eChapters

Introduction

Test of pollen viability (extent of pollen viability) is important in pollination experiments to ensure fertility/seed setting. It is also important in monitoring state of pollens during storage in ecological/ taxonomic studies as well as during research on pollen biochemistry, genetics, stigma interactions, and incompatibility studies. Pollen viability refers to the ability of pollen to accomplish post-pollination events to achieve fertilization. Pollen viability test has gained importance particularly in plant breeding and hybrid seed production experiments/ practices. In this chapter, you will be acquainted with the basics of pollen viability test. However, for more professional knowledge on the topic readers are suggested to refer specialized books.

Pollen Viability Test

It is important to know the extent of viability of pollens used in pollination experiment, in finding fertility of a plant of interest, to monitor state of pollen on storage, in ecological/taxonomic studies, as well as in research on biochemistry, genetics, stigma interaction, incompatibility assay, etc. of pollen. Several pollen viability tests have been developed time-to-time. It is recommended to initially use different tests for a given pollen type to find out the most appropriate method that provides reliable result on pollen viability. It is important to note that every procedure requires standardization of protocol according to the sample used. Following different tests are used for assessing pollen viability.

1. In-vitro Pollen Germination Test

It is a simple, speedy, and quantitative test for assessing pollen viability. In this method, a collection/sample of pollen is germinated in vitro and observed under microscope. The percentage of germinating pollens (producing pollen tube) after appropriate treatment and a given period of time is determined. The percentage provides an index of viability of the pollen sample.

Globally, antimicrobial resistance (AMR) is an important public health and development threat (WHO, 2020). Being one of the top 10 global public health threats, the ever-increasing rates in the occurrence of drug-resistant bacterial infections have become a grave public health concern for the governments (Laxminarayan et al., 2013). Moreover, such infections would be difficult to treat and therefore can lead to worse clinical outcomes (Laxminarayan et al., 2013). Worldwide, each year about 700,000 people die from antimicrobial resistant infections and the mortality figures are projected to reach 10 million/ year by 2050 (O’Neill, 2016). The crude infectious disease mortality rate in India is more than 400 per 100,000 persons and at least 23,000 deaths in adults and 58,000 neonatal deaths per year are caused by AMR bacteria (Laxminarayan and Chaudhury, 2016). A 2017 estimate puts the cost of developing an antibiotic at around US$1.5 billion (Towse, et al. 2017). Meanwhile, industry analysts estimate that the average revenue generated from an antibiotic’s sale is roughly $46 million per year. “That’s tiny and nowhere near the.

Microbial Resistance Mechanism

Huge use of antibiotics in nosocomical environment has lead to the occurrence of multidrug resistance mechanism by microorganism against particular antibiotics (Munita et al., 2016; Zhao et al., 2020). Main operation includes chemical modification, efflux pump through passivation, modification of drug targeting gene and systemic elimination of antibiotics as many of them can form biofilm (Rajput et al., 2018) which can show extreme resistance. Chemical and enzymatic mechanism can include inactivation, activity loss, degradation and derivatizaton of antibiotics chemical group (Liu et al., 2019). Best idea may include production of enzymes or they can also expel antibiotic to efflux pump. Antibiotics drug efflux resistance can be accomplished by drug efflux pump playing an important role in microbial resistance. A different type of efflux pump system has been identified that include lipophilic and hydrophilic efflux system targeting the drug at different chemical properties (Wasaznik et al., 2009; Kumar et al., 2012). Another strategy adopting in resistance mechanism is modification of drug targeting gene. This mechanism makes the changes in drug to lose its target through drug targeting gene modification. Another important resistance mechanism is effectiveness of antibiotics by interfering with the target site. Therefore, bacteria has evolved different mechanism that include target site protection. A bactericidal role played by beta lactum antibiotics in inhibition of mucopeptide synthase and bacterial penicillin binding protein (PBP) in cell wall transpeptidation reaction that prevents the formation of complete call wall and hence dies (Sauvage et al., 2016). Bacteria also developed its own strategies to cope up with the environmental stress that can also include human body. In course of gaining the advantage bacteria needs to compete for nutrients and to keep away from the molecules produced by other competing organism (Zhao et al., 2019). Recently researchers have found the importance of biofilm resistance in bacteria. Bacterial biofilm resistance produce by complex and systematic drug resistance mechanism.

Cell culture medium : To grow cells in in vitro, environment should be as close as possible to in vivo condition. But a number of factors are important to grow cells in vitro namely substrate, temperature and medium. There is a number of ready made medium available commercially and made the cell culture work easier. Any successful medium consists of isotonic buffer solution containing inorganic salts, amino acids, vitamins, a energy source and various supplements. Usually serum or serum replacement is used as supplement.

Introduction

Antibiotic-resistant bacteria are a major cause of health care-associated infections around the world, and resistance has also emerged in infections in the wider community. Infections caused by multi resistant organisms significantly increase morbidity, mortality, and health care costs. Molecular analyses have revealed that widespread multi resistance has commonly been achieved by the acquisition of preexisting determinants followed by amplification in response to selection. The capture, accumulation, and dissemination of resistance genes are largely due to the actions of mobile genetic elements (MGE), a term used to refer to elements that promote intracellular DNA mobility (e.g., from the chromosome to a plasmid or between plasmids) as well as those that enable intercellular DNA mobility.

Insertion sequences (IS) and transposons (Tn) are discrete DNA segments that areable to move themselves (and associated resistance genes) almost randomly to newlocations in the same or different DNA molecules within a single cell. Other elements, such as integrons (In), use site-specific recombination to move resistance genes between defined sites. As these types of MGE are often present in multiple copies indifferent locations in a genome, they can also facilitate homologous recombination(exchange of sequences between identical or related segments). Intercellular mechanisms of genetic exchange include conjugation/mobilization (mediated by plasmidsand integrative conjugative elements [ICE]), transduction (mediated by bacteriophages), and transformation (uptake of extracellular DNA). Interactions between thevarious types of MGE underpin the rapid evolution of diverse multi resistant pathogens in the face of antimicrobial chemotherapy. Primarily the most important and/ or topical elements in Gram-negative and Gram-positive bacterial species of particular concern clinically, namely, Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. (the so-called ESKAPE group) (Rice LB., 2008) as well as Escherichia coli (giving the ESKAPEE group) (Llaca-Díaz J.M. et al.,2012); nonetheless, many of the MGE types described are of course broadly relevant to many other bacterial taxa.

Performing animal means any animal which is used at, or for the purpose of any entertainment to which the public are admitted through sale of tickets. Thousands of wild animals are used worldwide to perform demeaning and unnatural tricks to entertain the public (circuses, side-shows, within zoos, and in advertising, film and television). 
There are number of welfare issues related to performing animals which involves animal fighting, shows, (among domestic animals) or entertaining wild animals.

Long used as beasts of burden, animals in India are still regarded today as little more than commodities rather than sentient beings who feel pain, stress, exhaustion and depression. Low literacy rates and the tendency to follow traditional practices and cultural traditions have left India surely lacking in the area of animal welfare. India holds the majority of the world’s working animal population. Animals that are forced to work in India include bullocks, horses, ponies, mules, donkeys, camels, elephants and dogs. Despite a proud heritage and a culture rooted in tolerance and compassion, animal welfare is largely ignored by Indian society, leading to extreme animal suffering. Our villages and to a great extent, the urban population still allow working animals to be mercilessly exploited in fields, on the streets, in brick kilns, on battlefields, on playgrounds and in timber areas – all in the name of agriculture, entertainment, security and any number of other “justified” pursuits.

Companion animals are all those species of animals which are kept by man for companionship and which are often referred to as pets. It is quite common to see community dogs or cats living in residential colonies. This is because due to very fast urbanization all the open spaces are getting used up for construction of residential complexes, and the animals like dogs and cats living on these open areas are getting displaced. The residents that occupy these residential buildings find these animals to be a nuisance and want them to evicted.

Pet and companion animal can be discussed under street dog/stray dogs and owner dogs.

1.1     Prokaryotic 
    
1.2     Eukaryotic
    
1.3     Cell orgenellie

4.1 DNA: The word means
    
4.2 Double Helical Structure: The Watson and Crick Model
    
4.3 Family of DNA 
    
4.4 The Chromosomes

Introduction
Antimicrobial resistance (AMR) poses an escalating threat to global health, eroding the efficacy of antibiotics and complicating the management of bacterial infections. The proliferation of resistant pathogens, such as methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum betalactamase (ESBL)-producing Enterobacteriaceae, and multidrug-resistant Mycobacterium tuberculosis, has driven significant increases in morbidity, mortality, and healthcare costs. The ability to detect AMR accurately and promptly is paramount for guiding clinicians in selecting effective therapies, implementing targeted infection control measures, and informing public health strategies to curb the spread of resistant organisms. Conventional methods, rooted in phenotypic assessments, have served as the cornerstone of AMR detection in clinical microbiology laboratories for decades. These techniques evaluate bacterial growth under controlled exposure to antibiotics, offering direct evidence of resistance or susceptibility that molecular approaches, which focus on genetic markers, may miss due to silent mutations, complex regulatory mechanisms, or novel resistance pathways. This chapter provides an exhaustive examination of conventional AMR detection methods, encompassing disk diffusion, broth and agar dilution, gradient diffusion, and automated susceptibility testing systems. These approaches, refined through decades of clinical and research application, remain indispensable for their reliability, cost-effectiveness, and adaptability across diverse settings, from well-resourced tertiary hospitals to laboratories in low-income regions. By exploring their principles, procedures, applications, and limitations, we aim to underscore their enduring significance amidst the rise of rapid molecular diagnostics. The discussion also addresses specialized tests for detecting specific resistance mechanisms, such as beta-lactamase production, inducible clindamycin resistance, and carbapenem’s activity, which are critical for tailoring antimicrobial therapy. Through a critical lens, this chapter evaluates the strengths and challenges of these methods, offering insights into their role in addressing the global AMR crisis, their integration with emerging technologies, and their continued relevance in both routine diagnostics and epidemiological surveillance. The value of conventional methods lies in their ability to translate laboratory findings into actionable clinical decisions. Unlike genotypic methods, which detect resistance genes but may not predict phenotypic behaviour, these techniques assess how bacteria respond to antibiotics under conditions mimicking in vivo environments, aligning closely with therapeutic outcomes. As resistance patterns grow increasingly complex, driven by mechanisms like carbapenem’s and efflux pumps, conventional methods provide a practical foundation for diagnostics, particularly in resource-limited settings where advanced molecular tools are often inaccessible. Their role in global AMR surveillance, such as through the World Health Organization’s Global Antimicrobial Resistance Surveillance System (GLASS), further underscores their importance in tracking resistance trends and informing policy.

Good livestock welfare practices demands thorough understanding of the basic nature of the animal. The science of understanding the details of animal behaviour is known as Ethology.  Knowledge about the behaviour of the species being reared proved as an important tool for implementing animal welfare in practical husbandry practices. Now a day’s global community is also advocating for the better livestock welfare standards. Exploiting animal’s natural behaviour could certainly prevent unnecessary sufferings related to handling and restraining during routine farm activities. On the one hand this ensures physical and mental welfare of livestock, while on the other hand improvement in the production is guaranteed. Without involving heavy investment behavioural knowledge based management interventions can bring paradigm shift in welfare status of farm. Therefore, behavioural studies are of great importance in increasing our understanding and appreciation of animals.

India is known for its rich wildlife heritage. In fact, the country is considered one of the best among available mega-biodiversity areas in the world. The landscape and agro-climatic conditions of India make it a unique habitat for wild animals. The government of India has made attempts to conserve wildlife through different projects like Corbett tiger, Elephant projects etc. The wild life management in India is at present ‘preservation-oriented’. This is bound to remain so for a long time because of the significantly depleted status of our wild life population. Indian government in pursuit of conserving the heritage of wilderness has made committed efforts with several wildlife and Forest Acts to ensure protection by declaring the areas as protected under the spectrum of national park and wildlife sanctuaries. The eco-tourism and conservation awareness has created environmental consciousness among the people.

9.1 Methylases
    
9.2 Polymerases
    
9.3 Nucleases
    
9.4 Ligases
    
9.5 Phosphatase
    
9.6 Topoisomerase
    
9.7 Restriction enzymes

Introduction
Antibiotic-resistant E. coli develops as a result of the heavy use of antibiotics in the production of food animals, which is then found in the meat from those animals. There have been reports of a number of antimicrobial resistant E. coli infections linked to the ingestion of tainted milk and chevon. As a result, the existence of antibiotic resistant E. coli is a significant and expanding global public health risk. Escherichia coli of animal origin has also been seen to colonise humans, and because it is resistant to many conventional antibiotics, it can cause serious illnesses for which there are few effective treatment choices. Due to its ubiquitous presence in the environment, E. coli has traditionally been recognised as the main vehicle for the transmission of various antibiotic resistance genes and vectors.Extended-spectrum beta-lactamases (ESBLs), which can make the bacteria resistant to some frequently used antimicrobial treatments, are tiny proteins (enzymes) that some strains of E. coli have been shown to manufacture under favourable conditions. A major issue around the world is the emergence of different E. coli strains that produce ESBL in foods that come from animals that are raised for food. The ESBL enzymes TEM, SHV, and CTX-M from Group A have been widely reported to be generated by E. coli and have been shown to hydrolyze ampicillin, carbenicillin, oxacillin, and an extended spectrum of cephalosporins, including ceftazidime and cefotaxime.
Extended beta spectrum lactamase producing E. coli
The occurrence of antimicrobial resistance and the presence of corresponding resistance genes in the environment is an ancient phenomenon. This is largely due to the fact that many of today’s antimicrobial agents are derived from natural precursors produced by soil bacteria such as Streptomyces. These natural compounds likely evolved to mediate microbial competition for

Introduction
Antimicrobial resistance (AMR) is a global concern that poses a significant threat to contemporary healthcare systems, potentially hindering the management and treatment of various diseases. Microorganisms can act as reservoirs for AMR across various ecological niches; thus, a “one health” proposal is essential for coordinating a multisectoral strategy aimed at investigating and tackling this alarming issue. This method seems to be an effective strategy for addressing and alleviating the challenges posed by AMR; however, it demands the collaborative efforts and assets that are consistently and efficiently executed by health practitioners. Horses have been pivotal in human history; they have served in wars, provided transportation, and even aided in mining operations. In contemporary society, horses are significant as sport animals and in therapeutic settings involving animals. Given these close interactions between humans and equines, it becomes essential to effectively identify infectious diseases and antimicrobial resistance (AMR) impacting both species, particularly in instances of highly contagious diseases. Resistant bacteria represent a significant threat to the health of both equines and humans. Horses may obtain resistant bacteria through several avenues, such as interaction with other animals, exposure to contaminated materials, or to antibiotic uses. The responsible utilization of antibiotics and the implementation of antimicrobial stewardship programs are essential in reducing the transmission of antimicrobial resistance (AMR) within equine populations. A variety of zoonotic pathogens that exhibit resistance to antibiotics has been identified in equines. This includes Extended-Spectrum Beta-Lactamases (ESBL) which produces E. coli, Methicillin-Resistant Staphylococcus Aureus (MRSA), and Multidrug-Resistant (MDR) Salmonella. Horses can develop AMR through various channels like:

Animal’s response  to external stimuli is a behavior. Domestic animal behaviour and performance are correlated. Man’s intervention has led to huge modifications and modulations in animal behaviour patterns over ages but certain characters and normal behaviours of animals are needed to be expressed, otherwise may lead to abnormalities or performance loss. Animal freedoms also ensure certain basic freedoms to domesticated animals including freedom to express normal behaviour. With the developing concern over animal welfare among consumers, growers and animal activists, the need to ensure animal freedoms is call of the hour. Measuring animal welfare is a complex process which requires multidisciplinary approaches. There are four approaches to determine animal welfare viz, productivity, animal health and disease, physiology and behavior which are used individually or in combination.

Early and accurate diagnosis of disease is of prime importance for both the successful treatment of the patient and in limiting the spread of disease in the community. It is in this context that immunodiagnostic methods are now being used extensively since they can be extremely sensitive and highly specific. Among the most successful methods are those utilizing labelled antibodies or antigens. Examples of these are immunofluorescence in which antibodies labelled with fluorescent dyes are utilized and radioimmunoassay in which antigens or antibodies are labeled with isotopes. Both of these procedures have proved to be of great value for particular purposes.

Introduction 
The discovery and introduction of antimicrobial agents to clinical medicine was one of the greatest medical triumphs of the 20th century that revolutionized the treatment of bacterial infections. However, the gradual emergence of populations of antimicrobial resistant pathogenic bacteria, resulting from use, misuse and abuse of antimicrobials has today become a major global health concern. Antibacterial drugs have revolutionized our ability to control bacterial disease, and their clinical availability has led to dramatic decreases in morbidity and mortality. (Walsh and Wright, 2005). As such, these therapeutics underpin modern medicine. Despite the integral role of antibiotics in sustaining our modern lifestyle, they are undervalued in both cost and significance by society. Over the past century, their use has provided strong selective pressure on microorganisms, leading to preferential survival and spread of those harbouring antibiotic resistance mechanisms. Multidrug resistance is now commonplace amongst bacterial pathogens with antibiotic resistance now affecting all antibiotic classes. (Coates et al, 2011). This is particularly worrisome in the case of Gram-negative bacteria (e.g. Pseudomonas aeruginosa and Acinetobacter baumannii) for which treatment options are already limited. The ‘broken’ economics of antibacterial research and development is often quoted as the main reason for the lack of new therapies but the truth is that it is hard to discover new antibacterial drugs, and the science is not sufficiently advanced to allow discovery of efficient and effective drugs. This has led to fears of a ‘post-antibiotic era’ as it has been estimated that 5–20 novel antibacterial drugs need to enter clinical development in order to effectively contend with the current resistance problem. However, given the attrition rate within the existing drug discovery model, at least 200 discovery programmes would be needed in order to achieve this outcome. Hence, new approaches to antibiotic discovery are needed. The antibiotic pipeline The antibiotic pipeline is not what it once was.4 Pharmaceutical companies were once the main provider of novel antibiotic molecules but they largely withdrew in the late 1990s because of the lack of success and low financial returns in bringing new antibacterial drugs to the market.5 The environment of discovering and developing new antibiotics was quite different during the so-called ‘golden era’ of drug discovery. Antibiotics worked remarkably well because resistance was low and physicians had access to a variety of efficacious antibiotics. Antibiotic research and development programmes were inclined to focus on improved pharmacology to achieve less frequent dosing e.g. once a day, rather than innovative new antibiotics. Natural product screening strategies tended to result in rediscovery of compounds rather than finding new ones. There was also no need to consider those natural products with undesirable properties such as toxicity. Today, only a few large companies such as GlaxoSmithKline, Novartis, Merck and Roche are still actively engaged in antibiotic R&D, with many of the original antibiotic provide.The natural world remains the largest source of novel drug scaffolds making this a viable option in the search for new antibiotic compounds. Advances in bacterial culture techniques, molecular biology and metagenomics will make natural product drug discovery easier and more costeffective, obviating these limiting factors. Screening procedures must include whole-bacterial cell assays, addressing the issue of bacterial permeability and efflux early in the discovery process (Shore and Coukell, 2016). Additionally, the generation of training schemes by, and with, pharmaceutical companies that cover all aspects of the pipeline and include natural product drug discovery, are essential and will ensure that expertise is passed on to future researchers. Investment should also be made into the study of previously characterized lead compounds that did not reach the clinic, so called old leads. The reasons that led to these compounds being dropped from further development vary, ranging from financial issues to trial design, dosing problems and toxicity. It may be that there is now sufficiently improved technology and expertise to develop these as safe and efficacious antibacterials. The revival of interest in old leads could also provide an additional source of novel antimicrobials. A freely accessible database of antibiotics that were never developed has recently been launched, Antibiotic with the aim to reduce unnecessary duplication of Pharmaceutical agents such as nanospheres, liposomes, and immunoglobulins for example have been developed which can alter the pharmacodynamics of a compound, act as carriers for sustained or controlled drug release, and for immunization therapy.

Introduction
Antimicrobial resistance (AMR) is a mounting global crisis that threatens public health, food security, and development. The rapid spread of resistant pathogens has rendered many conventional antibiotics ineffective, leading to longer illnesses, increased mortality, and escalating healthcare costs(1)(2). The World Health Organization characterizes AMR as a top-ten global health threat, driven by overuse, misuse, and inadequate dosing of antibiotics. It already kills 700,000 people a year and is expected to kill 10 million people a year by 2050 if nothing is done about it(3). After penicillin was introduced to the market in 1941, the first signs of resistance appeared in staphylococci, streptococci, and gonococci. By 1942, Staphylococcus aureus, which was penicillin-resistant, emerged(4). Drug resistance must be effectively addressed for a few reasons, including public health, economic considerations, and the long-term viability of medicine. Maintaining the effectiveness of existing treatments for infectious diseases is an important objective(5). NDDS technologies have emerged as powerful tools to enhance the delivery, bioavailability, and targeting of antimicrobial agents, potentially reviving older antibiotics, and maximizing the efficacy of novel compounds(6). Parallelly, AI has found impactful applications in biomedical data analysis, drug discovery, and the rational design of therapies by processing complex datasets, predicting resistance patterns, and guiding the creation of new molecules. The integration of AI with NDDS marks a paradigm shift to optimizes delivery systems, guides individualized treatment plans, and advances the field towards precision medicine for infectious diseases (7). This chapter surveys the convergence of these technologies, exploring how AIenhanced NDDS platforms are reshaping the response to AMR. Additionally, it examines diverse AI models and algorithms employed in the prediction, optimisation, and management of drug resistance, enhanced by real-time case studies for thorough comprehension(fig1.).