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The mycoplasmas are microorganisms in the class Mollicutes. Of the nine genera in this class, five contain species of veterinary interest. The genus Mycoplasma, in which there are about 100 species, contains most of the animal pathogens. The first mycoplasma identified in 1890 was Mycoplasma mycoides subspecies mycoides, the cause of contagious bovine pleuropneumonia. Similar types of mycoplasmas which were subsequently identified were called pleuropneumonia- like organisms (PPLO).
Mycoplasmas, the smallest prokaryotic cells capable of self-replication, are pleomorphic organisms ranging from spherical (0.3 to 0.9 pm in diameter) to filamentous (up to 1.0 μm long). Because they cannot synthesize peptidoglycan or its precursors, they do not possess rigid cell walls but have flexible, triplelayged outer membranes.
Their flexibility allows them to pass through bacterial membrane filters of pore size 0.22 to 0.45 μm. Mycoplasmas are susceptible to desiccation, heat, detergents and disinfectants. However, they are resistant to antibiotics such as penicillin which interfere with the synthesis of bacterial cell walls. Based on 5s rRNA sequence analyses, the mycoplasmas have been shown to be linked phylogenetically to Gram-positive bacteria such as Clostridium species which have low guanine-cytosine content in their DNA. They require enriched media for growth, characteristically forming umbonate micro-colonies
when illuminated obliquely and microcolonies with a ‘fried egg’ dppearance in transmitted light. The dense central zone is due to extension of the microcolony into the agar. Mycoplasmas, which have relatively small genomes (approximately 800 genes) are fastidious in their growth requirements.
Most mycoplasmas are facultative anaerobes and some grow optimally in an atmosphere of 5 to 10% CO2. Nonpathogenic anaerobic mycoplasmas are found in the rumens of sheep and cattle. The genera Mycoplasma and Ureaplasma contain animal pathogens.

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.).

Our nature is the greatest or only laboratory where a large number of events are occurring at every fraction of a second - there are so many events, creatures, miseries taking place in the nature. Rightly it is considered an unparallel and biggest laboratory, man has ever witnessed. However, understanding it’s any event requires deep devotion and understanding,then only success will come. Our Philosopher and Poet Kabirdas has understood these facts as have reflected in his various couplets “Jin khoja tin paiya, gahre pani paith” or “chalti chaki dekh kar diya kabira roye, do patan ke beech main sabit bacha na koi“. For an ordinary person earth is still flat and he may not know the reasons for change of seasons.

Introduction
Antimicrobial resistance (AMR) is increasingly recognised as a “silent pandemic,” threatening global health and development. In 2019 alone, bacterial AMR was directly responsible for 1.27 million deaths and contributed to nearly 5 million more worldwide. Projections warn that by 2050, unchecked AMR could result in 10 million annual deaths and impose a staggering economic burden of USD 100 trillion. A major driver of this crisis is the indiscriminate and often prophylactic use of antibiotics in livestock production systems, which account for a majority of global antimicrobial consumption. The One Health framework—emphasising the interconnectedness of animal, human, and environmental health—offers a strategic pathway for mitigating AMR. It requires dismantling sectoral silos and fostering collaboration among veterinarians, physicians, public health authorities, food safety regulators, and environmental scientists. Against this backdrop, NDDB has positioned itself as a frontrunner in India by embedding One Health principles into large-scale field programmes. Since 2014, NDDB has progressively promoted alternative medicine for bovine mastitis control under its Mastitis Conrol Project (MCP), subsequently expanding in 2017–18 into a broader Disease Control through Alternate Methods (DCAM) project with Ethnoveterinary Medicine (EVM) as the central pillar.
Building Up: From DCAM to DISHA as One Health Strategy
Building on the successes of MCP, DCAM, and BCP, NDDB launched the Dairy Integrated Safety and Health Action (DISHA) project for 2025–2030, DISHA consolidates NDDB’s One Health strategy by combining animal health management, zoonotic disease surveillance, and food safety monitoring. Its objectives include: managing bovine cases with EVM, organising farmer awareness programmes, training veterinarians and health workers, screening cattle as well as their owner for brucellosis, and testing bulk milk samples for aflatoxins and antibiotic residues. Importantly, DISHA also addresses emerging food system contaminants such as microplastics, per- and polyfluoroalkyl substances (PFAs), Bisphenol-A (BPA) and heavy metals, thereby broadening the scope of One Health beyond infectious diseases.

Nocardia has the ability to form gram-positive, branching filaments of less than 1um in diameter. It is closely related to Corynebacterium, Mycobacterium and Rhodococcus species
History Nocard described this organism in 1888, following its isolation from a case of bovine ‘farcy’, hence the name of the type species: Nocardia farcinica.
Natural Habitat
Nocardia species are soil borne saprophytes
Morphology
They are gram-positive, obligate aerobes ability to form branching filaments. Some produce true mycelia and some strains are acid-fast. All species are nonmotile. Gram stained smears from lesions revealed gram-positive branching filaments that often show fragmentation into coccobacillary elements. The modified ZN stained smears exhibit a similar morphology but most of the filaments retain the carbol-fuchsin dye and stain red.
Cultural Characters
The Nocardia species grow very well in blood agar incubated aerobically at 370C for upto 7 days. Inoculate the suspected colonies from blood agar into Sabouraud dextrose agar (SDA) and incubated at 370C for upto 10 days.

Normal bacterial flora is the bacteria which live on another living being (human or animals) or inanimate object without causing infection

Opportunistic bacteria are part of normal bacterial flora which can produce infection during lowered immunity (immunosuppression) of host or when transferred into other parts of the body

Saprophytic bacteria live on dead or decaying organic matter Infection

Infection can be defined as a pathological condition occurred due to multiplication of microbes.

Intoxication can be defined as pathological condition due to ingestion of microbial toxin (e.g. aflatoxin produced by Aspergillus fungus).

Mycetism is the pathological condition due to ingestion of fungus (e.g. toxic mushroom) which produces toxicity within the body of the host

Nucleic acid extraction from clinical samples is the first step to be followed in molecular detection of cancer. Nucleic acids are the starting material for all downstream molecular diagnostic tools. Quality of test results largely depends upon the quality of starting material. Thus utmost importance should be given to the extraction protocols to isolate good quantity and good quality nucleic acids from patient samples.

Basic principle: The basic principle of nucleic acid extraction is to disrupt cell membrane and nuclear membrane to release and precipitate nucleic acid in the solution. Following precipitation,removal of other cellular contaminates such as protein, lipids, polysaccharides etc is performed to obtain a good quantity and quality of nucleic acid.

Introduction
It is reasonable to consider that the history of humanity has been marked by two transformational revolutions: the cognitive revolution (35,000 to 70,000 years ago), when Homo sapiens began to manifest our capacity to abstract, imagine things that did not exist and give meaning to non-representational thoughts; the agricultural revolution (10,000 to 12,000 years ago), when Homo sapiens stopped being a hunter/collector to settle on land, cultivating the soil and raising domesticated animals. It was possibly the beginning of property ownership, monogamy, family and also when malaria, and other diseases more easily spread in communities, became consolidated as human diseases. It was with the cognitive revolution that things changed and our previously acquired abilities of creating images and dealing with abstract concepts started to push humankind to become the “Dominant Mammal” controlling all other animals and Earth’s environments. Progressively, man learned to work with wood, iron and all minerals extracted from the soil, produced clothes with leather and wool from animals, linen and cotton from plants, silk from insects, and latter from natural oils created arts, jewellery, dynamite, electricity, vaccines, transatlantic ships, locomotives, cars, computers, antibiotics, contraceptives, robots, portable cell phones, informatics, bioengineering, internet, artificial intelligence, social networks and many other innovations . Not surprisingly, conscious of all these attributes, man felt important, central and essential, falling into the temptation of judging himself a magnificent creature with all rights and that became deep rooted in some cultures. For the pleasure,comfort, sophistication and power, man interfered with and altered the environment in a so ambitious and predatory manner that he modified and compromised his and other species living conditions, causing also climate changes that threaten his own survival on the planet. With time, it became clear that, in addition to this entirely anthropocentric “Ego’s” point of view, there might be another perspective fromwhich one can perceive the world and interact with it. An ecological “Eco’s” perspective corresponds to a compassionate view (how can we contribute to the greater good and make the world a better place) of the Globe. After all the damage done to nature by man, moving from the socalled Ego to the Eco attitude became a mandatory needConcerning Global Health, for a long-time, human health has benefited by sacrificing the “health” of wild ecosystems (e.g., deforestation and conversion of wilderness to farmland, damming of water for irrigation, destruction of swamps and dislocation and jeopardy of wild species) which reflects a protectionist vs utilitarian conflict, over the question of whether to put human domination of the biosphere on hold or whether to embrace it .It hides unaddressed concerns about the value of (macroscopic) life (human, animal or plant), the definitions of health and wellbeing (human, animal, plant and environmental) and their relative importance. This has hampered the development and acceptance of a One Health (OH) understanding of Global Health.

Microbiologists are constantly at risk of exposure to hazardous microorganisms that can cause diseases. These microorganisms may enter the body through inhalation, ingestion, cuts, abrasions, or the eyes. Therefore, handling cultures, slides, and live microorganisms requires utmost care. To ensure safety in the laboratory, several precautions must be followed:
1. Maintain cleanliness in microbiological labs at all times, taking extra care to prevent contamination from the air, equipment, personnel, or the environment.
2. Always wear a clean white lab coat or apron while working. After handling infectious microorganisms, the lab coat should be autoclaved.
3. No eating, drinking, or smoking is allowed in the laboratory.
4. Personal hygiene is essential in microbiological labs.
5. Wash hands with soap and water or a mild disinfectant solution (e.g., hypochlorite solution containing 50-100 ppm chlorine or 1-2% lysol solution) before starting work.
6. Disinfect the work surface by wiping down the lab bench with a disinfectant solution, such as 5% phenol or hypochlorite solution containing 200-300 ppm chlorine, both before and after work.
7. Keep the work area neat and organized at all times.
8. Use self-adhesive labels instead of moistening labels with your tongue.
9. Sterilize inoculating loops and needles by heating them in a Bunsen burner flame until red hot before and after use.

An organic acid is an organic compound with acidic properties. The most common organic acids are the carboxylic acids, whose acidity is associated with their carboxyl group –COOH. However, sulphuric acids, containing the group –SO₂OH, are relatively stronger acids. Alcohols, with –OH, may also act as weak acids. Some other groups may also act as weak acids viz. thiol group and phenol group. In biological systems, organic compounds containing these groups are generally classified under organic acids. Some common organic acids are lactic acid, acetic acid, formic acid, citric acid, oxalic acid, uric acid, propionic acid and butyric acid. Organic acids have been used for decades in commercial compound feeds, mostly for feed preservation, for which formic and propionic acids are particularly effective. Since the ban of antibiotic growth promoters in many countries, organic acids have been used increasingly, not only because of their preservative qualities, but also for their nutritive properties. Organic acids and their salts are generally regarded as safe and have been approved by most member states of EU to be used as the feed additives in animal production.

Microbiology is a branch of science dealing with the study of organisms which are too small to be seen clearly by unaided eye. Roughly the organisms with a diameter of 0.1mm or less, which can not be seen by the human eyes unaided, come under the domain of microorganisms.

Thus, the laboratory work dealing with the microorganisms needs special care to avoid infection and infusion for maintaining good health. In fact all microorganisms should be treated as potential pathogens. Thus, for working with microorganisms aseptic techniques must be followed by students as well as by the laboratory assistants and teachers.

Location and Layout
• As the laboratory will be handling materials hazardous to human and animal health, it must be isolated as much as possible from livestock yards, poultry farms and quarantine stations and should not function in or near establishments handling healthy animals, food products for human or animal consumption or near vaccine production laboratories.
• Animal houses, garages, workshops, stores, water tanks, residential accommodation, administrative blocks have to be constructed as per the need of ideal microbiological laboratory. Expansion possibilities should
also be kept in mind. The building can be arranged in 5 main groups:
• Administrative / laboratory rooms
• Decontamination and disposal area
• Animal house facilities
• Workshops / stores/garages and
• Residential accommodation for the staff, hospital, schools, playgrounds, parks, shops etc. • The laboratory building may be rectangular in blocks, arranged in L, T or U shapes and may be of one or several stories. The temperature, wind velocity, rainfall,