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Ecological Engineering 71 (2014) 318–325

Contents lists available at ScienceDirect

Ecological Engineering

journa l h om epa ge: www.elsev ier .com/ locate /eco leng

owards the ecological profiling of a pesticide contaminated soil siteor remediation and management

ishal Tripathia, Rama Kant Dubeya, Sheikh Adil Edrisi a, Kamini Narainb, H.B. Singhc,andita Singhd, P.C. Abhilasha,∗

Institute of Environment & Sustainable Development, Banaras Hindu University, Varanasi 221005, IndiaSchool of Environmental Science, Babasaheb Bhimrao Ambedkar (Central) University, Lucknow, IndiaDept. of Mycology & Plant Pathology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi 221005, IndiaEco-Auditing Group, CSIR-National Botanical Research Institute, Lucknow 226001, India

r t i c l e i n f o

rticle history:eceived 10 April 2014eceived in revised form 23 June 2014ccepted 15 July 2014

eywords:cological profilingexachlorocyclohexane isomerslant diversityicrobial Biomassermination assaysensitive species

a b s t r a c t

Ecological profiling of a contaminated system is the first and foremost requirement for any kind of in siturestoration activities. Therefore, in the present study, we sought to validate simple, adequate and consis-tent inventorying and monitoring methods for the ecological characterization of a hexachlorocylcohexane(HCH) isomers contaminated soil site in Lucknow, North India. The abundance and diversity of plantspecies, microbial biomass, total organic carbon, soil dehydrogenase activity, pesticide concentration insoil and plant species as well as the occurrence of ecologically sensitive species such as earthworms,honey bees and butterflies in contaminated and non-contaminated soil sites were studied. FTIR analysiswas done for assessing the variation of functional groups present in soil. Furthermore, the germinationassays of selected seeds were conducted in both HCH-contaminated and non-contaminated soils. Interest-ingly, there was a significant difference (p ≤ 0.01) observed for the studied variables in HCH-contaminatedsite in comparison to the non-contaminated soil site and they were negatively correlated with the levelof HCH contamination. Twenty five plant species were reported from the control site; whereas in thecase of HCH-contaminated site, it was reduced to seven species. The presence of �-, �-, �- and �-HCHisomers in the soil samples of contaminated sites were varied from 5.18–12.45, 30.15–68.77, 6.93–16.55and 0.75–7.54 mg kg−1, respectively, whereas the concentrations of

∑HCH in plant samples were varied

from 2.78 to 12.47 mg kg−1. The germination percentages of all the test plants were significantly low in

contaminated soil. Most interestingly, honeybees, earthworms and butterflies were not spotted in thecontaminated sites. The study indicates that proposed way of ecological characterization is appropriatefor (i) knowing the extent and level of HCH isomers contamination (ii) knowing the adaptive capacityof the contaminated soil system and (iii) for adopting suitable methodical frame works for the in siturestoration of contaminated soil sites.

© 2014 Elsevier B.V. All rights reserved.

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. Introduction

Soil is an important life supporting system of the planetarth. However, the contamination of soil system due to variousnthropogenic activities is one of the most serious problemsf environmental pollution worldwide (Abhilash et al., 2012;

olchko et al., 2014). The rapidly growing human population exerts

remendous pressure on land (Godfray et al., 2010) and it has beenstimated that around 30% of the global land is degraded due to

∗ Corresponding author. Tel.: +91 9415644280.E-mail addresses: pcabhilash@hotmail.com, pca.iesd@bhu.ac.in (P.C. Abhilash).

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ttp://dx.doi.org/10.1016/j.ecoleng.2014.07.059925-8574/© 2014 Elsevier B.V. All rights reserved.

arious kind of anthropogenic activities including contaminationue to heavy metals and persistent organic pollutants (POPs)Banwart, 2011; Abhilash et al., 2012, 2013a). Unfortunately, theumber of contaminated sites is consistently increasing all overhe world (Huttermann et al., 2009; Banwart, 2011; Ma et al., 2011;bhilash et al., 2012). These contaminated sites pose a serious

hreat to human health and environment thus requires immediatettention for revitalization. Although the developed countriesave framed suitable methodologies for the ecotoxicological risk

rofiling of POPs contaminated sites (US Environmental Protectiongency (US EPA), 1989, 1992; Swartjes et al., 2008; Kiel, 2013)nd declared such sites as ‘superfund sites’ (Anderson et al., 2002;u and Falk, 2002; Charnley and Engelbert, 2005; Harte et al.,

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012; Walker et al., 2013) or ‘brown fields’ (De Sousa, 2002;dams et al., 2010; Dixon et al., 2011; Wernstedt et al., 2013) forontainment and management, such efforts are still lacking ineveloping countries including India. Moreover, lack of methodicalrameworks for the ecological characterization of hazardous siteselays the implementation of appropriate site specific remedia-ion strategies for the onsite management of contaminated sitesAbhilash et al., 2013a). In this context, ecological profiling and soilealth assessment are the key parameters for the identification of

contaminated site for its proper restoration and management.ost importantly, such assessments would provide an idea about

he adaptive capacity of the contaminated soil system (Abhilasht al., 2013b).

Therefore, the present study was aimed to develop a costffective ecological profiling method for the screening and char-cterization of a pesticide contaminated soil site in Lucknow,orth India for remediation and management. For this purpose,

wo different soil sites of contrasting features were selected. Theontaminated site selected for this study was adjacent to a lin-ane (1,2,3,4,5,6-hexachorocycohexane; �-HCH) producing factoryIndia Pesticide Limited (IPL) situated at Chinhat Small Scale Indus-rial area nearby to Lucknow city in Uttar Pradesh, India wheres the control site was situated about 8 km away from the con-aminated site. The contaminated area is located between 27◦55′

orth latitude and 89◦3′ east longitude. The industry is functionalince 1993 and is producing lindane in pure and technical formula-ions. The production of lindane generates large quantities of wasteCH isomers such as �-, �- and �-HCH. Although these isomers doot possess insecticidal properties, they are persistent, hazardousnd reported to have high mammalian toxicity. As a result, �-,- and �-HCH were recently included in the Annexure-A of thetockholm Convention on POPs (Vijgen et al., 2011) for global elim-nation. However, its restricted use is still permitted to India forombating malaria (Vijgen et al., 2011). Although the toxic effectsf �-, �- and �-HCH are widely known to scientific community, stillhere is paucity of scientific knowledge on the combined effectsf these pollutants on biological organization such as populations,ommunities and ecosystem (Köhler and Triebskorn, 2013). There-ore, proper measures should be taken to prevent the detrimentalmpacts pose by contaminated sites and such sites should be prop-rly characterized and demarked for further necessary action. Inhis context, an ecological profiling of a lindane and other wasteCH isomers contaminated soil has been done for decision makingrocesses and developing onsite remediation strategy.

. Materials and methods

Ecological profiling was conducted in a pesticide contaminatedoil site in the premises of a lindane producing factory named IPLt Chinhat Industrial Estate, Lucknow, North India and the compar-son was made with a control site located about 8 km away fromhe pesticide contaminated site (Fig. 1). Both the sites were ana-ysed for the occurrence of key ecological indicators such as (i)iological indicators (ii) soil quality indicators and (iii) pollution

ndicators. Frequent field visits (during the morning and eveningours) were conducted to record the presence or absence of eco-

ogically sensitive species such as honey bees and butterflies inoth contaminated and controlled sites. In order to notice the pres-nce of earth worms, the soils were randomly dug up to 60 cm andecorded their presence or absence. The diversity and abundancef the plant species were recorded by quadrate method (25 × 25 m)

nd the species identity were confirmed by using ‘Flora of India’.he leave samples of the abundant species growing at the con-aminated and controlled sites were collected for analysing theoncentration of HCH isomers.

eicC

eering 71 (2014) 318–325 319

The sub-surface soil samples were collected (10–15 cm) in trip-icate from different locations of the contaminated and control soilites. The fresh soil samples were analyzed for pH (pH meter; Cybercan-500), EC (EC meter, Cyber Scan-500), total organic carbonYeomansa and Bremnera, 1988) and microbial biomass carbonVances et al., 1987). Soil dehydrogenase activity was analysedy monitoring the rate of reduction of 2 3,5-triphenyltetrazoliumhloride (TTC) to the red, water insoluble triphenylformazan (TPF)Fan et al., 2008). Bacterial isolation and the enumeration of colonyorming units (CFU) were carried out from the soil samples of bothontaminated as well as control sites in accordance of standardrocedures published earlier (Sahu et al., 1995; Nagata et al., 1999;urthy and Manomani, 2007; Elcey and Kunhi, 2010; Abhilash

t al., 2011). A portion of the soil samples were air dried and passedhrough a 2 mm sieve and stored at −18 ◦C for pesticide residuenalysis.

Analysis of �-, �-, �- and �-HCH isomers from soil samplesere performed through soxhlet extraction followed by GC-ECD

Abhilash and Singh, 2008). Similarly, plant samples were extractedy matrix-solid phase dispersion extraction (MSPD) and the HCH

somers concentration in the extracts were determined by gas-hromatograph equipped with 63Ni-electron capture detector. Theample preparation, method development, analytical, instrumen-al, calibration, quality assurance and quality control protocols forCH isomers extraction and analysis from soil and plant samplesere detailed in our earlier publications (Abhilash et al., 2007,

009, 2011; Abhilash and Singh, 2008).Fourier transform infrared (FTIR) spectroscopic measurements

ere conducted to examine the effect of HCH isomers contam-nation on the chemical functional groups present in soil. Forhis, soil samples were characterized by infrared spectroscopy (IR)sing FTIR (NicoletTM 6700, Thermo Scientific, USA). One mg ofoil sample was finely grounded with hundred mg of potassiumromide (KBr) and the mixture was pelletized using a hydraulicump named CAP-15T. The spectra of soil samples were obtainedt 4000–400 cm−1 mid-IR range (Narain et al., 2012). In order totudy the phytotoxicity of contaminated soils, germination assayas performed for the seeds of Vigna radiata(L.) R. Wilczek, Jatropha

urcas L. and Sesamum indicum L.. For this, 100 seeds of each of thelant species were grown in contaminated as well as control soil.ermination was observed and germination percentage was cal-ulated by observing the number of seeds germinated out of 100eeds of each plant species.

The data were subjected to analysis of variance (ANOVA) bysing SPSS-16.0 (Illinois, USA) software for windows program. Theistribution of various HCH isomers was presented using box andhisker plot. The effects of HCH contamination on plant diver-

ity, microbial biomass, soil quality etc were compared by linearegression and Pearson coefficient of correlation.

. Results and discussion

.1. Soil quality indicators

The soil quality indicators of both pesticides contaminated andhe control sites are presented in Table 1. Soil microbes (expresseds CFU g−1 soil), microbial biomass carbon (MBC), total organicarbon (TOC) and soil dehydrogenase activity were taken as theey indicators for assessing the soil quality and soil health. Asrom Table 1, it is apparent that there was a significant differ-

nce observed in the correspondent values of the soil qualityndicators of the contaminated soil site in comparison to the non-ontaminated (control) site at 99.9% confidence level ( ̨ = 0.01). TheFU count of the contaminated site was found to be 5–6 times lower

320 V. Tripathi et al. / Ecological Engineering 71 (2014) 318–325

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ig. 1. The methodology employed for the ecotoxicological profiling of a HCHs conlant diversity, concentration of HCH in soil and leaves, germination assays etc. for

han the control site which clearly indicates the negative effect ofCH isomers on microbial diversity in soil. This was further evident

rom the values of MBC content in contaminated (45–64 �g kg−1)nd control sites (150–200 �g kg−1). Furthermore, both MBC andFU counts were negatively correlated (r2 = −0.89 and r2 = −0.91,espectively) with the level of HCH contamination in soil (please seeection 3.2). The microbial biomass is the key driver of soil organicatter decomposition and synthesis (Niemeyer et al., 2012) and

t can be used to define the impact of soil pollution on micro-ial activities of the soil (Zhou et al., 2012; Jacobson and Hjelmso,014). Therefore, the lower level of organic carbon in the con-aminated site (0.05–0.15%) might be attributed due to the lower

icrobial biomass in the contaminated area. Similar to the abovendings, there was a marked difference (p < 0.001) observed inhe case of the soil dehydrogenase activity of both the contami-ated and control sites. Soil dehydrogenase activity was about 3imes higher in non-contaminated soil than HCH-contaminatedoil. Soil dehydrogenase activity is a useful index for measuringhe dehydrogenation of organic matter by transferring electronsrom substrates to acceptors and can be considered as one of the

ey microbial indicators of soil fertility (Waksman, 1992). Reducedehydrogenase activity in HCH contaminated site shows the lowerxidative metabolism of microorganisms in stressed ecosystemVelmourougane et al., 2013; Pandey et al., 2006.) Similarly, Baruah

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ated soil site. We have used certain soil quality parameters, biological indicators,ological characterization of the study site.

nd Mishra (1986) also reported the reduced dehydrogenase activ-ty due to the higher doses of pesticides such as butachlor, 2,4-d andxyfluorfen. The IR spectra of the HCH contaminated and controlites along with the correspondent functional groups are depictedn Fig. 2. The distinct difference in the spectral characteristics ofhe contaminated and non-contaminated soil sites clearly indi-ates the presence or absence of various chemical functional groupsresent in the soil (Fig. 2). The above findings clearly indicate thatoil contamination due to HCH isomers can alter the functionalitynd health of the soil system by disturbing its physicochemical andicrobiological properties.

.2. Soil pollution indicators

The concentration of HCH-isomers in soil has been directlyaken as a measure to index the soil pollution in the area (Fig. 3).ll the major HCH isomers (�, �-, �- and �-HCH) were present in

he contaminated area; however, only �-HCH (0.15 to 0.3 mg kg−1)as reported from the control site. Among the four isomers, �-CH was found in higher amount (30 to 69 mg kg−1) followed by

-HCH (7 to 17 mg kg−1), �-HCH (5–12 mg kg−1) and �-HCH (0.75o 8 mg kg−1). The higher level of �-HCH in soil is mainly due tots chemical stability against microbial degradation. Most impor-antly, the higher level of �-HCH in soil samples may be attributed

V. Tripathi et al. / Ecological Engineering 71 (2014) 318–325 321

Table 1Ecotoxicological profiling of the contaminated soil site in comparison to a control site using soil quality, plant diversity and other biological indicators.

Sl. No Variables Values

HCH-contaminated Site Non-contaminated site

(A) Soil quality indicators (n = 5)1 Soil microbes (CFU g−1 soil) 3 × 106–5 × 106 18 × 106–23 × 106

2 Microbial biomass carbon (�g kg−1) 45–64 150–2003 Total organic carbon (%) 0.05–0.15 0.35–0.514 Soil dehydrogenase (�g TPF g−1 dwt soil) 12–18 40–55(B) Biological indicators*

1 Presence of earth worms Not found ++2 Presence of honey bees/bees Not found +++3 Presence of butterflies Not found +(C) HCHs concentration in soil samples (mg kg−1 samples) (n = 5)1 �-HCH 5.18–12.45 ND2 �-HCH 30.15–68.77 ND3 �-HCH 6.93–16.55 0.15–0.284 �-HCH 0.75–7.54 ND(D) Phytoaccumulation of

∑HCH** in leaf samples (mg kg−1 samples) (n = 3)

1 Achyranthes aspera L. 5.75 ± 0.25 ND2 Calotropis procera (Ait). Ait 4.83 ± 0.30 ND3 Dalbergia sisso Roxb. 9.45 ± 0.11 0.28 ± 0.024 Erianthus munja (Roxb.) Jesw. 2.78 ± 0.05 ND5 Lantana camara L. 6.09 ± 0.12 ND6 Solanum torvum L. 10.88 ± 0.15 ND7 Withania somnifera (L.) Dunal 12.47 ± 0.20 ND(E) Germination assays (n = 5)1 Vigna radiata (L.) R. Wilczek 23–36% 96–98%2 Jatropha curcas L. 27–43% 94–97%3 Sesamum indicum L. 18–25% 92–96%

* The symbols ‘+’, ‘++’ and ‘+++’ indicates ‘low’, ‘medium’ and ‘high’ level of abundance.**

∑HCH (sum of �-, �-, �- and �-HCH).

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Fig. 2. The FTIR spectra of the soil samples from contaminated and control soil sites. The wave numbers (cm−1) and their respective functional groups are: 3908-3623: hydroxylOH stretching, phenols, alcohol, N H containing amine or amide, or carboxylic acid; 3290-3273:O H and N H stretching vibration (amino acids); 3156: aromatic C H

stretch; 2960: CH, CH2 and CH3 carbon to hydrogen stretching vibration; 2937-2920: methylene C H asym./sym. stretch, aliphatic C H stretch; 2401-2228: CC and CNtriple bond (nitriles); 1712: C O of COOH, C O or ketonic carbonyl; 1681-1613: C O stretching of amide groups (amide I) quinine, C O stretching of H-bonded conjugatedketones; 1563-1515: COO− symmetric stretching, N H formation and C N stretching (amide II band), C C of aromatic rings; 1459-1400: methylene C H bend; aliphaticC H; 1417-1399: aromatic ring stretch and COO− anti-symmetric stretching; 1239-1231: C O stretch of OH-deformation of COOH; 1153-1104:S O in sulfone groups;1072-1023: carboxylic acids, esters, ethers, alcohols and anhydrides; 999-1024: C O stretching of polysaccharides and Si O asymmetric stretch; 851-832: hydrogenbonded OH-deformation in carboxyl groups; 776-688 methylene-(CH2)n-rocking (n ≥ 3) C H bending vibrations, Si O Si stretching vibration; 526-426: C O O, P O Cbonding (aromatics) phosphates.

322 V. Tripathi et al. / Ecological Engin

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ig. 3. Distribution of HCH isomers (�-, �-, �- and �-HCH) in contaminated soil site.

y the isomerization of �-HCH to �-HCH and of �-HCH via �-HCHo the more stable �-HCH (Fiedler et al., 1995; Wu et al., 1997;

anz et al., 2001; Abhilash and Singh, 2008). The occurrence of �-CH (lindane) in soil samples indicate the contamination due to

pill over during the production, storage and transportation pro-esses. Therefore, the level of �-HCH can be directly taken as a keyndicator of HCH pollution in soil (Abhilash and Singh, 2008).

.3. Biological indicators

Biodiversity is a key indicator of landscape sustainabilityPaoletti, 1999). The diversity and abundance of plant species grow-ng at the contaminated and control sites are presented in Table 2.here were 25 species of plants growing at the controlled sites,hereas in the case of HCH-contaminated soil sites, the diversityas reduced to 7. The habit of the most of the plants growing in

he contaminated area of India pesticide limited were herb andhrub, however, 12 tree species were reported from the controlite (Table 2). Alstonia scholaris (L.) R.Br., Leucaena leucocephalaLam.) de Wit, Mangifera indica L., Pongamia pinnata (L.) Pierre andolyalthia longifolia (Sonn.) Hook. F. & Thomson are the most com-only found tree species in the control site, whereas Saccharumunja was the most abundant species reported at the contaminated

ite. However, it was not abundant at the control site. Previoustudies also reported that S. munja can colonize in extreme habi-ats (Pandey et al., 2012; Rau et al., 2009). Its ability to produce highiomass, profuse root network to tightly bind the soil particles arehe key features favouring its rapid growth and survival in harshnvironments (Sharma et al., 2011). Thus native plant species ofhe polluted sites such as Solanum nigrum, S. munja could be useds indicator of stressed soil and may be exploited for revitalizationf such degraded lands.

The diversity and abundance of all plant species were report-dly higher at the control site in comparison to the contaminatedite (Table 2). Similar observations were also reported in earliertudies. Schipper et al. (2011) studied the plant species variationnd richness in Rhine river floodplain in the context of heavy metalontamination of soil (Schipper et al., 2011) and the study found

hat the species diversity and richness was decreased due to theeavy metal contamination in the studied soil. Similarly, Struckhofft al. (2013) reported that the soil pollution itself can adverselyffect the native floristic quality. Furthermore, a reduction in plant

Bihs

eering 71 (2014) 318–325

iversity and richness was observed in an open waste dumpingazardous site in Islamabad city in Pakistan. Only 32 plant speciesere reported at the disposal site in comparison to 44 plant species

f the control site (Ali et al., 2014). In concurrence of the above find-ngs, there was a strong negative correlation was found betweenhe diversity and abundance of plant species with the CFU countnd MBC content in soils. It may be attributed by the fact thaticroorganisms also support other forms of life including plants

y providing nutrients and playing key role in cycling and syn-hesis of various micro, macronutrients in soil. Furthermore, itlso alleviates the toxicity of xenobiotic compounds present in soily biodegradation (Jacobson and Hjelmso, 2014; Berendsen et al.,012; María et al., 2010). Apart from the presumed toxicity of HCH-

somers to the plant species, the lower level of microbial activityight be another factor affecting the diversity and abundance of

lant species at contaminated site. However, more studies per-aining to the plant specific microbial assemblages are essentialo validate such claims.

The accumulation of∑

HCH (sum of �-, �-, �- and �-HCH)re presented in Table 1. Among the various plant species,ithania somnifera (L.) Dunal exhibited maximum accumula-

ion (12.47 mg kg−1) followed by Solaun torvum L. (10.88 mg kg−1)hereas, the Erianthus munja (Roxb.) Jesw showed the minimum

2.78 mg kg−1) accumulation of HCH siomers. Since W. somniferand S. torvum can accumulate considerable amount of HCH iso-ers, they could be used for the onsite remediation of HCH isomers

ontaminated soil sites.Another possibility for the low level of plant diversity in con-

aminated soil may be due to the phytotoxicity of HCH isomers.t has been reported that pollutants can interfere with the var-ous plant processes such as reproduction, photosynthesis andtomatal functioning and even damaging the biological membranesBoutin et al., 2014; Dazy et al., 2009). Therefore, the diversitynd abundance of the plant species growing in the contaminatedoil sites depends up on their ability to accumulate and degradehe isomers in their tissues or their ability to exclude or avoidhe HCH-isomers (Austruy et al., 2013). Austruy et al. (2013)eported that native plants of metalliferrous contaminated soiluch as Agrostis capillaris, S. nigrum show adaptive tolerance tohe heavy metal pollution in soil. Germination assays of selectedlant species were conducted in HCH-contaminated soil sites inrder to validate the phytotoxicity of HCH-contaminated soils. Inhe present study, we found that HCH-contaminated soil signifi-antly reduce the percentage germination of test plants (p < 0.001)iz. S. indicum, J. curcas and V. radiata, whereas in the case of con-rol site (non-contaminated soil), the germination percentage wasound >95%33.

It is presumable that the higher concentration of∑

HCH (Fig. 3)n the contaminated area has degraded the land quality by alteringhe physicochemical and biochemical properties of the soil. More-ver, the contamination of HCH isomers has not only affected theicrobial activity and biochemistry of the contaminated soil site

ut also produced adverse effect on sensitive ecological indicatorpecies. As a result, not a single earthworm, its burrow or cast-ng was reported from the contaminated site however, they werebundant at control site. Earthworms are true representative ofoil heath as they are highly sensitive to soil pollution (Garcia et al.,011; Pelosi et al., 2013). Similar trend was also observed in the casef honeybees and butterflies as they were not reported from theontaminated site. Species like honeybees and butterflies have ear-ier been reported as health indicator of ecosystems (Paoletti, 1999;

adiou-Bénéteau et al., 2013). Carvalho et al. (2013) reported that

nsecticides deltamethrin, fipronil, and spinosad are toxic to theoneybeeApis mellifera and therefore, it could be used as biomarkerpecies for assessing the presence of insecticides in soil. Therefore,

V. Tripathi et al. / Ecological Engineering 71 (2014) 318–325 323

Table 2Diversity and abundance of plant species in contaminated and control site.

Sl. No Scientific Name Family Habit Abundance*

Contaminated site Control site

1 Achyranthes aspera L. Amaranthaceae Herb + ++2 Albizia lebbeck (L.) Benth Mimosaceae Tree Not found +3 Alstonia scholaris (L.) R.Br. Apcocynaceae Tree Not found ++4 Annona squamosa L. Annonaceae Shrub Not found +5 Azadirachta indica A. Juss Meliaceae Tree Not found +6 Calotropis procera (Ait). Ait Asclepadaceae Shrub + +7 Cynodon dactylon (L). Pers. Poaceae Herb Not found +++8 Dalbergia sisso Roxb. Fabaceae Tree + +++9 Datura stramonium L. Solanaceae Herb Not found ++10 Delonix regia (Bojer) Raf. Fabaceae Tree Not found +11 Erianthus munja (Roxb.) Jesw. Poaceae Herb +++ +12 Euphorbia hirta L. Euphorbiaceae Herb Not found ++13 Ficus religiosa L. Moraceae Tree Not found +14 Lantana camara L. Verbenaceae Shrub ++ +15 Leucaena leucocephala (Lam.) de Wit Mimosaceae Tree Not found ++16 Mangifera indica L. Anacardiaceae Tree Not found ++17 Phyllanthus emblica L. Phyllanthaceae Tree Not found +18 Phyllanthus niruri L. Phyllanthaceae Herb Not found ++19 Pongamia pinnata (L.) Pierre Fabaceae Tree Not found ++20 Polyalthia longifolia (Sonn.) Hook. F. & Thomson Annonaceae Tree Not found ++21 Ricinus communis L. Euphorbiaceae Shrub Not found +22 Senna tora (L.) Roxb Fabaceae Herb Not found ++23 Senna siamea (Lam.) H.S. Erwin & Barneby Fabaceae Tree Not found +24 Solanum torvum L. Solanaceae Herb ++ +25 Withania somnifera (L.) Dunal Solanaceae Shrub + +

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he absence of these sensitive species from the contaminated soilslearly shows that the premises of the India Pesticide limited areeavily polluted by lindane and its waste isomers as the concentra-ion of lindane and other HCH isomers were higher at contaminatedite in comparison to the control site.

Literature provides ample evidences that the chemical contam-nants can alter the physical, chemical, biochemical and biologicalroperties of soil (Trasar-Cepeda et al., 2000; Zhou et al., 2012;

acobson and Hjelmso, 2014; Pardo et al., 2014). This directlylters biological activities in soil and adjacent environment whichn turn affect the plant and microbial diversity of the contam-nated sites. Therefore, soil enzymes, microbial biomass (Zhout al., 2012; Jacobson and Hjelmso, 2014), soil organic carbon,hemical functional groups, plant diversity (Schipper et al., 2011;truckhoff et al., 2013), presence of sensitive organisms such asutterflies (Oostermeijer and van Swaay, 1998), honeybees, earth-orms (Garcia et al., 2011; Pelosi et al., 2013; Du et al., 2014)

tc can be used as key indicators for assessing the status of theoil quality of the contaminated system (Fernandez et al., 2005a,b;hou et al., 2012; Niemeyer et al., 2012; Badiou-Bénéteau et al.,013; Du et al., 2014; Pardo et al., 2014). Soil enzymes are syn-hesized by the soil microorganisms and are true representativef soil health as they are quite sensitive to environmental pollu-ion (Gil-Sotres et al., 2000; Trasar-Cepeda et al., 2000; Jacobsonnd Hjelmso, 2014; Pardo et al., 2014). Similarly, the presence ofcological indicators such as honeybees, earthworms and butter-ies could be used as marker species for the ecological assessmentf a particular site since they are highly sensitive to environmen-al stress (Badiou-Bénéteau et al., 2013). This clearly indicates thatvaluation of total sustainability of soil natural functions could bearried out by analysing the above parameters. As these biologi-al indicators are easy to measure/identify, they can be exploited

s preferred candidates for the ecological profiling of contami-ated sites. We for the first time evaluated all these parameters forcological profiling of a pesticide contaminated site in an Indianubcontinent.

ilil

. Conclusion

Ecological characterization of the soil is necessary for deci-ion making process and developing methodical framework for theecovery of soil health and adopting onsite remediation techniques.urrent remediation processes are mainly targeted to reduce the

evel of contamination in soil and not much emphasises is placedo improve the soil quality and health. However, more holisticpproaches for soil health assessment are required for revitaliza-ion of degraded soil ecosystems. Microbiological, physicochemicalnd other biological parameters are essential for ecotoxicologi-al risk characterization of a particular contaminated soil system.nfortunately, such efforts are lacking in India. As more and more

and areas are contaminated in India due to various chemical pollut-nts, it is the need of the hour to develop simpler, cost effective, andasily reproducible method for the characterization of the degradedcosystems to develop a suitable approach for onsite restoration.ost importantly, such methods can also be easily deployed for the

nsite remediation of contaminated soil sites in other parts of theorld.

In this context, we for the first time have analysed the bioindi-ators of active (presence of sensitive ecological indicator species,bundance and richness of plant and microbes) and passive typemicrobial biomass carbon, soil enzyme assay and functional groupnalysis) altogether to develop a novel approach for the charac-erization of a HCH isomers contaminated soil sites in Lucknow,orth India, for remediation and management. The active groupf indicators like honeybees, butterflies and plant diversity are aubset of the environment thus reflects the condition of a particu-ar ecosystem. Moreover, they were negatively correlated with theevel and extent of HCH pollution in soil. Similarly, the soil qualityndicators were also negatively correlated with the HCH pollution

n soil. Since all these indicators (i.e. soil quality, biological and pol-ution indicators) could be easily interpreted and provide amplenformation about the current ecological condition of a particu-ar soil system, all these parameters could be coupled together for

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eveloping a regulatory stewardship and sustainable remediationrogramme.

cknowledgements

Financial support from Science Engineering Research Board,ovt. of India (No. SR/FT/LS-111/2011), and University Grants Com-ission (No. F.41-1110/2012 (SR)) is gratefully acknowledged. We

hankfully acknowledge the help rendered from Mr. Vishnu Murari,esearch Scholar, IESD-BHU for statistical analysis. Thanks are alsoue to the Director, CSIR-NBRI and Director, USIC-BABAU, Lucknowor providing instrumentation facilities and Director, Institute ofnvironment & Sustainable Development, Banaras Hindu Univer-ity for institutional support and encouragements.

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