Biotechonlogy

Bhupendra Koul · Pooja Taak
Biotechnological
Strategies
for Effective
Remediation of
Polluted Soils
Biotechnological Strategies for Effective
Remediation of Polluted Soils
Bhupendra Koul • Pooja Taak
Biotechnological Strategies
for Effective Remediation
of Polluted Soils
Bhupendra Koul Pooja Taak
School of Bioengineering & Biosciences School of Bioengineering & Biosciences
Lovely Professional University Lovely Professional University
Phagwara, Punjab, India Phagwara, Punjab, India
ISBN 978-981-13-2419-2 ISBN 978-981-13-2420-8 (eBook)

https://doi.org/10.1007/978-981-13-2420-8
Library of Congress Control Number: 2018958474
© Springer Nature Singapore Pte Ltd. 2018

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Preface
In Sanskrit, the classical language of India, “Mother Earth” has been revered as
“Ratna Garbha,” meaning repository of precious stones. In fact, gems, chemicals,
metals, and their derivatives are derived from the Earth. The entire environment is
personified as “Mother Nature” which is the creative and controlling force affecting
all species. Mother Nature is the source of all the biotic (flora and fauna) and abiotic
components which are being availed by man. In return, man has been ungrateful and
selfish and is therefore responsible for his own maladies. Although in his race to
achieve all the comforts, he has gained success in industrialization, automation,
transport, and health, but this has left behind a daunting legacy of land, air, water,
noise, radioactivity, and thermal pollutions. Undoubtedly, man has conquered time
and space and has reached the moon in search of a new life, but in the process, man
has ruined his own life and the environment on the planet. It is unfortunate that man
has been exploiting “Mother Nature” since he gained full cranial capacity from
prehistoric time till the modern era, but, the question, what man has given back to
Nature, remains unanswered. The fear of global warming, melting of ice caps,
floods, erosions, deforestation, bio-magnification, eutrophication, polluted rivers,
polluted air, radioactive hazards, uneven and untimely rainfall, crop failures, non-
availability of clean drinking water, spread of epidemics, various types of cancers,
and immunocompromised states are indicators of contamination and damage to our
food chain and the ecosystem. Among all the pollution types, soil pollution has
caused devastating consequences to the primary and secondary consumers both
directly and indirectly.
Although Mother Nature never deceives mankind, it is our moral duty to analyze
the losses done to the biodiversity and healthy environments in order to make the
Earth livable. It is time to devise eco-friendly strategies for sustainable remediation
of soils and ecosystems. The holistic approach of self-awareness and the advance-
ment in the eco-friendly soil remediation strategies will improve not only the indi-
vidual life but also the entire population, community, ecosystem, biome, and
biosphere.
This book focuses on the explicit information and recent advances on the avail-
able soil treatment techniques (Fig. 2.1) for the period up to 2018 and their future
v
vi Preface
prospects. Thus, the soil remediation strategies ensure environmental protection by

using natural resources for sustainable remediation which is economically and envi-
ronmentally beneficial for the society. This book is divided into nine chapters as
follows:
Chapter 1 Soil pollution: Causes and Consequences
Urbanization, modern agricultural practices, and industrialization have exacer-
bated the problem of environmental degradation in terms of air, water, and soil pol-
lution, at an alarming rate. Among all the pollution types, soil pollution has become
a matter of global concern as these factors have cumulative effect on the environ-
ment and human health through food chain contamination. The alarming situation
of the present state of soil pollution has forced the scientists to develop innovative,
refined, and reproducible strategies/technologies (in situ or ex situ) for sustainable
remediation of polluted soils. This chapter summarizes the various aspects of soil
pollution, its causes, and consequences.
Chapter 2 Ex Situ Soil Remediation Strategies
Ex situ techniques involve the treatment of contaminated soil, away from the
polluted soil region. Ex situ bioremediation can be operated in two ways which
include solid-phase bioremediation and slurry-phase bioremediation. Although
costly, these techniques are highly efficient, easy to control, and faster and have
greater potential to treat large number of contaminants from the soil. This chapter
explores various ex situ techniques such as land farming, biopile, windrow, soil
washing, composting, bioreactor, ion exchange, adsorption/absorption, pyrolysis,
and ultrasound technology.
Chapter 3 In Situ Soil Remediation Strategies
Physical methods generally involve disintegration of the contaminants by meth-
ods such as evaporation, heating, or washing. These methods are based on the prin-
ciples used for the extraction of desired metal from their respective mineral ores.
These techniques are suitable for the treatment of both inorganic and organic con-
taminants and involve low operational cost. The selection of the most suitable phys-
ical treatment depends specifically on the type of contaminated soil and type of the
contaminant(s) (i.e., organic, inorganic, or both). The present chapter discusses
various physical methods of soil remediation including physical separation, soil
flushing, volatilization, froth flotation, and thermal heating.
Chapter 4 Chemical Methods of Soil Remediation
Chemical methods include the addition of chemicals or solvents into the polluted
soil, so as to stabilize the pollutants, converting them into low toxic form and thus
diminishing the supply of contaminants to plants, water, and other ecosystems.
Since the complete soil remediation is difficult to achieve with biological methods,
the implementation of chemical techniques in tandem has gained much attention of
the scientists. This chapter explains several chemical remediation methods including
Preface vii
chemical leaching, chemical oxidation, chemical fixation, and electrokinetic

remediation.
Chapter 5 Biochar and Soil Remediation
Biochar has excellent potential to adsorb the contaminants from soil solution and
make them unavailable to organisms. The use of biochar facilitates the nutrient
availability and enhances the microbial activity, soil organic matter availability,
water retention, and crop production in soils. Although this method is simple,
robust, and suitable for many regions of the world, its economic estimations and
optimization should be taken into consideration for its large-scale application. This
chapter comprises of various processes which occur during biochar-mediated soil
treatment including carbon sequestration, nutrient exchange, water holding, adsorp-
tion/absorption, and oxidation/reduction.
Chapter 6 Soil Remediation Through Microbes
Microbes (bacteria and fungi) are adapted to cope with various unfavorable con-
ditions, including high temperature, alkalinity, and acidity. The use of microbe and
microbial consortium does facilitate bioremediation of polluted soils. Moreover, the
use of genetically modified microbes can increase the rate of microbe-mediated soil
remediation. Microbe-mediated soil remediation can take place via three methods,
i.e., bioventing, bioleaching, and bioaugmentation. This chapter discusses various
aspects of soil remediation aided by microbes.
Chapter 7 Soil Remediation Through Algae, Plants, and Animals
There is an expanding enthusiasm for deploying algal species for the remediation
of the organic (hydrocarbons) and inorganic pollutants (heavy metals) present in
soils. Phytoremediation mechanisms, although slow, have the potential to restore
the polluted soils. Although several chemical, physical, and biological methods
have been developed, phytoremediation is one of the safest green techniques for
treating the polluted sites. Some lower animals also play an important role in the
degradation of certain pollutants. This chapter explains various algal, plant, and
animal species which facilitate soil remediation and also highlights plants-
microbe(s) interactions for effective soil remediation.
Chapter 8 Nanobioremediation
Nanobioremediation is an emerging technique for remediation of polluted soils
which involves the use of biosynthetic nanoparticles. Because of the unique chemi-
cal and physical properties, nanoparticles have gained the attention of scientist to
make use of nanoparticle-mediated soil remediation. Nanoparticles are less toxic to
soil flora and enhance the microbial activity; therefore, nanobioremediation can be
promising where the other conventional remediation methods fail. This chapter
highlights the synthesis of nanoparticles from yeast, fungi, bacteria, and plant
extracts and their potential uses for remediation of contaminated soils.
viii Preface
Chapter 9 Case Studies and Future Prospects of Soil Remediation Strategies

Soil remediation strategies depend not only on the quality and quantity of the
contaminants but also the geographical conditions of the target site. For successful
soil remediation, different physical, chemical, and biological strategies can be
implemented in an integrated way. This chapter discusses various case studies
related to the implementation of remediation strategies at large scale. This chapter
also focuses on the explicit information and recent advances on the available soil
treatment techniques and their future prospects.
Phagwara, Punjab, India Bhupendra Koul

Contents
1 Soil Pollution: Causes and Consequences�� 1

1.1 Introduction�� 1
1.2 Causes of Soil Pollution�� 3
1.2.1 Natural Sources�� 4
1.2.2 Anthropogenic Sources�� 5
1.3 Harmful Effects of Soil Pollution �� 17
1.4 Remedies to Cope with Soil Pollution�� 27
1.5 Conclusions�� 28
References�� 31
2 Ex situ Soil Remediation Strategies�� 39
2.2 Land Farming�� 40
2.3 Biopile�� 40
2.4 Windrow �� 41
2.5 Composting�� 42
2.6 Soil Washing �� 43
2.7 Bioreactor �� 48
2.8 Absorption/Adsorption �� 49
2.9 Ion Exchange�� 49
2.10 Pyrolysis �� 50
2.11 Ultrasonic Technology�� 51
References�� 52
3 In Situ Soil Remediation Strategies�� 59
3.2 Physical Methods of Soil Remediation�� 60
3.2.1 Physical Separation�� 61
3.2.2 Soil Flushing �� 62
3.2.3 Volatilization �� 63
ix
x Contents
3.2.4 Froth Flotation�� 64

3.2.5 Thermal Remediation�� 65
References�� 73
4 Chemical Methods of Soil Remediation �� 77
4.2 Chemical Leaching �� 78
4.3 Chemical Fixation�� 78
4.4 Chemical Oxidation�� 79
4.5 Electrokinetic Remediation�� 80
References�� 82
5 Biochar and Soil Remediation�� 85
5.2 Carbon Sequestration�� 87
5.3 Nutrient Exchange�� 87
5.4 Water Holding�� 88
5.5 Adsorption/Absorption �� 88
5.6 Oxidation/Reduction�� 88
5.7 Biochar-Microbe Interaction in Soil �� 89
5.8 Remediation of Organic and Inorganic Contaminants
from Soils�� 91
References�� 94
6 Soil Remediation Through Microbes�� 101
6.2 Microbial Degradation of Petroleum Hydrocarbons�� 102
6.3 Bioventing�� 111
6.4 Bioleaching �� 112
6.5 Bioaugmentation�� 113
6.6 Mycoremediation�� 114
References�� 118
7 Soil Remediation Through Algae, Plants and Animals�� 129
7.2 Phycoremediation �� 130
7.3 Phytoremediation�� 134
7.3.1 Rhizofiltration �� 135
7.3.2 Phytotransformation�� 161
7.3.3 Phytoextraction/Phytoaccumulation �� 163
7.3.4 Phytomining�� 164
7.3.5 Phytostimulation �� 164
Contents xi
7.4 Genetic Engineering Approaches for Phytoremediation�� 165

7.5 Animal-Mediated Soil Remediation �� 174
8 Nanobioremediation�� 197
8.2 Properties of Nanoparticles�� 199
8.3 Types of Nanoparticles �� 200
8.3.1 TiO2 (Titanium Dioxide) Based Nanoparticles�� 200
8.3.2 Iron Based Nanoparticles�� 201
8.3.3 Bimetallic Nanoparticles�� 201
8.3.4 Nanoclays�� 203
8.3.5 Nanotubes �� 203
8.3.6 Dendrimers and Nanosponges�� 204
8.3.7 Magnetic Nanoparticles�� 204
8.4 Green Synthesis of Nanoparticles for Soil Remediation�� 205
8.5 Nanobioremediation (NBR)�� 205
9 Case Studies and Future Prospects of Soil
Remediation Strategies�� 221
9.2 Case Studies on Soil Remediation�� 222
9.2.1 Case Study 1 �� 222
9.2.2 Case Study 2 �� 223
9.2.3 Case Study 3 �� 224
9.2.4 Case Study 4 �� 224
9.2.5 Case Study 5 �� 225
9.2.6 Case Study 6 �� 226
9.2.7 Case Study 7 �� 227
9.2.8 Case Study 8 �� 228
9.2.9 Case Study 9 �� 229
9.2.10 Case Study 10 �� 231
9.2.11 Case Study 11 �� 231
9.2.12 Case Study 12 �� 233
9.2.13 Case Study 13 �� 234
9.2.14 Case Study 14 �� 235
9.2.15 Case Study 15 �� 236
9.2.16 Case Study 16 �� 236
9.3 Future Prospects�� 237
About the Authors
Dr. Bhupendra Koul is serving as an Assistant Professor in the School of

Bioengineering and Biosciences, Lovely Professional University, Punjab, India. He
was awarded with CSIR-Senior Research Fellowship in the year 2013 during his
research at Plant Transgenics Lab., CSIR-National Botanical Research Institute
(CSIR-NBRI), Lucknow. During his Ph.D., he has worked on the optimization,
introduction, and expression of modified full-length and truncated versions of
Bt-cry1Ab and Bt-cry1Ac genes in tomato for developing non-chimeric and stable
transgenic lines resistant to two lepidopteran insects, Helicoverpa armigera and
Spodoptera litura, and evaluated the performance of both the versions of cry1Ab
and cry1Ac genes for the stability and efficacy of insecticidal toxin in transgenic
plants. He has evaluated the performance and role of different cis-motifs of artificial
synthetic promoters for overexpression of genes in tomato and performed compara-
tive in silico analyses of different cry1A genes for toxicity to target insects. He has
developed several stable transgenic tomato plants resistant to plant pathogens
through the expression of protein transcription co-activator TAF4B gene and chi-
tinolytic protein toxic to whiteflies. He has 5.9 years of research experience and
4 years of teaching experience to the UG and PG students and has received the
“Teacher Appreciation Award, 2016” at LPU, in the Discipline of Biotechnology
and Biosciences, through MHRD minister, Government of India. He is well
acquainted with the techniques of plant tissue culture, gene cloning, Agrobacterium-
mediated transformation, and molecular characterization of transgenic plants
including immunological and insect bioassays. He has designed the full-length syn-
thetic cry1Ac gene (NCBI submission accession No. KP195020.1; GI:768678299)
and has published 13 research papers in national and international journals includ-
ing BMC Genomics, Plant Physiology and Biochemistry, and SpringerPlus, 3 book
chapters in Springer Nature, and 1 Patent (Patent No. 3177DEL2010), besides pre-
sentation of the researches in the national and international conferences. He is also
the guest reviewer of international journals like BMC Biotechnology, Protoplasma,
and Plant Cell, Tissue and Organ Culture (PCTOC).
xiii
xiv About the Authors
Pooja Taak is currently pursuing her PhD at Lovely Professional University (LPU),
Punjab, India, under the guidance of Dr. Bhupendra Koul, Assistant professor,
School of Bioengineering and Biosciences, Lovely Professional University, Punjab,
India. She completed her specialization in crop genetics and plant breeding during
her master’s studies in botany at Punjabi University Patiala and her project training
at the Biotechnology Division of CSIR-IHBT, Palampur, India, under the supervi-
sion of Dr. Kunal Singh. Presently, she is working on effective weed management
strategies to enhance the yield of Stevia rebaudiana. She has also published her
research findings in national and international journals.
Abbreviations
ACC deaminase 1-Aminocyclopropane-1-carboxylic acid deaminase

ACT2p Actin promoter
AM Arbuscular mycorrhizal
AOPs Advanced oxidation processes
ArsC Arsenate reductase gene
BCF Bioconcentration factor
BHC Benzene hexachloride
BIS Bureau of Indian Standards
BTEX Benzene, toluene, ethylbenzene, and xylene
CAS Community Approaches to Sanitation
CAX-2 Calcium vacuolar transporter
CEC Cation exchange capacity
CUP1 Copper metallothionein
CNTs Carbon nanotubes
DAF Dissolved air flotation
DCE 1,2-Dichloroethene
DDT Dichlorodiphenyltrichloroethane
DGEF Directorate General of Foreign Trade
DPT Dipropyltryptamine
γ-GCS γ-Glutamylcysteine synthetase gene
EDTA Ethylenediaminetetraacetic acid
ERH Electrical resistance heating
EUROPHYT European Union Notification System for Plant Health
Interceptions
EXIM Export Import Foreign Trade
FDA Food and Drug Administration
FSSAI Food and Safety Standard Authority of India
GIS Geographical information system
eSCAP Soil contamination assessment prototype
GSH Glutathione
IASWC Indian Association of Soil and Water Conservationists
xv
xvi Abbreviations
IBI International biochar initiative

ISCO In situ chemical oxidation
ISV In situ vitrification
ISPV In situ plasma vitrification
ITC-HS Indian Trade Classification-Harmonized System
Mer Mercuric reductase gene
MFCs Microbial fuel cells
MTs Metallothioneins
MHz Megahertz
NAAT Nicotianamine aminotransferase genes
NtCBP4 Nicotiana tobaccum calmodulin-binding protein
OASIS Operational and Administrative System for Import Support
PAA Poly(amidoamine)
PAH Polycyclic aromatic hydrocarbon
PCs Phytochelatins
PCBs Polychlorinated biphenyls
PCE Perchloroethylene
PCP Pentachlorophenol
PGP Plant growth promoting
PCs Phytochelatins
PMET Pittsburgh mineral and environmental technology
PPB Parts per billion
PRB Permeable reactive barrier
psMTA Pisum sativum metallothionein like
RASFF Rapid Alert System for Food and Feed
RFH Radio-frequency heating
RDX Royal demolition explosive
RS Remote sensing
SCSI Soil Conservation Society of India
SDS Sodium dodecyl sulfate
SEZ Special economic zones
SRS1p Soybean rubisco promoter
SRB Sulfate-reducing bacteria
SVE Soil vapor extraction
SVOCs Semi-volatile organic compounds
SWaCH Solid waste collection and handling
TCE Trichloroethylene
TF Translocation factor
TNT Trinitrotoluene
TEPP Tetraethyl pyrophosphate
TPH Total petroleum hydrocarbon
USEPA United states environmental protection agency
VC Vinyl chloride
VOCs Volatile organic compounds
WHO World health organization
Abbreviations xvii
YCF1 Cadmium factor protein

ZAT Zinc transporter of Arabidopsis thaliana
ZVI Zero-valent iron
nZVI Nano zero-valent iron
Chapter 1
Soil Pollution: Causes and Consequences
Abstract There has been a rapid rise in the soil pollution over the last two decades
which has posed threat to living beings and the ecosystem as well. Soil pollution is
caused by both natural and anthropogenic activities. Former includes volcanic erup-
tions, earthquakes, tsunamis etc. while the later includes metals (trace and heavy
metals), chemicals and radioactive wastes. The chemicals can be grouped into pes-
ticides and allied chemicals, crude petroleum and its derivatives and polymers, plas-
ticizers and other wastes. Radioactive wastes include nuclear power generation
wastes and other by products released from nuclear technology (medicines and
research). These are harmful substances which stay in the ecosystem for long dura-
tion during which they get accumulated and biomagnified to concentration poten-
tially toxic to organisms at higher trophic levels in the food chain. Most of these
chemicals are carcinogenic, teratogenic and mutagenic in nature. It is therefore cru-
cial to develop tools to assess potential risks of human exposure to pollutants and to
decide threshold concentrations in soils in order to protect human health.
Monitoring of soil quality is a difficult process because of the scarcity of moni-
toring variables and other indicators. The alarming situation of the state of soil pol-
lution has forced the scientific community to develop innovative, reproducible
strategies/technologies (in situ or ex situ) for treating polluted soils. This chapter
summarizes various aspects of soil pollution, its causes and consequences.
Keywords Pollution · Toxic effects · Heavy metals · PAHs
1.1 Introduction
In the coming years, sustainable plant productivity and availability of bio-resources

is expected to become a major constraint for food and nutritional security for human
population in developing countries where per capita arable land is shrinking while
human and livestock population is steadily increasing (Koul 2014; Marchand 2017).
Over-exploitation of bio-resources due to modernization and desire for comfort has
led to a state where comforts are encapsulated by pollution(s), hazards, and
© Springer Nature Singapore Pte Ltd. 2018 1

B. Koul, P. Taak, Biotechnological Strategies for Effective Remediation
of Polluted Soils, https://doi.org/10.1007/978-981-13-2420-8_1
2 1 Soil Pollution: Causes and Consequences
Fig. 1.1 Types of pollution
depletion of biodiversity. Urbanization, modern agricultural practices, industrializa-

tion, transportation, R&D work, hospital wastes, domestic wastes, mining and
radioactive wastes have led to the deterioration of our immediate environment at an
alarming rate (Gautam and Agrawal 2017; Kumar and Chopra 2017). Generally,
pollution can be classified on the basis of the environment in which it occurs and
can be grouped as air, water, noise, thermal and radioactive pollution (Fig. 1.1).
Among these pollution types, soil pollution has become a matter of global concern
1.2 Causes of Soil Pollution 3
as these factors have cumulative effect on the environment and human health
through food chain-contamination.
Land is one of the most important components of Mother Earth. Land provides a
base for the growth of vegetation and other activities. Various natural and anthropo-
genic activities like, construction of buildings, use of pesticides and chemical fertil-
izers, waste disposal, volcanic eruptions, tsunamis etc. are invariably responsible
for soil degradation. Thus, unwanted change in the physical, chemical and biologi-
cal characteristics of soil due to natural as well as anthropogenic activities is known
as soil pollution. Studies have revealed that 15% of the total arable land of the world
has been extensively degraded due to the mixing of several toxic pollutants (Wuana
and Okieimen 2011; Li et al. 2014) into the soil. Illegal dumping of the solid wastes,
polluted water absorbed by the soil, use of chemical fertilizers and pesticides,
dumping of minerals, oils, and radioactive wastes are the major causes of soil pol-
lution (Cachada et al. 2018). Soil fertility is severely affected due to the above-
mentioned factors. Till date, soil pollution has spread in many developed and
developing countries of the world.
At present, heavy metals are the most perilous soil contaminants because they are
persistent, environmentally stable and capable of producing toxic effects on human
health and its immediate environment (Rascio and Navari-Izzo 2011). Heavy metals,
though not required for normal plant growth, but are absorbed easily from the soil,
cause phytotoxicity by inactivating enzymes, which further inhibit the plant physi-
ological activities (Rascio and Navari-Izzo 2011). Expansion of industrial areas,
improper heavy metal waste disposal, uncontrolled use of animal manures, fertiliz-
ers, pesticides, sewage sludge, coal combustion residue, petrochemical spillage,
untreated effluents from refineries and tanneries are the main source for the release
of these hazardous wastes (Khan et al. 2009; Zhang et al. 2010). Soil and water turns
into a real sink for toxic metals that are discharged by various anthropogenic activi-
ties. Among the list of notorious heavy metals, metals such as zinc (Zn), chromium
(Cr), nickel (Ni), lead (Pb), cadmium (Cd), arsenic (As), copper (Cu) and mercury
(Hg) find the top position (GWRTAC 1997). Compared to the organic contaminants,
heavy metals do not undergo chemical or microbe-mediated degradation and so they
are more persistent in the soil than their organic counterparts. Therefore, heavy
metal remediation has become a tedious task (Kirpichtchikova et al. 2003; Adriano
2003). Moreover, these toxic heavy metals also hinder the degradation of organic
pollutants (Maslin and Maier 2000). The contaminated soils are a risk to all living
beings and the environment through direct contact or ingestion of contaminants and
drinking of polluted ground water (McLaughlin et al. 2000a, b; Ling et al. 2007).
1.2 Causes of Soil Pollution
As mentioned before, there are mainly two causes of soil pollution, (a) natural and
(b) anthropogenic (Fig. 1.4). The former includes volcanic eruptions, earthquakes,
tsunamis etc. while; the later includes metals (trace and heavy metals), chemicals
and radioactive wastes. The chemicals can be grouped into pesticides and allied
chemicals, crude petroleum and its derivatives and polymers, plasticizers and other
wastes. Radioactive wastes include nuclear power generation wastes and other by
product released from nuclear technology (medicines and research). These are
harmful substances which persist in the environment for long duration during which
they are taken up by the atmosphere, accumulated and biomagnified to concentra-
tion potentially toxic to organisms at higher toxic levels in the food chain.
1.2.1 Natural Sources
Natural sources which are responsible for soil pollution include earthquakes, volca-
nic eruptions, alterations in rainfall patterns and geographical changes. Natural
accumulation of chemicals (for e.g. accumulation of perchlorate in soil) and toxic
compounds in the soil surface leads to soil pollution.
1.2.1.1 Volcanic Eruptions
Eruption of hot materials such as lava, dust, gases and ash particles from volcano is
termed as volcanic eruption. According to U.S Geological survey volcano produces
large amount of hazardous substances that destroy the surrounding areas and are
toxic to living beings. Large quantity of greenhouse gases (sulphur dioxide, carbon
dioxide, hydrogen sulphide, carbon monoxide, hydrogen fluoride) and other aero-
sols are released during volcanic eruptions. These gasses accumulate in the atmo-
sphere, this phenomenon is termed as “outgassing”. Lava and ash particles released
from volcano destroy the surrounding land. Lava destroys the fertile top layer of the
soil which is termed as soil erosion through volcanoes.
1.2.1.2 Earthquakes
Earthquakes are the sudden release of energy in earth’s surface which is caused by
the movement of tectonic plates below the soil surface or by other volcanic activi-
ties. Earthquakes damage the infrastructure, pipelines, bridges, embankments,
dams, road lines, railway lines etc. furthermore, they cause severe damage to the
Earth’s surface, contaminate groundwater, causes landslides, mudflow etc.
Agricultural soil becomes non-fertile after earthquakes. Secondary effects of earth-
quakes include the damages which are caused by broken water, fuel, electrical and
oil pipelines (manmade materials). Toxic materials from sewage, medical and radio-
active wastes accumulate in the soil, water and air. Release of radioactive material
from nuclear power plant of Japan during March 2011 earthquake is an example of
the secondary effect. One major effect of earthquake is “soil liquefaction” which
occurs when soil stiffness and strength is reduced to zero and soil gets converted
into liquid or mud form. Soil liquefaction causes severe damage to residential areas
and living beings. Various reports are available on damage caused by soil liquefac-
tion, such as Niigata and Alaska earthquake in 1964, Loma Prieta earthquake in
1989, and Great Hanshin earthquake in 1995. More recently, liquefaction caused
severe damage to Christchurch townships of New Zealand in 2010.
1.2.1.3 Alterations in Rainfall Patterns
Organic materials are the most important constituent of soil as it frames soil struc-
ture, balance the nutrients, oxygen and water holding capacity of soil particles.
Alterations in rainfall pattern significantly affect the composition of organic materi-
als in the soil as organic matter is sensitive to climate alterations. Furthermore,
excessive rainfall on acidic soil increases acidification and leaching of the soil (Wild
1993). Arid and sub humid areas are greatly affected by variations in climate. For
example acid sulphate soils are formed from oxidation of pyrite rich materials due
to excessive rainfall (Rounsevell and Loveland 1992).
1.2.1.4 Geographical Changes
Changes in the geographical factors impose various adverse effects on the soil qual-
ity. The connection between soil erosion and geographical factors can be monitored
with the two recently developed techniques i.e. geographic information system
(GIS) and remote sensing techniques (RS). These techniques can be used to obtain
the information on soil quality and heterogeneity of soil surface as well as to inves-
tigate the extent of land degradation.
1.2.1.5 Tsunamis
Tsunami is a series of waves in the ocean which are caused by the displacement of
huge amount of water. Underwater explosions, earthquakes, landslides and volcanic
eruptions and other disturbances below or above the water level can cause tsunami.
Tsunami is responsible for salting of drinking water and agricultural land. Large
amount of different pollutants are carried away with the flooded water and get
deposited over the soil surface (Moqsud and Omine 2013). These deposits cause
land and water pollution.
1.2.2 Anthropogenic Sources
Various wastes including municipal, agricultural, domestic, industrial etc. are the
major anthropogenic source of soil pollution (Alloway 1995). All these wastes can-
not be considered as pollutant as they can be further recycled into some useful
materials. For e.g. animal excreta and crop residues can be recycled to manure,
papers and plastics can be converted into other products. Most commonly, these
wastes are considered as useless to human beings. Following are the major man-
made sources of soil pollution:
1.2.2.1 Solid Wastes
Solid wastes are the discarded or abandoned materials which are of no use. Fiscal
and population growth lead to huge amount of solid waste production in urban areas
(Karishnamurti and Naidu 2003; Singh et al. 2011). These wastes can be further
classified depending on their source i.e. municipal waste, hospital wastes and indus-
trial wastes.
Municipal Solid Wastes (MSW)
Municipal solid wastes comprise of domestic waste, market wastes, kitchen wastes,
slaughterhouse wastes, livestock and poultry wastes, ceramic wastes, glass and met-
als (DOE 2002). Municipal wastes can be further categorized into five types based
on the kind of pollutant.
• Recyclable materials: plastics, glass, aluminium cans, metals.
• Hazardous wastes: batteries, medicines, bulbs, fertilizer and pesticide containers,
paint, old computers, cell phones, printers etc.
• Biodegradable wastes: food and kitchen wastes
• Inert waste: demolition and construction wastes
• Non-biodegradable wastes: organic compounds and some heavy metals
There are two main methods to dispose the municipal wastes, i.e. dumps and
landfills. In these days landfills are specially designed in order to prevent the pollut-
ants from entering into soil. Synthetic sheets or clay linings can be used in order to
separate the waste materials from the soil below it. In the year 2009, 1098 landfills
were used to dispose 54.3% of municipal waste produced in U.S.A, while 11.9% of
the waste was disposed by incineration. The average production of municipal wastes
for the year 2013 is shown in Fig. 1.2.
Soil pollution due to municipal wastes is a major environmental threat in devel-
oping countries due to improper waste disposal. Open disposal of municipal wastes
poses various threats to soil as well as ground water.
Hospital Wastes
Hospital wastes are produced as a result of diagnosis, treatment and immunization

of human beings and animals. These wastes includes discarded surgical instru-
ments, discarded blood, contaminated microbial cultures, body parts (amputation),
30
25
Production percentage
20
15
10
0
Metals Rubber, Glass Wood Plastics Food Yard Paper and Other
leather scraps trimmings carbboard
and
texties
Materials
Fig. 1.2 Average production of the municipal solid-waste in the year 2013
human or animal tissues, used dressings, bandages, cotton, gloves, poisonous and
corrosive chemicals, solvents, diagnostic kits, empty bottles, genotoxic waste, radi-
active wastes (liquid used for radiotherapy) etc. According to WHO (2018) 0.5 kg
of waste per hospital per bed is generated in developed countries, while 0.2 kg of
waste per hospital per bed is generated in developing countries. Improper disposal
of these wastes cause several health hazards. These wastes are most commonly
dumped in landfills with the other municipal wastes and contaminate the soil of the
sorrounding area. Incineration of hospital wastes is the alternative strategy which is
followed in developing countries. Release of pollutants during incineration of these
wastes also imposes adverse effects on public health as well as the enviornment
(Batterman 2004). Production of different organic pollutants such as PCBs, PAHs
(Fig. 1.18) dioxins and furan (Fig. 1.17) and other carcinogens as a result of incin-
eration technique leads to several health issues (Beyersmann 2002; WHO 2011).
Bottom ash which is produced after incineration consists of organic compounds and
heavy metals (Cd, Pb, Cr, Hg, Zn etc). Improper disposal of this ash affects the soil
(alter the soil chemistry) of the dumping area and pollute the environment. To over-
come various environmental related concerns of hospital wastes, there is a need to
implement some effective and environmentally sound hospital waste management
strategies for proper disposal and to reduce the environmental and public health
issues (Gidarakos et al. 2009).
Industrial Wastes
Industrial wastes comprise of concrete, scrap, paints, metals, oil, trash, chemicals
(organic and Inorganic), gravel, masonry etc. These types of wastes are the chief
soil pollutants as they are toxic, reactive or corrosive. Disposal of these wastes in an
improper way can pose serious health issues and environment related problems. In
U.S.A, drastic increase of the industrial wastes i.e. 4.5 million tons has been reported
after the second World War. In 1990, the amount of wastes reached upto 265 million
tons. It has been reported in 1980 that more than 70,000 chemicals were manufac-
tured by different industries in U.S.A alone. High toxicity of these chemicals affect
the chemical and physical nature of the soils and their fertility. Toxic contaminants
settle down in the soil and are accumulated by the crops growing on it. Furthermore,
with an increase in the population size, urbanization and construction activities are
on the rise. These activities are the major cause of deforestation which are further
responsible for soil erosion and ultimately cause soil pollution. The developed
countries have devised strategies for remediation of industrial wastes while, the
developing countries are still not able to invest in pollution control methods
(Fig. 1.3).
1.2.2.2 Agricultural Practices
In the modern era, human population is increasing while the amount of arable land
to feed the teeming millions is decreasing. In order to cope with increasing demand
of food, various types of agrochemicals are used to protect the crops (from patho-
gens and insects) and to enhance the crop yield. Agrochemicals can be classified as
pesticides (herbicides, insecticides and fungicides), fertilizers, hormones and
Causes of soil pollution
Natural Anthropogenic
Volcanic eruptions Solid wastes
Agricultural practices
Earthquakes
Chemical wastes
Geographical changes
Radioactive wastes
Changes inrainfall pattern
Mining and smelting
Tsunamis Construction activities
Fig. 1.3 Causes of soil pollution

Organophosphorus
Organochlorine
Carbamates
Pyrethroid
Insecticides
Organochlorine
Pesticides Herbicides
Agrochemicals Aniline derivatives

Fungicides
Fertilizers
Fig. 1.4 Agrochemicals responsible for soil and water pollution
animal manure. These chemicals are highly toxic at a dose more than threshold
value and pose serious environmental and health risks. Application of manures and
fertilizers introduce heavy metals and other organic pollutants into the soil, which
alter its composition and retards its fertility. Ar, Mn, Cd, U, Vn, Zn are the heavy
metals which are associated with phosphate fertilizers and animal (poultry and pig)
manure. The agrochemicals which are used to eliminate or repel the pests are called
pesticides. Pesticides can be natural, synthetic or modified chemicals or other organ-
isms that can work as biological control agent. According to British Food and
Environment Protection Act, 1985, pesticide can be defined as a substance used to
destroy undesirable plants, and the organisms which are harmful to plants or plant
products. Pesticides are broadly divided into three categories i.e. herbicides, insec-
ticides and fungicides. The consumption pattern of pesticides is shown in Fig. 1.5.
Pesticides can persist for longer duration in soil and water. Various pesticides are
recalcitrant in nature and are non-biodegradable (Navarro et al. 2007).
Herbicides
Herbicides are the chemicals which are used to inhibit the growth of undesirable
plants in a field of cultivated crop. Persistence of herbicides in soil depends upon the
soil characteristics such as soil pH, soil nutrient composition etc. Application of
heavy doses on soil can cause a decline in soil flora and fauna. More than fifty
chemical groups of organic and inorganic compounds are used as herbicides.
Herbicides can be categorized on the basis of their chemical structures such as:
• Organochlorine herbicides (Fig. 1.6): These are the derivatives of phenoxyacetic
acid for e.g. 2,4,5-trichlorophenoxyacetic acid, 2,4-dichlorophenoxyacetic,
2-methyl-4,6-dichlorophenoxyacetic acid. These herbicides also contain the

derivatives of aniline such as alachlor and propanil (Fig. 1.7).
• Organophosphorus herbicides: These are the most widely used herbicides in
agriculture and are also known as glyphosates. They are non-carcinogen and
have high weed killing efficiency. Most commonly used herbicide “glyphosate”
(Fig. 1.8b) is a modified form of glycine (Fig. 1.8a). Certain derivatives of urea
(Fig. 1.9), triazine (Fig. 1.12) and carbamic acid are also used as herbicides.
0.5 %
21 %
Insecticides
44 %
Herbicides
Fungicides
30 %
Other
Fig. 1.5 Consumption pattern of pesticides in the world
(A)
(B) (C)
Fig. 1.6 Structures of organochlorine herbicides: (a) Phenoxy acetic acid, (b) 2,4 dichloro
Phenoxy acetic acid and (c) 2,4,5 trichloro Phenoxy acetic acid
(A) (B)
Fig. 1.7 Structures of aniline derivatives: (a) Propanil and (b) Alachlor
Fig. 1.8 Structures of (a)

Glycine and (b)
Glyphosate
(A) (B)
(A) (B) (C) (D)
Fig. 1.9 Structures of urea derivatives: (a) Diouron, (b) Fluometuron (c), Linuron and (d)
Chlorobromuron
Insecticides
These are the chemicals which are used to kill the insects in crop fields. Insecticides
include larvicides and ovicides which kill insect larvae and eggs respectively. Use
of various insecticides in agriculture has been rapidly increasing over the past two
decades. Insecticides can be divided into four main groups on the basis of the
organic compound present.
• Organophosphorus compounds (Fig. 1.15): These are the nerve poisons and
effect on central nervous system. They can be applied as fumigants, systematic
pesticides or contact poisons in agriculture. However, these are highly toxic to
non-target organisms also. Sarin, tetraethyl pyrophosphate (TEPP), diazinon,
dibrom, malathion, temephos, chlorpyrifos and terbufos are the most commonly
used organophosphates.
• Organochlorines (Fig. 1.16): These are easily available and highly effective
insecticides to control the pests which are responsible for yellow fever and
malaria. They can be classified into three different classes i.e. BHC, DDT and
cyclodiene. Organochlorines have been reported as persistent organic com-
pounds which kill non-target organisms also.
• Carbamates (Fig. 1.10): These organic compounds are the derivatives of car-
bamic acid (NH2COOH) and are also known as urethanes. Terbucarb, propoxur,
carbaryl, aldicarb, oxamyl and carbamates are some commonly used agricultural
insecticides. However, they have been proved to be fatal for various bird species
and kill the non-target insect species by inactivation of acetylcholinesterase
enzyme.
• Pyrethroids: Pyrethroids consists of six active constituents i.e. jasmolines I and
II, pyrethrum I and II and cinerins I and II (Fig. 1.11). They have lower persis-
O
H H
H
O C N
O C N C N
CH3
CH3 CH3
O
O
N CH3 O
HC C S CH3
CH3
(A) (B) (C)
Fig. 1.10 Structures of Carbamate pesticides: (a) Aldicarb, (b) Carbofuran and (c) Carboryl
(A) (B)
(C) (D)
(E) (F)
Fig. 1.11 Structures of natural Pyrethrins: (a) Cinerin I, (b) Cinerin II, (c) Jasmolin I, (d) Jasmolin
II, (e) Pyrethrin I and (f) Pyrethrin II
tence and toxicity, while higher insecticidal activity. Bioresmethrin, fenvalerate,

allethrin and permethrin are the four groups of synthetic pyrethroids. These
degrade easily in the sunlight and at high temperatures. However, they are very
toxic to bees and aquatic flora and fauna.
Fungicides
These are the natural or synthetic, organic or inorganic chemical compounds which
are used to prevent fungal growth. They are used as fumigants. Bordeaux mixtures,
organotins (Fig. 1.13), phthalimide (Captan) (Fig. 1.14a), tributyltinacetate and
benzimidazole (Fig. 1.14b) are some most commonly used agricultural fungicides.
(A) (B) (C)
Fig. 1.12 Structures of triazine derivatives: (a) Triazine, (b) Atrazine and (c) Simazine
(A) (B)
Fig. 1.13 Structures of organotin fungicides: (a) Tributyltinacetate and (b) Triphenyltinacetate
(A) (B)
Fig. 1.14 Structures of (a) Captan and (b) Benzimidazole

Fig. 1.15 Structures of
organophosphorus
pesticides (a) TEPP (b)
Sarin
(A) (B)
(A) (B) (C)
(D) (E) (F)
Fig. 1.16 Structures of organochlorine pesticides: (a) DDT, (b) BHC, (c) Chlorinated cyclodiene,
(d) Aldrin, (e) Dieldrin and (f) Heptachlor
(A) (B)
(C) (D)
Fig. 1.17 Structures of dioxins and furans: (a) 2,3,7,8-tetrachlorodibenzo-p-dioxin, (b) 2,3,7,8-tetra-
chlorodibenzofuran, (c) 1,2,3,7,8-pentachlorodibenzo-p-dioxin and (d) 2,3,7,8-tetrachlorodibenzofuran
(A) (B) (C)
(D) (E) (F)
(G) (H) (I)
Fig. 1.18 Structures of PAHs: (a) Pyrene, (b) Benzo(c) Phenanthrene, (c) Triphenylene, (d)
Phenanthrene, (e) Benz(a) Anthracene, (f) Chrysene, (g) Naphthalene, (h) Anthracene and (i)
Tetracene
1.2.2.3 Radioactive Wastes
These wastes contain radioactive materials which are the byproducts of nuclear
power plants and research stations. Although, the development of nuclear technol-
ogy is one of the greatest achievements of the last century but, the wastes generated
by it is severely lethal to all living beings and are generally monitored by govern-
mental agencies so as to protect the environment and public health. Radionuclides
are the unstable elements having atomic number > Bi-83 (Bismuth) and are radioac-
tive (Igwe et al. 2005). These elements release energy in the form of ionizing radia-
tions (van der Perk 2006). Cesium-137, iodine-131, americium-241, cobalt-60,
strontium-90, plutonium, radontechnetium-99, tritium, radium, uranium and tho-
rium are some naturally occurring radionuclides. Emissions of radionuclides from
nuclear power stations are the sources of anthropogenic nuclear wastes. Cesium-137
and strontium-90 have half-life of 30 and 29.1 years respectively. It has been
reported that the average levels of radionuclides in the soil was 620 mc km−2 for Cs
137 and 388 mc km−2 for Sr 90 (Igwe et al. 2005). Deposition of these radionuclides
in the soil will lead to the magnification of radioactive materials through food chain
contamination and ultimately affecting the living beings. Soil contaminated with
radioactive materials, affect its chemical and physical characteristics and also its
fertility. Polluted soils lose their fertility due to the toxic effects of radioactive mate-
rials. The mobilization of radionuclides in the soil depends upon its pH, composi-
tion, and rainfall. Radioactive materials are easily taken up by the plants and get
accumulated into the living beings through the ingestion of the contaminated plant
products.
1.2.2.4 Chemical Wastes
During the past decade, there has been a drastic increase in the amount of chemical
wastes generated through industrialization in the developed as well as the develop-
ing countries. Chemical wastes include polynuclear aromatic hydrocarbons, petro-
leum hydrocarbons, various solvents and other heavy metals. It can be classified
into organic and inorganic chemical wastes. Organic chemical wastes include
hydrocarbons (aliphatic and aromatic) and inorganic chemical wastes include heavy
metals. Organic wastes impose several health risks such as, toxicity to immune,
reproductive and nervous system, cause allergies and cancers (van der Perk 2006).
Some chlorinated aromatic compounds for e.g. pentachlorophenol, hexachloroben-
zene, polychlorinated biphenyls and some VOCs such as dichloroethylene, trichlo-
roethylene, tetrachloroethylene and vinyl chloride are carcinogenic in nature.
Organic pollutants can transform in soil through various methods such as volatiliza-
tion, leaching or chemical, physically or biological transformation (Semple et al.
2003). Furthermore, organic pollutants undergo bioaccumulation in living organ-
isms. The accumulation of contaminants from soil to living beings is termed as
bioaccumulation. Accumulation of organic contaminants in earthworms is linked to
bio magnification as these contaminants enter the food chains and cause various
adverse effects at each trophic level.
Heavy metals are the class of elements which having metallic properties. It
includes metalloids, lanthanides and actinides. These metals are characterized by
atomic number < 2, density range 3.5–7 g/cm3 and atomic weight 22.98 to <40 (Afal
and Wiener 2014). In the past decade heavy metals have considerably affected the
soil productivity and quality with an increase in pollution from agricultural, indus-
trial and municipal sources. Soils polluted with heavy metals poses various adverse
effects on living beings and soil fertility (Smith et al. 1995). These metals possess
specific thermal and electrical conductivities. In nature, there are 92 elements, out
of which 30 are the metalloids. Among these, chromium (Cr), lead (Pd), molybde-
num (Mo), beryllium (Be), boron(B), lithium (Li), tellurium (Te), antimony (Sb),
aluminium (Al), titanium (Ti), vanadium (V), manganese (Mn), tungsten (W),
cesium (Cs), cobalt (Co), nickel (Ni), copper (Cu), selenium (Se), arsenic (As),
strontium (Sr), silver (Ag), cadmium (Cd), tin (Sn), barium (Ba), platinum (Pt), gold
(Au), mercury (Hg) and bismuth (Bi) are toxic for human beings, at higher concen-
1.3 Harmful Effects of Soil Pollution 17
tration (Mingho 2005). Heavy metals such as Pb, Cd, Cr, Cu, Ni, and Zn etc. can
change the soil chemistry and impose toxic effects (Table 1.1) on living beings
(Shaylor et al. 2009).
1.2.2.5 Mining and Smelting
Mining and smelting wastes comprise the wastes produced during metal and min-
eral extraction and their processing. These wastes damage the surrounding environ-
ment and harm all living beings, as air borne pollutants. Various researches have
been conducted regarding the management of mining and metallurgical sites so as
to neutralize the toxic effects of the pollutants on the environment. Toxic metals
which are found in tailings of mining and smelting industries include Hg, Pb, Ar,
Cu, Cd, Se, Ni, Zn and Cn (Ryszka and Turnau 2007; Escarré et al. 2011; Asensio
et al. 2013; Kapusta and Sobczyk 2015). Wastes from mining and smelting indus-
tries include brown and red mud from bauxite refining, phosphogypsum from phos-
phoric acid production plant, fluorogypsum from hydrofluoric acid production,
slags from lead, copper, and phosphorus processing industries, slags from carbon
and steel production etc. These wastes can be classified as (1) drilling muds and
wastes containing oil, (2) tailings from sulfide ore processing and (3) wastes gener-
ated from physical and chemical processing of metalliferous and non-metalliferous
minerals. 90% of these wastes are generated by fertilizer, chemical and petroleum
refining industries while the rest 10% is generated by other small-scale industries.
Various studies have shown that these wastes can stay in soil for a long duration i.e.
from hundreds to thousands of years. As a result, these contaminants gain access
into the food chain and impose several health hazards.
1.3 Harmful Effects of Soil Pollution
I. Impact on plants
Our biosphere has three zones lithosphere, hydrosphere and atmosphere. A slight
disturbance in any of the zones may affect the other zones severely because of the
interlinking network of several ecological functions operating within. Many anthro-
pogenic activities that have led to soil pollution have also affected the functional
quality of living organisms including the elimination of a species, in severe cases.
Soil pollution greatly affects the agroecosystems (Table 1.1) which can be analyzed
as follows:
(i) Loss of crop productivity and crop diversity
Pollution of soil and water are a hindrance for crop productivity and diversity
which has decreased drastically within a span of 20 years (Nath et al. 2005). The
Table 1.1 Source and toxicity symptoms of some important heavy metals
18
Oxidation
Element state Source Toxicity Reference(s)
Arsenic (As) As (III) Igneous and sedimentary On human: Carcinogenic, affects nervous, integumentary, Marin et al. (1993),
and As rocks, pesticides, herbicides, cardiovascular, immune, endocrine, renal, hepatic, hematopoietic and Barrachina et al.
(II) and insecticides reproductive system. (1995), Cox et al.
manufacturing industries On plants: Reduced leaf area, fruit yield, leaf fresh weight, seedling (1996), Abedin et al.
height and seed germination. Chlorosis, stunted growth, wilting. (2002), and Abdul
et al. 2015
Chromium (Cr) Cr(III), Electroplating processes, On human: Dermatological problems, nephrotoxic, teratogenic Sharma and Sharma
Cr(IV), stainless steel industries, effect, digestion problems, upper respiratory and pulmonary tract (1993), Moral et al.
Cr3+ and leather tanning, mordant disorders, induce mutations. (1995), Panda and
CrO42− dyeing, wood preservation, On plants: Reduced bud sprouting, germination percentage, root Patra (2000),
ore refining length, plant height, leaf number, leaf area, yield reduction, grain Nematshahi et al.
weight reduction, and increased seed deformity. (2012), and Langård
and Costa (2015)
Nickel (Ni) Ni2+, Ni3+ German silver, stainless steel, On human: Acute toxic, chronic and sub chronic toxic, genotoxic, Khalid and Tinsley
1
electroplating and ceramic developmental toxic, haematotoxic, immunotoxic, neurotoxic, (1980), Sheoran et al.
industries, reproductive toxic, carcinogenic. (1990), Pandolfini
On plants: Germination reduction, photosynthesis, metabolism and et al. (1992),
cell structure disruption, oxidative stress. Barsukova and
Gamzikova (1999),
Lin and Kao (2005),
and Klein and Costa
(2015)
Lead (Pb) Pb Fossil fuel combustion, On human: Attack brain and central nervous system which leads to Moustakas et al.
(NO3)2, automobiles exhaust coma, convulsions and even death, mental retardation and behavioral (1994), Kabir et al.
Pb(II) disorders, hypertension, anemia, renal disorders, immunotoxic, (2009), Hussain et al.
reproductive toxic. Effects on the mental development of infants. (2013), Skerfving and
On plants: Growth retardation, chlorosis, blackening of roots, Bergdahl (2015)
disturbs water balance and mineral nutrition, retard photosynthesis,
effect hormonal balance, membrane structure and permeability.

Oxidation
Manganese(Mn) Mn(II) Ceramics, electrical coils, On human: Effects respiratory tract and brain, symptoms include Shenker et al. (2004),
batteries, welding rods, forgetfulness, hallucinations, dullness, weak muscles, insomnia, Asrar et al. (2005),
matches, glasses, dyes headaches and nerve damage, also cause lung disorders and Doncheva et al.
Parkinson disease. (2005), Arya and Roy
On plants: Disturbance in water balance, swelling of cell walls, leaf (2011), and Lucchini
wilting, brown spots on leaves. et al. (2015)
Silver (Ag) Ag(II) Electrical appliances, plating On human: Corneal injury, skin irritation, dermatitis, breathing Holler et al. (2015),
applications, silver brazing, disorders, dizziness, respiratory irritation, headaches, stomach Wang et al. (2015),
manufacturing of mirrors and disorders, confusions, coma and even death. Cox et al. (2016), and
batteries On plants: Reduced chlorophyll content and biomass of plants, Zuverza-Mena et al.
disturb the metabolism of plants. (2016)
Zinc (Zn) Zn(II) Mining and smelting, sewage On humans: Oral intake can cause nausea, pain, vomiting cramps Bonnet et al. (2000),
effluents, municipal wastes, and diarrhea. “zinc shakes” or “zinc chills” or metal fume fever can Doncheva et al.
wastes from automobiles, occur by inhaling the freshly formed zinc oxides. (2001),
batteries, pesticides, steel and On plants: Chlorosis and reddening of young leaves, brown spot on Manivasagaperumal
brass alloys industries and the leaf surface, in severe cases necrotic lesions appears on the leaves et al. (2011), and
construction sites and leaf death occurs, stunted growth of main root and lateral roots Sandstead (2015)
become yellow in color.
Cadmium (Cd) Cd2+ Mining and smelting, sewage On human: Bone demineralization, liver Disturbance, lung cancer, Jiang et al. (2001),
sludges, batteries, plating, renal tubular dysfunction, Itai-Itai disease. Wang et al. (2007),
pigments and plastic On plants: Reduction in seed germination, plant nutrient content, Bernard (2008),
industries shoot and root length. Ahmad et al. (2012),
Kang et al. (2013),
and Yourtchi and
Bayat (2013)
(continued)
19
Table 1.1 (continued)
20
Oxidation
Copper (Cu) Cu2+, Dyes, paints, pigments, On humans: Vomiting, hematemesis (blood vomiting), hypotension Kjær and Elmegaard
Cu3+ ceramics and pesticide (low blood pressure), jaundice, gastrointestinal disorders, damage to (1996), Cook et al.
manufacturing industries kidney and liver. (1997), Sheldon and
On plants: Interveinal foliar, cream or white spots or lesions on Menzies (2005), and
leaves, in severe cases wilting can occur. Purpling of foliage. Ellingsen et al. (2015)
Iron (Fe) Fe2+, Fe3+ Mining and smelting, steel On humans: Damage gastrointestinal tract, stomach ache, nausea Becker and Asch
industries and blood vomiting, metabolic acidosis damage brain and liver, (2005), Nagajyoti
hypovolemic shock, death may occur due to liver failure. et al. (2010), and
On plants: Bronzing and stippling of leaves, leaf discoloration. Ponka et al. (2015)
Mercury (Hg) Hg1+, Fossil fuel combustion, On humans: Affect gastrointestinal, neurologic and renal organ Du et al. (2005),
Hg2+ plastics and electronic system, sensation disturbance and lack of coordination, formication, Kibra (2008), Shekar
industries skin discoloration, swelling and desquamation, hypotonia, kidney et al. (2011), and
disorders and insomnia. Berlin et al. (2015)
1
On plants: Abnormal germination and hypertrophy of the roots and

coleoptiles, affects light and dark reaction of photosynthesis.
Cobalt (Co) Co2+, Alloys, paints, dyes and On humans: Cardiomyopathy, deafness, nerve problems, tinnitus, Jayakumar et al.
Co3+ pigments manufacturing thyroid and blood thickening. (2007, 2008),
industries On plants: Affect growth and plant metabolism, inhibit differentiation Jayakumar et al.
of starch grain and alter the number and structure of chloroplast per (2013), Catalani et al.
unit area of leaf, leaf fall, discoloration, reduced shoot weight. (2012), and Lison
(2015)
Selenium (Se) Se−2, Fossil fuel combustion, alloys On human: Nausea, diarrhea, tiredness and irritability, paresthesia, Alexander (2015) and
Se+6, Se+4 and rubber manufacturing hyperreflexia, discoloration and loss of teeth, nail brittleness, Gupta and Gupta
industries deformation and loss of nails, alopecia, skin rashes, joint pain. Acute (2017)
toxicity causes myocardial infarction, renal failure, vascular
symptoms such as tachycardia and flushing of face, neurological
features including tremors, myalgia and irritability.
On plants: Chlorosis, leaf become white, stunted growth of plant,
decreased protein synthesis, withering and drying of leaves and

premature death of plant.
Oxidation
Bismuth (Bi) Bi+3 Metallurgical, thermal and On humans: Affects kidney, liver and bladder, respiratory and skin Fowler et al. (2015)
electrical processes, disorders.
cosmetics and On plants: Non-toxic.
pharmaceuticals industries
Antimony (Sb) Sb(III) Alloys, plastics, rubbers, On human: Respiratory irritation, pneumoconiosis, spots on the Paoli et al. (2013) and
and textiles, paper, glass, ceramic skin, gastrointestinal symptoms, chronic bronchitis, chronic Tylenda et al. (2015)
Sb(V) and paints industries, lead emphysema, tuberculosis, pleural adhesions, cardiovascular and skin
storage batteries, related disorders, genotoxic and carcinogenic.
On plants: Decrease in leaf and root biomass production.
21
yields and the shelf-life of the traditional crops such as soybean, gram, wheat and
garlic grown in polluted areas, get affected.
(ii) Decline in nutrient use efficiency (NUE)
A low nutrient use efficiency is an indicator of more fertilizer requirement for
optimal productivity. The contaminants may interact with the soil components, soil
biota and the plant roots in several ways to affect the nutrient transformation and
absorption by the roots.
(iii) Quality of agricultural output (vegetables, fruits and grains)
The shelf-life, taste and aroma of crops grown on polluted soils gets severely
affected. Certain vegetables have shown lower storage life and give foul smell dur-
ing cooking and unpleasant taste after cooking. The oxidative damage (lipid peroxi-
dation and H2O2 levels) to the vegetables was observed when irrigated with industrial
waste waters. Interestingly, the accumulation of heavy metals in the edible parts of
the vegetables was more as compared to the grains (Hassanein et al. 2013). Not only
the vegetables and grains, the fruits of the trees that were irrigated with polluted
waters emit foul smell during short-term storage.
(iv) Heavy metal contamination of plants and plant-based products
Heavy metals in agricultural soils are derived from mining, smelting, vehicle
exhaust, as well as applications of pesticides and fertilizers (Vieira da Silva et al.
2016; Jiang et al. 2017). It has been reported that the plants grown in contaminated
and waste water irrigated land uptake and accumulate heavy metals to a level which
is toxic for consumption (Sachan et al. 2007; Chary et al. 2008).
(v) Rejection of export consignment
In continuation to the aforementioned effect, soil pollution can lead to an adverse
impact on a country’s export of food items. The food export consignments may be
rejected due to the presence of pesticides, heavy metals etc. (see Boxes 1.1 and 1.2).
(vi) Soil compaction
Soil friability and soil organic matter are important parameters which influences
root and plant growth and thus enhances the crop yield. On the other hand, soil
compaction (due to high mineral content and lower organic matter) affects the root
proliferation, penetration and nutrient and water uptake which further leads to crop
yield penalty.
(vii) Decline in available soil moisture content
The maintenance of soil moisture content is crucial for optimal yield and fertil-
izer use efficiency. Agricultural soils contaminated with pollutants negatively affects
the soil structure and the soil moisture content. An increase in soil salinity also
reduces the soil moisture content. Therefore, presence of the industrial water con-
taining high salt content reduces the agricultural soil moisture content.
(viii) Poor percolation leading to water logging
Box 1.1: Quality Control of Export and Import Consignment

The EU, US, Australia, Japan and India have developed a transparent system
for maintaining the quality standards of the import and export of plants, seeds,
grains, fruits, dry-fruits, oils, spices and other unprocessed and processed
products. On receiving each consignment lab tests rae conducted for the pres-
ence of weeds, pathogens (bacteria, virus, fungi), pesticides, chemicals etc.
After lab tests the reports are displayed onto the respective web portals
wherein a detailed report with reasons for destruction, interception, withhold-
ing or returning of the consignment from exporting countries is mentioned
along with the similar incidences that have occurred in the past.
In the EU the information on the safety of the consignmets is made visible
on two web portals (a) Rapid Alert System for Food and Feed (RASFF) and
(b) European Union Notification System for Plant Health Interceptions
(EUROPHYT). The RASFF is an informative tool to disseminate the infor-
mation on serious risks that have been detected in relation to food or feed.
This enables the EU member states to exchange views and act rapidly in co-
ordination. On the other hand, EUROPHYT is a notification and rapid alert
system which deals with the interception of the consignments, of plants and
plant products such as fruits and vegetables, during inter- and intra import
within the EU.
In the case of the US, an Import Refusal Report is generated by FDA’s
Operational and Administrative System for Import Support (OASIS) and
released by the Food and Drug Administration (FDA). The reports contain the
detailed information as to why the consignment was refused and did not get
entry into the US. In Australia, the Department of Agriculture and Water
Resources releases a monthly report called the ‘failing food report’ that lists
products (country-wise) that pose a high or medium risk to public health. In
Japan, the same information (monthly statement) is released by the Ministry
of Health, Labour and Welfare, on cases of violation of their Food Sanitation
Law. This monthly report provides the details of the exporting country and the
Article of the law that has been violated.
In India, the import and export of goods is governed by Export Import
(EXIM) Policy. India’s Directorate General of Foreign Trade (DGFT) is the
chief governing body responsible for all matters related to EXIM Policy. The
Importers that get registered with DGFT obtain an Importer Exporter Code
Number (IEC). After an IEC has been obtained, the source of items for import
must be identified and declared. The Indian Trade Classification – Harmonized
System (ITC-HS) facilitates free import of most goods without an import
license. The Bureau of Indian Standards (BIS) is a committee involved with
product certification, quality system certifications and testing, and consumer
affairs. The Food Safety and Standard Authority of India (FSSAI) issues food
import clearance certificates for all food articles.
Box 1.2: Rejection of Export Consignment: Indian Case Study

There are true examples of soil pollution leading to the crop and crop product
contamination which led to the rejection of export consignments. In the recent
past, a number of Indian consignments to the US and the EU have faced noti-
fications, rejections and alerts. The main reason for rejection of plant-based
consignments is the presence of higher than approved level of pesticide resi-
due. According to the US-FDA Import Refusal Report basmati rice faced 444
refusals, plain (white or polished) rice 112, capsicum and okra 41, between
2014 and 2017, due to a higher pesticide content. Recently in 2016, Indian
cashew export consignment to the US was rejected because the laboratory test
revealed that they were contaminated by chlorphenol. The reason being that
chlorinated water was used by processing factories. The toxicity of chloro-
phenol irritates the eyes, respiratory tract and repeated administration results
in liver and bone marrow damage.
In the EU, between Jnuaru 2000 and April 2016, the RASFF portal high-
lighted 36 notifications against basmati rice, for higher than permissible con-
tent of pesticide residue such as acephate and carbendazim. From January
2014 till May 2017, 33 alerts were visible on the portal regarding the basmati
consignment which were rejected at EU ports for the presence of chemicals
such as oxytetracycline and sulphites above the permissible maximum reside
limits (MRLs). Traces of prohibited chemicals such as nitrofuran and chlor-
amphenicol were also reported. In the year 2010, the Indian consignment of
table grapes faced rejection at the EU due to the presence of chlormequat
chloride (ccc) higher than permissible level.
In Japan, the Indian shrimps export consignment were rejected due to the
presence of ethoxyquin and furazolidone, more than the acceptable MRLs.
This resulted in heavy losses to the Indian exporters.
Table 1 Notifications raised by the EU in RASFF Portal (2005–2017)

Border rejections
Serious Destructions of Border in proportion to
Country Notifications cases consignment rejections notifications (%)
China 3374 577 391 1730 51.3
Turkey 3296 874 431 2018 61.2
India 2240 672 583 1490 66.5
Brazil 1139 419 None 717 62.9
Vietnam 1049 256 136 372 35.5
In the year 2015, Indian green chillies export faced temporary rejection in
Saudi Arabia due to the presence of higher than permissible levels of pesticide
residues. In the year 2016, several concerns were raised in the UAE, on Indian
mangoes, chillies and cucumbers which had pesticide residues beyond the
(continued)
Box 1.2: (continued)

Codex Alimentarius Commission’s limits. Therefore, efforts need to made at
the grass-root level. The prohibited/hazardous chemicals and their reckless
use must not be practiced anymore inorder to gain higher yields. Moreover,
the issues related to the presence of excessive pesticide residues must be
addressed/resolved by the exporting countries themselves so as to avoid rejec-
tion of the consignments thereby making the export procedures cumbersome
and time-consuming.
Although shallow water table enhances the changes of contamination of the

ground waters but, poor infiltration/percolation leads to water logging or swampy
lands. The poor-quality water along with silica and clay of hard pans which further
prevents water infiltration.
(ix) Effect on soil fertility
Fertility of soil depends on the interaction among its physical, chemical and
biological components. Presence of pollutants in the agroecosystems greatly affects
the spoil properties (soil pH, electrical conductivity, sodium adsorption ratio, bulk
density, availability of macro and micronutrients and soil organic matter). Resulting
in the loss of soil fertility and crop productivity.
II. Impact on humans and animals
Nearly 80% of the diseases particularly in developing world can be linked with
soil and water. The soil gets highly polluted by several pathogenic organisms and
hazardous industrial effluents. Undoubtedly, soil pollution is the result of urban-
technological revolution and speedy exploitation of natural resources. There are
three common ways that humans are exposed to soil materials: (i) ingestion, (ii)
respiration and (iii) skin absorption or penetration (Brevik 2013). The harmful
effects of soil pollution on human health (Table 1.1) can be discussed under the fol-
lowing headings:
(i) Effects of industrial pollutants
Industrial wastes consist of a variety of chemicals which are extremely toxic to
both animals and humans. Different industries (tanning, dyeing, chemical, pesti-
cides industries etc.) release harmful pollutants which enter the food chain causing
a number of undesirable effects. Industrial effluents when discharged through sew-
age system poison the biological purification capacity of sewage treatment resulting
in several soil and water-borne diseases. Heavy metals present in polluted soils can
release to other ecosystems, such as groundwater, rivers, atmosphere and crops, and
consequently are hazardous to human beings and ecosystems. The heavy metals
such as Zn, Hg, Cd, etc. destroy the beneficial microorganisms present in the soil
thus, affecting its fertility. Some of the trade wastes contain pathogenic bacteria,
e.g., anthrax bacilli bacteria from tanning industry waste.
(ii) Effects of urban waste products
Not only the industrial wastes but also the urban wastes spread several chronic
diseases. About 90% of the pollution load in river system is due to faecal matter. The
wastes including household wastes, building materials, sludge, dead animal skele-
tons etc. provides excellent medium for the growth of pathogenic bacteria, viruses,
etc. Vibrio cholerae found in sewage causes cholera. Solid wastes dumping areas are
a source of revolting miasma. Suspended matter in sewage covers the soil and thus,
interferes with the soil moisture. In urban areas, anthropogenic sources of soil heavy
metals include traffic emissions (vehicle exhaust, tire wear, brake lining wear, etc.),
industrial discharges (power plant, chemical plant, coal combustion, metallurgical
industry, etc.), and municipal wastes (Wei and Yang 2010; Du et al. 2015). The use
of polluted ground water containing human excreta, sewage sludge decreases soil
fertility by killing microorganisms. The fungus, Coccidioides spp. grow and repro-
duce in and above the soil. It can grow under extreme environmental conditions,
including high temperatures, high salinity or alkalinity which most other microor-
ganisms cannot tolerate. The fungus usually enters humans through the respiratory
route by inhalation of microscopic fungal spores and causes coccidioidomycosis
(Stockamp and Thompson 2016).
(iii) Effects of radioactive pollutants
The problem of radioactive wastes dumped into the soil is more complicated as
the radioactive elements can remain active in soil for longer durations (long half-
lives). Radioactive pollutants may cause detrimental effects when food containing
radionuclides is consumed by man. The pollutants may concentrate in specific body
organs and cause undesirable effects such as vomiting, nausea, restlessness, bilious-
ness, cramps and even death. The radio nuclides such as C-14, Fe-55, Mn-54,
Co-57, etc., get concentrated in the biological systems. Radiations also affects the
soil microflora and fauna and thus leads to loss of soil fertility. The radiations also
kill the plant species. Variations in radio-sensitivity among trees and shrubs are due
to differences in their chromosome number and size.
(iv) Effects of modern agro-technology
Potassium fertilizers in soil decrease the production of a valuable nutrient ascor-
bic acid (vitamin C) and carotenes in vegetables and fruits. Reckless use of nitrog-
enous fertilizers in the fields leads to accumulation of nitrates in the soil. This can
cause diarrhea and cyanosis in children who play outdoors (Henry and Cring 2013).
Use of nitrate causes eutrophication of water bodies. But, Phosphate fertilizer are
more to blamed for this and they are considered detrimental (phosphatic fertilizers
like DAP) to crop production. Cereal crops like jawar, maize, etc., grown on alka-
line soil absorb higher amounts of fluorides and are responsible for the spread of
fluorosis. Pesticides retained in soil concentrate in crops, vegetables, etc., which
taint them to such an extent that they lose their economic value. Pesticides like
1.4 Remedies to Cope with Soil Pollution 27
DDT, endrin, etc., are known to percolate slowly through soil into ground water and
contaminate the potable water supplies. Organophosphate pesticides cause extreme
muscular weakness, tremors, dizziness and interfere in the normal metabolic activi-
ties in poisoned animals.
1.4 Remedies to Cope with Soil Pollution
The impact assessment is a crucial step required for initiating and implementing
appropriate remedial measures for restoration and sustenance of polluted soils
(Cachada et al. 2018). Soil remediation strategies aim towards various in situ and ex
situ treatment regimes that deploy physical and chemical means or natural/geneti-
cally modified microbes, plants, and animals for degradation of toxic compounds or
hazardous waste into simpler non-toxic products (Pavel and Gavrilescu 2008;
Gomes et al. 2013). Pesticides and fertilisers are major contributors to soil contami-
nation so a restriction on their usage can solve the issue of soil pollution to a certain
extent. Wherever possible, biodegradable products (such as cartons for packaging)
must be used. Items that can be re-used should not be disposed of; objects made of
paper, glass, aluminium and the like should be recycled. The major objective of soil
remediation strategies should be to develop certain kinds of innovative phytoreme-
diation, chemical and physical methods that have a potential to effectively reduce
both the amount and toxicity-level of the hazardous wastes. This broad area of
research requires applied field research, laboratory research and bench studies in
order to provide a scientific and technological base for management of hazardous
wastes in a cost-effective and efficient way, without producing secondary pollut-
ants. Heretofore, various types of soil remediation strategies have been suggested,
practiced, and tested to remove the soil pollutants. These include the use of metal-
guzzling plants, microbes, and animals. Phytoremediation is a green-phenomenon
which involves the use of plants and plant associated bacteria and fungi (endophytic
or rhizospheric), soil amendments (to improve soil physical properties, such as
structure, aeration, water permeability, retention, and drainage), and optimized
agronomic practices to remove soil pollutants. The use of plants as hyper-
accumulators of toxic metals is a simple, reliable, specific, affordable and eco-
friendly soil remediation approach (Macek et al. 2000; Rascio and Navari-Izzo
2011). Nearly, thirty-four plant families including five-hundred plant species that is
0.2% of the angiosperms worldwide have been identified as hyperaccumulators of
heavy metals (Zn, Cd, Pd, Cu, Ni and Co), metalloids (As) and non-metals (Se).
Most of the hyperccumulator-plants belong to the families asteraceae, brassicaceae,
buxaceae euphorbiaceae, flacourtiaceae, and rubiaceae (Reeves 2006). The different
criteria of the hyperaccumulator green-technology are (a) tolerance capacity, (b)
elimination efficiency based on plant biomass, (c) accumulation potential, (d) bio-
concentration factor (BCF) index and (e) translocation factor (TF) index (Verbruggen
et al. 2009). The use of natural and genetically engineered endophytic and rhizo-
spheric bacteria can synergize and accelerate the phytoremediation potential and
eliminate or beak-down soil toxicants (Glick 2003, 2010; Ma et al. 2015; Kong and
Glick 2017; Mesa et al. 2017). Although, hyperaccumulators of Ni, As, Mn, Se, Cd
and Zn have been described and experimentally proven but, the hyperaccumulation
of Cr, Ta, Cu, Co and Pb is still doubtful (van der Ent et al. 2013; Koubová et al.
2016). Various chemical, physical and thermal techniques have also been adopted to
enhance the rate of phytoremediation (Arlai et al. 2012; Veetil et al. 2014; Bashiri
et al. 2015; Xu et al. 2016). Chemical techniques that are deployed to extract the
pollutants from contaminated soils include chemical oxidation and solvent extrac-
tion (Ou-Yang et al. 2010). Chemical methods can lower the cost of remediation
process, as they can be operated on-site without any necessity to transport the soil
for off-site treatment. Furthermore, physical methods include soil vapour extraction
and soil washing (Fox 1996). Vapour extraction method leads to the vapourization
of the soil contaminants with the aid of pipes and wells in the soil. On the other
hand, thermal remediation techniques are currently being employed on highly con-
taminated soils (Falciglia and Vagliasindi 2015; Dupuis and Knoepfel 2015). This
method performs well where other methods may be costly to employ. Soils affected
by sewage wastes, industrial and mining effluents can be efficiently remediated with
the use of innovative biochar strategies (Anawar et al. 2015; Luo and Gu 2016).
Nanobioremediation is another emerging technology for remediation of pollutants
with the aid of biosynthetic nanoparticles. Because of unique chemical and physical
properties, nanoparticles have gained the attention of scientists. It is essential to
analyse and implement new remediation strategies which can upgrade the environ-
ment for sustainable livelihood. These strategies grouped under in-situ and ex-situ
remediation regimens need to be tailor-made for the environment under consider-
ation (Rani et al. 2013; Gomes et al. 2013; Cassidy et al. 2015; Kuppusamy et al.
2016). For this purpose, it is crucial to understand the efficiency and efficacy of
remediation technologies that are presently available (Eapen and D’Souza 2005;
Eapen et al. 2007; Stenuit et al. 2008; Kong and Glick 2017). It is crucial to analyse
the molecular mechanism and robustness of the degradation pathway of the toxic
compounds in plants or microbes before implementing it on a pilot scale. As men-
tioned before, some recent techniques like the use of biochar, biofilms and nanobio-
remediation have shown promise for the speedy biotransformation and sustainable
remediation of polluted soils and solid-wastes. The subsequent chapter focuses on
the available remediation techniques and their potential for soil remediation for the
period up to 2018 (Box 1.3) (Fig. 2.1).
1.5 Conclusions
Healthy land is an essential element in order to sustain human development and to

expand the livelihoods. But, with the boost of industrialization and urbanization, the
maintenance of soil quality and standard has become a global issue as the soil is a
vital medium of natural environment. Soil plays a major role as a medium for energy
and material transfer between atmosphere, hydrosphere and lithosphere. Soil
1.5 Conclusions 29
Box 1.3: “Swach Bharat Abhiyaan”

Swachh Bharat Abhiyan is also called as the ‘Clean India Mission’ or ‘Clean
India drive’ or ‘Swachh Bharat Campaign’. It is a national level campaign run
by the Indian Government to cover all the backward statutory towns to make
them clean. This campaign involves the construction of latrines, promoting
sanitation programs in the rural areas, cleaning streets, roads and changing the
infrastructure of the country to lead the country ahead. This campaign was
officially launched by the Honorable Prime Minister, Mr. Narendra Modi on
145th birth anniversary of the Late Mahatma Gandhi on second of October in
2014 at Rajghat, New Delhi, India.
It is a politics free campaign and inspired by the patriotism. It is launched
as a responsibility of each and every Indian citizen to make this country a
Swachh country. This campaign has initiated people globally towards the
cleanliness. Teachers and students of the school are joining this “Clean India
Campaign” very actively with great fervor and joy. Under this campaign,
another cleanliness initiative has been started by the UP CM, Yogi Adityanath
in March 2017. He has banned chewing paan, gutka and other tobacco prod-
ucts in the government offices all over UP.
As a part of the campaign, volunteers, known as Swachhagrahis, or
“ambassadors of cleanliness”, have promoted indoor plumbing and the
Community Approaches to Sanitation (CAS) at the village level. Other non-
governmental activities include national real-time monitoring and updates
from non-governmental organizations (NGOs) such as The Ugly Indian,
Waste Warriors and SWaCH Pune (Solid Waste Collection and Handling) that
are working towards the ideas of Swachh Bharat.
Water pollution is one of the biggest issues facing India right now. As may
be evident, untreated sewage is the biggest source of such form of pollution in
India. There are other sources of pollution such as runoff from the agricultural
sector as well as unregulated units that belong to the small-scale industry. The
situation is so serious that there is no water body in India that is not polluted
to some extent or the other. Rivers Yamuna, Ganga, Gomti, Ghaghara,
Chambal, Mahi, Vardha are among the most coliform polluted water bodies in
India. The coliform count must be below 104 MPN/100 ml and preferably
absent from water, to be considered safe for general human use, and for irriga-
tion where coliform may cause disease outbreak from contaminated-water
supplies to agriculture.
Industrial sector in India is witnessing rapid growth since the last decade
of twentieth century with reforms in economic laws and with establishment of
special economic zones (SEZ). Such rapid industrial growth has also increased
threat to the environment. In spite of great difficulty in its remediation in com-
parison with polluted air and water, soil pollution as a threat to human life is
by and large ignored at national level in India due to lack of comprehensive
(continued)
Box 1.3: (continued)

information on the subject. Though coordinated effort on assessment of soil
pollution is absent at national level, sporadic information has been generated
by several researchers on various aspects of pollution affecting the soil
quality.
Apart from Swach Bharat Abhiyaan, certain other rules/act are also in
action in India for the protection of environment resources. They include, the
manufacture, storage and import of hazardous chemicals rules 1989, biomedi-
cal waste (management and handling) rules 1998, the municipal solid waste
(management and handling) rules 2000, batteries (management and handling)
rules 2001, hazardous wastes (management, handling and transboundary
movement) rules, 2008 etc. These rules provide that the occupier shall be
responsible for environmentally sound and safe handling of any type of haz-
ardous substance generated during his/her establishment.
Societ y for environment and development has been carrying out ‘Mitti
Bachao Andolan’ (Save Soil Campaign) in five districts of Rajasthan (Kota,
Bundi, Jhalawar, Baran and Sawai Madhopur) India. Under this campaign,
farmers, students and school teachers are made aware about the soil and its
conservation and also equipped with portable soil testing kits during training
sessions to test the soil samples.
Soil Conservation Society of India (SCSI) was established in year 1952 at
Hazaribagh, Jharkhand, India on Dec 21, 1952. The headquarters of this soci-
ety is at New Delhi. This society was established with an objective to bring
awareness and knowledge about land resources of the country, also about the
conservation and care of soil and water. The objective of this society is sys-
tematic survey of relevant natural resources of the country i.e. climate, land,
water, plants, animals and human resources using the modern techniques of
remote sensing (computerized interpretation), storage and the retrieval of
information. It aims at the spread of knowledge regarding efficient soil and
water conservation and management practices including rainwater harvesting
and its use. The outreach (development projects) activities include soil and
water conservation practices for the effective control of soil erosion, sediment
discharge and floods, increasing the shelf-life of storage reservoirs for
irrigation.
The Indian Association of Soil and Water Conservationists (IASWC) was
established in 1972 at Dehradun, India as a non-profit body to encourage land
use for conservation of soil and water resources to sustain mankind. It also
promotes and disseminates knowledge, practical skills and research in the sci-
ence and technology, for soil and water conservation.
References 31
pollution is caused by natural sources such as volcanic eruptions, earthquakes etc.

and anthropogenic activities such as industrialization, urbanization, agricultural
practices, nuclear technology, mining and smelting industries etc. Human activities
disturb the balance between different ecosystems and shrink the agricultural land
which affects the food security.
The two most important categories of soil pollutants are heavy metals (Hg, Zn,
Pb, Cd, As, Cu) and organic compounds (PAHs, PCBs etc.). Different types of
industrial wastes, manures, fuel combustions are the main sources of these pollut-
ants. Presences of these contaminants above the threshold level cause toxicity to all
living beings. Most of these contaminants have no biological role but have gained
the attention of scientists due to their toxicity, persistence, abundance and accumu-
lation in the environment. It has been reported that the area containing polluted soils
has reached up to 2250 million hectares and is further expanding (Yao et al. 2012)
This alarming rise in the soil pollution has forced the scientists to develop vari-
ous strategies for its remediation and reclamation. However, the present environ-
mental policies lack clear commands and hence do not address the global concern
of soil pollution properly. Besides that, practical implementation of these policies is
not beneficial due to non-operational soil quality standards, uncertain soil remedia-
tion methods, limited funding, poor supervision and technical capabilities of reme-
diation industries. In addition, environmental policies regarding the management of
soil standards should focus on the implementation of appropriate and integrated soil
remediation strategies in order to improve the soil quality.
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Chapter 2
Ex situ Soil Remediation Strategies
Abstract Ex situ techniques involves the treatment of contaminated soil, away

from the polluted site. Ex situ bioremediation can be operated in two ways which
include solid phase bioremediation and slurry phase bioremediation. Besides its
high cost, these techniques are highly efficient, easy to control, faster and have great
potential to treat large number of contaminants from the soil. Ex situ techniques
involve land farming, biopile, windrow, soil washing, composting, bioreactor, ion
exchange, adsorption/absorption, pyrolysis and ultrasound technology. These tech-
niques can be successfully applied for the treatment of fuel hydrocarbons, haloge-
nated and non-halogenated organic compounds as well as for various pesticides.
Keywords Ex situ · Composting · Bioreactor · Land farming · Pyrolysis
2.1 Introduction
These techniques involve the treatment of polluted soils away from the polluted
area. The success of ex situ bioremediation techniques depends on the (a) cost
incurred, (b) depth of the pollutant, (c) type of the pollutant, (d) geographical loca-
tion of the pollutant, (e) degree of pollution and (f) the choice of ex situ technique
used (Philp and Atlas 2005). The ex situ strategies can be grouped under solid-phase
bioremediation (land farming, soil biopile, windrow, windrow-composting) and
slurry-phase bioremediation (bioreactors). Figure 2.1 provides a general summary
of the ex situ and in situ remediation techniques. Although, solid-phase bioremedia-
tion is easy to maintain, but it requires much space and the process of decontamina-
tion is less time-effective than slurry-phase bioremediation. The ex situ remediation
treatments are faster, manageable and can successfully remediate several inorganic
and organic pollutants (Prokop et al. 2000). Following are the types of ex situ
strategies:

40 2 Ex situ Soil Remediation Strategies
Fig. 2.1 Ex situ and in situ remediation strategies for treatment of polluted soil
2.2 Land Farming
It is one of the simplest bioremediation techniques as it requires less capital and

expertise. It may be practiced both in situ and ex situ depending upon the depth of
the polluted zone. If the excavated polluted soils are treated on-site then the treat-
ment is said to be in situ and vice-versa. In other words, it can be said that when a
pollutant lies less than 1 m below the soil surface, bioremediation may not require
excavation. On the contrary, if the pollutant lies more than 1.7 m below the surface,
then it is brought to the surface for effective soil remediation mediated by autoch-
thonous microorganisms (Nikolopoulou et al. 2013; Volpe et al. 2012; Silva-Castro
et al. 2015). In land farming optimal aeration, nutrients and irrigation enhance the
activity of microorganisms which further enhances the process of bioremediation. It
is usually used for the remediation of PAHs (Silva-Castro et al. 2012; Cerqueira
et al. 2014). Being a simple technique, it can be implemented to remediate large
amount of contaminated soils with less energy input (Maila and Colete 2004).
However, this treatment can be practiced in any location and climate, adhering to
the governmental policies (Besaltatpour et al. 2011). Although, land farming is a
feasible technique but, it is slow and not much effective in comparison to the other
ex situ bioremediation treatments.
Drawbacks of this technique include the requirement of a large processing area,
unavoidable environmental conditions hampering microbial activity, cost of excava-
tion, and inefficiency in the removal of inorganic pollutants as well as toxic volatiles
(Maila and Colete 2004).
2.3 Biopile
As the name indicates, this method involves piling or stacking of contaminated soils
to facilitate microbial activity through cost-effective and controlled nutrient amend-
ment, aeration and irrigation practices (Fig. 2.2) (Whelan et al. 2015). Thus, the
2.4 Windrow 41
Fig. 2.2 Biopile set up for treatment of polluted soil
biopile technique can prevent volatilization of contaminants with low molecular

weight and finds application in remediation of polluted soils of very cold regions
(Dias et al. 2015; Whelan et al. 2015). The advantage of a biopile setup is that a
large amount of contaminated soil can be treated in a less area. The pretreatment
(sieving, aeration) of contaminated soils enhances the process of biopile-mediated
remediation (Delille et al. 2008). Other amendments such as straw, saw-dust, wood-
bark and wood-chips do enhance the process of biopile set-up (Rodrıguez-Rodrıguez
et al. 2010). However, the running of the biopile set-up and its maintenance add to
its cost. Moreover, the heated air that is generated tends to dry the soil which further
inhibits the microbial activity and promotes volatilization of contaminants rather
than biodegradation (Sanscartier et al. 2009).
2.4 Windrow
Windrow is another ex situ soil remediation technique which is a modification of

biopile treatment. Windrow treatment involves frequent whirling of piled polluted
soils to enhance the bioremediation process (Fig. 2.3). This periodic turning of pol-
luted soils along with irrigation enhances aeration, microbe-mediated degradation,
uniform distribution of pollutants and thus, accomplishes bioremediation through
assimilation, biotransformation followed by mineralization (Barr 2002). In com-
parison to biopile technique windrow ensures higher hydrocarbon removal, the rate
of which depends on the soil type (Coulon et al. 2010). However, this technique may
not be successful if the polluted soil contains toxic volatile compounds or releases
methane (greenhouse gas) during the process (Hobson et al. 2005).
Fig. 2.3 Windrow set up

for treatment of polluted
soil
2.5 Composting
Composting is a technique which is used to degrade and convert the organic pollut-
ants and has been practiced for remediation of organic toxicants, for the last
3–5 years (Lukić et al. 2017) (Fig. 2.4). The investigated pollutants include mono-
aromatics (toluene and benzene), pesticides [2,4-dichlorophenoxyacetic acid (2,4-
D) and diazinon], petroleum hydrocarbons, explosives [2,4,6-trinitrotoluene
(TNT)], polycyclic aromatic hydrocarbons (PAHs) [phenanthrene, anthracene,
benzanthracene, benzopyrene] and chlorophenols [pentachlorophenol (PCP)].
Bastida et al. (2016), considered in a model experiment the effects of petroleum
contamination and bioremediation potential of manure, as an enhancer of microbial
mobility in semiarid soils. The addition of compost in polluted soil enhanced the
removal of PAHs and alkanes (88%) after 88 days, in a goldmine site, in Iperindo,
Osun State and three industrial locations of Ogun State (Taiwo et al. 2016). A higher
degradation percentage of total hydrocarbons (~82%), polycyclic aromatic hydro-
carbons (~93%), and n-alkanes (~ 96%) was observed on treating with 75% of
organic wastes (Paladino et al. 2016). Compost technology, in association with bio-
stimulation and bioaugmentation has been successfully used for the treatment of
polluted sludges and soils. High microbial concentration, lower hydrocarbons and
salt concentration, proper nutrients and water supply can successfully enhance the
biological activity, and ultimately enhance the biodegradation rate (~75%) of
organic wastes. This technique has also been used in combination with land farming
or windrows which are more promising for efficient and economical removal of
contaminants from soil (Lukić et al. 2017).
2.6 Soil Washing 43
Fig. 2.4 Soil composting for treatment of polluted soil
2.6 Soil Washing
Soil washing is a simple technique to remove the contaminants that adhere to the
surface of the soil particles, in an aqueous-based system (Fig. 2.5). They are then
extracted from polluted soils by dissolving or suspending the polluted soil in wash-
ing solution. These contaminants (organic and inorganic) then bind to silt, clay and
organic soil particles, chemically or physically. Most of the clay and silt adhere to
the larger particles like gravel and sand. The washing treatments break adhesive
bonds between larger and smaller particles and thus separate them. Thus, concen-
trating the contaminants into small volume on the basis of gravity separation, attri-
tion scrubbing and particle size separation makes their disposal more feasible.
Soil washing has been successfully used for remediation of soils that were con-
taminated with heavy metals, selected VOCs, SVOCs, fuels, explosives, and pesti-
cides (Mann 1999). Compared to US the technique of soil washing has been
deployed extensively in Europe. Although, soil washing is a cost-effective versatile
and well-established technique but, performs well on coarse-particle and sandy
soils having little or no organic matter. Moreover, the complex mixtures of pollut-
ants may require several washings.
Different types of additives like surfactants, acids and chelating agents are mixed
with the soil so as to solubilize the contaminants present in it. Most commonly,
those surfactants are added which have both hydrophilic and hydrophobic proper-
ties. This amphoteric nature of surfactants enhances the solubility of hydrophobic
and hydrophilic organic contaminants (Mao et al. 2015). Different categories of
surfactants such as anionic, cationic, non-ionic and zwitter-ionic have been success-
fully used for remediation of polluted soils (Fountain et al. 1996). Table 2.1 shows
various ionic and non-ionic surfactants used for the remediation of soil contami-
nated with organic and inorganic pollutants. The addition of surfactants leads to
various types of interactions: hydrophilic groups enters into the aqueous phase
while the lipophilic groups combine with hydrophobic pollutant molecules present
Fig. 2.5 Soil washing set up for treatment of polluted soils
in the soil. These dissolved pollutants have better mobility and can be removed or
extracted from the soil (Kim et al. 2011).
Heavy metal particles can be successfully removed from the soil via complex
formation (Ochoa-Loza et al. 2001) with surfactant molecules and via ionic
exchange (Swarnkar et al. 2012). In a study, nonionic surfactant (Ammonyx KP),
cationic surfactant (DPC) and ionic biosurfactant (JBR-425) were analysed for their
metal removal efficiency. It was observed that among them JBR-425 was effective
for the removal of 56% Cu, 39% Zn, 68% Pb and 43% Cd from contaminated soil
(Slizovskiy et al. 2011). Various radionuclides such as Cs137 (caesium), Sr90
(strontium) and U238 (uranium) can also be removed from the contaminated soil
via complex formation, dissolution and ion exchange methods (Gadelle et al. 2001;
Willms et al. 2004). Hexadecyl trimethyl ammonium, a cationic surfactant has been
successfully used for the treatment of Cs+contaminated soils (Evans 2003).
Biosurfactants are the important surface active biological compounds which are
produced in living organisms (Paria 2008). Biosurfactants can be ionic or anionic in
nature and the most commonly used biosurfactants are lipo-peptide (polymyxin,
surfactin), humic substances and glycolipid (fructose lipids, rhamnolipids, sophoro-
lipids) (Salati et al. 2011). The hydrophobic parts of biosurfactants are made from
aliphatic hydrocarbons while the hydrophilic parts are made from phosphates,
cyclic peptides, polysaccharides, carbohydrates and amino acids (Mulligan 2005;
Rahman and Gakpe 2008; Pacwa-Plociniczak et al. 2011).
Moreover there are various factors which should be taken into consideration for
the successful implementation of surfactant enhanced remediation such as solubil-
Table 2.1 Application of various ionic and non-ionic surfactants for the remediation of soil contaminated with organic and inorganic pollutants
Soil source/
Surfactant Contaminated sites Major contaminants Results References
Ionic
Dowfax, sodium dihexyl Alameda Point Naval DNAPL, especially TCA and TCE 95% DNAPL (dense non-aqueous Strbak (2000)
2.6 Soil Washing
Sulfosuccinate, NaCl and CaCl2 Air Station Site, phase liquid) removal and 93%
Alameda, CA surfactant recovery
4 wt% Isalchem 123 (PO)7.7 sodium Millican Field, Pearl Petroleum, LNAPLs 87.5% of the LNAPL (light non-
ether sulfate with 8% SBA Harbor, Hawaii aqueous phase liquid) in soil was
(Secondary butyl alcohol) cosolvent recovered
AOT/Calfax 16 L-35 Underground storage Diesel fuel, and gasoline fuel 75–99% benzene reduction, 65–99% Shiau et al.
tank site in Oklahoma NAPL TPH (Total petroleum hydrocarbon) (2003)
reduction
Mixture of Alfoterra 123–4-PO Chevron Cincinnati BTEX (benzene, toluene, LNAPL reduced from 8% to less than USEPA
sulfate, 8% 2-butanol, Emcol-CC-9 Facility in Hooven ethylbenzene and xylene), LNAPLs 1% residual saturation (2005)
and calcium chloride
50 ml HTAB (Hexadecyl trimethyl Nankai University, Aldicarb (carbamate pesticide) 56% desorption ratio of aldicarb Xu et al.
ammonium bromide) Tianjin, China (2006)
1.5% (w/w) CTAB (Cetyl Clay soil from Benzene series, naphthalene and Organic pollutants were removed by Ranjan et al.
trimethylammonium bromide) Meachitoba Province, phenanthrene 98.9% (2006)
Canada
10−2 M SDS (Sodium dodecyl Agricultural soil from Cd(II) After 18 days 94% removal efficiency Giannis et al.
sulfate) Crete island, Greece of Cd (2007)
10 wt% NaCl +4 wt% SDS Organic contaminated 1,2,4-trichlorobenzene (TCB) 97% removal efficiency for TCB Lee et al.
soil in Pyeongtaek, (trichlorobenzene) (2008)
Korea
(continued)
45
46
Soil source/
8 mM SDS Fuel oil contaminated Diesel 97% removal efficiency for diesel Khalladia
soil near Algiers, et al. (2009)
Algeria
Spolapon AOS 146 solution An incinerator plant in PCBs 56% efficacy for PCBs Svab et al.
Czech Republic (polychlorinated biphenyls) (2009)
decontamination
20 ml 0.5% Texapon-40 Heavy metals Heavy metals like Cd, Zn, Cu, Ni Cd, Ni and Zn were removed by Torres et al.
contaminated soil from 83.2%, 82.8% and 86.6% respectively 2012)
a metallurgy plant,
Mexico
Non-ionic
0.5% Canarcel Tween 80 An out-of-service oil Polluted by gasoline and diesel, Average TPH removal was 87.1% Iturbe et al.
distribution and storage like MTBE (Methyl tert-butyl (2004)
station, Mexico ether) benzene, toluene, BTEX
Tergitol NP-10 Heverlee bos’ after Diesel Diesel oil was removed by 50% Vreysen and
2
removal of the surface Maes (2005)

litter, Belgium
2% sorbitan monooleate Soil from a pilot-scale Diesel, coal oil and lubricating oil 88% of total petroleum hydrocarbon in Lee et al.
site in Pusan, Korea soil was removed (2005)
6% Tween 80 Bachman Road site in Tetrachloroethylene Washing out 19 L tetrachloroethylene Ramsburg
Oscoda, northeast et al. 2005
Michigan, US
25 ml 0.05 M Brij-35 Soil prepared in the Hydrophobic aromatic compounds 43%–69% removal efficiency Fabbri et al.
laboratory (2008)
Ex situ Soil Remediation Strategies
Soil source/
A combination of 0.213 M MGDA An old wood PAH (Polycyclic aromatic Concentrations of As and PAH were Elgh-Dalgren
(methylglycinediacetic acid) preservation site in hydrocarbons) and As reduced from 105 ± 4 and et al. (2009)
Elnaryd, south east 27 ± 0.7 mg/kg to 25 and 10 mg/kg
Sweden after washing, respectively
Tween 80 Loamy sand in Madrid, P-cresol >70% extraction efficiency Rosas et al.
Spain (2011)
2.6 Soil Washing
20 ml 0.5% Tween 80 Heavy metals Cd, Zn, Cu, Ni, Pb and other heavy Removal efficiency was 85.9% Cd, Torres et al.
contaminated soil from metals 85.4% Zn and 81.5% Cu (2012)
a metallurgy plant,
Mexico
0.15% (v/v) Empilan KR6 and Crude oil contaminated Petroleum hydrocarbon TPH removal increased from 15% to Couto et al.
1.6 mg/kg bioaugmentation soil in refinery 28% (2012)
commercial product MicroSolv-400
47
ity of surfactants towards targeted pollutants, adsorption of surfactants in soil and

biodegradability/toxicity of the surfactants (Mao et al. 2015). Biological and physi-
cochemical characteristics such as hydraulic conductivity (Renshaw et al. 1997) of
soil may get altered due various types of surface reactions between surfactant mol-
ecules and soil particles (Jia et al. 2005). Toxic or non-biodegradable surfactants
pose hazardous effects on soil ecosystem. Certain anionic surfactants have the abil-
ity to bind with biological molecules such as DNA, enzymes, peptides etc. and can
change the biological functioning of organisms (Cserhati et al. 2002). While, the
cationic and nonionic surfactants can affect the bacterial cell membrane (Evankovic
and Hrenović 2010) and exhibit anti-microbial activity (Santonicola et al. 2008).
2.7 Bioreactor
Bioreactors have now become one of the best ex situ strategies for treatment of pol-
luted soils containing recalcitrant pollutants, under controlled/automated environ-
mental conditions (Robles-González et al. 2008; Das and Chandran 2011). They are
often used to treat the polluted soils in slurry or solid phases. Use of bioreactor for
soil remediation has various advantages over other ex situ bioremediation treat-
ments. Total control over the bioreactor parameters such as substrate and inoculum
concentrations, pH, temperature, agitation and aeration rates are the major benefits
of bioreactor-based bioremediation (Uddin et al. 2017). Bioreactors have been
reported to successfully remediate the soils contaminated with volatile organic
compounds (VOCs) such as toluene, benzene, xylenes and ethylbenzene. They are
often used to determine the actual potential and feasibility of biological techniques
in the remediation of soil and water (Ioannou-Ttofa et al. 2017). In slurry-based
system the reduction rate of pollutant depends mostly on the degradation rate of
microbes accessible in the system and the outcome represents the remediation
potential of microbes (Robles-González et al. 2008). Su et al. (2012), evaluated the
bioremediation of river sediments polluted with 9% phenanthrene. Under optimal
conditions, 22–63% phenanthrene was removed within 30 days of treatment with
dehydrogenase and polyphenol oxidase enzymes. Wick et al. (2011), reported the
remediation potential of pristine sediment contaminated with anthracene, fluoran-
thene, phenanthrene, and pyrene. Rahman et al. (2017), characterized the microbial
population in the thiocyanate degrading bioreactor that includes solid particulate
tailings of mine water. Many organisms have containing genes for denitrification
and sulfur oxidation have been isolated, but only Thiobacillus sp. has thiocyanate
degradation genes. Nguyen et al. (2017), used biofilm reactor systems to evaluate
the Pb removal capacity of an indigenous association of five sulfate-reducing bacte-
ria (SRB): Desulfomicrobium salsugmis, D. autotrophicum, D. escambiense,
Desulfovibrio carbinolicus and D. vulgaris. It has been reported that under con-
trolled bioreactor conditions, microbial consortium effectively degraded the marine
fuel (MF-380) and the effect of variables such as pH, temperature and C: N ratio
was also analyzed (de Almeida et al. 2017). Bioreactors hold a promising potential
2.9 Ion Exchange 49
to degrade royal demolition explosive (RDX), trinitrotoluene (TNT), chlorobenzene

and dichloro-benzene and other organic contaminants of industrial areas including
leaking storage tanks. Genetically modified microorganisms can be deployed for
enhancing the process of bioreactor-based bioremediation (bioaugmentation) and
before transporting the treated soil to the field the organisms can be destroyed, so as
to prevent horizontal gene transfer of the foreign gene. However, there are certain
constraints which hinder the performance of bioreactors which include high con-
centration of toxic pollutants, low temperature, some problematic microbes and dis-
posal of discharged effluents.
2.8 Absorption/Adsorption
Different types of absorbents and adsorbents have been frequently used for reme-
diation of contaminated soils. Rate of adsorption or absorption of contaminants can
be enhanced by using certain surfactants (inorganic). These surfactants start such
kind of reactions (surface tension diminishment, solvency upgrade, micellar solubi-
lization, wettability and frothing limit) which assist the extraction of pollutants
(Mulligan et al. 2001). Combined use of surfactants (ionic, cationic and non-ionic)
and other complexing agents (diphenylthiocarbazone and EDTA) have been tested
for removal of Pb, Cd, Zn from soils polluted with heavy metals (Doong et al. 1998).
Zhang et al. 2007, effectively expelled Pb from polluted soil by washing it with
EDTA after SDS treatment. Commercially activated carbon has also been widely
used for adsorption of Sb and Ar from contaminated soils (Rajaković 1992; Lorenzen
et al. 1995; Navarro and Alguacil 2002).
2.9 Ion Exchange
Ion-exchange is a technique which involves exchange of anions and cations between

the media and contaminants. The materials which are generally used for ion-
exchange are resins (natural polymers) with functional groups attached over it
(FRTR 2012). When liquids are passed over these resins, cations and anions of
resins and pollutants are exchanged with each other. This method can successfully
extract the nitrates, silicates, dissolved metals, ammonia and radionuclides from
aqueous medium (Vilensky et al. 2002; Rengaraj et al. 2003). Ion exchange resins
are environmentally compatible and highly reliable as they are insoluble and can be
used for long time in majority of the cases. This technique has been used from late
1990s in the industries and is now available commercially (Harmer and Sun 2001;
Mergen et al. 2008). The potential of resin membrane increased with electrokinetic
soil was studied for metal removal and it has been observed that ~96% of Pb and Cd
were removed by this method, within 4 days (Kim et al. 2005). A study was con-
ducted at Thala valley tip, Antarctica and it was observed that the resins chelating
with iminodiacetic acid can help to reduce about 92–100% of heavy metals within
a year (Woodberry et al. 2007). The limitation of this technique is that oil, grease,
and other organic and inorganic compounds which are present in the treated media
can block or damage the resin (Beril Gonder et al. 2006). Waste water, which is
produced as an outcome of resin treatment requires additional disposal and treat-
ment which adds to the cost of this method (Pintar et al. 2001). There are some other
concerns also with respect to noise generation, treatment size, odor and the disposal
of used resins (Srinivasan and Sorial 2009). Most of the resin beds can be reused
while some resins can be used only once (Alexandratos 2008). Although, ion
exchange system is efficient but is not free from complications.
2.10 Pyrolysis
Pyrolysis is a promising treatment for soil remediation in which the chemical

decomposition of toxic compounds occurs by the use of high temperature (>430 °C),
high pressure and anoxic conditions (Venderbosch and Prins 2010) (Fig. 2.6). This
treatment transforms the pollutants or toxic materials into organic liquids, water,
char and fuel gas (Rofiqul Islam et al. 2008). Organic and inorganic contaminants
for e.g. PCBs, SVOCs, PAHs, paint wastes, cyanides, refinery wastes, pesticides,
wastes from processing of synthetic rubber, dioxins, mercury, soils contaminated
creosote and hydrocarbons, wastes from coal-tar and wood-treatment and radioac-
tive wastes are most commonly remediated by pyrolysis treatment (Mohan et al.
2006; Arvanitoyannis et al. 2007). The equipments used in pyrolysis consist of rota-
tory kilns, fluidized-bed furnaces and molten salt destructions. This is a reliable
technology which ensures a high flexibility in its design and operation, high
Fig. 2.6 Schematic representation of Pyrolysis process

2.11 Ultrasonic Technology 51
pollutant-cleaning efficiency, no CO2 production, effective reduction in weight and

volume of the toxic materials, low operational and maintenance cost, stable opera-
tion for organic and inorganic wastes, energy production opportunities and efficient
material recovery and feasibility of implementation in both small and large-scale
(Inguanzo et al. 2002; US EPA 2012). However, this technique is not free from limi-
tations. The increased soil moisture content adds to the cost of treatment and certain
gases such as CO, H2, CH4 that are released during the course of operation are also
a concern. Moreover, the major limitation of this technology is its feasibility in
treating organic pollutants from oily sludges and soils compared to inorganics.
A report on Cr(VI) remediation from its ore processing residue highlighted
99.1% success (Zhang et al. 2009). In a similar report, Thuan and Chang (2012)
could remove 90% pentachlorophenol (PCP) within 40 min (pyrolysis temperature
350 °C). Debela et al. (2012), analysed that mixing of heavy metal (Cd, Zn and Pb)
contaminated soil with 5–10% of sawdust followed by pyrolysis at 400 °C can effi-
ciently retard the concentrations of heavy metals. Interestingly, pyrolysis is an
excellent technique for the disposal of phytoremediation crops. In a report by Stals
et al. (2010), the Pb and Cd phytoaccumulator plants were subjected to this eco-the
friendly technique which economically yielded oil (as source of fuel) and char (as
soil amendment).
2.11 Ultrasonic Technology
This technology involved the use of high temperature up to 4726 °C, high frequency
up to 18 kHz and high localized pressures up to 50 MPa which degrades the con-
taminants because of chemical reactions (Adewuyi 2001). This technology is more
valuable as it requires less area, less energy requirements, less installation and
maintenance cost (Thangavadivel 2010). This treatment has been successfully used
for remediation of soil polluted with petroleum hydrocarbons, pesticides, chlori-
nated solvents and heavy metals. It has also been used at a large scale for industrial
uses such as audible frequency sonication of 100 Hz was used in Canada and low
frequency ultrasound were used in Australia for treating the polluted soil (Mason
et al. 2004). It has been reported that ultrasonic technology was successfully used
for the desorbing or leaching the contaminants from soils and sediments (Kazi et al.
2006; Abramov et al. 2013). In a report by Mason et al. (2004), it has been found
that low frequency ultrasound efficiently increased the rate of soil washing treat-
ment. Ultrasound technique and biomass obtained from transgenic algal species
Chlamydomonas reinhardtii has been successfully used in combination for efficient
removal of heavy metal (Hg) from polluted sediments (He et al. 2011). In a study by
Thangavadivel 2010, contaminated slurries treated with anionic detergent sodium
dodecyl sulphate i.e. SDS, supplied with low frequency and high power ultrasound
energy were used to enhance the extraction of DDT from polluted soil. Combined
action of ultrasound energy and Fenton oxidation reactions were tested for treating
the organic pollutants present in contaminated soil. It has been found that this
combination effectively removed 87% and 92% of xylene and toluene respectively
from the polluted soil, within 2 days of treatment (Flores et al. 2007; Huguenot et al.
2015). It has also been reported by Pham et al. 2009, that eletrokinetics remediation
treatment significantly enhanced the removal rate (for phenanthrene and hexachlo-
robenzene) up to ten times when used in combination with ultrasound energy.
There are also some limitations of this treatment such as, during the ultrasound
desorption treatment, repeated power cuts due to the increased frequency hinder the
rate of remediation. Finer particles take longer time for removal as compare to large
particles because more sonication time is required for small particles. No doubt,
ultrasound desorption is a promising and reliable treatment and an alternative to
solvent flushing or other conventional method but further elaboration is required for
its large-scale efficiency.
2.12 Conclusions
Major advantage of ex situ treatment over in situ treatment is that it requires less
time period to operate and the byproducts are not released in the environment until
they are converted to non-harmful products. These strategies can successfully be
applied for the treatment of polluted soils, groundwater and sludges polluted with
pesticides, petroleum hydrocarbons, and other organic and inorganic pollutants.
However, the main limitations of these treatments are the removal and transporta-
tion of contaminated soil to the treatment area which eventually leads to its high
cost. Moreover, the emission of substances during the remediation treatment may
also affect the surrounding environment. The above limiting factors should be taken
into consideration to improve the large-scale application of the aforementioned
techniques. The ex situ treatments can also be used in an integrated manner and with
other in situ and bioremediation strategies which will surely enhance the rate of
bioremediation when deployed on a pilot scale.
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Chapter 3
In Situ Soil Remediation Strategies
Abstract Physical remediation of polluted soil is one of the in situ remediation

strategies, generally involves disintegration of the contaminants by methods such as
evaporation, heating or washing. These methods are based on the principles used for
the extraction of desired metal from their respective mineral ores. Suitability and
efficacy of physical separation technique depends upon type of the soil, shape, dis-
tribution and size of the contaminant, amount of humus, moisture and clay in the
soil. These techniques can be used for the extraction/removal of both inorganic and
organic contaminants from polluted soils and involves low operational cost. The
selection of the most suitable physical treatment largely depends on the type of
contaminated soil and type of the contaminant (organic or inorganic). Most com-
monly these techniques are applicable to the soils of industrialized urban areas
(contaminated through anthropogenic activities). The present chapter discusses
various physical methods of soil remediation including physical separation, soil
flushing, volatilization, froth flotation and thermal heating.
Keywords Physical separation · Soil flushing · Thermal remediation · Froth

flotation
3.1 Introduction
As the name indicates, these techniques involve the remediation of soil contami-
nated with inorganic and organic pollutants, at the site of pollution. The in situ
techniques are most commonly used to treat chlorinated solvents, heavy metals,
hydrocarbons and dyes deploying (single or in combination) microbes, plants, phys-
ical and chemical methods (Folch et al. 2013; Kim et al. 2014; Frascari et al. 2015;
Roy et al. 2015). The status of electron acceptor, nutrient availability, moisture con-
tent, temperature, pH and adequate soil porosity are the most important require-
ments that are crucial for in situ bioremediation to be successfully accomplished
(Philp and Atlas 2005). This chapter provides a summary on different in situ

60 3 In Situ Soil Remediation Strategies
Fig. 3.1 Comparison of physical, biological and chemical methods of soil remediation
strategies for remediation of polluted soils. These strategies can be categorized into
five major divisions based upon the type of the treatment:
• Biological methods
• Physical methods
• Chemical methods
• Biochar/biofilms
• Nanobioremediation
Biological methods include the use of bacteria, algae, fungi, plants and animals
so as to degrade or immobilize the contaminants in the soil. Physical methods con-
sist of physical separation, soil flushing, volatilization, froth flotation and thermal
treatments. While chemical methods comprises of leaching, fixation and electro
kinetic remediation (Fig. 3.1). Carbon sequestration, nutrient exchange, water hold-
ing, absorption/adsorption and oxidation/reduction are summarized under biochar/
biofilm. Nanobioremediation is also an emerging technology for soil remediation.
Each in situ strategy has its own advantages and limitations which depend upon the
soil condition, characteristics and pollutant content.
3.2 Physical Methods of Soil Remediation
Physical remediation techniques involve both, soil-replacement and thermal desorp-

tion methods (Su 2014; von Lindern et al. 2016). Soil-replacement is further of
three types i.e. soil spading, soil replacement and soil importing. Soil spading
includes digging of contaminated soil and spreading the contaminants into deep
3.2 Physical Methods of Soil Remediation 61
sites; soil replacement is the removal of polluted soil while, soil importing is the
addition of unpolluted soil into an area of polluted soil to dilute the concentration of
contaminants. The soil replacement methods can efficiently isolate the contami-
nated soil and decrease the effect of pollutants on the environment. Apart from this,
the technique is costly and is applicable only for small areas which are severely
polluted (Ahmad 2016).
The thermal desorption method is based on the pollutants’s volatility and deals
with heating the polluted soil with microwave, steam and infrared radiations to vol-
atilize the contaminants (volatile heavy metals like Hg and As). These volatile metal
contaminants are then accumulated using the vacuum negative pressure or carrier
gas (Li et al. 2010). Traditionally, thermal desorption technique can be categorized
into low temperature (90–320 °C) and high temperature desorption (320–560 °C).
Besides that, there are some limitations in this technique which include long desorp-
tion time and the use of expensive devices (Aresta et al. 2008).
The efficiency of physical methods depends upon various soil properties such as
size, shape and dispersal of particles, clay, moisture and humus content, hydropho-
bic and magnetic properties of the soil particles. Besides that, physical treatments
are not applicable in following conditions:
• When the soil-clay or slit content is more than 30–50%.
• When metal contaminants are tightly bound to soil particles.
• When there is high humic content.
• When there is much similarity between the properties of soil particles and metal
bearing particles.
Physical remediation involves the following methods:
3.2.1 Physical Separation
Physical separation is generally considered as particle separation process and is

mostly applicable when heavy metal contaminants are of a particular size. It involves
the techniques which are generally used in mineral processing and mining indus-
tries to extract the desired metals from mineral ores (Dermont et al. 2008). In case
of soil remediation, these technologies have been successfully used to separate
heavy metals from contaminated soils (Williford and Bricka 2000; Mercier 2000).
There are two types of physical separation techniques: (a) magnetic separation –
The soil particles have some magnetic properties which can vary from ferromag-
netic (largely positive), paramagnetic and organometallics (intermediate) to organic
(negative) (Rikers et al. 1998). Magnetic field of low intensity can attract the ferro-
magnetic substances, while a high intensity magnetic field can attract the paramag-
netic substances (Mercier et al. 2001). Low intensity magnetic fields have been
successfully used to regain the spent amunition debris at military sites. High inten-
sity magnetic field can be applied for the elimination of heavy metals such as Zn,
Cu, Cr, Ni, Pb from polluted soils (Rikers et al. 1998). However, in case of
heterogeneous soil matrix, the magnetic separation is not feasable for removal of
heavy metals (Mercier 2000); (b) electrostatic separation – It is rarely used for soil
remediation. Lead-based paint chip separation and recovery technology is one such
example of electrostatic separation being practiced at full scale, in the Pittsburgh
Mineral and Environmental Technology Inc. (Gilbert and Weyand 1990). This tech-
nique efficiently removed 70–80% PAHs, 36–40% Pb, 61–65% Cu and 27–33% Zn
from contaminated sandy beach sediments in Canada (Veetil et al. 2014). However,
further research is required for its full-scale application.
3.2.2 Soil Flushing
Soil flushing involves injection or infiltration of a mixture of water, acidic or basic

solutions, chelating agent, reducing agents, and surfactants into the contaminated
area, followed by mobilization of contaminants by emulsion formation after a
chemical reaction with the flushing solution. Soil flushing setup involves injection
well to inject the solution and extraction well to extract the solution containing the
contaminants. These contaminants are then further treated to transform the contami-
nants into less toxic form (Fig. 3.2). The success of soil flushing technique relies on
the (a) nature of contaminants being processed, (b) soil characteristics (homoge-
neous and non-homogeneous), (c) ability to inject the solution, (d) control of the
Fig. 3.2 Soil flushing method for removal of soil contaminants

injection flow and (e) the recovery of flushing solution. Water is preferred as a flush-
ing liquid to extract the hydrophilic constituents from polluted soils. Acidic solu-
tions aid in recovery of metals or basic organic materials while, basic solutions can
be used for metals such as zinc, tin, lead and some phenols. For the recovery of
some heavy metals, chelating (EDTA), complexing (Cl−, CN−) and reducing (H2,
CO, Fe) agents are also used. Surfactants aid in the recovery and removal of hydro-
phobic organics while cosolvents are effective in removal of some organics (USEPA
1991).
Surfactants (cationic, anionic, non-ionic) are most commonly used in soil flush-
ing for removal of fuels and chlorinated solvents. Among the aforementioned cate-
gories anionic or non-ionic surfactants are preferred as they are generally less toxic
and their negative or neutral charge reduces the chances of their sorbtion to nega-
tively charged clays. These surfactants can be mixed with a cosolvent (e.g. isopro-
panol) which increase the solubility of surfactant in solution and thus provide
surfactant/contaminant solution an acceptable viscosity. Later, the cosolvent isopro-
panol is recovered during the recovery process owing to its toxicity and persistence.
Therefore, ethanol has now been substituted for isopropanol (NAVFAC 2002). A
homogeneous soil system greatly reduces the remediation time. There are certain
factors which limit the applicability and efficiency of the flushing method. The
adhesion of certain surfactants molecules reduces the soil porosity, while the hetero-
geneous soils with low permeability makes the remediation process cumbersome.
Furthermore, some soil factors like (a) high cation exchange capacity, (b) high buff-
ering capacity, (c) high organic content, and (d) pH adversely affects and increases
the remediation time which further adds to the cost of treatment.
3.2.3 Volatilization
Semi-volatile and non-volatile organic contaminants present in soil can be removed

by heating the soil at a temperature required to vaporize the pollutants. Vapourization
of contaminants can be achieved by steam, aeration, vacuum, and heating the soil to
several hundred-degree centigrades. Among these, aeration of contaminated soil is
a cost-effective, simple and convenient process. Volatilization of pollutants can be
accelerated by heating the contaminated soil in a container. Steam-induced volatil-
ization involves the heating of polluted soil and condensation of steam below
100 °C. The wet solids from the degrading operations are slurried with water and
then steam is passed to rise the temperature. The volatile organic compounds
(VOCs) are then condensed and collected along with the steam. Hence, volatiliza-
tion technique can be successfully used for the extraction of VOCs from sludges and
wet soils (Fox 1996).
3.2.4 Froth Flotation
Froth flotation is a physio-synthetic method to remove the metal pollutants from

contaminated soils. The setup consists of a chamber for treating the contaminated
soil-water mixture. The reagent mixture is added into the chamber and is dispersed
using air bubble suspension system. The concentrated froth which rises up is col-
lected from the top of the chamber for further treatment and extraction of the con-
taminants (Fig. 3.3). Froth flotation procedure is mainly used in industries for the
extraction of metals from their respective ores. This process has been effectively
used to extract metals such as Pb, Cu, Cd, and Zn, from silts and soils (Cauwenberg
et al. 1998; Venghaus and Werther 1998; Bergeron et al. 2001). The efficiency of the
process depends upon the affinity of a molecule’s hydrophobic surface for air bub-
bles infused in the soil slurry. The process includes three stages: (1) adhesion of the
metal-bearing particles to the air bubbles (2) assembling of the air pockets in the
froth (3) and froth division, which drifts up from the slurry. There are different types
of flotation systems including flotation cells and flotation segments. Metal sulfides
are easy to isolate with this method as compared to oxides and carbonates (Bouchard
2001). In soil remediation, the main considerations which affect the flotation of
metal particles are: (1) heterogeneity of the metal mixes, (2) metal conveyance over
the molecular surface and (3) size of the particles (Cauwenberg et al. 1998;
Vanthuyne et al. 2003). The selective flotation of metal particles from the molecule
of small size (<10 μm) is problematic in most of the mechanical flotation
Fig. 3.3 Process of froth flotation to remove the pollutants from contaminated soil
procedures because of the large surface area of hydrophilic undesirable ore particles
(Kirjavainen 1996; Vanthuyne and Maes 2007). Flotation methods are not very
effective for extraction of heavy and coarse metal particles (Bouchard 2001). The
dissolved air flotation (DAF) system produces very small bubbles and is a promis-
ing technology for flotation of small size metal particles (Vanthuyne et al. 2003).
3.2.5 Thermal Remediation
Thermal remediations are the most commonly used methods which involve the uti-
lization of heat so as to volatilize the contaminants from the complex soil mixture.
This treatment generally occurs in two steps i.e. primary desorption and secondary
incineration (Fig. 3.4). In situ thermal remediation includes five strategies: (1) steam
injection and extraction, (2) electrical resistance heating, (3) radio-frequency heat-
ing, (4) vitrification and conductive heating. Certain operational parameters such as
time of treatment, heating and temperature can be controlled during thermal treat-
ment. These treatments are most commonly used for rapid soil cleanup by enhanc-
ing the mobility of pollutants (hot air/steam injection procedure), by transforming
the pollutants into less toxic residuals (pyrolysis procedure), by separating the pol-
lutants from soil (microwave heating/thermal desorpion procedure), by immobiliz-
ing the pollutants (vitrification procedure) and by destroying the pollutants
(incineration procedure). Arlai et al. (2012), provides a general analysis that the
effects of heat can alter the physical and chemical properties of organic pollutants.
Fig. 3.4 Schematic representation of thermal treatment for remediation of contaminated soil
It was reported that Vapourization is the main phenomenon used for the recovery of
VOCs. Heat-activated persulfate oxidation of groundwater contaminated by herbi-
cide atrazine has been reported by Ji et al. (2015). They predicted transformation
pathways, including dealkylation, alkyl chain oxidation, and dechlorination-
hydroxylation along with underlying mechanisms under increased temperature.
Although thermal treatments hold a promising potential to remediate the con-
taminated sediments and soils but, the high temperature can alter the soil properties
to a certain extent. In spite of its high applicability and efficiency, very diminutive
work has been done in order to expand the feasibility of these treatments on a pilot-
scale. Thermal remediation involves the following methods:
3.2.5.1 Electrical Resistance Heating (ERH)
Electrical resistance heating (ERH) is a procedure which involves the passage of an

electric current via soil moisture, between the electrodes, thereby producing heat.
Hexagonal electrodes are used to supply the electric currents at a particular depth
through six-phase soil heating (Beyke and Fleming 2002). During the heating of
soil, the clay and silt contents gets heated faster as they are more conductive.
Volatilization and steam formation are the two most important removal mechanisms
for removal of many organic and inorganic contaminants (Beyke and Fleming
2005). According to U.S. EPA (2004) near about $32 – $73 per cubic yard are
required for two full-scale ERH for soil remediation. Beyke and Fleming, (2005)
has estimated that approximately $40–$70 per cubic yard are required for soil reme-
diation. In situ ERH has been used for remediation of 1,4-Dioxane and it has been
found that total 99.8% of the contaminant was removed by this method (Oberle
et al. 2015). Truex et al. (2011) demonstrated the combined effect of zero-valent
iron and electrical resistance heating for effective in situ remediation of trichloro-
ethene and it has been found that these techniques efficiently remove the pollutant
from soil. This technique can be used near the people’s locality area, without dis-
turbing the environmental surroundings. Performance or efficiency of ERH may be
enhanced by using it in combination with other soil remediation techniques.
Furthermore, ERH is now proven as an economic and efficient remediation tech-
nique having the capacity to remediate those polluted sites where others techniques
are not applicable (Wolf et al. 2009).
In the year 2003, Department of Energy, US successfully remediated the soil of
Young-Rainey STAR Center, Largo, Florida, contaminated with 2-dichloroethylene,
trichloroethylene, methylene, petroleum hydrocarbons, chloride and toluene.
Remediation was achieved (99.85%–99.99% remediation efficiency) within
4–5 months using the two methods i.e. ERH and steam-enhanced extraction.
Fig. 3.5 Steam injection and extraction process for removal of soil contaminants
3.2.5.2 Steam Injection and Extraction
Steam injection and extraction involves the injection of steam through injection
wells followed by extraction of pollutants, mobilized groundwater and vapors from
the recovery wells (Fig. 3.5). Steam releases its latent heat of vaporization when
injected into the soil thereafter, it condenses into aqueous state and then moves to
soil and finally displaces water and air in front of it. The implementation of steam
injection to a specific contaminated soil site can be determined by the soil permea-
bility, the depth of the pollutants in the soil, and the type of the pollutant. The soil
permeability should be high enough in order to allow sufficient steam to be injected
so as to heat the entire surface (Kempa et al. 2013). Injection pressure should be
increased in order to achieve a higher injection rate. The vaporization of non-volatile
and semi-volatile compounds [n-decane (n-C10), n-dodecane (n-C12),
1,2,4-trimethylbenzene, methylcyclohexane] at temperatures lower than 100 °C
(steam distillation) have been achieved with steam injection procedure (Tzovolou
et al. 2011). Moreover, implementation of this technique is quite difficult as it is a
costly technique and only used in case when high-speed remediation is required.
3.2.5.3 Conductive Heating
Conductive heating involves the process in which heat and vacuum are supplied
concurrently to soil surfaces to degrade the contaminants. Heat can be supplied to
the soil with the aid of thermal wells and thermal blankets. As the soil is heated, the
pollutants present in soil get degraded or vaporized by various phenomena such as
(a) distillation (b) oxidation (c) boiling (d) evaporation and (e) pyrolysis. These
degraded contaminants are then extracted through the vacuum wells and then treated
further.
This method expends huge amount of energy and is applicable below as well as
above 100 °C for removal of different types of pollutants such as pesticides, mer-
cury, dioxins, PAHs, PCBs, chlorinated solvents, explosive materials and heavy
metals. Surface heater blankets can be used when treatment zone is nearly 6 inches
to the ground surface. The wells are created by direct push or by using some con-
ventional drilling strategies. Heat wells are made up of steel pipe and current is
supplied through them. The heating elements usually operate at a temperature range
between 540 and 815 °C (Baker and Heron 2004). Steel pipe and the surrounding
soil are heated by radiant energy and thermal conductance respectively. Generally
conductive heating technique performs best in unsaturated soils but, is also appli-
cable in the case of saturated soils having low hydraulic conductivity. At high tem-
peratures, dry soils including fine grained clay and silt, undergo cracking and
shrinkage which enhances the pollutant extraction (U.S. EPA 2004). This can be
used in combination with steam injection, for treating the sites with complex consti-
tution of sand or gravel. This method has very high remediation capacity and almost
100% cleaning efficiency. Various field experiments have been conducted for soil
remediation by TerraTherm Environmental Services, Inc., USA, by using this
method.
3.2.5.4 Radio-Frequency Heating (RFH)
Radio-frequency heating (RFH) involves in situ heating of polluted soils by apply-

ing alternating electric field of high frequency (Falciglia and Vagliasindi 2015)
(Fig. 3.6). This technique can be applied to dielectric materials. The supply of cur-
rent increases the mobility of polar molecules, which result in generation of
mechanical heat. Radio frequencies of 27.12 MHz, 13.56 MHz, 6.78 MHz, and
40.68 MHz etc. are recommended for scientific, industrial and medical applications.
Application of higher frequencies can enhance the heating rate but, will reduce the
penetration depth. This technique has some unique advantages over microwave
heating, as RFH uses frequencies in MHz range (Roland et al. 2012). Bientinesi
et al. (2015), followed the basic principles of RFH and described its usefulness for
remediation of oil polluted soils, based on several years of work experience at
Consorzio Polo Tecnologico Magona (CPTM), Italy. Their work demonstrated that
RFH can be applied when the utilization of other strategies like steam infusion is
restricted due to logical or topographical hinderances. Though, this method has not
Fig. 3.6 Schematic diagram of radio frequency heating set-up for removal of soil pollutants
been used at a commercial level but, in the coming years this technique will defi-
nitely hold a promising potential for soil remediation.
3.2.5.5 In situ Vitrification (ISV)
In situ vitrification (ISV) involves heating at a temperature of 1400–2000 °C of is a

type of procedure which transforms the polluted soil to crystalline solids and stable
glass (Fig. 3.7). This technology can remove, destroy or immobilize the contami-
nants present in soil or sludge and is most common for the removal of radioactive
wastes and petroleum hydrocarbons (Shearer 1991; Hamby 1996; Fu 2008; Yeung
et al. 2010; Wuana and Okieimen 2011). There are two methods to produce heat and
to melt the polluted soils: (1) first method – involves vitrification of the materials by
using electrodes that are placed in soil (typically 4) and electrical resistance, by
applying high voltage to the electrodes while, (2) second method – involves plasma-
arc technology (ISPV-in situ plasma vitrification process) where the electrodes are
placed in the soil and a high voltage electric current is used to heat the soil to a
temperature between 1600 and 2000 °C (Fox et al. 2001; Bashiri et al. 2015; Dupuis
and Knoepfel 2015).
In situ vitrification process greatly depends upon the presence of alkali metal
oxides in the soil as their higher amount increases the heat-conductivity in the soil.
This is a cost-effective technique with little or no disturbance on site and has been
successfully used for the remediation of oil spillage in the Niger Delta Region,
Nigeria (David and Joel 2013).
Plasma arc in situ vitrification (ISPV) is a new and less used technique which is
based on the principle of plasma arc technology in which electric energy is applied
Fig. 3.7 Schematic representation of in situ vitrification process for removal of soil pollutants
between the two electrodes in the form of direct current. Passage of electric current
at high temperatures (2000–20,000 °C) generates highly ionized gases between the
electrodes and resistance to this electric flow produce plasma. Plasma is considered
as a fourth state of matter which consist of a mixture of ions, electrons and neutral
particles (Jiang et al. 2014). Plasma arc is generated when the voltage applied
between the two electrodes is sufficient to ionize the gas between them and allow
the gas to conduct electric current.
Plasma torch is a device which is used to convert the electrical energy into ther-
mal energy (Camacho 1988, 1991). These torches are operated at extremely high
enthalpy, high temperature range (4000–7000 °C) as compare to fossil fuels burn-
ers. PISV remediation set up involves the introduction of a plasma torch through
drilled borehole into an area of contaminated soil at a particular depth. With an
introduction of this plasma torch, the inorganic fraction of the soil got vitrified while
the organic fraction got pyrolysed. In this way the soil contaminants converted into
highly stable mass which is resistant to leaching (Fox et al. 2001).
Various laboratory based studies have been conducted with ISPV (at 100–200 kW
power) in Georgia Institute of Technology, Atlanta, USA. These studies revealed the
successful remediation of soil contaminated with various organic, inorganic, heavy
metals and radioactive materials by using ISPV (Circeo and Mayne 1993; Mayne
et al. 2000). Plasma arc method has been successfully used to destroy waste rubber
(Huang et al. 2003), to destroy polypropylene (Tang et al. 2005) to destroy (99.99%
efficiency) the polychlorinated biphenyls present in chemical solids (Li et al. 2009)
and hospital wastes (Kołaciński et al. 2017). This method is beneficial over the tra-
ditional and planar in situ vitrification as it is cost effective and highly efficient and
it allows the vapors and gases to escape from the surface of melt zone rather than
being confined to it. This treatment can be used to treat various types of contami-
nants such as toxic materials, organic and inorganic contaminants, wastes sludges
and sediments, underground storage tanks, radioactive wastes, sanitary landfills etc.
Furthermore, the cost of this technology depends upon various factors such as type
of contaminants, conditions of soils and the type of plasma torch (Circeo and Martin
2001).
‘Planar melting’ is an advancement of the in situ vitrification method, where the
starter material is injected in a vertical plane in between the electrodes at a certain
depth. Generally, two electrodes are used with starter material. These electrodes
move away from each other as the melting starts. Planar technique was successfully
performed at the Los Alamos National Laboratory in 2000 (Coel-Roback et al.
2003). This technique has been successfully used to remediate the debris or soil
contaminated with VOCs, SVOCs, PCBs and certain other radioactive materials.
Before the initiation of this technique, complete characterization of waste materials
is a pre-requisite. High applicability, durability and cost effectiveness are the posi-
tive aspects of this technique. Migration of contaminants to other uncontaminated
areas is the limitation of this technique.
Plasma Arc In Situ Vitrification Plasma arc in situ vitrification (ISPV) is a new
and less used technique which is based on the principle of plasma arc technology in
which electric energy is applied between the two electrodes in the form of direct
current (Fig. 3.8). Passage of electric current at high temperatures (2000–20,000 °C)
generates highly ionized gases between the electrodes and resistance to this electric
flow produce plasma. Plasma is considered as a fourth state of matter which consist
of a mixture of ions, electrons and neutral particles (Jiang et al. 2014). Plasma arc is
Fig. 3.8 Schematic representation of in situ plasma vitrification process

generated when the voltage applied between the two electrodes is sufficient to ion-
ize the gas between them and allow the gas to conduct electric current.
Plasma torch is a device which is used to convert the electrical energy into ther-
mal energy (Camacho 1988, 1991). These torches are operated at extremely high
enthalpy, high temperature range (4000–7000 °C) as compare to fossil fuels burn-
ers. PISV remediation set up involves the introduction of a plasma torch through
drilled borehole into an area of contaminated soil at a particular depth. With an
introduction of this plasma torch, the inorganic fraction of the soil got vitrified while
the organic fraction got pyrolysed. In this way the soil contaminants converted into
highly stable mass which is resistant to leaching (Fox et al. 2001).
Various laboratory based studies have been conducted with ISPV (at 100–
200 kW power) in Georgia Institute of Technology, Atlanta, USA. These studies
revealed the successful remediation of soil contaminated with various organic, inor-
ganic, heavy metals and radioactive materials by using ISPV (Circeo and Mayne
1993; Mayne et al. 2000). Plasma arc method has been successfully used to destroy
waste rubber (Huang et al. 2003), to destroy polypropylene (Tang et al. 2005) to
destroy (99.99% efficiency) the polychlorinated biphenyls present in chemical sol-
ids (Li et al. 2009) and hospital wastes (Kołaciński et al. 2017). This method is
beneficial over the traditional and planar in situ vitrification as it is cost effective
and highly efficient and it allows the vapors and gases to escape from the surface of
melt zone rather than being confined to it. This treatment can be used to treat vari-
ous types of contaminants such as toxic materials, organic and inorganic contami-
nants, wastes sludges and sediments, underground storage tanks, radioactive wastes,
sanitary landfills etc. Furthermore, the cost of this technology depends upon various
factors such as type of contaminants, conditions of soils and the type of plasma
torch (Circeo and Martin 2001).
3.3 Conclusions
Physical soil remediation technique includes stepwise removal of contaminant

through washing or volatilization treatment. It is a type of volume reduction process
which further transfers the concentrated pollutant into another media i.e. water and
air. From that media, these contaminants are further treated and converted into more
concentrated form. Depending upon the concentration of the contaminant(s) most
suitable strategy can be designed/selected in order to fully destroy the contaminant(s).
Electro kinetic and soil flushing methods should be completely understood before
their large-scale implementation. Removal efficiency of physical treatments for
various volatile organic compounds can be enhanced with the aid of steam, hot air
or electric current. Physical treatments can remove both heavy metal and organic
contaminants in the same treatment system and the extracted metals can be recycled
in some cases. Moreover, for large scale implementation of physical methods there
References 73
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contaminants should be avoided from the contaminated area as it may cause serious
health related issues. To conclude, merging of different physical treatments along
with the knowledge of plant and soil science is crucial for designing a suitable,
highly efficient and ecofriendly physical remediation strategy.
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Chapter 4
Chemical Methods of Soil Remediation
Abstract Chemical methods aim at addition of chemicals or solvents into the pol-
luted soils so as to stabilize the pollutants and convert them into less toxic forms that
are harmless to the waterbodies, plants, and human beings. Since, complete soil
remediation is difficult to achieve with biological methods alone hence, the amalga-
mation of both biological and chemical methods has gained much attention of the
scientists. Besides that, the harmful effects of the use of chemical methods should
also be considered before implementing on a pilot-scale. The materials generally
used for chemical treatments are metallic oxides, clays or biomaterials. Remediation
efficiency of these materials depends upon the soil texture, organic matter present in
the soil, type of metal contaminant etc. Furthermore, chemical methods can offer a
fast remediation compared to slow bioremediation process. This chapter discusses
the available chemical remediation methods such as chemical leaching, chemical
oxidation, chemical fixation and electro kinetic remediation.
Keywords Chemical leaching · Chemical oxidation · Chemical fixation ·

Electrokinetic remediation
4.1 Introduction
Chemical treatments involve the addition of certain chemicals in order to remove

the pollutants from contaminated soils, sludges and waste water. A number of chem-
ical treatments are available for soil remediation, which include chemical leaching
and fixation, electro kinetic remediation and vitrification. These methods can be
performed on site, are fast and cost-effective. Chemical remediation techniques
include the following methods:

78 4 Chemical Methods of Soil Remediation
4.2 Chemical Leaching
This method involves washing of the contaminated soil with chemical reagents,
gases or other fluids which help to leach the heavy metals (Pd, Zn, Hg etc.) from the
polluted soils (Ou-Yang et al. 2010). These heavy metals are then recovered from
the leachate. Different concentrations of sulfuric acid (H2SO4), nitric acid (HNO3),
hydrogen chloride (HCl), phosphoric acid (H3PO4), and hydrogen fluoride (HF)
were investigated for their effect and efficiency of arsenic (As) removal from yel-
lowish brown contaminated soil (As 2830 mg/kg). Phosphoric acid has been
reported to be most promising for the extraction of As (Tokunaga and Hakuta,
2002). In another report, potassium phosphate (300 mM, pH 6.0 at 40 °C) was
found to be most effective for removal of As from stream sediments around the
abandoned mine areas in Goro, Korea (Lee et al. 2007). The efficiency of the wash-
ing procedure has been analysed with a combination of cyclodextrin and EDTA, for
mobilization of the contaminants (Ehsan et al. 2007). However, EDTA is a potential
secondary pollutant because of its poor degradability. Li et al. (2009) checked the
efficacy of tea saponin for removal of contaminants. The results showed that the
percentage removal of zinc (Zn), cadmium (Cd), lead (Pb), and copper (Cu) were
6.74%, 42.38%, 13.07% and 8.75%, respectively. Tea saponin can effectively
remove heavy metals as well as soluble acid which will greatly reduce ecological
hazards. Chemical leaching was used to remove heavy metals like nickel (Ni) and
vanadium (V), which were present as impurities in the carbon black waste generated
from gasification of crude oil bottom in refineries. Acid leaching with 2M nitric acid
(HNO3) was found to be a very effective method (more effective than alkali) for
removal of both Ni (95%) and V (98%). Further, the acid treated carbon black waste
was used as an absorbent for dye removal. Interestingly, the adsorbtion capacity of
the treated carbon black waste was very high (∼361.2 mg dye/g carbonblack) than
the untreated one (Dong et al. 2017). Xu et al. (2016), suggested chemical leaching
to be an efficient method for extraction of Plutonium (Pu) from soils contaminated
with radioactive waste. Although, chemical leaching is a successful method for effi-
cient and quick remediation of polluted soils, but, its efficiency depends upon the
nature of soil and the type of heavy metal(s) present.
4.3 Chemical Fixation
Chemical fixation involves the addition of chemicals or materials to the polluted

soil, to stabilize the pollutants, converting them into less toxic form and thus reduc-
ing their bioavailability, mobility and ecological risk (Wuana and Okieimen 2011).
The materials which can be used are metallic oxides, clays or biomaterials. Zhou
et al. (2017) evaluated the synergistic effect of two amendment materials, red mud
(R) and compost (C), for remediation of agricultural soil contaminated with heavy
metals like copper (Cu), lead (Pb), zinc (Zn) and cadmium (Cd). The result
4.4 Chemical Oxidation 79
suggested that a combination of these two materials (R + C) could bring a reduction

in the availability of heavy metals to lower their ecological risk. In situ chemical
fixation has been reported to be a cost-effective technique for removal of arsenic
(As) from contaminated soil (Yang et al. 2007). Several chemicals (single and in
combination) like Ferrous sulfate (FeSO4), potassium permanganate (KMnO4) and
calcium carbonate (CaCO3) were tested for their extent of chemical fixation of As
(formation of insoluble arsenic-bearing phase to decrease its leachability). Treatment
solution containing only FeSO4 proved to be the best for remediation of
As-contaminated soil (Yang et al. 2007). In a study, the ability of bentonite, sodium
bentonite, and diatomaceous earth for remediation of soil contaminated with cad-
mium (Cd) was evaluated. The results showed that the percent remediation of Cd
was 21.40, 27.63, 32.30 and 27.24 when the Cd concentration was 20, 30, 40 and
50 g/kg, respectively (Lv et al. 2009). In another study, the chemical fixation poten-
tial of furfural dreg, phosphate rock and weathered coal was evaluated and it was
found that they had the ability to reduce the heavy metals like Cu, Pb, Cd and Zn, to
some extent (Zhang et al. 2009). Zeolitic imidazolate has also been reported as an
efficient catalyst for chemical fixation of CO2 at room temperature
(Kuruppathparambil et al. 2016). However, chemical fixation is basically deployed
to remediate the soils with lower pollutant(s) content. Undobtedly, the bioavailabil-
ity of the treated/fixed contaminants is influenced by the environmental conditions
(Bolan et al. 2003).
4.4 Chemical Oxidation
In situ chemical oxidation is a widely used remediation strategy in which chemical

oxidation of contaminants is performed in order to degrade wide varieties of con-
taminants including heavy metals and hydrocarbons. Other physical and chemical
treatments are usually not recommended for removal of aromatic hydrocarbons, as
high concentration of solvents is required for complete removal (Luthy et al. 1994,
Masten and Davies 1997; Silva et al. 2005). In order to overcome these limitations
chemical oxidation has been proved to be a promising and most effective remedia-
tion strategy for removal of Polycyclic aromatic hydrocarbons (PAHs). Different
types of oxidants including hydrogen, ozone, persulphate, permanganate and perox-
ides have been evaluated for their efficiency in extraction of contaminants from
polluted soils (ITRC 2005; Zhao et al. 2011a, b). Among them activated persulfate
and permanganate has been reported to show maximum PAHs removal capacity
(Zhao et al. 2011a). AOPs i.e. advanced oxidation processes is a modified form of
chemical treatments which include the use of various oxidants in an integrated way.
This approach has been proved highly effective for removal of various recalcitrant
organic compounds. Among AOPs, perozone, Fenton’s reagent and activated per-
sulfate are the most commonly used oxidants. Removal efficiency of oxidation
methods can be increased to a great extent when used in an integrated manner with
various other bioremediation methods. Various researches have concluded that
chemical oxidation successfully increases the biodegradation of various soil pollut-

ants (Nam et al. 2001; Haapea and Tuhkanen 2006; Kulik et al. 2006; Rivas 2006;
Sun and Yan 2008). Higly pressurized hot water (250–300 °C) have also been used
as an oxidant i.e. extracting agent (Dadkhah and Akgerman 2002; Kronholm et al.
2003). This technique is called supercritical water oxidation (SWO). Chemical oxi-
dation processes have various advantages over biological methods: (1) it can be
used in combination with other remediation methods, (2) it requires lesser time
period for extraction of pollutants, (3) the technique can be easily modified by
changing the oxidants or temperature and pressure conditions, and (4) this tech-
nique can be performed under in situ as well as ex situ (Rivas 2006). On the con-
trary, the reactants other than the target contaminant can react with the oxidants
thus, affect their efficiency. Besides that, the by-products of chemical oxidation are
of potential risk to the environmental (Liao et al., 2018). Oxidation with Fenton’s
reagent produces a group of by-products i.e. quinones, carboxylic acids, ketones
and aldehydes (Lee et al. 2001). Oxidation with permanganate, ozone and persulfate
produces aromatic ketones, aliphatic or aromatic acids, short-chain alkanes, mono-
carboxylic acids, aldehydes, aromatic quinones etc. (Liang et al. 2009; Forsey et al.
2010; Liao et al. 2014a, b). Further researches are required in order to achieve maxi-
mum pollutant removal efficiency and to delimit the production of toxic
by-products.
Electro Fenton process can also be used in a combination with soil washing or
soil flushing remediation method so as to separate the contaminants from soil.
Environmental factors such as toxicity and biodegradability of effluents, operating
parameters such as current density, type of electrode and catalyst used and the com-
position of soil matrix are the major factors which can influence the electro fenton
process (Mousset et al. 2017). The combination of soil washing/soil flushing with
electro fenton process has also been used for the removal of organic contaminants
for e.g. polycyclic aromatic hydrocarbons (PAHs) and petroleum hydrocarbons
(Rosales et al. 2009; Mousset et al. 2014a; b, 2017; Huguenot et al. 2015) explo-
sives e.g. trinitrotoluene (Murati et al. 2009), pesticides e.g. pentachlorophenol
(Hanna et al. 2005) and dyes e.g. Lissamine Green B (Rosales et al. 2009) from
polluted soils.
4.5 Electrokinetic Remediation
Electrokinetic remediation strategy is performed by applying an electric current on

both sides of the contaminated soil so as to generate an electric field gradient
(López-Vizcaíno et al. 2017). In the presence of the electric field, contaminants are
carried to the poles through electrophoresis, electroosmotic flow and electromigra-
tion (Fig. 4.1).
This method is applicable for permeable soil and can be easily performed with-
out any environmental risk thus, protecting the natural ecosystem (Virkutyte et al.
2002; Luo et al. 2004; Xu et al. 2006). Direct electrokinetic remediation is not able
4.6 Conclusions 81
Fig. 4.1 Electrokinetic remediation
to maintain the pH of the soil. It requires addition of certain buffer solutions in the
cathode and anode to control the pH value (Gill et al. 2014). This technique is quite
effective for the treatment of both saturated and unsaturated soils. The remediation
efficiency of this method gets hampered by the presence of hematites, carbonates
and gravel, along with the pollutants. There are several electrokinetic remediation
technologies available at commercial level such as, Elektro-Klean™, Lasagna™,
electrobioremediation etc. (Virkutyte et al. 2002). Electrokinetic remediation can be
made more effective when used in combination with other techniques such as ‘per-
meable reactive barrier’ (PRB) (Zhang and Sun, 2007), electrokinetic-microbe
combined remediation (Yu et al. 2009), and electrokinetic-oxidation/reduction com-
bined remediation (Hamdan et al. 2014).
Still, there are various limitations such as electric energy consumption and
adverse effects of electricity on soil characteristics to carry out the soil remediation
with electro kinetic methods (Pazos et al. 2012; Jeon et al. 2015; Li et al. 2016). In
place of electric energy/DC power, solar energy can be deployed for the treatment
of contaminated soil (Yuan et al. 2009; Hassan et al. 2015; Souza et al. 2016).
4.6 Conclusions
As mentioned before, chemical treatments generally involve the applications of

various solutions or fluids containing chemical reagents i.e. salts, acid or bases,
chelators and surfactants in order the transfer the metal from solid to liquid media.
The efficiency of extraction process depends upon the type of reagent or extraction
method used. Chemical treatments are more advantageous as compared to physical
separation methods. In chemical methods, a variety of heavy metal components can

be treated; the adsorbed metal compounds can be extracted; very fine-grained soil
can also be treated; and the extracted metals can further be recovered by various
other methods. Soil washing with nontoxic or degradable chemical reagents will
surely become a most suitable washing method for soil remediation because of their
less hazardous effects. However, there are various limitations of chemical methods
for their large scale implementation, chemical reagents may affect the cost of treat-
ment; by-products/residuals of chemical reagents may prove to be detrimental to the
soil microbes as well the soil composition; extraction of contaminants from highly
polluted sludges is difficult to operate; treated soil cannot be used for re-vegetation
in various cases because of its altered chemical, physical and biological properties;
and the presence of chemical reagents in water is a kind of water pollution.
Furthermore, these chemical reagents can cause other environmental problems
which may disturb the balance of the ecosystem. To conclude, the chemical and
physical treatments should be deployed in an integrated manner in order to compen-
sate for the limitations, to establish a cost-effective, feasible and ecofriendly soil
remediation strategy.
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Chapter 5
Biochar and Soil Remediation
Abstract Biochar is the charred organic matter formed under high heat and low
oxygen conditions that occur in natural fires and modern pyrolysis systems. Biochars
have various properties for the remediation of polluted soils which include negative
charge and large surface area. The use of biochar facilitates nutrient availability,
enhance the microbial activity, soil organic matter availability, water holding and
enhance crop production of soils. Biochar have excellent potential to adsorb the
contaminants from soil solution and make them unavailable to organisms. Various
methods come under biochar technique which includes carbon sequestration, nutri-
ent exchange, water holding, adsorption/absorption and oxidation/reduction.
Although this method (use of biochar in soil remediation) is simple, robust and suit-
able for many regions of the world but, its economic estimations and optimization
should be taken into consideration for its large-scale implementation. Furthermore,
various other public health related concern associated with biochar use should be
addressed properly in order to establish biochar as a best alternative to other soil
remediation methods, in future.
Keywords Carbon sequestration · Nutrient exchange · Water holding · Adsorption
5.1 Introduction
The use of biochar is one of the recent and effective biological process for carbon
sequestration, soil conditioning and remediation, and water remediation (Lehman
et al. 2006, and 2007; Lehman and Joseph 2008; Sohi et al. 2009; Ahmad et al.
2013). According to International Biochar Initiative (IBI), biochar is a solid sub-
stance or material which can be produced by carbonization of biomass (Mulabagala
et al. 2015). The use of biochar promotes several activities: (1) it facilitates the
availability of nutrients in soil, (2) enhances the activity of microbes, (3) enhances
the availability of soil organic matter, and (4) promotes water holding and crop
production by the soils. Thus, its use decreases the fertilizer requirements of the
soil, emission of greenhouse gases, leaching of nutrients and soil erosion (Sohi et al.
2009; Woolf et al. 2010). The metal ions, organic and industrial effluents are the

86 5 Biochar and Soil Remediation
Fig. 5.1 (a) Soil contaminated with heavy metals and PAHs. (b) Contaminated soil with fertilizer.
(c) Contaminated soil with Biochar. (d) Contaminated soil with Biochar and Fertilizers
main source of water and soil contamination. Biochars have an excellent potential
to absorb inorganic and organic contaminants (Hale et al. 2011; Mohan et al. 2012)
thus, possess the capacity for both water and soil remediation. Biochars have
received great attention recently for extraction of contaminants from contaminated
soil, because of their capability to retard the bioavailability of organic and inorganic
pollutants (Cao et al. 2009; Beesley et al. 2011; Gomez-Eyles et al. 2011; Hale et al.
2011). In future, the use of biochar shall replace coal and wood-based activated
carbons for the deduction of pathogens and pollutants from soil (Bernd et al. 2013).
A decrease in the bioavailability of organic contaminants has been observed by
using this technique for agricultural soils (Yang et al. 2006; Cao et al. 2009; Wang
et al. 2010; Yu et al. 2011; Li et al. 2017). In a recent report by Liu et al. (2017),
biochar in combination with immobilized bacterial consortium effectively removed
82.18% of cypermethrin from soil, within 40 days, and the shortest degradation
half-life of cypermethrin was observed to be 16.4 days. Therefore, the use of bio-
char increases soil health and productiveness, and can boost the agricultural yields
(Qambrani et al. 2017). The integrated approach of biochar with phytoremediation
can enhance the growth and remediation efficiency of plants grown on mine tailings
(Fellet et al. 2014). Combination of biochar with composting has been successfully
reported to enhance the remediation efficiency. Composting amends the biochar
5.3 Nutrient Exchange 87
surface structure and biochar stimulates the microbial activity of the compost (Wu
et al. 2016). Use of biochar in combination with soil fertilizers can effectively
reduce the leaching of contaminants into soil and the availability of heavy metals to
the plants. This would enhance the efficiency of fertilizers thereby increasing the
crop yields (Fig. 5.1) (Beesley et al. 2011; Karami et al. 2011; Sizmur et al. 2011;
Beesley et al. 2014). Biochar does not require further refinement as this technique is
simple, robust and appropriate for many areas of the world. But, for its large-scale
application, economic evaluations and optimization are a pre-requisite. Biochar-
mediated remediation involves the following methods.
5.2 Carbon Sequestration
Carbon sequestration is the accumulation or storage of CO2 or other carbon forms

so as to avoid the climate change. Concentrations of pesticides have been reported
to decrease in polluted soil with an increase in the concentration of biochar content
in the soil (Singh and Kookana 2009). The ability of the biochars to remove the pol-
lutants from soil and sediments through sequestration has been successfully evalu-
ated (Lehmann and Joseph 2015; Luo and Gu 2016; Guo et al. 2017). In a report by
Luo and Gu (2016), the addition of biochar, led to a decrease in the microbial abun-
dance and their enzymatic activity but, compared to that of controls, the carbon
sequestration activity was increased (Luo and Gu 2016). In a study by Guo et al.
(2017), multi-walled carbon nanotubes were used as an absorbent for in situ seques-
tration of dichlorodiphenyltrichloroethane (DDTs) and hexachlorocyclohexanes
(HCHs). Carbon nanotubes were found to sequester these pesticides efficiently due
to their enormous adsorption capacity and surface area.
5.3 Nutrient Exchange
Nutrient exchange is a chemical process in which nutrient ions (cations and anions)
get stabilized in soil instead of being leached away. The cation exchange capacity
(CEC) is an indicator of the absorbance capacity of soil to absorb cations. The soils
with high CEC value are usually fertile as they have high nutrient holding capacity.
Anion exchange usually takes place at positively charged places due to the dissocia-
tion of hydroxyl ions from aluminum and iron hydroxides, clay minerals, and
amide/amine groups present in the organic matter of soil (Singh et al. 2017). The
pyrolysis of organic substances and their further exposure to the environment leads
to oxygenation of biochar surfaces. These oxygenized surfaces produce oxygen
containing functional groups including hydroxyl, carboxyl, phenol and carbonyl
groups having high CEC which breakdown the organic contaminants into their
byproducts (Cheng et al. 2006; Liang et al. 2006; Lee et al. 2010). With an increase
in pyrolysis temperatures the CEC of biochars first increases and then decreases
which ultimately broken down the contaminants into small compounds (Gaskin
et al. 2008; Lee et al. 2010; Harvey et al. 2011; Mukherjee et al. 2011).
5.4 Water Holding
Soils supplemented with biochar efficiently increased the nutrient holding capacity
and nutrient composition of soil, due to the direct addition of nutrients (Gaskin et al.
2008; Novak et al. 2012; Brantley et al. 2015). Addition of biochar into the soil is an
effective treatment for removal of toxicants from polluted soils due to its various
properties such as: high-water retention capacity; reduction of bioavailability; capa-
bility to retain pesticides; protection of microbes from predators prevents the stress
induced by toxin to plants and microorganisms. Recently, workers have focused
their attention on evaluating the use and suitability of biochar as a soil remediation
strategy for several benefits such as: (1) to enhance the seed germination, (2) to
enhance the productivity of crop and soil and, (3) to promote vegetation on indus-
trial and sewage waste contaminated soils, waste rock piles and mine tailings
(Anawar et al. 2015). All biochars may not prove beneficial for the degradation of
soil contaminants because moisture content of soil moisture content which will
affect the degradation rates of the respective toxicants.
5.5 Adsorption/Absorption
Black carbons are present in natural conditions by the partial incineration of fossil
fuels and biomass. They can also be synthesized using a pyrolyzer. The adsorption
of the hydrophobic organic pollutant by black carbon-like materials can efficiently
retard their concentration in the aqueous phase (Ghosh 2007; Yang and Xing 2010;
Pignatello 2013; Kołtowski et al. 2016). The applications of black carbon into the
soil decrease the availability of organic pollutants in the soil (Xu et al. 2015). The
activated carbon (black carbon produced from pyrolysis of different materials at
high temperature) can effectively retard the accumulation of pollutants in soil, slud-
ges and sediments (Cho et al. 2009; Beckingham and Ghosh 2011; Ghosh et al.
2011). Biochar produced from pyrolysis of various agricultural crop residues has
been proved to be effective for the adsorption of organic contaminant atrazine and
heavy metal Pb, from the polluted soil (Cao et al. 2009).
5.6 Oxidation/Reduction
The redox reactions i.e. oxidation and reduction reactions in the soil are essential
as they regulate the solubility of many elements, create new compounds and hence,
alter the soil biochemistry. With the addition of biochar these reactions get enhanced
5.7 Biochar-Microbe Interaction in Soil 89
and so it can be used for the remediation of soil contaminants. The application of
biochar increases the soil porosity while decreases the soil density (Mukherjee and
Lal 2013; Ma et al. 2016). These large and small pores containing air and water are
congenial for oxic and anoxic reactions to occur. Arsenic is one of the toxic metals
and is more poisonous in its reduced form As (III) under anoxic conditions, as
compared to As (V) formed under oxic conditions. Reducing conditions may
develop in the small micropores due to increased aeration, which resulted into
decrease in the valence state of As (Ratnaike 2003; Moreno-Jiménez et al. 2012).
Oxides have large affinity for the heavy metals and form hydrogen, covalent or
ionic bond with them (Kunhikrishnan et al. 2016; Nagarajah et al. 2017; Yu et al.
2017). Iron oxide is one of the best-known sink for many heavy metals including
arsenic (As), mercury (Hg), chromium (Cr), lead (Pb), and selenium (Se) etc.
hence, iron oxides can be used for remediation of soil (Pardo et al. 2016; Zou et al.
2016; Sharma et al. 2017). Certain other oxides such as Al and Mn oxide also have
same characteristics. Dissolved organic carbon of biochar has been reported to
reduce and oxidize the arsenic (As) and cadmium (Cd) into their less toxic forms
i.e. As (III) and Cd (VI) (Dong et al. 2014). The effects of different forms of Fe
oxides such as Fe salts, red mud, Fe(0), or iron containing by-products in polluted
soils have been evaluated by supplying it to the contaminated soil and it was found
that Fe oxides enhanced the immobilization of trace elements present in soil (Suda
and Makino 2016; Kim et al. 2017).
5.7 Biochar-Microbe Interaction in Soil
The term ‘biochar’ generally refers to a carbon-rich solid material which is pro-
duced as a result of pyrolysis of biomass (plant-based materials) under anaerobic
conditions (Chen and Chen 2009; Beesley et al. 2011). Because of their prospective
benefit for soil fertility, carbon sequestration and contaminant immobilization bio-
chars are deployed in soil remediation strategies (Cao et al. 2009; Jeffery et al.
2015). A combination of contaminant-degrading microbes and biochar can increase
the degradation of PAHs contaminated soil (Chen et al. 2012; Garcia-Delgado et al.
2015). The property of biochars to facilitate the immobilization of pollutants for
further degradation by the microbes have drawn the attention of soil scientists (Oh
et al. 2013; Dong et al. 2014).
Biochar takes part in the direct extracellular electron transfer (DEET) between
the microbial cells and soil minerals or organic matters and also takes part in direct
interspecific electron transfer (DIET) between the microbial cells (Chen et al. 2014;
Fang et al. 2014). This transfer of electrons between microbial cells and biochar
plays a chief role in the degradation of heavy metals and pollutants present in the
soil (Fang et al. 2015; Yang et al. 2016a).
Although biochar influences (both positive and negative) the microbial activity
in several ways, seven possible mechanisms have been reported (Fig. 5.2): (1) bio-
char facilitates microbial territories by improving soil characteristics such as aera-
tion conditions, pH and water content that are crucial for microbial growth (Quilliam
Fig. 5.2 (1) Biochar provide shelter to microbes in its pores (2) Biochar can improves soil CEC
and enhance microbial growth via sorption of nutrient cations with functional groups (3) VOCs
and free radicals inhibit the growth of toxic microbes and enhance plant growth (4) Biochar
improves soil conditions (e.g. water content, aeration, pH) (5) Biohar improves enzyme activities
by absorbing enzyme molecules (6) Biochar increase rate of hydrolysis of signaling molecules and
change the structures of microbial community (7) Biochar enhance adsorption and degradation of
soil pollutants and thus reduce the toxicity of pollutants
et al. 2013); (2) biochar provides shelters to soil microbes in their pore surfaces and
structures (Quilliam et al. 2013); (3) biochar provides sufficient amount of nutrients
to the soil microbes and thus facilitated their growth (Joseph et al. 2013); (4) bio-
char effects the enzymatic activities of microbial communities (Lehmann et al.
2011; Yang et al. 2016b); (5) due to the hydrolysis of signaling molecules, biochars
disturbs the intra-and inter-specific signaling between the microbial cells (Gao et al.
2016); (6) biochar decreases the bioavailability of pollutants to microorganisms by
absorbing and degrading the pollutants (Qin et al. 2013; Stefaniuk and Oleszczuk
2016); and (7) biochar triggers potential toxicity with hydrocarbons and environ-
mentally persistent free radicals (Fang et al. 2014). In a nut-shell, biochar provides
a microenvironment essential for microbial activity.
5.8 Remediation of Organic and Inorganic Contaminants from Soils 91
5.8 emediation of Organic and Inorganic Contaminants

R
from Soils
The greatest concern of organic contaminants has been focused on herbicides, pes-
ticides, dyes, polycyclic aromatic hydrocarbons and antibiotics (Qiu et al. 2009,
Beesley et al. 2010, Zheng et al. 2010, Teixidó et al. 2011, Xu et al. 2012). The vari-
ous mechanisms proposed for the interaction of biochar with organic and inorganic
contaminants is shown in Figs. 5.3 and 5.4. In a study by Jones et al. 2011, it has
been reported that in soil contaminated with simazine application of biochar signifi-
cantly suppresses the leaching and biodegradation of simazine into the groundwater.
Small particle size and high application rate (25 t/h) of biochar was found to be most
effective for adsorption of simazine. Similarly biochar produced from cottom straw
and woodchips (850 °C) has been reported to significantly decrease the dissipation
of fipronil, chlorpyrifos and carbofuran from contaminated soil and thus reduce
their bioavailability (Yu et al. 2009; Yang et al. 2010).
Metal or other inorganic contaminants are produced from various anthropogenic
sources such as metal finishing, pesticides, mining, animal manure, power plants,
fertilizers, smelting, battery manufacture, sewage sludge, waste water etc. (Adriano
2001; Ok et al. 2011; Usman et al. 2012; Lim et al. 2013). The metals are non-
biodegradable and are highly toxic to all the living being (Adriano 2001; Zhang
Fig. 5.3 Postulated mechanisms of the interactions of biochar with organic contaminants. (a)
Electrostatic interaction between biochar and organic contaminant. (b) electrostatic attraction
between biochar and polar organic contaminant. (c) Electrostatic attraction between biochar and
non-polar organic contaminant
Fig. 5.4 Postulated mechanisms of biochar interactions with inorganic contaminants. (a) Ion
exchange between exchangeable metal and metal contaminant in biochar, (b) electrostatic attrac-
tion of anionic metals (c) precipitation of target metal contaminants and (d) electrostatic attraction
of cationic metal
et al. 2013). Biochar is a novel pyrolysed material which can absorb or adsorb these
metals from polluted soil or water. Table 5.1 summarizes the various studies on
biochar applications for remediating soil contaminated with organic and inorganic
contaminants. In a study by Beesley et al. (2010), they have used the biochar
obtained from hardwood in the soil contaminated with Cd, Zn, Cu and As. It has
been reported that among these metals Cd and Zn immobilized in soils amended
with biochar as compared to un-amended soil. Mobility of As in the soil was also
reported to increase with the addition of biochar (Park et al. 2011b; Zhang et al.
2013). In a similar report, mobility of antimony also increased with the addition of
biochar derived from broiler litter (Uchimiya et al. 2012).
Overall, the biochars produced at higher temperatures exhibit higher sorption
efficiency for organic contaminant remediation in soil and water. This is probably
due to the high surface area and microporosity of biochars. Therefore, the biochars
should be produced under well-defined pyrolysis conditions. The biochar properties
should also be carefully examined before the applications for the remediation of
specific organic contaminants in soil or water.
5.8 Remediation of Organic and Inorganic Contaminants from Soils 93
Table 5.1 Applications of biochar for remediation of soils contaminated with organic and
inorganic contaminants
Biochar type Contaminant(s) Result Reference
Hard wood Arsenic Mobilization of arsenic due to Hartley
(400 °C) enhanced pH et al.
(2009)
Woodchips (450 Chloropyrifos and Adsorption of contaminants due to Yu et al.
and 850 °C) carbofuran high surface area and nano-porosity (2009)
Pine wood (350 Phenanthrene Entrapment in micro- or meso-pores Zhang
and 700 °C) et al.
(2010)
Hard wood Polycyclic aromatic Sorption and biodegradation Beesley
hydrocarbons (PAHs) et al.
Hard wood Cadmium and zinc Immobilization due to enhanced pH (2010)
Hard wood Arsenic and copper Mobilization due to enhanced pH
Dairy manure Atrazine Sorption Cao et al.
(450 °C) (2011)
Hardwood (450 Simazine Sorption due to abundance of Jones et al.
and 600 °C) micropores (2011)
Rice straw Pentachlorophenol Adsorption due to high surface area Lou et al.
and microporosity (2011)
Chicken manure Cadmium, lead and Immobilization due to partitioning of Park et al.
and green waste copper metals from the exchangeable phase to (2011a)
(550 °C) less bioavailable organic-bound
fraction
Broiler litter Copper Cation exchange; electrostatic Uchimiya
(700 °C) interaction; sorption on mineral ash et al.
contents; complexation by surface (2011c)
functional groups
Oak wood Copper and lead Complexation with phosphorous and Karami
organic matter et al.
(2011)
Dairy manure Lead Immobilization by Cao et al.
(450 °C) hydroxypyromorphite formation (2011)
Cottonseed Nickel, copper, lead and Surface functional groups of biochars Uchimiya
hulls cadmium controlled metal sequestration et al.
(200–800 °C) (2011b)
Pulpgrade Tylosin Sorption Jeong et al.
hardwood and (2012)
softwood chips
(850 and
900 °C)
Oak wood Lead Immobilization by rise in soil pH and Ahmad
(400 °C) adsorption onto biochar et al.
(2012)
Rice straw Lead Non-electrostatic adsorption Jiang et al.
(2012)
(continued)

Biochar type Contaminant(s) Result Reference
Broiler litter Lead, zinc, copper and Stabilization of Pb and cu; desorption Uchimiya
(350 and antimony of Sb et al.
600 °C) (2012)
Sewage sludge Arsenic, cobalt, Immobilization of arsenic, chromium, Khan et al.
(500–550 °C) cadmium, nickel, cobalt, nickel and lead due to rise in (2013)
chromium, copper, lead soil pH; mobilization of copper, zinc
and zinc and cadmium due to high available
concentrations in biochar
5.9 Conclusions
Since biochar has been reported to be beneficial for crop productivity, for soil reme-
diation and reuse of agricultural wastes hence, researches in the field of bioremedia-
tion with biochar have increased rapidly in the past few years. Because of their
multiple applications biochar can be used as an alternative to other bioremediation
techniques. However, we cannot ignore the various public health issues related to
the use of biochar, as the soil amended with biochar can cause several respiratory
diseases. Furthermore, biochar can accumulate high concentration of heavy metal
ions which is also one of the serious health related concern. Hence, the appropriate
dose of biochar should be taken into consideration for its use in the soil remediation
program. Moreover, leaching of the contaminants to ground water from biochar
amended soils and their accumulation in soil microbes will eventually lead to the
contamination of the ground water and bio-magnification of the contaminants
respectively. In this way these contaminants may enter into food-chain or food-web
and thus disturbing the ecosystem. To overcome the aforementioned drawbacks,
further in-depth research is required for establishment of biochar as an ecofriendly
and efficient soil remediation treatment.
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Chapter 6
Soil Remediation Through Microbes
Abstract The use of microbes including bacteria and fungi for treatment of pol-
luted soils is also a method of soil remediation. Microbes are adapted to survive
under various unfavorable conditions, such as high temperature, alkalinity, and
acidity, and can easily develop biological resistance against the toxic substances
because of their jumping genes. Under favorable conditions of growth and adequate
supply of the nutrients, the microbes can biotransform or biodegrade complex
organic materials into harmless or less toxic smaller molecules. With the addition of
micronutrients into the microbial consortia the degradation rate of pollutants can be
enhanced as the added nutrients stimulate soil microbes which eventually expedite
the rate of biodegradation. Mycoremediation is fungal-mediated bioremediation of
polluted soils containing organic and inorganic pollutants. Fungal mycelia can
spread similarly as that of plant roots and accumulate heavy metals in their cytosol.
Environmentalists regard microorganisms as ‘eco-friendly nano-factories’ for treat-
ing the polluted soils. However, natural pollutants can be degraded by the naturally
occurring microbes while for degradation of manmade chemicals or pollutants,
genetically transformed microbes should be developed for efficient soil remedia-
tion. Microbe-mediated remediation can take place via three methods i.e. biovent-
ing, bioleaching, bioaugmentation. In the last few years, several reports covering the
success stories of microbe-mediated soil remediation have come up.
Keywords Microbial remediation · Bioventing · Bioleaching · Bioreactors ·

Bioaugmentation
6.1 Introduction
Microbes play a vital role to sustain the environment (Widdel and Rabus 2001;
Varjani and Srivastava 2015) and have been successfully used for the remediation of
polluted soils (Chandra et al. 2013; Varjani and Upasani 2016). They are adapted to
survive under unfavorable conditions such as high temperature, alkalinity, and acid-
ity. Microbes can easily develop biological resistance against the toxic substances
because of their jumping genes. Under favorable conditions of growth and adequate

102 6 Soil Remediation Through Microbes
supply of the nutrients, the microbes can bio transform or biodegrade complex
organic materials into smaller molecules (de-Bashan et al. 2012). Environmentalists
regard microorganisms as ‘eco-friendly nano-factories’ for treating the polluted
soils (Potysz et al. 2016; Verma and Jaiswal 2016; Chen et al. 2016, 2017).
Microbes for e.g. yeast, bacteria and fungi have the potential to degrade the
hydrocarbons present in contaminated soils (McDonald et al. 2006; Varjani et al.
2015; Varjani and Upasani 2016). The biodegradation efficiency of bacteria has
been reported by several research groups which ranges from 0.003% (Hollaway
et al. 1980) to 100% (Phillips and Stewart 1974), 6% (Jones et al. 1970) to 82%
(Pinholt et al. 1979) for soil fungi and 0.003% (Hollaway et al. 1980) to 100%
(Phillips and Stewart 1974) for marine bacteria.
Nowadays, applications of genetically modified/engineered microbes have
gained great attention of the scientists across different fields so as to enhance the
degradation rate of the toxic compounds. Though modified microorganisms have
been successfully used but, their capability is affected by their competitive potential
towards other microorganisms, indigenous predators and certain other abiotic fac-
tors. Growth inhibition and loss of degradation potential of microbes can hinder the
performance of bioremediation. To overcome these problems, repeated inoculation
of microbes into the soil is recommended. To improve the performance of bioaug-
mentation, a combination of various conventional and molecular methods can also
be used (Cycoń et al. 2017). However, various regulatory environmental and eco-
logical concerns related to genetically modified organisms are the major constrains
for their large-scale implementation.
6.2 Microbial Degradation of Petroleum Hydrocarbons
Degradation of organic pollutants by microbes is one of the most important and

promising phenomenon for cleaning the soils contaminated with petroleum hydro-
carbons (Atlas 1981; Amund and Nwokoye 1993; Lal and Khanna 1996) (Table 6.1).
Petroleum hydrocarbons can be categorized into four groups: phenols, esters,
porphyrins, and ketones (Colwell et al. 1977). Biodegradation of petroleum hydro-
carbons largely depends upon the nature as well as the amount of hydrocarbons
present. Floodgate (1984), repoted 25 genera of hydrocarbon degrading fungi and
bacteria. Several success stories of soil remediation through microbes are available
(Farhadian et al. 2008; Sajna et al. 2015; Varjani et al. 2015; Varjani and Upasani
2016) (Table 6.2). Furthermore, various researches have also been conducted to out-
line the pathway adopted by microbes for degradation of contaminants (Watkinson
and Morgan 1990; Rojo 2009; Jaekel et al. 2013; Ron and Rosenberg 2014; Abbasian
et al. 2015; Meckenstock et al. 2016; Wilkes et al. 2016). The energy produced dur-
ing the degradation of soil pollutants can be used by the microbes for their cellular
metabolism (Abbasian et al. 2015).
6.2 Microbial Degradation of Petroleum Hydrocarbons 103
Table 6.1 List of selected microorganisms which aid phytoextraction of heavy metals
Microbes responsible for
Element Source remediation Reference(s)
Chromium Tanning, paints, pigments, Bacillus pumilus S28, Bacillus Abou-Shanab
(Cr) fungicide subtilis S3 Brevibacterium et al. (2008)
halotolerans S29 Pseudomonas
pseudoalcaligenes S22
Rahnella aquatilis Kumar et al.
(2009)
Bacillus sp. Dhal et al.
(2010)
Desulfovibrio desulfuricans Ock Joo et al.
(2015)
Nickel (Ni) Electroplating, mining and Microbacterium Abou-Shanab
smelting arabinogalactanolyticum et al. (2006)
AY509225 Microbacterium
oxydans AY509223
Bacillus subtilis SJ-101 Zaidi et al.
(2006)
Pseudomonas sp. Kuffner et al.
(2008)
Pseudomonas jessenii PjM15 Rajkumar et al.
(2013)
Bacillus cereus SRA10, Ma et al.
Psychrobacter sp. SRA1 (2009a)
Psychrobacter sp. SRA2
Bacillus cereus SRA10, Ma et al.
Psychrobacter sp. SRA1 and (2009b)
SRA2
Bacillus sp. SN9, Pseudomonas Ma et al.
sp. SRI2, Psychrobacter sp. (2009c)
SRS8
Achromobacter xylosoxidans Ma et al.
Ax10 (2009b)
Pseudomonas sp. A3R3 Ma et al. (2011)
Pseudomonas sp. SRI2, Ma et al. (2011)
Psychrobacter sp. SRS8 and
Bacillus sp. SN9
Bacillus pumilus NBRFT9 Tiwari et al.
Paenibacillus macerans (2012)
NBRFT5
Bacillus endophyticus NBRFT4
Bacillus megaterium SR28C Rajkumar et al.
(2013)
Enterobacter cloacae Banerjee et al.
(2015)
(2015)
(continued)

Lead (Pb) Bearing metals, cable Bacillus firmus Salehizadeh and
covering, explosives, batteries, Shojaosadati
manufacture of pesticides, (2003)
antifouling paints, analytical
reagents and gasoline additives
Vibrio harveyi Mire et al.
(2004)
Corynebacterium glutamicum Choi and Yun
(2004)
Bacillus sp. Tunali et al.
(2006)
Bacillus sp. ATS-2 Cabuk et al.
(2006)
Pseudomonas putida Uslu and Tanyol
(2006)
Bacillus megaterium HKP-1, Wu et al.
Azotobacter chroococcum (2006a)
HKN-5 and Bacillus
mucilaginosun HKK-1
Microbacterium sp. G16 and Sheng et al.
Pseudomonas fluorescens G10 (2008)
Cupriavidus taiwanensis Chen et al.
(2008b)
Burkholderia sp. J62 Jiang et al.
(2008)
Bacillus sp. RJ16 He et al. (2009)
Azotobactor chroococum and Hadi and Bano
Rhizobium leguminosarum (2010)
Rahnella sp. JN6 He et al. (2013)
Enterobacter sp. JYX7and Jing et al.
Klebsiella sp. JYX10 (2014)
Pseudomonas aeruginosa Ahmady-
Asbchin et al.
(2015)
(2015)
Manganese Sewage sludge, Municipal –
(Mn) wastewater discharges, mining
and mineral processing, iron
and steel foundries, emissions
from steel, alloy and iron
production fossil fuels
combustion, industrial
emissions power plants
(continued)

Silver (Ag) Manufacture and disposal of –
certain photographic and
electrical supplies, emissions
from smelting operations, coal
combustion, and cloud seeding
Cadmium Fertilizer, plastic, pigment Arthrobacter mysorens, Belimov et al.
(Cd) Agrobacterium radiobacter, (2004)
Flavobacterium sp. L30,
Azospirillum lipoferum,
Bacillus megaterium HKP-1, Wu et al.
HKN-5, and Bacillus
mucilaginosun HKK-1
Burkholderia cepacia Li et al. (2007)
Pseudomonas tolaasii ACC23, Dell’Amico
Pseudomonas fluorescens et al. (2008)
ACC9, Mycobacterium sp.
ACC14,
Bacillus sp. J119 Sheng et al.
(2008)
(2013)
Burkholderia sp. J62 Jiang et al.
(2008)
(2008b)
Pseudomonas aeruginosa Ganesan (2008)
MKRh3
Bacillus sp. RJ16, He et al. (2009)
Pseudomonas sp. RJ10
Streptomyces tendae F4 Dimpka et al.
(2009)
Paecilo myceslilacinus NH1 Gao et al.
(2010)
Bacillus mycoides, Micrococcus Malekzadeh
roseus et al. (2012)
Fusarium oxysporum Zhang et al.
(2012)
Phyllobacterium Ma et al. (2013)
myrsinacearum RC6b
Klebsiella sp. BAM1 Prapagdee et al.
(2013)
Pseudomonas putida 710, Rani et al.
Comamonas aquatica 710B (2013)
(continued)

Rahnella sp. Yuan et al.
(2013)
Enterobacter intermedius Płociniczak
MH8b et al. (2013)
Micrococcus sp. MU1 and Prapagdee et al.
Klebsiella sp. BAM1 (2013)
Pseudomonas aeruginosa, Liang et al.
Bacillus subtilis, B. cereus, (2014)
B.megaterium
Pseudomonas sp. LK9 Chen et al.
(2014)
(2015)
Bacillus pumilus E2S2 and Ma et al. (2015)
Bacillus sp. E1S2
Cedeceadavisae LCR1 Liu et al. (2015)
Rhodococcus erythropolis
NSX2
(2015)
Serratia sp. RSC-14 Khan et al.
(2015)
Zinc (Zn) Fertilizer Streptomyces rimosus Mameri et al.
(1999)
Aphanothece halophytica Incharoensakdi
and Kitjaharn
(2002)
Thiobacillus ferrooxidans Liu et al. (2004)
Azotobacter chroococcum Wu et al.
HKN-5, Bacillus megaterium (2006a)
HKP-1 and Bacillus
mucilaginosun HKK-1
Burkholderia cepacia Li et al. (2007)
(2013)
Enterobacter sp. NBRI K28 Kumar et al.
(2008)
Pseudomonas sp. PsM6, Rajkumar et al.
P. Jessenii PjM15 (2013)
Bacilus subtilis, B. cereus, He et al. (2010)
Flavobacterium sp. and
Pseudomonas aeroginosa
(continued)

NBRFT5
(2012)
Phyllobacterium Ma et al. (2013)
myrsinacearum RC6b
Enterobacter intermedius Płociniczak
MH8b et al. (2013)
Pseudomonas veronii Long et al.
(2013)
Acinetobacter sp. Tabaraki et al.
(2013)
(2014)
Rhizobium leguminozarum Adediran et al.
(2015)
Bacillus pumilus E2S2 and Ma et al. (2015)
Bacillus sp. E1S2
Arsenic Smelting, mining, and Methanobacterium bryantii McBride and
(As) coal-fired power plants Wolfe (1971)
Enterobacter sp., Aeromonas Cullen (1989)
sp., Nocardia sp.
Achromobacter sp.
Pseudomonas arsenitoxidans Santini et al.
NT-26 (2000)
Clostridium collagenovorans, Michalke et al.
Desulfovibrio vulgaris and D. (2000)
gigas, Methanobacterium
formicicum,
Thermus aquaticus and Gihring et al.
Thermus thermophilus (2001)
Enterobactor cloacae CAL2 Nie et al. (2002)
Sulfurospirillum species NP4 MacRae et al.
(2007)
Staphylococcus arlettae Srivastava et al.
(2013)
Bacillus thuringiensis GDB-1 Babu et al.
(2013)
(continued)

Copper Copper plating, copper Pseudomonas cepacia Savvaidis et al.
(Cu) sheathing, fungicides in (2003)
agriculture Psuedomonas aspleni AC Reed and Glick
(Canola) (2005)
Sphaerotilus natans Beolchini et al.
(2006)
Bacillus megaterium HKP-1 Wu et al.
HKN-5,
Bacillus mucilaginosun HKK-1
(2008a, b)
Achromobacter xylosoxidans Ma et al.
Ax10 (2009a)
Enterobacter sp. Bestawy et al.
(2010)
Acidithiobacillus caldus, A. Kaksonen et al.
ferrooxidans, A. thiooxidans, (2011)
Leptospirillum ferrooxidans
and Sulfobacillus
thermotolerans
Kocuria flava Achal et al.
(2011)
Pantoea agglomerans Jp3-3 Zhang et al.
and Pseudomonas (2011)
thivervalensis Y1-3-9
NBRFT5
(2012)
Bacillus thuringiensis GDB-1 Babu et al.
(2013)
(2014)
Enterobacter sp. K3-2 Li et al. (2016)
Acidithiobacillus thiooxidans Potysz et al.
(2016)
Iron (Fe) Metallurgical, iron and steel Pseudomonas aeruginosa Akintunde et al.
industry, sewage, dust from (2015)
iron mining, fertilizer and
herbicide
(continued)

Mercury Municipal waste combustors, –
(Hg) Medical waste incinerators,
Coal-fired power generation,
manufacture of alkali, metals,
and cement
Cobalt Seawater, volcanic Eruptions, –
(Co) forest fires, exhausts from
burning coal and oil, and from
industrial processes
Selenium Photocopying, photocells, –
(Se) light meters and solar cells
Antimony Mines and industrial facilities, –
(Sb) fumes from burning of oil
fuels (vehicle exhaust gases)
and industrial dust Waste
incinerators, metal processing
works, burning coal,
Bismuth Copper, lead gold and silver –
(Bi) smelting, waste water and
sewage sludge,
pharmaceuticals, cosmetics,
pearlescent pigments, ceramic
glazes, permanent magnets
Various fungal species such as Amorphoteca, Neosartoria, Aspergillus,

Paecilomyces, Talaromyces, Fusarium, Sporobolomyces, Graphium and Penicillium
and yeast species such as Pichia, Rhodotorula, Candida, Yarrowia and Pseudozyma
have been successfully used for treatment of petroleum hydrocarbons from polluted
soils (Atlas 1981; Leahy and Colwell 1990; Boonchan et al. 2000; Sajna et al. 2015;
Wilkes et al. 2016). Bacterial species such as Arthrobacter, Azoarcus, Micrococcus,
Achromobacter, Cellulomonas, Flavobacterium, Brevibacterium, Pseudomonas,
Corynebacterium, Nocardia, Acinetobacter, Ochrobactrum, Stenotrophomaonas,
Vibrio and Marinobacter have been reported to degrade hydrocarbons (Widdel and
Rabus 2001; Roy et al. 2002; Foght 2008; Mittal and Singh 2009; Chandra et al.
2013; Meckenstock et al. 2016; Varjani et al. 2015; Varjani and Upasani 2016).
Biodegradation of contaminants involve catalytic action of various enzymes via
three possible ways: (1) chemotrophic aerobic (2) chemotrophic anaerobic and (3)
phototrophic (Abbasian et al. 2015). Reaction such as hydroxylation, dehydrogena-
tion, oxidation and reduction are responsible for both aerobic and anaerobic degra-
dation pathways (Abbasian et al. 2015; Wilkes et al. 2016). ω-oxidation, β-oxidation,
terminal oxidation and sub-terminal oxidation are the major pathways which are
responsible for degradation of pollutants (Salleh et al. 2003; Abbasian et al. 2015).
Table 6.2 Reports on microorganism-mediated remediation of hydrocarbon polluted soils

Compound(s) Sources Microbe(s) Reference(s)
4-Chloronitrobenzene Rubbers, dyes, plastics, Pseudomonas putida Xiao et al.
soaps, detergents, ZWL73 (2006), Zhen
lubricating oils, et al. (2006) and
agrochemicals, Zhao et al.
pesticides and (2009)
pharmaceuticals
industries
Fenitrothion (O, Insecticide Burkholderia sp. FDS-1Zhang et al.
O-dimethyl O-p-nitro-m- (2006) and
tolyl phosphorothioate) Hong et al.
(2007)
para-Nitrophenol Rubber, dyes, paint, Moraxella sp. Spain and
drugs and fungicides Gibson (1991)
industries Arthrobacter sp. Jain et al.
(1994)
Bacillus sphaericus Kadiyala and
JS905 Spain (1998)
Arthrobacter Labana et al.
protophormiae RKJ100 (2005)
Pseudomonas sp. WBC-3 Liu et al. (2005)
Pseudomonas sp. Zhang et al.
(2009)
Methyl parathion (O, Agricultural and Pseudomonas sp. WBC-3 Liu et al. (2005)
O-dimethyl domestic pesticides,
Op-Nitrophenol fungicides, herbicides
phosphorothioate)
4-Chlorophenol Disinfectants, Arthrobacter Jernberg and
pesticides and chlorophenolicus A6L Jansson (2002)
herbicides
Pentachlorophenol Agricultural and Sphingobium Cai and Xun
industrial uses chlorophenolicum ATCC (2002), Dams
39723 et al. (2007)
4-Chloronitrobenzene Industrial applications Pseudomonas putida Xiao et al.
ZWL73 (2006) and
Zhen et al.
(2006)
Comamonas sp. CNB-1 Wu et al.
(2006b) and Ma
et al. (2007)
Ortho-nitrophenol and Rubber, dyes, paint, Pseudomonas sp. Chi et al. (2013)
metanitrophenol drugs and fungicides WBC-3, Cupriavidus
industries necator JMP134,
Alcaligenes sp. NyZ215
(continued)
6.3 Bioventing 111

Compound(s) Sources Microbe(s) Reference(s)
PAHs mixture Industrial effluents, Rhodococcus sp., Yu et al. (2005)
(phenanthrene, fluorene, diesel and gasoline Acinetobacter sp. and
pyrene) exhaust, soot, coke, Pseudomonas sp.
cigarette and cigar Bacillus sp. B3G, Silva et al.
smoke Bacillus sp. B1F, (2009a, b)
Bacillus sp. B5A,
Enterobacter
agglomerans B1A,
Chromobacterium sp.
4015
Pseudomonas aeruginosa Varjani and
NCIM 5514 Upasani (2016)
Fig. 6.1 Schematic representation of bioventing for treatment of contaminated soil
6.3 Bioventing
The soil bioventing involves the supply of oxygen to the polluted soils to stimulate
microbial degradation of the pollutants. A bioeventing setup consists of a blower or
compressor associated with one air-supply well and soil-gas observing wells
(Downey et al. 2004) (Fig. 6.1). However, this technique is a choice for remediating
light-spilled petroleum products but, may be utilized to treat more recalcitrant tox-
ins using ozonation (Philp and Atlas 2005; Azubuike et al. 2016). Obviously, bio-
venting has a future and a minimal innovation, along with people’s interest for more
natural strategies for remediation will most likely empower the continuing experi-
mentation (Herndon et al. 2013). Bioventing is more viable as compared to land
farming (Sharma 2012). Researchers have recommended that anaerobic conditions
may help in degrading the contaminants like chlorinated solvents and polycyclic
aromatic hydrocarbons (PAHs) thus, opening potential outcomes of venting sub-
strates other than oxygen, for growth of microbes (Antizar-Ladislao 2010).
Besides microbial action, the degradation process can stimulate the mobilization
of other toxic metals like arsenic (As) or manganese (Mn) (Hellekson 1999). In
specific cases, one contaminant may really synergise the degradation of another
contaminant. Chlorinated compounds generally degrade anaerobically, yet some of
them trichloroethylene (TCE) and perchloroethylene (PCE) can be degraded aerobi-
cally in the presence of co-metabolites, which include aromatic compounds like
toluene (Bradley 2003). Considering the complexity of polluted sites bioventing
may be applied with certain other strategies like soil-vapour extraction and air-
sparging. The enthusiasm for bioventing might proceed however, care need to be
taken especially while working with chlorinated solvents and PAHs. The break-
down of both of these compounds may generate other secondary toxins. The limita-
tion of this technique is that it is not applicable when the ground water level goes
several feet down. Low temperature and low permeability also hampers the effec-
tiveness of bioventing (Uddin et al. 2017).
6.4 Bioleaching
Bioleaching has been widely used to dissolve heavy metals such as copper (Cu),
nickel (Ni), zinc (Zn), Aurum (Au), lead (Pb), cobalt (Co), arsenic (As) and others
with the use of bacteria, instead of chemicals. This phenomenon is based on the
unique characteristic of certain bacteria (which can eat the metal content) which
uses two different pathways for dissolution of metal sulfides that are polysulfide and
thiosulphate pathway. The dissolution of metals can be achieved by proton attack
and oxidation reactions (Fu et al. 2010; Vera et al. 2013). Rashidi et al. (2012), pro-
posed a mathematical model for bioleaching of uranium by iron-oxidizing bacteria
Acidithiobacillus ferrooxidans, Thiobacillus ferrooxidans, Leptospirillum ferrooxi-
dans and acidophilic iron-oxidizing bacteria for bioleaching of sulfide minerals
(Romo et al. 2013). Extraction of Cu from mine wastes is one of the commercial
applications of this technique (Watling 2014). This technique can be used in combi-
nation with electrokinetic remediation to dissolve and recycle the heavy metals such
as Cu and Zn (Peng et al. 2011). It can also be used in combination with AOPs
(advanced oxidation processes: Fenton or Fenton like reactions) to enhance the pro-
cess of heavy metal removal from sludge. In Fenton like reactions, hydoxyl (OH)
group is the main reactant which can degrade the waste water sludge (Pham et al.
2010; Lin et al. 2015). Various biosurfactants produced by some microbes such as
Bacillus subtilis, B. pumilus, Brevibacillus parabrevis and Pseudomonas aeruginosa
have been reported to remove phenanthrene (Reddy et al. 2010; Kumar et al. 2012).
6.5 Bioaugmentation 113
Fungal species such as Aspergillus niger, Penicillium chrysogenum, P. simplicissi-

mum, P. purpurogenum can also be used in bioleaching (Rezza et al. 2001). It has
been reported that A. niger could effectively remove 82.1 % Pb, 74.1 % Cu, 87.8 %
Cd and 87.6 % Zn from sewage sludge (Xu and Feng 2016). The limitation of this
technique is that it requires a treatment of atleast 16–20 days, which does affect the
maintenance cost and is therefore not suitable for large-scale application.
6.5 Bioaugmentation
Bioaugmentation is the addition of microbes (pollutant degraders cultured in labo-

ratory) into the soils which harbors a low population of indigenous pollutant degrad-
ers (Castiglione et al. 2016). Indigenous microbial populations do not have the
capability to degrade the extensive variety of contaminants present in mixtures such
as petroleum (Isaac et al. 2016). For remediation of crude oil contaminated soils,
indigenous populations can be supplemented with oil-degrading microorganisms.
Degradation rate can be facilitated by the addition of certain aerobic and anaerobic
microbes (Gram +ve/−ve bacteria and fungi) (Kuppusamy et al. 2016). Different
species of microbes have different ability for oil degradation due to their unique
enzymatic activities. Some microorganisms have the capacity to degrade cyclic,
branched or linear alkanes while, some are able to degrade mono or poly-nuclear
aromatics, and some can collectively degrade both aromatics and alkanes.
Bioaugmentation has been successfully performed for the treatment of various
industrial, agricultural and domestic soils. In the past few years, bioaugmentation
has also been practiced for treatment of soils contaminated with aromatic hydrocar-
bons such as nitro phenols, chlorophenols, nitrobenzene, chlorobenzene, organo
pesticides etc. (Chi et al. 2013; Wang et al. 2014). Treatment of soil contaminated
with nitrobenzene has been efficiently achieved with microbial strain Pseudomonas
putida ZWL73 (Zhao et al. 2009) at laboratory scale. In a study by Hong et al. 2007;
different concentrations (1–50 mg/kg) of fenitrothion (toxic nitrophenolic pesti-
cide) was added into the soil and it was reported that Burkholderia sp. FDS-1 effec-
tively retard the concentrations of fenitrothion in the soil and thus indicating a
promising remediation potential. Other bacterial strains such as Arthrobacter proto-
phormiae RKJ100 have been reported to degrade the para-nitrophenolic compounds
present in soil (Labana et al. 2005). Pseudomonas sp. strain WBC-3 significantly
utilized organophosphate pesticides such as methyl parathion for their cellular
metabolism (Liu et al. 2005; Zhang et al. 2009). Soil contaminated with
4-Chlorophenol (180 μg/g of soil) was supplied with bacterial strains Arthrobacter
chlorophenolicus A6L and it was reported that the inoculated strain efficiently
removed 4-Chlorophenol from 180 to 3 μg/g of soil (Jernberg and Jansson 2002)
Bioaugmentation can be performed by using consortia of both bacterial and fungal
strains. In a study by Silva et al. 2009a, b, a consortium of filamentous fungi
(Aspergillus sp., Achremonium sp., Verticillium sp.), consortia of bacterial sp.
(Bacillus sp. B3G, Bacillus sp. B1F, Enterobacter agglomerans B1A, Bacillus sp.
B5A, Chromobacterium sp. 4015) and a mixture of both bacterial and fungal con-
sortia were used for the treatment of soils contaminated with PAH. Among these
consortia, the combinations of both bacterial and fungal strains have been reported
to be highly efficient for PAHs degradation. Efficiency of bioremediation depends
upon the catabolic activity of microbes as well as their survival in the polluted soils.
It has also been reported that certain indigenous microbial communities resist the
invasion of foreign microbes; hence a suitable coexistence must be present between
indigenous and foreign microbial communities for successful implementation of
bioaugmentation.
Furthermore, detailed study of microbes used for remediation purposes may
prove beneficial for the selection of microbes having high degradation potential.
Well adapted microbial species should be taken into consideration for the successful
bioaugmentation process. The various factors which regulate bioaugmentation
include physical environment, concentration and chemical structure of the pollut-
ants, pollutant availability, nature and size of microbes (Adams et al. 2015).
6.6 Mycoremediation
Mycoremediation is a bioremediation process which involves the use of fungi to

reduce the pollutants present in contaminated water, soils and sludges (organic and
inorganic) (Fig. 6.2) (Table 6.3). Various saprophytic, mitosporic, wood decaying
and ectomycorrhizal fungi with efficient PAHs degradation potential have been
described for their remediation potential. (Bamforth and Singleton 2005; Eibes
et al. 2006; Silva et al. 2009a, b). Fungal mycelia can spread similarly as that of
plant roots and accumulate heavy metals in their cytosol. This process is known as
Fig. 6.2 Mycoremediation of contaminants

6.6 Mycoremediation 115
Table 6.3 Mycoremediation of heavy metals and hydrocarbons polluted soils

Mushroom spp. Contaminant(s) Reference(s)
Pleurotus ostreatus Cd Favero et al. (1990a, b)
Hg Mandal et al. (1998)
Cd, Hg Yayçinkaya et al. (2002)
Pb Xiangliang et al. (2005)
Cd Das et al. (2007)
Cr Javaid and Bajwa (2007)
Cu, Cr, Ni, Zn Javaid and Bajwa (2008)
Co Xiangliang et al. (2009)
Pb Tay et al. (2009)
Crude oil Olusola and Anslem (2010)
Pb Liew et al. (2010)
Cd Tay et al. (2011)
Zn Jibran and Milsee Mol
(2011)
PAH Okparanma et al. (2011)
Cd Tay et al. (2011)
Cu, Cr, Ni, Zn Javaid et al. (2011)
Radioactive waste Eskander et al. (2012)
Cu, Cr, Fe, Zn Arbanah et al. (2012)
Heavy metals Oyetayo et al. (2012)
Cr Puentes-Cárdenas et al.
(2012)
Pb Prasad et al. (2013)
Cu, Cd, Fe, Ni, Pb, Lamrood and Ralegankar
Zn (2013)
Cu, Pb de Almeida and Burgess
(2013)
Cr Arbanah et al. (2013)
Oxo-biodegradable da Luz et al. (2013)
plastic
Pb Dulay et al. (2015)
Mn Wu et al. (2016)
Coriolus versicolor MKACC 52492 PAH Jang et al. (2009)
Schizophyllum commune, Polyporous sp. Malachite green Rajput et al. (2011)
Lentinula edodes 2,4-dichlorophenol Tsujiyama et al. (2013)
(DCP)
Agaricus bisporus, Lactarius piperatus Cd (II) ions Nagy et al. (2013)
Fomes fasciatus Cu (II) Sutherland and Venkobachar
(2013)
Pleurotus platypus, Agaricus bisporus, Cu, Zn, Fe, Cd, Pb, Lamrood and Ralegankar
Calocybe indica Ni (2013)
Flammulina velutipes Cu Luo et al. (2013)
Absidia cylindrospora Fluorene Garon et al. (2004)
Coniothyrium sp., Fusarium sp. PAHs Jacques et al. (2008)
mycofilteration, which is actually responsible for mycoremediation (Sesli et al.

2008; Falandysz et al. 2012). The success of mycoremediation depends on the capa-
bility of a fungal species to accumulate a particular toxic metal in its roots. A major-
ity of the mushroom species produce organic acids and extracellular enzymes
(peroxidases, manganese dependent peroxidase, lignin peroxidase, pectinases, oxi-
dases, xylanases and cellulases) that break-down the complex molecules into small
soluble molecules for mushroom nutrition and are considered as good recyclers in
nature (Mai et al. 2004; Kuforiji and Fasidi 2008; Zhu et al. 2013; Kulshreshtha
et al. 2014). This interesting feature of the mushrooms has gained attention in the
field of waste management as well as soil bioremediation (Akinyele et al. 2012;
Kumhomkul and Panich-pat 2013; Lamrood and Ralegankar 2013; Kulshreshtha
et al. 2013, 2014;). Mushrooms can accumulate large volume of noxious heavy met-
als [mercury (Hg), copper (Cu), chromium (Cr), cadmium (Cd), selenium (Se),
nickel (Ni), lead (Pb), zinc (Zn), manganese (Mn), iron (Fe) in their fruiting bodies
as compared to fruits, vegetables, and plants (Demirbas 2002; Cocchi et al. 2006;
Dogan et al. 2006; Zhu et al. 2011; Quarcoo and Adotey 2013). High metal concen-
tration has been reported in edible mushroom species such as Agaricus campestris,
Coprinus comatus, Calvatia utriformis, Lepista nuda (Kalac and Svoboda 2000;
Demirbas 2001; Falandysz et al. 2003; Ita et al. 2006; Garcıa et al. 2009). Various
fungal species like Agaricus bisporous, Phellinus badius, Phellinus sanguineus
have been reported to accumulate Cd (Zhu et al. 2011); Macrolepiota procera
(Scop.) Singer accumulated Pb (Arvay et al. 2014); Stropharia rugosoannulata,
Lactarius quietus, Psilocybe cubensis, Schizophyllum commune, Panellus stipticus
accumulated Hg (Arvay et al. 2014; Gabriel et al. 2016);
Sarcosphaeera crassa accumulate As (Konuk et al. 2007); Russula delica, Suillus
grevillei accumulated Cr (Cayır et al. 2010; Arvay et al. 2014); Boletus pulverulen-
tus, Cantharellus cibarius accumulated Cu (Arvay et al. 2014); Boletus pulverulen-
tus, Lactarius quietus, Macrolepiota procera accumulated Zn (Arvay et al. 2014)
and Agaricus arvensis, Agaricus porphyrizon, Amanita pantherina, Tricholoma vir-
gatum, Boletus edulis, Macrolepiota procera accumulated Se and Fe (Tuzen et al.
2007). Arbuscular mycorrhizal fungus (AM fungi) assisted phytoremediation is an
interesting approach to plant-based environmental remediation. In soil, AM fungi
are of great importance for heavy metal availability as they provide a direct connect-
ing link between roots and soil. Mycorrhiza can be defined as a symbiotic associa-
tion of fungus and plant root that is primarily responsible for nutrient transfer
(Brundrett 2004; Chibuike 2013). Mycorrhizal fungi expand the roots surface area
and thus increase the nutrients and water absorbance ability of roots, while the plant
supply carbohydrates to the fungi (Harrier and Watson 2004; Chibuike 2013).
Under heavy metal stress conditions, plant performance can be increased in
terms of lipid peroxidation, antioxidant enzyme and soluble amino acid on inoculat-
ing the plants with Mycorrhizal fungi (Andrade et al. 2009; Punamiya et al. 2010;
Achakzai et al. 2012). AM fungi protects the plants from toxic metals by accumulat-
ing toxic metals in their vacuole and binding them in their cell wall and also enhance
6.7 Conclusions 117
the plant growth in the polluted soils by improving mineral (phosphorous, nitrogen
and potassium) nutrition (Vivas et al. 2003; Bheemareddy and Lakshman 2011).
AM associations are important for the restoration of the degraded ecosystems
because of their benefits to symbiotic partners as they not only facilitate the plant
establishment and survival in contaminated soil but also provide protection to the
plants against phytotoxic effects of contaminants. The fungi enhance soil bioreme-
diation process by activating microbial activity and by improving soil structure
(Lenoir et al. 2016).
6.7 Conclusions
Contaminated soil is one among the major environmental issue due to its various
mutagenic and carcinogenic properties. Microbial remediation is a biological strat-
egy which has been applied for soil remediation and has various advantages over the
traditional remediation methods which include its low cost, high efficiency and
complete soil remediation.
Fungi are considered as the natural decomposers of ecosystem as they can effi-
ciently degrade and reduce the toxic heavy metals and other organic contaminants.
Rate of mycoremediation can be enhanced with the aid of certain carbon sources.
White rot fungi are the most important one as they can degrade different types of
toxic compounds such as heavy metals, aromatic hydrocarbons, chemical contami-
nants, dyes etc. Further studies are required so as to understand in depth mechanism
of pollutant degradation.
Deinococcus radiodurans is an important soil bacterium which has recently
gained the attention of scientists as it is capable to degrade highly radioactive tolu-
ene and mercury. This bacterium has the potential to degrade the radioactive waste
materials and to remediate the radioactive waste environments of energy, medicine
and military areas. Till date, genomic data of various microbes has been sequenced
and can be found on open databases. These databases must be analyzed thoroughly
so as to identify some novel contaminant degrading gene(s) and to develop geneti-
cally transformed strains of bacteria for efficient and complete remediation of con-
taminated soils. However, biosafety issues related to transformed microbes should
be taken into consideration. Another aspect which should be taken care is that the
transformed microbes can interact with the indigenous population of soil flora
which may affect the ecological diversity of the soil. Thus, the major focus of the
scientists should be to enhance the activity of transformed bacteria as well as to
maintain the indigenous microbial communities so that there is no loss to ecological
diversity. Furthermore, various chemical or physical methods could be implemented
in combination with microbial remediation so as to enhance the rate of soil
remediation.
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Chapter 7
Soil Remediation Through Algae, Plants
and Animals
Abstract There is an expanding enthusiasm for utilizing algal species for remedia-
tion of organic (hydrocarbons) and inorganic pollutants (heavy metals) in soils.
Several algal species have been isolated which have potential to remediate the pol-
luted soils and wastewaters. Phytoremediation mechanisms, although not quick but,
have the potential to restore the polluted soils. This strategy is centred on the mutual
act of plants and the microbial communities related with them to remove, degrade,
transform or immobilize the noxious contaminants present in sediments, soils, sur-
face water and ground water. Even though various chemical, physical, and biologi-
cal methods have been developed and suggested but, phytoremediation is the ideal,
innovative, as well as the safest method for removing toxic metals without any
noticeable side effects. Some lower animals take in heavy metals from soil and fur-
ther degrade, remove or lower their toxicity. It has also been reported that earth-
worm has significant potential to augment the stable fraction and to retard mobile
fraction of heavy metals. This chapter discusses the advantanges and the concerns
related to soil remediation mediated through aglae, plants and animals.
Keywords Heavy metals · Phytoremediation · Hyperaccumulator · Rhizofiltration

· Phytotransformation · Phytostabilization
7.1 Introduction
Recently, bioremediation has gained interest of the scientists for soil remediation
which involves the use of algae (phycoremediation), plants (phytoremediation) and
animals to detoxify the soil contaminants, sludges, rivers and lakes (Cunningham
and Berti 1993; Lim et al. 2010; Ellis et al. 2012). Algae are efficient accumulators
of toxic heavy metals and other xenobiotic compounds (Suresh and Ravishankar
2004). Hence, the bioremediation potential of algae proves useful for the extraction
of toxic metals and contaminants.
Soil remediation can be achieved with the aid of plants. Several reports on phy-
toremediation are now avaible (Lim et al. 2010; Ellis et al. 2012). It is a cost-
effective strategy which takes advantage of accumulation-activity of the plants. The

130 7 Soil Remediation Through Algae, Plants and Animals
plants having this ability are called bioaccumulators or hyper accumulators. Above
all, soil animals also help to achieve the soil remediation and play a direct or indirect
role in soil remediation process. Soil reclamation through soil animals can take
place in either of the two ways: firstly, the soil animals can act as indicator of pol-
luted soil and can address the risk and toxicity of polluted soils and Secondly, the
animals can directly take part in reclamation process by accumulating the toxic
metals to enhance the microbial and metabolic activity of polluted soils. Thus, soil
bioremediation strategies ensure environmental protection by using natural
resources for sustainable remediation which is economically and environmentally
beneficial for the society.
7.2 Phycoremediation
There is an expanding enthusiasm among environmentalists for utilizing algal spe-

cies for remediation of organic (hydrocarbons) and inorganic pollutants (heavy met-
als) in soils. Several algal species have been isolated which have the potential to treat
contaminated soils and wastewaters (Table 7.1). (Olguin 2003; Perales-Vela et al.
2006; Wenzel 2009; Kidd et al. 2009; Khan et al. 2009; Uroz et al. 2009; Rajkumar
et al. 2010; Olguin, 2012). Algae is the choice for remediation program because of
their small size, less complex body and high biomass, and also these are fast accu-
mulators of toxic compounds. Under heavy metal stress conditions, certain defence
mechanisms have been reported in algae which include compartmentalization, bind-
ing protein synthesis (i.e. PCs (poly(-y-glutamyl-cysteinyl)glycines) and MTs
(metallothionein) and phytochelatin complex formation), and exclusion and trans-
port of metal complexes to the vacuoles (Grill et al. 1989; Ben-Chekroun and
Baghour 2013). In Chlorella vulgaris, proline concentration was reported to rise
under chromium (Cr) and copper (Cu) stress conditions (Krämer et al. 1996; Sharma
and Dietz 2006; Ben-Chekroun and Baghour 2013) while, in Thalassiosira
pseudonana and T. weissflogii, synthesis of phytochelatins was observed under metal
stress conditions (Payne 2000; Ahner et al. 2002; Ben-Chekroun and Baghour 2013).
During phycoremediation, algae undergo two types of mechanisms, i.e. complex
formation and exclusion (inhibit the entry of metal ions). Formation of complex
tends to decrease the toxicity of inorganic contaminants i.e. heavy metals (Cobbett
and Goldsbrough 2002; Perales-Vela et al. 2006). Various green algal species includ-
ing Cladophora and Enteromorpha have been identified for their phytoremediation
potential (Al-Homaidan et al. 2011; Ben-Chekroun and Baghour 2013). Chlorophya
and Cyanophyta have also been reported as hyper-absorbents and hyper-accumula-
tors for arsenic (As) and boron (B). Some algal colonies were reported to live in
symbiotic association with roots of Arundo donax L. for phytoremediation of As
(Mirza et al. 2010a, b). The benefit of using algae in phytoremediation is the high
algal biomass production which leads to increase in absorption and accumulation of
toxic pollutants. However, the metal sorption capacity of algal strains should be high
enough for efficient phycoremediation (Ben-Chekroun and Baghour 2013).
7.2 Phycoremediation 131
Table 7.1 Reports on phycoremediation of heavy metals and other organic compounds
Sr.
No. Algal species Contaminant remediated References
1. Ankistrodesmus amalloides DDT, Dieldrin and Photodieldrin Neudorf and
Khan (1975)
2. Phytoplankton Chlorinated hydrocarbons Harding and
Phillips
(1978)
3. Agmenellum Bisphenol, 14c Naphthalene Cerniglia et al.
quadruplicatum (1979)
4. Scenedesmus bijugatus, Monocrotophos Megharaj
Chlorella vulgaris et al. (1987)
5. Selenastrum capricornutum BaP Warshawsky
et al. (1995)
6. Anabaena and Nostoc, DDT [1,1,1-trichloro-2,2- bis Megharaj
Chlorococcum (p-chlorophenyl) ethane] et al. (2000)
7. Cladophora crispate Zn(II) Ozer et al.
(2000)
8. Chlorella miniata Ni(II) Wong et al.
Chlorella vulgaris (2000)
9. Chlorella vulgaris Cd(II) Aksu (2001)
10. Spirogyra sp. Cr(IV) Gupta et al.
(2001)
11. Chlorella vulgaris Ni(II) Mehta and
Cu(II) Gaur (2001)
12. Chlamydomonas reinhardtii Cd(II) Adhiya et al.
(2002)
13. Ulothrix zonata Cu(II) Nuhoglu et al.
(2002)
14. Dunaliella sp. Cr(IV) Donmez and
Aksu (2002)
15. Chlorella vulgaris Cu(II) Mehta et al.
(2002)
16. Consortium of Pseudomonas Phenanthrene Muñoz et al.
migulae and Chlorella (2003)
sorokiniana
17. Nannochloropsis spp. Bisphenol A (BPA; 2,2-bis Ishihara and
Chlorella gracilis (4-hydroxyphenyl)propane) Nakajima
(2003)
18. Ulva lactuca Hg(II) Zeroual et al.
(2003)
19. Chlorella vulgaris Ni(II) Al-Rub et al.
(2004)
20. Chlorella sorokiniana Ni(II) Akhtar et al.
(2008)
21. Laminaria japonica Pb(II), Zn(II), Al(III), Cd(II), Cr(II), Cu(II) Lee et al.
(2004)
(continued)

Sr.
22. Cladophora crispate Cu(II) Ozer et al.
(2004)
23. Chlorococcum sp., R-endosulfan, endosulfan sulfate Sethunathan
Scenedesmus sp. et al. (2004)
24. Chlorella fusca Bisphenol A (BPA; 2,2-bis Hirooka et al.
(4-hydroxyphenyl)propane) (2005)
25. Chlamydomonas reinhardtii Cr(IV) Arıca et al.
(2005)
26. Fucus vesiculosus Ni(II), Cu(II) Rincon et al.
(2005)
27. Sphaeroplea sp. Ni(II) Srinivasa Rao
et al. (2005)
28. Chlamydomonas reinhardtii Hg(II), Pb(II), Cd(II) Tüzün et al.
(2005)
29. Cladophora fracta Pb(II), Cd(II) Lamaia et al.
(2005)
30. Chlorella vulgaris Ni(II) Aksu and
D€onmez
(2006)
31. Fucus vesiculosus Pb(II), Cd(II) Rincon et al.
(2005)
32. Sphaeroplea sp. Cu(II) Srinivasa Rao
et al. (2005)
33. Chlorella vulgaris Cd(II) Aksu and
D€onmez
(2006)
34. Cladophora fascicularis Cu(II) Deng et al.
(2006)
35. Chlorella miniata Cr(III) Han et al.
(2006)
36. Laminaria japonica Pb(II) Luo et al.
(2006)
37. Spirogyra sp. Cr(III) Bishnoi et al.
(2007)
38. Cladophora fascicularis Pb(II) Deng et al.
(2007)
39. Ulva lactuca Cr(IV) El-Sikaily
et al. (2007)
40. Sargassum sp. Cu(II) Karthikeyan
Ulva fasciata et al. (2007)
41. Ascophyllum nodosum Zn(II) Romera et al.
Asparagopsis armata (2007)
Chondrus crispus
Codium vermilara
Fucus spiralis
Spirogyra insignis
(continued)
7.2 Phycoremediation 133

Sr.
42. Scenedesmus quadricauda, Fluoranthene, Pyrene Lei et al.
Chlorella vulgaris, (2007)
Selenastrum capricornutum,
Scenedesmus platydiscus
43. Ascophyllum nodosum Ni(II), Cu(II), Pb(II), Cd(II) Romera et al.
Asparagopsis armata (2007)
Chondrus crispus
Codium vermilara
Fucus spiralis
44. Chondrus crispus Cu(II) Romera et al.
(2007)
45. Spirogyra neglecta Pb(II), Cu(II) Singh et al.
(2007)
46. Chlorella sorokiniana Cr(III) Akhtar et al.
(2008)
47. Five different species of Fenamiphos (ethyl 4-methylthio-m-tolyl Cáceres et al.
Cyanobacteria and green isopropyl phosphoramidate) (2008)
algae
48. Chlorococcum sp., Organophosphorus pesticide, fenamiphos Cáceres et al.
Pseudokirchneriella (ethyl 4-methylthio-m-tolyl isopropyl (2008)
subcapitata phosphoramidate) and its metabolites
fenamiphos sulfoxide phenol, fenamiphos
sulfone, fenamiphos sulfone phenol
fenamiphos sulfoxide, fenamiphos and
phenol
49. Chlorella vulgaris Cr(IV) Gokhale et al.
(2008)
50. Fucus vesiculosus Pb(II), Cu(II) Mata et al.
(2008)
51. Ulva lactuca Cd(II) Sari and
Tuzen (2008)
52. Protothecazopfii Polyurethane Ueno et al.
(2008)
53. Fucus vesiculosus Au(III) Mata et al.
(2009)
54. Rhizoclonium Cr(III) Onyancha
heiroglyphicum, Spirogyra et al. (2008)
condensate
55. Scenedesmus incrassatulus Cr(IV) Jacome-Pilco
et al. (2009)
56. Ulothrix cylindricum As(III) Tuzen et al.
(2009)
57. Pseudokirchneriella Carbamazepine Vernouillet
subcapitata et al. (2010)
58. Selenastrum capricornutum Naphthalene, pyrene, benzene, toluene, Gavrilescu
phenanthrene (2010)
(continued)

Sr.
59. Scenedesmus obliquus GH2 Crude-oil Tang et al.
(2010)
60. Cladophora hutchinsiae Se(IV) Tuzen and
Sari (2010)
61. Scenedesmus quadricauda Fungicides (dimethomorph and Olette et al.
pyrimethanil) and herbicide (isoproturon) (2010)
62. Chlorella vulgaris U(VI) Vogel et al.
(2010)
63. Cladophora sp. Pb(II), Cu(II) Lee and
Spirogyra sp. Chang (2011)
64. Scenedesmus Obliquus Zn(II) Monteiro et al.
(ACOI598) (2011)
65. Scenedesmus vacuolatus Fluoxetine and its metabolites Neuwoehner
norfluoxetine, propranolol, lidocaine, and and Escher
trimipramine (2011)
66. Chlamydomonas reinhardtii Herbicide (fluroxypyr) Zhang et al.
(2011)
67. Chlorella vulgaris Tetracycline de Godos
et al. (2012)
68. Monoraphidium braunii Bisphenol Gattullo et al.
(2012)
69. Chlamydomonas reinhardtii Herbicide (prometryne) Jin et al.
(2012)
70. Pediastrum tetras, Herbicide (mesotrione) Moro et al.
Ankistrodesmus fusiformis, (2012)
Amphora coffeaeformis
71. Spirogyra neglecta Cu(II) Singh et al.
(2012)
72. Spirogyra sp. Cr(IV) Yaqub et al.
(2012)
73. Stigeoclonium sp. diatoms, Ibuprofen, acetaminophen caffeine Matamoros
Chlorella sp., et al. (2015)
Monoraphidium
7.3 Phytoremediation
Phytoremediation process depends upon the mutual activity of plants and the asso-
ciated rhizospheric microbial communities to remove, degrade, transform or immo-
bilize the toxic contaminants present in sediments, soils, surface water and ground
water. This process, although not quick but, has the potential to restore the polluted
soils (Atma et al. 2017). Although several chemical, physical, and biological meth-
ods have been established but, phytoremediation is the ideal, innovative, as well as
the safest method for removing toxic metals without any noticeable side effects
(Atma et al. 2017). It has gained success in the treatment of various contaminants
7.3 Phytoremediation 135
such as pesticides, chlorinated solvents, explosives, petroleum, radionuclides,

hydrocarbons and heavy metals, in the polluted soils (Eapen and D’Souza 2005;
Eapen et al. 2007). Soils polluted with heavy metals are one of the major ecological
related worries of the present-day world. Several strategies have been deployed to
relieve the environment from these metals howeve, majority of them are costly and
do not provide efficient outcome. Interestingly, phytoremediation technique can
efficiently remediate the contaminated soils (Table 7.2). Aromatics-polluted soils
are also of environmental concern because of their carcinogenic and mutagenic
properties (Xu and Zhou 2017). Microbes can improve the efficiency of remediation
process through several mechanisms. Table 7.3 shows plant/microbe mediated
remediation of hydrocarbon contaminated soils. The efficiency or the phytoreme-
diation potential of plants can be increased by the reinforcement of certain kinds of
plant growth promoting (PGP) bacteria such as Azospirillum brasilense, Bacillus
pumilus, Pseudomonas putida, A. brasilense, Enterobacter cloacae etc. (de-Bashan
et al. 2012; Thijs et al. 2016). It has been observed that in salt stress conditions the
plant growth-promoting rhizobacteria (PGPR) not only prevents inhibition of pho-
tosynthesis and membrane damage but also shows positive effects on plant biomass
and health (Gerhardt et al. 2017). Moreover, plants with large biomass help to pre-
vent the spread of toxic compounds via air and water erosion (Cherian and Oliveira
2005). Candidate plants for phytoremediation must have the characteristics like
extensive root system, ability to tolerate large amount of contaminant, high biomass
production, and to survive under the adverse stress conditions. As mentioned before,
certain factor such as suitable bacterial and plant species must be taken into consed-
eration to accomplish the high remediation success (Ali et al. 2013; Balliana et al.
2017; Marchand 2017). Phytoremediation phenomena comprises ans is related to
various processes including rhizofiltration, phytodegradation, phytovolatilization,
phytostabilization, phytoextraction/phytoaccumulation, phytomining and phyto-
stimulation (Fig. 7.1).
Phytoremediation involves the following methods:
7.3.1 Rhizofiltration
Rhizofiltration uses the roots of the plants to remediate polluted soil(s) and water(s)
through absorption, concentration and precipitation of the contaminants (Dürešová
et al. 2014; Mirza et al. 2014). This technique is more suitable for remediation of
inorganic and organic pollutants from waste water with the aid of aquatic plants
(Jena and Dey 2017). Actually, rhizofiltration involves both phytostabilization and
phytoextraction (Dhanwal et al. 2017). An ideal plant for rhizofiltration must have a
large surface area and significant root biomass (Chen et al. 2016a, b). The roots of
many terrestrial plants grown hydroponically, including Brassica juncea, Helianthus
annuus and certain grasses, significantly remove toxic heavy metals like Cd2+, Cu2+,
Pb2+, Ni2+ and Zn2+ from aqueous solutions (Akpor and Muchie 2010). Rhizofiltration
is not limited to aquatic species only; certain terrestrial species such as Amaranthus
Table 7.2 Reports on phytoremediation of toxic-metal contaminated soils
136
Experiment
Element Contaminant (oxidation state) Plant(s) used type Remediation success Reference(s)
Chromium Cr(VI) Eichhornia crassipes (water Lab 5000–6000 mg/kg of Cr was Lytle et al. (1988)
(Cr) hyacinth) accumulated after supplying
10 mg/l solution for a period of
14 days.
Cr(III) Oryza sativa L. Lab 109.44 mg/kg of Cr(III) was Mishra et al.
accumulated by the roots when (1997)
supplied with 25.0 mg/l of
Cr(III).
Cr (VI) 140.34 mg/kg of Cr(IV) was
7
accumulated by roots when

supplied with 25.0 mg/l of
Cr(IV).
Cr(IV) Convolvuls arvensis L. Field C. arvensis accumulated more Gardea-Torresdey
than 30,000 mg of Cr/kg of dry et al. (2004)
root tissue from agar-based
medium containing 20 mg/l of
Cr(IV).
Soil contaminated with 14, Two poplar clones Eridano Lab Maximum amount of Cr Sebastiani et al.
800 mg/kg Cr (Populus deltoides × (587 mg/kg) has been reported to (2004)
maximowiczii-clone) and I-214 P. be accumulated in the roots of
xeuramericana Eridano.
Cr-containing soil [30 mg Cr3+ Helianthus annuus Lab 0.25 mg/g of Cr accumulated in Turgut et al.
(CrCl3.6H2O)] the plant stem due to the addition (2004)
of 0.3 g/kg EDTA into the soil.
Soil contaminated with Cr Salix fragilis (“Belgisch Rood”) Green house Maximum amount of Cr Vandecasteele
(65–779 mg/kg) and Salix viminalis (“Aage”) (50.5 mg/kg) accumulated in the et al. (2005)
roots of S. viminalis.
Soil Remediation Through Algae, Plants and Animals
Experiment
Cr(IV) and Cr(III) Salsola kali Lab A maximum of 3000 mg/kg Cr Gardea-Torresdey
accumulated in the roots when et al. (2005)
supplied with 20 mg/l of Cr (IV).
Cr-contaminated Eutric Cichorium intybus, Beta vulgaris, Field Z. mays accumulated the Ciura et al. (2005)
Cambisols, Agricultural Cucurbita pepo, Hordeum vulgare, maximum Cr (16.6 mg/m2/year).
academy research station near Phaseolus vulgaris, Brassica
Krakow, Poland oleracea var. capitata, Zea mays,
Pastinaca sativa, Medicago sativa

Dipotassium chromate Spinach (Spinacia oleracea L.), Lab B. campestris L. removed the Dheri et al. (2007)
(K2Cr2O7) Fenugreek (Trigonella maximum amount of Cr
foenumgraecum L.), and Raya (2.52–15.62 mg/g) from soil.
(Brassica campestris L.)
Salt of Chromium Phyllanthus amarus, Lab P. amarus absorbed the Eddy and Ekop
Stachytarpheta indica, Murraya maximum amount of Cr (2007)
koenigii (25.21 mg/kg) from Cr
contaminated soil.
Agricultural soil near Oryza sativa Field Maximum amount of Cr Liu et al. (2007)
Zhengzhou City, People’s (2.73–3.28 mg/kg) accumulated
Republic of China in the roots of the plant.
Potassium dichromate Bacopa monnieri L. Lab B. monnieri L. effectively Shukla et al.
(K2Cr2O7) removed Cr (319.5 mg/kg) from (2007)
solution.
Leather tannery sludge Melia azedarach Linn., Leucaena Lab Maximum 3450 mg/kg of Cr has Sakthivel and
(Sembattu, Trichy, Tamil leucocephla, Azadiracha indica been accumulated in the leaves of Vivekanandan
Nadu, India) A. indica. (2009)
(continued)
137
138
Experiment
0.02 mg/kg Cr in soil Triticum aestivum, Brassica Field 50.62 mg/kg of Cr accumulated Chandra et al.
campestris in roots of T. aestivum and (2009)
60.26 mg/kg accumulated in the
roots of B. campestris.
Potassium dichromate Brassica juncea L. (Indian Lab 640–4190 mg/kg of Cr Diwan et al.
(K2Cr2O7) mustard) accumulated in the stem of the (2010)
plant.
Cr(III) and CrO42- Azolla pinnata Field 415–1095 mg/kg of Cr Rai (2010)
accumulated in the plant tissue
7
when supplied with 0.5–3.0 mg/l

of Cr.
Cr(VI) Cynodon dactylon, Echinochla Lab A maximum of 94.59 mg/kg of Sampanpanish
colonum, Vetiveria nemoralis Cr was accumulated by C. et al. (2010)
dactylon when supplied with
5 mg/kg of Cr.
Potassium dichromate Cyperus kylinga, C. rotundus, Field Highest concentration of Cr Sundaramoorthy
(K2Cr2O7) Ludwigia parvifloria and Marselia (275.5 mg/kg) was recorded in et al. (2010)
quadrifolia the roots of C. rotundus.
Potassium dichromate Typha Angustifolia, Cyperus Field Highest concentration of Cr Chandra and Yadav
(K2Cr2O7) Esculentus, and Phragmites (75.82 mg/kg) was accumulated (2011)
Cummunis by T. angustifolia.
Chromium(III) nitrate Populus alba and Morus alba Lab Highest concentration of Cr Rafati et al. (2011)
nonahydrate (CrN3O9.9H2O) (15.39 mg/kg) was found in the
fallen leaves of M. alba.
Cr(III) 15 μgl−1 Typha latifolia L., Scirpus Lab S. Americans removed the Santos-Díaz and
americans Pers. maximum amount of Cr Barrón-Cruz
(40.7 mg/kg). (2011)
Experiment
Cr (0, 50, 100, 200, 400, 600, Arundo donax Lab 262.6 mg/kg of Cr was Kausar et al.
and 900 μg/l) accumulated in the stem of the (2012)
plant when supplied with
900 mg/lof Cr.
Potassium dichromate Lemna gibba and Ceratophyllum Lab 2816 mg/kg of Cr was Abdallah (2012)
(K2Cr2O7·5H2O) demersum accumulated by C. demersum
when supplied with 15 mg/l of
Cr.
Potassium dichromate Green gram (Vigna radiata), Pearl Lab 5.56 mg/kg of Cr was Dheeba and
(K2Cr2O7) millet (Sorghum bicolour), accumulated by the roots of Z. Sampathkumar
Sunflower (Helianthus annuus), mays when supplied with (2012)
Maize (Zea mays), Ground nut different concentration of Cr
(Arachis hypogaea) (10–50 mg/kg).
National metallurgical Azolla pinnata and Lemna minor Lab 0.026 mg/l of Cr was Vaseem and
laboratory effluent accumulated by the plant when Banerjee (2012)
(Jamshedpur, India) supplied with 0.07 mg/l of Cr.
0.2 mg/lCr6+ Nitella pseudoflabellat Lab 79 mg/kg of Cr was accumulated Gomes and Asaeda
by plant when supplied with (2013)
0.2 mg/l Cr6+.
Potassium dichromate Pithecellobium dulce, Pongamia Lab P. glabra accumulated the Unnikannan et al.
(K2Cr2O7) glabra, Cassia auriculata maximum amount (70 mg/kg) of (2013)
chromium in roots when treated
with 50 mg/kg of Cr.
Contaminated water from Alnus acuminata Lab 71 mg/kg of Chromium Escobar and
upper, medium and lower accumulated in the stem of the Dussán (2016)
basin of Bagota river, plant.
Colombia
(continued)
139
140
Experiment
Pulp and paper mill effluents Water hyacinth (Eichhornia Site E. crassipes accumulated the Kumar et al.
crassipes Solms.), Water caltrop maximum amount (0.5 mg/kg) of (2016)
(Trapa natans L.) Cr in the roots.
Plant grown on fly ash dumps Ziziphus mauritiana Field Maximum 38.5 mg/kg of Cr has Pandey and Mishra
of Panki thermal power station, been accumulated in the roots of (2016)
Kanpur, Uttar Pradesh, India Z. mauritiana.
Contaminated soil sites, 15 different plant species Field Among the 15 different plant Yuan et al. (2016)
Haining Country, northeast of species maximum amount of Cr
Zhejiang Province, China (372 mg/kg) was accumulated by
7
Chenopodium rubrum L.
collected from HN-4 site.
Different concentration of red Chrysopogon zizanioides (L.) Lab 14.07 mg/plant of Cr Gautam and
mud in soil amended with accumulated in the roots of plant Agrawal (2017)
sewage sludge when supplied with 10% of red
mud.
Municipal waste water Trapa natans L. Site 0.3 mg/kg of Cr accumulated in Kumar and Chopra
collected from activated sludge the roots of the plant after 60 (2017)
of Sarai, Jwalapur, Haridwar, days of treatment with municipal
India waste water.
Contaminated untreated mine Eichhornia crassipes Lab 3.5 mg/kg of Cr (VI) Saha et al. (2017)
waste water (Cr VI) from accumulated in the roots of plant
Sukinda chromite mines,
Orissa India
Experiment
Nickel (Ni) Ultramafic soil of North Berkheya coddii Lab B. coddii extracted 168 kg/ha of Robinson et al.
central Italy Ni from contaminated soil. (1997)
Ni-contaminated soil of Italy Alyssum bertolonii and Field Both these plants accumulated Anderson et al.
Streptanthus polygaloides 13,400 and 17,400 mg/kg dry (1999)
weight of Ni.
Ni(NO3)2·6H2O Pelargonium sp. Lab Pelargonium sp. accumulated a Dan et al. (2002)
significant amount of Ni in its
roots (1190 mg/kg DW) and

shoots (2100 mg/kg DW).
Ni-contaminated soil Brassica juncea and Brassica Lab B. juncea accumulated the Panwar et al.
carinata maximum amount (34.02 mg/kg) (2001)
of Ni.
NiSO4.6H2O Salix acmophylla Boiss. Lab 746.3 mg/kg of Ni was Ali et al. (2003)
accumulated in the roots of plant
when supplied with 10000 mg/kg
of Ni.
Ni and different concentration Helianthus annuus L., and Hybrid Field Leaves of Helianthus Liphadzi et al.
of EDTA poplar (Populus deltoids Marsh. × accumulated 19.2 mg/kg of Ni. (2003)
P. nigra L.)
Ni-containing soils of Albania 145 wild plant species collected Field Two species Alyssum murale and Cullaj et al. (2004)
(South-eastern Europe) from Ni contaminated sites Alysum markgrafii accumulated
the maximum content of Ni that
is 25,500 mg/kg and 23,700 mg/
kg, respectively.
Ni-containing soil [30 mg Ni2+ Helianthus annuus Lab 0.05 mg/g of Ni accumulated in Turgut et al.
(NiSO4.6H2O)] the plant leaves due to the (2004)
addition of 0.3 g/kg EDTA into
the soil.
Ni-contaminated ultramafic Berkheya coddii Lab Large amount of Ni accumulated Mesjasz-
soil from Agnes mine and the in the leaves (18000 μg/g) of B. Przybyłowicz et al.
Songimyelo Game Reserve, coddii. (2004)
141
NE South Africa
(continued)
142
Experiment
Different Ni concentrations Hordeum vulgaris, Spinacea Lab 473 mg/kg of Ni was Giordani et al.
oleracea, Brassica juncea, accumulated in the roots of beans (2005)
Phaseolus vulgaris, Sorgum when supplied with 300 mg/kg of
vulgare, Solanum lycopersicum, Ni.
Ricinus communis
Different Ni concentration Bornmuellera tymphaea and Lab L. emarginata removed a Chardot et al.
Leptoplax emarginata maximum amount of Ni (2005)
(4591 mg/kg) from soil.
Ni-contaminated Eutric Cichorium intybus, Beta vulgaris, Field C. pepo accumulated the Ciura et al. (2005)
7
Cambisols, Agricultural Cucurbita pepo, Hordeum vulgare, maximum amount of Ni

academy research station near Phaseolus vulgaris, Brassica (13.2 mg/m2/year).
Soil contaminated with Ni Salix fragilis (“Belgisch Rood”) Green house Maximum amount of Ni (9.5 mg/ Vandecasteele
(34–46 mg/kg) and Salix viminalis (“Aage”) kg) accumulated in the roots of S. et al. (2005)
fragilis.
Nickel sulphate (NiSO4)2 Scirpus littoralis Field S. littoralis accumulated Bhattacharya et al.
56.37 mg/kg of Ni. (2006)
Salt of Ni Phyllanthus amarus, Lab P. amarus showed the maximum Eddy and Ekop
Stachytarpheta indica, Murraya potential for the absorption of Ni (2007)
koenigii (769.00 mg/kg).
Experiment
1240 mg/kg of Ni in soil Seven species of Alyssum Field A. bracteatum accumulated the Ghaderian et al.
maximum amount of Ni that is (2007)
(2300 mg/kg Ni).
50, 100 mg/kg Ni in soil Cynodon dactylon, Festuca Lab Accumulation of Ni is high in Soleimani and
arundinacea shoots (25 mg/kg dry weight) of Hajabbasi (2009)
C. dactylon.
135 mg/kg of Ni in soil Rubus ulmifolius Lab Maximum amount of Ni found in Marques et al.
the roots (48–151 mg/kg) of the (2009)

plant.
Soil containing Ni (1.63 mg/ Triticum aestivum, Brassica Field 16.80 mg/kg of Ni accumulated Chandra et al.
kg) campestris in the roots of wheat and (2009)
7.28 mg/kg in roots of mustard.
Ni2+ Eichhornia crassipes Lab The plant ash showed a Hussain et al.
maximum of 1.978 mg/kg Ni2+ (2010)
from contaminated soil and
water.
Ni-contaminated water Helianthus annuus L. Lab 55.82 mg/kg of total Ni was Mukhtar et al.
accumulated by the plant when (2010)
supplied with 30 mg/l of Ni.
10 g of ground solid Ni Berkheya coddii Lab B. coddii accumulated 168 mg/kg Keeling et al.
of Ni. (2003)
Nickel(II) Sulfate Hexahydrate Typha angustifolia, Cyperus Field A total of 71.61 mg/kg of Ni was Chandra and Yadav
(NiSO4.6H2O) esculentus, and Phragmites absorbed by C. esculentus when (2011)
cummunis supplied with 16.0 mg/l of Ni.
Nickelous nitrate, hexahydrate Populus alba and Morus alba Lab 11.14 mg/kg of Ni was Rafati et al. (2011)
(NiN2O6.6H2O) accumulated in P. alba when
supplied with 120 mg/kg of Ni.
(continued)
143
144
Experiment
Ni-contaminated soil from Phragmites australis, Miscanthus Lab Among the five plant types J. Li et al. (2011)
Uranium Mill Tailings floridulus, Parthenocissus serotinus accumulated the
Repository, South China quinquefolia, Juncellus serotinus maximum amount (25.2 mg/kg of
Ni), in roots.
Ultramafic soil of Serbia Alyssum murale Lab Maximum concentration of Ni Tumi et al. (2012)
was found in leaves (13,160 mg/
kg) of the plant.
Ni-contaminated soil Lepidium sativum Lab 10.8095 mg/kg of Ni was Mojiri et al. (2013)
(1–20 mg/kg Ni) accumulated when supplied with
7
20 mg/kg of Ni
Hoagland’s solution Sorghum bicolor and Carthamus Lab 0.09 g/plant of Ni was absorbed Al Chami et al.
(containing different tinctorius by the shoots of S. bicolor when (2015)
concentration of Ni) supplied with 5 mg/l of Ni.
Plant grown on fly ash dumps Ziziphus mauritiana Field A maximum of 42.5 mg/kg of Ni Pandey and Mishra
of Panki thermal power station, was accumulated in the roots of (2016)
Kanpur, Uttar Pradesh, India plant.
Pulp and paper mill effluents Water hyacinth (Eichhornia Site A maximum of 1.0 mg/kg of Ni Kumar et al.
crassipes Solms.) and Water caltrop was accumulated in the leaves of (2016)
(Trapa natans L.) E. crassipes.
Lead (Pb) Different concentration of red Chrysopogon zizanioides (L.) Lab 0.93 mg/plant of Ni accumulated Gautam and
mud in soil amended with in the shoots of plant when Agrawal (2017)
sewage sludge supplied with 10% of red mud.
Lead acetate (Pb (CH3COO)2. Salix acmophylla Boiss. Lab 1038.5 mg/kg of Pb was Ali et al. (2003)
3H2O) accumulated by the roots of the
plants when supplied with
10000 mg/kg of Pb.
Experiment
Pb-contaminated soil Brassica juncea Lab 677 mg/kg of Pb was Lim et al. (2004)
accumulated by the shoots of the
plant, when supplied with
different Pb concentration and
5 mmol/kg of EDTA.
Pb-contaminated ultramafic Berkheya coddii Lab Maximum 3.3 mg/kg of Pb was Mesjasz-
soil from Agnes mine and the reported to be accumulated in Przybyłowicz et al.
Songimyelo Game Reserve, leaves of plant. (2004)

NE South Africa
Pb-contaminated Eutric Cichorium intybus, Beta vulgaris, Field Among the nine plant types C. Ciura et al. (2005)
Cambisols, Agricultural Cucurbita pepo, Hordeum vulgare, pepo accumulated the maximum
academy research station near Phaseolus vulgaris, Brassica amount Pb (12.6 mg/m2/year).
Lead nitrate (PbNO3)2 Scirpus littoralis Field S. littoralis accumulated a Bhattacharya et al.
maximum of 93.8 mg/kg of Pb. (2006)
75 mg/kg Pb in soil Alternanthera phyloxeroides, Lab Alternanthera phyloxeroides Cho-Ruk et al.
Sanvitalia procumbens, Portulaca accumulated maximum (2006)
grandiflora (22.49 mg/kg of Pb) in its tissues.
Salt of Pb Phyllanthus amarus, Lab S. indica accumulate 5.40 mg/kg Eddy and Ekop
Stachytarpheta indica, Murraya of Pb when supplied with 6 mg/ (2007)
koenigii kg of Pb.
500 mg/l Pb Zea mays L. Lab 900 mg/kg of Pb was Hovsepyan and
accumulated by the roots of plant Greipsson (2005)
when supplied with 500 mg/lof
Pb and 2.5 mmol/kg of EDTA.
(continued)
145
146
Experiment
Soil samples mixed with Pb Cynodon dactylon, Festuca Lab C. dactylon accumulated 15 mg/ Soleimani and
solution arundinacea kg of Pb in its roots when Hajabbasi (2009)
supplied with 100 mg/kg of Pb.
Contaminated soil in Portugal Rubus ulmifolius Lab 1178 mg/kg of Pb was absorbed Marques et al.
by the roots of R. ulmifolius (2009)
when supplied with 1400 mg/kg
of Pb.
0.43 mg/kg Pb in soil Triticum aestivum, Brassica Field 17.8 mg/kg accumulated in roots Chandra et al.
campestris of wheat and 26.14 mg/kg (2009)
7
accumulated in the roots of

mustard.
Lead nitrate (Pb (NO3)2) Tithonia diversifolia and Lab Helianthus annuus accumulated Adesodun et al.
Helianthus annuus maximum concentration of Pb in (2010)
roots that is 100.3, 96.8, 65.8 mg/
kg when supplied with 400 mg/
kg of lead nitrate in soil.
Pb-contaminated soil Zea mays Lab 14.17 mg/kg of Pb was absorbed Mojiri (2011)
by the roots of Z. mays when
supplied with 24 mg/kg of Pb.
Pb-contaminated water Helianthus annuus L. Lab 18.67 mg/kg of Pb was Mukhtar et al.
accumulated in the roots of H. (2010)
annuus when supplied with
30 mg/l of Pb.
Mining area in Hamedan 12 different plant species Field 10,121 mg/kg of Pb accumulated Nouri et al. (2009)
Province in the central west in the shoots of Echinophora
part of Iran platyloba.
Experiment
Pb(NO3)2 (1.0, 5.0, and Carex pendula Lab Maximum amount of Pb Yadav et al. (2011)
10.0 mg/L) in waste water accumulated in the roots
(1600 μg/g dry weight) of C.
pendula, when supplied with
10 mg/l of Pb(NO3)2.
Lead nitrate Pb (NO3)2 Typha angustifolia, Cyperus Field 92.8 mg/kg of Pb was Chandra and Yadav
esculentus, and Phragmites accumulated by P. cummunis (2011)
cummunis when supplied with 33.92 mg/l of

Pb (NO3)2.
Pb-contaminated soil from Phragmites australis, Miscanthus Lab 1.87 mg of Pb was absorbed by Li et al. (2011)
Uranium Mill Tailings floridulus, Parthenocissus P. australis when supplied with
Repository in South China quinquefolia 0.01–0.2 μg/l of Pb.
Pb(II) 60 μg/l Typha latifolia L., Scirpus Lab 148.3 mg/kg of Pb was absorbed Santos-Díaz and
americans Pers. by the roots of S. americanus Barrón-Cruz
when supplied with 60 μg/l of Pb. (2011)
Lead nitrate Pb (NO3)2 Lemna gibba and Ceratophyllum Lab 14.4 mg/l of Pb was accumulated Abdallah (2012)
demersum by L. gibba when supplied with
15 mg/l of Pb.
Toxic effluent from national Azolla pinnata and Lemna minor Lab 0.1 mg/l of Pb was accumulated Vaseem and
metallurgical laboratory, by L. minor when supplied with Banerjee (2012)
Jamshedpur 0.655 mg/l of Pb.
(continued)
147
148
Experiment
Natural wetland, located near 25 different plant species Site Among the 25 different plant Mazumdar and
river Barak in the Panchgram species maximum amount of Pb Das (2015)
area, Hailakandi district, Barak (197.66 mg/kg) was accumulated
Valley, Southern Assam, India in the leaves of Fimbristylis
bisumbellata.
Hoagland’s solution containing Sorghum bicolor and Carthamus Lab S. bicolor accumulated the Al Chami et al.
different concentrations of Pb tinctorius maximum amount of Pb (2015)
(90 mg/l) when supplied with
100 mg/l of Pb.
7
Effluents from Sagar Pulp and Water caltrop (Trapa natans L.) Site Maximum 0.65 mg/kg of Pb Kumar et al.
paper mills Ltd. Manglaur, and water hyacinth (Eichhornia accumulated in the roots of E. (2016)
Haridwar, India crassipes Solms.) crassipes.
Plant grown on fly ash dumps Ziziphus mauritiana Field A maximum of 45.33 mg/kg of Pandey and Mishra
of Panki thermal power station, Pb accumulated in the roots Z. (2016)
Kanpur, Uttar Pradesh, India mauritiana.
Lead nitrate (0,0.5,1.0 mg/kg) Brassica juncea Parents, B. juncea Green house Maximum amount of Pb (2.5 mg/ Rahman et al.
cv. Rai (P1), B. juncea cv. (hydroponics) kg) accumulated by the roots of (2016)
BARI-11(P2) and Their F1 Hybrids 2nd parent when supplied with
[(Rai♀ X BARI-11♂) and 1.0 mg/kg of Lead nitrate.
(BARI-11♀ X Rai♂)]
Experiment
Contaminated soil sites, 15 different plant species Field Among the 15 different plant Yuan et al. (2016)
Haining Country, northeast of species maximum amount of Pb
Zhejiang Province, China (1648 mg/kg) was accumulated
by Aster subulatus (Michx.) Hort.
ex Michx. collected from HN-4
site.
Contaminated water from Alnus acuminata Lab A acuminata accumulated Escobar and
upper, medium and lower 135 mg/kg of lead in the roots of Dussán (2016)
basin of Bagota river, the plant.
Colombia
Different concentration of red Chrysopogon zizanioides (L.) Lab 1.87 mg/plant of Pb accumulated Gautam and
mud in soil amended with in the shoots of plant when Agrawal (2017)
sewage sludge supplied with 10% of red mud.
10,000 mg/kg Lead nitrate Helianthus annuus and Hydrangea Lab H. paniculata accumulated the Forte and Mutiti
(Pb(NO3)2) paniculata maximum amount of Pb (2017)
(780 mg/kg) in the shoots.
Municipal waste water Trapa natans L. Site T. natans accumulated 0.3 mg/kg Kumar and Chopra
collected from activated sludge of Pb in the roots. (2017)
of Sarai, Jwalapur, Haridwar,
India
Manganese Mn-contaminated Eutric Cichorium intybus, Beta vulgaris, Field Medicago sativa accumulated Ciura et al. (2005)
(Mn) Cambisols, Agricultural Cucurbita pepo, Hordeum vulgare, 10218 mg/t f.w.; Hordeum
academy research station near Phaseolus vulgaris, Brassica vulgare accumulated 10120 mg/t
Krakow, Poland oleracea var. capitata, Zea mays, f.w.; Cucurbita pepo accumulated
Pastinaca sativa, Medicago sativa 66.5 mg/m2/year.
(continued)
149
150
Experiment
Soil contaminated with Mn Salix fragilis (“Belgisch Rood”) Green house S. fragilis accumulated the Vandecasteele
(292–803 mg/kg) and Salix viminalis (“Aage”) maximum amount of Mn et al. (2005)
(201.7 mg/kg) in the leaves.
Manganese chloride (MnCl2) Scirpus littoralis Field S. littoralis accumulated Bhattacharya et al.
494.92 mg/kg of Mn. (2006)
Salt of manganese Phyllanthus amarus, Lab P. amarus accumulated the Eddy and Ekop
Stachytarpheta indica, Murraya maximum amount of Mn (2007)
koenigii (111.28 mg/kg) when supplied
with 140 mg/kg of Mn.
7
Mn (50, 100 and 500 μM), Mn Juncus effusus L. Lab J. effusus accumulated Najeeb et al.
(500 μM) + citric acid (5 mM), 452.89 mg/kg of Mn in its roots (2009)
and Mn (500 μM) + EDTA when supplied with 500 μM Mn
(5 mM) and 5 μM EDTA.
2.0 mg/kg in soil Triticum aestivum, Brassica Field B. campestris the maximum Chandra et al.
campestris amount of Mn (287 mg/kg) in the (2009)
leaves but T. aestivum
accumulated the maximum
amount (84.1 mg/kg) in the roots.
MnCl2.4H2O Typha angustifolia, Cyperus Field 814.4 mg/kg of Mn was Chandra and Yadav
esculentus, and Phragmites accumulated by P. cummunis (2011)
cummunis when supplied with 20.54 mg/l of
Mn.
Mn (II) 1.8 mg/l Typha latifolia L., Scirpus Lab S. americanus accumulated the Santos-Díaz and
americans Pers. maximum amount (22.1 mg/kg) Barrón-Cruz
of Mn when supplied with (2011)
1.2 μg/l of Mn.
Experiment
Toxic effluents from National Azolla pinnata and Lemna minor Lab L. minor accumulated the Vaseem and
metallurgical laboratory, maximum amount (0.301 mg/l) Banerjee (2012)
Jamshedpur of Mn.
Effluents from Sagar Pulp and Water caltrop (Trapa natans L.) Site E. crassipes accumulated the Kumar et al.
paper mills Ltd. Manglaur, and water hyacinth (Eichhornia maximum amount (1.9 mg/kg) of (2016)
Haridwar, India crassipes Solms.) Mn in the leaves.
Plant grown on fly ash dumps Ziziphus mauritiana Field Z. mauritiana accumulated a Pandey and Mishra
of Panki thermal power station, maximum of 636 mg/kg of Mn in (2016)

Kanpur, Uttar Pradesh, India the roots.
Different concentration of red Chrysopogon zizanioides (L.) Lab C. zizanioides accumulated Gautam and
mud in soil amended with 20.93 mg/plant of Mn in the Agrawal (2017)
sewage sludge shoots when supplied with 5% of
red mud.
Municipal waste water Trapa natans L. Site T. natans accumulated a Kumar and Chopra
collected from activated sludge maximum of 1.4 mg/kg of Mn in (2017)
of Sarai, Jwalapur, Haridwar, the leaves.
India
Silver (Ag) Secondary treated municipal Lemna gibba L. Field Lemna gibba accumulated Sasmaz and Obek
waste water from Elazig, 2303% of silver. (2012)
Turkey
Different concentrations of Pistia stratiotes Lab P. stratiotes accumulated Hanks et al. (2015)
silver ions and nanoparticle 0.127 mg/kg of Ag by the roots
when supplied with 2 mg/l of
media.
Brown field sites contaminated Salix miyabeana, Festuca Lab Among the four plant species B. Desjardins et al.
with heavy metals, Montreal arundinacea, Medicago sativa, juncea accumulated the (2016)
(Quebec, Canada) Brassica juncea maximum amount of Ag
(4.60 mg/kg) from the
contaminated soil.
151
152
Table 7.3 Plant/microbe-mediated remediation of hydrocarbons contaminated soils

Experiment
Contaminant Plant(s)/ microbe(s) used type Remediation success Reference(s)
2,4,6-trinitrotoluene (TNT) Methanococcus sp. (strain B) Lab The bacteria transform 100 ppm of TNT within Boopathy and
40–60 days of treatment. Kulpa (1994)
Polychlorinated Biphenyl (PCBs) Rhodococcus erythropolis strain Lab Strain TA421 is capable of degrading PCB. Chung et al.
TA421 (1994)
Polychlorinated Biphenyl (PCBs) Rhizobium sp. Lab Rhizobial strains catabolized polychlorinated Damaj and
biphenyls. Ahmad (1996)
Glycerol trinitrate Beta vulgaris Lab Glycerol trinitrate degraded by plant. Goel et al.
7
(1997)
TNT Hairy root cultures of M. spicatum, Field M. aquaticum and M. spicatum accumulated Hughes et al.
M. aquaticum and Catharanthus TNT. (1997)
roseus
Dibenzothiophene (DBT) Rhizobium meliloti Lab Rhizobium strain utilized upto 30% of initial Frassinetti et al.
DBT concentration. (1998)
Atrazine Atrazine degrading bacteria Lab Among all bacteria Pseudomonas sp. strain Newcombe and
ADP, mineralized 17–35% of the atrazine. Crowley (1999)
RDX and HMX Myriophyllum aquaticum, Lab RDX was removed by both of the plants. Bhadra et al.
Catharanthus roseus (2001)
PAH and PCB Solanum nigrum, Triticum aestivum, Lab Up to 90% of PAHs and 20% of PCBs were Kucerova et al.
Glycine max etc removed by wheat cells. (2001)
Phenol and Chlorophenols Daucus carota (Hairy root cultures) Lab Transformed roots removed more than 90% of Araujo et al.
phenolic compounds. (2002)
Simazine M. aquaticum, Cannax×hybrida Lab 4 weeks old M. aquarium was reported more Knuteson et al.
tolerant to Simazine. (2002)
Trifluralin, Lindane Secale cereale Lab Ryegrass accumulated a fix amount of Li et al. (2002)
Trifluralin and Lindane
Experiment
4-Chlorophenol 2,6- Carex gracilis and Juncus effusus Lab Plants tolerated 30 mg/l of 4-chlorophenol after Wand et al.
Dimethylphenol Naphthalene few weeks. (2002)
HMX Populus spp. Lab 57% of HMX was accumulated by the plants. Yoon et al.
(octahydro-1,3,5,7-tetranitro- (2002)
1,3,5,7-tetrazocine)
2,4-Dichlorophenol Hairy root cultures of Brassica Lab Roots remediate approx. 97–98% of Agostini et al.
napus 2,4-dichlorophenol within one hour. (2003)
Benzotriazoles Helianthus annuus Hydroponics Phytotransformation of Benzotriazoles. Castro et al.

(2003)
Ethylene dibromide and Leuceana leucocephala Lab Plant metabolized EDB and TCE. Doty et al.
trichloroethylene (2003)
Perchlorate N. tabacum Hydroponics Plant accumulated 10ppb to 100ppm of Sundberg et al.
under green Perchlorate from hydroponic solution. (2002)
house
TNT, PPB M. aquaticum Field Soil concentration for TNT and PPB decreased Sung et al.
250 μg/kg and 500 μg/kg respectively. (2003)
TCE Populus spp. cell suspension Lab Populous suspension cells transformed TCE. Shang and
Gordon (2003)
Hexahydro-1,3,5-trinitro-1,3,5- Crude cell extract of Clostridium Lab RDX was transformed into several polar Zhang and
triazine (RDX) acetobutylicum (ATCC 824) intermediates by crude cell extract. Hughes (2003)
Methyl tert butyl ether Populus spp. Lab Contaminant was taken up by plant and Ma et al.
volatilize to environment. (2004)
Bitumen Medicago sativa (alfalfa), Pot The rhizosphere microflora of alfalfa has been Muratova et al.
Phragmites australis (reed) reported to show higher degradative potential as (2003)
compare to rhizosphere microflora of reed.
Phenanthrene Sinapis alba (White mustard) and Lab Concentration of phenanthrene decreased upto Hynes et al.
Sphingomonas yanoikuyae 90% in soil when inoculated with S. (2004)
(Bacteria) yanoikuyae.
(continued)
153
154
Experiment
2,4,6-Trichlorophenol Spirodella punctata Lab S. punctata is capable to degrade Jansen et al.
2,4,6-Trichlorophenol (2004)
Chlorpyrifos Enterobacter Strain B-14 Lab Enterobacter sp. degrades chlorpyrifos Singh et al.
(250 mg/liter) in less than 2 days. (2004)
Diesel-fuel Thespesia populnea (milo), Cordia Greenhouse Milo significantly reduced the concentration of Tang et al.
subcordata (kou), Myoporum and lab hexadecane and pyrene by 33% and 42% (2004)
sandwicense (sandalwood) respectively.
Hexahydro-1,3,5-trinitro-1,3,5- Acetobacterium paludosum Lab A. paludosum degraded RDX (3 mg/l) within 9 Sherburne et al.
triazine (RDX) days and convert it into soluble radio-labeled (2005)
7
metabolites
RDX, TNT, HMX Populus spp. (P. deltoids × nigra Lab Bacteria transformed all the contaminants Van Aken et al.
DN34) and Methylobacterium sp (2004)
Creosote Cladosporium, Fusarium, Lab Mixed population of fungi was reported more Atagana et al.
Penicillium, Aspergillus and effective for removal (94.1%) of Creosote (2006)
Pleurotus
Total petroleum hydrocarbons Cyperus laxus Lam. Lab Phytoremediation extent of plants was 90% Escalante-
(TPH) 5 g TPH/kg of dry perlite when inoculated with hydrocarbon degrading Espinosa et al.
microbes. (2005)
Petroleum Three grass species ( Cyperus Greenhouse Among all species, B. brizantha caused an Merkl et al.
aggregatus, Brachiaria brizantha, approximately 15% higher dissipation of (2005)
Eleusine indica) and Three legumes contaminants than other species.
species (Calopogonium mucunoides,
Stylosanthes capitata, Centrosema
brasilianum)
DDT Brassica juncea, Cichorium intybus Lab Hairy roots successfully degraded DDT. Suresh et al.
(Hairy root cultures) (2005)
Experiment
Polycyclic Aromatic Basidiomycetes Fungi, and Lab 93.7% of pyrene was degraded by coculture of Arun et al.
Hydrocarbons (PAHs) Pseudomonas sp. Coriolus versicolor and Fomitopsis palustris (2008)
TNT 8 plant spp. Lab G. max assimilated maximum concentration of Adamia et al.
TNT (2006)
Phenol and chlorophenols Ipomea batatus, D. carota, Solanum Lab 98.6% of phenol was removed by S. aviculare Araujo et al.
aviculare and 83% of chlorophenol was removed by D. (2002)
carota
Crude oil spills Pseudomonas strain PSI Lab Pseudomonas strain reported to degrade upto Mittal and
70.69% of alkaneas and 45.37% of aromatics. Singh (2009)
Diesel oil Canavalia ensiformis Lab 17% of the plants survived in the contaminated Balliana et al.
soil. (2017)
Petroleum hydrocarbons Vetiveria zizanoides Lab Vetiver was found to tolerate 5% of Brandt et al.
contaminant. (2006)
Diesel Lolium multiflorum L. (ryegrass) Pot Ryegrass roots effectively increased the Kaimi et al.
biodegradation of diesel-contaminated soil upto (2006)
55%.
Petrol hydrocarbons (PHCs) and Thirteen different plant species Greenhouse Among all species, Pisum sativum L (Pea), Liste and Prutz
polycyclic aromatic hydrocarbons pot Lepidium sativum L. (cress), and Viola tricolor (2006)
(PAHs) L. (pansy) extracted the maximum (60%)
amounts of PAHs from soil.
Crude oil Lolium arundinaceum Schreb. Field After three years of study, it has been reported White et al.
(fescue) Lolium multiflorum L. that phytoremediation management of (2006)
(ryegrass) Cynodon dactylon L. contaminated sites through plants and fertilizer
(Bermuda grass) addition led to reduction of contaminants.
(continued)
155
156
Experiment
Diesel Lolium multiflorum L. (ryegrass) Pot Ryegrass roots have been reported to be Kaimi et al.
effectively increased the biodegradation of (2006)
diesel-contaminated soil upto 55%.
Petroleum Brachiaria brizantha (Hochst. ex Lab Degradation of total aromatics oil and grease Merkl et al.
A. Rich.) Stapf was higher in planted than in unplanted soil. (2006)
Phenol B. juncea, Raphanus sativus, Lab B. juncea showed higher remediation potential Singh et al.
Azadirachta indica, Beta vulgaris than other plants. (2006)
(Hairy root cultures)
2,4-Dinitrotoluene A. thaliana Lab Phytotransformation of 2,4-dinitrotoluene. Yoon et al.
7
(2006)
Triclosan Trametes versicolor Lab Complete dechlorination of triclosan (5 g/l) has Bokare et al.
been reported within 20 min. (2010)
Crude oil Cyperus brevifolius (Rottb.) Hassk Net house C. brevifolius decrease the concentration of Basumatary
crude oil up to 61.2% in TC (contaminated soil et al. (2012)
without fertilizer) and 86.2% in TA
(contaminated soil with fertilizer).
Chlorpyrifos Streptomyces sp. strain AC7, S. sp. Lab 90% of the contaminant was degraded by both Briceno et al.
strain AC5 the strains within 24 h of incubation. (2012)
2,3,7,8-tetrachlorodibenzo-p Sphingomonas wittichii Lab Byproducts were totally degraded by microbial Bokare et al.
dioxin metabolism. (2012)
PAH Mesorhizobium sp., Alcaligenes sp. Lab 20.2% and 35.8% of PAH was removed after 56 Mao et al.
and Bacillus sp. days of incubation period. (2012)
p-nitrophenol (PNP) Bacillus pantothenticus and Bacillus Lab Both the spp. successfully degraded PNP. Sreenivasulu
aminovorans et al. (2012)
2,4,6-trinitrotoluene Achromobacter spanius STE 11 Lab Within 20 h of treatment, complete degradation Gumuscu and
of TNT (100 mg/l) was achieved by A. spanius Tekinay (2013)
STE 11.
Experiment
Phenol Halomonas sp. strain PH2-2 Lab The bacterial strain was able to degrade the Haddadi and
phenol up to 1100 mg/l of phenol Shavandi
(2013)
Lindane Sphingomonas sp. Lab The degradation efficiency of Sphingomonas Singh et al.
reached upto 99% when used in combination (2013)
with nanoparticles.
Atrazine Arthrobacter sp. strain DAT1 Lab Significant degradation of Atrazine was Wang et al.
achieved within 2–3 days when augmented (2013)

with Arthrobacter sp.
p-nitrophenol (PNP) Arthrobacter 4Hb Lab Within 24 hours of incubation period 17% Zohar et al.
removal of PNP was observed. (2013)
Naphthalene and crude oil Streptomyces spp. AB1, AH4, and Lab AM2, AB1 and AH4 remove 81.03%, 82.36% Ferradji et al.
AM2 and 85.23% of naphthalene after incubation (2014)
period of 12 days.
Phenanthrene, Fluorene and Kocuria sp. P10 Lab Maximum 83.2% of fluorene was degraded by Sun et al.
Fluoranthene Kocuria sp. P10 within 14 days after 14 days (2014)
incubation period.
Mixture of lindane and Cr(VI) Amycolatopsis tucumanensis DSM Lab Streptomyces sp. M7 and A. tucumanensis Polti et al.
45259, Streptomyces spp. A5, A11, successfully remove lindane and Cr(VI) (2014)
M7, MC1 respectively.
Crude Oil Thirty-three species of ornamental Greenhouse A maximum of 45–49% reduction of Ikeura et al.
flowering plants hydrocarbon in soil was done by Mimosa (2016)
pudica (Mimosa), Zinnia profusion (Zinnia),
Gazania rigens (Gazania), and Ipomoea
quamoclit (Cypress vine) after 180th day.
Aroclor 1248 (PCB) Burkholderia xenovorans with nZVI Lab 90% of the degradation was observed with Le et al. (2015)
(1000 mg/L) nZVI (1000 mg/L) with Burkholderia
xenovorans
(continued)
157
158
Experiment
Naphthalene, Anthracene, Pyrene Gordonia sp. Poz20, Amycolatopsis Lab 100% of naphthalene, 37.87% of anthracene, Ortega-
and fluoranthene sp. Poz14, Rhodococcus sp. Poz54 25.10% of pyrene, and 18.18% of fluoranthene Gonzalez et al.
were degraded within 45-days. (2015)
Polycyclic aromatic hydrocarbons Plants: Lolium perenne (rye grass), Greenhouse The maximum metal phytoextraction (36.1% Chen et al.
and Cd, Zn Seduce alfredii Bacteria: Cd and 12.7% Zn) and PAH removal (96.4%), (2016a, b)
Microbacterium sp. KL5 and were obtained by interplanting ryegrass with
Candida tropicalis C10 Seduce alfredii with regular re-inoculation with
Microbacterium sp. KL5 and Candida
tropicalis C10 in the co-contaminated soil.
7
Total petroleum hydrocarbons Sorghum halepense (L.) Pers. and Lab A maximum of 32.13% of TPH removed by S. Alavi et al.
Aeluropus littoralis (Guna) Parl halepense (L.). (2016)
Pyrene Thalassospira sp. TSL5-1 Lab Strain TSL5-1 successfully degraded Pyrene Zhou et al.
from contaminated soil. (2016)
Pyrene and Ni Scirpus triqueter Lab The addition of PGPB increased the efficiency Chen et al.
of phytoremediation of pyrene and Ni in soil. (2017)
Atrazine Plant: Phaseolus vulgaris L. (bean) Greenhouse A maximum of 76.63% atrazine was removed Madariaga-
Bacteria: Trichoderma sp., and from soil by the BT (bean + Trichoderma sp) Navarrete et al.
Rhizobium sp. treatment in 40 days. (2017)
Monochlorobenzoate Pseudomonas sp. 2-CBA, Lab Three strains successfully catabolized Xu et al. (2017)
Pseudomonas sp. 3-CBA, and Monochlorobenzoate with different pathways.
Hydrogenophaga sp. 4-CBA
Experiment
Cypermethrin Biochar with immobilized bacteria Lab The treatment of biochar (0.5%) with Liu et al.
(Bacillus Zhanjiangensis strain immobilized bacteria achieved the maximum (2017)
TJTB48, Bacillus Pseudofirmus degradation rate (82.18%) of cypermethrin
TJTB58 and Oceanobacillus kimchii within 40 days.
TJTB66)
Tetrabromobisphenol Geobacter and Mycobacterium with Lab Degradation rate of Tetrabromobisphenol Yang et al.
compost of spent mushroom increased with an addition of spent mushroom (2017)
compost.
Polycyclic aromatic hydrocarbons Burkholderia, Nitratireductor, Lab Addition of Hydroxypropyl-β-cyclodextrin Gao et al.
Nevskia and Sulfuritalea (HPCD) enhanced the degradation rate of (2017)
bacterial consortium.
Pyrene Coriolopsis byrsina strain APC5 Lab, Field 96.1% of pyrene was degraded by C. byrsina. Agrawal and
Shahi (2017)
Asphaltenes in crude oil Pseudomonas aeruginosa strains Lab Both strains degrade oil asphaltenes (59–72%) Gao et al.
(Gx and Fx) and pure asphalt (∼10%) in crude oil. (2017)
159
Fig. 7.1 Different mechanisms associated with phytoremediation phenomenon
paniculatus, Helianthus annuus and Brassica juncea with large roots have signifi-
cant potential to accumulate inorganic contaminants like Cu and Pb (Rahman et al.
2013). Dürešová et al. (2014), evaluated the capability of Arundo donax to grow in
Cd and Zn-contaminated water for rhizofiltration of heavy metals and it has been
found that A.donax succefully adsorb these contaminants. Elias et al. (2014),
explained the remediation potential of water hyacinth for extraction of toxic heavy
metals contaminants such as Zn, Cr, Fe, Cd, Cu and B from ceramic wastewater and
it was found that Cr was accumulated more than other metals. The concentration of
metals in roots was almost ten times higher than stem and leaves. Pérez-Palacios
et al. (2017), used transgenic tobacco hairy roots to hyper accumulate high Cu con-
centrations from wastewaters. This technique has also been used to extract organic
contaminants like tri chloro ethylene, atrazine, dioxins, tetrachloroethane, nitrotolu-
enesanilines, metolachlor and other petroleum hydrocarbons (Dhanwal et al. 2017).
The positive aspects of this method are: (1) low operational cost, (2) plants can be
grown directly on contaminated water, (3) aesthetically pleasing and after harvest-
ing, and (4) plants can be a substitute for biofuels. Furthermore, some drawbacks
are also present such as: (1) only small concentration of pollutants can be extracted
from media, (2) roots are not able to extract the pollutants which are present below
the root zone, and (3) proper disposal of harvested plants is again a major issue.
However, regular disposal and harvesting of plants are the main limitations of rhi-
zofiltration (Nikolic and Stevovic 2015).
7.3.2 Phytotransformation
Phytotransformation also termed as phytodegradation involves degradation of

organic pollutants by plant metabolic processes or by the effect of plant exudates.
Hence, the organic contaminants are broken down into smaller molecules which
integrate with plant tissues and enhance the plant growth. LeFevre et al. (2015)
observed the potential of Arabidopsis for phytotransformation of xenobiotic con-
taminants into less toxic compounds. Major limitation of this method is that pollut-
ants that enter the food chain require more than one growing season for efficient
transformation. Phytotransformation can be described under the following three
heads:
7.3.2.1 Degradation
Phytodegradation involves the degradation of metal pollutants which are accumu-

lated by plants via metabolic processes, or enzymatic breakdown of pollutants sur-
rounding the plants. The plant enzymes involved in the breakdown of the
contaminants are dehalogenases, phosphatases, dehydrogenases, reductases, oxy-
genases and peroxidases. In this way, the organic contaminants are degraded into
smaller molecules and then incorporated into plant parts for their growth. This
method is used to remove the volatile liquids, petroleum and aromatic compounds
from contaminated soils (Newman and Reynolds 2004). Some plant species
degrade the metal pollutants after absorbing them. Studies suggest that Raphanus
sativus can dregrade the phenols (Bhatti et al. 2012), Allium sativum (Osuji et al.
2014) and Solanum tuberosum (Kurnik et al. 2015) can degrade the metal contami-
nants using peroxidases. Plants also degrade the pollutants in association with sym-
biotic microbes (most commonly Pseudomonas and Bacillus spp. (Ahemad and
Kibret 2014). The phytodegradation potential is affected by the following: (1) poor
association of microorganisms and pollutants, (2) higher concentration of inor-
ganic salts, and (3) toxicity of chlorinated and long chain hydrocarbons to micro-
bial strains.
7.3.2.2 Volatilization
Plant leaves have the ability to transpire a large amount of water through stomata
that are present over the leaf surface. Phytovolatilization involves translocation of
metal contaminants by plants, followed by transpiration to the ecosystem (Vara
Prasad and de Oliveira Freitas 2003). In the atmosphere, these contaminants may
get degraded by the sunlight (photodegradation). It also involves the transformation
of heavy metals to volatile or gaseous state, with the help of certain root exudates
(Kumar et al. 2014). Some heavy metals pollutants such as selenium (Se), mercury
(Hg) and other volatile organic compounds (VOCs) are also reported to release
through the leaves into atmosphere (Sarkar et al. 2007). Water contaminated with
tritium can also be remediated by the phytovolatilization. Phytovolatilization also
plays an important role for remediation of ammonium (Wiessner et al. 2013).
Genetically modified plants, Nicotiana tabacum and Arabidopsis thaliana, have
also been reported to transform the inorganic and organic salts of Hg into volatile
form (Dhir 2013). The advantage of this technique is that it has been used for the
transformation of Hg into its less toxic form (elemental Hg) in the soil. However,
precipitation and deposition of mercury back into oceans and rivers from the envi-
ronment is the limitation of this phenomenon (Mahmoud and Hamza 2017).
7.3.2.3 Stabilization
This phenomenon involves immobilization of the heavy metal contaminants in the

soil. The immobilization can be achieved by decreasing the wind-blown dust or by
reducing soil erosion. The main purpose of stabilization is to increase the degrada-
tion of heavy metal pollutants, certain pesticides and other hydrocarbons (Callender
et al. 2016). Plant roots perform stabilization by inhibiting water percolation in soil
matrix, prevent direct contact with polluted soil and thus, retard soil destruction and
dispersal of the contaminants (Singh and Prasad 2015). Thus, phytostabilization is
economical, simple and eco-friendly strategy which stabilizes and reduces the bio-
availiability of pollutants (Jadia and Fulekar 2009). Generally, the plants with dense
root system, high transpiration rate, and canopies are preferred for phytostabiliza-
tion. Regular site monitoring should be performed to confirm optimum stabilizing
conditions. In case, when soil amendments are used to increase immobilization,
periodic reapplication of the amendments should be done to maintain the effective-
ness (Khan et al. 2012; Grobelak and Napora 2015). This technique is more suitable
to phytostabilize the heavy metal contaminants of waste landfill sites. Addition of
sewage sludge, straws and manure to the waste sites enhances the binding of toxic
metals to them. Limes and lime stones help to bind the cationic metals with the
inorganic wastes. Phytostablization is also efficient in stabilizing the sites contami-
nated with radionucleoides so as to hold the toxic pollutants in place and stop sec-
ondary contamination and its exposure to living beings. This technique is also
suitable for the extraction of As, Cd, Zn and U from contaminated areas (Sinha et al.
2009). An experiment was conducted so as to evaluate and compare the phytostabi-
lization capability of Spartina maritime and Spartina densiflora for Cr, Co and Ni.
It was observed that both the species significantly removed Co from the metal con-
taminated soil (Cambroll et al. 2011). The benefit of this treatment is that the dis-
posal of toxic compound is not essential and is reliable for quick immobilization of
soil pollutants (USEPA 2000). However, this method is only applicable for the areas
with low contamination.
7.3.3 Phytoextraction/Phytoaccumulation
Roots of the plants can uptake translocate and accumulate metal and radioactive
pollutants from soil and fresh or polluted water. The roots are capable to absorb both
organic as well as inorganic contaminants. A significant amount of metals translo-
cates via xylem tissues and accumulate in the leaves and shoots of plant (Singh et al.
2015). In a recent report Eichhornia crassipes was reported to significantly accumu-
late toxic Cr(VI) from the contaminated water thus, exhibiting its phytoremediation
potential (Saha et al. 2017). In a study on A. thaliana using caesium (Cs) and stron-
tium (Sr), majority of the Cs-137 accumulated in the roots, while 80% of the Sr-90
accumulated in the shoots of the plant (Kanter et al. 2010). Phytoaccumulation
depends upon two main factors i.e. the availability of metal in soil and the rate of
translocation from roots to upper parts of the plant. Furthermore, the availability of
the metals depends on water or soil conditions like lipophilicity, pH and clay con-
tent. The addition of certain soil amendments into the soil enhanced the metal avail-
ability in soil, more than 10 folds for cesium (Cs) and 100 folds for uranium (U) and
lead (Pb) (Huang et al. 1998). This technique has been successfuly used by
Phytotech. Inc. USA for extraction of cadmium (Cd) and lead (Pb). Moreover, this
technique also successfully removes selenium (Se) from Se contaminated soil
(Garousi et al. 2016). Lemna gibba was reported to accumulate significant amount
of zinc (Zn) from Zn contaminated water. High removal efficiency (25 mg g−1) and
accumulation capacity of L. gibba makes it a suitable plant for soil remediation
(Khellaf and Zerdaoui 2009). Petroselinum crispum accumulated mercuric chloride
in its leaves, stem and roots from Hg-contaminated soil. The rate of accumulation
was observed to be directly proportional to the concentration of the mercuric chlo-
ride in the soil (Bibi et al. 2016). The accumulation and uptake of mercuric chloride
was also evaluated in roots, stem and leaves of tomato plants and it has been reported
that higher concentrations of mercuric chloride (150 mg kg−1 soil), resulted in the
maximum accumulation in root, stem and leaves of tomato plants, while its lower
concentrations (10 mg kg−1 soil) did not show any significant accumulation. Two
aquatic plant species Eichhornia crassipes and Centella asiatica were evaluated for
their phytoaccumulation potential. C. asiatica removed 99.6% of Cu while, E.
crassipes removed 97.3% of Cu from copper solution. This indicates significant
phytoremediation potential of both the species (Mokhtar et al. 2011). Lactuca sativa
was also used as a model plant to analyze its phytoaccumulation potential and it was
observed that it efficiently accumulates Cd and Pb from contaminated water into its
edible parts. However, the accumulated Cd content was higher than other the other
metal (Rashid et al. 2014). An experiment was conducted in order to assess the
accumulation of As, Pb and Cd by Brassica juncea (Rai and BARI-11) parents and
their F1 hybrids. Out of the three metals, As was found in roots of the plants while
Pb was detected both in roots as well as the arial plant parts of F1 hybrids (Rahman
et al. 2016). It has been concluded that the cost of this technique is almost 10 times
low in contrast to other conventional methods of soil remediation (Salt et al. 1995).
However, in-depth study of the physiology, mechanism of contaminant uptake and
translocation and accumulation by the plants is required.
7.3.4 Phytomining
Plants absorb pollutants from the soil via their extensive root system and accumu-
late in their tissues. Bioharvesting of these accumulated pollutants from the plants
is termed as phytomining. Inadequacy and cost of conventional mining methods
have promoted the advantage of phytomining for metal extraction from their respec-
tive ores (Giurco and Cooper 2012; Wilson-Corral et al. 2012; Sheoran et al. 2013).
Ni is one of the metal which has gained maximum attention because of its high
commercial demand and availability of large Ni-rich land area (Bani et al. 2015;
Nkrumah et al. 2016). The presence of element(s) rich soils, hyper-accumulator
plants, optimized agronomic practices to maximize annual biomass-metal(s) yield,
efficient methods for recovery of element(s) from the post-harvest biomass and the
markets for the element(s) recovered from biomass, are the pre-requisites for suc-
cessful phytomining (Chaney et al. 2007; Bani et al. 2015). The results obtained
from the researches of previous two decades show that phytomining is a cost-
effective and feasible technique to remediate the contaminated soils. However, this
technique is still in its infancy and it is crucial to have a clear understanding of the
processes that are undergoing within the hyper-accumulators for acquisition, trans-
location and accumulation, for strategic remediation of polluted sites (Jan et al.
2014). Its progress, sustainability and commercial application can be achieved by
using it in combination with several other scientific advances (Novo et al. 2017).
7.3.5 Phytostimulation
It is the phenomenon in which the compounds released from roots of the plants
enhance the activity of rhizospheric microbes (Pilipović et al. 2015). Thus, it is basi-
cally a type of symbiotic relationship between microbes and plants where plants
provide nutrients to microbes and microbes provide a suitable environment for root
growth (Eskander and Saleh 2017). The principal mechanisms which are adopted
for metal resistance are metal reduction, metal oxidation, demethylation, methyla-
tion, metal ligand degradation, metal-organic complexion, enzymatic reduction,
extracellular and intracellular metal sequestration, exclusion by permeability bar-
rier, metal efflux pumps and formation of metal chelators like biosurfactants and
metallothioneins (Panda and Dhal 2016). Mostly, Cyanobacteria are used for phyto-
stimulation, as they release certain kind of phytohormones into the soil which are
7.4 Genetic Engineering Approaches for Phytoremediation 165
then absorbed by plants and mediate phytostimulation (Singh et al. 2009; Beharti
2014). This technique is reliable for the elimination of pesticides, PAHs and chlori-
nated solvents from soil and sediments. Higher concentration of pollutants may also
prove toxic to the plants. Phytostimulation can prove to be a low cost and practica-
ble approach to extract the metal contaminants from the soil.
7.4 Genetic Engineering Approaches for Phytoremediation
It is now possible to incorporate gene(s) irrespective of their origin into plants to get
the desired characteristics. Genetic engineering approaches are used to introduce
more efficient accumulator genes into mild accumulator plants possessing the
desired phenotype (tallness, high biomass etc.) to turn them to hyperaccumulators
(Chaney et al. 2000). Similarly, highly effective accumulator genes from taller
plants may be inserted into natural plants/mild accumulators to increase their bio-
mass, thus improving their phytoremediation properties (Davison, 2005; Malik
et al. 2014). Interestingly, induction of hairy roots in the hyperaccumulator plants
showed higher efficiency for rhizofiltration of radionuclides (Nedelkoska and
Doran, 2000). Moreover, the enhanced phytohormone synthesis in transgenic plants
can increase the biomass of hyperaccumulators and thus enhance the process phy-
toremediation (Hedden and Phillips, 2000). Table 7.4 summarizes selected studies
on transgenic plants expressing genes for metal tolerance and xenobiotic com-
pounds degradation.
Phytochelatins (PCs) are a family of heavy metal-inducible peptides crucial for
reclamation of toxic heavy metals, that have been identified in plants and some
microbes. They are synthesized from glutathione (GSH) in the presence of enzyme
PC synthase and heavy metal ions. Zhu et al. (1999a), raised transgenic mustard
expressing the E. coli gshII gene encoding glutathione synthetase (GS) in the cyto-
sol. The transgenic plants accumulated three folds higher Cd (per shoot) than the
non-transgenic controls. Transgenic tobacco plants harbouring mouse metallothio-
nein (MT) gene were found tolerant to Cd stress and accumulated twenty percent
lower Cd than the untransformed controls. The trait was stably inherited even in T2
progeny (Maiti et al. 1989). Several MT genes from mouse (MTI), human (MTIA
and MTII), Chinese hamster (MTII), yeast (CUP I) and pea (psMTA) have been
transferred to N. tabacum, Brassica species and A. thaliana (Eapen and D’Souza
2005) for improved Cd tolerance. Transgenic A. thaliana transformed with PsMTA
gene from Pisum sativum accumulated more Cu than the non-transformed control
plants (Evans et al. 1992). Transgenic B. juncea overexpressing enzymes for phyto-
chelatin synthesis extracted more Cr, Cd, Zn, Cu and Pb than wild plants (Zhu et al.
1999a, b). Transgenic A. thaliana harbouring glutathione S-transferase gene from
tobacco showed improved Al and Cu tolerance (Ezaki et al. 2000). In a similar
report the expression of citrate synthase gene resulted in plants with enhanced Al
tolerance (dela Fuente et al. 1997). Transgenic rice containing nicotinamine amino-
Table 7.4 List of transgenic plants for metal tolerance and degradation of xenobiotic compounds
(phytoremediation)
Target
Gene(s) Source plant(s) Effect on plants Reference(s)
MT2 gene Homo sapiens Nicotiana Cd tolerance Misra and
tabacum L. Gedamu
and Brassica (1989)
napus L.
MTA gene Pisum sativum Arabidopsis Cu accumulation Evans et al.
thaliana (1992)
MT-1 gene Mus muridae N. tabacum Cd tolerance Pan et al.
(1994)
CYP105A1 Streptomyces N. tabacum Sulfonylurea resistance OKeefe et al.
griseolus 1994
CUP-1 gene Yeast Brassica Cd accumulation Hasegawa
oleracea var. et al. (1997)
botrytis
Ferretin Glycine max N. tabacum Increased accumulation Goto et al.
of Fe (1998)
FRE-1 and FRE-2 Yeast N. tabacum Increased accumulation Samuelsen
of Fe et al. (1998)
Nt CBP4 N. tabacum N. tabacum Increased Pb Arazi et al.
accumulation and Ni (1999)
tolerance
Pentaerythritol E. cloacae N. tabacum Degrade GTN (Glyceryl French et al.
tetranitrate trinitrate) and TNT (1999)
reductase (Trinitrotoluene)
Ferretin G.max Oryza sativa More accumulation of Goto et al.
Fe (1999)
CYP71A10 G. max N. tabacum Phenylurea tolerance Siminszky
et al. (1999)
Zn transporters A. thaliana A. thaliana Increased Zn Van der Zaal
ZAT(AtMTPI) accumulation et al. (1999)
Glutathione O. sativa B. juncea Increased Cd tolerance Zhu et al.
synthetase (1999a)
γ-Glutamylcysteine E. coli B. juncea Increased Cd tolerance Zhu et al.
synthetase (1999b)
CYP450 2E1 H. sapiens N. tabacum Oxidation of ethylene Doty et al.
dibromide and TCE (2000)
(Trichloroethylene)
Glutathione-s- N. tabacum A. thaliana Tolerance to Na, Al and Ezaki et al.
Transferase Cu (2000)
ACC-deaminase Bacteria Lycopersicon Increased accumulation Grichko et al.
esculentum of Ni, Cu, Zn, Mg, Cd, (2000)
Pb and Co
CAX-2 (Vacuolar A. thaliana N. tabacum Increased accumulation Hirschi et al.
transporters) of Mn, Cd and Ca (2000)
(continued)

Target
Glutathione B. juncea Increased Cd Pilon smits
reductase accumulation et al. (2000)
Nitroreductase E. cloaceae N. tabacum Degrade TNT Hannink et al.
(2001)
Cysteine synthase O. sativa N. tabacum Tolerance to Cd Harada et al.
(2001)
Pentaerythritol Enterobacter N. tabacum Metabolises Rosser et al.
tetranitrate cloacae pentaerythritol (2001)
reductase tetranitrate and
nitroglycerine
Nicotinamine amino Hordeum O. sativa Plant survive in Fe Takahashi
transferase (NAAT) vulgare L. deficient soil et al. (2001)
γ-glutamylcysteine E. coli A. thaliana Increased As Dhankher
synthetase ECS + accumulation and et al. (2002)
arsenate reductase tolerance
arsC
Arsenate reductase Bacteria B. juncea Increased tolerance to
γ-glutamylcysteine As
synthetase
CYP76B1 Helianthus N. tabacum, Increased herbicide Didierjean
tuberosus A. thaliana tolerance et al. (2002)
CYP2B6 H. sapiens O. sativa Metabolises benfuresate Kawahigashi
and ethofumesate et al. (2002)
γ-glutamylcysteine E. coli Populus Increased accumulation Koprivova
synthetase ECS tremula × P. of Cd in roots et al. (2002)
alba
Mn-peroxidase C. versicolor N. tabacum Degrade Limura et al.
pentachlorophenol (2002)
ACC deaminase E. cloacae B. napus Increased accumulation Nie et al.
and tolerance to As(V) (2002)
γ-glutamylcysteine E. coli A. thaliana Increased tolerance and Dhankher
synthetase ECS + and N. accumulation of Cd et al. (2003)
arsenate reductase tabacum
arsC
Phytochelatin T. aestivum N. glauca Increased accumulation Gisbert et al.
synthase (Ta PCS) of Pb (2003)
Pentaerythritol E. cloacae N. tabacum Degrade GTN (Glyceryl Hannink et al.
tetranitrate trinitrate) and TNT (2003)
reductase (Trinitrotoluene)
Znt A-heavy metal E. coli A. thaliana Tolerance to Pb and Cd Lee et al.
transporters (2003)
selenocysteine lyase M. musculus A. thaliana Increased tolerance and Pilon et al.
SL accumulation of Se (2003)
YCF1 Yeast A. thaliana Tolerance to Pb and Cd Song et al.
(2003)
(continued)

Target
CUP-1 gene Yeast N. tabacum Increased accumulation Thomas et al.
of Cu (2003)
Selenocysteine A. bisculatus A. thaliana Selenite Resistance Ellis et al.
methyl transferase (2004)
Acetyltransferase Noccaea A. thaliana Increased tolerance to Freeman et al.
SAT goesingense Ni (2004)
γ-Glutamylcysteine B. juncea B. juncea Increased tolerance to Flocco et al.
synthetase (ECS), metolachlor, atrazine, (2004)
Glutathione 1-chloro-2,4-
synthetase (GS) dinitrobenzene and C
phenanthrene
Cysteine synthase Spinacia N. tabacum Increased tolerance to Kawashima
oleracea Ni, Se, Cd and et al. (2004)
accumulation of Cd
selenocysteine Astragalus B. juncea Increased tolerance, LeDuc et al.
methyltransferase bisulcatus accumulation and (2004)
SMT volatilization of Se
ATP sulfurylase B. juncea Increase tolerance to Se Van Huysen
CAPS et al. (2004)
ATP sulfurylase A. thaliana B. juncea Increased Wangeline
APS1 concentrations of Cr, et al. (2004)
Mo, Cd, V, W and Cu in
shoots
Glutathione E. coli B. juncea Increased accumulation Bañuelos
synthase GS of Se in leaves et al. (2005)
ATP sulfurylase A. thaliana B. juncea Increased Se
APS accumulation in leaves
NA synthase NAS1 A. thaliana N. tabacum Increased Ni Douchkov
accumulation and et al. (2005)
tolerance
CYP2C9 H. sapiens O. sativa Tolerance to Hirose et al.
sulfonylurea (2005)
CYP1A1, CYP2B6, H. sapiens O. sativa, Resistance to herbicides
Inui and
CYP2C9 and Solanum including sulfonylurea Ohkawa
CYP2C19 tuberosum (2005)
NA synthase NAS1 Hordeum A. thaliana Increased tolerance to Kim et al.
vulgare Zn, Cd, Mn, Ni, Cu and (2005)
Fe
Glutathione G. max N. tabacum Increased tolerance to Karavangeli
S-transferase I alachlor et al. (2005)
Nitroductase E. coli A. thaliana Degradation of TNT Kurumata
et al. (2005)
CYP2B6 H. sapiens O. sativa Degradation of Kawahigashi
metolachlor et al. (2005a)
(continued)

Target
CYP1A1 H. sapiens O. sativa Degradation of Kawahigashi
simazine and atrazine et al. (2005b)
CYP2B22 Sus scrofa O. sativa Herbicides tolerance Kawahigashi
CYP2C49 et al. (2005c)
Peroxidase L. esculentum L. esculentum Degradation of phenol Oller et al.
(2005)
NA synthase NAS1 Noccaea A. thaliana Increased Ni Pianelli et al.
caerulescens accumulation and (2005)
tolerance
Laccase C. versicolor N. tabacum Degradation of Sonoki et al.
pentachlorophenol and (2005)
bisphenol A
ACC deaminase P. putida B. napus Increased accumulation Stearns et al.
and tolerance to Ni (2005)
Extradiol Terrabacter sp. A. thaliana Remediation of Uchida et al.
dioxygenase (Dbf 2,3-dihydroxybiphenyl (2005)
B)
Haloalkane Rhodococcus N. tabacum Dechlorination of 1
dehalogenase sp. chlorobutane
(DhaA)
Atrazine Pseudomonas N. tabacum, Degrade atrazine to Wang et al.
chlorohydrolase sp. Medicago hydroxyatrazine (2005)
(AtzA) sativa
CYP1A1, CYP2B6 H. sapiens O. sativa Degrade metolachlor Kawahigashi
and CYP2C19 and atrazine et al. (2006)
Selenocysteine Astragalus B. juncea Increased Se LeDuc et al.
methyltransferase bisulcatus and accumulation and (2006)
SMT and ATP A. thaliana tolerance
sulfurylase APS1
Phytochelatin A. thaliana N. tabacum Increased Cd Pomponi et al.
synthase PCS1 accumulation and (2006)
tolerance
xpl A Rhodococcus A. thaliana Degrades RDX (Royal Rylott et al.
rhodochorous Demolition Explosive) (2006)
selenocysteine lyase Astragalus B. juncea Increased accumulation Bañuelos
SL and bisulcatus and of Se et al. (2007)
selenocysteine Mus musculus
methyltransferase
SMT
rhlA and rhlB P. aeruginosa N. tabacum Increased tolerance to Brichkova
Al et al. (2007)
(continued)

Target
CYP450E1 H. sapiens N. tabacum Increased degradation Dixit et al.
of chloropyriphos and (2008)
anthracene
743B4, 73C1 A. thaliana A. thaliana Increased degradation Gandia-
of TNT Herrero et al.
(2008)
Protox B. subtilis O. sativa Increased tolerance to Jung et al.
diphenyl ether and (2008)
oxyflufen
CYP1A1 H. sapiens O. sativa Increased metabolism Kawagashi
of norflurazon and et al. (2007)
chlorotoluron
Biphenyl Burkholderia N. tabaccum Oxidized Mohammadi
dioxygenase gene xenovorans 4-chlorobiphenyls et al. (2007)
γ-glutamylcysteine E. coli B. juncea Increased accumulation Reisinger
synthase ECS and and tolerance to Pb, Cr, et al. (2008)
glutathione synthase Zn, As and Cd
GS
pnrA P. putida P. tremula ×P. Increased tolerance of Van Dillewijn
tremuloides TNT et al. (2008)
Acyl-CoA- binding A. thaliana A. thaliana Increased accumulation Xiao et al.
protein ACBP1 and tolerance to Pb (2008)
iaaM + ACC A. tumefaciens Petunia Increased accumulation Zhang et al.
deaminase and P. putida hybrida, N. of Cu and Co (2008)
tabacum
Metallothionein P. sativum P. alba Increased tolerance to Balestrazzi
MTA1 Zn and Cu et al. (2009)
merA E. coli Liquidambar Increased volatilization Dai et al.
styraciflua × and tolerance to Hg (2009)
L. formosana
Hg binding protein, Bacillus A. thaliana Increased accumulation Hsieh et al.
merP megaterium and tolerance of Cd, Pb (2009)
and Hg
Metallothionein A. thaliana A. thaliana Increased accumulation Rodríguez-
MT4 and tolerance of Zn and Llorente et al.
Cu (2010)
Glutathione Trichoderma N. tabacum Increased remediation Dixit et al.
transferase virens and tolerance of (2011)
anthracene
Metallothionein Cajanus cajan A. thaliana Increased Cd and Cu Sekhar et al.
MT1 accumulation and (2011)
tolerance
(continued)

Target
Human P450 2E1 Trichoderma Alfalfa plants Remediate TCE Zhang and
(CYP2E1) gene and virens, H. (pKHCG) (trichloroethylene) and Liu (2011)
glutathione sapiens Cd
S-transferase (GST)
Vacuolar H+ T. aestivum N. tabacum Increased Cd Khoudi et al.
pyrophoshatase VP1 accumulation and (2012)
tolerance
Metallothionein Pisum sativum P. alba Increased accumulation Turchi et al.
MTA1 of Zn and Cu (2012)
Metallothionein S. cerevisiae N. tabacum Increased Zn and Cd Daghan et al.
MTII accumulation (2013)
ScMTII S. cerevisiae N. tabacum Increased accumulation
of Cd
As(III) vacular Pteris vittata A. thaliana Increased tolerance and Chen et al.
antiporter ACR3 translocation of As (2013)
G protein DEP1 O. sativa A. thaliana Increased Cd Kunihiro et al.
accumulation and (2013)
tolerance
Metallothioneins B. campestris A. thaliana Increased Cd and Cu Lv et al.
MT1 and MT2 accumulation and (2013)
tolerance
Vacuolar Cd A. thaliana and N. tabacum Increased tolerance and LeBlanc et al.
transporter YCF1 S. cerevisiae accumulation of As (2013)
+Phosphate
transporter PHT1
and PHT7
ScYCF1 Yeast Populus alba Increased accumulation Shim et al.
× P. tremula of Zn, Pb, and Cd in (2013)
var. roots
glandulosa
Human P450 2E1 Trichoderma Alfalfa plants Degrade Hg and TCE Zhang et al.
(CYP2E1) gene and virens, H. (pKHCG ) (2013)
glutathione sapiens
S-transferase (GST)
Metallothionein Iris lactea var. A. thaliana Increased tolerance to Gu et al.
MT2b chinensis Cd (2014)
gshI E. coli Populus Increased GSH Gyulai et al.
canescens (Glutathione) (2014)
concentration
P450 CYP1A2 H. sapiens A. thaliana Resistance to Kebeish et al.
phenylurea and (2014)
chlortoluron
(continued)

Target
nidA, nidB, NahAa Pseudomonas A. thaliana Increased tolerance and Peng et al.
and NahAb putida G7, assimilation of PAHs (2014a)
Mycobacterium
vanbaalenii
PYR-1
NahAa, NahAb, P. putida G7 A. thaliana Improved uptake of Peng et al.
NahAc, and NahAd and O. sativa phenanthrene and lower (2014b)
concentrations in shoots
and roots
AtHMA4 A. thaliana N. tabacum Decreased Siemianowski
accumulation of Cd in et al. (2014)
shoots and roots.
Increased accumulation
of H2O2
bphC Bacteria N. tabacum Increased tolerance to Viktorova
PCBs et al. (2014)
Apoplastic A. thaliana A. thaliana Increased Cd Chen et al.
endo-β-mannase accumulation and (2015)
MAN3 tolerance
AtACBP1 and A. thaliana B. juncea Increased Pb Du et al.
AtACBP4 accumulation in roots (2015)
Metallothionein I. lactea var. A. thaliana Increased Cu Gu et al.
MT2b chinensis accumulation and (2015)
tolerance
IlMT2b I.lactea var. A. thaliana Increased concentration
chinensis of Cu and decrease in
H2O2
gshI E. coli Populus Increased accumulation He et al.
tremula × P. of Cd in shoots (2015)
alba
FDH Plant and A. thaliana Increased CN tolerance Kebeish et al.
(gi145358030), bacteria and metabolism (2015)
CYND (gi1167514)
Class 1 hemoglobin N. tabacum N. tabacum Decreased uptake of Cd Lee and
Hb1 and enhanced metal Hwang (2015)
tolerance
γ glutamyl-cisteine S. thermophilus Beta vulgaris Increased accumulation Liu et al.
synthetase- and tolerance to Zn, Cu (2015)
glutathione and Cd
synthetase GCS-GS
operon
CsGSTU1 and Citrus sinensis N. tabacum Tolerance to Lo Cicero
CsGSTU2 Fluorodifen et al. (2015)
dBphC Soil Medicago Increased tolerance to Wang et al.
metagenomic sativa L. cv. PCBs/2,4-DCP and (2015)
library Gongnong degradation of PCBs
(continued)

Target
OsMTP1 Oryza sativa L. N. tabacum Increased accumulation Das et al.
cv. IR64 of Cd (2016)
Transcription factor A. thaliana A. thaliana Increased accumulation Chen et al.
ZAT6 of phytochelatins and 2016a, b
tolerance to Cd
Arsenic Westerdykella A. thaliana Increased tolerance to Verma et al.
methyltransferase aurantiaca AsIII and AsV (2016)
(WaarsM) gene
transferase genes (NAAT) from barley overproduced iron chelator-deoxymugineic

acid in rice and grew well in Fe deficient soils (Takahashi et al. 2001). Overexpression
of bacterial 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase resulted in
enhanced tolerance to Cd, Co, Cu, Ni, Pb and Zn (Grichko et al. 2000). There are
reports on genetic manipulation of metal transporters, leading to transgenic plants
with modified metal tolerance/accumulation chacteristics. Transgenic A. thaliana
plants harbouring Zn transporter-ZAT gene from Thlaspi goesingense accumulated
two-fold Zn in the roots than the controls (Van der Zaal et al. 1999). Improved Ni
tolerance was observed in transgenic plants by the expression of transporter gene-
NtCBP4 (Arazi et al. 1999). Transgenic tobacco expressing A. thaliana calcium
vacuolar transporter CAX-2 gene were tolerant to increased Mn concentration
(Hirschi et al. 2000). In a similar report, overexpression of transgenic ABC trans-
porter yeast protein (YCF1) involved in the transfer of Cd into vacuoles in A. thali-
ana led to enhanced Pb and Cd tolerance (Song et al. 2003). Increased Fe tolerance
was observed by the overexpression of metal transporter AtNramp1 (Curie et al.
2000). There are several successful reports on raising transgenic plants for increased
iron uptake (Robinson et al. 1999); enhanced tolerance to Se (DeSouza et al. 2000)
enhanced tolerance to As (Dhankher et al. 2002) and phytoreduction of Hg (Bizily
et al. 2003). A. thaliana plants expressing MerA and MerB genes could grow on
fifty-fold higher methylmercury concentrations and volatilized the elemental mer-
cury, than the non-transformed control plants. Transgenic A. thaliana plants express-
ing two genes: E. coli arsenate reductase gene (ArsC) driven by soybean rubisco
promoter (SRS1p) and E. coli γ-glutamylcysteine synthetase gene (γ-ECS) driven
by actin promoter (ACT2p) gained 4 to 17-fold greater fresh shoot weight and accu-
mulated 2 to 3-fold more As per gram of tissue than the control plants expressing
γ-ECS or ArsC alone (Dhankher et al. 2002).
Each metal has a unique mechanism for uptake, translocation and sequestration
therefore, it is crucial to design appropriate strategies for producing transgenic
plants specific for a particular metal. Like others remediation strategies, phytoreme-
diation has its disadvantages which includes potential phytotoxicity of the contami-
nant, climatic and geological limitations, potentially longer timescale as compared
to other technologies and risk of the pollutants or its metabolites of food chain
contamination (Macek et al. 2000). The operational period of this technique is very
long (many years). Proper disposal and consumption of heavy metal containing
plants is also a major concern. Moreover, unfavourable climatic conditions can also
affect the plant growth, biomass production and remeditaion efficiency of the plants
(Jena and Dey 2017). Further research is still required to overcome these drawbacks
for effective implementation of this technique, in future.
7.5 Animal-Mediated Soil Remediation
Some lower animals adsorb heavy metals from soil and further degrade, remove or
lower their toxicity (Prakash et al. 2017) (Table 7.5). In a study by Wang et al.
(2007), it was concluded that a combination of earthworm and straw mulch can suc-
cessfully increase the Cu concentration in plants. The earthworm can effectively
accumulate Pb and its concentration can increase with an increase in Pb concentra-
tion of the soil (Kou et al. 2008).
During vermicomposting of animal manure, transformation of heavy metals to
less toxic byproducts has been reported Lv et al. (2016). It has also been reported
that earthworm has significant potential to enhance the stable fraction of heavy met-
als and to decrease the mobile fraction. Moreover, other animal classes like enchy-
traeids, collembola, mites, isopods, nematodes and protozoans have been deployed
for the assessment of polluted soil toxicity (Haimi 2000). The accumulation and
transfer of soil contaminants from animals, especially the earthworms to food web
is the biggest limitation of this strategy.
Table 7.5 Animal groups Species Animal group

associated with
Folsomia candida Collembola
bioremediation
Folsomia fimetaria
Orchesella cincta
Eisenia fetida/Andrei Earthworms
Lumbricus terrestris
Aporrectodea caliginosa
Enchytraeus albidus Enchytraeids
Porcellio scaber Isopods
Trichoniscus pusillus
Platynothrus peltifer Mites
Plectus parietinus Nematodes
Colpoda cucullus Protozoans
References 175
7.6 Conclusions
Severe contamination of soil with various organic and inorganic contaminants is

one of the major concerns of the society as these contaminants may enter into the
food chain or food web and can create serious health related issues. Organic con-
taminants and heavy metals are not easily degradable and difficult to dispose hence,
they can cause various mutagenic, teratogenic and carcinogenic effects in living
beings. Various techniques are availableand are being practised for soil remediation
but, among them phytoremediation has proved as an environment friendly and a
better substitute to other techniques. Choice of most suitable plant for remediation
as well as the introduction of some foreign genes can successfully enhance the
remediation potential of plants.
Furthermore, use of algal species is also an emerging tool for remediation which
is called Phycoremediation. With the aid of algal spp. various organic contaminants
can be degraded, besides that there are some pollutants which are not degraded by
them. Introduction of genetic engineering and transgenic approaches can enhance
the remediation potential of algal spp. No doubt, there is a bright future of soil bio-
remediation, besides that there is still a long way to go so as to comparatively study
all the remediation process as well as to evaluate their remediation potential.
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Chapter 8
Nanobioremediation
Abstract Soil and water contamination through heavy metals, hydrocarbons and
radioactive wastes is of global concern as these factors have cumulative effect on the
environment and human health. Removal of contaminants from polluted soils is one
of the major challenging tasks in the twenty-first century. Nanobioremediation is an
emerging technology for remediation of pollutants with the aid of biosynthetic
nanoparticles. Because of their unique chemical and physical properties, nanopar-
ticles have gained the attention of scientists from different fields of environmental
sciences. Nanobioremediation, an offshoot of nanotechnology, is a promising and
rapidly growing soil remediation technology. The present chapter summarizes the
synthesis of nanoparticles from yeast, fungi, bacteria and plants and their potential
uses for remediation of polluted soils and sludges. Nanobioremidiation can be
deployed where the other conventional remediation treatments do not prove to be
useful because nanoparticles are less toxic to soil flora and enhance the microbial
activity. Although various researches have been conducted on the physical and
chemical properties of nanoparticles still, more information is required about their
interaction, and adsorption with the contaminated soils. Further researches in soil-
bioremediation should focus on the combined use of nanoparticles, genetically
modified microbes and plants to design environment friendly, cost-effective, robust
and sustainable remediation strategies.
Keywords Nanotechnology · Bioremediation · Nanoparticles · Contaminants ·

Green synthesis
8.1 Introduction
The amount of wastes generated in the environment has been increasing day by day
as a result of the rapid advancement in science and technology (Dastjerdi and
Montazer 2010). Nowadays, the sustainability of environment is one of the major
concerns. Among various restoration strategies which have been discussed earlier,
bioremediation most commonly involves the use of microorganisms and plants.
However, these remedies are not feasible under highly toxic conditions. Nanoparticles

198 8 Nanobioremediation
have the promising potential to restore the toxic environments rapidly without
harming the surrounding micro flora.
The particles having size <100 nm are termed as nano or ultrafine particles.
These particles can be manufactured from various materials depending upon their
shape, size and chemical composition (Das and Ansari 2009). Nanoparticles can be
divided into two main group i.e. organic nanoparticles and inorganic nanoparticles.
Organic particles consist of carbon particles and inorganic particles consist of noble
metal particles, magnetic particles and various other semiconductor nanoparticles.
Incidental nanoparticles (coal and diesel combustion), natural nanoparticles (lunar
dust, volcanic eruptions) and engineered nanoparticles (metal-based materials) have
also been identified (Monica and Cremonini 2009). Nanoparticles can be synthe-
sized from different materials and through various conventional methods (Table 8.1).
These days, due to the cost effectiveness and efficiency of nanoparticles, they have
been successfully deployed for remediation of contaminated soil and water sources
(Friedrich et al. 1998; Dimitrov 2006; Dastjerdi and Montazer 2010).
Table 8.1 Synthesis methods for creating nanoparticles (Das and Ansari 2009)
Nanomaterials Methods Examples
Metal Photochemical Pt, Rh, Pd, Ir, Ag, Au, Cu, Co, Ni, Fe & Ni, Cu3Au,
nanoparticles electrochemical Co & Ni, Cd & Te, Cd & Se, ZnS
biochemical
thermochemical
Polymer NMs Electrochemical Nanowire of polypyrrole, polyaniline, poly(3,4-
polymerization ethylenedioxythiophane) dendrimers (PAMAM)
Carbon NMs Arc-discharge Cylindrical nanotube (SWNT, MWNT) fullerenes
Laser ablation
Chemical vapor
deposition
Nanocomposite Innovative methods Nanocomposite of polyethylene oxide and
polyethyleneimine; CNT epoxy composites include
hydrocarbon polymer composites, conjugated
polymer composites, CNTs with polycarbonates,
fluoropolymers, polyethylene glycol, polyester
polyamides, and so forth
Metal oxide Hydrothermal ZnO, Fe2O3, Fe3O4, MgO, BaCO3, BaSO4, TiO2
nanoparticles Solvothermal
Sol–gel
Reverse micelles
method
Electrochemical
deposition
Bionanomaterials Biological Viruses, plasmids, and protein nanoparticles
8.2 Properties of Nanoparticles 199
8.2 Properties of Nanoparticles
Physical properties of nanoparticles depend upon various parameters which include

size, shape, state of size distribution, structure, surface area and solubility. Among
these shape and size are the two most important factors which defines the nanopar-
ticles. Nanoparticles of different shapes and sizes can be produced such as spherical,
tube form, rod and disk shaped and other forms like urchins, squares, ellipsoids and
worms are also reported. Increase in the surface to volume ratio of nanoparticles
makes their surfaces more available for reactions as the number of molecules pres-
ent at the particle’s surface increases exponentially. Nanoparticles of size less than
20–30 nm are characterized by an excess of energy at the surface and are thermody-
namically unstable because of the interfacial tension that leads to a reduction in the
surface area. The optical properties of nanoparticles also vary according to their size
and shape. The surface area and surface morphology of nanoparticles can be exam-
ined with the use of BET (Brunauer–Emmett–Teller) and SEM (scanning electron
microscope). NTA (nanoparticle tracking analysis) method is widely used for analy-
sis and visualization of nanoparticles present in liquid medium. This technique
allows us to find the size distribution profile of nanoparticles with diameter ranging
from 10–1000 nm in a liquid medium (Filipe et al. 2010). Chemical properties of
nanomaterials include surface chemistry, chemical composition photocatalytic
properties and zeta potential (Rao and Biswas 2009). Zeta potential reflects the elec-
tric potential of nano-particles. The nano-particles having zeta potential more than
(+/−) 30 mV are stable in suspension because the surface charge inhibits the aggre-
gation of nanoparticles in this form. Quantum dots consist of different types of
metal complexes like magnetic transition metals or semiconductors with diameter of
size less than 10 nm. The activity of quantum dots can be enhanced by the coating
of their surfaces with some biocompatible material or some suitable functional
groups. Nanoparticles also have distinct mechanical properties which can be deter-
mined through various parameters such as hardness, adhesion, elastic modulus, fric-
tion, stress and strain. Atomic force microscopy (AFM) is a tool which is used to
measure the friction and adhesion between solid surface and nanoparticles.
Furthermore, coagulation, surface coating and lubrication also aid to mechanical
properties of nanoparticles (Guo et al. 2014). Metal nanoparticles also possess
higher thermal conductivities as compared to other solids or fluids. For example
thermal conductivity of copper nanoparticles is 700 times and 3000 times higher
than that of water and engine oil respectively. Nanofluids are prepared by mixing of
nanoparticles with liquids such as ethylene glycol, oils or water. These fluids exhibit
excellent thermal properties as compared to other fluids due to their large surface
area. It has also been found that nanofluids prepared from CuO or Al2O3 in ethylene
or water exhibit very high thermal conductivity (Cao 2002). Nanoparticles also pos-
sess optical properties as they are small enough to confine their electrons and pro-
duce quantum effects (Hewakuruppu et al. 2013). For example, gold nanoparticles
appear deep-red to black in solution because of quantum effect. Gold nanoparticles
of size 2.5 nm melt at much lower temperatures (~300 °C) than the gold slabs
Fig. 8.1 Types of nanoparticles (a) Iron oxide nanoparticles, (b) Zero valent iron nanoparticles,
(c) Fe/Pd nanoparticles (d) Nano-clay polymer, (e) Carbon nanotubes, (f) Silica-PAMAM
Dendrimer nanoparticles, (g) Iron-oxide magnetic nanoparticles, (h) Nanosponge for pollutants
removal
(1064 °C). Nanoparticles absorb the solar radiations to a great extent as compared
to other materials due to their large surface area. An application of metal nanopar-
ticles in the area of optics is termed as ‘Plasmonics’. There are various advantages
of nanomaterial over the micro particles as they have very large surface area/unit
mass. Large amount of contaminated material comes into their contact which
enhances its activity. Due to the quantum effect exhibited by nanoparticles, less
activation energy is required to make reactions feasible. Another property which is
exhibited by nanoparticles is surface plasmon resonance which can be used for the
detection of heavy metals. Various metallic and nonmetallic sources can be used to
manufacture different types of nanomaterials of different shapes and sizes (Fig. 8.1).
Nanoparticles can easily penetrate into polluted area where other microparticles
cannot reach. Furthermore, nanoparticles can be applied for both in situ and ex situ
remediation treatments. They can be easily used in slurry reactor as well as can be
used with a solid matrix (Friedrich et al. 1998; Dimitrov 2006; Dastjerdi and
Montazer 2010).
8.3 Types of Nanoparticles
8.3.1 TiO2 (Titanium Dioxide) Based Nanoparticles
TiO2 is commonly used material in electronics, semiconductors, energy convertors,

and photocatalytic and gas sensors. Because of its availability and low toxicity, TiO2
particles are used as a photocatalyst for the treatment of polluted water. ‘Reverse
micelle’ technique has been successfully used to produce TiO2 nanoparticles so as
8.3 Types of Nanoparticles 201
to adsorb various dyes from liquid media (Stathatos et al. 1999). Nbutoxymethyl-
2-chloro-2, 6-diethylacetanilide, an organic contaminant found in agricultural soil
can be efficiently removed from soil with the use of immobilized TiO2 nanoparticles
(Mahmoodi et al. 2007). Pollutants which have been successfully removed by TiO2
based nanoparticles are enlisted in Table 8.2.
8.3.2 Iron Based Nanoparticles
Nanoscale iron particles represent latest and highly efficient environmental reme-
diation technologies which can prove as cost-effective solutions to some most chal-
lenging environmental cleanup problems. Nanoscale iron particles have large
surface areas and high surface reactivity. Stabilized ZVI nanoparticles have been
examined for their potential to immobilize or remove various pollutants (organic
and inorganic) from contaminated soil and water. Various reports are available on
use of zero-valent iron (ZVI or Fe0) for extraction of pollutants from solid or liquid
media. In case of chlorinated contaminants, ZVI removed them by reductive dechlo-
rination reaction. Iron nanoparticle having size 10–30 nm have been used for the
extraction of Pb and Cr from aqueous solution (Ponder et al. 2000). In a study, As
(V) and As (III) have been strongly absorbed by nZVI from contaminated soil, with
the addition of Iron (hydr) oxides in nZVI (Kanel et al. 2005). Zhang et al. (2010)
compared the effectiveness of FeS, starch-stabilized ZVI and magnetite nanoparti-
cles for immobilization or removal of As from two Ascontaminated soils. It has
been found that all the nanoparticles effectively reduce the concentration of As in
polluted soils. It has also been reported that CMC-stabilized Fe-Pd nanoparticles
double the degradation rate of TCE from polluted soil as compare to starch-stabilized
nZVI. The enhanced degradation rate of CMC-stabilized ZVI is due to the high
specific reactivity and ability to stabilize the nanoparticles (He et al. 2007).
8.3.3 Bimetallic Nanoparticles
Although ZVI have the potential to degrade various polychlorinated bipehenyls,

chlorinated aromatics and aliphatic compounds besides that there are certain limita-
tions of ZVI technique (Wang and Zhang 1997) such as production of chlorinated
byproducts and decrease the reactivity of iron with respect to time. To overcome
these challenges iron can be used in combination with other metals such as Zn, Ni,
Cu, Pd with a higher catalytic activity than iron. Palladium is the most common
reductive dehalogenation catalyst used with nZVI for remediation purposes. PBDEs
(Polybrominated diphenyl ethers) are now ubiquitous in the environment, with soil
as a major sink, especially in e-waste recycling sites. In a study by Xie et al. 2014,
Ni/Fe bimetallic nanoparticles were used to degrade decabromodiphenyl ether pres-
ent in soil. It has been found that these bimetallic nanoparticles successfully degrade
Table 8.2 Nanoparticle mediated removal of contaminants

Type of nanoparticle Contaminant Reference
Arginine-modified TiO2 Nitrobenzene Makarova et al.
(2000)
Anatase TiO2 Phenol Andersson et al.
(2002)
TiO2 nanoparticle Rhodamine 6G Chen et al. (2003)
TiO2/SrFe12O19 composite Procion Red MX-5B Fu et al. (2006)
Mesoporous Anantase nanocrystal Methyl orange Wang et al. (2004)
Mesoporous TiO2 nanopowder3 Rhodamine B Peng et al. (2005)
Mesoporous titania nanohybrid 4-Chlorophenol, methyl Paek et al. (2006)
orange
Rutile TiO2 nanoparticle Parathion Wang et al. (2006)
TiO2/AC nanoparticle Methyl orange Li et al. (2006)
TiO2 nanoparticle Basic dye Wu et al. (2006)
TiO2 nanoparticle and carbonized Benzene, toluene Chuang et al. (2007)
composite
Zero-valent powder iron Nitrate Choe et al. (2000)
Colloidal zero-valent powder iron Herbicide Molinate Feitz et al. (2005)
Iron sulfide nanoparticle Lindane Paknikar et al. (2005)
Zero-valent powder iron Arsenic(V) Kanel et al. (2006)
ZVI nanoparticle Perchlorate Xiong et al. (2007)
ZVI Humic acid Giasuddin et al.
(2007)
Iron nanoparticle Ni(II) Li and Zhang (2007)
Pd/Fe nanoparticle Trichloroethene Wang and Zhang
(1997)
Lien and Zhang
(2001)
Ni/Fe nanoparticle Trichloroethene Schrick et al. (2002)
Ni/Fe nanoparticle Carbon tetrachloride Feng and Lim (2005)
Cu/Fe nanoparticle Nitrate Liou et al. (2005)
Fe-nanocomposite Azo-dye orange(II) Feng et al. (2003)
Dithiocarbamateanchored nanocomposite Pb(II), Cd(II), Cr(III) Say et al. (2006)
CNTs Pb(II) Li et al. (2002)
Li et al. (2005)
CNTs KMnO4 oxidized Cd(II) Li et al. (2003a, b, c)
Single-walled carbon nanotubes Zn(II) Lu and Chiu (2006)
Multi-walled carbon nanotubes
Al2O3/CNTs Fluoride Li et al. (2001)
Aligned carbon nanotubes Fluoride Li et al. (2003a, b, c)
CNTs (graphitized) 1,2-Dichlorobenzene Peng et al. (2003)
CNTs Trihalomethanes (CHCl3) Lu et al. (2005)
Poly(amidoamine)dendrimer Cu(II) Diallo et al. (1999)
Octylurea DAB-32 Pyrene Arkas et al. (2003)
(continued)
8.3 Types of Nanoparticles 203

Type of nanoparticle Contaminant Reference
Mag-Im18 Cu(II) Takafuji et al. (2004)
Modified jacobsite (MnFe2O4) Cr(VI) Hu et al. (2005a, b)
Maghemite nanoparticle
Magnetic chitosan nanoparticle Co(II) Chang et al. (2006)
Magnetic alginate microcapsules Ni(II) Ngomsik et al.
(2006)
PAA (poly amidoamine) bound iron oxide Methylene blue Mak and Chen
(2004)
Bromelain Chen and Huang
(2004)
the pollutants with 72% removal efficiency at pH 5.6 and a dosage of Ni/Fe 0.03 g/g.
Another report by Schrick et al. 2002 it was reported that degradation potential of
Fe/Pd nanoparticles was nine times more as compared to nZVI for degradation of
TCE.
8.3.4 Nanoclays
Clays are the materials with several layers which can absorb +ve and –ve ions and
undergo various ionic interactions with absorbed ions. Due to their extensive sur-
face area, nano sized dimensions, nanoclays can take up the contaminants from
polluted soil or water. However, lack of permanent porosity is a major disadvantage
of nanoclays. Ding et al. 1999 successfully removed TiO2 from liquid media by
dispersing them in layered nanoclays. Various reports are available for the use of
nanoclays for removal of various contaminants. The removal potential of allophane
nanoclay was investigated by Yuan and Wu 2007, and it was reported that allophane
nanoclay successfully reduced the concentration of phosphorus from 14.2 to
4.2 mg/L (70%).
8.3.5 Nanotubes
Nanotubes are the small hollow cylindrical materials which consist of carbon nano-
tubes and fullerenes. Nanotubes have various unique properties including high
strength, electrical and thermal conductivity, high adsorption capability and stiff-
ness (Yang 2003). Nanotubes have pores over their surface on which the molecules
can adsorb and interact with nanotubes. This interaction depends upon the pore size
of nanotubes. Carbon nanotubes (CNTs) are nanomaterials known for its excep-
tional properties, like high surface area and adsorption capacity. Due to these unique
properties, the potential use of CNTs in heavy metal contaminated water or soil has
been studied. Single-walled carbon nanotubes (SWCNTs) and multi-walled carbon

nanotubes (MWCNTs) have been successfully used for the extraction of Zn from
aqueous media (Lu and Chiu 2006). Organic contaminants such as trihalomethanes
and 1, 2-dichlorobenzene have been removed with the aid of carbon nanotubes
(Peng et al. 2003; Kondratyuk and Yates 2005; Lu et al. 2005).
8.3.6 Dendrimers and Nanosponges
These are the extremely branched polymers having a particular composition. These
types of structures are specially designed so as to capture the metal ions and inhibit
them to dissolve in other media. Poly amidoamine dendrimers are the dendrimers
which have been used for the extraction of Cu(II) from aqueous media (Diallo et al.
1999). Various dendrimers can be cross-linked with each other in order to capture
the large sized organic contaminants. Poly (amidoamine) dendrimers containing
different functional groups were used for the removal of copper from contaminated
soil. Effects of dendrimer dose, generation number, pH, terminal functional groups,
and ionic strength on the removal efficiency were investigated through a series of
column tests. About 90% of copper was removed from soil under test, with the use
of 66 bed volumes of 0.10% (w/w) of a generation 4.5 dendrimer with carboxylate
terminal groups at pH 6.0 (Xu and Zhao 2005). The dendrimers can be used as a
reusable and high-capacity extracting agents for removal of heavy metals from con-
taminated soils. In a research conducted by Arkas et al. 2006, hybrid filters were
prepared with dendritic and cyclodextrin nanosponges. These filters were then
investigated for their cleaning potential of polluted water and it was reported that
more than 95% of aromatic hydrocarbons were successfully removed from water.
8.3.7 Magnetic Nanoparticles
These are small sized magnetic particle which are further divide into 4 types depend-
ing upon their interaction with magnetic field i.e. paramagnetic, ferromagnetic, fer-
romagnetic, anti-ferromagnetic. In recent years, there has been an increase in the
use of engineered magnetic nanoparticles for remediation and water treatments. The
most popular magnetic nanoparticles are iron-based nanoparticles, Fe3O4, nZVI and
g-Fe2O3. Although, these nanoparticles are iron-based but, their chemical proper-
ties, reactivity and capability for pollutant removal is different from other iron based
nanoparticles. These magnetic nanoparticles have a large removal capacity, fast
kinetics and high reactivity for contaminant removal due to their extremely small
particle size and high surface area to volume ratio. Various reports are available for
the removal of contaminants from aqueous media with the addition of magnetic
nanoparticles. Different types of magnetic nano materials have been used for extrac-
tion of organic (dyes) and inorganic (heavy metals) pollutants.
8.5 Nanobioremediation (NBR) 205
8.4 Green Synthesis of Nanoparticles for Soil Remediation
Various physical and chemical methods have been used for production of nanopar-
ticles, but these processes involved the use of various toxic chemicals which impose
detrimental effects on the environment and living being. In order to develop effec-
tive and ecofriendly process for the production of nanoparticles, various biotic
sources comprise of Plants, Bacteria, Yeast and Fungi (Table 8.3 represents the list
of plants, bacteria, yeast and fungi) have been used. This production of nanoparti-
cles from the biotic sources is termed as green synthesis. Figure 8.2 shows a gener-
alized procedure for the synthesis of nanopartilcles. Nanoparticles from plant
sources are more important than other sources because of their easy handling and
availability. Moreover, in depth research is required in order to elaborate the mecha-
nism of action of nanoparticles as well as their effect on plant gene expression.
Bacteria have been most commonly used for the production of iron nanoparti-
cles. In a study by Bharde et al. 2005 Actinobacter sp. was used to produce spherical
iron nanoparticles under aerobic environment. Another study was conducted to pro-
duce greigite (Fe3S4) and maghemite (γ-Fe2O3) nanoparticles with the same bacte-
rial spp. Thermophilic strain Thermoanaerobacter has also been used for the
production of magnetite nanoparticles. In a study by Machado et al. 2013, green
nZVIs have been synthesized using extracts of vine leaves, grape marc and black tea
and it has been found that these green nanoparticles successfully degrade the ibu-
profen (as a contaminant) in aqueous media. These results indicated that green
Nanobioremediation can be a promising alternative to traditional remediation
technologies.
8.5 Nanobioremediation (NBR)
Among the innovative methods of in situ remediation, the nanotechnology (use of

nanomaterials) and its amalgamation with bioremediation and allied remediation
technologies has been the focus of various researches at laboratory and field levels
in North America and Europe (NANOREM 2013). The demands for innovative
technologies to efficiently accelerate the decontamination of contaminated sites are
increasing. Nanobioremediation is one such promising methodology which has
received great attention recently. Elemental/zero-valent metals such as iron, nickel
and palladium are being used at sites contaminated with various toxic substances to
stabilize transitional (Cr, As) and dehalogenation of persistent organic compounds
(Murugesan et al. 2011; Thome et al. 2015). The success of nanotechnology in the
field of remediation is because of small size (<100 nm) of nanoparticles, large sur-
face area and unique chemical characteristics (Tosco et al. 2014). Recalcitrant
organic compounds such as tetrachloroethylene (TCE) and polyvinyl biphenyls
(PCBs) can be effectively degraded through nanoremediation compared to microre-
mediation. Among all, the use of nanoiron (nZVI) is most effective due to its low
Table 8.3 List of various plants, bacteria, yeast and fungi used for the synthesis of nanoparticles
Nanoparticle Plant Bacteria Yeast and fungi References
Silicon-geranium Stauroneis sp. – – Ali et al. (2011)
Gold and silver Emblica officinalis, Citrus – Verticillium sp. and Shankar et al. (2003a, b, 2004), Ankamwar et al. (2005), Song et al.
sinensis, Coriandrum sativum, Fusarium oxysporum (2008), Narayan and Sakthivel (2008), Song et al. (2009), Kasthuri
Diopyros kaki (persimmon), et al. (2009), Philip (2009), Popescu et al. (2010), and Samim et al.
Hibiscus rosa sinensis, (2011)
Pelargonium graveolens,
Phyllanthium
Silver Nerium indicum, Elettaria Bacillus cereus, B. Yeast strains MKY3, Klaus et al. (1999), Joerger et al. (2000), Klaus et al. (2001), Shankar
cardamomom, Parthenium thuringiensis, B. Coriolus versicolor, et al. (2003a, b), Ahmad et al. (2003), Kowshik et al. (2003), Zhang
hysterophorus, Ficus indica, subtilis, Oscillatory Phaenerochaete et al. (2005), Chandran et al. (2006), Mohanpuria et al. (2008), Li et al.
Azadirachta indica, willei NTDMO1, chrysosporium, (2007), Vigneshwaran et al. (2007a), Vigneshwaran et al. (2007b),
Pelargoneum graveolens, Pseudomonis stuzeri, Cladosporium Basavaraja et al. (2008), Gade et al. (2008), Parashar et al. (2009),
Desmodium triflorum, Ocimum Lactobacillus strains, cladosporioides, Gurunathan et al. (2009), Balaji et al. (2009), Sanghi and Verma
sp., Brassica juncea, Corynebacterium, Aspergillus flavus, (2009), Shekhawat and Arya (2009), Jain et al. (2009), Ingle et al.
Pterocarpus santalinus, Escherichia coli, Fusarium semitectum, (2009), Nithya and Ragunathan (2009), Raut Rajesh et al. (2009),
Pongamia pinnata, Ureibacillus F. oxysporum, Saifuddin et al. (2009), Pugazhenthiran et al.(2009), Ganesh and
Clerodendrum inerme, Rumex thermosphaericus Extremophillic yeast, Gunasekaran (2009), Sintubin et al. (2009), Nanda and Saravanan
hymenosepalus, Euphorbia Staphylococcus aureus Pleurotus sajor-caju, (2009), Tripathy et al. (2010), Binupriya et al. (2009), Thakkar et al.
hirta, Gliricidia sepium, Carica Aspergillus niger, A. (2010), Philip (2010), Thirumurugan et al. (2010), Raut et al. (2010),
papaya (fruit), Aloe vera oryzae, Trichoderma Farooqui et al. (2010), Gade et al. (2010), Sathyavathi et al. (2010),
extract, Coriandrum sativum, viride, Fusarium solani, Mallikarjuna et al. (2011), Priya et al. (2011), Ahmad et al. (2011),
Capsicum annum, Avicennia Mourato et al. (2011), Mubarak et al. (2011), Jain et al. (2010), Juibari
marina, Rhizophora mucronata, et al. (2011), Selvi and Sivakumar (2012), Sunkar and Nachiyar (2012),
Ceriops tagal, Sonchus asper, GnanaJobitha et al. (2012), George et al. (2012), Gnanadesigan et al.
Opuntia, (2012), Jaganathan et al. (2012), Dhas et al. (2013), Ericka et al.
(2013), Kasi et al. (2013), and Verma et al. (2013)
Gold Medicago sativa, Syzygium Alkalothermophilic – Gardea-Torresday et al. (2002), Sastry et al. (2003), Ankamwar et al.
aromaticum L, Chenopodium actinomycete (2005), Smitha et al. (2009), Song et al. (2009), Sintubin et al. (2009),
album L., banana peel, Camellia Thermomonospora sp., Ankamwar (2010), Bankar et al. (2010), Arul Kumar and Sabesan
sinensis L., Magnolia kobus, Pseudomonas (2010), Vankar and Bajpai (2010), Deshpande et al. (2010),
Mentha piperita L., Azadirachta aeruginosa, Thirumurugan et al. (2010), Dwivedi and Gopal (2011), Ali et al.
indica, Justicia gendarussa L., lactobacillus strain (2011), Parida et al. (2011), Boruah et al. (2012), Fazaludeena et al.
Allium cepa L., Macrotyloma (2017), Aromal et al. (2012), and Das et al. (2012)
uniflorum, Mirabilis jalapa L.,
Terminalia catappa,
Cinnamomum zeylanicum,
Dyopiros kaki., Terminalia
catappa L., Amaranthus
spinosus, Mucuna pruriens,
Silver, nickel, Medicago sativa, Brassica – – Bali et al. (2006)
cobalt, zinc and juncea and Helianthus annuus
copper
Platinum Ocimum sanctum L., Diopyros – – Song et al. (2010), and Soundarrajan et al. (2012)
kaki
Palladium Gardenia jasminoides Ellis., Desulfovibrio – Nair and Pradeep (2002), Jia et al. (2009) Sathish et al. (2009), Xin
Cinnamomum zeylanicum desulfuricans NCIMB et al. (2010), and Petla et al. (2012)
Blume., Glycine Max L., 8307
Cinnamomum camphora L.
Lead Vitus vinifera L., Jatropha – – Joglekar et al. (2011), and Pavani et al. (2012)
curcas L.
Magnetic Aloe vera Magnetosirillium – Gericke and Pinches (2006), Mohanpuria et al. (2008), and Phumying
magneticum, Sulphate et al. (2013)
reducing bacteria
CdS – Clostridicum Candida glabrata, Dameron et al. (1989), Sweeney et al. (2004), and Mandal et al. (2006)
thermoaceticum, Schizosaccharomyce
Klebsiella aerogens, pombe
Escherichia coli
(continued)
Gold nanowires – Rhodopseudomonas – He et al. (2008)
capsulate
As-S nanotubes – Shewanella sp – Jiang et al. (2009)
ZnS – Sulphate reducing – Mandal et al. (2006), Labrenz et al. (2000), Rathore et al. (2008),
bacteria Murugadoss et al. (2009), and Sharma et al. (2009)
PbS – – Torilopsis spp., Kowshik et al. (2003), and Seshadri et al. (2011)
Rhodospiridium
dibovatum
CdS quantum dots – – Schizosacharomyces Kowshik et al. (2003)
pombe
Stable silver – – Aspergillus flavus, A. A. Bhainsa and D’souza (2006), and Khalil (2013)
terreus, furnigatus, A.
nidulans
Bioactive – – Usneea longissima Shahi and Patra (2003)
8.5 Nanobioremediation (NBR) 209
Fig. 8.2 Schematic representation of the generalized procedure for the biosynthesis of
nanoparticles
toxicity and cost-effectiveness. There are several research reports available wherein
zero-valent iron and nanosize Feo have been used as reducing agent to remediate
sites contaminated with halogenated compound, polychlorinated biphenyls, herbi-
cides, pesticides, aromatic polycyclic hydrocarbons and metals (Ponder et al. 2000;
Zhang et al. 2005; Devor et al. 2006; Elliott et al. 2009; Sakulchaicharoen et al.
2010). A problem associated with the application of nanoparticles in soil remedia-
tion is the clogging of soil pores (Reddy 2010). Although, with the use of surface
coating [sodium carboxymethyl cellulose (CMC), guar gum, lactate etc.], the stabil-
ity and reactivity of the nanoparticles can be prolonged (Wei et al. 2010;
Jiemvarangkul et al. 2011). Another limitation is that nanoparticles have been
reported to have toxic effects on some microorganisms eg. E. coli, Staphylococcus
aureus, Dehalococcoides spp. (Li and Jiang 2009; Gordon and Margel 2011). There
are several reports on the integrated use of nanoparticles and bioremediation. Singh
et al. (2013), succeeded (99% efficiency) in remediation of soil contaminated with
lindane, in 6 days, by the combined action of CMC-Pd/Fe nanoparticles and
Sphingomonas species. Le et al. (2015) reported the degradation of 89% degrada-
tion of Aroclor 1248 (PCB) using nZVI followed by the 90% biodegradation of the
products Burkhalderia xenovorans.
Fig. 8.3 Nanobioremediation of polluted soil using injection method and incorporation of nZVI
on target site
Nowadays stabilized ZVI nanoparticles are most commonly used for the extrac-
tion of organic and inorganic contaminants from polluted soil, sludges and ground-
water (Tratnyek et al. 2011). Figure 8.3 represents the schematic diagram of
incorporation of nZVI particles in the deep soil (through injection well) and over the
soil surface for remediation of contaminated soil. Different types of organic coat-
ings on these nanoparticles pose considerable effect on the reactivity of ZVI such as
(1) organic coating can modify the surface of ZVI and increase the rate of adsorp-
tion of contaminants over the nanoparticle’s surface, (2) these coatings can act as a
catalyst through shuttling electrons, which speed up the redox reactions, and (3)
coatings of different organic polyelectrolytes increases the mobility of hydrophobic
contaminants. Combination of ZVI with metal catalysts (e.g. Pb) has been shown to
be one of the most efficient and promising nanoparticle for effective degradation of
chlorinated contaminants (Urbano and Marinas 2001). Figure 8.4 represents the
mechanism of reductive dechlorination by nZVI of its surface with the aid of Pb as
a catalyst. This Fe-Pb bimetal system enhances the removal efficiency of nanopar-
ticle and do not allow the accumulation of harmful byproducts. The reactions over
the surface of Fe-ZVI include adsorption of H2 over the surface of metal, production
of reactive hydrogen species which further lead to the dechlorination reaction.
8.6 Conclusions 211
Fig. 8.4 Schematic representation of reductive dechlorination mechanism performed by nZVI,

with or without catalyst (Pb)
8.6 Conclusions
Nanotechnology holds a promising potential for environmental protection as well as

soil remediation. These particles degrade the contaminants to a low toxic level and
enhance the rate of biodegradation, thus it is an efficient, sustainable and effective
approach to remediate the polluted sites of different regions. This technology can
provide an efficient and ecofriendly alternative to other remediation method for
extraction of various organic (PAHs) and inorganic (heavy metals) pollutants from
soil. Distinctive physical and chemical properties of nanoparticles have made them
the particles of choice for restoration of the polluted environments. Nanoparticles
obtained from hyper accumulator plants, bacteria, yeast and fungi are of great
importance for remediation of polluted soil. Numbers of researches are now focused
on the green synthesis of nanoparticles i.e. the production of nanoparticles using
biological materials such as plants, bacteria, algae and fungi. Furthermore, research
is required so as to achieve the sustainability of nanoparticles and to understand the
detailed mechanism of their mode of action.
In the coming years, further modification and adaptation should be performed
in the field of nanotechnology that will surely enhance the efficiency of
Nanobioremediation. Likewise, detailed study regarding the effect of nanoparti-
cles on soil flora should be recommended. It is crucial to design a robust protocol
that may have least impact on soil microbes, so as to maintain the soil
biodiversity.
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Chapter 9
Case Studies and Future Prospects of Soil
Remediation Strategies
Abstract Soil remediation strategies are subjected to the quality and quantity of the
contaminant(s) as well as geographical conditions of the target site. There is no
single soil remediation method that can aid as a ‘silver bullet’ to restore the environ-
mental deterioration without any residual effect. For successful soil remediation
different physical, chemical and biological strategies can be implemented in an inte-
grated way. This chapter encompasses various case studies related to the implemen-
tation of remediation strategies on a large scale. This chapter also focuses on the
explicit information and recent advances on the available soil treatment techniques
and their future prospects. Thus, the ideal soil remediation strategies ensure envi-
ronmental protection by using natural resources for sustainable soil remediation
which is economically and environmentally beneficial for the society.
Keywords Soil contamination · Soil washing · Windrows · Bioventing · Microbial

degradation · Chemical oxidation
9.1 Introduction
Remediation of soil polluted with various organic and inorganic contaminants can
take place under in situ and ex situ conditions through physical, chemical and bio-
logical treatments. For a successful remediation strategy, in depth understanding of
physical and chemical properties of pollutants as well as that of the soil is required.
Great care should be taken while choosing a remediation strategy for soil contami-
nated with a particular pollutant. Various treatments have been performed at large
scale for the remediation of polluted soils through physical, chemical and biological
methods. Established case studies reveal that the researches which were conducted
at laboratory levels can be successfully implemented on a large scale. For e.g. a soil
treatment was conducted by Dunton Environmental, Birmingham, UK in 1979 to
remediate the soil polluted with tarry liquid and sludges of gas work wastes in
Saltisford Gasworks and Fire Station Warwick, UK. Bio-accelerators were devel-
oped to enhance the metabolism of naturally occurring microbes, which success-
fully degraded the soil contaminants. The concentrations of pollutants were reported

222 9 Case Studies and Future Prospects of Soil Remediation Strategies
to be decreased by 83% within 3 weeks of treatment. In this way, immobilization

and removal of pollutants was done with the aid of microbe-mediated remediation.
A study was conducted in Rovinari and Copsa-Mica, the two heavily polluted areas
of Romania. The soil was highly contaminated with heavy metals like copper, arse-
nic, cadmium, lead and mercury. Several plants spp. (Camelina sativa, Bunias ori-
entalis, Thlaspicae rulescens) were planted on the contaminated soil for
phytoextraction and phytostabilization of the contaminants. Among this plant spe-
cies, C. sativa was found to be highly effective for remediation of polluted soils. This
remediation project started in 2013 and is still ongoing. Another soil treatment was
performed on Ni-contaminated industrial soil of Mysore, India. This treatment was
based upon the applications of nanoparticles. As mentioned before, nanotechnology
offers the possibility of efficient removal of pollutants. In this remediation set up
zero valent iron nanoparticles were used for removal of Ni from industrial soil. The
initial concentration of nickel in the soil was 10–18.1 mg/kg. It was observed that
with the aid of nanoparticles Ni concentration decreased upto 75–90% in the soil.
9.2 Case Studies on Soil Remediation
Following are the few case studies of soil remediation treatments that were con-
ducted in different parts of the world using physical, chemical and biological
methods:
9.2.1 Case Study 1
Location
Eastgate, Memphis, U.S.A
Oxboro, South Bloomington, North America
Source of Soil Contamination
Waste and wastewater from dry cleaning operations
Time Period
Eastgate: February 1994–November 1996
System reactivation in March 2001–June 2001
Oxboro: October 10, 1997-Not specified
Purpose of Treatment
Use of in situ SVE (Soil vapor extraction) to remediate soil and groundwater pol-
luted with chlorinated solvents at dry cleaner sites.
Contaminants
1,1-Dichloroethene (1,1 DCE), Methylene Chloride, Tetrachloroethene (PCE),
Vinyl Chloride, 1,2-Dichloroethene (1,2 DCE), Benzene, Toluene, cis-1,2-
Dichloroethene (cis-1,2 DCE), Acetone, Trichloroethene (TCE), Naphthalene.
9.2 Case Studies on Soil Remediation 223
Technology: Soil Vapour Extraction

In Situ SVE system was operated in Easgate which also involve turning off the sys-
tem for shorter time periods. Trail period of 2001 was operated 24 h per day with an
air flow rate of 300 cfm. Total 2000 lbs of granular carbon was used for the treat-
ment. In Oxoboro, SVE systems include wells for soil venting with a blower system
which was checked on weekly and monthly basis.
Results
In Eastgate approximately 1350 lbs of PCE and TCE were removed from soil during
initial time period of treatment. In treatment of 2001, 0.16 lbs of PCE and TCE were
removed from contaminated soil. In Oxoboro concentration of PCE in soil decreased
from 160 mg/m3 to 0.3 mg/m3 in October 1997. TCE were completely removed
from the contaminated soil.
9.2.2 Case Study 2
Location
Former Market Place Shopping Center Site, Hilton Head, South Carolina
Denver Colorado Dry Cleaner, Denver, Colorado
United Cleaners Site #1973, Lemont, U.S. state of Illinois
Waste and wastewater from dry cleaning operations
Time Period
Site 1- Former Market Place – June 1, 2002
Site 2- Denver Colorado – April 2001
Site 3- United Cleaner #1973 – August 26, 2002
To treat the soil contaminated with chlorinated solvents with the aid of in situ treat-
ment technologies.
Contaminants
Dichloroethene (DCE), Tetrachloroethene (PCE), Volatiles-Halogenated,
Trichloroethene (TCE)
Technology: ISCO and In Situ Heat Soil Vapor Extraction (HSVE)
Two treatments were used i.e. ISCO and In Situ Heat Soil Vapor Extraction (HSVE).
At ‘Site 1’ ISCO was performed with the aid of ozone as an oxidant. At ‘Site 2’
ISCO was performed with modified Fenton’s Reagent. Two injections events were
used in the area to treat contaminated soil. Total 26,987 gallons of oxidation reagent
were applied through temporary injection systems. At Site 3 in situ HSVE technique
was performed by applying heat to the soil, increased the volatility of contaminants,
then SVE was performed to remove the contaminants.
Results
Site 1 showed a decrease in PCE concentration from 26,800 g/L to 704 g/L. At Site
2, average decreases in total concentration of all the contaminants were reduced
from 550 g/L to 52 g/L. At Site 3, concentration of PCE in soil was reduced from
4,300,000 g/kg to 2,400,000 g/kg. The treated area was completely remediated after
the treatment.
9.2.3 Case Study 3
Location
East Multnomah County Groundwater Contamination Site, Gresham, Oregon
Suspected releases from site operations, including overflow from an underground
storage tank (UST), spills, and on-site land disposal
Time Period
June 1991 to present
Implementation of different treatments to remove the chlorinated solvents from soil.
Contaminants
Trichloroethene (TCE), Tetrachloroethene (PCE), Cis-1,2-dichloroethene (DCE),
heavy metals
Technology
Air sparging
Air sparging, Groundwater pump, soil vapor extraction. Air sparging was per-
formed in two wells which enhanced the volatilization of VOCs from groundwater.
SVE was ceased from March to October 1999 and ceased permanently in December
2001.
Results
In year 2003, total 958 pounds of VOCs were removed from the soil after 13 year of
treatment period.
9.2.4 Case Study 4
Location
Morses Pond Culvert, Wellesley, Massachusetts
Chromium-laden pigments from former paint factory
Time Period
September–October 2001
In situ treatment of chromium contaminated soils
Contaminants
Chromium, zinc, and lead
Technology: ISCO
In situ chemical reduction was performed on the target site with the use of calcium
polysulfide. Total 40 injection wells were installed with a distance of 10 ft apart
from each other and 5–25 ft in depth. First treatment was performed in western part,
then eastern part of the area. Total 56,800 gallons of calcium polysulfide was
injected into soil through the injection wells.
Results
Concentration of total chromium was decreased from 67,000 mg/kg to 24 mg/kg.
9.2.5 Case Study 5
Location
Canada
Not mentioned
Onset of treatment
July 2010
This treatment was performed for remediation of soil contaminated with
hydrocarbons.
Contaminants
Benzene, Ethylbenzene, Xylene Mixture, Toluene
Technology: Biopile or Landfarming
Biopile or landfarming treatment was performed on the polluted soil (Fig. 9.1 a, b)
This process reduced the concentration of hydrocarbons in soil through the aid of
biodegradation. The conditions like oxygen, Moisture, nutrients, heat and pH were
maintained in order to enhance the biodegradation rate. Two piles of polluted soil
with size 25m3 were formed. Piles were of 20 ft in height and the air supply was
equally distributed under the soil piles. These piles were covered with plastic paper
to control the evaporation and to enhance the solar heating. VOCs were volatilized
and entered into the air stream. This air stream was then treated to remove the con-
taminants. One pile was treated with microbial consortia and second pile was treated
Fig. 9.1 Biopile-set up (a) Biopile or Land farming of soil (b) Biopiles covered with plastic sheet
with soil reaction mixture. Both the piles were then monitored at regular intervals
for their degradation efficiency.
Results
The time period for normal remediation was about 4 weeks. While, for full scale
remediation extra 7–14 days were required. This treatment was able to remediate
approximately 650,000 cubic meters of material/year.
9.2.6 Case Study 6
Location
Somerset Energy Refining
Somerset Kentucky

Not mentioned
Onset of treatment
September 2009
To evaluate the remediation efficiency of a mixture of Soil Rx, microbial consortia
and biodegradable natural enzymes for removal of hydrocarbons from soil.
Contaminants
Crude Oil and Gasoline
Technology: Microbial Degradation
Treatment area consisted of 2 frames of sizes 4’ × 4’ polluted with crude oil and
gasoline. A mixture of 20:1 of Soil Rx and water was prepared and 2.5 gallons of
this mixture was supplied to each frame. Samples were taken from the treatment
area after Ist, 5th and 14th day. All testing was completed by Environmental Science
Corporation in Mt. Juliet Tennessee. This remediation treatment converted the pol-
lutants into biodegradable products like Co2, H2O and enzymes.
Results
In the treated area with crude oil the concentration of toluene was reduced from 54
to 1.8 mg/kg, Benzene was reduced from 15 to 0.13 mg/kg, Xylene was reduced
from 190 to 45 mg/kg and Ethylbenzene was reduced from 33 to 4.4 mg/kg. In the
area polluted with gasoline, concentration of Benzene was reduced from 140 mg/kg
to BDL (below detectable levels), Ethylbenzene was reduced from 810 to 0.64 mg/
kg, Toluene was reduced from 2000 to BDL, Xylene was reduced from 5500 to
8.4 mg/kg. All the pollutants of this area were converted into low toxic compounds
within the first five days of the treatment.
9.2.7 Case Study 7
Location
Four Mile, Bell County, Kentucky
Crude oil released during oil transfer and pick up.
Onset of treatment
June 2010
This treatment was conducted to create a cost effective method so as to eliminate the
crude oil from soil site.
Contaminants
Crude Oil
Technology: Microbial Degradation
Contaminated site was applied with a mixture of Soil Rx and water (20:1) with
drench method. 5 gallons of Rx mixture was applied in each treatment (Fig. 9.2).
Contaminated site was treated on 10 May, 31 May and 11 June 2010.
Results
The results depicted that average 59% of VOCs concentration was decreased in soil
after the 12th day of treatment. Total 68% of benzene was removed, after 12th day
of treatment. This study revealed that soil Rx mixture can be used as an effective
remediation tool for the treatment of soil contaminated with crude oils.
9.2.8 Case Study 8
Location
Loring Air Force Base Tank Farm, U.S.A
Fuel was released from large fuel tanks placed at Loring Air Force Base Tank Farm,
U.S.A
Onset of treatment
October 2010
Fig. 9.2 Soil polluted with crude oil (Four Mile, Bell co., Kentucky)
Main purpose of this treatment was to remediate the polluted soil by using eco-
friendly and inexpensive method and to develop new advances in anaerobic biore-
mediation of soil polluted with hydrocarbons.
Contaminants
Fuel
Technology: Bioventing
Initially two bioventing remediation treatments were performed on the polluted soil
in 1996 to 2005. These treatments were not proved successful. After that treatment,
Soil Rx mixture was injected into the polluted soil through boring holes on 4th
October 2010 (Fig. 9.3). Samples were taken from the holes on 28th June 2011 foe
evaluation and the entire site was retreated with the same mixture.
Results
Although application of soil mixture is a suitable remediation tool. But, the result
obtained with the above treatment was not so effective for soil remediation.
9.2.9 Case Study 9
Location
Nynas UK AB, Bitumen Refinery, Dunde, Scotland
Oil soil polluted with hydrocarbons
Time Period
2 years 3 months, 2013-2015
Fig. 9.3 Overview of the target area (C) Control (T) Treated
To develop a sustainable and ecofriendly environmental solution to treat the old soil
polluted with hydrocarbons.
Contaminants
Hydrocarbons
Technology: Windrows
Polluted soil was screened so as to remove the large sized material from it. Soil was
then formed into windrows and covered with breathable material i.e. fleece (Fig. 9.4
a–d). After that soil was implemented with hydrocarbon degrading bacteria, air sup-
ply and sufficient amount of nutrients. Nutrients level, air supply and other condi-
tions were regularly monitored throughout the treatment period.
Hydrogen degrading bacteria were responsible for the degradation of
hydrocarbons.
Results
The above bioremediation treatment was proved as an efficient remediation method.
Total 72% of hydrocarbons were removed from the soil.
Fig. 9.4 Soil remediation: (a) Remediation area at the start of work (b) remediation of windrows
(c) turning of windrows (d) Remediated soil
9.2.10 Case Study 10
Location
Not mentioned
Not mentioned
Time Period
8 months
To remediate the soil polluted with heterogeneous organic compounds (BTEX,
PAH, TPH) ranging from 1000 mg/kg to 10,000 mg/kg. Total 2700 m3 of soil was
polluted with hydrocarbons.
Contaminants
Heterogeneous organic compounds i.e. BTEX, PAH, TPH
Technology: Chemical Oxidation
Initial sample was taken from the polluted soil during the period of licensing and
other formalities. A laboratory set up was made near the polluted soil to evaluate the
pH and other conditions of the soil. Fixed amount of Klozur Persulphate and Lime
was then added into the soil with the help of mixing head (Fig. 9.5a–b). These
chemicals served as oxidizing agents and oxidized the pollutants into low toxic
compounds. Additional water was added into the soil in order to enhance the pro-
cess of oxidation. Soil was analyzed periodically throughout the treatment.
Results
The soil was remediated efficiently within 8 weeks of the treatment.
Location
Former filling station site, South Glasgow, U.K
Not mentioned
Time Period
1 month
To remediate the contaminated soil and water polluted with hydrocarbons.
Contaminants
Hydrocarbons
Fig. 9.5 Soil remediation: (a) Addition of chemicals into polluted soil (b) Soil mixing
Technology: Chemical Oxidation

Remediation treatment was started with one initial round of vacuum extraction of
contaminants from the polluted soil. After that, ISCO was performed with a chemi-
cal oxidant (Chemox) over the soil surface (Fig. 9.6 a–d). Chemox was applied in
the soil through Direct Push Injection. Total three treatments were applied on soil
with the aid of chemox. It served as oxidizing agents and oxidized the pollutants
into low toxic compounds.
Results
Two treatments proved sufficient for soil remediation.
Fig. 9.6 In situ remediation of polluted soil (a-d) through chemical oxidation
Location
Olympic Park, Stratford, London
Not mentioned
Time Period
July 2007, 78 weeks
To remediate the polluted soil of Olympic park
Contaminants
Hydrocarbons (TPH, Benzopyrene,), Heavy metals (As, Cu, Pb,)
Technology: Soil Washing
For soil remediation of above polluted soil, soil washing method was used. This
treatment was performed by installing two soil washing units at two different sites
of the target soil (Fig. 9.7a, b). These plants were fitted with washers, scrubbers,
separators, hydrocyclones, drum magnets and recovery systems.
Fig. 9.7 Soil remediation (a, b) using soil washing method
Results
Soil washing method proved beneficial for the removal of Hydrocarbons (TPH,
Benzopyrene) and Heavy metals (As, Cu, Pb,) from the target soil.
Location
Cape Canaveral Air Force Station, U.S.A
Not mentioned
Time Period
July 2002–January 2003.
To remediate the soil contaminated with TCE.
Contaminants
TCE
Technology: EZVI
Different injection methods for EZVI were implemented on the contaminated soil.
Soil samples were taken at different time periods i.e. before the treatment, during
the treatment and after 6 weeks of the treatment. The whole project was funded by
NASA and Department of Defense Environmental Security Technology Certification
Program (ESTCP ER-0431).
Results
Total 50% of TCE was effectively removed from the soil.
Location
Naval Air Engineering Station, Lakehurst, New Jersey, U.S.A
Not mentioned
Time Period
Not mentioned
To remediated the polluted site of Aircraft Launch facility
Contaminants
PCE (Perchloroethylene), TCE (Trichloroethylene), TCA (Trichloroacetic acid),
c-DCE (cis-dichloroethene), VC (Vinyl chloride)
Technology
BNP (boronate nanoparticle)
Total 300 lbs of BNP nanoparticles were prepared in form of slurry. Total 18,000
gallons of water was mixed with 300 lbs of BNP. This slurry was then injected using
DPT (direct push technology) at 10 different locations of Northern plume and at 51
locations of Southern plume. In one area the injections were operated in a linear
pattern and in a grid pattern for the other area.
Results
The average decrease of contaminants was 74%. Two major contaminants i.e. TCE
and DCE showed an average decrease of 79% and 83% respectively.
Location
Not mentioned
Accidental release of gasoline
Time Period
1999–2000
To evaluate the remediation potential of plant species grown on gasoline contami-
nated area.
Contaminants
Gasoline
Technology
Phytoremediation
The plant species which were used included, willows, cattail, cottonwoods,
arrowhead and bulrush. Cottonwood and Black willow were grown near the edge of
drainage area.
Results
At the end of year 2000, these plants were evaluated in terms of their phytoremedia-
tion potential. It has reported after 3 growing seasons, total 90% of the contaminant
was removed from the target site.
Location
CEA research centre, Grenoble, France (Fig. 9.8)
Radioactive solid waste released from research station
Time Period
2006–2013
To achieve the radioactivity level below 0.04 Bq/g for α emitters and below 0.4 Bq/g
for β/γ emitters.
Contaminants
U-234, C-14, Cs-137, Sr-90, U-238 and Co-60.
9.3 Future Prospects 237
Fig. 9.8 Grenoble, France, CEA research centre
Technology
18 drillings (2–3 m in depth) were set inside the area with radioactivity levels above
2 Bq/g and 37 drillings were implemented on other areas. A gamma detector was
placed with each drilling. Samples were taken from each drilling at regular inter-
vals. These samples were then analysed chemically and radio logically in the
laboratory
Results
Soil remediation was started in 2009 and the soil was partially remediated by the
beginning of the year 2013.
9.3 Future Prospects
The remediation technologies that have been discussed are need commercialization
and field applicability in the underdeveloped countries where urbanization, industri-
alization, and agriculture are leaving a legacy of ecosystem pollution. There is no
single method that can aid as a ‘silver bullet’ to restore the polluted environments
(Azubuike et al. 2016). For a remediation technique to become acceptable, the pre-
dictability and effectiveness of remediation should be illustrative (Kuiper et al.
2004). The restoration and sustenance of heavy metal contaminated sites require
thorough inspection followed by the application of most suitable remediation strat-

egy. Various parameters should be taken into consideration to design eco-friendly,
most effective and quick remediation technique. These parameters include soil
structure, pollutant nature, microbial composition and hydrogeology (movement of
pollutants) (Dua et al. 2002; Kuiper et al. 2004). The molecular biomonitoring
methods can be applied for evaluating the intrinsic biodegradation potential, observe
the enhanced bioremediation, and estimating the success of bioremediation. Besides
this, background knowledge of sources, chemistry and risk associated with toxic
metals, and in depth understanding of molecular, cellular and biochemical processes
which are affected by the heavy metals can enhance the selection of an appropriate
remediation strategy.
Phytoremediation is an emerging, non-destructive, aesthetically pleasing and
low-cost technique which uses the potential of green plants to metabolize various
contaminants (heavy metals, chlorinated solvents, PAHs, explosives, surfactants,
pesticides, radionuclides) from the ecosystem, in their tissues. However, this tech-
nology is still in its infancy and full-scale application is needed. Mostly, all the
hyperaccumulator plants are of small size hence genetic modifications can be done
for these species to increase the biomass of hyperaccumulators so as to produce
efficient phytoremediators. The biggest obstruction to the progression of phytore-
mediation may be public opposition to genetic modification. However, there are still
some fundamental researches required for complete understanding of the phytore-
mediation mechanism and to develop new strategies to accelerate the rate of
phytoremediation.
In North America, a couple of new technologies have been adopted for remedia-
tion of PCE and TCE from polluted soil. The in situ use of zerovalent iron (ZVI)
remediated 25,000 PPB of TCE and PCE in the ground water, brought down to 0
PPB within a span of 6 years. At the sites where contamination from mines gets
leached into the groundwater, permanganate products have been used which aggres-
sively oxidize the TCE and PCE in ground water. In Canada, an innovative treat-in-
place (in situ) method of bio-remediation uses superbugs which are injected into the
soil. One such microorganism that has attracted the attention of geneticists is
Deinococcus radiodurans, a soil bacterium which can bear radiations several times
higher than humans. The bacterium has the potential to break down radioactive Hg
and toluene. After being engineered to work in radioactive waste environments, it
can be used to decompose wastes further in the fields of medicine, energy and mili-
tary. Molecular bio monitoring for bacteria and their genes responsible for degrada-
tion are of great importance in order to understand the molecular phenomenon
behind remediation processes. It could be applied for several purposes within the
environmental biotechnology: to map the genetic constitution of the microorgan-
isms, to diagnose the polluted soil, to check the remediation potential, to increase,
estimate and control the remediation process. In the long run, soil remediation by
using natural resources shall ensure sustainable development, which will prove
environmentally and economically safe for the people. Symbiotic association of
bacteria and plants is beneficial for the degradation of certain recalcitrant com-
pounds. Besides that, there is only some information available on plasmid dynamics
References 239
and the characterization of plasmids from natural resources. Multidisciplinary

approaches with a blend of engineering-based and bioremediation strategies might
be the most reliable solution (Kuiper et al. 2004; Pilon-Smits 2005). Alternatively,
multiple bioremediation approaches can be used in combinations, to overcome the
limitations including the cost of the technique(s) (Cassidy et al. 2015; García-
Delgado et al. 2015; Martínez-Pascual et al. 2015).
For sure there is promising future for microbial biodegradation techniques for
treating the contaminated soils. Basically, this technique is less expensive and
requires low start-up. The aforementioned microorganisms may either be effec-
tively isolated from their regular habitat or procured from commercial suppliers.
The soil metagenomics approach (next generation sequencing and bioinformatics
analysis) can assist to efficiently monitor the clean-up of contaminated environ-
ments and develop successful strategies to ameliorate the effect of pollutants on the
ecosystem(s) (Bouhajja et al. 2016). Bioelectrochemical technology (microbial fuel
cells: MFCs) has shown a promising potential for remediation of soil, sludge and
sediments contaminated with organic and inorganic pollutants (Adelaja et al. 2014;
Li et al. 2017). In the last couple of years, there have been great advances which
have opened new vistas for the improvement of molecular techniques to screen and
control microbial groups for expanding the current phytoremediation frameworks.
The application of nanoparticles has gained momentum in metal remediation tech-
nology because of their excellent adsorption and mechanical properties, high chem-
ical stability and unique electrical property (Singh and Prasad 2015).
It is impractical to re-establish all the characteristic features of the contaminated
soils and not each site can ever be totally recouped to a pristine state. Hence, the
application of green remediation approaches must be enough to manage the con-
taminated soils. At the same time, the efficiency, efficacy and the demand of each
technique varies with the nature of contaminant, site of contamination, feasibility of
application and the extent of human intervention. This review expects to educate
and motivate research for utilization of phytotechnologies in the coming years. In a
nutshell, bioremediation techniques are more reliable, more productive and consid-
erably less expensive than the chemical or physical strategies. Thus, future research
can incorporate various combinations of hyperaccumulator plant species and genet-
ically engineered microbes for a better remediation of contaminated soils (Azubuike
et al. 2016; Marchand 2017).
References
Adelaja O, Keshavarz T, Kyazze G (2014) Enhanced biodegradation of phenanthrene using differ-

ent inoculum types in a microbial fuel cell. Eng Life Sci 14(2):218–228
Azubuike CC, Chikere CB, Okpokwasili GC (2016) Bioremediation techniques-classification
based on site of application: principles, advantages, limitations and prospects. World
J Microbiol Biotechnol 32(11):180
Bouhajja E, Agathos SN, George IF (2016) Metagenomics: probing pollutant fate in natural and
engineered ecosystems. Biotechnol Adv 34(8):1413–1426
Cassidy DP, Srivastava VJ, Dombrowski FJ, Lingle JW (2015) Combining in situ chemical oxida-
tion, stabilization, and anaerobic bioremediation in a single application to reduce contaminant
mass and leachability in soil. J Hazard Mater 297:347–355
Dua M, Singh A, Sethunathan N, Johri AK (2002) Biotechnology and bioremediation: successes
and limitations. Appl Microbiol Biotechnol 59:143–152
García-Delgado C, Alfaro-Barta I, Eymar E (2015) Combination of biochar amendment and
mycoremediation for polycyclic aromatic hydrocarbons immobilization and biodegradation in
creosote-contaminated soil. J Hazard Mater 285:259–266
Kuiper I, Lagendijk EL, Bloemberg GV, Lugtenberg BJ (2004) Rhizoremediation: a beneficial
plant-microbe interaction. Mol Plant-Microbe Int 17(1):6–15
Li X, Wang X, Weng L, Zhou Q, Li Y (2017) Microbial fuel cell for organic contaminated soil
remedial application: a review. Energy Technol 5(8):1156–1164
elements. Doctoral dissertation, Department of Biology and Environmental Science, Linnaeus
University, Kalmar
Martínez-Pascual E, Grotenhuis T, Solanas AM, Viñas M (2015) Coupling chemical oxidation and
biostimulation: effects on the natural attenuation capacity and resilience of the native microbial
community in alkylbenzene-polluted soil. J Hazard Mater 300:135–143
Pilon-Smits E (2005) Phytoremediation. Annu Rev Plant Biol 56:15–39
Singh A, Prasad SM (2015) Remediation of heavy metal contaminated ecosystem: an overview on
technology advancement. Int J Environ Sci Technol 12(1):353–366

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Bhupendra Koul · Pooja Taak

ISBN 978-981-13-2419-2 ISBN 978-981-13-2420-8 (eBook)

Library of Congress Control Number: 2018958474

© Springer Nature Singapore Pte Ltd. 2018

prospects. Thus, the soil remediation strategies ensure environmental protection by

chemical leaching, chemical oxidation, chemical fixation, and electrokinetic

Chapter 9 Case Studies and Future Prospects of Soil Remediation Strategies

Phagwara, Punjab, India Bhupendra Koul

1 Soil Pollution: Causes and Consequences������������������������������������������������ 1

3.2.4 Froth Flotation���������������������������������������������������������������������� 64

7.4 Genetic Engineering Approaches for Phytoremediation�������������������� 165

Dr. Bhupendra Koul is serving as an Assistant Professor in the School of

ACC deaminase 1-Aminocyclopropane-1-carboxylic acid deaminase

IBI International biochar initiative

YCF1 Cadmium factor protein

Keywords Pollution · Toxic effects · Heavy metals · PAHs

In the coming years, sustainable plant productivity and availability of bio-resources

© Springer Nature Singapore Pte Ltd. 2018 1

Fig. 1.1 Types of pollution

depletion of biodiversity. Urbanization, modern agricultural practices, industrializa-

1.2 Causes of Soil Pollution

1.2.1 Natural Sources

1.2.1.1 Volcanic Eruptions

1.2.1.3 Alterations in Rainfall Patterns

1.2.1.4 Geographical Changes

1.2.2 Anthropogenic Sources

1.2.2.1 Solid Wastes

Municipal Solid Wastes (MSW)

Hospital wastes are produced as a result of diagnosis, treatment and immunization

1.2.2.2 Agricultural Practices

Causes of soil pollution

Volcanic eruptions Solid wastes

Tsunamis Construction activities

Fig. 1.3 Causes of soil pollution

Agrochemicals Aniline derivatives

Fig. 1.4 Agrochemicals responsible for soil and water pollution

2-methyl-4,6-dichlorophenoxyacetic acid. These herbicides also contain the

Fig. 1.5 Consumption pattern of pesticides in the world

Fig. 1.8 Structures of (a)

(A) (B) (C) (D)

(A) (B) (C)

tence and toxicity, while higher insecticidal activity. Bioresmethrin, fenvalerate,

(A) (B) (C)

Fig. 1.14 Structures of (a) Captan and (b) Benzimidazole

(A) (B) (C)

(D) (E) (F)

(A) (B) (C)

(D) (E) (F)

(G) (H) (I)

1.2.2.3 Radioactive Wastes

1.2.2.4 Chemical Wastes

1.2.2.5 Mining and Smelting

1.3 Harmful Effects of Soil Pollution

effect hormonal balance, membrane structure and permeability.

On plants: Abnormal germination and hypertrophy of the roots and

decreased protein synthesis, withering and drying of leaves and

Box 1.1: Quality Control of Export and Import Consignment

Box 1.2: Rejection of Export Consignment: Indian Case Study

Table 1 Notifications raised by the EU in RASFF Portal (2005–2017)

Box 1.2: (continued)

Although shallow water table enhances the changes of contamination of the

1.4 Remedies to Cope with Soil Pollution

Healthy land is an essential element in order to sustain human development and to

Box 1.3: “Swach Bharat Abhiyaan”

Phagwara, Punjab, India Bhupendra Koul

1 Soil Pollution: Causes and Consequences�� 1

3.2.4 Froth Flotation�� 64

7.4 Genetic Engineering Approaches for Phytoremediation�� 165

1.2 Causes of Soil Pollution

1.2.1 Natural Sources

1.2.1.1 Volcanic Eruptions

1.2.1.3 Alterations in Rainfall Patterns

1.2.1.4 Geographical Changes

1.2.2 Anthropogenic Sources

1.2.2.1 Solid Wastes

1.2.2.2 Agricultural Practices

1.2.2.3 Radioactive Wastes

1.2.2.4 Chemical Wastes

1.2.2.5 Mining and Smelting

1.3 Harmful Effects of Soil Pollution

1.4 Remedies to Cope with Soil Pollution

2.2 Land Farming

2.6 Soil Washing

2.9 Ion Exchange

pollutant-cleaning efficiency, no CO2 production, effective reduction in weight and

2.11 Ultrasonic Technology

3.2 Physical Methods of Soil Remediation

3.2.1 Physical Separation

3.2.2 Soil Flushing

3.2.4 Froth Flotation

3.2.5 Thermal Remediation

3.2.5.1 Electrical Resistance Heating (ERH)

3.2.5.2 Steam Injection and Extraction

3.2.5.3 Conductive Heating

3.2.5.4 Radio-Frequency Heating (RFH)

3.2.5.5 In situ Vitrification (ISV)

4.2 Chemical Leaching

4.3 Chemical Fixation

4.4 Chemical Oxidation

4.5 Electrokinetic Remediation