eChapters

ABSTRACT

An understanding of the metabolism of heavy metals facilitates biotechnological use of these functions just recently, although heavy metals comprise the major part of the elements in the periodic table. Due to their ability to form complex compounds, some heavy metals are essential trace elements. In any case, essential or not, most heavy metals are toxic at higher concentration. Heavy metal pollution from last few decades has led to an increased environmental concern. Heavy metal like cadmium is a non-essential toxic heavy metal and is also being increased in soil during last few years posing a serious threat to environment and human health worldwide. Therefore, remediation strategies of the contaminated sites are great concerns and can be achieved by a number of different physico-chemical and biological means.

1. Introduction
Agro-industrial waste plays a crucial role in bioremediation, the process of usingbiological organisms to remove or neutralize pollutants from contaminatedenvironments (Garg, 2020). With the rise of industrialization and agriculturalactivities, there’s a corresponding increase in waste production. However,these waste materials, often rich in organic compounds, can be repurposedas valuable resources as mentioned in scientific studies (Sahota and Sharma,2020; Yadav et al., 2022).

1. Introduction to Biomedical Waste (BMW)
The waste produced at different medical facilities, including hospitals,laboratories, research centers, mortuaries, and autopsy centers is considered asBiomedical Waste (BMW). Some of the BMW is hazardous whereas some arenon-hazardous in nature. Hazardous waste includes, Infectious, Pathological,Microbiological, Pharmaceutical, Chemical, Cytotoxic and Radioactive wastematerial as described by World Health Organization (Fig. 1). Healthcarepersonnel and the general public are at risk from physical, chemical, andmicrobiological effects of hazardous waste. On the other hand, the nonhazardousBMW which are not harmful, include packaging material made upof plastic & thermocol, cardboards, paper waste etc. (Capoor & Parida, 2021).

1. Introduction
Since plastic was discovered, global plastic production for industrial scale usages has grown exponentially, causing a tremendous increase in plastic waste from a few tons in the 1950's to almost 400 million tons in 2018 (Syberg et al. 2021). Plastic is non-biodegradable and ubiquitous.

Environmental degradation of plastic operates through a mechanism ofphysical fragmentation, photodegradation and microbial degradation (Wright
et al., 2020), resulting in fragmentation of larger plastics into smaller plasticparticles, also called microplastics and nanoplastics (MNPs). These debris areserious environmental threats (Hartmann et al., 2019).

Abstract

Excessive use and treatment of soil with pesticides under the adage, “if little is good, a lot more will be better” has played havoc with human and other life forms and can also cause populations of beneficial soil microorganisms to decline, which indirectly affect soil fertility. Soil constitutes a major environmental sink for many pesticides. These pesticides are either taken up from soil by plants and then pass into the bodies of invertebrates, water or air, directly or indirectly and affect public health and beneficial biota of the ecosystem. Bioremediation provides very attractive, eco-friendly and economic solution to many of our hazardous pollution problems. For both economic and ecological reasons, biological degradation has become an increasingly popular alternative for treatment of hazardous wastes.

1. Introduction
Plastic pollution is a global problem that requires immediate attention due toits negative impacts on ecosystems, infrastructure, (MacLeod, M., Arp et al.2021) society, and the economy (Honingh, D. et al.,2020).In terms of absolute emissions, we find that South-eastern part of Asia, andSub-Saharan part of Africa possess the greatest quantities of plastic pollution,with India emitting the most (9.3 Mt year−1), or about one-fifth of the world'stotal plastic pollution (Joshua W. Cottom et al.).
Future regulations pertaining to the manufacturer, usage, and getting ridof plastics will be influenced by the discussions for a worldwide plasticpollution treaty. A baseline with great resolution of waste flows and sourcesof plastic emissions will be useful for the stakeholders in order to identify
pollution hotspots and the reasons behind them (United Nations EnvironmentProgramme). Furthermore, in addition to the production of municipal waste,the pandemic of coronavirus illness has raised the usage of disposable polymerproducts, masks, tissues, gloves, and additional personal protection equipment,which made the plastic pollution disaster worse (De-la-Torre GE et al., 2021).Wasted plastics can break down and fragment into three different sizes: 1mm to 5 mm big microplastics, 1 μm to 1 mm microplastics, and 1 nm to1 μm nano plastics, (Waller et al., 2017; De-la-Torre 2020; Atugoda et al.,
2022) being exposed to ultraviolet (UV) light and other processes that causeenvironmental damage

1. Introduction
Bioremediation is a process that utilizes biological agents, such asmicroorganisms, to eliminate pollutants, making it a natural and efficientmethod for environmental cleanup. This approach is considered cost-effectiveas it transforms harmful pollutants into less toxic substances through microbialdegradation or detoxification. Currently, many developing nations areprioritizing industrial growth, urbanization, and the use of agrochemicals inagriculture, which has heightened the importance of bioremediation and drawnsignificant attention to this practice. As a sustainable solution, bioremediationoffers a way to restore healthier ecosystems by removing pollutants from theenvironment (Fig. 1).

Introduction

Bioremediation can be defined as elimination, attenuation or transformation of polluting or contaminating substances by the use of biological processes, to minimize the risk to human health and the environment. Bioremediation of contaminated soil and water uses microbes (e.g. bacteria and fungi) to convert pollutants to harmless or more environmentally acceptable products and minimizes the risk to human health in a natural or managed process.

9.1 The Cartagena Protocol on Biosafety: Objectives and Implementation
9.1.1 Introduction
The Cartagena Protocol on Biosafety (CPB) is a pivotal international legal instrument under the Convention on Biological Diversity (CBD), dedicated to regulating the transboundary movement, handling, and use of living modified organisms (LMOs) derived through modern biotechnology that may adversely affect biological diversity, human health, or the environment. Adopted in January 2000 in Cartagena, Colombia, and entering into force on September 11, 2003, the Protocol represents a global consensus on biosafety governance amidst the rapid expansion of genetic engineering technologies.
9.1.2 Objectives of the Cartagena Protocol
The principal objective of the Cartagena Protocol is:
“To contribute to ensuring an adequate level of protection in the field of the safe transfer, handling and use of living modified organisms resulting from modern biotechnology that may have adverse effects on the conservation and sustainable use of biological diversity, taking also into account risks to human health, and specifically focusing on transboundary movements.” (Article 1, CPB)
The core objectives can be summarised as
• Ensuring biosafety in the context of international trade of LMOs.
• Protecting biodiversity from potential risks posed by LMOs.
• Safeguarding human health against unintended exposure to transgenic organisms.

8.1 Steps in Biosafety Risk Assessment
8.1.1 Introduction
Biosafety Risk Assessment is a systematic, science-based process used to identify, evaluate, and mitigate risks associated with handling biological agents, genetically modified organisms (GMOs), and biotechnological products. It is central to ensuring human safety, environmental integrity, and regulatory compliance in both laboratory and field applications.
A well-conducted biosafety risk assessment enables institutions to establish appropriate biosafety levels, implement containment measures, and ensure sustainable and responsible use of biotechnological advancements.
8.1.2 Key Objectives of Biosafety Risk Assessment
• Prevent exposure of personnel to biohazards.
• Avoid environmental contamination and unintended gene flow.
• Determine containment requirements for biological agents.
• Guide institutional and national biosafety decision-making.
• Support regulatory frameworks and compliance obligations.

Abstract

Soil microorganisms play a major role in the biodissolution of nutrients bound in the soil silicate minerals. Plant nutrients like K, P, Ca, Mg, Fe, Zn, and Mn are bound in them. Silicon is considered as agronomically important as it confers rigidity and strength to crop plants and enhances yield. Biodissolution of these nutrients result in their availability to crops influencing the growth, biomass and yield. The microbial metabolites like organic acids, amino acids, inorganic acids, amines, ammonia, phenolic compounds, extracellular polysaccharides, siderophores, ligands of cations and cation exchange are involved in the biodissolution of K, Zn and Si. Frateuria aurantia, Bacillus mucilaginosus and B. edaphicus and Bacillus sp are recognized as potash mobilizers, silica and zinc solubilizers.

Abstract

Contamination of environment with heavy metals has become a serious global problem due to its threat to animal and human health. Removal of this non-biodegradable, hazardous heavy metals through bio-materials has received paramount interest and intense attention in the last few decades as bio-materials play crucial role in remediating heavy metal polluted environment through a process called “biosorption”. The term biosorption, therefore, has become a popular theme in recent years due to its relatively simple, inexpensive, eco-friendly with rapid mechanism of removing heavy metals from polluted environment. There have been burgeoning studies around the globe for alleviating the problem of heavy metal pollution by selecting and developing an effective bio-sorbent with minimum treatment process for field application. This review aims to provide vast information on types of biosorbent available, their exploitation for better efficiency and their economic attractiveness for the restoration of heavy metal polluted environment.

Biostimulants examines substances and microorganisms that enhance plant physiological processes, growth, development, and tolerance to environmental stress. The chapter distinguishes non-microbial and microbial biostimulants and discusses plant extracts, protein hydrolysates, animal-derived hydrolysates, humic substances, seaweed and macroalgal extracts, and secondary metabolites. Microbial examples include nitrogen-fixing and nutrient-solubilizing organisms, arbuscular mycorrhizal fungi, Trichoderma, Bacillus, Pseudomonas, microalgae, and cyanobacteria. Their implications for germination, metabolism, root development, crop quality, growth promotion, and abiotic-stress tolerance are described. The chapter provides a framework for understanding biostimulants as complementary tools within sustainable crop production and natural farming systems. in modern sustainable crop production for farmers worldwide under diverse agricultural production conditions.

Abstract

Anaerobic microorganisms occur in oxygen-free environments as they possess biomolecules and metabolic processes that have adapted to sustain them in the stressed environments. These biomolecules and metabolic processes are harnessed and employed in various biotechnological applications. Anaerobes are abundant in various ecological niches such as freshwater or marine sediments, subsurface aquifers, deep thermal vents etc. where, in the absence or limitation of oxygen, they utilise reduced ions such as nitrate, sulphate, ferric, carbonate and certain organic compounds like degrading organic matter, fatty acids or alcohols as electron acceptors during respiration/fermentation to produce more or fewer reduced products such as, alcohols, ammonia, organic acids, hydrogen and carbon dioxide as the oxidised product. Diversity of substrates that can be fermented and the wide range of products that can be formed make anaerobic processes useful in biotechnological applications. Their substrate versatility and flexibility, for example, they can utilize simple sugars as well as many of them can utilise complex renewable biomass make their biotechnological applications sustainable and environment friendly. Anaerobes have been employed in food, chemical and material industries for the production of food items, preservative, additives, chemical intermediates, solvents, catalysts and polymers. They find widespread usage in energy sector for generation of various kinds of biofuels that can mitigate our reliance on non-renewable sources for fuel production. An emerging aspect of anaerobic biotechnology is waste valorization that involves treatment of any kind of waste (industrial, municipal, agricultural, pharmaceutical), where organic fraction of the waste is converted into valuable products. Anaerobes play key role in maintaining the global cycles of carbon, nitrogen, and sulfur, and also facilitate bioremediation by breakdown of persistent compounds. Lately, role of anaerobes in medicine and health care sector is widely explored as they are found to produce several bacteriocins, antibiotics, immunomodulators, vitamins and anti-tumor compounds. Some of the anaerobes have also been employed as probiotics. In the present chapter, we discussed these and many more biotechnological applications of anaerobes in white, green, red and blue biotechnology.

1.    Introduction    496

2.    Strain Development Technologies    496

    2.1.    Screening of natural variants of microorganisms, 496
    2.2.    Genetic manipulation of microorganisms, 498
        2.2.1.    Random mutagenesis and selection, 498
        2.2.2.    Conjugation, 504
        2.2.3.    Transduction, 504
        2.2.4.    Transposition, 506
        2.2.5.      Hybridization, 507
        2.2.6.    Rare Mating, 509
        2.2.7.    Cytoduction, 510
        2.2.8.    Protoplast fusion, 511
        2.2.9.    Transformation, 511
        2.2.10.    Gene cloning and transformation (Recombinant DNA technology), 513

Introduction
Biotechnology represents a transformative force in modern agriculture, offering innovative solutions to enhance crop production, improve resistance to diseases, and address the global challenges of food security and climate change. Chapter 9 delves into the multifaceted role of biotechnology, particularly in the realm of genetic innovation, and how it is reshaping the agricultural landscape. The chapter begins by exploring the various applications of biotechnology in crop improvement. This includes the development of genetically modified organisms (GMOs) and transgenic crops, which have been engineered to exhibit desirable traits such as increased yield, pest resistance, and enhanced nutritional content. These advancements have significantly contributed to the global food supply, particularly in regions facing harsh climatic conditions or limited agricultural resources. A detailed examination of genomic tools and techniques follows, highlighting the sophisticated methodologies employed in biotechnology. Techniques such as CRISPR -Cas9 gene editing, marker-assisted selection, and genomic sequencing are discussed, showcasing their pivotal role in accelerating the development of improved crop varieties. These tools have enabled scientists to manipulate genetic material with unprecedented precision, paving the way for more efficient and targeted crop breeding programs. The chapter also addresses the role of biotechnology in crop protection, emphasizing its importance in developing crops that are resistant to pests, diseases, and environmental stresses. By enhancing the natural defense mechanisms of plants, biotechnology reduces the need for chemical pesticides, thereby promoting more sustainable agricultural practices and minimizing the environmental impact of farming. The importance of biotechnology and genetic engineering in ensuring food security is a central theme of this chapter. As the global population continues to grow, the demand for food is expected to increase dramatically. Biotechnology offers a viable solution to meet this demand by enabling the production of higheryielding and more resilient crops. This section also explores the ethical and regulatory considerations surrounding the use of GMOs and other biotechnological innovations in agriculture.

Southern Blotting
Introduction

Southern hybridization was named after Edward M. Southern who developed this procedure at Edinburgh University in the 1970s. To oversimplify, DNA molecules are transferred from an agarose gel onto a membrane. Southern blotting is designed to locate a particular sequence of DNA within a complex mixture. It is also called Southern blotting since the procedure for transfer of DNA from the gel to the nitrocellulose filter resembles blotting. In Southern hybridization, a sample of DNA containing fragments of different sizes is subjected to electrophoresis using either polyacrylamide or agarose gel.

Abstract

Higher plants can be exploited for the discovery of new bioactive products that could serve as lead compounds in pesticide development because of their novel modes-of-action. Different secondary metabolites of higher plants have been reported to exhibit efficacy as fungitoxicants as well as mycotoxin inhibitors. Pest controls using the botanicals are safer to the user and the environment because they break down into harmless compounds within hours or days in the presence of sunlight. The article presents different plant products found efficacious as antimicrobial and inhibitory to mycotoxin secretions by fungi. Their prospect as safe alternatives to synthetic antimicrobials in plant protection has been discussed.