased. Raicevic et al (2005) studied that in situ immobilization of toxic metals, using inexpensive `reactive` amendments, is considered as a simple and cost effective approach for the treatment of soils, contaminated by the presence of heavy metals as compared to ex situ and costly techniques. The stabilization of heavy metals in contaminated soils and ground water by addition of apatite minerals has the potential to be a successful and widely applicable remediation strategy for the case of Cd, Pb as will as for other heavy metals existing in polluted soils. The theoretical analysis of stability, regarding the apatite/Cd or apatite/Pb systems and relevant results of sorption experiments, pointed out two different mechanisms for the immobilization of Cd and Pb by use of apatite to remediate the contaminated soils. Strong antagonism between P and Cd has been reported by Brown et al (2004). Restricted Cd availability due to P application was also confirmed by Panwar et al (1999, 2001) and Datta et al (2007). Matusik at el (2008) study the In situ immobilization of heavy metals in polluted soils using phosphates leads to formation of products which are highly insoluble and thermodynamically stable over a broad pH and Eh range. The highest reduction of cadmium concentration (>99%), owing due to the formation of cadmium phosphates, was observed for all used phosphorus sources within pH range of 6.759.00. Mark et al (2000) suggested that, due to low solubility, the formation of metal phosphates in soils contaminated by metals may represent a cost-effective, sustainable, in situ method for the remediation of metal contaminated soils. A variety of laboratory and field experiments have been carried out to investigate this proposition and these are reviewed. In laboratory studies carried out by the authors, and summarized here, bone meal (ground bone) was used as a phosphorus source. Low concentration bone meal additions to metal contaminated soils resulted in the immobilization of lead, zinc, nickel and copper, and increases in the pH of the soils. The relatively low cost of bone meal means that bone meal amendments could be a cost-effective treatment for metal contaminated soils 2.3.2 Effect of CaCO3 on Cd concentration and its uptake Lime and other alkaline amendments raise pH that leads to a higher affinity between soil and metal species, but also leads to formation of precipitates and secondary minerals that decreases metal solubility and transport (Basta and Mcgoven 2004) Singh and Nayyar (2001) reported that with applied calcium carbonate at the rate of 2.5 percent to the soil, both DTPA-extractable and plant Ni decreased which helped in mitigating the toxic effect of Ni as evidenced by the increase in dry matter yield of cowpea. 19

Increased levels of Ca2+ can decrease the amount of Cd that is assimilated by plants (Larlson et al 2000). Because of their similar size, Ca (II) is almost in distinguishable from Cd (II) (Ochiai 1995). A higher affinity for the essential trace metal Ca results in the decreased uptake of Cd into the plant. A similar relationship exists between P and Cd. John et al (1972) showed that the addition of 1000 ppm of phosphorus to a Cd contaminated soil decreased the concentration of Cd 43% in the roots of oats. Trace metal deficiencies in plants have been associated with increases in heavy metal uptake (Khan and Frankland 1983). Peles et al (1998) concluded that the addition of lime to contaminated soils (essentially increasing the pH) decreased the uptake of heavy metals. In unlimed soils Ambrosia trifida accumulated 13.6 g Cd g-1 of tissue and in limed soils A.trifida accumulated 2.5 g Cd g-1 of tissue. Tlustos et al (2008) reported the influence of the addition of CaO and CaCO3 in contaminated soil containing 7.14 mg Cd kg -1, 2174 mg Pb kg -1, and 270 mg Zn kg-1 on element availability for spring wheat in pot experiment. The ameliorative materials were added into the pots containing 5 kg of soil in amount of 3 g CaO, and 5.36 g CaCO3 per kg of the soil. Soil pH increased to 7.3 in lime treatments compared to 5.7 in control soil. Mobile portion of soil elements (0.01 mol l-1 CaCl2 extractable) dropped by 80% for Zn, 50% for Cd, and 20% for Pb, respectively. In both straw and grains of wheat reduced content of elements was observed in limed pots compared to the control ones. Han and Lee (1996) observed reduction in uptake of Cd and Pb in Raphanus sativa L. var. Paekyong using the application of lime. Soils treated with 1.52 mg kg 1 Cd and 25.37 mg kg 1 Pb, respectively was grown in greenhouse pots and amended with lime at five rates of 0, 0.25, 0.5, 1.0 and 2.0% by dry soil weight. Plants were harvested at 25, 50, and 75 days after sowing and the roots and shoot separated. Liming decreased Cd uptake markedly at its highest level. Xian (1989) reported that exchangeable and carbonate constituted 55 and 11% of total Cd respectively and he also observed that potential bioavailability was strongly controlled by their chemical forms related to solubility and exchangeable and carbonate forms were the most important forms for heavy metal uptake by plant. Ma and Gade (1997) obtained 57 to 100% reduction in water soluble Pb with the application of crushed rock phosphate to a lead contaminated soils. Lee (2004) study the effect of soil amendments, including compost, zinc oxide, calcium carbonate, calcium carbonate mixed with zinc oxide, and calcium carbonate mixed with compost. The amended soils were incubated for six months under 60% of water holding capacity. Following incubation, wheat was grown for four months in greenhouse. Analyses of Cd concentration demonstrated a significant decrease in soil solution concentration and DTPA or EDTA extractable in soils amended with calcium carbonate or calcium carbonate 20

mixed with ZnO (or compost) (p < 0.01). These amendments can significantly reduce the Cd concentration in the grain, leaf and stem, or reduce the total Cd uptake in all parts of wheat species grown in highly contaminated soil amended with calcium carbonate or calcium carbonate mixed with ZnO (or compost) (p < 0.01). 2.3.3 Effect of FYM on cadmium concentration and its uptake The heavy metal concentration in soil and may lead to the phyto toxicity to crop plants. Various organic and inorganic amendments are applied to contaminated soils to reduce the toxic effects of heavy metals in soils as well as plants. Organic compounds like Farmyard manure, biosolids and poultry manure are known to decrease the phytotoxity of different heavy metals like Cd, Cr and Cu etc. FYM is easily available source of organic matter, cost effective. Hiroyuki et al (2010) conducted a field experiment and observed efficiency of cattle waste compost (CWC) in reducing Cd uptake by spinach (Spinacia oleracea L.). Spinach was grown in a field that had been treated by having cattle, swine, or poultry waste compost incorporated into the soil before each crop throughout 4 years of rotational vegetable production. Cadmium concentration was 3438% lower in spinach harvested from the CWCtreated soils than in the chemical fertilizer-treated soil. The risks related to municipal solid waste compost application in comparison to farmyard manure and mineral fertilizers on durum wheat were investigated on a short-term experiment by Lakhdar et al (2008). Compost was applied at 40 t ha -1 and 80 t ha-1 with or without chemical fertilizers. Analogously, farmyard manure was applied at 40 t ha-1. Both compost and farmyard manure improved plant growth and nutrient uptake. However, compost amendment showed more effectiveness, especially at 80 t ha-1. Alternatively, this dose of compost involved an increase of plant copper, cadmium, and zinc concentrations in plant tissues. Metal accumulation did not thwart the enhancement of wheat yield. Putwattana (2010) conduct a pot experiment and study the effect of cow manure on the cadmium uptake and dry matter yield and result showed an increase in dry biomass production by factors of 4.7 and 1.7 in plants grown in soil supplemented with cow manure (20% w/w) and silicate fertilizer (20% w/w), respectively by Ocimum basilicum (sweet basil) grown on in Cd contaminated soil (20 mg/kg Cd) Singh et al (1992) Studied that in a screen house experiment that Cd concentration in plants decreased with the addition of organic matter at all cadmium levels. Dahiya et al (1987) studied the effect of Cd and FYM on dry matter yield of maize and found that FYM application increased significantly dry matter yield of maize by decreasing the concentration of Cd in maize plants. Khurana and Kansal (2000) studied the bio-availability of Cd to maize crop as influenced by Cadmium and Farm Yard Manure. They 21

found that addition of FYM increased organically and oxide bound but decreased the exchangeable + water soluble and carbonate bound fraction. Increase in comparatively insoluble fractions of Cd helped to mitigate the toxic effect of Cd as evidenced by increase in dry matter yield. 2.4 Transformation of heavy metals in soils Increasing amount of pollutant elements are entering the soil plant system through anthropogenic activities, sewage waste and fertilizers. Once these elements specially Cd .,Pb and Ni (found commonly in soils in toxic amounts around the industrial towns) enter the soil plant system, they interact with its organic and inorganic constituents. Then they get transformed into various forms like soluble, exchangeable bound to carbonates, Fe and Mn oxides, organic matter and residual (Bell et al 1991). Sequential procedures have been used to identify these forms (McLaren and Crawford 1973; Cottenie et al 1979). Researchers have often noted that metal added to soils as reagent grade inorganic salts were more readily available to plants than those present in sewage sludge. Once these heavy metals are incorporated into soil, their extractability appears to change with time indicating a possible change of their chemical form. So metal toxicity depends on chemical associations in soils. For this reason determining the chemical form of a metal in soils is important to evaluate its mobility and bioavailability. Ghafoor et al (2008) study the effect of different inorganic amendments on fractionation and availability of Cd to. Inorganic amendments viz. lime, gypsum, diammonium phosphate (DAP) and potassium dihydrogen phosphate (KH2PO4) was used at different rates. Maximum reduction in Cd in grain (75%) and straw (64%) was observed where KH2PO4 were applied at 2000 mg kg-1 followed by the treatment, where DAP was applied at the rate of 2000 mg P kg-1. Lime application significantly decreased Cd concentration by 54% and 64% in straw and grain as compared to control, while decrease with gypsum 41% in straw and 61% in grains Kuo et al (1983) observed that 30-60 per cent of total Cd was found to be in exchangeable fraction (Soluble in MgCl2). This per cent was found to be much greater than that of other elements including Fe and Mn. Most of the Cu, Zn and Mn were present in an oxalate extractable fraction but only small amounts were extractable by citrate dithionitebicarbonate (CDB) solution. Cu, Zn and Mn were strongly associated with amorphous Fe oxides with only small amounts being occluded in crystalline iron oxides. It appeared that Cd, Ni and Zn were shifting to residual forms. The occurrence of metals in residual forms in the soil after sludge incorporation, contributed to the lack of metal movement in the soil profile. Berthet et al (1984) assessed that levels of exchangeable and easily reducible Cd were very low in sewage sludge than in the soil. Levels of Cd associated with organic matter were 22

high and moderately reducible forms were frequent in both soils and sludge. Almost half the Cd in sewage was in stable forms unavailable to plants.Cd concentration in the plants being generally higher in treatments than controls. Lake et al (1984) reported that following the application of municipal sewage sludge to agricultural soil, the metals were being predominantly associated with solid phases, soluble exchangeable species generally represent < 10 per cent of total metals. Speciation in sludge amended soil initially reflected that of sludge itself although changes with time had been observed. Gibson and Farmer (1986) studied various fractions of cadmium and zinc and found that 32 per cent of Cd was associated with the exchangeable + carbonate fractions, 29 per cent of zinc with the organic fraction, 42 and 46 per cent of the total Cd and Zn were found in the residual form. A sequential extraction procedure incorporating I M KNO3, 0.5 M KF, 0.1 M Na2P207, 0.1 M EDTA and I M HNO3 had been used to fractionate metals in to exchangeable, adsorbed, organically bound, carbonate bound, and sulphide forms respectively (Stover et al 1976). Carbonate constituted 49 per cent of Cd, 32 per cent of Ni and 61 per cent of Pb on average while 35 per cent of Cu was present in sulphide form, 50 per cent of Zn was organically bound. For Cd, Cu, Pb and Zn adsorbed and exchangeable fraction accounted for only 17 per cent. In sandy loam soils treated with sewage sludge, Sposito et al. (1982) found that Zn, Cd and Pb were mainly present in carbonate form, Cu in organic form and Ni in sulphide form. Irrespective of sludge application rate, exchangeable metals averaged between 1.1 and 3.7 per cent. Soon (1981) revealed that soils treated with sludge increased the amount of exchangeable cadmium but reduced the amount of complexed Cd compared with fertilizer soil. Cadmium retention by cation exchange became more dominant with the increasing amount of Cd in the soil. He postulated that with addition of 50 g Cd g-1, its precipitation as CdCO3 was at pH>7.6. Cadmium adsorption increased with increasing pH. The difference in Cd adsorption between different soil treatment were attributed mainly to the change in soil pH (6.9 to 7.9) induced by sludge application. Alekseev and Zyrin (1982) found that water soluble and exchangeable forms of cadmium were transformed into more stable form with time. They also observed that mobility of cadmium and its availability for plants were substantially greater in acid soil than in non calcareous, neutral and alkaline soils. According to Silviera and Sommers (1977), water soluble and exchangeable metals (Cu, Zn, Cd and Pb) comprised a small per cent of the total metal concentration in the sludge and in soil-sludge mixture incubated for 7 to 28 days. The DTPA extractable fraction of the total Cu, Zn, Cd increased with time but DTPA extractable Pb remained unaffected. Emmerich et al (1982) employed sequential extraction procedure for applied sewage sludge 23

and uncontaminated soil samples. They reported that less than 35 per cent of each metal (Cu, Ni, Zn Cd) in sewage sludge was in the residual form but the soils contained greater than 65 per cent of each metal studied in this form. It appeared that Cd, Ni and Zn were all shifting to residual forms. The occurrence of metals in the stable organically bound, carbonate and residual forms in the sludge coupled with a shift towards the more stable form (residual) after soil incorporation, contributed to the lack of metal movement in the soil profile. Soon and Bates (1982) studied the chemical pools of cadmium, nickel and zinc in polluted soils and extracted the soil with 1M ammonium acetate to remove soluble plus exchangeable metals, with 0.125 M copper acetate to remove complexed metals and with I M HNO3 to dissolve chemisorbed or occluded metals and precipitates such as oxides and carbonates. Expressed as a per cent of metal so extracted, exchangeable Cd>Zn and Ni, complexed Cd and Zn >Ni, and Ni>Zn> Cd in the acid soluble pool. With a few exception (soils with high organic matter content or low pH) at least 50 per cent of extracted metal was in acid soluble form. But when soil 1 and 2 were treated with CdCl 2 (samples I-A and 2-A) more of added Cd was soluble and exchangeable compared to treatments which received similar amount of Cd from sewage sludge. Xian and Shokohifard (1989a) investigated the effect of pH on chemical forms and plant availability of heavy metals in three polluted soils. Heavy metals were portioned in to five operationally definite chemical fractions: exchangeable, carbonate, FeMn oxides, organic and residual. When soils pH values were decreased from 7.0 to 4.55, levels of Cd, Zn and Pb in exchangeable forms increased but decreased in carbonate and Fe-Mn oxides forms. Their levels in organic and residual forms were unchanged. Khan and Frankland (1983) reported that a very high proportion of Cd and Pb added to the soil became water insoluble fraction within one hour of contact with the soil, although most of the water insoluble fraction was EDTA soluble. Miller and McFee (1983) sequentially extracted the heavy metals with I M KNO 3, Sodium pyrophosphate, 0.1 M EDTA, 0.1 M hydroxyl amine hydrochloride and 0.01 N nitric acid, 0.27 N sodium citrate + .1N sodium bicarbonate + 0.25 g Na2S2O4, I N HNO3, and HNO3 + H2O2 for removing water soluble exchangeable, organically bound, carbonate / non crystalline Fe occluded Cd, Mn oxides occluded, crystalline Fe oxides occluded, sulphide and residual to study the distribution of cadmium, zinc, copper and lead in soils of industrial north western Indiana. The result showed large amounts of relatively labile metals associated with exchange site (KNO3 extractable), 23, 10, 1 and 8 per cent of total Cd, Zn, Cu and Pb respectively, bound by soil organic matter (Na2P2O7 extractable : 21, 33, 24 and 41 per cent of total Cd, Zn, Cu and Pb) and associated with carbonates and/or non crystalline Fe oxides (EDTA-extractable : 12, 8, 26 and 28 per cent of total Cd, Zn, Cu and Pb). Minimal amount of the metals were within small amount of crystalline Fe and Mn oxides present in these soils. 24

Non extractable (residual) metals amounted to 26, 32, 23 and 41 per cent of total Cd, Zn, Cu and Pb. Xian (1989b) reported that the exchangeable fraction contained 55 and 13 per cent and carbonate fraction 11 and 10 per cent of the total Cd and Zn respectively. The highest amount of zinc (42 per cent) was detected in residual form. Cadmium distribution to organic and residual was small especially in paddy soils. Metals level in cabbage plants were in accordance with their levels in the soil. He also observed that potential bioavailability of heavy metal was strongly controlled by their chemical forms related to solubility and exchangeable and carbonate forms were the most important forms for heavy metal uptake by plants. Okamoto et al (1990) found that the heavy metals were mostly in the carbonate and Fe-Mn oxides forms with low mobility and availability in soil amended with sewage sludge .When the sludge application was interrupted, and the soil was treated with acid fertilizers, the chemical forms of the heavy metals changed from carbonate or Fe-Mn oxides to soluble and exchangeable forms, with an increase in availability and mobility due to decrease in soil pH Dudka and Chlopecka (1991) observed that mobility and plant availability of zinc and cadmium in sludge treated soil were associated mainly with the exchangeable fraction. Jeng and Singh (1993) studied partitioning and distribution of cadmium and zinc in selected cultivated soils in Norway. These soils were analysed by sequential extraction to isolate five fractions of Cd and Zn. On average, 47, 4, 33 and 5 per cent of the total Cd fractions were classed as weakly adsorbed (F1), adsorbed (F2), strongly adsorbed (F3) and very strongly adsorbed (F4) forms of elements respectively. 11 per cent was regarded as incorporated in resistant mineral (F5). The relatively high proportion of F1 fraction indicated that much of Cd under these conditions was available to plants. For Cd, F1 and F3 were directly correlated with organic carbon and total iron. Organic carbon also seemed to be important in retaining F4, where as F5 correlated best with clay. Pierzynksi and Schwab (1993) studied the sequential fractionation of Cd. Soil samples were extracted with 0.5 M KNO3, deionized water three times, 0.5 M NaOH, 0.05 M Na2EDTA and 4 M HNO3. The sum of KNO3 and water extractable metals were assumed to represent the most liable metal pool, the NaOH fraction were assumed to represent the organically bound pool, EDTA to represent metal from inorganic precipitates and HNO3 fraction was residual. They further studied the effect of lime stone, cattle manure, poultry manure, N-vivo, K2HPO4 and (NH4)2HPO4 on these fractions. Lime stone treatment was clearly the most effective in reducing bioavailable (KNO3, H2O and NaOH) fraction of Zn and Cd. But NaOH-Cd was significantly increased by cattle manure, poultry manure and K2HPo4 treatment as compared to control. Lime stone treatment significantly increased EDTA-Cd fractions as compared to control. Corresponding to reduction in NaOHCd with increasing 25

lime stone rates, was a significant increase in EDTA-Cd suggesting an increase in precipitated Cd in the presence of lime stone. Significant increase in NaOH-Cd observed with addition of manure might be attributed either to Cd additions with manure or partitioning of Cd into organic fractions with increasing organic additions. Ramos et al (1994) employed sequential extraction procedure to partition the Cd, Pb, Cu and Zn in soils representing three different areas in Spain, i.e. marshes, stabilized sand and mine. Marshes had the greatest total levels of four metals due to the influence of a mine located 40 km away while the stabilized sand was non polluted. The bioavailable fraction of Cd represented > 50 per cent of total Cd in the soils. The total amount of Cu, Pb, Cd and Zn in the soils and their distribution in the five fractions depended on the total metal content, soil type and soil properties. Mobilities of the metals were in the order of Cd > Zn > Pb > Cu. The distribution of the chemical forms of Cd and Cu in four inceptisols was studied using a sequential extraction procedure by Ahumada and Schalscha (1994). The metals were separated into five fractions: water soluble, exchangeable, reducible, DTPA extractable and insoluble organically bound forms. Cadmium was recovered mostly in the insoluble organic matter fraction. The addition of extra Cd did not affect this distribution. Copper was recovered mostly in the reducible and the Fe and Mn oxides bound forms. By increasing the redox potential of soils from 150 to +50 mV and then to 300 mV, the proportion of exchangeable metals decreased while the proportion of the reducible form increased. The proportion of plant available Cd was unaffected and the distribution of Cu forms was unchanged. Asami et al (1995) fractionated soil Cd, Zn, Pb and Cu in 38 soil samples from 11 soil profiles of industrially polluted and nearby non polluted areas in Japan. On average 45 per cent of Cd was present in CaCl2 (Ca) soluble fraction where as corresponding values for other metals were less than 10 per cent. The per cent of each metal in the CA fraction followed the order Cd>Zn>Pb>Cu. The same order was observed for acetic acid soluble fraction. Approximately 30 per cent of total Pb and Cu and only 10 per cent of total Cd and Zn were present in the pyrophosphate soluble fraction. Approximately 20 per cent of total Zn or Pb and 10 per cent of Cd or Cu were present in the free oxides fraction. Only 20 per cent of Cd and between 40-50 per cent of other three metals were present in the residual fraction. The result showed that Cd was more labile than other metals. Taylor et al (1995) conducted fractionation of residual cadmium, copper, nickel, lead and zinc in soils that had received previously two types of sludge at two different rates, fertilizer and control. The content of cadmium, copper, nickel, lead and zinc fractions in previously sludge amended soils were governed by total content of these metals in the sludges applied and by the rate of sludge application. The contents of these metals were higher in soils that received the Chicago sludge as compared to that received the Hunstsville sludge. 26

Furthermore, soils that received 20 t/ha/year of sludge for 5 years generally had higher levels of these metals than those receiving a single dose at 100 t ha-1. The per cent of total content in water soluble and exchangeable forms was very low. The application of sludge tended to reduce the residual fraction and increased the organic and carbonate fraction. Overall, the predominant forms of metals in the sludges were as Cd, Ni, Pb and Zn carbonate and Cu organic fractions. Sadamoto et al (1995) assessed the heavy metal pollution in terms of chemical forms especially Cu, Zn and Cd. Extraction with H2O2 and acetic acid (2.5 per cent v/v) recovered greater quantities of metal than 0.1M potassium pyrophosphate. Oxalic acid + UV radiation incompletely extracted metals occluded by Fe oxides whereas oxalic + ascorbic acids reduced the oxides enabling extraction of the occluded metals. Acetic acid (2.5 per cent v/v) extracted metals sorbed by geothite and gibbsite. The residual fraction comprised 50 per cent of soil Cu, 60-70 per cent of soil Zn. Cadmium however was found predominantly in exchangeable forms. Liu et al (1997) studied the desorption mechanisms and chemical forms of Cu and Cd in major Taiwan agricultural soils, high in organic matter and Fe-Mn oxides contents to evaluate the fate of heavy metals. The result showed that Cu was mainly associated with Fe-Mn oxides (29.4 per cent), carbonate (23.8 per cent) and organic matter (17.0 per cent) while Cd existed mainly as water soluble and exchangeable forms (68.6 per cent). All the results showed that Cd pollution might had greater impact on plants and environment than Cu pollution. Singh et al (1998) characterised the surface soils near some disposal sites in Belgium for total metal contents and various fractions. Residual fractions were low compared to total content (2-4 per cent for Cd, 25-35 per cent for CO, 7-18 per cent for Mn, 4-22 per cent for Zn, 11-42 per cent for Pb). High metal concentration in the acid extractable and reducible fractions indicate pollution hazards. Singh et al (1995) extracted the heavy metals with DTPA and NH4NO3 at different pH values in a clay loam and loam soil. They found that the DTPA extractable and NH4NO3 extractable Cd had decreased with increasing soil pH and the effect was more pronounced with NH4NO3 extractable Cd. Both extractants were found equally effective in relation to Cd concentration in plants. The review of literature presented in the preceding pages indicated that application of metal contaminated sewage water, sewage sludge, solid wastes and fertilizers and caused significant contamination of agriculture soils with heavy metals including cadmium. Review also suggested that separation of various forms in soils had been useful in studying the retention and release of elements by the soil to the plant and was particularly more important for pollutants added through waste materials. The addition of amendments influenced the chemical forms of cadmium in a soil and therefore, helped to reduce the toxicity of cadmium. Studies pertaining to the use of farm yard manure, CaCO3 and phosphorus were rather scanty.

27

Therefore present study was undertaken to investigate the effect of these amendments on transformations of this element to evaluate its bioavailability.

CHAPTER III MATERIAL AND METHODS Keeping in view, the objectives mentioned in chapter-1, the materials used and the methods followed during the course of study have been reported under the following heads. I) Surveys Studies II) Screen house studies III) Laboratory studies I) Survey studies Ludhiana city was founded on a ridge of Buddah Nallah, which once was a bed of the river Satluj. The urban area is lying between 30o 51'10" to 30o 57' 20"N latitude and 75o 46'00" to 75o56'20"E longitude, the average height above mean sea level is 247 m. Previously the urban area was confined south of it but due to burgeoning population, the low lying area between Buddah Nallah and the river Satluj is fast getting urbanized In order to determine pollution potential, surface (0-15cm) soil samples receiving waste water irrigation at a distance of 50, 250, 500, 750 and 1000 meters (5 sites) were collected laterally on either side (Right and left) along the Buddah Nallah from each village (six in number) which were approximately 4-5 kilometers from each other with the help of global positioning system in the month of December. The six villages selected for this purpose were Bhamian khurd, Saidan, Saleem tabri, Pratapsinghwala, Talwara and Jain pura. It was not possible to collect samples from all the sites of the all the villages due to built area. Details of sampling sites from each village is mentioned in table 4.1 Thus, in total, 53 soil samples were collected for this purpose. Surface (0-15cm) soil samples were also collected from nine villages namely Bhamian Kalan, Baranhara, Balloke, Ladian Khurd, Bagha Khurd, Jhamat, Ayali khurd, Malikpur, Rajowal where no sewage water was applied. These sampling sites are schematically shown in map (fig 1) II) Screen house studies Bulk soil sample (0-15 cm surface) was collected from sewage irrigated soil near to the Buddah Nallah (Ludhiana). Soil was air dried ground and passed through a 2 mm stainless

28

steel sieve. The physico chemical characteristics of processed soil are given in table 3.1. The processed soil was thoroughly mixed and used for screen house studies.

Table 3.1. Physico-chemical characteristics of the soil. Characteristics pH(1:2)* EC(ds m-1)* Organic Carbon (%) Calcium carbonate (%) Macro nutrients (Kg ha-1) N P K DTPA extractable (mg kg-1 soil) Cadmium Zn Fe Mn Cu Mechanical Composition Sand (%) Silt (%) Clay (%) Texture *( 1:2, soil: water suspension) Experiment: 1 Treatments Level of cadmium (6) Levels of amendments (7) : 0, 2.5, 5, 10, 20, 40 mg kg-1 soil : 0, 2.5%, 5 % (Calcium carbonate), 1%, 2% (FYM) and 20, 40 mg P2O5 kg-1soil (CaH2PO4) Replications 29 : 4 75.2 15.0 9.80 Loamy Sand 0.36 1.38 12.6 5.40 1.00 278.4 15.7 137.8 Contents 7.02 0.54 0.24 0.20

Crop Experimental Design No .of pots

: Chalai(Pig weed-Amaranthus tricolor) : Completely randomized factorial design : 6 x4x 7 =168 pots

Treatment imposition and fertilizer application: Bulk surface (0-15cm) soil sample was collected from the sewage irrigated soil. This sample was air dried ground and sieved. Nine kg of this processed soil was taken in each polythene pot. Cadmium at the above rates were applied through cadmium chloride and amendments (Calcium carbonate: 2.5 and 5 %, FYM: 1 and 2% Phosphorus: 20 and 40 mg P2O5 kg-1 soil through CaH2PO4. The soil in the pots was subjected to alternate wetting and drying cycles for 30 days to attain equilibrium. Thereafter, representative soil samples from each pot was taken, processed and stored in polythene bags for chemical analysis. After application of basal recommended doses of N, P and K, the seeds of Pig weed (Amaranthus tricolor) was sown in pots under optimum moisture conditions. After germination, thinning was done to maintain 6 plants. The soil in the pots was irrigated as and when required. The crop was harvested after 45 days of sowing. Shoot, root and soil samples were collected from each pot. The shoot and root samples were analyzed for total Cd as well as for Cu, Fe, Mn and Zn while soil sample was be analyzed for DTPA-Cd .The upper threshold value of Cd in soil and crop was determined by employing suitable method. Harvesting, collection and processing of shoot and root samples: The shoot samples were harvested with the help of stainless steel blade. For harvesting and collection of root samples the polythene bags were taken out from the pots. The soil was washed with tap water from each polythene bag. The roots were separated and put in the stainless steel sieve and washed with tap water pressure, water jet, acidified deionized water and finally with double distilled water. The harvested shoot samples were also washed with acidified deionized water, distilled water and with double distilled water. The washed shoot and root samples were first air dried by keeping them in paper bags and then in oven at 652 oC. Thereafter, the shoots and roots were weighed for dry matter yield. Shoots and roots of the plants were ground by stainless steel grinder and stored in polythene bags separately. Cadmium as well as Fe, Mn, Zn and Cu were analysed in shoots and roots. Post harvest soil sampling: After harvesting the crop, composite soil samples from each pot were taken with stainless steel tube augar. These soil samples were air dried, ground, sieved and stored in polythene bags for their chemical analysis. 2 Laboratory studies Sequential extraction of soil samples: A seven step sequential extraction procedure given in Table 3.2 was applied to evaluate the association of metals with soil constituents. The different chemical forms of fraction of Cd were exchangeable + water soluble, carbonates, organic mater 30

complexed, Mn-oxide bound, occluded in amorphous and crystalline Fe oxide and residual mineral fraction. Several methods of fractionation of heavy metals have been proposed by several workers (Chao 1972; Gupta and Chen 1975; Tessier et al. 1979; Chao and Zhou 1983; Shuman 1985). Here sequential extraction procedure adopted by Singh et al (1988) was used with some modifications to simulate the conditions, which is described in Table 3.2. Reagents used in fractionation scheme were selected from those cited in the literature as being selective for specific chemical from in soil. All procedures were carried out in triplicate. Five gram of each soil sample was taken in centrifuge tube then centrifuged and filtered after fulfilling the essential conditions such as shaking, boiling, digestion etc. the soil residue was washed twice with double distilled water. Residual metal analysis: Residual from C-FeOX extraction was washed twice with double distilled water. It was subsequently dried and ground to pass through a 2mm stainless steel sieve. A 0.5g sample was digested with 10 ml concentrated HF and 2-3 ml concentrated HClO4 remaining material was taken up in 6N HCl and volume was made to 25 ml. Analysis methods: The following methods were used for soil analysis: 1) 2) 3) 4) 5) Mechanical analysis was done as per International pipette method (Piper 1950). pH and electrical conductivity were determined in 1:2 soil water suspension with the help of glass electrode pH meter and conductivity meter bridge respectively. Organic carbon was estimated by Walkley and Blacks rapid titration method as described by Jackson (1973). Calcium carbonate was estimated by following the method of Puri (1949). DTPA extractable polyvalent transition metals were extracted by using the procedure of Lindsay and Norwell (1978) and estimated on AAS. Total heavy metals: A sample of soil was ground to pass through 2 mm stainless steel sieve. A 0.2 gram sample was digested as per method described in residual fraction (Tessier et al 1979).The amount of other metals and Cd in each fraction were estimated by atomic absorption sepectrophotometry (AAS). Analysis of (FYM): One gram of oven dried FYM sample was digested in 20 ml of distilled concentrated nitric acid at low heat on a hot plate to oxidise organic matter and digested the mixture with 20 ml of perchloric acid until dense white fumes of acid appeared. Diluted the digest with double distilled water to known volume Analysis of roots and shoots: In order to determine Cd and other heavy metals in shoots and roots, 0.5g of ground and well mixed plant material was digested in a diacid mixture of nitric acid and perchloric acid (4:1). After digestion, the volume was made to 25 ml with double distilled water, filtered and stored in well washed plastic bottles. All the estimations in the aliquot were made by using Atomic Absorption Spectrometer. 31

Statistical analysis: Factorial CRD was employed to study the effect of different treatments in various experiments (Panse and Sukhatme, 1967). The analysis was carried out with the help of a computer. The effects of treatments were compared with the help of interaction with CD (Critical Difference). Table 3.2 a. Sequential extraction methods used for post harvest distribution of Cd Sr. No. 1 Fractions Solution Soil (g) 5 Solution (ml) 20 Conditions Shake 25C 2 hrs Reference at Shuman (1985)

Exchangeable + 1 M Mg(NO3)2 Water soluble (EXCH+WS) Carbonates (CARB)

1 M NaOAC 5 (pH 5.0 CH3COOH) NaOCl 5

20

Shake 25C

hrs

at Tessier et al (1979)

Organically (OM)

bound 0.7 M (pH 8.5)

20

Shake 30 min in Shuman boiling water bath, (1983) stir occasionally, repeat extraction

Mn-Oxides (MnOX)

bound 0.1 M 5 NH2OH.HCl Sol. in 0.01 M HNO3 (pH 2.0)

25

Shake 30 min

Chao (1972)

Amorphous Iron 0.25 Oxide bound (A- NH2OH.HCl FeOX) 0.25 M HCl

M 5 +

25

Shake 30 min in Chao and boiling water bath, Zhou (1983) stir occasionally 30 min in boiling Shuman water bath, stir (1982) occasionally

Crystalline Fe-oxide 0.2 M (NH4)2 5 bound (C-FeOX) C2O4 + 0.2 M H2C2O4 (pH 3.0) + 0.4 M ascorbic acid

25

32

Table 3.3: Chemical composition of farm yard manure (on oven dry weight basis) Characteristics Organic carbon (percent) pH* EC (dS/m)* Total metals content (mg kg-1 soil) Iron Manganese Copper Zinc Cadmium *1:4 farm yard manure: water suspension Content

20.18 8.0 3.16 2140 280 9.40 72.4 0.38

33

CHAPTER IV RESULT AND DISCUSSION The experiment results pertaining to the present investigation have been presented and discussed under the following heads.

4.1 4.1.1
4.2

Survey studies: Delineation, demarcation and mapping of cadmium polluted soils sewage channel (Buddah Nullah) in Ludhiana district.
Influence of different levels of Cd and and various amendments on 4.2.1 Dry matter yield of shoots of chalai (pig-weed) 4.2.2 Dry matter yield of roots of chalai (pig-weed) 4.2.3 Cadmium concentration in shoots and shoots of chalai (pig-weed) 4.2.4 Cadmium uptake in shoots and shoots of chalai (pig-weed) 4.2.5 DTPA extractable Cd in equilibrated soils 4.2.6 DTPA extractable Cd after the harvest of crop

along a

4.3 4.4

Micronutrient cations concentration in shoots and roots of chalai (pig-weed) Upper critical level of Cd in Soil Plant

4.5

Effect of various amendments on the transformations of Cd in soils (laboratory studies)

4.1 Survey studies

Study Area

Buddah Nallah, a narrow unlined canal, is the citys sole surface water resource. It originates from Chamkaur Sahib (district Ropar) and merges in the River Sutlej. It is an important drainage line of Ludhiana district, receives waste water from diverse type of industries such as machine tools, electroplating, bicycles and woolen & hosiery manufacturing units. The daily disposal of such waters, which are both of industrial and domestic origin, is quite high. The large volume of domestic and industrial waste water has converted the canal to a virtually sewage drain. In the late nineteenth century, it was a clean water stream as reported in the gazetteer of 1904.
4.1.1 DTPA extractable cadmium: Diethylene triamine pentaacetic acid (DTPA) and more precisely 0.005M DTPA + 0.01MCaCl2+ 0.1M tri ethanol amine (pH 7.3) has been used to measure available form of Cd. Elevated concentration of DTPA extractable Cd was found in sewage fed soils as compared to normal soils at all the locations. Overall mean (average of 10 34

values) DTPA extractable Cd in surface sewage irrigated soils of Bhamian khurd, Saidan, Saleem tabri, Pratapsinghwala, Talwara and Jain pura were 0.11, 0.21, 0.18, 0.18 and 0.13 mg kg-1 soil respectively irrespective of distance from the polluting point indicating high accumulation in surface soils (Table 4.1). The respective mean values of DTPA extractable Cd in the surface layer in tubewell irrigated soils of of Bhamian Kalan, Baranhara, Balloke, Ladian Khurd,Bagha Khurd, Jhammat , Rajowal, Malikpur, Ayali Khurd were 0.06, 0.04, 0.06, 0.02, 0.02, 0.04, 0.02, 0.02 and 0.02 mg kg-1 soil, respectively. When mean value of DTPA-Cd for all the samples (53 in number) of surface sewage irrigated soils was taken, it was found to be 5.2 times greater than the normal soils. The increase in Cd content of soils with continuous application of sludge has been reported by many workers (Singh et al 1985; Mapanda et al 2005; Rattan et al 2005; Liu et al 2005; Mitra and Gupta 1999; Zhang et al 2008; Siebe 1998; Aghabarati et al 2008). 4.1.2 Lateral distribution of Cd on either side of Budah Nallah: The data in Tables (4.1) revealed that mean DTPA extractable Cd in surface sewage irrigated soils was highest near the proximity and decreased as the distance increased laterally (Table 4.1). In general it was highest at 50 meter distance on either side of Buddah Nallah and decreased as the lateral distance on either side of the sewage channel increased to 250, 500, 750 and 1000 meters respectively. Perusal of data in table revealed that at village Bhamian khurd, the amount of DTPA extractable Cd in soil sample receiving sewage irrigation collected at 50 meter away on right side of the sewage channel was 0.14 mg kg-1 soil which decreased to 0.12, 0.10 and 0.08 mg kg-1soil in soil samples collected at a distance of 250, 500 and 750 meter. Similar trend was observed for other sites /location of various villages. Such behavior of enrichment is due to farmers practice of growing vegetables near the proximity of sewage channel which are largely and preferentially irrigated with sewage water laden with heavy metals including cadmium. Secondly, it has been noticed that sampling area at the time of sampling up to 100 meters on either side get submerged with water off and on due to overflow of water during rains .

35

Table 4.1: Cadmium DTPA and total cadmium concentration around Buddah Nallah along with their geographical locations Distance from S no 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 Ludhiana Lattitude N-3055.217' N-3055.305' N-3055.377' N-3055.480' N-3055.207' N-3055.160' N-3055.102' N-3055.047' N-3055.118' N-3055.078' N-3055.012' N-3054.995' N-3055.131' N-3055.181' N-3055.256' N-3055.340' N-3055.390' N-3055.718' N-3055.751' N-3055.808' N-3055.860' N-3055.980' N-3055.718' N-3055.524' N-3055.613' N-3055.762' N-3055.882' N-3055.934' N-3055.507' N-3055.454' N-3055.427' N-3055.345' N-3055.220' Soil Samples Village Name Bhamian khurad Bhamian khurad Bhamian khurad Bhamian khurad Bhamian khurad Bhamian khurad Bhamian khurad Bhamian khurad Sadian Sadian Sadian Sadian Sadian Sadian Sadian Sadian Sadian Saleem tabri Saleem tabri Saleem tabri Saleem tabri Saleem tabri Saleem tabri Partap sinhg wala Partap sinhg wala Partap sinhg wala Partap sinhg wala Partap sinhg wala Partap sinhg wala Partap sinhg wala Partap sinhg wala Partap sinhg wala Partap sinhg wala

Buddah Nallah
50 m 250 m 500 m 750 m Mean 50 m 250 m 500 m 750 m Mean 50 m 250 m 500 m 750 m Mean 50 m 250 m 500 m 750 m 1000 m Mean 50 m 250 m 500 m 750 m 1000 m Mean 50 m 50 m 250 m 500 m 750 m 1000 m Mean 50 m 250 m 500 m 750 m 1000 m Mean

DTPA -Cd 0.12 0.12 0.10 0.08 0.11 0.14 0.10 0.10 0.08 0.11 0.16 0.12 0.1 0.1 0.12 0.18 0.14 0.12 0.12 0.1 0.12 0.24 0.2 0.22 0.18 0.14 0.19 0.28 0.22 0.22 0.18 0.14 0.14 0.17 0.26 0.22 0.16 0.16 0.14 0.17

Total Cd 1.64 1.60 1.50 1.54 1.57 1.72 1.6 1.54 1.28 1.54 1.84 1.64 1.6 1.62 1.68 1.76 1.54 1.52 1.46 1.24 1.44 3.24 3.12 3.1 1.9 1.84 2.49 3.44 3.02 2.76 2.42 2.22 2.2 2.40 3.22 2.88 2.34 1.98 2.32 2.38

Longitude E-07554.781' E-07554. 809' E-07554. 841' E-075 54. 868' E-07554. 779' E-07554. 815' E-07554. 852' E-07554. 846' E-07553.662' E-07553.670' E-07553.687' E-07553.714' E-07553.664' E-07553.676' E-07553.690' E-07553.705' E-07553.711' E-07550.653' E-07550.632' E-07550.581' E-07550.571' E-07550.558' E-07550.823' E-07547.527' E-07547.498' E-07547.523' E-07547.526' E-07547.530' E-07547.529' E-07547.534' E-07547.525' E-07547.510' E-07547.497'

36

34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62

N-3056.275' N-3056.338' N-3056.409' N-3056.507' N-3056.590' N-3056.260' N-3056.240' N-3056.170' N-3056'120' N-3056.098' N-3056.551' N-3056.617' N-3056.676' N-3056.735' N-3056.822' N-3056.525' N-3056.433' N-3056.352' N-3056.272' N-3056.198' N-3054'57.04" N-3056'14.21." N-3056'26.19" N-3056'49.19" N-3057'01.34" N-3054'57.04" N-3057'19.43" N-3055'37.50" N-3055'02.49"

E-07546.182' E-07546.240' E-07546.267' E-07546.310' E-07546.360' E-075 46. 160' E-075 46. 139' E-075 46.057' E-075 46. 048' E-074 46. 029' E-07544.920' E-07544.925' E-07544.921' E-07544.920' E-07544.924' E-07544.911' E-07544.872' E-075 44.835' E-07544.805' E-07544.770' E-07556'47.67" E-07546'58.18" E-07548'25.40" E-07547'17.13" E-07546'03.80" E-07545'44.17" E-07545'30.54" E-07544'38.53" E-07545'57.51"

Talwara Talwara Talwara Talwara Talwara Talwara Talwara Talwara Talwara Talwara Jain pura Jain pura Jain pura Jain pura Jain pura Jain pura Jain pura Jain pura Jain pura Jain pura Bhamian kalan Baranhara Balloke Ladian khurd Bagga khurd Jhammat Rajowal Malikpur Ayali khurd

50 m 250 m 500 m 750 m 1000 m Mean 50 m 250 m 500 m 750 m 1000 m Mean 50 m 250 m 500 m 750 m 1000 m Mean 50 m 250 m 500 m 750 m 1000 m Mean

Mean

0.22 0.2 0.16 0.16 0.14 0.17 0.2 0.2 0.18 0.2 0.14 0.18 0.18 0.14 0.14 0.1 0.06 0.11 0.2 0.16 0.14 0.08 0.1 0.12 0.06 0.04 0.06 0.02 0.02 0.04 0.02 0.02 0.02 0.03

2.24 1.94 1.88 1.9 1.72 1.86 2.36 2.2 1.84 1.76 1.8 1.90 1.98 1.88 1.72 1.46 1.38 1.61 1.86 1.8 1.64 1.26 1.34 1.51 0.98 0.84 1.28 1.02 1.1 1.25 0.81 0.58 0.78 0.86

4.1.3 Total Cd: In general, high concentration of total cadmium was observed in the polluted soils at all sites. Data in tables (4.1) showed that the content of total Cd decreased as the lateral distance increased being maximum at 50 meter and minimum at 1000 meters at all the sites on either sides of sewage drain (Table 4.1). On right side of Buddah Nallah it varied from 1.64 to 1.54, 1.76 to 1.24, 3.24 to 1.84, 3.02 to 2.2, 2.24 to 1.72 and 1.98 to 1.38 mg kg-1 as the distance increased from 50 to 1000 meters with a mean value of 1.57, 1.68, 2.49, 2.40, 1.86 and 1.61 mg kg-1 soil in sewage irrigated soils of Bhamian khurd, Saidan, Saleem tabri, Pratapsinghwala, Talwara and Jain pura respectively. Same trend was observed on left side of Buddah Nallah. Maximum concentration of total cadmum was observed in village Saleem tabri followed by Pratapsinghwala, Talwara, Saidan, Jain pura and least in Bhamian khurd.

37

Fig: 1

38

The increase in total Cd content with sewage water irrigation was dependent upon the rate of loading and chemical composition of sewage water as well as in situ properties of soils (Williams et al 1980; Thakur and Kansal 1992). Elevated concentration of total Cd in the soils of industrial towns of Punjab might also be attributed to the atmospheric fall out of heavy metals since these lie in close proximity to industrial activities. Griffiths and Wardsworks (1980). In order to demarcate Cd polluted soils, guidelines based on total metal content were considered. As per guidelines of Kabata and Pendias (1984), 3-8 mg total Cd kg -1 soil is considered to be the critical limit above which toxicity of Cd is possible. In the present study, 6 (representing the sites Saleem tabri and Partapsinghwala) out of 53 soil samples reached this threshold value of 3 mg Cd kg-1 soil. ). Thus it appeared that about 11.3 percent soils have become polluted as a result of continuous irrigation with sewage water. These soils require immediate attention and needs to be ameliorated urgently. There is the possibility that rest of the soils might approach this critical limit in a few years if same level of irrigation with sewage water continued. In another standard adopted in U.K, as prescribed by G L.C (Greater London Council), total Cd concentration ranging from 0 to 1, 1 to 3, 3 to 10, 10 to 50 and >50 mg kg-1 soil were categorized as typical uncontaminated, slightly contaminated, contaminated and heavy contaminated soils, respectively. According to this system therefore, most of the investigated soils fell under slightly contaminated category indicating thereby that clean up operation is definitely required. So there is an urgent need to work out the critical limit for Cd toxicity in the Punjab soils on a wider scale 4.2 Influence of different levels of Cd and various amendments on 4.2.1 Dry matter yield of shoots of chalai (pig weed) 4.2.1.1 Effect of Cadmium: Gradual reduction in mean dry matter yield of chalai (pig weed) occurred with increasing levels of cadmium irrespective of the amendments but the significant decrease was observed at and above the application rate of 10 mg kg-1soil. The adverse effect of the added Cd was more marked at highest rate of its application (Table 4.2.1and Fig 2). Mean dry matter yield of chalai in soil at 45 days of growth, significantly decreased from 21.7 g pot-1 to 18.0, 15.4 and 11.5 g pot-1 when the rate of cadmium was increased to 10, 20 and 40 mg kg-1soil irrespective application of amendments. The reduction in dry matter yield of chalai shoots was 1.1, 2.3 16.8, 28.9 and 46.9 percent with the application of cadmium at the rate of 2.5, 5, 10, 20 and 40 mg kg -1soil. The reduction in shoot growth was in accord with the amounts of extractable Cd in the soils and Cd content in plants (Tables 4.2.5 and 4.2.3). The detrimental effect of Cd on yield was consistent with the results of others in different crops (Singh and Nayyar 1991; Dahiya et al 1991; Narwal et al 1990; Cieslinski et al 1996; Bipasha et al 1997; Gupta and Gupta 1988, Khurana et al 2006 and 39

Sidhu and Khurana 2010). This was probably due to marked and significant increase in available Cd which resulted in the increased amount of metal being absorbed by the plants. Atrexe et al (2002) have elucidated that normal growth and development was impaired with Cd which is readily taken up by plants. Cadmium, even at very low concentration results in respiratory and photosynthesis (Lee et al 1976) and structural disorders (Lamoreaux and Chaney 1977). In pea, a number of toxic effects of Cd on metabolism have been reported, such as inhibition of various enzymes activities (Obata et al 1996, and induction of oxidative stress (Sandalio et al 2001) including alterations in enzymes of antioxidant defense system (Romero Puertess et al 2002). The basic cause of Cd toxicity in plants probably lies in much higher affinity of Cd for thiol groupings (-SH) in enzymes and other proteins. Reduced plant growth due to Cd addition might also be attributed to reduced photosynthetic rate and internal water deficit in vascular system caused by reduced conductivity of stem and poor root system development (Alloway 1990). Despite the marked reduction in dry matter yield beyond application rate of 10 mg Cd kg -1 soil, the pig weed plants did not exhibit any visual symptoms of Cd toxicity. Therefore, it may be hypothesized that reduction in dry matter yield had resulted solely from the toxicity of the cadmium and not of nutrient imbalance as no visual symptoms were observed. 4.2.1.2 Effect of amendments: There was significant effect of different amendments on dry matter yield of shoots of pig weed. Different amendments behaved variably as far as their remediation potential was concerned at a particular level of Cd application. Application of various amendments viz (CaCO3, FYM and P) at all the levels mitigated the toxicity of cadmium as is evident by the increase in dry matter yield of the crop irrespective of Cd levels. A significant enhancement in dry matter yield of chalai was observed with all the amendments. Lime @ 2.5 and 5% increased mean dry matter yield from 16.4 (Control) to 18.2 and 19.1 g pot-1, respectively. FYM at the rate 1 and 2% produced mean dry matter yield of 18.4 and 19.4 g pot-1 while 17.6 and 18.4 g pot-1 respectively with phosphorus (P2O5) application of 20 and 40 mg kg-1 soil regardless of Cd levels. Increase in yield may be attributed to alleviating Cd toxicity in soil owing to amendments application (Bolan et al 2003a & b) Our findings are conformity with those of Chen et al (2000); Friesel et al (2004) and Zhu et al (2004) who also reported that application of lime and phosphate amendments significantly increased dry matter yield of fescue grass (Fescue rubra L.) and wheat (Triticum aestivum L.) compared to un-amended control. Among the various amendments, application of CaCO3proved very effective followed by phosphorus (P2O5) and FYM in alleviating the

40

4.2.1: Effect of rates of Cadmium and amendments on dry matter yield of shoots of pig weed.
Cd rates (mg kg-1 soil) 0 2.5 5 10 20 40 Mean Amendment

CaCO3 (%)
Control 20.68 20.09 19.14 15.81 13.27 9.38 16.39 2.50 21.62 21.25 21.36 17.69 15.22 12.00 5 21.92 22.31 21.95 19.15 16.93 12.28

FYM (%)
FYM 1 21.89 22.12 21.48 18.26 15.38 11.52 FYM 2 22.43 22.53 22.22 19.53 17.07 12.40 19.36 = 0.75 = 0.81 = NS

Phosphorus (P2O5 mg kg-1 soil)

20 21.35 20.84 21.02 16.93 14.45 11.20 17.63 40 21.88 20.99 21.14 18.85 15.54 11.75 18.36 Mean 21.68 21.45 21.19 18.03 15.41 11.50

CD(p=0.05)

18.19 19.09 18.44 Cadmium levels Amendments Cadmium X Amendments

Fig 2: Effect of rates of Cadmium and amendments on dry matter yield of shoots of pig weed. Cd toxicity in soil as Cd concentration in shoots decreased maximum with application rate of calcium carbonate (2.5 and 5 % ) and intermediate with phosphorus (20 and 40 mg P2O5 kg-1 soil) and minimum with FYM (1 and 2%). Brown et al (1998) reported that Cd in the sludge amended plot was less available to lettuce than the Cd in the unsludged plot. 41

Maximum mean dry matter yield of the shoots of pig weed was obtained in the treatments involving FYM amendments in spite of the fact it was less effective in decreasing the concentration of Cd in pig weed shoot compared to other amendments. It might be possible that some other useful constituents of FYM were effective in enhancing the yield. The plant appeared healthier in all the FYM treated pots throughout the growing season. Similar effects of compost in growth media has been found in other studies (Atiyeh 2001; Perez Murcia et al 2006) which were mainly due to the improvement of soil structure by increasing porosity, water holding capacity and aeration. Ram and Verloo (1985) also observed that addition of different organic material like FYM, peat, humic acid and tetra ethylene penta amine (Tetren) to a polluted Belgian soil enhanced the dry matter yield of corn, resulting in reduced availability of heavy metals due to formation of insoluble metalloorganic complexes. Clemente et al (2007) found that FYM decreased metal availability in contaminated soils due to formation of insoluble complexes during mineralization of organic matter. Gupta et al (1989) reported that straw and grain yield of wheat grown in highly contaminated soil improved with the increasing levels of FYM in the soils. McBride (1995) observed that substantial fraction of the applied organic matter remains and the elevated organic matter content of soil could limit the activity and bio availability of some heavy metal The increase in shoot yield due to phosphorus application indicated its usefulness and effectiveness to reduce Cd toxicity. Bolan et al (2003) observed that P application reduced phytotoxicity of Cd, resulting in higher yields of Brassica juncea. An enhanced biomass production of Sudex and Swiss chard was reported by Hettiarachhi and Pierzynski (2002) in an experiment conducted on heavy metal pollted soils (Cd, Pb and Zn) with application of soluble source of phosphorus. Basta et al (2001) and Hettiarachchi and Pierzynski (2002) reported that plant tissue concentration of Cd was consistently reduced in the presence of soluble P, possibly through the formation of mixed metal phosphates, which could have restricted metal uptake by plants. The result of present study indicated that in situ immobilization of Cd occurred probably due to the phosphate induced Cd adsorption and precipitation of Cd as Cd (OH)2 and Cd3(PO4)2 with the addition of mono calcium carbonate. This corroborated result of (Bolan et al 2003). Amendment in the form of CaCO3 was found to be most effective. It was probably due to formation of less soluble compounds like CdCO3 (Bolan et al., 2003a) mitigated the toxicity of cadmium. An other explanation may be that increased levels of Ca2+ through the application of calcium carbonate can decrease the amount of Cd that is assimilated by plants (Larlson et al 2000) because of their similar size (Ochiai 1995). A higher affinity for the essential trace metal Ca results in the decreased uptake of Cd into the plant. These findings find support from the work of Singh and Nayyar (2001) who reported that 2.5 percent application of CaCO to the soil resulted in the decrease in both DTPA-extractable and plant Ni 42

which helped in mitigating the toxic effect of Ni and subsequently increased dry matter yield of cowpea. Peles et al (1998) concluded that the addition of lime to contaminated soils (essentially increasing the pH) decreased the uptake of heavy metals including cadmium. Lee et al (2004) and Zhu et al (2004) reported that lime and phosphorus significantly decreased the concentration of heavy metals including Cd in wheat shoots. Ram and Verloo (1985) found that FYM and peat soil enhanced the mobility of Cd at lower pH and decreased it at higher pH. 4.2.2 Dry matter yield of roots as influenced by Cd and amendments application 4.2.2.1 Effect of Cadmium: Roots dry matter yield of pig weed followed the same trend as that case of shoot. The dry matter yield of roots started declining even with the application rate of 5 mg Cd kg-1 soil. However, a significant decrease was observed with 10 mg Cd kg-1 soil onwards. Further, application of 20 and 40 mg Cd kg-1 soil showed significant sharp decline in root dry matter yield. This indicated that root yield recorded by the addition of 20 and 40 mg Cd kg-1 soil was significantly lower than the root yield recorded with 10 mg Cd kg 1

soil. The reduction in dry matter yield of pigweeds root was 16.0, 28.2 and 45.9 percent with

the application of cadmium at the rate of 10, 20 and 40 mg kg-1soil. So the yield reduction in response to Cd was of similar proportions in the roots as for the aboveground part of the plants. The reduction in root growth was in accord with the amounts of extractable Cd .The magnitude of reduction in root biomass was comparable to the shoot biomass (Tables 4.2.2 & 4.2.1and Fig 2 & 3) 4.2.2.2 Effect of Amendments: The effect of different amendments on root growth was similar to that reported for the above ground part of the plants. Different amendments helped to ameliorate the toxicity of Cd to varying extent and subsequently enhanced the dry matter yield. As in case of shoot, all the amendments registered significant increase in dry matter yield of the crop A significant enhancement in dry matter yield of roots of chalai was observed from 5.4 to 5.9, 6.2; 6.1, 6.3; 5.8 and 6.0, respectively with application rate of calcium carbonate (2.5 and 5 % percent), FYM (1 and 2%) and phosphorus application (20 and 40 mg P2O5 kg-1 soil) regardless of Cd levels. Calcium carbonate amendment was proved to be most effective among all the treatments as it offset the toxic effect of cadmium more efficiently. Similarly maximum dry biomasses of the roots were obtained with FYM amendments as in case of shoots. Same reasoning can be put forth for the behavior of different amendments as case of the roots.

43

Table 4.2.2: Effect of rates of Cadmium and amendments on dry matter yield of roots of pig weed Cd rates (mg kg-1 soil) 0 2.5 5 10 20 40 Mean Amendments CaCO3 (%) Control 6.82 6.58 6.38 5.26 4.42 2.88 5.40 2.5 6.94 7.0 7.00 5.80 4.90 3.90 5 6.98 7.24 7.14 6.20 5.50 4.00 1 7.12 7.26 6.98 6.03 5.04 4.10 FYM (%) 2 7.22 7.34 7.26 6.32 5.62 4.12 6.32 = 0.33 = 0.36 = NS Phosphorus (P2O5 mg kg-1 soil) 20 7.04 6.81 6.78 5.54 4.74 3.78 5.78 40 7.0 6.88 6.92 6.18 5.08 3.86 5.98 7.02 6.96 6.92 5.90 5.04 3.80 Mean

CD(p=0.05)

5.92 6.18 6.08 Cd levels Amendments Cd levels X Amendments

Fig 3: Effect of rates of Cadmium and amendments on dry matter yield of roots of pig weed

44

4.2.3 Effect of Cd and amendments levels on Cd content (gg-1) in shoots and roots of pig weed 4.2.3.1 Effect of Cadmium: The mean concentration of Cd in the shoots increased successively with increasing rates of Cd application irrespective of applied amendments. The application of even the lowest rate of 2.5 mg Cd kg-1 soil significantly increased the Cd content in crop over control. The mean Cd concentration increased from 1.3 in control treatment to 4.7, 6.0, 14.1, 21.8 and 46.4 g g-1 dry matter when rate of Cd application was raised to 2.5, 5, 10, 20 and 40 mg kg-1 soil. The difference of Cd content among the treatments were also significant. This was probably due to marked and significant increase in available Cd (Table 4.2.3 and Fig 4) which resulted in the increased amount of metal being absorbed by the plants. Compared to its value in shoot, contents of cadmium in roots were invariably higher at all levels of added cadmium i.e consistently higher concentration of Cd was detected in roots as compared with shoots (Table 4.2.3 and Fig 5). Since roots are the primary plant organs which remain in contact with soil solution and thus accumulate comparatively higher amount of heavy metals. An application rate of 5 mg Cd kg-1 soil was required to significantly increase the root Cd content in pigweed over control regardless of amendments levels. It is pertinent to note here that although this level significantly increased the Cd content in both shoots and roots but they failed to cause a significant reduction in the dry matter yield of the crops, indicating that Cd is less phyto-toxic at these level. The highest Cd concentration was obtained with 40 mg Cd kg-1 soil both in shoots and roots. These results find support from the work of several workers namely, Maclean (1976); Mahler et al (1978); Singh and Nayyar (1989); Narwal et al (1993); Khurana et al (2006) and Sidhu and Khurana (2010) who also reported higher Cd content in different crops due to Cd application. 4.2.3.2 Effect of Amendments: Cadmium concentration both in roots and shoots showed decreasing trend with supply of amendments. However, there was considerable difference in magnitude of different amendments in alleviating Cd toxicity by bringing decrease in Cd concentration in both shoots and roots of the crop. All the amendments significantly decreased the concentration of Cd in shoot and roots of pigweed (Table 4.2.3 and Fig 4 & 5) The concentration of Cd in shoot with lime @ 2.5% and 5% treated soils decreased from 20.7 to 14.2 and 12.8 g g-1 dry matter which was 31.5 and 37.8% lower than that in the control plants, respectively irrespective of Cd levels. FYM addition @ 1 and 2% reduced concentration of Cd in shoot from 20.7 (control) to 16.8 and 15.3 which comes out to be 18.8% and 26.2% lower that in control. Addition of

45

phosphorus @ 20 and 40 mg P2O5 kg-1 soil decreased shoot Cd concentration by 22.7 and 30.2%, respectively compared to that in the control plants. Table 4.2.3: Effect of rates of Cadmium and amendments on concentration of Cadmium in shoots and roots of pig weed. Amendments Shoots Cd rates (mg kg-1 soil) 0 2.5 5 10 20 40 Mean 0 2.5 5 10 20 40 Mean CaCO3 (%) Control 1.60 6.04 8.06 18.66 29.06 60.58 20.66 2.18 9.28 13.22 24.80 39.38 79.34 28.04 2.50 1.20 4.18 5.72 12.40 20.08 41.30 14.16 1.60 5.20 7.02 16.78 27.24 50.04 17.98 5 1.02 3.64 4.52 11.44 16.76 39.77 12.86 1.42 4.48 5.98 14.26 22.12 46.90 15.86 FYM (%) 1 1.28 5.02 6.50 15.38 24.10 48.36 16.76 Roots 1.66 7.36 10.18 19.74 31.02 51.98 20.32 1.54 6.30 9.00 17.72 26.66 51.00 18.72 1.58 6.28 8.94 19.06 30.40 54.24 20.08 = 0.69 = 0.74 = 1.82 = 0.86 = 0.93 = 2.28 1.60 5.74 8.10 16.22 26.75 52.10 18.42 1.66 6.38 8.92 18.38 29.08 55.10 2 1.20 4.64 5.88 13.82 21.16 44.80 15.26 Phosphorus (P2O5 mg kg-1 soil) 20 1.32 4.88 6.06 14.68 22.34 46.52 15.98 40 1.28 4.64 5.50 12.34 19.10 43.66 14.42 Mean 1.26 4.72 6.04 14.10 21.80 46.42

CD(p=0.05)

Cadmium levels (shoots) Amendments (shoots) Cadmium X Amendments (shoots) Cadmium levels (roots) Amendments (roots) Cadmium X Amendments (roots)

CD(p=0.05)

46

Fig 4: Effect of rates of Cadmium and amendments on concentration of Cadmium in shoots of pig weed

47

Fig 5: Effect of rates of Cadmium and amendments on concentration of Cadmium in roots of pig weed Similarly reduction in mean Cd content due to application of various amendments was registered in roots as for the above ground part of the plants. The amendments had significant effect in decreasing the concentration of Cd in roots compared to that in the control plants. Lime @ 2.5% and 5% decreased root Cd concentration by 35.9 and 43.4 compared to that in control samples. FYM addition @ 1 and 2% declined it by 27.5% and 33.2% compared to that in the control samples. The mean concentration of Cd in root samples in the presence of 20 and 40 mg P2O5 kg-1 soil was 28.4 and 34.3 % lower than that in the control, respectively. The decrease in concentration may be due to growth dilution, which occurred with an increase in biomass production since there was an increase in yield owing to application of lime, P2O5 and FYM (Lee et al 2004) and partially decreased Cd concentration in soil solution with all the amendments through formation of less soluble compounds like Cd3(PO4)2 and CdCO3 (Bolan et al 2003a). Lee et al (2004) and Zhu et al (2004) reported that lime, compost and phosphorus significantly decreased the concentration of heavy metals including Cd in wheat grain and straw compared to that of control plants. Similarly a decrease in Cd concentration through P application were obtained by Dheri et al (2007) and Hettiarachchi and Pierzynski (2002) in spinach and swiss chard (Beta vulgaris (L). Clemente et al (2007) and Mc Bride (1995) demonstrated the effectiveness of FYM in decreasing Cd concentration due to formation of insoluble complex. 4.2.4 Effect of Cd and various amendments levels on Cd uptake of shoots and roots 4.2.4.1 Effect of cadmium: The data on the Cd uptake as affected by added Cd and various amendments in the shoots are presented in table 4.2.4. The mean Cd uptake in the shoots of pig weed increased significantly with increasing level of Cd application regardless of Cd application. The Cd uptake in pigweed increased from 27.5 g pot -1 in control to 100.8, 126.8, 251.9, 331.9, 528.2 g pot-1with 2.5, 5, 10, 20 and 40 mg Cd kg-1 in soil respectively. Compared with no Cd (control) a 9.7 fold increase in its uptake was observed by applying 10 mg Cd kg-1. It suggested that applied Cd was readily absorbed by crop and was easily translocated from roots to above ground plant parts. Raising the Cd level to 20 mg kg-1 soil did not increase the Cd uptake to an extent similar to that observed with 10 mg Cd kg -1 soil. In fact applying 20 mg Cd kg-1 soil, only 1.3 times increase in Cd uptake was observed over that recorded at 10 mg Cd kg -1 soil. This kind of Cd uptake pattern was consequence of both reduction in yield due to Cd toxicity and increased uptake of Cd at higher application rate as a reduction in yield was compensated by higher Cd absorption. Lower uptake of cadmium in roots of pig weed as compared to shoots, inspite of its high Cd concentration had resulted from the lower dry matter yield of the roots (Table 4.2.2 48

& 4.2.3). The mean uptake of Cd by roots increased significantly and progressively with increasing levels of Cd (Table 4.2.4 and Fig 6 & 7). The increase in mean uptake of Cd in roots of pigweed was about 3.9, 5.3, 9.3, 12.5 and 17.8 times higher than control at 2.5, 5, 10, 20 and 40 mg Cd kg-1 soil application 4.2.4.2 Effect of Amendments: Application of different amendments decreased Cd uptake in both shoots and roots of pig weed significantly. As explained earlier, application of various amendments had depressed the availability of Cd (Table 4.2.3) and increased the dry matter yield, thereby a reduced Cd uptake in pigweed plant was observed. The uptake of Table 4.2.4: Effect of rates of Cadmium and amendments on uptake of Cadmium by shoots and roots of pig weed. Amendments Shoots Cd rates (mg kg-1 soil) 0 2.5 5 10 20 40 Mean 0 2.5 5 10 20 40 Mean CaCO3 (%) Control 32.3 120.9 154.0 294.7 383.8 569.4 259.2 14.5 61.1 84.3 130.6 172.7 229.1 115.4 2.50 25.7 88.2 122.1 219.3 305.4 496.1 209.5 11.2 36.2 49.0 96.8 133.4 195.2 87.0 5 22.8 80.8 98.7 218.8 284.2 487.1 198.7 10.0 32.5 42.6 88.8 121.9 186.5 80.4 FYM (%) 1 27.9 111.4 138.7 280.5 370.6 556.1 247.5 Roots 11.8 53.8 71.2 118.4 156.5 212.9 104.1 11.2 46.4 65.6 111.4 150.3 210.0 99.1 11.2 42.8 60.5 105.9 144.0 204.5 94.8 11.1 39.4 55.7 99.6 135.7 202.0 90.6 11.6 44.6 61.3 107. 3 144. 9 205. 7 2 27.3 105.2 130.5 269.4 360.4 555.3 241.4 Phosphorus (P2O5 mg kg-1 soil) 20 28.2 101.8 127.1 248.9 322.0 521.1 224.8 40 28.0 97.5 116.6 231.4 296.9 512.4 213.8 Mean 27.5 100.8 126.8 251.9 331.9 528.2

49

CD(p=0.05)

Cadmium levels (shoots) Amendments (shoots) Cadmium X Amendments (shoots) Cadmium levels (roots) Amendments (roots) Cadmium X Amendments (roots)

= 13.58 = 14.67 = NS = 9.19 = 9.93 = NS

CD(p=0.05)

Fig 6: Effect of rates of Cadmium and amendments on uptake of Cadmium by shoots of pig weed

50

Fig 7: Effect of rates of Cadmium and amendments on uptake of Cadmium by roots of pig weed Cd in shoot decreased from 259.2 g pot-1 (control) to 209.48 (lime @ 2.5%), 198.7 (lime @ 5%); 247.5 ( FYM @ 1% ), 241.4 (FYM @ 2% ); 224.8 (P @ 20 mg P2O5 kg-1 soil) and 213.8 g pot-1 (P @ 40 mg P2O5 kg-1 soil). Application of lime @ 2.5% and 5% decreased it by 19.2 and 23.3 % lower compared to control plants, respectively irrespective of Cd levels. FYM addition @ 1 and 2% reduced uptake of Cd in shoot by only 4.5 and 6.9 percent lower that in control. Addition of phosphorus @ 20 and 40 mg P2O5 kg-1 soil decreased shoot Cd concentration by 13.3 and 17.5 per cent respectively compared to that in control. Application of various amendments viz (FYM, CaCO3, and Phosphorus) at all the levels reduced the uptake of cadmium which reflected in the increase in dry matter yield of the crop irrespective of Cd levels (Tables 4.2.1 and 4.2.2). A significant reduction in uptake of cadmium was observed with all the amendments. Lime @ 2.5 and 5% decreased it by 28.4 and 35 g pot-1 respectively with respect to control. FYM at the rate 1 and 2% declined it by 11.3 and 16.2 g pot-1 while 20.5 and 24.8 g pot-1 reduction in uptake of the crop respectively was obtained with phosphorus application of 20 and 40 mg P2O5 kg-1 soil) with respect to control regardless of Cd levels. Singh et al (1991) observed that when Cd was applied with FYM, the uptake of Cd decreased with increasing level of FYM. Similar decrease in Cd uptake with increasing application of FYM had also been reported by Dahiya et al (1987). The decrease in DTPA extractable Cd (Table 4.2.5) in soils with calcium carbonate may be result of directly through stronger binding and precipitation of this metal with CaCO3 and indirectly rise in pH of the soils (Singh and Nayyer 1993). Liming can lead to the precipitation of metals as metal-carbonate and significantly decrease the exchangeable fraction (Table 4.4b) of metals in contaminated soil (Knox et al 2001) which could reduce the uptake by plants. Ma and Uren (1998) observed marked decrease in DTPA extractable with addition of CaCO3 through its effect on increase in pH Cd contents in wheat after addition of lime to soil were significantly decreased and proposed that CaCO 3 can help in mitigating the toxic effects of Cd on wheat. 4.2.5 DTPA extractable Cd before sowing of crops: Effect of Cd on DTPA extractable Cd in soils after equilibrium is presented in table (4.2.5 & Fig 8). The data showed that extractable Cd increased markedly and significantly with graded rates of Cd application irrespective of amendments before sowing of crop. The increase was significant with 5 mg Cd kg-1 soil application over control. The content of DTPACd in soil after completion of equilibrium time were 0.27, 0.65, 1.68, 3.18, 7.37, and 14.91 mg kg-1 soil at 0, 2.5, 5, 10, 20 and 40 mg Cd kg-1 soil application. A perusal of data in table (4.2.5) showed that the maximum extraction of Cd by DTPA with 40 mg Cd kg-1 soil application was approximately 51

60.4 percent in the absence of amendments which was less than 70 to 80 percent of applied Cd that reported by Haq et al (1980). Singh and Nayyar (1989) also obtained about 60 to 80 percent of added Cd in coarse textured soil. Cieslinski et al (1996) reported 13.7, 26.7, 51.9 mg Cd kg-1 soil DTPA extractable Cd where the application rates were 15, 30 and 60 mg kg-1 soil. This showed high rate of cadmium extractability, which is a cause of concern.

Table 4.2.5: Effect of rates of cadmium and amendments on DTPA-Cd in soil at equilibrium Amendments Cd rates (mg kg-1 soil) 0.00 2.50 5.00 10.00 20.00 40.00 Mean CD(p=0.05) CaCO3 (%) Control 0.36 1.13 2.34 5.27 11.02 24.17 7.38 2.5 0.20 0.48 1.34 2.44 6.08 12.21 5 0.15 0.32 1.22 2.02 5.28 11.82 FYM (%) 1 0.31 1.02 1.92 3.43 8.08 14.86 4.94 2 0.30 0.64 1.81 3.22 7.62 14.30 Phosphorus (P2O5 mg kg-1 soil) 20 0.28 0.60 1.66 3.06 7.37 13.92 4.41 40 0.25 0.56 1.48 2.75 6.14 13.08 4.05 Mean 0.27 0.65 1.68 3.18 7.37 14.91

3.80 3.50 Cadmium levels Amendments Cadmium X Amendments

4.64 = 0.71 = 0 .76 = 1.87

52

Fig 8: Effect of rates of cadmium and amendments on DTPA-Cd in soil at equilibrium 4.2.5.1 Effect of amendments: Applied amendments were found to be significantly

effective in declining the content of DTPACd in soil over control after the completion of equilibration period. The content of DTPA-Cd decreased by 3.58 mg kg-1 soil (lime @ 2.5%), 3.88 (lime @ 5%); 2.44 ( FYM @ 1% ), 2.74 (FYM @ 2% ); 2.97 (P @ 20 mg P2O5 kg-1 soil) and 3.33 g pot-1 (P @ 40 mg P2O5 kg-1 soil) as compared to control. Corresponding percent decrease comes out to be 48.5, 52.6, 33.1, 37.1, 40.2 and 45.1 respectively with application rate of calcium carbonate (2.5 and 5 % percent), FYM (1 and 2%) and phosphorus application (20 and 40 mg P2O5 kg-1 soil) regardless of Cd levels respectively. The increase in soil pH with lime (Table 4.5.1) might cause in precipitation of Cd with carbonates and reduced the solubility of Cd in soil. Decrease phosphorus may be caused by significant increase in Cd application significant increase in Cd
2+ 2+

in DTPA-Cd with

adsorption. Similar reasoning has

been put forth by Naidu et al. (1996) and Bolan et al. (1999) that increasing rate of phosphate adsorption by soil colloids. Therefore, decrease in DTPA extractable Cd may be attributed to an increase in negative charge to affect more Cd adsorption. The precipitation of Cd as Cd(OH)2 and Cd(PO4)2 has also been reported by Bolan et al. (2003a) with the application of P amendments. Formation of insoluble oregano metallic complexes may be the reason behind the reduction of DTPA-Cd in soil with FYM application (Clemente et al 2007). 4.2.6 DTPA extractable cadmium in post harvest soils: Compared to its content in equilibrated soil (table 4.2.5), the DTPA Cd in general decreased in soil after the harvest (Table 4.2.6). Perusal of data in table (4.2.6) revealed lower values of DTPA-Cd in all the treatments after the harvest of the crop compared to its value at equilibrium. The contents of Table 4.2.6: Effect of rates of Cadmium and amendments on DTPA-Cd in soil after crop harvest Cd rates (mg kg-1 soil) 0 2.5 5 10 20 40 Mean Amendments CaCO3 (%) Control 0.20 0.63 1.98 3.86 9.24 19.81 5.95 2.5 0.12 0.29 1.14 1.88 5.18 9.95 3.09 5 0.07 0.26 1.08 1.62 4.59 9.52 2.85 1 0.13 0.62 1.70 2.60 6.72 12.54 4.05 53 FYM (%) 2 0.13 0.46 1.55 2.35 6.30 11.66 3.74 Phosphorus (P2O5 mg kg-1 soil) 20 0.16 0.38 1.40 2.26 6.09 11.54 3.64 40 0.14 0.33 1.31 2.09 5.41 10.80 3.35 Mean 0.14 0.42 1.45 2.38 6.22 12.26

CD(p=0.05)

Cadmium levels Amendments Cadmium X Amendments

= 0.65 = 0.68 = 1.69

Fig 9: Effect of rates of Cadmium and amendments on DTPA-Cd in soil after crop
harvest DTPACd in soil after crop harvest were 0.14, 0.42 , 1.45 , 2.38 6.22, 12.26 mg kg-1 soil as against 0.27, 0.65, 1.68, 3.18, 7.37, and 14.91 mg kg-1 soil at 0,2.5, 5, 10, 20 and 40 mg kg-1 soil application at equilibrium. This might be due to removal of Cd by the crop as well as transformations in to relatively insoluble forms. The minimum and maximum amount of DTPA Cd left after the harvest was 2.85 and 4.05 mg kg-1soil with application of 5 percent calcium carbonate and 1 per cent FYM respectively. It was apparent from the data in table (4.2.6 & Fig 9) that the amount of DTPA left in soil after the crop harvest in the presence of amendments was significantly lower as compared in their absence. 4.3: Effect of Cd levels on micronutrient contents (g g1 dry matter) in pig weed: Plant samples were collected from the treatments involving various levels of Cd in order to study the effect of these levels on concentrations of micronutrients (Fe, Zn, Cu and Mn) Table 4.3.1: Effect of Cd on the Fe content (g g-1 dry matter) in the shoots and roots of pigweed Cd rates 54 Pigweed

Shoots 0 2.5 5 10 20 40 CD(p=0.05)

313.3 322.9 330.9 349.9

Roots
396.4 405.8 417.0 441.5 427.6 420.2

338.8 324.2 9.9

11.0

4.3.1 Iron
The iron concentration in the roots and shoots of pig weed revealed a synergistic relationship between Cd and Fe up to 10 kg levels (20 and 40 mg Cd kg
-1 1 1

soil and an antagonistic relationship at higher

-1

soil). It showed increasing trend in shoot from 313.34 in soil and This is

control to 349.94 gg when the levels of Cd were raised to 10 mg kg

further supported by the high positive correlation between Cd and Fe in the shoots (r = 0.91 Cd kg 1 soil) and roots (r = 0.93). The mean Fe concentration in the shoots and roots showed a decreasing trend beyond an application rate of 20 mg Cd kg 1 soil. Negative correlation (r = -0.80 in shoot and -0.65 in roots) further lend support to These findings are in conformity with the results of Khan and Khan (1983); Koshino (1973); Root et al (1975) and Rupp et al (1985), who reported similar behaviour in tomato and egg plant, rice, corn and grapevine, respectively. Gupta and Dixit (1992), on the other hand, reported that the Fe content decreased due to the application of Cd in soybean and wheat, while Mahler et al (1982) found no influence of Cd application on the leaf Fe concentration of lettuce or Swiss chard. Table 4.3.2: Effect of Cd on the Cu content (g g-1 dry matter) in the shoots and roots of pigweed Cd rates Shoots 0 2.5 5 10 20 40 CD(p=0.05) 4.3.2 Copper 12.4 11.6 10.7 8.7 8.0 7.7 1.4 Pigweed Roots 14.7 13.9 12.3 10.6 9.7 9.2 1.9

55

The copper concentration in the roots and shoots of pig weed was negatively affected by the application of Cd at all levels (Table 4.3.2). There was a gradual decrease in Cu concentration in the shoots and roots of the crop when the levels of cadmium were raised from 0 to 40 mg kg-1soil. It decreased from 12.35 g g-1 to 11.55, 10.7, 8.75, 8.05 and 7.65 g g-1 when the levels of Cd were raised to 5, 10, 20, 40 and 80 mg kg -1 soil. The correlations between these metals were found to be highly negative both in the shoots (r = 0.77) and in the roots (r = 0.75). This may be ascribed to antagonism between the two elements. The copper content in the roots of the crops was generally higher than in the shoots. These findings are in conformity with the results of Khan and Khan (1983). The antagonistic effect of Cd accumulation on the levels of essential nutrients in the leaves was also observed in chelator-buffered nutrient solution (Adhikari et al 2006). 4.3.3 Zinc The data in table 4.3.3 revealed consistent increase in the mean Zn concentration with increasing levels of Cd up to 10 mg kg 1 soil, while the higher rates (20 and 40 mg kg 1 soil) depressed the zinc concentration in both the roots and shoots of the crops. This showed that low levels of cadmium have a synergistic effect on the zinc concentration in the shoots and roots while high levels have an antagonistic effect. A significant positive correlation was observed between the rate of Cd application and the Zn concentration of the shoots (r = 0.90) and roots (r=.89) up to the 10 mg Cd kg1 soil rate. At higher rates, however, highly significant negative correlations were obtained for both the shoots (r = 0.92) and the roots (r = 0.94). The increase in shoot zinc concentration with the application of Cd up to 10 mg kg
1

soil may be due to the concentration effect, because the dry matter yields of the shoots and

roots were decreased by cadmium. Other possible explanation for this apparent synergism was that Cd dissociated zinc fixed from the binding sites in the soil due to competition for the same sites. Table 4.3.3: Effect of Cd on the Zn content (g g-1 dry matter) in the shoots and roots of pigweed Cd rates Shoots 0 2.5 5 10 20 40 CD(p=0.05) 52.5 56.6 58.4 62.8 60.1 50.9 2.14 Pigweed Roots 69.6 73.9 77.8 83.2 78.6 66.8 3.70

56

Other workers (MacLean 1976; Singh and Steinnes 1976 and White and Chaney 1980) did not report any interaction between Cd and Zn in lettuce, barley and soybean respectively. However Abdel-Sabour et al (1988) observed decreased Cd content with Zn application in maize. Oliver et al (1994) investigated the accumulation of cadmium by wheat grown at nine sites of South Australia as affected by zinc application and found a marked decrease of Cd concentration in wheat grain. The result clearly indicated their antagonistic relationship But Iwai et al (1975) observed that Zn added to substrate had no effect of Cd content of corn. 4.3.4 Manganese There was a consistent decrease in the mean Mn concentration of the shoots with increasing levels of cadmium, which could be interpreted as a clear cut case of antagonism between Cd and Mn (Table 4.3.4). The application of 5,10, 20 and 40 mg Cd kg-1soil decreased significantly mean Mn concentration in the shoots crop by 5.7, 14.1, 27.1 and 31.6 percent respectively as compared to control Similarly In the roots, the application of 5 mg Cd kg1 soil significantly decreased the Mn concentration. These findings were corroborated by the negative correlation detected in both the shoots (r = 0.89) and the roots (r = 0.87). Cataldo et al. (1983) found that pearl millet and green gram showed a depression in the Mn Table 4.3.4: Effect of Cd on the Mn content (g g-1 dry matter) in the shoots and roots of pigweed Cd rates Shoots 0 2.5 5 10 20 40 CD(p=0.05) 73.2 71.9 68.9 62.8 53.4 50.1 4.5 Pigweed Roots 81.3 80.2 76.3 69.2 58.9 55.4 4.5

content when 10 and 20 mg Cd kg1 soil was added. These authors attributed this to the fact that Cd competitively inhibited Mn absorption, suggesting a common transport site or process. Patel et al (1976) also observed that the Mn concentration decreased due to Cd application. On the other hand, Khan and Khan (1983) and Narwal et al (1993) reported that the Mn concentration increased with the addition of cadmium, in contrast to the findings in the present study 4.4) Upper critical level of Cd in soil and plant 57

4.4.1 4.4.2

Soil Plant

4.4.1 Toxic level of Cd in soil: The toxic or upper critical level of Cd is defined as its lowest concentration in tissue or soil at which its presence led to reduction in yield. The critical concentration can be calculated by interpolating the concentration at which yield is reduced by some arbitrary amount usually 10 to 30 percent. For finding the upper critical level, the regression equations were worked out where the value of percent reduction in dry matter yield was regressed with corresponding DTPA- Cd in soil. Different mathematical and regression models such as linear, quadratic and exponential were tested using MS excel (Windows 2007) to find out the best fit for establishing the critical level . There was excellent fit to the data

with

quadratic model as revealed by significant coefficient of determination (R2)

From these equations, the toxic level of DTPA Cd at which 20 percent reduction in dry matter yield occurred, were then estimated. The toxic levels of DTPA Cd was found to be 4.38mg kg-1 soil for pig weed Table 4.4: Upper critical levels of cadmium in soil and plant in pig weed Equation Soil Linear Quadratic Exponential y = - 1.1053+0.4084 X1 y= 0.9168+ 0.0234 X1 +0.0073 X12 y= 0.8035e0.0681x1 Plant Y1=0.039+.0.995X1 Y1=3.91+0.253X1+0.014X12 Y1=3.678e0.0558x1 0.91 0.93 0.84 Coefficient of determination (R2)

Linear Quadratic Exponential

0.92 0.90 0.86

4.4.2 Toxic levels of Cd in shoot of pig weed:. Same method as used for shoot was employed for finding the toxic level of Cd in soil for a crop through regression equations where the values of percent reduction with Cd levels in dry matter yield of shoot of crop were regressed with corresponding tissue Cd concentration. The toxic levels of Cd were then estimated from the equations at 20 percent reduction. Here also, excellent fit to the data

was found with quadratic model as indicated by significant coefficient of determination (R2)The toxic levels of Cd in shoots of pig weed at grand growth stage
through quadratic model was found to be 14.6g g-1 dry matter respectively. 58

4.5 Laboratory Studies: The soil samples treated with various levels of Cd and amendments (Calcium Carbonate @ 5%, FYM @ 2% and Phosphorus @ 40mg kg-1 soil) were collected after pooling the replications treatment wise at equilibrium and after the harvest of the each crop. Physico chemical characteristics of the soil samples (24 at equilibrium and harvest) such as pH, organic carbon and CaCO3 pertaining to above treatments were determined and are presented in table 4.5a. These soil samples (24 at equilibrium and harvest) were then subjected to sequential fractionation to determine fractions such as Exchangeable + water soluble (EX+WS), carbonate bound (CARB), Organic bound fraction (OM), Mn oxide bound (MnOX), Amorphous Fe oxide bound (A FeOX), crystalline Fe-oxide bound (C FeOX) and Residual (RES). Influence of various amendments was studied on the distribution and transformations of cadmium and results are discussed as under: Table No 4.5.1: Physico chemical characteristics of the soil as influenced application of with Calcium carbonate, FYM and Phosphorus at equilibrium Treatments pH Organic carbon(%) CaCO3 (%)

CaCO3 FYM Phosphorus Control Soil

5.0 % 2.0 % 40 mg kg-1

7.92 7.10 7.00 7.02

0 .20 0 .65 0.24 0.24

4.62 0.16 0.18 0 .18

4.5.1 Effect of amendments on Physico chemical characteristics of the soil: Soil pH, calcium carbonate, organic matter: Among factors controlling the concentration of Cd in soil, pH, CaCO3 and organic matter are probably among the important parameters (Christensenn 1984; Ghafoor et al 2008). Addition of various amendments viz calcium carbonate @ 5 % percent, FYM @ 2% and phosphorus @ 40mg kg-1 soil altered original properties of the soil. Soil pH increased markedly in soils treated with calcium carbonate at the rate of 5 percent respectively but to lesser extent with application of other amendments. As expected, addition of calcium carbonate 5 % resulted in increase of CaCO3 content of the soil. The increase in organic content was especially noticeable in soils treated with and FYM @ 2%. This material contained 20.18 percent of organic carbon (Table: 3.3)

59

Table 4.5.2: Distribution of Cd in different fractions as influenced by Cadmium and amendments at equilibrium Control Rates of Cd 0 2.5 5 10 20 40 EX+WSCd 0.06 0.40 0.95 1.85 2.98 4.54 CaCO3-Cd 0.12 0.18 0.38 0.60 1.12 2.05 OM-Cd 0.32 0.39 0.78 1.42 2.84 5.24 CaCO3 (5 %) Rates of Cd 0 2.5 5 10 20 40 EX+WSCd
0.03 0.16 0.32 0.78 1.02 1.54

MnOXCd 0.57 0.60 0.68 0.98 1.65 5.79

A FeOXCd 0.88 1.72 1.38 3.14 5.28 8.74

C FeOXCd 0.57 0.88 1.26 1.35 1.80 4.18

RESCd 1.39 1.69 2.61 4.08 7.62 12.60

CaCO3-Cd
0.26 0.51 0.85 1.99 3.46 6.54

OM-Cd
0.08 0.16 0.43 0.78 1.68 2.88

MnOXCd
0.70 1.18 1.40 1.72 2.04 6.12

A FeOXCd
1.00 1.34 1.46 3.18 5.58 9.02

C FeOX-Cd
0.58 0.94 1.34 1.38 1.88 4.28

RESCd
1.30 1.67 2.35 3.98 7.98 13.10

FYM (2 %) Rates of Cd 0 2.5 5 10 20 EX+WSCd

0.03 0.22 0.44 0.98 1.22

CaCO3-Cd
0.03 0.09 0.16 0.32 0.53

OM-Cd
0.40 0.54 1.52 2.42 5.08

MnOXCd
0.78 1.18 1.32 1.72 2.28

A FeOXCd
1.00 1.32 1.50 3.78 6.02

C FeOXCd
0.59 0.91 1.24 1.44 1.89

RESCd
1.08 1.43 1.86 2.60 6.08

60

40

2.00

1.43

8.12

6.68

9.32

4.48

10.78

Phosphorus (P2O5 40 mgkg-1 soil) Rates of Cd 0 2.5 5 10 20 40 EX+WSCd

0.03 0.20 0.34 0.84

CaCO3-Cd 0.10 0.23 0.38 0.69 1.16 2.11

OM-Cd 0.20 0.52 0.99 1.45 3.04 6.02

MnOXCd
0.68

A FeOXCd 0.98 1.24 1.42 3.22 5.86 9.18

C FeOXCd 0.60 0.92 1.32 1.42 1.88 4.44

RESCd 1.35 1.84 2.48 4.65 8.10 12.9 1

1.02 1.12 1.32 2.28 6.62

1.04 1.80

61

Fig 10: Distribution of Cd in different fractions as influenced by Cadmium and amendments at equilibrium.

62

Distribution of Cd in different fractions as influenced by Cadmium and amendments (at equilibrium and post harvest): 4.5.2: Exchangeable + Water soluble fraction (EX+WS) 4.5.2.1 Effect of cadmium: Exchangeable and water soluble Cd increased with increasing levels of cadmium (Table 4.5.2). It increased from 0.06 to 4.54 mg Cd kg-1 when Cd application was increased from 0 to 40 mg kg-1 at equilibrium. After the crop harvest there is slight decrease (0.04 to 3.94) in this fraction. This might be due to the crop removal or transformation of this pool to the other pools 4.5.2.2 Effect of Amendments: Application of amendments decreased the amount of Cd in this fraction to the varying degree of magnitude at equilibrium and after harvest. At 40 mg Cd kg-1soil, it decreased from 4.54 to 1.54 mg kg-1 soil with application of CaCO3 at the rate of 5percent, to 2.00 mg kg-1 soil with FYM at the rate of 2% and to 1.80 mg kg-1 soil when phosphorus was applied at the rate 40 mg kg-1 soil. Liming decreased the amount of EX+WS fraction due to increase in pH (Table 4.5.1). These observations are in line with those of Chen et al (2000); Bolan et al (2003b; Zwonitzer et al (2003) and Zhu et al (2004). Narwal and Singh (1998) observed that application of cow and pig manure decreased the exchangeable and water soluble cadmium in soils. They argued that formation of organic complexes was more important than CEC in reducing this fraction. Iwegbue et al (2007) suggested that the organic compounds bound more trace metals than held on the exchange sites. 4.5.3: Carbonate bound fraction (CARB-Cd) 4.5.3.1 Effect of cadmium: An increasing trend was noticed in the amount of carbonate bound fraction with increasing rates of Cd application levels (Table 4.5.2) at equilibrium. It increased from 0.12 to 2.05 mg Cd kg-1 soil when Cd application was increased from 0 to 40 mg Cd kg -1soil which amounted to 2.95 and 4.76 percent at the above rates respectively. After the crop harvest there was slight decline (2.60 to 4.27 percent) in this fraction. This may be due to transformation of this fraction into the other fraction with time. 4.5.3.2 Effect of Amendments: The amendments differed to affect carbonate bound Cadmium Maximum amount of applied Cd at all the rate of its application associated itself in this fraction was observed in the treatment having 5% CaCO3 followed by, phosphorus @ 40mg kg-1 and FYM at the rate of 2 percent. At 40 mg kg-1soil applied Cd, this fraction accounted for 15.04, 4.90 and 3.36 percent in treatment involving 5% CaCO3, 40 mg P2O5 kg-1 soil and 2% FYM. This revealed that application of amendments in the form of calcium carbonate (5%) and phosphorus (40 mg P2O5 kg-1) soil helped to increase this form where as FYM decreased this fraction. In limed soils, the activities of free Cd2+ and OH- ions, CO2 partial pressure, control the precipitation of Cd as CdCO 3 (octavite), Cd(OH) and CdCO3 particularly in a sandy soil having low organic matter and low CEC (Street et al 1978). 63

Pierzynksi and Schwab (1993) reported that lime stone treatment significantly increased EDTA-Cd fractions which assumed to represent metal from inorganic precipitates as compared to control. An increase in carbonate bound Cd after lime and P application was also observed by Bolan et al (2003a) and Zwonitzer et al (2003). Singh and Nayyar (1992); Rana and Kansal (1985) also observed that cadmium carbonate controlled the activity of Cd in such soils. After crop harvest CARB-Cd showed very slight decreasing trend. declined marginally with cropping (Tables 4.5.2 and 4.5.3). 4.5.4: Organic bound fraction (OM-Cd) 4.5.4.1 Effect of cadmium: Amount of this fraction at equilibrium increased with increasing rates of Cd steadily from 0.32 in control to 5.24 mg kg-1 soil when it was applied at the rate of 40 mg kg-1 soil at equilibrium. It constituted 8.2, 6.7, 9.7, 10.2, 12.2 and 12.2 percent of the total at 0, 2.5. 5, 10, 20 and 40 mg kg -1 soil in respectively. 4.5.4.2 Effect of Amendments: Amendments had effect of varying magnitude on organic matter bound Cd. Maximum concentration OM-Cd was observed with 2% FYM application and intermediate with 40 mg P kg-1 soil and minimum with 5% CaCO3. At 40 mg Cd kg-1 soil application, the relative percent distribution of Cd in to organic bound fraction was 12.2 percent of the total which decreased to 6.6 (Table 4.4.2) with lime @ 5 % of the total Cd. FYM @ 2% and 40 mg P2O5 P kg-1 soil increased it to 19 and 13.9 % of the total Cd, respectively. Ahumada and Schalscha (1993) observed that Cd was recovered mostly in insoluble organically bound form. The increase due to FYM addition was probably due to addition of Cd (0.38 mg kg-1 FYM) in FYM. Sposito et al (1982) observed that there was an increase in NaOH (organic) Cd with an increase in sludge rates. Pierzynksi and Schwab (1993) studied the sequential fractionation of Cd as influenced by lime stone, cattle manure and poultry manure. Lime stone treatment was clearly the most effective in reducing bioavailable (KNO3, H2O and NaOH) fraction of Cd. But NaOH-Cd was significantly increased by cattle manure and poultry manure as compared to control. With time organic form showed very slight decreasing trend. The corresponding values of the fraction were slightly higher prior to sowing of crop which declined marginally with cropping (Tables 4.5.3). The decline in organic form might be attributed to its conversion into less available fractions. Table No 4.5:3 Distribution of Cd in different fractions as influenced by Cadmium and amendments at post harvest Control The corresponding values of the fraction were slightly higher prior to sowing of crop which

Rates of Cd

EX+WS- CaCO3-Cd Cd

OM-Cd

MnOXCd

A FeOXCd

C FeOXCd

RESCd

64

0 2.5 5 10 20 40

0.04 0.31 0.76 1.74 2.46 3.94

0.10 0.15 0.30 0.54 0.96 1.84

0.10 0.25 0.52 0.94 1.98 4.58

0.68 1.14 1.18 1.58 2.50 6.95

0.98 1.30 1.42 3.22 5.70 9.14

0.58 0.92 1.28 1.38 1.98 4.26

1.36 1.64 2.54 3.98 7.54 12.28

CaCO3 (5 %)

Rates of Cd
0 2.5 5 10 20

EX+WSCd
0.02 0.12 0.28 0.68 0.82 1.46

CaCO3-Cd

OM-Cd

MnOX-Cd

A FeOXCd
1.04 1.38 1.48 3.28 5.98 9.12

C FeOXCd
0.60 0.94 1.38 1.41 1.80 4.34

RESCd
1.25 1.62 2.31 3.94 7.93 12.99

0.23 0.48 0.81 1.89 3.41 6.43

0.06 0.14 0.38 0.64 1.42 2.68

0.74 1.24 1.46 1.64 2.14 6.24

40

FYM (2%)

Rates of Cd
0 2.5 5 10 20 40

EX+WSCd
0.03 0.18 0.42 0.94 1.12 1.96

CaCO3Cd
0.03 0.10 0.15 0.36 0.48 1.38

OM-Cd
0.36 0.52 1.48 2.36 4.88 7.38

MnOXCd
0.78 1.20 1.34 1.80 2.34 7.02

A FeOXCd
1.02 1.36 1.54 3.32 6.12 9.42

C FeOXCd
0.60 0.90 1.28 1.45 1.90 4.62

RESCd
1.04 1.40 1.82 2.58 6.04 10.60

Phosphorus (P2O5 40 mgkg-1 soil)

Rates of Cd
0 2.5 5 10

EX+WSCd
0.03 0.19 0.32 0.78

CaCO3Cd
0.07 0.20 0.36 0.66

OM-Cd
0.16 0.48 0.91 1.34

MnOXCd
0.85 1.15 1.30 1.46

A FeOXCd
0.98 1.27 1.47 3.34

C FeOXCd
0.59 0.94 1.38 1.44

RESCd
1.22 1.74 2.28 4.53

65

20 40

0.98 1.68

1.14 2.10

2.93 5.85

2.34 6.68

5.99 9.42

1.96 4.52

7.87 12.81

66

Fig11: Distribution of Cd in different fractions as influenced by Cadmium and amendments at post harvest 4.5.5: Metals associated with oxides 4.5.5.1Effect of cadmium: At equilibrium, the amount of Cd in manganese oxides (MnOX), amorphous Fe oxides (A FeOX) and crystalline Fe oxides (C FeOX) bound forms increased with increasing rates of cadmium. At 40 mg kg-1 soil application, these forms constituted about 13.4, 20.3 and 9.7 per cent respectively of the total Cd in soil. Relative higher value of oxides (MnOX, A FeOX and C FeOX) fractions compared to the others fractions suggested that Cd was mostly found occluded in these fractions. Keller and Vedy (1994) indicated that Cd was mainly associated with Fe- Mn oxides. Bruemmer et al (1988) suggested that higher amount of Cd in this fraction was due to its specific adsorption on Fe-Mn oxides or incorporated inside the oxide particles. Jenne (1968) reported that concentration of heavy metals was inclined to be controlled by sorption to hydrous oxides After harvest, with time, amount of Cd bound to MnOX, A FeOX and C FeOX ) were 6.95, 9.14 and 4.49 mg kg -1soil 67

which were higher compared to their values of 5.79, 8.74, 4.18 mg kg-1soil of the above fractions respectively form. 4.5.5.2 Effect of Amendments: At equilibrium, increasing trend for all oxide (MnOX, A FeOX, C FeOX) fractions was observed with all the added amendments. However a slight difference in Fe, Mn bound Cd existed among amendments. Amount of MnOX-Cd increased from 13.42 to 14.08 percent with the application of calcium carbonate. Same trend was found in A FeOX-Cd which increases from 20.26 to 20.75 and in C FeOX fraction from 9.69 to 9.84 percent. Application of FYM encouraged Cd to be accumulated more in this fraction FYM @ 2 % registered higher values of all oxide bound fractions (6.68, 9.32 and 4.48 mg kg-1soil ) as compare the equilibrium (5.79, 8.74, 4.18 mg kg
-1

at equilibrium. Logan and Chaney (1983) indicated that over time a

Cd salt solution reacted with soil and reverted to less available or more insoluble oxidised

soil). This was most likely due to

predominance of dithionite extractable iron and manganese oxides (Free Fe and Mn oxide ) having a mean value of 0.64 per cent and 156 mg kg-1 soil. Shuman (1988) reported that amorphous Fe oxide Zn increased with the application of organic matter. Application of phosphorus @ 40 mg kg-1soil resulted in the increase of Cd bound pools MnOX-Cd, A FeOXCd and C FeOX-Cd to 6.62, 9.18 and 4.44 mg kg-1soil respectively from 5.79, 8.74, 4.18 mg kg-1soil. After harvest of crop, amount of Cd bound to MnOX, A FeOX and C FeOX ) were 6.68, 9.42 and 4.52 mg kg -1soil as against 6.62, 9.18 and 4.44 mg kg-1soil indicating thereby that cropping did no affect this form. Ghafoor et al (2008) also reported increase in FE/Mn bound fractions with KH2PO4 and lime.

4.5.6: Residual fraction (RES-Cd) 4.5.6.1 Effect of cadmium: There was an increase in residual Cd with increasing level of Cadmium. Application of 40 mg kg-1 soil increased the amount of Cd in this pool from 1.39 to12.60 mg kg-1. 4.5.6.2 Effect of amendments: Lime application influenced this fraction and encouraged Cd to be more associated with this fraction. However P application ineffectually increased residual Cd in soil. Application of FYM inefficiently decrease this fraction. This is in conformity with Chen et al (2000), Bolan et al (2003a & b) and Zhu et al (2004) who reported that lime, P and compost increased residual Cd in soil. It is assumed that metal fraction lost from the Exch+WS was recovered in the /oxides and residual fractions (Knox et al 2001) The results of the present study in the preceding pages indicated that among the three amendments (FYM, Calcium carbonate and Phosphorus), calcium carbonate was found to be 68

most effective in mitigating the toxicity of cadmium followed by phosphorus and FYM. It is quite possible that application of calcium carbonate may cause deficiency of some essential micronutrients which is a cause of concern. Considering soil health as of paramount and vital importance, application of FYM appears to most suitable amendments as it not only offset the adverse effect of cadmium but also causes improvement in soil structure by increasing porosity, water holding capacity and aeration. Other useful constituents of FYM also help to sustain soil productivity. Further, there is need to characterize organic matter as in some cases higher fulvic content encourage plant uptake of pollutants. Application of phosphorus may also antagonize zinc availability under certain situations. Considering the above facts, choice of amendments has to be made carefully. So, there is need to conduct further experiments to study the

effect different types of

both inorganic and organic ameliorants under specific

situations depending on the severity of pollutant toxicity so that their availability could be kept at the minimum.

CHAPTER V SUMMARY There is unabated contamination of soils with heavy metals including Cd through sewage water, city garbage, and untreated industrial effluents as a result of rapid industrialisation and urbanisation in Punjab in the recent past. High amount of cadmium has been reported to cause health hazards and disorders in human beings. In Punjab, leafy vegetables including Chalai (Pig weed- Amaranthus tricolor) are generally grown on the outskirt of the cities and irrigated frequently by sewage water contaminated largely by industrial effluents. There is need to develop appropriate decontamination technology using range of inorganic and organic materials for minimizing its pollution in agricultural soils and crops grown on these soils. There was paucity of information on the transformation and availability of Cd in the presence and absence of amendments (lime, phosphate and organic manure). In view of all this, objective of the of the present study were; 69

1. To delineate Cd contaminated soil for its lateral distribution in order to assess its
extenent pollution. 2. To assess relative suitability of different amendments for minimizing Cd pollution in contaminated soils and plants. 3. To establish the upper threshold limit of toxicity of Cd for pig weed. 4. To study the effect of various amendments on the transformations of Cd in soil. Survey Studies In order to determine pollution potential, surface (0-15cm) soil samples receiving waste water irrigation at a distance of 50, 250, 500, 750 and 1000 meters (5 sites) were collected laterally on either side (Right and left) along the Buddah Nallah from each village (six in number) which were approximately 4-5 kilometers from each other with the help of global positioning system in the month of December. These were analysed for DTPA and total Cd Mean value of DTPA-Cd irrespective of sites of surface sewage irrigated soils was 0.154 mg kg-1 soils which was found to be 5.2 times greater than the normal soils In order to demarcate Cd polluted soils, guidelines based on total metal content were considered. As per guidelines of Kabata and Pendias (1984), 3-8 mg total Cd kg -1 soil is considered to be the critical limit above which toxicity of Cd is possible. In the present study, 6 (representing the sites Saleem tabri and Partapsinghwala) out of 53 soil samples reached this threshold value of 3 mg Cd/kg soil. Thus it appeared that about 11.3 percent soils have become polluted as a result of continuous irrigation with sewage water. These soils require immediate attention and needs to be ameliorated urgently. There is the possibility that rest of the soils might approach this critical limit in a few years if same level of irrigation with sewage water continued. In another standard adopted in U.K, as prescribed by G L.C (Greater London Council), total Cd concentration ranging from 0 to 1, 1 to 3, 3 to 10, 10 to 50 and >50 mg/kg soil were categorized as typical uncontaminated, slightly contaminated, contaminated and heavy contaminated soils, respectively. According to this system therefore, most of the investigated soils fell under slightly contaminated category indicating thereby that clean up operation is definitely required. So there is an urgent need to work out the critical limit for Cd toxicity in the Punjab soils on a wider scale Screen house studies A screen house experiment was conducted to assess the effect of six levels of Cd (0, 2.5, 5 10, 20 and 40 mg kg-1 soil) and CaCO3 (2.5 and 5%), FYM (1 and 2 %) and Phosphorous (20 and 40 mg kg-1 soil) in a factorial complete randomized design on the growth of pigweed on sandy loam soil having DTPA extractable Cd 0.36 mg kg-1 soil. The

70

soil in the pots was equilibrated for 21 days at field capacity moisture level after applying above treatments. The crop was then planted and harvested at grand growth stage. Dry matter yield of shoot and roots the crop was recorded and plant samples of both shoots and roots were analyzed for Cd and micronutrients cations. The soil samples collected after equilibrium and crop harvest were analyzed for DTPA- extractable Cd. A laboratory study was also conducted to determine different pools of cadmium and their transformations as influenced by amendments. The result of the screen house study revealed that with the increasing rates of Cd application, there was significant and progressive increase in DTPA -Cd at equilibrium. The application CaCO3, FYM and phosphorous declined the content of DTPA- Cd in soils at all level of its application. DTPA-Cd decreased from 7.38 mg kg-1 soil ( control) to 3.8 (lime @ 2.5%), 3.5 (lime @ 5%); 4.9 ( FYM @ 1% ), 4.64 (FYM @ 2% ); 4.4 mg kg-1 soil (P @ 20 mg P kg-1 soil) and 4.0 mg kg-1 soil (P @ 40 mg P kg-1 soil), thereby indicating the superiority of applying CaCO3 as an ameliorant over other amendments for mitigating the toxicity of cadmium Cadmium content in the shoots increased with increasing rates of Cd application. The application of even the lowest rate of 2.5 mg Cd kg-1 soil significantly increased the Cd content in crop over control. The mean Cd concentration increased from 1.26 in control treatment to 4.72, 6.04, 14.10, 21.80 and 46.42 g g-1 dry matter when rate of Cd application was raised to 2.5, 5, 10 20 and 40 mg kg-1 soil. It implied now that the decrease in dry matter yield had resulted from the phyto toxic effect of Cd in pigweed emanating from the increased availability of Cd in soil and plants as a result of its application. A gradual reduction in mean dry matter yield of chalai (pig weed) occurred with increasing levels of cadmium irrespective of the amendments but the significant decrease was observed at and above the application rate of 10 mg kg-1soil. However different amendments viz (Calcium carbonate, FYM, Phosphorus) exhibited variable behaviour as far as their remediation potential was concerned at a particular level of Cd application and their application at all the levels mitigated the toxicity of cadmium as is evident by the increase in dry matter yield of the crop irrespective of Cd levels. A significant enhancement in dry matter yield of chalai was observed with all the amendments. Lime @ 2.5 and 5% increased mean dry matter yield from 16.39 (Control) to 18.2 and 19.1 g pot-1, respectively. FYM at the rate 1 and 2% produced mean dry matter yield of 18.4 and 19.34 g pot-1 while 17.6 and 18.4 g pot-1 respectively with phosphorus application of 20 and 40mg kg-1 soil regardless of Cd levels. The dry matter yield of roots started declining even with the application rate of 5 mg Cd kg-1 soil application. However, a significant decrease was observed with 10 mg Cd kg-1 soil onwards. Further, application of 20 and 40 mg Cd kg-1 soil showed significant sharp decline in root dry matter yield. The 71

reduction in dry matter yield of pigweeds root was 15.9, 28.2 and 45.7 percent with the application of cadmium at the rate of 10, 20 and 40 mg kg-1soil. So the yield reduction in response to Cd was of similar proportions in the roots as for the aboveground part of the plants. The reduction in root growth was in accord with the amounts of extractable Cd .The magnitude of reduction in root biomass was comparable to the shoot biomass. Maximum mean dry matter yield of the shoots of pig weed was obtained in the treatments involving FYM amendments in spite of the fact it was less effective in decreasing the concentration of Cd in pig weed shoot compared to other amendments. Amendment in the form of calcium carbonate was found to be most effective. With amendments (calcium carbonate) through formation of less soluble compounds like CdCO3 mitigated the toxicity of cadmium. There was excellent fit to the data with quadratic model as revealed by significant coefficient of determination (R2) From these equations, the toxic level of DTPA Cd at which 20 percent reduction in dry matter yield occurred, were then estimated. The toxic levels of DTPA Cd was found to be 4.38 mg kg-1 soil for pig weed The toxic level of Cd in shoots of pig weed at grand growth stage through quadratic model was found to be 14.6 g g-1 dry matter. Applied cadmium at lower levels (0 to 10mg Cd kg-1 soil) increased Zn concentration in roots and shoots of crop showing synergistic interaction. It indicated that, one helps in the absorption of the other at lower levels. However at higher levels (20 and 40 mg kg-1 soil) of Cd application, Zn concentration decreased relative to control indicting antagonistic interaction. Like zinc, mean Fe concentration in the roots and shoots of crop showed increasing trend with the application of Cd at lower levels but depression at higher levels. The concentration of copper and manganese in the roots and shoots of pig weed was negatively affected by the application of Cd at all levels as evident from the significant negative correlation coefficients between these metals and cadmium. Different pools of Cadmium as influenced by Cd and amendments application at equilibrium All the fractions of Cd increased significantly with increasing levels of cadmium irrespective of amendments at equilibrium before seeding of crop. Application of amendments had a depressing effect on the amount of cadmium in the EX+WS fraction. Manure application decreased the amount of cadmium in carbonate where as it encouraged the Mn oxide, organic bound and amorphous Fe oxide crystalline FeOX bound Cd. Application of calcium carbonate at the rate 5% increased the amount of carbonate substantially but had a non significant effect on all other forms. Phosphorus application was effective in trasnsforming Cd in to less available forms (Mn oxide, amorphous Fe oxide and crystalline FeOX bound Cd). Different pools of Cadmium as influenced by Cd and amendments application at post harvest 72

There was a decreasing trend of extractability of exchangeable + water soluble, carbonate bound and organic bound fraction with time both in the presence and absence of amendments. The decline in these forms might had been caused partly by its progressive removal by plants or due to possible shift to other forms. In all the treatments (with and without amendments) with time/cropping there was an increasing trend of Cd to be associated more in the MnOX, A FeOX and C FeOX fractions. The value of this fraction was comparatively higher after the harvest of the crops compared to at equilibrium.

REFERENCE
Abdel-Sabour M F, Mortvedt J J and Kelso J J (1988) Cadmium-zinc interactions in plants and extractable cadmium and zinc fractions in soil. Soil Sci 145: 424-31. Adhikari T, Tel E. Libal Y and Shenker M (2006) Effect of cadmium and iron on rice (Oryza sativa L.) plant in chelator-buffered nutrient solution. J Pl Nutr 29: 191940. Aghabarati A, Hosseini S M and Maralian H (2008) Heavy metal concentration of soil and olive trees (Oleaeuropaea L.) In Suburban areas of Tehran, Iran. Res J Environ Sci 2: 323-29. Ahumada I T and Schalscha E B (1993) Fractionation of cadmium and copper in soils. Effect of redox potential. Agro chemical 37: 281-89. Alekseev A A and Zyrin N G (1982) Behaviour of cadmium in soil plant system. Soil Sci 37: 21-28.

73

Alloway B J, Jackson A P and Morgan H (1990) The accumulation of cadmium by vegetables grown on soils contaminated from variety of sources. Sci Total Environ 91: 233-35. Arauja D O, Nascimentto C W and Pareira J E M (1997) Uptake and distribution of cadmium and micronutrients by bean cultivars exposed to cadmium levels. Pesquisa Agropecuaria Brasileira 32(12): 1303-08 Artexe U, Gareias-Plazoals J L, Hernandez A and Becerril J M (2002) Low light grown duckweed plants are more protected against the toxicity induced by Zn and Cd. Pl Physiol Biochem 40: 859-63. Asami T, Kubata M and Orikasa (1995) Distribution of different fraction of cadmium, zinc, lead and copper in unpolluted and polluted soils. Water, Air and Soil pollut 83: 18794. (Original not seen, Abstr in CAB Abstract A.N 951911659). Atiyeh R M, Edward C A, Subler S and Metzger J D (2001) Pig manure, vermicompost as a component of horticulture bedding plant medium: effect on physiochemical properties and plant growth. Bioresource Technol 78: 11- 20. Basta N T and McGown S L (2004) Evaluation of chemical immobilization treatments for reducing heavy metal transport in a smelter-contaminated soils. Environ Pollut 127: 383-86. Basta N T, Gradwohl R, Snethen K L, Schroder J L (2001) Chemical immobilization of lead, zinc and cadmium in shelter contam, inated soils using biosolids and rock phosphate. J Environ Qual 30: 1222- 30. Bell P, Bruce R J and Chaney R L (1991) Heavy metal extractability in long term sewage sludge and metal salt amended soils. J Environ Qual 20: 481-86. Berthet B, Amiard J C, Amiard-Triquet C and Metayer C (1984) A study of biogeochemical cycle of cadmium using experimental plant culture system and application of the results to the agriculture use of sewage sludge from treatment plants. Environ Pollut 8: 35-50. Bipasha C, Srivastava S, Chakravarty V and Srivastava B (1997) Effect of cadmium and zinc interaction on metal uptake and regeneration of tolerant plants in linseed. Agric Eco Environ 61(1): 45-50. Bolan N S, Adriano D C, Duraisamy P, Mani P A and Arulmozhiselvan K (2003a) Immobilization and phytoavailability of cadmium in variable charge soils. I. Effect of phosphate addition. Pl Soil 250: 8394. Bolan N S, Naidu R, Khan M A R Tillman and Syers J K (1999) The effect of anion sorption on sorption and leaching of cadmium. Aust J Soil Res 37: 445-60. Bolan, N S, Adriano D C, Mani P A and Duraisamy A (2003b) Immobilization and phytoavailability of cadmium in variable charge soils. II. Effect of lime addition. Pl Soil 251: 18798. Bolland M D, A,Posner A M and Quirk J P (1977) Zink adsorption by goethite in absence and presence of phosphate. Aust J Soil Res 15: 279-86.

74

Borges A C and Wollum A G (1981) Effect of cadmium on symbiotic soyabean plants. J Environ Qual 10: 216-21. Brar M S, Khurana M P S and Kansal B D (2002) Effect of irrigation by untreated sewage effluents on the micro and potential toxic elements in soils and plants. Proc 17th World Congress Soil Sci, Bangkok, Thailand 4: 198(1-10). Brown S L, Chaney R, Hallfrisch J, Ryan J A and Berti W R (2004) In situ soil treatments to reduce the phyto and bioavailability of lead, zinc and cadmium. J Environ Qual 33: 522-31. Brown S, Chaney Q C, Angle J S and Ryan J A (1998) The phytoavailbility of cadmium to lettuce in a long term bio solid amended soils J Environ Qual 27: 1071-78. Bruemmer G W, Gerth J and Tiller K G (1988) Reaction kinetics of adsorption and desorption pf nickel, zinc and cadmium by goethite. Adsorption and diffusion of metals. J Soil Sci 39: 37-52 Cao R X, Ma L Q, Chen M, Sati P S and Harris W S (2003) Phosphate-induced metal immobilization in a contaminated site. Environ Pollut 22: 19-28. Cataldo D A, Garland T R and Wildung R E (1981) Cd distribution and chemical fate in soybean plants. Pl Physiol 68: 835-39. Cataldo D A, Garland T R, Wildung R E (1983) Influence of soil applied Cd on growth and nutrient composition of plant species. Pl Physio 73: 84448. Chao T T (1972) Selective dissolution of manganese oxides from soils and sediments with acidified hydroxylamine hydrochloride. Soil Sci Soc Am Proc 36: 764-68. Chao T T and Zhou L (1983) Extraction techniques for selective dissolution of amorphous iron oxides from soil and sediments. Soil Sci Soc Am J 47: 225-32. Chen H M, Zheng C R, Tu C and Shen Z G (2000) Chemical methods and phytoremediation of soil contaminated with heavy metals. Chemosphere 41: 22934. Chen M, Ma L Q, Singh S P, Cao R X and Melamed R (2003) Field demonstration of in situ immobilization of soil Pb using P amendments. Adv Environ Res 8: 93-102. Chen X, Wright J V, Conca J L, Perurrung L M (1997) Evaluation of heavy metal remediation using mineral apatite. Water Air Soil Pollut 98: 57 -78. Chen Z, Zhao Y, Li Q Jiejuan Q, Tian Q and Liu X (2009) Heavy metal contents and chemical speciations in sewage-irrigated soils from the eastern suburb of Beijing, China. J Food Agric Environ 7: 690-95. Chernykh N A (1991) Alteration of the concentration of certain elements in plants by heavy metals in the soil. J Ind Soc Soil Sci 23(6): 45-53. Christensenn, T H (1984) Adsorption of Cd as influenced various soil properties. Water Air Soil Pollut 21: 10510.

75

Cieslinski G, Neilson G H and Hogue E J (1996) Effect of soil cadmium application and pH on growth and cadmium accumulation in root, leaves and fruit of strawberry plants. Pl Soil 180: 267-76. Clemente R Peredes C and Bernal M P (2007) A field experiment investigating the effects of olive husk and cow manure on heavy metal availability in contaminated calcareous soils from Murcia (Spain) Agri Ecosystem Environ 118: 319-26. Cottenie A, Camerlynck R, Verloo M and Dhaese A (1979) Fractionation and determination of trace elements in plant, soils and sediments. Pure Applied chem 52: 45-53. Dahiya S S, Geol S, Antil R S and Karwasra S P S (1987) Effect of farm yard manure and cadmium on the dry matter yield and nutrient uptake by maize. J Ind Soc Soil Sci 35: 460-64. Datta S P, Rattan R K and Aiando S (2007) Influence of different amendments on theavailability of cadmium to crops in the sewage irrigated soils. J Ind Soc Soil Sci 55: 86-89. Dheri G S Brar M S and Mahi S S ( 2007) Influence of phosphorus application on cadmium uptake of spinach in twotwo cadmium contaminated soils. J Pl Nutr Soil Sci 170: 1-5. Dudka S and Chlopecka A (1991) Mobility and phyto availability of cadmium and zinc as related to their species on their sludge amended soils. 6thInt Trace Element Symp pp 1395-1402. (Original not seen, Soil and Ferti. abstract 54 Abstract No. 1166) Ebbs D and Kochian V (1998) Toxicity of zinc and copper to Brassica species: Implications for phytoremediation. J Environ Qual 26(3): 776-81. El-Arby A M and Elbordiny M M (2006) Impact of reused wastewater for Irrigation on availability of heavy metals in sandy soils and their uptake by plants. J Applied Sci Res 2: 106-11. Emmerich W E, Lund L J, Page A L and Chang A C (1982) Solid phase form of heavy metals in sewage sludge treated soils. J Environ Qual 11: 178-81. Friesel W, Horak O and Wenzel W W (2004) Immobilization of heavy metals in soils by the application pf bauxite residues: Pot experiments under field conditions. J Pl Nutr Soil Sci 167: 16. Ghafoor A, Muhammad Z R R, Murtaza G and Sabir M ( 2008) Fractionation and Availability of Cadmium to Wheat as Affected by Inorganic Amendments. In J Agri Biol 10: 46974. Gibson M J and Farmer J G (1986) Multi step sequential chemical extraction of heavy metal from urban soils. Environ Pollut 11: 117-135. Greger M (1999) Salix as phytoextractor. Proc 5thIntConfBiogeochem Trace Elements, Vienna, 99. Griffiths J R and Wardsworth (1980) Heavy metal pollution on farms near an industrial complex. In: Inorganic Pollution and Agriculture. pp 70-76. London. 76

Guo G, Zhou Qand M A L Q (2006) Availability and assessment of fixing additives for the in-situ remediation of heavy metal contaminated soils. Environ Monitoring assessment 116: 51328. Gupta A P, Narwal R P and Singh J P (1989) Effect of farm yard manure on the yield and heavy metal accumulation in wheat grown in a polluted soil.In Proc National Symposium on Impact and Management on Crop Productivity. pp 86-87. Haryana Agricultural University. Hissar, India. Gupta S K and Chen K Y (1975) Partitioning of trace metals in selective chemical fractions of near shore sediments. Environ Lett 10: 129-58. Gupta S K and Mitra A (2002) In: Advances in Land Resource Management for 21stCentury, Soil Conservation Society of India, New Delhi, p.446-460. Gupta V K and Gupta S P (1988) Effect of gypsum and zinc sources on yield and Zn, Cu, Mn and Fe nutrition of soybean (Glycine max L.) on sodic soils. Ann Arid Zone 24: 16276. Gupta V K and Dixit M L (1992) Influence of soil applied cadmium on growth and nutrient composition of plant species. J Ind Soc Soil Sci 40: 87880. Gupta S, Satpati S,Nayek S and Garai D (2007) Effect of wastewater irrigation on vegetables in relation to bioaccumulation of heavy metals and biochemical changes. Environ monitoring and Assessment 165(1-4): 169-77. Haghiri F (1973) Cadmium uptake by plants. J Environ Qual 2: 93-96. Han D H and Lee J H (1996) Effects of liming on uptake of lead and cadmium by Raphanus sativa. Archives Environ Contami Toxicol 31: 488-93. Haq A U, Bates T E and Soon Y K (1980) Comparison of extractants for plant available zinc, cadmium, nickel and copper in contaminated soil. Soil Sci Soc Am J 44: 772-77. Hettiarachchi G M and Pierzynski G M (2002) In situ stabilization of soil lead phosphorus and manganese oxide: Influence of plant growth. J Environ Qual 31: 564-72. Hiroyuki T, Wataru O, Eiji N, and Masaharu M ( 2010) Reduction of cadmium uptake in spinach (SpinaciaoleraceaL.) by soil amendment with animal waste compost. J Hazard Materi 181: 298-04 Iwai I, Hara T and Sonoda Y (1975) Factors affecting cadmium uptake by the corn plant. Soil Sci Pl Nutr 21: 37-46. Iwegbue C M A, Emuh F N, Isirimah N O and Egun A C (2007) Fractionation, characterization and speciation of heavy metals in composts and compost-amended soils. Afr J Biotechnol 6 (2): 67-78. Jackson ML (1973) Soil chemical analysis. Prentice Hall of India Pvt Ltd, New Delhi. Jasiewicz C (1993) Pollution of vegetables with heavy metals. Zeszyty Neukowe Akademii Rolniczej in Hugona Kollataja W Krakowie. Rolnictwo 30: 129-43. 77

Jeng A S and Bergseth H (1992) Chemical and mineralogical properties of Norweign alum shale soils with special emphasis on heavy metal content and availability. Acta Agric Scand Sec 42: 88-93. Jeng A S and Singh B R (1993) Partitioning and distribution of cadmium and zinc in selected cultivated soils in Norway. Soil Sci 4: 240-50. Jenne E A (1968) Control on Mn Fe, Co, Ni and Zn concentration in soil and water. The significance role of hydrous oxides. In R A Baker (ed) Trace Inorganic in Water. Adv Chem Ser 73: 337-87. Jing, J, and Logan T J (1992) Effects of sewage sludge cadmium in agricultural soils. J Enviorn Qual 26: 96674. John M K VanLaerhoven C J and Chukwuma C S (1972) Factors Affecting Plant Uptake and Phytotoxicity of Cadmium Added to Soils. Environ Sci and Techno l6(12):1005-09 Juwarkar A S and Shende G B (1986) Effect of cadmium enriched sewage sludge on the growth and cadmium content of wheat in a verti soil. J Ind Soc Soil Sci 34: 539-47. Kabata P A and Pendias H (1984) Trace Elements in Soil and Plants. p365. CRC Press Inc Boca Raton, Florida, U.S.A. Kabata Pendias A. In Heavy metals ( Problems and Solutions) Ed . W Salomons, U Forstner and P Mader) Spring veriag. Berlin, Heidelberg, New York, London , Tokyo, 3-18 (1995) Kansal B D and Khurana M P S (1999) Extent of contamination of alluvial soils with cadmium.In. 2nd International conference on Contaminants in soil environment in Australia Pacific region, New Delhi.pp:309-10. Keller C and Vedy J C (1994) Distribution of copper and cadmium fractions into two forest soils. J Environ Qual 23: 987-99. Khan D H and Frankland B (1983) Chemical form of cadmium and lead in some contaminated soils. Environ Pollut 6: 15-31. Khan D H and Frankland B (1983) Effects of cadmium and lead in the radish plants with particular reference to movement of metals through soil profile and plant. Pl Soil 70: 335-45. Khan S and Khan N N (1983) Influence of lead and cadmium on the growth and nutrient concentration of tomato (Lycopersicon escolentum) and egg plant (Solanum melongena). Pl Soil 74: 387394. Khurana M P S and Kansal B D (2000) Bio-availability of cadmium to maize crop as influenced by cadmium and farm yard manure. In Proc 65th Annual Convention of Ind Soc Soil Sci, pp 112.Nagpur, India. Khurana M P S, Nayyar, V K and Bansal R L (2006) Relative performance of maize cultivars for their tolerance to cadmium. Environ & Ecology 24: 270-74.

78

Khurana MPS, Nayyar VK and Bansal RL (2003) Crop plants for phytoremediation of agricultural lands contaminated with heavy metals. Proc National Confon Soil-Elixir of Life.Patiala Pp 38. Knox A S, Seaman J C, Mench M J and Vangronsveld J (2001)Remediation of metalandradionuclides- contaminated soils by insitustabilization techniques. In: Iskandar, I.K. (ed.), Environmental Restoration of Metals-Contaminated Soils, pp:2160. CRC Press, Inc.,Boca Raton, FL, USA Koreak R F and Fanning D S (1978) Extractibility of cadmium, copper, nickel and zinc by double acid vs DTPA and plant content at excessive soil level. J Environ Qual 7: 506-12. Koshino M (1973) Cadmium uptake by rice and wheat as affected by the application of phosphate and some metal elements. Bulletin Nation Inst Agr Sci, Tokyo, Japan, pp. 24151. Kuhad M S, Malik R S and Singh A (1989) Background levels of heavy metalsin agricultural soils of IndoGangetic Plains of Haryana. J Ind Soc Soil Sci 37: 70005. Kumpiene J, Lagerkvist A, Maurice C (2007) Stabilization of Pb- and Cu contaminated soil using coal fly ash and peat Environ Pollut 145: 365-73. Kuo S, Heilman P E and Baker A S (1983) Distribution and forms of copper, zinc, cadmium, iron and manganese in soils near a copper smelter. Soil Sci 135: 101-09. Krsad Trkdo M, Kilicel F, Kara K, Tuncer I and Uygan I (2003) Heavy metals in soil, vegetables and fruits in the endemic upper gastrointestinal cancer region of Turkey. Environ Toxicol and Pharmacol 13 (3): 175-179. Lake D L, Kirk P W and Lester I N (1984) Fractionation, characterization, speciation of heavy metals in sewage sludge and sludge amended soils. J Environ Qual 13: 175183. Lakhdar A, Achiba W, Montemurro F, Jedidi N, and Abdelly C (2008) Effect of Municipal Solid Waste Compost and Farmyard Manure Application on Heavy-Metal Uptake in Wheat. Commun Soil Sci Pl Anal 40: 3524-38 Lamoreaux R J and Chaney W R (1977) Growth and water movement in silver maple seedling affected by cadmium. J Environ Qual 6: 201-5. Laperche V, Traina S J, Gaddam P and Loggon T J (1996) Chemical and mineralogical characterizationof Pd from contaminated soils:Reactions with synthetic apatite. Environ Sci Technol 30: 3321-26. Larlson, J, Likens G, Fitzpatrick J and Crock J (2000) Cadmium Toxicity Among Wildlife in the Colorado Rocky Mountains. Natur 406: 181-83. Lee K C, Cunninghum B A, Paulson G M, Liang G N and Moor R B (1976) Effect of cadmium on respiration rate and activity of several enzymes in soybean seedlings. Pl Physiol 36: 4-6.

79

Lee T M, Lai HY and Chen Z S (2004) Effect of chemical amendments on the concentration of cadmium and lead in long-term contaminated soils. Chemosphere 57: 145971. Lehoczay E, Szabados I and Marth P (1996) Cadmium content of plants as affected by soil cadmium concentration. Commun Soil Sci Pl Anal 27(5-8): 1765-77. Lehoczky E, Szabo I, Horvath S, Martin P and Szabo I (1998) Cadmium uptake of lettuce in different soils. Commun Soil Sci Pl Anal 29: 11-24. Levi-Minzi R and Petruzzelli G (1984) The influence of phosphate fertilizers on Cd solubility in soils. Water Air Soil Poll 23: 423-29. Lindsay WL and Norvell WA (1978) Developmental of a DTPA soil test for Zn, Fe, Mn and Cu. Soil Sci Am J 42: 421-28. Liu W, Zhao J, Ouyang Z, Sderlund L and Liu G (2005) Impacts of sewage irrigation on heavy metal distribution and contamination in Beijing, China. Liu Y H, Lee F Y and Liao C J (1997) The sequential fractional and desorption of copper and cadmium in several Taiwan soils. J Chinese Agric Chem Soc 35 : 683-91 Original not seen, Abstr in CAB Abstract A.N 981907251). Logan T J and Chaney R (1983). Metals. In A. L. Page (ed.) Utilization of Municipal Wasteland and Sludge on Land. pp 235-329. University of Califorina, Riverside, USA. Lokeshwari H and Chandrappa G T (2006) Impact of heavy metal contamination of Bellandur Lake on soil and cultivated Vegetation. Curr Sci 91: 622-27. Ma L Q and Gade N R (1997) Effects of phosphate rock on sequential chemical extraction of lead in contaminated soils. J Environ Qual 26: 788-94 Ma Y B, and Uren N C (1998) Transformation of heavy metals added to soil- Application of new sequential extraction procedure. Commun Soil Sci Pl Anal 19: 1747-70. Maclean A J (1976) Cadmium in different plant species and its availability in soils as influenced by organic matter and addition of lime, phosphorus, cadmium and zinc. Can J Soil Sci 56: 129-38. Mahler R J, Bingham F T, Page A L (1978) Cadmium enriched sewage sludge application to acid calcareous soils. Effect on yield and cadmium uptake by lettuce and chard. J Environ Qual 7: 274-81. Mahler R J, Bingham F T, Page A L and Ryan J A (1982) Cadmium enriched sewage sludge application to acid calcareous soils. Effect on soil and nutrition of lettuce, corn, tomato and swiss chard. J Environ Qual 11: 694-700. Mapanda F, Mangwayana E N, Nyamangara J and Giller K E (2005) The effect of long term \irrigation using wastewater on heavy metal contents of soils under vegetablesin Harare, Zimbabwe. Agric Ecosyst Environ 107:151-65.

80

Mark E, and Eugenia J (2000) Can metal phosphate formation in soils be used as a treatment for metal contaminated soil? Land Contamination & Reclamation, 8 (3), DOI 10.2462/09670513.563 Matusik J, Bajda T and Manecki M (2008) Immobilization of aqueous cadmium by addition of phosphates. J Hazard Materi 152: 1332-39. McBride M B (1995) Toxic metal accumulation from agriculture use of sludge. Are USEPA regulations productive. J Environ Qual 24: 5-18. McGrath S P, Zhao F J and Lombi (2002) Phytoremediation of metal, metalloids and radionuclides. Adv in Agron 75: 1-56. McLaren R G and Crawford D V (1973) Studies on soil copper. The fractionation of copper in soils. J Soil Sci 24: 172-81. Mehra O P and Jackson M L (1960) Iron oxide removal from soils and clays by a dithionitecitrate system buffered with sodium bicarbonate. Clay Miner 7: 317-27. Miller W and McFee W W (1983) Distribution of cadmium, zinc, copper and lead in soils of industrial north western Indiana. J Environ Qual 12: 29-33. Mishra A, Tripathi B D (2008) Heavy metal contamination of soil and bioaccumulation invegetables irrigated with treated waste water in the tropical city of Varanasi, India. Toxicol Environ Chem 90: 86171. Mitra A and Gupta S K (1999) Effect of sewage water irrigation on essential plant nutrient and pollutant element status in a vegetable growing area around Calcutta. J Ind Soc Soil Sci 47: 99-104. Mohammad M J, Mazahreh N (2003) Changes in soil fertility parameters in response to irrigation of forage crops with secondary treated wastewater. Commun Soil Sci Pl Anal 34: 1281-94. Naidu R, Kookuna R S, Oliver D P, Rogers S and McLaughlin M J (1996) Contaminants and the soil environment in the Austrilasia-Pacific Region. Proceedings of the First Australasia-622 RANI FARYAL ET AL.,Pacific Conference on Contaminants and Soil Environment in the Australasia-Pacific Region, held in Adelaide, Australia 20-24 February 1996. Kluwer Acedamic Publishers. 629-46. Narwal R P and Singh B R (1998) Effect of organic materials on partitioning, extractability and plant uptake of metals in an alum shale soil. Water Air and Soil Pollut 103: 40521. Narwal R P, Singh M, Singh J P, Dahiya D J (1993) Cadmiumzinc interaction in maize grown on sewer water irrigated soil. Arid Soil Res Rehab 7: 125-35. Narwal RP, Singh M and Dahiya DJ (1990) Effect of cadmium on growth and heavy metal content of corn ( Zea mays L ) Crop Res 3: 13-20. Obata H, Inouhe N and Umebayashi M (1996)Effect of Cd on plasma membrane ATOase from plant and roots differing in tolerance to Cd. Soil Sci Pl Nutr 42: 361-66. 81

Ochiai E I (1995) Toxicity of heavy metals and biological defence. J Chemi Edu 72: 47984. Okamoto T, Hirobe M, Wachi K and Matsuzaki T (1990) Changes in form mobility and availability of some heavy metal in a soil with long term application of sewage sludge. Proc Int Congr Soil Sci 4: 216-21. Oliver D P, Hannam R, Tiller K G, Wilham N S, Mrrry R H and Cozen G D (1994) The effects of zinc fertilization on cadmium concentration in wheat grain. J Environ Qual 23: 705-711. Page AL (1981) Cd in soils and its accumulation by food crops. In: Heavy metals inthe environment, Edinburgh, U.K. CEP Consultants Ltd 206-13. Panse VG and Sukhatme PV (1967) Statistical methods for Agricultural Workers.Model (Pvt) Press, New Delhi. Panwar B S, Dahiya D S, Singh R, and Kuhad M S (2001) Phyto availability of Cd and P in brassica species grown in soil amended with Cd and P fertilizers. Sustain Use Chemi Agric 2: 12-17. Panwar B S, Singh J P and Laura R D (1999) Cadmium uptake by cowpea and mung-bean as affected by Cd and P application. Water Air Soil Pollut 112: 163-69. Patel K P, Pandya R P, Maliwal G L, Patel K C, Ramani V P and George V (2004) Heavy metal content of different effluents and their relative availability in soil irrigated with effluent water around major industrial cities of Gujarat. J Ind Soc Soil Sci 52: 89-94. Patel P M, Wallace A, Hortsock T and Romney E M (1980) Zn, Ni, Cd uptake and translocation to seed pods and their effects on gas exchange rates of bush bean plants grown in calcareous soil from the Northern Mojave Desert. J Pl Nutr 2: 67-72. Patel P M, Wallace A, Mueller R T (1976) Some effects of Cu, Co, Cd, Zn, Ni and Cr on growth and mineral element concentration in Chrysanthemum. J Am Soc Hort Sci 101: 5356. Peles J, BrewerS, and Barrett G (1998) Heavy Metal Accumulation by Old-field Plant Species During Recovery of Sludge-treated Ecosystems. Am Midland Natur 140(2): 245- 51. Perez-Murcia M D Moral R Moreno-Caselles J, Perez-Espinosa and Paredes C (2006) Use of composted sewage sludge in growth media for broccoli. Bioresource Technol 97: 123-30. Pierzynski G M and Schwab A P (1993) Bioavailability of zinc, cadmium and lead in a metal contaminated alluvial soil. J Environ Qual 22: 247-54. Pinto A P, Mota A M, DeVarennes A and Pinto F C (2004) Influence of organic matter on the uptake of cadmium, zinc, copper and iron by sorghum plants. Sci Total Environ 326: 239-47. Piper C S (1966) Soil and plant analysis. Hans Publishers, Mumbai.

82

Prabu P C (2009) Impact of heavy metal contamination of Akaki river of Ethiopia on soil and metal toxicity on cultivated vegetable crops. Electronic J Environ Agric Food Chem 8: 818-27. Puri A N (1949) A new method of estimating carbonates in soils. Bull imp Agri Res. Pusa 206.p7. Putwattana N,Kruatrachue M, Pokethitiyook P and Chaiyarat R (2010) Immobilization of cadmium in soil by cow manure and silicate fertilizer, and reduced accumulation of cadmium in sweet basil (Ocimum basilicum). Sci Asia 36: 34954. Raicevic S, Kaludjerovic-Radoicic T and Zouboulis A I (2005) In situ stabilization of toxic metals in polluted soils using phosphates: theoretical prediction and experiment verification. J Hazard Mater 117: 4153. Ram N and Verloo M (1985) Influence of organic material on the uptake of heavy metals by corn in a polluted Belgian soil. Pedologie 35: 147-53. Ramos I, Hernandezi L M and Gonzaliz M J (1994) Sequential fractionation of copper, lead, cadmium and zinc in soils near Donana national park. J Environ Qual 23: 50-57. Rana R P and Kansal B D (1985) Micronutrient content in berseem ( Trifoliumalexandrium) as affected by the rate of cadmium application in soils. Ind J Pl Nut 3: 287-91. Rattan R K, Datta S P, Chandra S and Saharan N (2002) Heavy metals and environmental quality. Ferti News 47: 21-26 Rattan R K, Datta S P, Chhonkar P K, Suribabua K and Singh A K (2005) Long-term impact of irrigation with sewage effluents on heavy metal content in soils, crops and ground water a case study. Agric Ecosys Environ 103 (3-4): 310-22. Richards L A (1954) Diagnosis and improvement of saline and alkali soils. USDA Hand Book No 60, Washington, DC. Romero-Puertas M C, Corpasa F C Rodriguez M, Del Rio L A, Sandalio L M (2002) Cadmium caues the oxidative modification of protiens in pea plant. Pl Cell Environ 109: 427-33. Root R A, Miller R J, Koeppe D E (1975) Uptake of cadmium its toxicity and effect on the iron ratio in hydroponically grown corn. J Environ Qual 4: 473476. Rupp D, Ruhl E, Alleweldt G (1985) Cadmium toxicity in grape vines. Vitis 24: 8896. Rusan M J M, Hinnawi S and Rousan L (2007) Long term effect of waste water irrigation of forage crops on soil and plant quality parameters. Desalinization 215: 143-52. Sadamoto H, Limura K, Honna T and Yamamoto S (1995) Examination of fractionation of heavy metals in soils. Jap J Soil Sci Pl Nutr 65: 645-53 (Original not seen, Abstr in CAB Abstract A.N 951904498). Sadana U S and Singh B (1987) Effect of zinc application on yield and cadmium content of spinach grown on cadmium polluted soils. Anal Biol 3: 59-60. 83

Saini S P and Kansal B D (1990) Abatement of toxicity of sewage water and industrial effluents in maize fodder. J Pl Sci Res 6: 55-59. Sandalio L M, Dalurzo H C, Gomej M Romero- Puertas M C and Del Rio L A (2001) Cadmium induced changes in growth and oxidative metabolism of pea plants. J Exp Bot52: 2015-26. Sanita di Toppi L and Gabbrielli (1999) Response to cadmium in higher plants. Environ Exp Bot 41: 105-30. Shafiq M, Zafar M and Athar M (2008) Effect of lead and cadmium on germination and seedling growth of Leucaena leucocephala. J Appl Sci Environ Manage 12(2): 61-66. Sharma V K and Kansal B D (1986) Heavy metals contamination of soils and plants with sewage irrigation. Pollun Res 4: 86-91. Shuman L M (1982) Sepratior of soil Iron and manganeseoxide fractions from micro elementsanalysis. Soil Sci Soc Am J 46: 1099-1102. Shuman L M (1983) Sodium hyperchlorite methods for extracting microelements associated with soil organic matter. Soil Sci Soc Am J 47:656-60. Shuman L M (1985) Fractionation method for soil microelements. Soil Sci 140: 11-12. Shuman L M (1988) Effect of organic waste amendments on cadmium and lead in soil fraction of two soils. Commum Soil Sci Pl Anal 29: 2939-52. Sidhu V P S and Khurana M P S (2010) Effect of cadmium-contaminated soils on dry matter yield and mineral composition of raya (brassica juncea) and spinach (spinaciaoleracea) Acta Agronomica Hungarica, 58(4) doi: 10.1556/aagr. 58. 2010. 4 Siebe C (1998) Nutrient inputs to soil and their uptake by alfalfa through long-term irrigation with untreated sewage effluent in Mexico, Soil Use Manage, CAB International Oxford, 1998, pp. 119-122 Sikka R (2003) Influence of soil characteristics on available Pb and Cd and their accumulation by Brassica Spp. Ph.D. dissertation. Punjab Agricultural University, Ludhiana, India Silviera D J and Sommers L E (1977) Extractability of copper, zinc, cadmium and lead in soils incubated with sewage sludge. J Environ Qual., 6, 47-52. Singh A, Sharma R K, Agrawal M and Marshall F M (2010) Risk assessment of heavy metal toxicity through contaminated vegetables from waste water irrigated area of Varanasi, India. Tropical Ecol 51: 375-87. Singh B R, Narwal R P, Jeng A and Almas A (1995) Crop uptake and extractability of cadmium in soils naturally high in metals at different pH values. Commum Soil Sci Pl Anal l26: 2123-42. Singh B, Kumar V, Antil R S, and Ahlawat V S (1992) Cadmium intake by wheat as influenced by nitrogen and FYM application in sandy soil. Crop Res 5: 24348. 84

Singh E. R and Steinnes E (1976) Uptake of trace elements by barley in Znpolluted soils. II. Lead, cadmium, mercury, selenium, arsenic, chromium and vanadium in barley. Soil Sci 121: 38-43. Singh J and Kansal B D (1985) Effect of long term application of municipal waste on some chemical properties of soils. J Res Punjab Agri Uni 22: 235-42. Singh J P, Karwasra S P S and Singh M (1988) Distribution and forms of copper, iron, manganese and zinc in calcareous soils of India. Soil Sci 146(5): 359-66. Singh S and Aggarwal P K (2005) Effect of heavy metal fertilizer on growth yield distribution in wheat. Ind J Pl Physiol 10: 302-05. Singh S P and Nayyar V K (1991) Effect of cadmium on growth and cadmium and zinc content of wheat on a typic-ustepsamments. J Ind Soc Soil Sci 39: 204-05. Singh S P and Nayyer V K (1989) Effect of cadmium and zinc on growth of corn in a coarse texture soil (TypicUstipsamments). Int J Environ Studies 34: 57-63. Singh S P and Nayyer V K (1993) Adsorption of cadmium on alkaline and calcareous soils. Ind Soc Soil Sci 41: 271-73. Singh S P and Nayyer V K (2001) Influence of lime on nickel availability to plants and its toxic level in cowpea. J Res Punjab Agri Uni 30: 10-13. Singh S P, Nayyar V K (1989): Accumulation characteristics of cadmium and its upper criticallevels in selected vegetable species. Int J Environ Studies 36:199204. Singh S P, Nayyar V K (1994): Accumulation characteristics of cadmium in selected forage species. J. Ind Soc. Soil Sci., 42, 96100 Singh S P, Tack F M and Verloo M G (1998) Heavy metal fractionation and extractability in dredged sediments derived surface soils. Water, Air and Soil Pollut 102: 313-18. (Original not seen, Abstr in CAB Abstract A.N 951908256). Soon Y K (1981) Solubility and sorption of cadmium in soils amended with sewage sludge. J Soil Sci 32: 85-95. Soon Y K and Bates T E (1982) Chemical pools of cadmium, nickel and zinc in polluted soils and some preliminary indications of their availability to plants. J Soil Sci 33: 477-88. Sposito G, Lund L J and Chang A C (1982) Trace metal chemistry in arid zone field soils amended with sewage 1.Fractionation of Ni, Zn, Cu, Cd and Pb in solid phases. Soil Sci Soc Am J 46: 260-64. Stover R C Sommers L E and Silviera D J (1976) Evaluation of metals in waste water sludge. J Water Pollut Control Fed 48: 2165-75. Street J J, Sabey B R and Lindsay W L (1978) Influence of pH, phosphorus, cadmium, sewage sludge and incubation time on the solubility and plant uptake of cadmium. J Environ Qual 7: 286-90.

85

Tayawade S S and Prasad J (2008) Characterization of sewage-water-irrigated and nonirrigated soils in Nag River ecosystem of Nagpur, Maharashtra. J Ind soc soil sci 56: 247-53 Taylor R W, Xiu H, Mehadi A A, Shuford J W and Tadesse W (1995) Fractionation of residual cadmium, copper, nickel, lead and zinc in previously sludge amended soil. Commum Soil Sci Pl Anal 26: 2193-204 (Original not seen, Abstr in CAB Abstract A.N 951909241). Tessier A, Campbell P G C and Bission M (1979) Sequential extraction procedure for the speciation of particulate trace metals. Anal Chem 51: 844-51. Thakur N P and Kansal B D (1992) Effect of cadmium application to soils on dry matter yield and Cd concentration in maize fodder. Ind J Ecol 19: 15-19. Tlustos P, Szakova J, Korinek K , Pavlikova D, Hancb A and Balik J (2008) The effect of liming on cadmium, lead, and zinc uptake reduction by spring wheat grown in contaminated soil. Pl Soil Environ 52: 1624. Valsami-Jones E, Ragnarsdotteir K V, Putnis A, Bosbach D, Kemp A J and Cressy G (1998) The dissolution of apatite in the presence of aqueous metal cations at pH 2-7.Chem Geol 151: 215-33. Van Lune, P, Zwart, K B (1997) Cadmium uptake by crops from the subsoil. Pl Soil 189: 231-37. Wagner G J (1993) Accumulation of cadmium in crop plants and its consequences to human health. Adv Agron 51: 172-212. Walker W M, Boggers S F and Hassett J J (1979) Effect of cultivar and Cd rate upon the nutrient content of soybean plants. J Pl Nutr 1: 273-89. White M C and Chaney R L (1980). Zinc Cd and Mn uptake by soybean from Zn and Cd amended coastal plain soils. Soil Sci Soc Am J 44 : 308-13. Williams D E, Vlamis J, Pukite A H and Corey J E (1980) Trace elements accumulation, movement and distribution in the soil profile from massive application of sewage sludge. Soil Sci 129: 119-32. Xian X (1989b) Effect of chemical forms of cadmium, zinc and lead in polluted soils on their uptake by cabbage plants. Pl Soil 113: 257-64. Xian X and Shokohifard G I (1989a) Effect of pH on chemical forms and plant availability of cadmium, zinc and lead in polluted soils. Water Air Soil Pollut 45: 265-73. Yang M G, Lin X Y and Yang X E (1998) Impact of Cd on growth and nutrient accumulation of different plant species. Chin J Appl Ecol 19: 89-94. Zhang Y L, Dai J L, Wang R Q and, Zhang J (2008) Effects of long-term sewage

irrigation on agricultural soil microbial structural and functional characterizations in Shandong, China. European J Soil Biol 44 (1): 84-91.
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Zhu Y G, Chen S B and Yang J C (2004) Effects of soil amendments on lead uptake by two vegetable crops from a lead-contaminated soil from Anhui, China. Environ Int 30: 35160. Zwonitzer J C, Pierzynski G M and Hettiarachchi G M (2003) Effects of phosphorus additions on lead, cadmium and zinc bioavailabilities in a metal-contaminated soil. Water Air Soil Pollut 143: 193209.

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VITA
Name of the student Fathers name Mothers name Nationality Date of Birth Permanent home address : : : : : : Dharamvir Singh Kambo Sh. Amarjit Singh Smt. Balwinder Kaur Indian 28th July, 1986 VPO Phulewala. Teh. Bagha purana Distt. Moga

Educational Qualification Bachelors degree University Year of award % of marks Masters degree OCPA Year of award Title of Masters Thesis : : : : : : : : B.Sc (Ag.) Hons Punjabi University, Patiala 2008 64.7% M.Sc. (Soils) 7.58/10 2011 Delineation of cadmium contaminated soils around Buddah Nallah (Ludhiana) and remedial measures of affected soils MET Scholarship (2008)

Awards and distinctions

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