Keywords

9.1 Introduction

The building blocks of nanotechnology are nanoparticles (Biswas and Wu 2005). The term nanoparticles is derived from the word ‘nanos’ meaning tiny. Particles that are measured in nanoscale level, i.e. 0.1–100 nm in diameter, are termed as nanoparticles (Royal society and Royal Academy of Engineering 2004). They have been present on earth and used by mankind for millions of years (Nowack and Bucheli 2007). Nanotechnology is defined as the design, synthesis and application of materials, devices whose size and shape have been engineered at the nanoscale (Buzea et al. 2007). It has bridged the gap among various other existing technologies of present scenario. It has mingled chemistry, physics, biology, engineering and technology, bioinformatics and biotechnology in such a manner that has offered a novel concept of interdisciplinary research. Nanoparticles play a key role and are known as the backbone of this technology due to their distinguishing and incomparable properties. The small size offers a large surface area as compared to their bulk counter parts (Biswas and Wu 2005). The large surface to volume ratio, surface charge, geometry and state of existence decide their relative reactivity.

Nanoparticles are broadly classified as natural nanoparticles and manmade (anthropogenic) nanoparticles. Anthropogenic nanoparticles are further sub divided into unintentional and intentional/engineered nanoparticles. A variety of nanoparticles are introduced in the environment by various natural processes. Volcanic eruptions, forest fires, weathering phenomenon and soil erosion driven by water and wind are the sources of natural nanoparticles (Smita et al. 2012). Whereas, biological origin i.e. viral particles (Hogan et al. 2004), Pollen fragments (McMurry et al. 2000). Coal fired combustion systems and incinerators (Chang et al. 2004); automobiles and diesel powered vehicles (Kittelson 1998); welding processes industrial boilers, fire places, automobiles, diesel (Vincent and Clement 2000); trucks, and meat-cooking operations (Hildemann et al. 1991) are the sources of unintentional anthropogenic nanoparticles. The intentional/engineered nanoparticle i.e. carbon containing nanoparticles (fullerence, carbon nanotubes etc.) inorganic nanoparticles (metal nanoparticles like nano silver, nano gold, titanium dioxide nanoparticles, zinc oxide nanoparticles etc.), organic and inorganic nanohybrids (Sajid et al. 2015) are synthesized by following two approaches i.e. either top down or bottom up method.

The engineered nanoparticles have been exploited in every sector of society. They have become a compulsive content of today’s smart products. Their rapidly increasing application in textiles, electronics, pharmaceutics, cosmetics and environmental remediation is noticeable and calls for their impact assessment (Royal society and Royal Academy of Engineering 2004). Table 9.1 corroborates the application of different nanoforms.

Table 9.1 Application of nanoparticles

In the present review we have taken an attempt to commemorate the toxicity imposed by the nanoparticles on every sphere of life forms. Brief information about toxic effect of nanoparticles on the environment, microbes, animals and plants has been discussed in this article. The possible preventive measures and future challenges are also mentioned.

9.2 Nanotechnology, an Old Concept

The concept of nanotechnology was proposed early in 1959 by a physicist Richard Feynman in a lecture-“There’s Plenty of Room at the Bottom” at a meeting of the American Physical Society and was honored by Nobel prize for this valuable contribution. Whereas, the term nanotechnology was coined by Norio Taniguchi in 1974. Unknowingly, the people were using the nanoparticles in their work place. Evidences of nanotechnology’s existence in the middle ages are drawn from the stained glasses of various old churches. The glass windows of European cathedrals are live examples of this. The historical aspect of nanotechnology from the most primitive time to the middle age was commemorated by (Daniel and Astruc 2004). Due to very astonishing optical properties of nanoparticles they have been used since the fourth century, the prominent example of which is the dichroic glass of Lycurgus cup whereas, the use of nanomaterials in industrial sectors begins in twentieth century (Horikoshi and Serpone 2013). Use of silver nanoparticles as an antiseptic agent has been documented early in Rig veda. Various metals in nano form occupies a remarkable place in ayurvedic, sidhha and unani system of medicine due to their terrific potential in curing a wide range of diseases and abnormalities.

9.3 Distinguishing Properties of Nanoparticles

Before highlighting the hazardousness of nanoparticles the expected factors that are responsible for nanoparticle toxicity are size, shape, nature, reactivity, mobility, stability and surface chemical charge (Sajid et al. 2015). Property of the material changes in nanoparticle regime. Activity of the nanoparticles are strongly governed by their stability which is attained by the provision of ideal capping agents. Temperature, pressure and long term incubation governs the texture and consistency of the nano regime material due to the onset of aggregation or agglomeration among the nanopopulation.

9.4 Effect of Nanoparticles on the Environment

Environment harbors all the things that exist within itself. Any alteration in the environment gradually affects its state, constituents and vice-Versa. Environment consists of two factors broadly classified as biotic and abiotic factor. Nanoparticles that are expelled in the environment either naturally or by anthropogenic means, Spontaneously through natural phenomenon or by engineered sources imparts/leads to some changes in them. The inhalation of atmospheric nanoparticle has caused 60,000 deaths per year reported by US Environmental Protection Agency (EPA) and also these Nanoparticles directly get transferred in to the brain Oberdorster et al. (2004) and Raloff (2003). This is due to the large surface area which has resulted in direct generation of reactive oxygen species that are known as harmful oxyradicals, having the potential to attack DNA, proteins and membranes too as a result of which the cell gets injured Brown et al. (2001). In addition they show affinity towards the transition metals and organic chemical pollutants that enhances the level of toxicity to a greater extent Cheng et al. (2004) Route of entry of nanoparticles varies with the type of habitat. Figure 9.1 reflects the possible route of nanoparticle exposure to various life form in brief. In terrestrial organisms they may enter via inhalation or ingestion Brigger et al. (2002) whereas, in case of the aquatic animals they enter via direct passage across gill and other external surface epithelia as reported by Moore (2006). At cellular level engineered nanoparticles (single wall nanotubes) are responsible for swelling of the endoplasmic reticulum, it causes vacuolar changes. Additionally, high concentration of multiwalled nanotubes were found to be responsible for the degeneration, enlargement and rarefaction of macrophage cell’s nucleus Jia et al. (2005).

Fig. 9.1
figure 1

Showing the sources of nanoparticles and their impact on various components of ecosystem

In a nutshell, this can be clearly stated that nanoparticles (1–100 nm) can be life threatening. Source may vary but once entered in the environment they may harm every level of ecosystem.

9.4.1 Effect of Nanoparticles on Atmosphere

Accumulation of nanoparticle in the environment leads to generation of various environmentally toxic processes like dust cloud formation, Soot formation (Regional haze). These processes occur as a result of burning of organic waste materials found in nature that has been responsible for melting of Himalayan Glaciers Gustafsson et al. (2009). These are actually the naturally occurring nanoparticles that interfere with the natural resources. Incomplete combustion of fossil fuels increases risk of exposure to pathogenic species as suggested by Hua et al. (2007). According to Manning et al. (2005) others ill effects include abnormalities in level of environmental hydroxyl radical concentration. Wilson et al. (22007) reported the increase in ozone depletion mechanism by these tiny creatures that may be attributed due to change in tempaerature of stratospheric layer as reported by Tromp et al. (2003), Smita et al. (2012). According to Strawa et al. 2010 nanoparticles take part in the coarse material formation which is the major cause of global warming in present scenario.

9.4.2 Effect on Soil

The ultimate sink of every component in environment is soil and so as for nanoparticles. These tiny tools end up their journey in the soil or become dormant for some time. Soil is a natural reservoir for a wide variety of micro flora as it offer conducive environment for growth and survival of biological indicators. In turn they offer nutrients in the form of enzyme that take part in decomposition of soil content and improve the quality of soil by executing this cyclic process. In a broad sense their exists mutualism. The long term dormancy period of nanoparticles may evoke serious issues on the health of its inhabitant i.e. contribute in reducing the fertility of soil thereby causing an imbalance in the soil ecosystem (Kennedy and Smith 1995). Common ENPs include the metal ENPs (elemental Ag, Au, Fe etc.), fullerenes (grouping Buckminster fullerenes, CNTs, nanocones etc.), metal oxides (TiO2, CuO, FeO2, ZnO, Al2O3, SiO2 etc.), complex compounds (Co–Zn–Fe oxide), quantum dots often coated with a polymer e.g. cadmium–selenide (CdSe) and organic polymers (dendrimers, polystyrene, etc.) Dinesh et al. 2012. Fortner et al. (2005) performed toxicity study of C60 aggregates on soil inhabiting E. Coli and B. subtilis bacteria. They found it to be toxic even at low concentration marked by reduced growth (>0.4 ppm). more over, this concentration has lowered the rate of aerobic respiration up to 4 ppm. Thus may interfere with soil development (pedogenesis) or behavior (e.g., crusting and dispersion) Klaine et al. (2008), Cameron et al. (1915).

9.4.3 Effect of Nanoparticle on Water

Domestic and Industrial wastes (may contain Nanoparticles in aggregate or colloidal form) expelled in the water bodies viz. ponds, rivers, sea etc. increases the threat of nanoparticle contamination. Discharges usually contains organic (protenaceous and polysaccharide from the microbes) and inorganic matter (heavy metal oxides) that make the water bodies unfit for use. It has been suggested that Ionic silver has found to be highly toxic that disrupts Na+K+− ATPase enzyme activity due to which active Na+ and Cl− uptake is blocked that causes Osmoregulation by fishes, leads to death of fishes thereby contaminate the water as studied by Tang and Wang (2004). Nanoparticles may also accumulate on the surface microlayers of the oceans, presenting a route of aerosol exposure risk to marine birds and mammals, as well as the organisms living on the surface microlayer due to their hydrophobic nature. Kennedy et al. (2004), Simkiss K. (1990).

9.4.4 Effect of Nanoparticles on Food Chain

Starting from the primary producer (Plants) to that of the tertiary consumers (insects/animals). At every successive level nano-particles reflects their involvement by being entered in their metabolic pathway through active and passive transport. Confirmatory reports have been published by Holbrook et al. (2008), Lin et al. (2009) that shows their harmful effect in the food chain. Their small size offers large surface area that imparts them high potential to carry toxic materials i.e. such as lipophilic pollutants and heavy metals Baun et al. (2008) to enter the system.

9.4.5 Effect of Nanoparticles on Agriculture Production

About 70 % of India’s economy is based on agricultural practices. The formation of dust clouds has found to be highly hazardous. Burning of wood for cooking and other purposes and the incomplete combustion of biomass give rise to natural nanoparticle. Their existence in environment has resulted in brown cloud formation which has remarkably lowered the sunlight intensity in south Asia. These pollution clouds have been indulged in the reduction of monsoon season in India. The productivity of chief crop plants like rice, wheat and soybean has immensely hampered as reported by UNEP 2002. Asian brown clouds are found to be loaded with large amount of soot and black carbon. These complexly degradable materials contributes to increase in melting of glaciers thereby blocking the availability of water to various glacier fed rivers as commemorated by Gustafsson et al. (2009). Water scarcity contributes in low yield of food crops.

9.5 Effect of Nanoparticles on Microbes

Nanoparticles not only affected abiotic factors rather have also been observed to pose harmful effects on microscopic creatures too ranging from the prokaryotes to Eukaryotes. For example – Akhavan and Ghaderi (2010) investigated the toxicity of graphene and graphene oxide damages the cell membrane of both the Gram-negative bacteria Escherichia coli and the Gram-positive bacteria Staphylococcus aureus by their sharp edges. A long descriptive about nanoparticles impact on micro flora has been expressed in Table 9.2.

Table 9.2 Harmful effect of nanoparticles on microorganisms

9.5.1 Effect of Nanoparticles on Nitrogen Fixing Bacteria (Cyanobacteria & Others)

Cyanobacteria are integral part of producer community as they are the largest nitrogen fixer globallydue to the presence of nif gene and other ideal characteristics they are capable of fixing the atmospheric nitrogen themselves and serves as Biofertilizers thereby play a significant role in the growth of greenery. Due to the presence of oxygenic photosynthetic system they are capable of maintaining the equilibrium of earth atmosphere. These certain properties drag them in the list of essentials that vitally contribute for balancing the environment and nature. Brayner et al. (2010) performed the eco-toxicological investigation of ZnO Nanoparticles by taking Anabaena flos-aquae cyanobacteria and Euglena gracilis euglenoid microalgae as the experimental organism. ZnO Nanoparticles were synthesized by polyol process in which s trin-octylphosphine oxide (TOPO) and polyoxyethylene stearyl ether (Brij-76) were used as protective agents to control the particle size and shape. In the present investigation effect of ZnO nanoparticles on photosynthetic activity of the selected organisms was corroborated. In the case of Anabaena flos-aquae, addition of ZnO, ZnO-TOPO, and ZnO-Brij-76 induces stress which causes a progressive decline in the photosynthetic rate. Among these three different types of nanoparticles only ZnO-TOPO causes a fall in photosynthetic rate followed by cell death. However, In case of Euglena gracilis, all the nanoparticles were found to be indulged in cell death which was confirmed by doing the TEM analysis of their ultra thin sections. It reveals that polysaccharides produced by Anabaena flos-aquae avoid particle internalization after contact with ZnO and ZnO-Brij- 76 nanoparticles. Ma et al. (2013) suggested that nanoparticles have been found to interfere in the metabolic pathway of nitrogen fixing bacteria as well.

9.5.2 Effect of Nanoparticles on Plant Growth Promoting Rhizobacteria

Plant growth promoting rhizobacteria (PGPR) like P. aeruginosa, P. putida, P. fluorescens, B. subtilis and soil Nitrogen cycle bacteria viz., nitrifying bacteria and denitrifying bacteria have shown varying degrees of inhibition when exposed to ENPs in pure culture conditions or aqueous suspensions Mishra and Kumar (2009). Metal oxide nanoparticles of Cu (80–160 nm) were tested for antibacterial activity against plant growth promoting Klebsiella pneumoniae, P. aeruginosa, Salmonella paratyphi and Shigella strains Mahapatra et al. (2008). Iron and copper based nanoparticles are presumed to react with peroxides present in the environment generating free radicals known to be highly toxic to microorganisms like P. aeruginosa Saliba et al. (2006).

9.5.3 Effect of Nanoparticles on Eukaryotes

Likewise, the prokaryotes nanoparticles adversely affects the growth and metabolism of eukaryotes. Kasemets et al. (2009) has shown the toxic effect of ZnO nanoparticles on the unicellular Fungi i. e. yeast (Saccharomyces cerevisiae). Mortimer et al. (2010) suggested the toxicity of ZnO nanoparticles against free living Protozoan Sp. (Tetrahymena thermophila).

9.6 Effect of Nanoparticles on Plants

9.6.1 Effect of Nanoparticles on Algae & Diatom (Lower Plants)

Due to the adsorption of TiO2 nanoparticles on the surface of algae, its weight becomes double as a result of this its floating capacity decreases and low surface gets exposed to sunlight leads to a gradual fall in photosynthetic rate as suggested by Navarro et al. (2008). In case of a green algae i.e. Desmodesmus subspicatus, TiO2 nanoparticles were found to be toxic by Hund-Rinke and Simon (2006). ZnO nanoparticles showed toxicity against Euglena gracilis Euglinoid Brayner et al. (2010). Marine diatom T. pseudonana was found to be susceptible to ZnO nanoparticles and exhibit toxicity as reported by Miao et al. (2010).

9.6.2 Effect of Nanoparticles on Marine Phytoplanktons

Population growth studies on four marine phytoplanktons has been conducted by Miller et al. (2010) to assess the toxicity estimation of two metal oxide nanoparticle i.e. ZnO and TiO2. Salty water bodies comprises a vast portion of earth and a repeated history of waste disposition has been in a common practice since ancient time, whether by accidental or intentional means. Marine phytoplanktons are adversely affected by engineered nanoparticles. In order to assess the sublethal effect of two selected metal NPs a modeling approach (Dynamic Energy Budget (DEB) framework) was employed in which They have found that ZnO nanoparticles depressed the growth rate of Skeietonema marioni, Thalassiosira pseudonana, Dunaliella tertiolecta and Isochrysis galbana by 50–70 %. Reproduction and early stage developmental study has been conducted by Nielsen et al. (2008). They have selected a marine brown algae Fucus serratus to test against manufactured Carbon black nanoparticle. Inference clearly states a great changes. Carbon black nanoparticles when subjected to the sperm suspension and embryo culture gets agglomerate thereby causing a decline in the fertilization success at a concentration of 100 mgl−1.

9.7 Effect of Nanoparticles on Higher Plants

Nanoparticles are found to be indulged in the formation of reactive oxygen species in some higher plants explained by Navarro et al. (2008). Accumulation of nanoparticle takes place on the leaf lamina and on stomatal tissues causes an imbalance in the equilibrium inside outside the leaf and ultimately in plants Da Silva et al. (2006). Accumulation of nanoparticle on photosynthetic surfaces causes shading effects, i.e. reduced sun light availability and hence reduced photosynthetic rate Smita et al. (2012). Carbon nanotubes diminished rice yields and made wheat more prone to other pollutants Wild and Jones (2009) due to the asbestos-like behaviour of carbon nanotubes. Also, aluminium nanoparticle have been identified to inhibit root growth in various economically important plant species by interacting with root surface Canas et al. (2008), Yang and Watts (2005).

9.7.1 Phytotoxicity by Nanoparticles

Y. K. Mushtaq (2011) demonstrated The effects of Fe3O4, TiO2, and carbon nanoparticles on cucumber plants. These nanoparticles exhibits the potential to negatively affect seed germination rate, root elongation, and germination index. Another study performed by Kumari et al. (2011) on the harsh effect of ZnO nanoparticles supports their toxic nature. Growing use of ZnO nanoparticles in the daily consumer products calls for an alert after their significant investigation. Workers have done a depth study by determining the Mitotic index (MI), micronuclei index (MN index), chromosomal aberration index, and lipid peroxidation of the root cells of A. cepa (grown hydroponically). They have treated the model organism with four different concentrations of ZnO nanoparticles dispersions (25, 50, 75, and 100 gml−1). It was observed that MI decreases with the increase in concentration of ZnO nanoparticles to the contrary MN index and chromosomal index increases steadily. in the ZnO nanoparticles treated cells, the total count of micronucleated cell were higher than control set up which corroborates the genotoxic effect of ZnO nanoparticles on plant community.

9.7.2 Genotoxicity by Nanoparticles

Atha and collaborators (2012) reported for the first time that copper oxide Nanoparticles damaged DNA in some agricultural and grassland plants (Raphanus sativus, Lolium perenne, and Lolium rigidum). Mutation occurs in the crop plants that blocks their growth and results in gradual fall in productivity. Through this fraction of investigation they have clearified that metal nanoparticles acts as a carrier/mediator for DNA damage in all the living community including mammalian cell, plants and even the microscopic beings i.e. bacteria (Table 9.3).

Table 9.3 Harmful effect of nanoparticles on plants

9.8 Effect of Nanoparticles on Aquatic Organisms

To assess the merits and demerits of engineered nanoparticles several comparative studies has been done since time immemorial. These documentation provide a thorough and in depth study of the other face of a coin say the negative impacts of nanomaterials. Nanoparticles are being exploited and used in diversified applications. And being a multifunctional paradigm researchers have warned to use this versatile tool in an ecofriendly manner. Deleterious effects of nanoparticles are being experienced by atmosphere and land than how could the water bodies that covers 71 % of the earth surface will be ignorant and its inhabitant as well carbon nanotubes and fullerenes found to be potentially harmful especially to benthic organisms Velzeboer et al. (2011). Several researchers have excavated the harmful effects of nanomaterials by following different types of approaches. In recent years Blinova et al. (2010), have performed a comparative analysis of acute toxicity test of CuO and ZnO nanoparticles between crustaceans and protozoan population found in an artificially prepared fresh water ecosystem and a natural water body. In the study acute toxicity of crustaceans Daphnia magna and Thamnocephalus platyurus and protozoan Tetrahymena thermophila was determined.

9.9 Effect of Nanoparticles on Terrestrial Organisms

9.9.1 Effect of Nanoparticles on Terrestrial Invertebrates

Invertebrates cover a large group of animals ranging from top to bottom trophic levels of ecosystem. They are involved in various physiological phenomenon i.e. pollination, soil formation etc. Their occurrence differ from one habitat to the other and are considered as bioindicators (symbol of environmental change). Being an integral part of environment obviously this group of organisms encounters with the nanoparticles. In this section authors have tried to reflect the effect of nanoparticles on Annelids.

9.9.2 Effect of Nanoparticles on Annelids

Agricultural practices are the most important and widely used occupation cum source of income world wide. In order to earn the livelihood and growing demand of requirements one has to look for an effective and environmentally safe tool to reach the target of high productivity without compromising with the chemical based fertilizers and pesticides that has proved to be indulged in the formation of waste lands by successively decreasing the humus and fertility rate of the soil ultimately leading to a major cause of Bioaccumulation and biomagnification. Organic farming is the best alternative in which earthworms play the most significant role. Keeping in view the immense importance of earthworms in agriculture and zooming application of engineered nanoparticles in almost every sector of society studies has been conducted to see the effect of TiO2 and ZnO nanoparticles by growing earthworms Eisenia fetida in an artificially created soil system by Hu et al. (2010). Distilled water at a concentration of, 0.1, 0.5, 1.0 or 5.0 g kg1 of Nanoparticles were prepared and earthworms were placed in that soil system for 7 days. By using Acute toxicity test and Comet assay concentration of Ti and Zn in earthworms was determined. It was observed that when doses of TiO2 and ZnO were greater than 1.0 g kg−1 Zn accumulated in higher concentration than Ti and this leads to hyperaccumulation of Zn in the mitochondria of its gut cells which causes DNA damage. The cellulose activity was also inhibited at the dose level of 5.0 g kg1 of ZnO nanoparticles. Likewise, Khare et al. (2011) reported the toxicity of TiO2 and ZnO nanoparticles less than 25 nm and 100 nm on the nematode (Caenorhabditis elegans). In this study toxicity of small and large sized nanoparticles of ZnO and TiO2 were compared. It was found that small sized (25 nm) ZnO nanoparticles. were more toxic than that of the small (25 nm) and large sized (100 nm) TiO2 nanoparticles.

9.9.3 Effect of Nanoparticles on Vertebrates

In the previous sections an attempt has been made to throw light upon the effect of nanoparticles on various abiotic and biotic factors of the environment. In this section the hazardous effect of inorganic nanoparticles i.e. ZnO has been demonstrated on two groups of vertebrates i.e. amphibians and mammals.

9.9.4 Effect of Nanoparticles on Amphibians

ZnO nanoparticles brought about developmental abnormalities in amphibians. Metallic nanoparticles have been found to cause deleterious effect up on encountered life forms. Among them ZnO, TiO2, Fe2O3, and CuO nanomaterials (20–100 nm) were subjected to access their effect on amphibians in a study conducted by Nations et al. (2011a) on Xenopus laevis by employing the Frog Embryo Teratogenesis Assay Xenopus (FETAX) protocol. Experimental organism was subjected to dispersed nanomaterials at a concentration upto 1000 mg L−1 for TiO2, Fe2O3, CuO, and ZnO. It was observed that inspite of causing mortality to the test specie these causes developmental abnormalities among them Viz.gastrointestinal, spinal, and other abnormalities. However, at a concentration of 10.3 mg L−1 of ZnO leads to total malformation. This study confers the harmful effects of ZnO nanoparticles. so its use and production needs to be checked for future perspective.

9.9.5 Effect of Nanoparticles on Mammals

Nanoparticles have been found to cause a number of serious problems such as cytotoxicity. Copper nanoparticles while incubated with zinc ions remains normal but when exposed to large sized ZnO nanoparticles. (at a non toxic concentration 6.25 μg/ml) its cytotoxic activity enhances gradually, also synergistic activity becomes strong. This is due to the presence of large sized ZnO nanoparticles which brings about the alteration in the human hepatoma cell line HepG2 Li et al. (2015) Table 9.4.

Table 9.4 Harmful effect of nanoparticles on animals

9.10 Conclusion

Nanoparticles are the most promising and widely exploited resources of present time. Their surface properties makes them unique, thus they are being used in every sector of society. To make and prove this technology as a boon we need to take some simple measures. By taking care while handling the engineered nanomaterials, by properlystudying the fate of the synthesized nanoparticle, also by implementing ecofriendly techniques that consumes minimum energy, labour and cost effective methods we can add beauty to our living world inspite of accelerating the pollution rate. Use of chemically engineered nanoparticles should be minimizes. Establishment of regulatory guidelines for nanoparticle synthesis is the urgent call, which should be followed by both the developed and developing nations. Now it’s the need of time that without delaying the slogan of “GO Green” should be practiced not in words but in work. One thing is to be kept in mind i.e. the life cycle of engineered material should be well studied and the large scale synthesis of them should be executed on the basis of their fate in the environment. In the quest of innovation and advancement environmental issues are becoming more serious and uncontrolled. So, let’s shake a leg towards the betterment of the earth in which we are living in.