eChapters

After planning research problem, the next step is to start real research work. Indeed, it is said that planning of a research work is only 20% of work done while conducting the experiments is 80% to be done and is more laborious, requiring extended time and patience of the scientist. While conducting your research work,it is essential to record each and every observation of your work. These observations will be of two types. One is the subjective observations which are difficult to quantify. For example, if you are recording clinical parameters of a disease,you may record pulse, temperature, respiration in arithmetic form but clinical symptoms are subjective and require a description of the patient rather than filling tabular sheet. In describing morphology of a parasite both parameters are needed ; you will describe morphology of the parasite and will also mention measurements of different parts of the parasite. Likewise, you may like to describe weather conditions or behavior of animals, vectors at certain stage of disease while you have to mention figures in many cases. Thus your observations are clearly divided in two categories – one is subjective observation and other is recording arithmetic values.

The very basics in one’s life is whether he has judged his interests or decided goals of his life or has taken life casually. The first is to know what you wish to achieve in your life or in the words of our former president and Bharat Ratna APJ Abdul Kalam (14) what are your dreams ? If you have dreams than only you can achieve them hence it’s essential to know what is your passion ?, passion is the key to success. In pursuing your passion, all things become secondary ; in fact, desired changes occur within you, unnoticed and automatic to achieve your goals. It may not be out of contest to mention that basically I belonged to a business family and being eldest son in the family, my father asked me to join family business, after doing my intermediate, with the liberty to join part time BA classes while carrying business activities. But business was not my passion hence I struggled to join to the academics and carried all my Veterinary education with distinctions - because of my interests. I am not sure what would have happened to my life have I joined least interested business line- perhaps a failed business man.

A close look on three disciplines of Parasitology reveal that these are the branches where a scholar initially select them as per his basic qualifications. But while pursuing research in Parasitology, the line of demarcation becomes blurred and it is in fact combination of a group of scientists,from varied disciplines, who are able to solve any one problem; at that point, it is absurd to divide the subject as medical or veterinary or basic parasitology. In fact, these will be your interests and experiences which will decide your line of research and will determine your achievements.

Livestock has been essential for Indian agriculture since prehistoric times. Livestock is crucial for tripling farmers’ revenue, for bovines—particularly cattle and buffaloes—play a crucial role in the economy. India has the
largest number of livestock owners, with a wide variety of livestock and poultry breeds that are essential to the socioeconomic advancement of rural communities. The production of consumables and maintaining the country’s food security are inextricably related to the livestock and agricultural sectors. Numerous illnesses have a detrimental effect on the health and productivity of livestock and are highly significant economically. Livestock illnesses have negative effects on producers, costing them money in upkeep, resources, and production.
Protozoal infections are a major contributor to the global burden of infectious diseases and have a profound effect on health, society, and the economy. Nearly one-sixth of the world’s population is at risk of dying from serious diseases like malaria (Plasmodium spp.), African sleeping sickness (Trypanosoma brucei), Chagas’ disease (Trypanosoma cruzi) and various forms of cutaneous and visceral leishmaniasis (Leishmania spp.). Although the burden of protozoal diseases primarily affects tropical and subtropical regions, environmental changes and ecological modifications brought about by both natural and manmade factors have had and are likely to continue to have a significant impact on the emergence and spread of these infections in high-income nations.

Introduction

Resistance to antibiotics in the bacterial world is a genetically determined phenomenon that was first observed within a few years after the introduction of antibiotics for therapeutic purposes. In Nature, ARGs are believed to have originated about three billion years ago as an intrinsic defense mechanism in antibiotic-producing bacteria (Perry et al., 2016; Iskandar et al., 2022), and subsequently, evolved by complex and quantum jump trajectories to mobilize, diversify and interconnect all ecosystems in the micro-biosphere. Bacterial genomes exhibit plasticity and continue to evolve under the selection pressure of antibiotics and other stressors (Patel, 2016). The ARGs in circulation at present are thus a mixture of ancient and modern counterparts. Recently, the interplay between bacterial metabolism and antibiotic resistance has also been revealed (Zampieri, 2021).

The use of antimicrobials in health management, livestock, and food industry is driving the emergence of resistant microbes at a rapid pace (Wall et al., 2016; Adegoke et al., 2016; Hu et al., 2017). As a result, AR pathogens have spread globally to cause loss of lives, production, productivity, and negatively impact economic growth. Recently, AR pathogens in a large study have been found directly responsible for millions of deaths, and the figures are likely to increase in the coming decades (Murray et al., 2022). The pace of the emergence of AR can however be slowed down by improving human interventions. The global and local AR combat plans implemented during the past few years have enhanced the mass awareness of the pandemic problem (WHO, 2015; FAO, 2016; NAP, GOI, 2017; WHO, 2022; WOAH, 2022). These Action Plans on AR containment encourage avoiding of indiscriminate use of antibiotics for the treatment of sick humans and animals, recommend the ban on the use of antibiotics as growth promoters in food animals, promote AR surveillance and monitoring, and support the ‘One-Health’ strategy and application of the biosecurity principles for containment of the resistant microbes.

The reclassification of the order Rickettsiales was predicated upon comprehensive genetic analyses of 16S rRNA, groESL, and surface protein genes. Consequently, organisms within this order were systematically assigned to one of two families: Anaplasmataceae and Rickettsiaceae. Within the Anaplasmataceae family, phylogenetic analyses consistently delineated four genetically distinct clades: (1) Anaplasma, (2) Ehrlichia, (3) Wolbachia, and (4) Neorickettsia. Despite the obligate intracellular nature of rickettsial organisms in both families, those classified under Rickettsiaceae proliferate freely within
the host cell cytoplasm, whereas members of the Anaplasmataceae family are confined exclusively to membrane-bound vacuoles within the cytoplasm of host cells.
The first documented cases of anaplasmosis trace back to the late 19th century in South Africa. However, it wasn’t until 1910 that the causative agent of the disease was conclusively identified morphologically as “marginal points” within red blood cells by Max Theiler, later named it Anaplasma marginale. A year later, he identified another species, A. centrale, which caused a milder form of the disease in cattle. During the 1930s, Sanborn, Stiles, and Moc demonstrated the mechanical transmission of A. marginale by horse flies and stable flies. In the late 1980s, the life cycle of A. marginale was fully elucidated in ticks belonging to the genus Dermacentor.

Organisms in the order Rickettsiales form a diverse group of small (0.3 to 0.5 x 0.8 to 2.0 pm), non-motile, pleomorphic Gram-negative bacteria which replicate only in host cells. They can be cultured in the yolk sac of embryonated eggs or in selected tissue culture cell lines. Because they stain poorly with aniline dyes, these organisms should be stained by Romanowsky methods such as Giemsa or Leishman. In addition to host-cell dependence and poor affinity for basic dyes, a requirement for an invertebrate vector distinguishes them from conventional bacteria and the Chlamydiales.
Epidemiology
Animal hosts and arthropod vectors are the reservoirs for most rickettsiae. A number of rickettsia1 organisms, including Ehrlichia canis, Anaplasma marginale and Haemobartonella felis produce latent infections. In arthropods, rickettsiae replicate in the epithelial cells of the gut before spreading to other organs, including the salivary glands and ovaries where further replication may occur.
Organisms are transmitted when the arthropod feeds on the animal host. Some organisms such as Rickettsia rickettsii are maintained in a tick population by transovarial transmission. Trans-stadial but not transovarial transmission of E. canis and E. phagocytophila occurs in ticks. With the exception of Coxiella burnetii, which produces endospore-like forms and can remain viable in dust for up to 50 days, most rickettsiae are labile outside host cells. Aerosol transmission of C.burnetii commonly occurs in domestic animals and humans. In addition, a silent cycle involving ticks and small wild mammals may constitute a possible source of infection for some domestic species.
Pathogenesis and Pathogenicity
Many Rickettsia species including the causal agents of typhus (R. prowuzekii), murine typhus (R. typhi) and scrub typhus (R. tsutsugamushi) are primarily human pathogens. Rocky Mountain spotted fever, caused by Rickettsia rickettsii, which is a common rickettsia1 disease of humans, also affects dogs.

Introduction
Antimicrobial resistance (AMR) is an escalating global concern, rendering many once-effective antimicrobials useless and threatening modern medicine’s foundations [1]. RNA-based therapies offer precision, programmability, and modularity—tailoring interventions directly to microbial genetic machinery [2, 3].
Main RNA-based modalities include
• Antisense Oligonucleotides (ASOs): Short synthetic single-stranded RNAs that bind complementary mRNA in bacteria to inhibit translation or induce RNase H–mediated degradation [15].
• RNA Interference (RNAi): Though classically applied to eukaryotes, siRNAs or crRNAs delivered via nanoparticles (also termed “nanoantibiotics”) can silence microbial resistance or virulence genes while the carrier materials physically damage microbial membranes— enhancing lethality [3, 4,5].
• CRISPR-Cas Systems: Adapted as antimicrobials, CRISPR-Cas tools can precisely target and cleave resistance genes in pathogens via conjugative plasmids or phages [6].
• Ribozymes / External Guide Sequences (EGSs): Engineered noncoding RNAs that harness RNase P or ribozyme activity to degrade specific bacterial mRNAs [16].
Key strengths
• Target specificity—can silence essential microbial or resistance genes while sparing beneficial microbiota.
• Rapid design and adaptability—once delivery systems are in place, targeting new genes requires sequence change alone.
• Synergy potential—combining gene silencing with antibiotics or physical antimicrobial actions increases efficacy.

Robotics is transforming livestock production by automating repetitive, labour-intensive and precision-dependent operations. This chapter examines automated milking, robotic feeding, manure removal, health monitoring, drones and autonomous herd-management systems across dairy, poultry, swine and sheep production. Integration with AI, IoT and sensors creates connected environments capable of continuous monitoring and data-driven action. Benefits include higher productivity, lower labour requirements, timely animal care and more efficient resource use. Challenges include substantial investment, technical complexity, maintenance and farmer acceptance. Emerging developments such as autonomous farms, swarm robotics, solar-powered equipment and climate-smart robotic systems may further strengthen animal welfare, environmental sustainability and global food security.

Jay et al., (1995) and Joerger et al., (2003) have written the role of bacteriocins as an alternate to antibiotics. Bacteria produce bacteriocins which are proteinaceous in nature or peptidic toxins. These inhibit the growth of similar or closely related bacterial strain(s). There are different types of bacteriocins which are classified based on the producing strain, common resistance mechanisms and mechanism of killing. Ahmad et al., (2013) have isolated antifungal compound for food preservation. Martin et al., (2018) and Batish et al., (1997) found antibacterial compounds from fungus. Mahboubi et al., (2019) studied the potential of Zataria in IBS. There are bacteriocins from gram-positive bacteria known as the colicins, the microcins and the bacteriocins from Archaea (Cascales et al., 2007). The bacteriocins from E. coli are called colicins (formerly called ‘colicines,’ meaning ‘coli killers’). These have been studied widely. Earlier called colicin V and is now known as microcin V. It is very small and is produced and secreted in a different manner than the classic colicins. Adrienne et al., (2016) have studied the colicin resistance.

Introduction
Antimicrobial resistance (AMR) has emerged as one of the most alarming public health challenges of the 21st century, threatening the effectiveness of life-saving drugs and the future of modern medicine. This crisis is not limited to healthcare settings; it spans communities, agriculture, and the environment, impacting every facet of society. India, with its large population, high burden of infectious diseases, unregulated access to antibiotics, and considerable pharmaceutical manufacturing industry, stands at the forefront of the global struggle against AMR. The rise of resistant microbes stems from a complex interplay of factors: overuse and misuse of antibiotics in humans and animals, poor infection prevention practices, lack of sanitation, and environmental contamination from antibiotic waste. As a result, common infections are becoming harder to treat, surgical procedures riskier, and healthcare costs are escalating, all leading to increased morbidity and mortality. Tackling AMR requires robust intersectoral collaboration—no single stakeholder can succeed alone. Non-governmental organizations (NGOs), industries, and Corporate Social Responsibility (CSR) initiatives form a critical triad in India’s fight against AMR. NGOs drive grassroots education, advocacy, surveillance, and policy change, often bridging gaps where governmental capacity is limited. The pharmaceutical and healthcare industries, responsible for antibiotic manufacturing, distribution, stewardship, and infection control, play a pivotal role in curbing environmental emissions, supporting responsible use, and promoting the discovery of new treatments. Meanwhile, CSR channels the resources and expertise of corporations towards sustainable, communityfacing projects that promote awareness, improve hygiene, foster stewardship, and drive systemic change.

There are number of internal and external factors that affect the production performance of animals, among which parasites and parasitism are major of concern constraints to animal’s productivity especially in tropical countries. Feed intake and nutrient requirements have been recognized for long time as an important parameter for determining the efficiency of animal production. Most of the parasitic infection in domestic animals is caused by gastrointestinal nematode, which may result in depression in appetite, impairment in GI functions, alteration in protein, energy and mineral metabolism and changes in water balance. Nutrition can affect the ability of the host to cope with the consequences of parasitism and to contain and eventually to overcome parasitism. Since it is proposed that the host gives priority to the reversal of the pathophysiological consequences of parasitism over other body functions, it is to be expected that improved nutrition will always lead to improved resilience. On the other hand, it is proposed that the function of growth, pregnancy and lactation are prioritized over the expression of immunity. Improved nutrition may affect the degree of expression of immunity during these phases and can thus influence the resilience and resistance of host to parasite infection.

In this century the usage of plastics had become very common. However, it is important to know as to what kinds of plastics are used for food handling and storage and the health hazards caused by using it. It has been observed that plastic packaging plays a significant role for the enhanced shelf life and in the ease of storage and cooking for many foods. Some kinds of plastic materials which are widely used for handling and storage of food and water include polyethylene terephthalate (PET). It is used in soft drinks, water, sport drinks, ketchups, salad dressing bottles, peanut butter, pickle, jelly and jam jars. This plastic is strong, heat resistant, resistant to gases and acidic foods. It can be transparent or opaque. It does not leach any chemicals that are suspected of causing cancer or disrupting hormones and can also be recycled.

Usage of High density polyethylene (HDPE) has been found to be common in milk, water, juice bottles, yogurt, margarine tubs, grocery, trash and retail bags. High-density polyethylene is stiff and strong but is not heat stable (it melts at a low temperature). It does not leach any chemicals that have a role in causing cancer or disrupting the hormones. HDPE can be recycled. Low-density polyethylene (LDPE) is used into making films of various types, some bread and frozen food bags and also in the making of squeezable bottles. Low-density polyethylene is relatively transparent. Many of the films are not heat stable either and may melt the food in case it is touching it. However, polypropylene (PP) is more heat resistant, harder, denser and more transparent than polyethylene and is therefore used for heat-resistant microwavable packaging and sauce or salad dressing bottles. Polycarbonate is clear and heat resistant. It is also durable and often used to make refillable water bottles and sterilisable baby bottles, microwave ovenware, eating utensils, plastic coating for metal cans. Tiny amounts of bisphenol A are formed when polycarbonate bottles are washed with harsh detergents or bleach (sodium hypochlorite). At high levels of exposure, bisphenol A is potentially hazardous as it mimics the female hormone estrogen. In addition, polystyrene (PS) and polyvinyl chloride (PVC) are also used during food material transportation and handling in supermarkets. Modern food safe plastic bags are plasticizer-free which do not leach harmful chemicals into the food during cooking.

Due to the globalization of the food supply there has been a need to strengthen food safety systems in and between the countries. In this regard the World Health Organization (WHO) has promoted efforts in improving the food safety from farm to plate which started on World Health Day, 7 April 2015. The World Health Day 2015 slogan was “From farm to plate, make food safe”. The informal food production and marketing system is still strong in most countries of the South-East Asia Region, for the enforcement of food safety regulations. Traditionally, the preferences have been for hot and well-cooked food. These habits are partially responsible for preventing foodborne infections. In many countries street food has become very popular as it is cheap. Hygienic conditions are improving, provided that potable water supply and clean facilities are ensured by municipal authorities. Many waterborne and diarrhoeal diseases could be prevented with the introduction of bottled drinking water. The awareness and education on food safety by the consumers has strengthened in the enforcement of food standards. This has improved the hygienic practices and has helped in the prevention of foodborne illnesses. The “WHO Five keys to safer food” serves as the basis for educational programmes to train food handlers and educate the consumers. These help in preventing the foodborne illnesses. The important points are to keep food surfaces clean, washing of all utensils, plates and cutlery, keeping the raw food separate from the cooked food. Cooking food thoroughly to the appropriate temperature, keeping food at safe temperatures, both for serving and storage and using safe water and raw materials.

For a researcher, laboratory is another home where he/she spends much time every day. As we take care of cleanliness, safety, and design of our home, we must take care on these aspects of laboratory too. It is the responsibility of the laboratory personnel to ensure cleanliness/safety of the laboratory. Since every laboratory chemical and equipment have the potential to cause harms if not used with appropriate precautions. Hence, as a laboratory user, it is our responsibility to follow the basic safety/biosafety rules and regulations set up by the Principal Investigator of the laboratory, Institutional Biosafety Committee, as well as by other regulatory authorities, if any. We must be aware of the general/personal protective measures, cleanliness and laboratory waste disposal prior to joining a research laboratory. In this chapter we will briefly discuss about personal safety, chemical safety, cleanliness, waste disposal, etc.

A. Personal Safety

1. Personal Protective Equipment (PPE)

Personal protective equipment (PPE) is special arrangement to protect the worker from specific hazards because of using a hazardous substance. PPE does not reduce or eliminate the hazard, but protects the wearer from exposure to hazardous material. PPE should not be reused or brought outside the laboratory as these might increase the risk of spread hazardous material or contamination. Used PPE must be kept inside the laboratory in a designated waste storage area adjacent to the work area.

Josephson (1977) studied the effects of food irradiation and sterilization. In radiation sterilization of food (radappertization) the food is exposed in sealed containers to ionizing radiation at absorbed doses which are high enough (25-70 kGy) to kill all organisms of food spoilage and public health significance. Radappertization is analogous to thermal canning for enhancing the shelf stability. In dry products the radappertization process requires the food to be heated to an internal temperature of 70-80°C (bacon to 53°C) to inactivate autolytic enzymes which catalyze spoilage during storage without refrigeration. In order to reduce off-flavors and odors, undesirable color changes, and textural and nutritional losses from exposure to the high doses of irradiation the foods are vacuum sealed and irradiated frozen. Radappertozed foods have the characteristic of fresh foods prepared for eating. Irradiation can be a good substitute for some chemical food additives such as ethylene oxide and nitrites which are toxic, carcinogenic, mutagenic or teratogenic either wholly or in part. Till now, there is no evidence of any adverse effects of radappertization on the foods. Extensive research has taken place towards the space food. During the planning of Apollo missions, Space Food NASA Technical Reports Server (Space Food NASA Technical Reports Server, NTRS ,1994 ) conducted trials including freeze drying.