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Abstract Environment protection and sustainability are harmonious and sustain-
ability can be achieved by protecting our natural resources. This chapter presents
an overview of the different types of problems affecting the environment and
recent advances in environmental protection strategies. The role and potential of
rhizospheric microorganisms in plant growth in disturbed soils is presented. Agro-
industrial wastes and municipal solid wastes management options are discussed;
the various sustainable solutions are also highlighted. The health effects of dyes
and their different remedial treatment process and also the potential of peroxidases
for treatment of dyes are discussed. The resistance and transfer genes in microor-
ganisms and their molecular detection methods are explained along with the abil-
ity of environmental bacteria to form biofilms. The biochemical attributes for the
assessment of soil ecosystem sustainability and the various methods involved in
genotoxicity testing of environmental pollutants are summarized. Pesticides bio-
remediation strategies from soil and wastewater and the biodegradation of cyano-
bacteria and their toxins are outlined. The cause of Alfalfa damping off and the
characterization of the causal agent are discussed. The significance of biochemical
compounds derived from legumes and rhizobacteria (rhizodeposits) with potential
in biotechnology are explained. The pulp and paper industry is a big sector and gen-
erates large amounts of wastewater; the treatment processes are briefly presented.
The contamination of shooting range soils with heavy metals is a matter of con-
cern and the remediation processes are discussed. The chapter ends with the role of
biopesticides in sustainable agriculture. Thus, whole work concluded the ill effects
of different types of pollution and the waste generated by human activities in the
environment. Current trends involved in the remediation and the technologies used
for this purpose are presented in detail.
Keywords Agriculture • Bioremediation • Environmental protection • Pollutants •
Sustainability
A. Malik, E. Grohmann (eds.), Environmental Protection Strategies for Sustainable
Development, Strategies for Sustainability,
DOI 10.1007/978-94-007-1591-2_1, © Springer Science+Business Media B.V. 2012
Chapter 1
Environmental Protection Strategies:
An Overview
Abdul Malik, Mashihur Rahman, Mohd Ikram Ansari, Farhana Masood
and Elisabeth Grohmann
A. Malik ()
Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim
University, Aligarh-202002, India
e-mail: ab_malik30@yahoo.com
2
1.1 Introduction
Sustainable development is now of primary importance as the key to future use
and management of finite world resources. It recognizes the need for development
opportunities while maintaining a balance between these and the environment. As
stated by the UN Bruntland Commission in 1987, sustainable development should
“meet the needs of the present generation without compromising the ability of fu-
ture generations to meet their own needs, encompasses, e.g. keeping population
densities below the carrying capacity of a region, facilitating the renewal of renew-
able resources, conserving and establishing priorities for the use of non-renewable
resources, and keeping environmental impact below the level required to allow af-
fected systems to recover and continue to evolve”. Environmental sustainability can
be viewed as balancing the “three pillars” of economic and social development with
environmental protection.
Soil and water are basic resources essential for sustainable agriculture. But un-
fortunately very few people realize the importance of conserving and judiciously
utilizing the soil as a basic resource by successfully managing the soil internal, ex-
ternal, renewable or non renewable inputs. Sustainable agriculture is defined as the
successful management of resources for agriculture production to satisfy changing
human needs while maintaining or enhancing the quality of the environment and
conserving natural resources (CGIAR 1989). The key words found in the above
definition are (a) resource management (b) changing human needs (c) environmen-
tal quality and natural resources. While keyword (b) is involved in the concept of
agricultural productivity, keyword (c) is involved in the protection of the environ-
ment concept. Both concepts are combined in keyword (a) which refers to resource
management. Therefore, the basic concept of sustainable is to define the combined
concept of productivity (or profitability) and environmental soundness in farm man-
agement or in agriculture at the national level.
The World Commission on Environment and Development (1987) defined sus-
tainability as “a system which can be considered sustainable if it ensures that today’s
economic development is not at the expense of tomorrow’s development prospects”.
Sustainability is not a fixed phenomenon; it changes with time, requirement,
and places and so on. A system, which is sustainable today, may not be sustainable
tomorrow and vice versa. The Rio declaration of 1992, while enunciating many
principles for sustainability of environment and development, emphasized the fol-
lowing principles most relevant to sustainable agriculture.
• Human beings are at the centre of concern for sustainable development and en-
vironmental issues are best handled with the participative co-operation of all
concerned.
• No development can be considered complete unless it meets the criteria of pro-
ductivity, equity and environmental safety for the present and future generations.
• To ensure development and sustainability, it is necessary to remove all the nega-
tive factors leading to unsustainability.
A. Malik et al.
3
• Environmental policies must form an integral part of development policies and
strategies.
The definition of “sustainable land use” is in consonance with “safe environment”
or “environmental protection”, because harmonization of all the land uses in a given
area guarantees safe environment conditions in the sense of a safe protected habitat
for “human and other living organism”. Therefore sustainable land use creates and
guarantees a safe environment by definition (Blum 1994).
Sustainable Agriculture has several positive points such as lesser use of pesti-
cides and chemical fertilizers because when these are used in excess they not only
pollute the environment but also cause health hazards as these pollutants might be
reaching to the food grains, vegetables, fruits, fodder and milk. However, sustain-
able agriculture should be not equated with subsistence farming and perpetual low
yields.
An alternative to increase agricultural productivity in a sustainable manner, there
is increasing reliance on manipulation of microorganisms that benefit soil and plant
health (Kloepper et al. 1989). Plant growth promoting rhizobacteria (PGPR) com-
prise a diverse group of rhizosphere colonizing bacteria and diazotrophic micro-
organisms which, when grown in association with a plant, stimulate growth of the
host. PGPR can affect plant growth and development indirectly or directly (Glick
1995; Vessey 2003; Banchio et al. 2008). In indirect promotion, the bacteria de-
crease or eliminate certain deleterious effects of a pathogenic organism through var-
ious mechanisms, including induction of host resistance to the pathogen (Van Loon
and Glick 2004; Van Loon 2007). In direct promotion, the bacteria may provide
the host plant with synthesized compounds; facilitate uptake of nutrients; fix at-
mospheric nitrogen; solubilize minerals such as phosphorus; produce siderophores,
which solubilize and sequester iron; synthesize phytohormones, including auxins,
cytokinins, and gibberellins, which enhance various stages of plant growth; or syn-
thesize enzymes that modulate plant growth and development (Lucy et al. 2004;
Gray and Smith 2005).The large-scale application of PGPR to crop as inoculants
would be attractive as it would substantially reduce the use of chemical fertilizers
and pesticides, which often pollute the environment. This has a heavy impact on the
natural and human environment, as well as on human health, through the pollution
of soils, waters, and the whole food supply chain.
The use of arbuscular mycorrhizal fungi in ecological restoration projects has
been shown to enable host plant establishment on degraded soil and improve soil
quality and health (Jeffries et al. 2003; Siddiqui et al. 2008). Disturbance of native
plant communities in desertification-threatened areas is often followed by degrada-
tion of physical and biological soil properties, soil structure, nutrient availability
and organic matter. When restoring disturbed land it is essential to not only replace
the above ground vegetation but also the biological and physical soil properties
(Jeffries et al. 2003). A relatively new approach to restore land and protect against
desertification is to inoculate the soil with arbuscular mycorrhizal fungi (AMF)
with the reintroduction of vegetation. The benefits observed were an increased plant
growth and soil nitrogen content, higher soil organic matter content and soil aggre-
1 Environmental Protection Strategies: An Overview
4
gation. The improvements were attributed to the higher legume nodulation in the
presence of AMF, better water infiltration and soil aeration due to soil aggregation.
Inoculation with native AM fungi increased plant uptake of phosphorus, improving
plant growth and health. AM fungi can contribute to plant growth, particularly in
disturbed or heavy metal contaminated sites, by increasing plant access to rela-
tively immobile minerals such as P (Vivas et al. 2003; Yao et al. 2003; Gosling et
al. 2006), improving soil texture by binding soil particles into stable aggregates
that resist wind and water erosion (Rillig and Steinberg 2002; Steinberg and Rillig
2003; Siddiqui et al. 2008) and by binding heavy metals into roots that restricts their
translocation into shoot tissues (Dehn and Schuepp 1989; Kaldorf et al. 1999; Gos-
ling et al. 2006). Furthermore, the fungi can accelerate the revegetation of severely
degraded lands such as coal mines or waste sites containing high levels of heavy
metals (Marx 1975; Marx and Altman 1979; Gaur and Adholeya 2004).
For successful biotechnological application in the field, the selection of micro-
bial inoculums is very important as maximum on-site benefits will only be obtained
from inoculation with efficient fungi and/or bacteria in compatible host/microor-
ganism/site combinations. The basic criteria for subsequent selection and later ap-
plication of microbial inoculum useful for plant-growth promotion is cultivability
and fast multiplication of microorganisms. Information on critical factors influenc-
ing plant-microbe-pollutant interactions in soils could lead to an improved selection
of microbial inoculum for a microbial-assisted bioremediation. Thus an improved
fundamental knowledge of physiological traits of rhizosphere microorganisms and
their impact on rhizosphere processes, which are especially relevant for the reme-
diation of disturbed soils will be essential to allow an increased and successful use
of microbial inoculum in the field.
Agro-industrial wastes can be generally organized into different categories, such
as food processing wastes, energy crops and biofuel production wastes and crop
residues. Agricultural wastes comprise almost 15% of total waste generated by each
country (Hsing et al. 2001; Arvanitoyannis et al. 2006). It has been estimated that
approximately 30% of global agricultural products are becoming residues and re-
fuses. Large volumes of solid and liquid wastes are generated from the agro-pro-
cessing industries. Waste from agriculture and food processing can become one of
the most serious sources of pollution (Di Blasi et al. 1997; Monspart-Sényi 2007).
The efficient utilization of agro-waste can lead to improvement in agricultural yield
and environmental health by decreasing the pollution caused by the agro wastes.
The most commonly used methods by which the agro wastes are managed in differ-
ent countries are: landfilling, incineration, composting and recycling.
Agro-industrial wastes and by-products (substances that originate during pro-
cessing) can be further utilized in several other ways. Many by-products of the
agro industry can be fed to animals directly as such without any modification or
can be used after fermentation of the agro-residues. Recovery of by-products for
use as animal feed can help agro industry save money by reducing waste discharges
and can cut waste management costs and also can prevent environmental pollu-
tion. Microorganisms are grown on food processing by-products and utilized in the
production of enzymes, single cell protein, amino acids, lipids, carbohydrates and
A. Malik et al.
5
organic acids. Agro-by-products can also be beneficially used as soil conditioner or
fertilizer. Over the last century, energy consumption has increased beyond control
as a result of growing world population and industrialization (Sun and Cheng 2002).
The limited number of known fossil fuel deposits and the threat to environment due
to the use of these fossil fuels has made it essential to look for alternative and re-
newable sources of fuels. Renewable energy sources, such as ethanol, methane, bio-
hydrogen can be produced by fermentation of sugars. Owing to diminishing natural
oil and gas resources, interest in the bioconversion of renewable cellulosic biomass
into fuel ethanol as an alternate to petroleum is rising around the world (Stevenson
and Weimer 2002; Reddy et al. 2010). Biomass is the earth’s most attractive alterna-
tive among fuel sources and sustainable energy resource. Agro-industrial residues
produce ethanol, bioethanol, a product of high potential value containing minor
quantities of soluble sugars, pectin, proteins, minerals and vitamins. Bioethanol
produced from renewable biomass has received considerable attention in current
years. Using ethanol as a gasoline fuel helps to alleviate global warming and en-
vironmental pollution. They also have potential to produce biogas under anaerobic
fermentation conditions. Biological conversion offers a potential for radical techni-
cal advances through application of the powerful tools of modern biotechnology to
realize truly low costs. In the last few decades, vermicomposting technology has
been arising as a sustainable tool for the efficient utilization of the agro-industrial
processing wastes and to convert them into value added products for land restora-
tion practices. The product of the process, i.e., vermicompost is humus like, finely
granulated and friable material which can be used as a fertilizer to reintegrate the
organic matter to the agricultural soils (Garg and Gupta 2009).
Adsorption process has been proven one of the best water treatment technolo-
gies around the world and activated carbon is undoubtedly considered as universal
adsorbent for the removal of diverse types of pollutants from wastewater. However,
widespread use of commercial activated carbon is sometimes restricted due to its
higher costs. Attempts have been made to develop inexpensive adsorbents utilizing
numerous agro-industrial and municipal waste materials. Use of waste materials as
low-cost adsorbents is attractive due to their contribution in the reduction of costs
for waste disposal, therefore contributing to environmental protection. Agricultural
materials have cellulose, hemicelluloses, lignin, sugars, proteins, and starch con-
taining various functional groups that facilitate metal complexion which in turn
helps in the sequestration of heavy metals (Hashem et al. 2007; Bhatnagar and Sil-
lanpää 2010).
By-products of plant food processing may be used because of their favourable
nutraceutical properties. There are large varieties of value-added compounds in the
by-products and wastes of biological origin. These products may be used as such or
may serve as a starting material for the preparation of novel compounds like anti-
oxidants, carbohydrates, dietary fibers, fat and oils, pigments, proteins and starch.
Ensuring environmental safety and sustainable development through waste utiliza-
tion aims to ensure that the development needs of the present do not compromise
the needs of future generations (Ahmad et al. 2010).
1 Environmental Protection Strategies: An Overview
6
A variety of synthetic dyestuffs released by the textile industry pose a threat to
environmental safety. Azo dyes account for the majority of all dyestuffs produced,
because they are extensively used in the textile, paper, food, leather, cosmetics and
pharmaceutical industries. Dye-house effluent typically contains only 0.6–0.8 g L
–1
dye, but the pollution it causes is mainly due to durability of the dyes in the waste-
water (Jadhav et al. 2007). Existing effluent treatment procedures are unable to
remove recalcitrant azo dyes completely from effluents because of their color fast-
ness, stability and resistance to degradation. Therefore, it is necessary to search for
and develop effective treatments and technologies for the decolorization of dyes in
such effluents. Various physical/chemical methods, such as adsorption, chemical
precipitation, photolysis, chemical oxidation and reduction, electrochemical treat-
ment, have been used for the removal of dyes from wastewater. Moreover, there
are many reports on the use of physicochemical methods for the color removal
from dye containing effluents (Vandevivere et al. 1998; dos Santos et al. 2007;
Wang et al. 2009a, b, c). Several physicochemical methods have been used for the
removal of dyes from wastewater effluent. However, implementation of physical/
chemical methods has the inherent drawbacks of being economically unfeasible
(as they require more energy and chemicals), being unable to completely remove
the recalcitrant azo dyes and/or their organic metabolites, generating a significant
amount of sludge that may cause secondary pollution problems, and involving com-
plicated procedures (Forgacs et al. 2004; Zhang et al. 2004). However, microbial
or enzymatic decolorization and degradation is an eco-friendly cost-competitive
alternative to chemical decomposition process that could help reduce water con-
sumption compared to physicochemical treatment methods (Verma and Madamwar
2003; Rai et al. 2005).
The use of microorganisms for the removal of synthetic dyes from industrial
effluents offers considerable advantages. The process is relatively inexpensive,
the running costs are low and the end products of complete mineralization are not
toxic. The various aspects of the microbiological decomposition of synthetic dyes
have been previously reviewed by Stolz (2001). Besides the traditional wastewater
cleaning technologies, other methods have been employed in the microbial decol-
orization of dyes. For instance, an activated sludge process was developed for the
removal of Methyl violet and Rhodamine B from dyestuff effluents, using micro-
organisms that were derived from cattle dung (Kanekar and Sarnaik 1991). Also in
biofilms, efficient biodegradation of Acid Orange 7 has been demonstrated (Harmer
and Bishop 1992; Zhang et al. 1995). Azo dyes did not inhibit the capacity of bio-
films in the removal of organics from wastewater (Fu et al. 1994). A multistage
rotating biological contactor was used for the biodegradation of azo dyes, where an
azo dye assimilating bacterium was immobilized in the system (Ogawa and Yatome
1990).
The emergence of bacterial antibiotic resistances as a consequence of the wide-
scale use of antibiotics by humans has resulted in a rapid evolution of bacterial
genomes. Mobile genetic elements such as transferable plasmids, transposons and
integrons have played a key role in the dissemination of antibiotic resistance genes
amongst bacterial populations and have contributed to the acquisition and assembly
A. Malik et al.
7
of multiple antibiotic resistance in bacterial pathogens (Tschäpe 1994; Salyers and
Shoemaker 1994; Mazel 2006; Rahube and Yost 2010). Bacteria resistant to mul-
tiple antibiotics are not restricted to clinical environments but can easily be isolated
from different environmental samples and food (Perreten et al. 1997; Feuerpfeil
et al. 1999; Dröge et al. 2000). There is substantial movement of antibiotic resis-
tance genes and antibiotic resistant bacteria between different environments. In
assessing the antibiotic resistance problem, a number of factors can be identified
which have contributed to the antibiotic resistance problem: the antibiotic itself and
the antibiotic resistance trait (Levy 1997; Andersson and Hughes 2010). The genetic
plasticity of bacteria has largely contributed to the efficiency by which antibiotic
resistance has emerged. However, horizontal gene transfer events have no a priori
consequence unless there is antibiotic selective pressure (Levy 1997). Since bac-
teria circulate between different environments and different geographic areas, the
global nature of the problem of bacterial antibiotic resistances requires that data on
their prevalence, selection and spread are obtained in a more comprehensive way
than before (Shaw et al. 1993; Wright 2010). DNA probes and PCR-based detec-
tion systems allow us not only to analyze the dissemination of antibiotic resistance
genes in the culturable fraction of bacteria but also to extend our knowledge to the
majority of bacteria which are not accessible to traditional cultivation techniques
(Smalla and van Elsas 1995; Heuer et al. 2002). Studies on the dissemination of the
most widely used marker gene, nptII, in bacteria from sewage, manure, river wa-
ter and soils demonstrated that in a high proportion of kanamycin-resistant enteric
bacteria the resistance is encoded by the nptII-gene (Leff et al. 1993; Smalla et al.
1993; Lynch et al. 2004).
In recent years, approaches have been implemented to characterize the diversity
and prevalence of resistance in soil bacteria-the soil antibiotic resistome-as an im-
portant reservoir of resistance (Wright 2007). Riesenfeld et al. (2004) investigated
resistance in the soil, concentrating on unculturable organisms, bacteria that have
yet to be characterized and thus underappreciated because of challenging culture
conditions (Riesenfeld et al. 2004). By creating a functional metagenomic library
(Handelsman 2004) in which cloned genomic fragments were expressed from DNA
isolated directly from soil and selecting for resistance, traditional challenges as-
sociated with studying genes of unknown sequence were circumvented. Specifi-
cally, these functional analyses revealed novel antibiotic resistance proteins that
were previously of unknown function and unrecognizable by sequence alone. Thus,
this work not only allowed for the identification of aminoglycoside N-acetyltrans-
ferases, the O-phosphotransferases, and a putative tetracycline efflux pump but also
a construct with a novel resistance determinant to the aminoglycoside butirosin (Ri-
esenfeld et al. 2004). This shows the power of the functional metagenomic approach
when applied to a search of activity with a highly selectable phenotype such as
antibiotic resistance.
It is important to remark that several antibiotics are produced by environmental
microorganisms (Waksman and Woodruff 1940). Conversely, antibiotic resistance
genes, acquired by pathogenic bacteria through Horizontal Gene Transfer (HGT)
have been originated as well in environmental bacteria (Davies 1997), although
1 Environmental Protection Strategies: An Overview
8
they can evolve later on under strong antibiotic selective pressure during the treat-
ment of infections (Martinez and Baquero 2000; Martinez et al. 2007). To under-
stand in full the development of resistance, we will thus need to address the study of
antibiotics and their resistance genes, not just in clinics but in natural non-clinical
environments also (Martinez 2008).
Understanding heavy metal resistance in natural ecosystems may help as well
to understand antibiotic resistance in the environment. The elements involved in
the resistance to heavy metals are encoded in the chromosomes of bacteria like
Ralstonia metallidurans (Mergeay et al. 2003), which are well adapted for surviv-
ing in naturally heavy metals-rich habitats (e.g. volcanic soils). However, strong
selective pressure due to anthropogenic pollution has made that these chromosom-
ally-encoded determinants are now present in gene-transfer units, so that they can
efficiently spread among bacterial populations (Silver and Phung 1996, 2005; Nies
2003). Similarly, antibiotic resistance genes that were naturally present in the chro-
mosomes of environmental bacteria (D’Acosta et al. 2006; Wright 2007; Fajardo
and Martinez 2008) are now present in plasmids that can be transferred to human
pathogens. It has been highlighted that the contact of bacteria from human-associ-
ated microbiota with environmental microorganisms in sewage plants or in natural
ecosystems is an important feature to understand the emergence of novel mecha-
nisms of resistance in human pathogens (Baquero et al. 2008). A key issue for this
emergence will be the integration of antibiotic resistance genes in gene-transfer
elements (e.g. plasmids), a feature that is favoured by the release of antibiotics in
natural ecosystems (Cattoir et al. 2008).
It was suggested by Rysz and Alvarez (2004) that ARGs themselves could be
considered as environmental “pollutants”, since they are widely distributed in
various environmental compartments, including wastewater and sewage treatment
plants (STPs), surface water, lagoon water of animal production areas, aquaculture
water, sediments and soil, groundwater, and drinking water. Pruden et al. (2006)
have also pointed out that ARGs may be thought as emerging “contaminants”, for
the public health problems resulting from the widespread dissemination of ARGs.
So far, the methods used for detection, typing, and characterization of ARGs have
covered, but not been limited to, specific and multiplex polymerase chain reaction
(PCR), real-time PCR, DNA sequencing, and hybridization-based techniques in-
cluding microarray.
The widespread pollution of soils is an increasingly urgent problem because of
its contribution to environmental deterioration on a global basis (Bezdicek et al.
1996; Dick 1997; Lal 1997; van Beelen and Fleuren-Kemilá 1997). Until relatively
recently, soil was widely regarded as just an environmental filter ensuring the qual-
ity of both water and atmosphere. However, in the context of the pursuit of sustain-
ability, it is now recognised that soil is not only an effective de-contaminant of
potential pollutants but that its chemical, physical and biological quality must be
maintained (Hornick 1992; Parr et al. 1992). From the point of view of sustainabil-
ity, a high-quality soil is a soil that is capable of producing healthy and abundant
crops; decontaminating the water passing through it; not emitting gases in quantities
detrimental to the environment; and behaving as a mature, sustainable ecosystem
A. Malik et al.
9
capable of degrading organic input (Doran and Parkin 1994; Gregorich et al. 1994;
Brookes 1995; Pankhurst et al. 1995). This view clearly implies that diagnosis of
soil pollution should be carried out on the basis of observed alterations in the soil
properties controlling the behaviours described above, ideally in a way that allows
any loss of soil quality to be quantified as well as identified qualitatively (Larson
and Pierce 1991; Doran and Parkin 1994).
The concept of soil quality gives rise to more controversy than that of water or
air quality. However, despite the difficulty in providing a definition, the mainte-
nance of soil quality is critical for ensuring the sustainability of the environment
and the biosphere. Literature exhibits a great number of soil quality indices for both
agro-ecosystems and natural or contaminated soils. The book has reviewed some
of the soil quality indices established up to date as well as of the parameters that
make up them, and to offer a reflection on the lack of consensus concerning the
use of these indices. We have focused on those indices including biological param-
eters. The most straightforward index used in the literature is the metabolic quotient
(qCO
2
) (respiration to microbial biomass ratio), widely used to evaluate ecosystem
development, disturbance or system maturity. However, qCO
2
and other indices
integrating only two parameters provide insufficient information about soil quality
or degradation. For this, lately there has been a wide development of multipara-
metric indices that clearly establish differences between management systems, soil
contamination or density and type of vegetation. These indices integrate different
parameters, among which the most important are the biological and chemical ones,
such as pH, organic matter, microbial biomass C, respiration or enzyme activities.
The major part of multiparametric indices has been established based on either,
expert opinion (subjective), or using mathematical statistics methods (objective).
Molecular indicators have not yet been used for soil quality indices establish-
ment. However, the development of genomic, transcriptomic or proteomic meth-
odologies could have importance in the evaluation of soil quality, not only in a
diversity sense but also in a functional way. These methods can provide information
about what is the role of specific microorganisms and their enzymes in key pro-
cesses related to soil functionality. Despite of the great diversity of indices, they
have never been used on larger scales, nor even in similar climatological or agro-
nomic conditions. The lack of applicability of soil quality indices resides on: (i)
poor standardization of some methodologies; (ii) some methods are out of reach in
some parts of the world; (iii) spatial scale problems (soil heterogeneity); (iv) poor
definition of soil natural conditions (climate and vegetation); and (v) poor definition
of soil function to be tested for soil quality.
New chemicals are being added each year to the existing burden of toxic sub-
stances in the environment. This has led to increased pollution of ecosystems as
well as deterioration of the air, water and soil quality. Excessive agricultural and in-
dustrial activities adversely affect biodiversity, threatening the survival of species in
a particular habitat as well as posing disease risks to humans. Test systems that help
in hazard prediction and risk assessment are important to assess the genotoxic po-
tential of chemicals before their release into the environment or for commercial use.
1 Environmental Protection Strategies: An Overview
10
Currently, standardized prokaryotic genotoxicity procedures include the Ames
test, the umu-test and the SOS chromotest which are based on genetically engi-
neered Salmonella typhimurium strains. Tests with eukaryotic cells or organisms
might be more relevant for human and ecological risk assessment, but generally
they are much more time-consuming. Several tests have been developed using the
integrity of DNA as an unspecific endpoint of genotoxicity e.g. Comet Assay, Al-
kaline DNA-eluation assay, DNA alkaline unwinding assay, UDS-assay; the Comet
assay probably the most cost-efficient test among these.
To date, numerous in vivo tests have been developed that take into account uptake
and elimination, internal transport and metabolism of pollutants. In vivo methodolo-
gies for assessing genotoxicity as part of routine toxicity testing are now available,
and it is time to move to a regulatory paradigm that includes an assessment of in vivo
factors that determines genotoxic outcome, where possible integrating genotoxic-
ity determinations into routine toxicity tests. In vivo mutation assays are based on
the following principles: (1) selection of mutants based on enzymatic activity of an
endogenous enzyme in cells isolated after exposure in vivo and then cultured in vitro
(e.g., the hprt or tk assays in lymphocytes), (2) recovery of a reporter transgene that
is subsequently tested for expression in a recipient cell in vitro (e.g., the lacI or lacZ
transgenic rodent systems), or (3) antibody-based methods to identify structural
alterations in cell or cell-surface proteins (e.g., the glycophorin A or T-cell receptor
assays) (MacGregor 1994). Newer technologies such as transcriptomics, proteomics
and metabolomics provide the opportunity to gain insight into genotoxic mecha-
nisms and also to provide new markers in vitro and in vivo. There is also an increas-
ing number of animal models with relevance to genotoxicity testing. These types of
models will undoubtedly have an impact on genotoxicity testing in the future.
Cyanobacteria are a widespread group of organisms colonizing all ecosystems.
They are common inhabitants of freshwater bodies throughout the world and sev-
eral of them form surface scums (blooms). Under favourable conditions several
species of cyanobacteria may become dominant in the phytoplankton of water bod-
ies. Cell densities may reach many millions per litre (Chorus and Bartram 1999).
Cyanobacteria are known to produce several metabolites significant from the public
health perspective of acute exposure: lipopolysaccharides (Stewart et al. 2006), and
cytotoxic, tumor promoting and enzyme inhibiting metabolites like cyclic depsip-
eptides, cyclic peptides (anabaenopeptinsandnostophycins), linear peptides (aerugi-
nosins and microginins) (Bickel et al. 2001; Forchert et al. 2001; Welker and Von
Dohren 2006). A neurotoxin non-protein amino acid ( N-methylamino-l-alanine,
BMAA), widely produced among cyanobacteria (Cox et al. 2005), has been as-
sociated with neurodegenerative disease such as Alzheimer’s disease, amyotrophic
lateral sclerosis/parkinsonism-dementia complex (Murch et al. 2004). Many cyano-
bacterial species can produce several categories of powerful toxins that are unique
to this group of organisms, with the exception of saxitoxins. Cyanotoxin poisoning
in humans was mainly caused by three toxic groups: microcystins (MCYSTs), cyl-
indrospermopsin and anatoxin-a(ANA-a), and occurred through exposure to con-
taminated drinking water supplies (Annadotter et al. 2001; Falconer 2005), recre-
A. Malik et al.
11
ational waters (Chorus and Bartram 1999; Behm 2003), medical dialysis (Azevedo
et al. 2002).
Cyanobacteria toxins have quickly risen in infamy as important water contami-
nants that threaten human health (Svrcek and Smith 2004). Toxins are introduced
to the environment, in general, through the rupture of algal cells which may arise
by the effect of certain substance used during water treatment (Carmichael 1992)
and/or in different water management processes, e.g. in algaecide treatment of the
natural medium, pumping of raw water, conveyance of raw water.
Microcystins are chemically stable in water and cannot be effectively removed
by conventional water treatment processes. It is possible that the treatment process
may cause cell lysis and release the intracellular metabolites containing toxins (Tsu-
ji et al. 1997; Chow et al. 1999). The water treatment process for removing algae
and microcystins from source water urgently needs to be studied.
Microcystins are normally present inside cyanobacterial cells and enter the
surrounding water after cell lysis. The major route of detoxification of MC-LR is
probably biodegradation (Lahti et al. 1997; Miller and Fallowfield 2001; Ishii et al.
2004). Interestingly, bacteria like Pseudomonas spp. isolated from the surface water
of lakes, rivers and dams decreased microcystins. Takenaka and Watanabe (1997)
isolated four kinds of bacteria classed in the genera of Pseudomonas, Citrobacter,
Enterobacter and Klebsiella from the surface water of a Japanese lake where a
heavy water bloom occurs every year, and tested the bacterial degradation ability
of microcystin LR. They found that only a bacterium identified as Pseudomonas
aeruginosa degraded microcystin LR. In some laboratory studies, dissolved micro-
cystins have been rather resistant to degradation.
Biodegradation after a lag period of a few days or weeks has been the most im-
portant means of detoxification in laboratory experiments. Microcystins produced
in natural waters can be degraded by indigenous microorganisms, although the pro-
cess occasionally is slow and may require a period of adaption (Cousins et al. 1996).
A new biological treatment method of purification of source water quality in a
eutrophic lake using indigenous enrichment microbes by artificial media was used
by Ji et al. (2009). The test of algae and microcystins degradation by enrichment mi-
crobes on the artificial media (assembled medium, elastic medium and non-woven
fabric medium) revealed that the average removal efficiency of chlorophyll-a was
above 60%, and the removal effect of microcystins was 40–67%. Enrichment mi-
crobes on the artificial media could effectively degrade algae and microcystins in
Lake Taihu. PCR based results showed that there were algae-lysing bacteria in the
natural source water (Ji et al. 2009).
Pesticides are extensively used to increase agricultural production by prevent-
ing losses due to pests. However, some are among the highly persistent, toxic, and
bioaccumulative contaminants in the environment generally referred to as persistent
organic pollutants (POPs) (WHO 2009). These toxicants get into the human body
through the food chain, and can cause serious health problems (Albert and Rendon
1988).
Some pesticides are known to resist biodegradation and therefore, they can be re-
cycled through food chains and produce a significant bioaccumulation at the higher
1 Environmental Protection Strategies: An Overview
12
end of the chain (Shukla et al. 2006). For this reason, pesticide residue analysis in
environmental samples has received increasing attention in the last few decades,
resulting in numerous environmental monitoring programs in various countries for
a broad range of pesticides. A common consequence of such persistent pollution is
the contamination of surface waters with pesticide residues. This calls for urgent at-
tention in two areas: (a) re-evaluation of environmental persistence and risks of cur-
rently registered and applied pesticides, and (b) thorough monitoring of potentially
water-contaminating pesticides in surface waters and in natural bodies (Mukherjeez
and Gopal 2002; Donald et al. 2007; Maloschik et al. 2007).
Biodegradation is a natural process, where the degradation of a xenobiotic
chemical or pesticide by an organism is primarily a strategy for their own survival.
Most of these microbes work in natural environment but some modifications can be
brought about to encourage the organisms to degrade the pesticide at a faster rate in
a limited period. This capability of microbes is sometimes utilized as technology for
removal of contaminant from actual site. Knowledge of physiology, biochemistry
and genetics of the desired microbe may further enhance the microbial process to
achieve bioremediation with precision and with limited or no scope for uncertainty
and variability in microbe functioning. Genes encoding enzymes for degradation
of several pesticides, have been identified, which will provide new inputs in under-
standing the microbial capability to degrade a pesticide and develop a super strain
to achieve the desired result of bioremediation in a short time (Singh 2008). Geneti-
cally modified microbes are used to enhance the capability of degradation. Yet, the
use of genetic engineering for the use in environment is still controversial because
an adverse genotype can be readily mobilized in the environment. In a development
of technology for degradation following points should be taken care of i.e. (1) het-
erogeneity of contaminant, (2) concentration of contaminant and its effect on bio-
degradative microbe, (3) persistence and toxicity of contaminant, (4) behaviour of
contaminant in soil environment and (5) conditions favourable for biodegradative
microbe or microbial population. The use of technology at the actual site requires
(1) the knowledge of the natural bioprocess at the contaminated site, (2) detailed
and valid data of microbial biodegradation developed in the laboratory, (3) monitor-
ing of the onsite biodegradation process.
Most of the bioremediation technologies for the field are designed to remove the
pollutant once it is generated or released into the environment. Usually, these tech-
nologies include, bioaugmentation (addition of organism or enzyme to the contami-
nant), biostimulation (use of nutrients to stimulate naturally occurring organisms),
biofilters (removal of organic gases by passing air through compost or soil contain-
ing microorganism), bioreactors (treatment of contaminant in a large tank contain-
ing organism or enzyme), bioventing (involves the venting of oxygen through soil to
stimulate the growth of natural microorganisms capable of degrading contaminant),
composting (involves mixing of contaminant with compost containing bioremedia-
tion organisms) and landfarming (use of farming, tilling and soil amendment tech-
niques to encourage the growth of bioremediation organism at contaminated site).
Forage legumes are essential for efficient animal-based agriculture worldwide.
Besides providing high quality feed for livestock, they are a key component in the
A. Malik et al.
13
sustainability of crop-pasture rotations. Their value lies essentially in their ability
to fix nitrogen (N
2
) in symbiosis with root nodule soil bacteria, collectively called
rhizobia. Microbial-based strategies that improve forage legume establishment and
optimize N
2
fixation have been deployed worldwide through rhizobial inoculant
technology (Catroux et al. 2001; Höfte and Alteir 2010). However, the study of
rhizospheric bacteria for plant-growth promotion remains a challenge (Handelsman
et al. 1990; Jones and Samac 1996; Xiao et al. 2002; Villacieros et al. 2003; Höfte
and Alteir 2010).
Alfalfa, Medicago sativa L., is among the most prized of forages and is grown
worldwide as a feed for all classes of livestock. It is one of man’s oldest crops,
and its cultivation probably predates recorded history. In addition to its versatility
as a feed, alfalfa is well known for its ability to improve soil structure and, as a
legume, is an effective source of biological nitrogen. Their symbiotic association
with rhizobia makes the atmospheric nitrogen available for themselves and other
crops in the rotation. Alfalfa ( Medicago sativa) is an important forage crop owing
to its unique characteristics: high yield of excellent quality forage, hydric-stress
tolerance and good persistence. However, rapid seedling emergence and adequate
pasture establishment are crucial to maximize its potential. Like most crops, alfalfa
is attacked by many disease-causing organisms. Seedlings as well as seeds, stems,
leaves, and roots of older plants all serve as food sources for a number of disease-
causing organisms.
Seedling diseases caused by soil-borne pathogens, primarily Macrophomina
phaseolina (Tassi) Goid and other Oomycetes, are a critical factor, which limits
alfalfa establishment causing pre- and/or post-emergence seedling damping-off.
Damping off is a name given to a condition where seeds are killed before germina-
tion (pre emergence) or seedlings (post emergence) are stunted or collapse and die.
Seeds destroyed before germination are discolored and soft. After seed germination,
symptoms include brown necrotic lesions along any point of the seedling. Lesions
that girdle the young root or stem lead to plant death. Partially girdled plants, as
well as those subject to continued root tip necrosis, may be stunted and yellowish
in color to varying degrees.
The indiscriminate use of chemical fungicides is not recommended for the man-
agement of alfalfa diseases because of their collateral adverse effects on the envi-
ronment, along with negative effects on animal and human health. Moreover, their
efficacy has been reduced by the appearance of microbial resistance (Sanders 1984;
Cook and Zhang 1985; Quagliotto et al. 2009) and their detrimental effect on the
biological nitrogen fixation by rhizobia. A high-density sowing practice is usually
the strategy followed by farmers to cope with alfalfa damping-off, but this approach
significantly increases pasture establishment costs. In recent years, plant growth
promoting rhizobacteria (PGPR) have been extensively examined for their role in
biomanagement of pathogens. Although there are other bacterial species that have
been recognized as biocontrol agents against soil-borne pathogens of agricultural
crops (Glick et al. 1999), fluorescent pseudomonads make up a dominant popula-
tion in the rhizosphere and possess several properties that have made them potential
plant growth promoting biocontrol agents of choice. They have been reported for
1 Environmental Protection Strategies: An Overview
14
biological control of different fungal species such as Rhizoctonia, Fusarium, Scle-
rotium, Pythium and Macrophomina (Negi et al. 2005; Höfte and Alteir 2010). The
broad spectrum of the antagonistic activity of pseudomonads is executed by the
secretion of a number of metabolites including antibiotics (Haas and Keel 2003),
volatile hydrogen cyanide (HCN) (Bhatia et al. 2003), siderophores (Gupta et al.
2002), lytic enzyme chitinases and β-1,3 glucanases (Lim and Kim 1995). There-
fore, these attributes make fluorescent pseudomonads effective biocontrol agents.
In addition, the use of Bacillus spp. (Handelsman et al. 1990) and Streptomyces spp.
(Jones and Samac 1996; Xiao et al. 2002; Bakker et al. 2010) has been explored to
control alfalfa seedling damping-off.
Bacterial biofilms are complex communities of microorganisms embedded in a
self-produced matrix and adhering to inert or living surfaces (Costerton et al. 1999).
Biofilms have been observed on a variety of surfaces and in a variety of niches,
and are considered to be the prevailing microbial lifestyle in most environments.
From a medical perspective, biofilm associated bacteria on implants or catheters
are of great concern because they can cause serious infections. For the food indus-
try in particular, the formation of biofilms on food and food processing surfaces,
and in potable water distribution systems, constitutes an increased risk for product
contamination with spoilage or pathogenic micro-flora (Carpentier and Cerf 1993;
Donlan 2002). The development of biofilms can be seen as a five-stage process
(Stoodley et al. 2002): (1) initial reversible adsorption of cells to the solid surface,
(2) production of extracellular polymeric matrix substances resulting in an irrevers-
ible attachment, (3) early development of biofilm architecture, (4) maturation, and
(5) dispersion of single cells from the biofilm. The bacterial phenotype in this dif-
ferentiated, complex, mature biofilm differs profoundly from that in the planktonic
population. One difference with major implications is the increased resistance of
biofilm bacteria towards antimicrobial agents (Lewis 2001).
Fouling of nanofiltration (NF) and reverse osmosis (RO) membranes is a wide-
spread problem that severely limits membrane performance in many water treat-
ment applications (Ridgway and Flemming 1996). Biofouling is especially criti-
cal in wastewater reclamation because municipal wastewater effluents can contain
significant concentrations of bacteria, dissolved organics, and nutrients (Ridgway
et al. 1983; Bailey et al. 1974). When oligotrophic conditions prevail and/or re-
sidual disinfectant is maintained, bacterial growth and replication are minimal.
Hence, fouling is almost certainly caused by deposition of cell debris and effluent
organic matter onto the membrane surface, which leads to a continuous increase in
pressure drop across the membrane (Subramani and Hoek 2008).When nutrients
are abundant and/or there is no residual disinfectant bacterial growth and exopoly-
mer production are most likely the dominant causes of fouling. In this case, viable
bacterial cell deposition and nutrient concentration polarization in regions of stag-
nant cross-flow (such as where feed spacers contact the membrane surface) initiate
fouling, but rapid biogrowth and exopolymer production increase module pressure
drop by clogging feed spacer voids and increasing cross-flow drag (Vrouwenvelder
et al. 2009a, b). Now a number of techniques are used to detect biofilm formation.
Most commonly used technique are staining with dyes like crystal violet, fluores-
A. Malik et al.
15
cein isothiocyanate (FITC) or tetramethyl rhodamine isothiocyanate (TRITC) and
Cyanine (CY5) or the GFP labeling of the bacteria used in biofilm formation (Neu
et al. 2001). Recently some other techniques have been used to directly visualize the
biofilm in the environment like laser scanning microscopy (LSM), magnetic reso-
nance imaging (MRI), scanning transmission X-ray microscopy (STXM), Raman
microscopy (RM), surface-enhanced Raman scattering (SERS) and atomic force
microscopy (AFM) (Neu et al. 2010; Ivleva et al. 2010; Wright et al. 2010).
The rhizosphere of a plant is a zone of intense microbial activity. Rhizobacteria
that exert beneficial effects on plant growth and development are referred to as plant
growth promoting rhizobacteria (PGPR) because their application is often associ-
ated with increased rates of plant growth, development and yield. PGPR can affect
plant growth directly or indirectly. Indirect promotion of plant growth occurs when
introduced PGPR lessen or prevent deleterious effects of one or more phytopatho-
genic organisms in the rhizosphere. The direct promotion of plant growth by PGPR
may include the production and release of secondary metabolites such as plant
growth regulators (phytohormones) or facilitating the uptake of certain nutrients
from the root environment. Kloepper et al. (1991) indicated that different strains of
PGPR can increase crop yields, control root pathogens, increase resistance to foliar
pathogens, promote legume nodulation, and enhance seedling emergence. Co-inoc-
ulation of legumes with rhizobia and PGPR is even more effective for improving
nodulation and growth of legumes.
It is well known that the rhizosphere is characterized by biological, chemical
and physical interactions between plants, microorganisms and soil, and that these
interactions have an effect on nutrition, growth and the general health of the plants
and consequently on their productivity. Actually, several authors suggest that the
plants control these interactions with the rhizodeposition or root exudation of or-
ganic compounds. Indeed, plant roots release a wide range of organic compounds
and most of them are involved in the nutrient acquisition mechanisms (Neumann
and Römheld 1999). Three broad types of rhizodeposits can be determined (Brady
and Weil 1999). First, low-molecular-weight organic compounds are passively ex-
uded by root cells, including organic acids, sugars, amino acids, and phenolic com-
pounds. Second, high-molecular-weight mucilages actively secreted by root-cap
cells and epidermal cells near apical zones form a substance called mucigel when
mixed with microbial cells and clay particles. Third, cells from the root cap and
epidermis continually slough off as the root grows or get digested by bacteria. These
lysates enrich the rhizosphere with a wide variety of cell contents. High molecular
weight compounds such as carbohydrates, proteins and enzymes and low molecular
weight compounds such as organic acids, phenols and amino acids are exuded into
the rhizosphere changing the physical, chemical and biological properties of the
rhizosphere and enhance adaptation to particular environments (Jones et al. 2004,
2009).
Most of the legumes are used as forages, however during the last few years they
have been found to contain medicines, nutraceuticals, and pesticides to promote
their use as new bio-functional crops. These legumes are important sources of for-
age, nutritional supplements, medicines, pesticides, primary and secondary metabo-
1 Environmental Protection Strategies: An Overview
16
lites plus other industrial and agricultural raw material (Graham and Vance 2003).
Legumes also provide essential minerals required by humans (Grusak 2002a) and
produce health promoting secondary compounds that can protect against human
cancers (Grusak 2002b; Madar and Stark 2002) and protect the plant against the on-
slaught of pathogens and pests (Dixon et al. 2002; Ndakidemi and Dakora 2003). In
addition to their blood cholesterol-reducing effect (e.g. Andersen et al. 1984), grain
legumes generally also have a hypoglycemic effect, reducing the increase in blood
glucose after a meal and, hence, blood insulin. Legumes are, therefore, included in
the diet of insulin-dependent diabetics (Jenkins et al. 2003). Genomics approaches,
including metabolomics and proteomics, are essential to understanding the meta-
bolic pathways that produce these antinutritional compounds and to eliminating
these factors from the plant. The general theme of improvement of food and feed
represents a clear vision for the future for legume genomics, as well as an emphatic
statement directed primarily toward the public, who will be the ultimate beneficiary
of genomic activities. This unified theme combines several areas of research. First,
it recognizes the importance of grain legumes (also known as pulses) as essential
sources of dietary protein for humans and animals, as well as health-related phyto-
chemicals such as dietary fiber, hormone analogs, and antioxidants. Genomics pro-
vide essential tools to fully understand the molecular and metabolic basis of the syn-
thesis of these compounds, to increase their content in seeds and pods, and to better
manipulate interactions between the plant’s genetic makeup and its environment. A
focus on seeds also underscores the importance of the genomics of reproductive bi-
ology in the development of higher-yielding, more nutritious legume cultivars. With
the advent of biotechnology many plant constituents have found their way in food,
chemical and energy industries. Plants now are used as bioreactors for production
of bioactive peptides, vaccines, antibodies and a range of enzymes mostly for the
pharmaceutical industry. For the chemical industry, plants can be used to produce,
e.g., polyhydroxybutyrate for the production of biodegradable thermoplastics, and
cyclodextrins, which form inclusion complexes with hydrophobic substances (Alt-
man 1999; van Beilen 2008).
The pulp and paper industry involves three basic areas: paper making, paper
converting and printing. The pulp and paper industry converts fibrous raw materi-
als into pulp, paper and paperboard. About 500 different chlorinated organic com-
pounds have been identified including chloroform, chlorate, resin acids, chlorinated
hydrocarbons, phenols, catechols, guaiacols, furans, dioxins, syringols, vanillins,
etc. (Suntio et al. 1988; Freire et al. 2000). In wastewater, these compounds are esti-
mated collectively as adsorbable organic halides (AOX). These compounds are usu-
ally biologically persistent, recalcitrant and highly toxic to the environment (Baig
and Liechti 2001; Thompson et al. 2001). The toxic effects of AOX range from
carcinogenicity, mutagenicity to acute and chronic toxicity (Savant et al. 2006).
Increased awareness of the harmful effects of these pollutants has resulted in
stringent regulations on AOX discharge into the environment (Bajpai and Bajpai
1994; Deshmukh et al. 2009). There are several modifications made to reduce the
generation of chlorinated organic compounds from bleach plant effluents using one
or more of the following strategies: (1) removing more lignin before starting the
A. Malik et al.
17
chlorination, i.e., reducing the kappa number of unbleached pulp, (2) modifying
the conventional bleaching process to elemental chlorine free bleaching (ECF) and
total chlorine free bleaching (TCF). These methods may be physicochemical or
biochemical in nature or a combination thereof (Bajpai and Bajpai 1994; Bajpai
2001). Though these modifications reduce chlorinated compounds, but the major
drawback in most of these technologies is the ultimate disposal of sludge or con-
centrates which is more difficult and costly than the initial removal and separation.
This necessitates consideration of developing economical and eco-friendly methods
for removal of AOX compounds. Aerobic treatments are applicable where sufficient
molecular oxygen is available. The rate of degradation is proportional to dissolved
oxygen and therefore the process demands large inputs of energy, making it expen-
sive. Anaerobic treatment is a technically simple, relatively inexpensive technology
and consumes little energy. It also requires less space and produces less amount of
sludge. Anaerobic microorganisms can be preserved unfed for long periods of time
without any serious deterioration of their activity. The nutrient requirement for an-
aerobic treatment is low. It is less sensitive to toxic substances. Hence, it is proving
to be a viable technology for pulp and paper wastewater treatment. The major treat-
ment methods include anaerobic lagoon, anaerobic contact processes, Up-flow An-
aerobic Sludge Blanket (UASB), Sequencing Batch Reactors (SBR), fluidized bed,
anaerobic filters and hybrid processes. Now-a-days, high rate advanced anaerobic
reactors are being increasingly used. Anaerobic treatment remains the most reliable
and economically viable method of AOX removal at present.
Shooting ranges are of increasing environmental concern in many countries (Lin
1996; Mozafar et al. 2002; Sorvari 2007). High amounts of ammunition are often
deposited in the soils of these sites due to their use for shooting sports or military
activities. Depending on firing activities, considerable amounts of inorganic con-
taminants accumulate in shooting range soils and backstop materials. Due to the
alloys used for bullets and jacket housings lead, antimony, arsenic, bismuth, silver,
copper, and nickel may be present (Hardison et al. 2004; Johnson et al. 2005). An
environmental risk at shooting ranges has been determined from contamination lev-
els of groundwater and surface water (Sorvari et al. 2006; Heier et al. 2009), soil
enzymatic activity (Lee et al. 2002), and accumulation of heavy metals into plant
tissues (Labare et al. 2004), human being or other animals (Migliorini et al. 2004).
Current technologies for treating lead-contaminated soils mainly include solidifica-
tion/stabilization, vitrification, capping, secondary smelting, and soil washing.
Stabilisation of inorganic contaminants in soils is based on the modification of
pollutant characteristics (e.g. speciation, valence) and soil properties (sorption ca-
pacity, buffering potential, etc.) by means of additives (Diels et al. 2002). These
amendments induce or enhance physicochemical and/or microbial processes, which
render pollutants less mobile and less bioavailable. Due to specific interactions with
the constituents of the solid phase, cationic and anionic contaminants require differ-
ent additives. Cation exchange capacity may be increased by addition of synthetic
or natural clay minerals and iron oxides (McBride 1994; Lothenbach et al. 1999;
Bigham et al. 2002). An alternative approach involves the addition of soluble salts,
which provide anions to react with cationic contaminants forming leaching resistant
1 Environmental Protection Strategies: An Overview
18
minerals. A typical example is the addition of phosphate using commercially avail-
able phosphate fertilisers to stabilise heavy metals by precipitation of minerals with
low solubility like chloropyromorphite (McGowen et al. 2001) and thereby mini-
mise both plant uptake and leaching (Cao et al. 2002). Anion sorption capacity in
soils of the temperate zone is primarily controlled by iron(III)- and aluminium(III)-
(hydr)oxides like ferrihydrite, goethite, gibbsite, etc. Oxyanions like arsenate, chro-
mate, molybdate, etc. as well as cations like cadmium, copper, lead, and zinc are
sorbed specifically by these media (Richard and Bourg 1991; Bowell 1994; Marti-
nez and McBride 1999; Trivedi et al. 2003). The resulting inner sphere complexes
are resistant to competing anions/cations at typical levels in soil solutions. In ad-
dition, sorption may be accompanied by redox processes (Sun and Doner 1998)
leading to less toxic contaminant species. Following the reduction of mobility also
bioavailability can be expected to be reduced in stabilised soils.
In situ chemical immobilization is, in particular, a practical remediation technol-
ogy that is capable of reducing cost and environmental impacts (Saikia et al. 2006).
In situ chemical immobilization technologies can be employed in conjunction with
a plant application as a form of phytostabilization technology that stabilizes the soil
and prevents contaminant migration via wind and hydrological processes (Brown
et al. 2003; Mench et al. 2003). Kucharski et al. (2005) used calcium phosphate
to immobilize metal contaminants in soil with indigenous plant coverage that in-
creased water retention in the soil and reduced the volume of metal-containing
leachate. Use of plants in parallel with chemical immobilization technology is par-
ticularly important when the contaminated site is required to recover the vegetation
that has been degraded by metal toxicity.
Stabilization/solidification (S/S) is gaining prominence in the treatment and re-
mediation of hazardous wastes and contaminated soils due to its cost-effectiveness,
rapid implementation and its use of well-established techniques (Palomo and Pa-
lacios 2003; Dermatas and Meng 2003; Terzano et al. 2005). There are various
techniques currently used for S/S, including pozzolanic, (cementitious or solidi-
fying) based solidification systems and chemical based stabilization systems. The
most commonly applied pozzolanic materials are Portland cement, lime, and/or fly
ash (Dermatas and Meng 2003; Palomo and Palacios 2003; Terzano et al. 2005).
The pozzolanic-based S/S techniques immobilize contaminants by adsorption, in-
corporation into the pozzolanic products (e.g., calcium alumina hydrate C-A-H or
calcium silicate hydrate C-S-H), or via precipitation as metal hydroxides under al-
kaline pH associated with cement and lime (Gougar et al. 1996; Moulin et al. 1999).
Physical entrapment of heavy metals adsorbed to particle surfaces in a low perme-
ability cementitious matrix is also very likely (Moulin et al. 1999; Badreddine et al.
2004). Chemical-based S/S approach is based on the formation of thermodynami-
cally stable and insoluble precipitate end-products with the contaminants. More
effective chemical additives include phosphates and Fe-Mn oxides (Ma et al. 1995;
Hettiarachchi et al. 2000; Basta et al. 2001; Seaman et al. 2001; Cao et al. 2002;
Scheckel & Ryan 2004). The use of readily available and cost-advantageous materi-
als as immobilizing amendments becomes more significant when the remediation
targets vast amounts of contaminated soil such as shooting ranges.
A. Malik et al.
19
Dyes are an important class of pollutants, and can even be identified by the hu-
man eye. Disposal of dyes in precious water resources must be avoided, however,
and for that, various treatment technologies are in use. Treatment of synthetic dyes
in wastewater is a matter of great concern. Several physical and chemical methods
have been employed for the removal of dyes (Robinson et al. 2001). However, these
procedures have not been widely used due to high cost, formation of hazardous by
products and intensive energy requirement (Hai et al. 2007).
Among various methods adsorption occupies a prominent place in dye removal.
The growing demand for efficient and low-cost treatment methods and the impor-
tance of adsorption has given rise to low-cost alternative adsorbents (LCAs). Ex-
tensive research has been directed towards developing processes in which enzymes
are employed to remove dyes from polluted water (Bhunia et al. 2001; Shaffiqu
et al. 2002; Torres et al. 2003; Lopez et al. 2004; Husain 2006). Biological treatment
is the most common and widespread technique used in dye wastewater treatment
(Zhang et al. 1998; Bromley-Challenor et al. 2000; van der Zee and Villaverde
2005; Frijters et al. 2006; Barragan et al. 2007; dos Santos et al. 2007). A large num-
ber of species have been used for decolouration and mineralization of various dyes.
The methodology offers considerable advantages like being relatively inexpensive,
having low running costs and the end products of complete mineralization not being
toxic. The process can be aerobic, anaerobic or combined aerobic–anaerobic.
Bacteria and fungi are the two microorganisms groups that have been most wide-
ly studied for their ability to treat dye wastewaters. In aerobic conditions, enzymes
secreted by bacteria present in the wastewater break down the organic compounds.
The work to identify and isolate aerobic bacteria capable of degrading various dyes
has been going on since more than two decades (Rai et al. 2005). A number of
triphenylmethane dyes, have been found to be efficiently decolourized (92–100%)
by the strain Kurthia sp. (Sani and Banerjee 1999a). Nevertheless, it is worthwhile
pointing that synthetic dyes are not uniformly susceptible to decomposition by acti-
vated sludge in a conventional aerobic process (Husain 2006). Attempts to develop
aerobic bacterial strains for dye decolourization often resulted in a specific strain,
which showed a strict ability on a specific dye structure (Kulla 1981). Fungal strains
capable of decolourizing azo and triphenylmethane dyes have been studied in detail
by various workers (Bumpus and Brock 1988; Vasdev et al. 1995; Sani and Baner-
jee 1999b). Various factors like concentration of pollutants, dyestuff concentration,
initial pH and temperature of the effluent, affect the decolourisation process.
In order to get better remediation of coloured compounds from the textile ef-
fluents, a combination of aerobic and anaerobic treatment is suggested to give
encouraging results. An advantage of such system is the complete mineralization
which is often achieved due to the synergistic action of different organisms (Stolz
2001). Also, the reduction of the azo bond can be achieved under the reducing con-
ditions in anaerobic bioreactors (Brown and Laboureur 1983a) and the resulting
colourless aromatic amines may be mineralized under aerobic conditions (Brown
and Laboureur 1983b), thereby making the combined anaerobic–aerobic azo dye
treatment system attractive. Thus, an anaerobic decolourization followed by aerobic
1 Environmental Protection Strategies: An Overview
20
post treatment is generally recommended for treating dye wastewaters (Brown and
Hamburger 1987).
Oxidoreductive enzymes such as peroxidases and polyphenol oxidases are par-
ticipating in the degradation/removal of aromatic pollutants (Klibanov et al. 1983;
Dec and Bollag 1994). These enzymes can act on a broad range of substrates and
can also catalyze the degradation or removal of organic pollutants present in very
low concentration at the contaminated sites. In view of the potential of these en-
zymes in treating the phenolic compounds several microbial and plant peroxidases
and polyphenol oxidases have been considered for the treatment of dyes but none of
them has been exploited at the large scale due to low enzymatic activity in biologi-
cal materials and high cost of purification (Bhunia et al. 2001; Shaffiqu et al. 2002;
Verma and Madamwar 2002). The major reason that enzymatic treatments have not
yet been applied on an industrial scale is the huge volume of polluted wastewater
demanding remediation. Soluble enzymes suffer from certain drawbacks such as
thermal instability, susceptibility to attack by proteases, activity inhibition, etc. (Hu-
sain and Jan 2000). An important disadvantage of using soluble enzymes in the de-
toxification of hazardous aromatic pollutants is that the free enzyme cannot be used
in continuous processes. To overcome all these limitations enzyme immobilization
is the best alternative to exploit the enzymes at the industrial level.
The potential advantages of enzymatic treatment as compared to microbial treat-
ment are mainly associated to several factors; shorter treatment period; operation of
high and low concentrations of substrates; absence of delays associated with the lag
phase of biomass, reduction in sludge volume and ease of controlling the process
(Lopez et al. 2002; Akhtar and Husain 2006). However, the use of soluble enzymes
has some inherent limitations as compared to immobilized form of enzymes, which
has several advantages over the soluble enzymes such as enhanced stability, easier
product recovery and purification, protection of enzymes against denaturants, pro-
teolysis and reduced susceptibility to contamination (Husain and Jan 2000; Zille
et al. 2003; Matto and Husain 2006).
Numerous methods have been employed for the immobilization of peroxidases
from various sources but most of the immobilized enzyme preparations either use
commercially available enzyme or expensive supports, which increase the cost of
the processes (Norouzian 2003; Husain 2006). Such immobilized enzyme systems
cannot fulfill the requirements for the treatment of hazardous compounds coming
out of the industrial sites.
Besides, the known techniques used for the immobilization of enzyme, physical
adsorption on the basis of bioaffinity is useful as this process can immobilize en-
zyme directly from crude homogenate and thus avoid the high cost of purification.
The ease of immobilization, lack of chemical modification and usually accompany-
ing an enhancement in stability are some of the advantages offered by the adsorp-
tion procedures (Akhtar et al. 2005a, b; Kulshrestha and Husain 2006). Besides the
mentioned advantages offered by the bioaffinity-based procedures, there is an ad-
ditional benefit, such as proper orientation of enzyme on the support (Mislovicova
et al. 2000; Khan et al. 2005). These supports provide high yield and stable immo-
bilization of glycoenzymes/enzymes.
A. Malik et al.
21
Waste is an unavoidable by product of human activities. Rapid population
growth, urbanization and industrial growth have led to increase in the quantity and
complexity of generated waste and severe waste management problems in most
cities of third world countries. The large quantity of waste generated necessitates
a system of collection, transportation and disposal. It requires knowledge of what
the wastes are comprised of, and how they need to be collected and disposed. Sol-
id waste in general, comprises of municipal solid waste (MSW) which includes
household and commercial wastes; agricultural waste; and non-hazardous industrial
waste; and construction and demolition waste.When solid waste is disposed off on
land in open dumps or in improperly designed landfills (e.g. in low lying areas),
it causes the following impact on the environment: ground water contamination
by the leachate generated by the waste dump, surface water contamination by the
run-off from the waste dump, bad odour, pests, rodents and wind-blown litter in and
around the waste dump, generation of inflammable gas (e.g. methane) within the
waste dump. Some commonly used methods by which the waste could be managed
are: land filling, incineration, composting and recycling.
Municipal Solid Waste Management involves the application of the principle of
Integrated Solid Waste Management (ISWM) to municipal waste. ISWM is the ap-
plication of suitable techniques, technologies and management programs covering
all types of solid wastes from all sources to achieve the twin objectives of (a) waste
reduction and (b) effective management of waste still produced after waste reduc-
tion. An effective system of solid waste management must be both environmentally
and economically sustainable.
Solid waste incineration is another method of waste management. Trash is put
into large incinerators which convert it into steam, gas, heat and ash. This process
can sometimes be time intensive. However, it is effective in disposing hazardous
waste. The most significant negative outcome of incineration is the emissions that
result from combustion. This air pollution has both a harmful effect on the local
area and on the planets climate (Buonanno et al. 2008). Greenhouse gas emissions
(GHGs) mainly in the form of CO
2
and N
2
O are the main contributors to climate
change through incineration (Gutierrez et al. 2005).
Recycling involves (a) the separation and sorting of waste materials; (b) the
preparation of these materials for reuse or reprocessing; and (c) the reuse and repro-
cessing of these materials. Recycling is an important factor which helps to reduce
the demand on resources and the amount of waste requiring disposal by landfilling.
Recycling of waste proves to be an effective management option because it does
not involve the emission of many greenhouse gases and water pollutants. Aside
from the traditional methods of waste management, biowaste has been used in the
production of clean energy where it replaces coal, oil or natural gases to generate
electricity through combustion. This waste-to-energy conversion process has been
proved to be safe, environment friendly and reduces the incoming volume by 90%;
the remaining ash is used as a roadbed material or as a landfill material.
Landfilling involves the controlled disposal of wastes on or in the earth’s mantle.
Landfills are used to dispose of solid waste that cannot be recycled and is of no
further use, the residual matter remaining after solid wastes have been pre-sorted
1 Environmental Protection Strategies: An Overview
22
at a materials recovery facility and the residual matter remaining after the recovery
of conversion products or energy. It is by far the most common method of ultimate
disposal for waste residuals. Many countries use uninhabited land, quarries, mines
and pits as landfill sites. Biological reprocessing methods like composting and an-
aerobic digestion are natural ways to decompose solid organic waste. Composting
is nature’s way of recycling organic wastes. Composting is a method of decompos-
ing waste for desposal by microorganisms (mainly bacteria and fungi) to produce
a humus-like substance that can be used as a fertilizer. This process converts waste
which is organic in nature to inorganic materials that can be returned to the soil as
fertilizer i.e. biological stabilization of organic material in such a manner that most
of the nutrient and humus that are so necessary for plant growth are returned to the
soil.
Health issues are associated with every step of the handling, treatment and dis-
posal of waste, both directly (via recovery and recycling activities or other occupa-
tions in the waste management industry, by exposure to hazardous substances in the
waste or to emissions from incinerators and landfill sites, vermin, odours and noise)
or indirectly (e.g. via ingestion of contaminated water, soil and food). The main
pathways of exposure are inhalation (especially due to emissions from incinerators
and landfills), consumption of water (in the case of water supplies contaminated
with landfill leachate), the foodchain (especially consumption of food contaminated
with bacteria and viruses from landspreading of sewage and manure, and food en-
riched with persistent organic chemicals that may be released from incinerators). It
is also important to remember that occupational accidents in the waste management
industry can be relatively common, higher than national average for other occupa-
tions (HSE 2004), and often higher than the potential cases of adverse effects to
the resident population investigated by epidemiological studies. The main cause
of global warming is the increasing amount of greenhouse gases (CO
2
, CH
4
and
N
2
O) in the atmosphere, a significant contribution comes from waste management
practices (Smith et al. 2001).
Agriculture and forests form an important resource to sustain global economical,
environmental and social system. Their protection against pests is a priority and
due to the adverse impact of chemical insecticides, use of biopesticides is increas-
ing (Marrone 1999). A number of biopesticides (bacteria, fungi, virus, pheromones,
plant extracts) have been already in use to control various types of insects respon-
sible for the destruction of forests and agricultural crops. Bacillus thuringiensis
(Bt) based biopesticides are of utmost importance and occupy almost 97% of the
world biopesticide market (Cannon 1993). A biological pesticide is effective only
if it has a potential major impact on the target pest, market size, variability of field
performance, cost effectiveness, end-user feedback and a number of technological
challenges namely, fermentation, formulation and delivery systems (Jacobsen and
Backman 1993; Copping 1998). Development cost, time and ease of registration
and potential growing market in contrast to chemical pesticides make biopesticides
interesting proponents to investigate. Despite, extensive research in the field of Bt
biopesticides, many formulations do not deliver effectively in field owing to variable
environmental stress (for example, forestry and agriculture). Another reason could
A. Malik et al.
23
be adoption of integrated approach which can play an important role in biopesticide
development, in other words, tailoring fermentation and harvesting processes to
produce higher potency efficacious formulations. Biopesticide research has been
comprehensively detailed in Burges (1998), but there are recent advances, which
have taken place henceforth. Meanwhile, wastewater (WW) and wastewater sludge
(WWS) based Bt formulations also need to be elaborated and discussed due to their
inherent positive features.
The Environmental Protection Strategy is entirely based on the principles of sus-
tainable development. The uprising population and the environmental deterioration
face the challenge of sustainable development. The remedy of environmental prob-
lems requires great financial resources, for instance, the remediation in the solid
waste and wastewaters sectors requires huge investments. Bioremediation uses rela-
tively low-cost, low-technology techniques, which generally have a high public ac-
ceptance. The treatment of aqueous and solid wastes of industrial, agricultural and
domestic origin offers a number of opportunities to apply a wide range of biotech-
nological methods. The most essential resources for food production are water, soil
and energy. Biotreatment and bioremediation techniques are useful tools to control
water quality, monitor pollution, decontaminate wastewaters and prevent pollution.
Bioremediation seems to be a good alternative to conventional clean-up technolo-
gies and research in this field is rapidly increasing. When used as a component of
Integrated Pest Management (IPM) programs, biopesticides can greatly decrease
the use of conventional pesticides, while crop yields remain high and the pollution
problems caused by conventional pesticides are avoided. The potential of rhizo-
spheric microorganisms like mycorrhizal fungi and rhizobacteria, which contribute
essentially to increase the soil fertility and remediate physically and chemically
disturbed soils should be utilised. The major advances in molecular methodologies
that have been achieved in the recent past will provide improved in vivo models for
mutagenecity and genotoxicity testing in the near future. The potential of enzymes
in cleaning up wastes and biodegradation of contaminants might be even greater as
a result of “directed evolution”, which has led to the production of highly efficient
enzymes.
The various technologies summarized in this chapter could play a major role in
most of these fields but will they, in all situations, be efficient and effective enough
to justify the necessary investment? A critical evaluation of current approaches and
results is needed in order to determine this.
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