eChapters

Q1. The exit of drug from C.S.F. irrespective of its entry is by:

1. Passive diffusion
2. Filtration
3. Facilitated diffusion
4. All of the above.

Q2. The number of half-lives required to reach 99% of steady state concentration is:

1. 3.3
2. 4.4
3. 5.5
4. 6.6

Q3. The receptor concept was first introduced by:

1. J.N. Langely in 1878
2. Simonis in 1964
3. Paul Ehrlich in 1926
4. Wakcsman in 1826

Q1. Alpha naphthyl thiourea is less toxic

1. In an empty stomach
2. With oily food
3. Full stomach
4. Not affected

Q2. A rodenticide that blocks tricarboxylic acid cycle by competitive inhibition of the enzyme aconitase

1. Fluoroacetamide
2. Pindone
3. Chloralose
4. All the above

Q3. ANTU is less toxic in an empty stomach a) In an empty stomach it will irritate the stomach wall resulting in vomiting and throwing away the poison and hence no toxicity.

1. Absorb only with fatty materials
2. Immediate absorption and detoxification
3. All the above
4. None of the above

1. Introduction
 Laboratories form the backbone of research, clinical diagnostics and industrial quality control. Good Laboratory Practices (GLP) serve as the foundation for ensuring the quality, integrity, and reliability of scientific research, particularly in different pharmaceuticals drug testing, environmental studies and chemical testing. Good Laboratory Practices (GLP) in animal husbandry and dairy research play a crucial role in ensuring the accuracy and reproducibility of scientific studies related to livestock management, dairy production animal health and promoting animal welfare. Originally developed to prevent fraud and ensure regulatory compliance, GLP has evolved in response to advancements in technology, increased regulatory demands and the growing human and animals need for sustainability in laboratory operations. Laboratory settings are inherently fluid, with regulatory frameworks and technological progress constantly reshaping the environment. Studies indicate that even minor lapses in quality control can trigger significant inconsistencies in test results, outcomes that can have serious repercussions in both clinical and research domains. Therefore, cultivating a philosophy of ongoing improvement is crucial for addressing these evolving challenges, meeting the rising demand for safe, superior dairy products and embracing innovation. This article, synthesize the latest research findings and guidelines, delve into trusted best practices and outline practical strategies for embedding continuous improvement into everyday laboratory operations. A study published in the Journal of Laboratory Quality demonstrated that the adoption of structured quality indicators significantly minimized error rates and bolstered patient safety in clinical laboratories. Their research also provided a comparative analysis of different continuous improvement strategies, including the Plan-Do-Check-Act (PDCA) cycle and Lean Six Sigma, highlighting their respective benefits in lab environments. Based on these insights, the authors advocate for a comprehensive quality management system that seamlessly integrates automation, comprehensive staff training and routine audits to enhance process efficiency. Ultimately, the findings underscore that continuous improvement is an enduring journey, one that evolves along with advances in science, technology and regulatory standards.

Q1. Halothane is metabolized in to

1. Trifluoracetic acid
2. Inorganic chloride
3. Bromine
4. All the above

Q2. Psychomotor stimulants causes

1. Excitement
2. Relieve fatigue
3. Increase motor activity
4. All the above

Q3. Repeated administration of barbiturate causes

1. Tolerance
2. Drug dependence
3. Toxicity
4. All the above

Introduction

For proper and efficient experiment to be conducted, experimenters need so many chemicals to deal with. This includes different acids, base, salt, different kind of indicators and so on. Some of these chemicals are safe for health, but some of them are dangerous to health when come in close proximity to our body while some of the chemicals cause health issues on inhaling. These may cause improper functioning of some of our physiological activities. Acids and bases are very dangerous for our tissues when come in our contact. These may cause serious respiratory problem in addition to injuries on coming to our contact. Some chemicals are exploratory yet they may cause even death of the person working with them in laboratory. Therefore, mere use of the chemicals in laboratory is not only the objective of the experimenter. The person working in laboratory should also be concerned about the proper use and handing of the chemical while working with it. Thus, every researcher of an institution who has to work with chemicals in a laboratory should be made aware about the chemicals to be used in the laboratory and the consequences if any mishandling of a laboratory chemical occurs. Not only these, suitable precautionary measures should also be known to the researcher along with ensuring the availability of the required safety equipment/installations. Hence, a proper understanding of the ways of handling of chemicals in a laboratory is of prime importance for the personals like you who are working or have to work in laboratory.

Safe Handling of Chemicals in Laboratory

Safe use/handling of chemicals in laboratory include minimized exposure to chemicals, maintaining safety data sheets, proper labelling, storage, segregation, and transport of chemicals for personal hygiene.

Since mouth pipetting of liquid reagents in a laboratory is not advisable, different tools are needed for safe, easier, and precise dispensing of liquid reagents. This chapter is devoted to make you aware of the tools like vacupette and micropipette which can be used for the handling of liquid reagents in a safe and precise way.

Vacupette

Glass pipettes have been traditionally used by sucking liquid solvent/solution directly using mouth. For safety reasons, mouth pipetting is no longer permitted in a laboratory. Hence, to fill a pipette vacupette is used by placing it on the top of glass or plastic pipette. It acts as a device to create vacuum for filling reagent in the pipette as well as for releasing the reagent with mechanical regulation on the flow out of the liquid. Vacupette can be of different types.

1. Rubber Bulb Vacupette

Rubber bulb vacupette is often used to transfer (pump up and discharge) liquid using a pipette when a larger quantity of chemicals is to be transferred. Small/tiny rubber bulb is attached to glass/plastic pipette for drawing little amount of reagents. Small vacupette is used to precisely discharge smaller volume/ drops.

Handling of Vacupette

• Insert the top of a pipette in the bottom of the rubber bulb vacupette (Figure 6.1).

• Remove air from the pipette by squeezing valve “A” at the top of the vacupette while simultaneously squeezing the rubber bulb. The amount of air released depends on the size of the rubber bulb.

Introduction

In a research laboratory, we frequently need weigh chemicals and prepare different solutions of varying strength, as per the requirements. Accuracy in weighing and preparation of solution, with respect to weight/volume and strength, determines the success of experiment. In turn, the accuracy depends on weighing of solid chemical and/or measuring the volume of liquid chemical/making up the volume. In this Chapter, weighing of chemicals using appropriate weighing balance, preparation of solutions of different strength, their dilution, and precautions while preparing will be discuss in brief.

Weighing of Chemical

The primary method to determine the amount of chemicals to be used is weighing the chemical by using a balance.

In general, there are two types of balances to weigh chemicals in a laboratory:

1. Standard laboratory balance

2. Analytical balance

Standard Laboratory Balance

Generally, a standard laboratory balance is used for weighing a chemical when an error of ±0.02 g is acceptable. It is also used for pre-weighing, where approximate mass needs to be determined. Precise mass of the chemical is then determined using an analytical balance. Following are the steps in using a weighing balance in a laboratory.

Introduction

Today, antimicrobial resistance (AMR) has become a global issue, contributing to rise in mortality and cost of treatment. Antimicrobial resistance (AMR) is the resistance of a microorganism against an antimicrobial agent to which it was originally sensitive (WHO, 2013). Dispersion of antibiotics is done at large scale, in India even sometimes without any proper prescription. Animal Husbandry is not untouched and antibiotics are used to a great extent, which needs enforcement of strict government rules and regulations for their use in f ield conditions (PHFI, 2011).

Extended-spectrum β-lactamases (ESBLs) are enzymes that hydrolyse most of the β-lactam antibiotics and thus mediate resistance to penicillins, 3rd and 4th generation cephalosporins (Saravanan et al., 2018). Beta-lactamase inhibitors, inhibit these ESBLs such as clavulanic acid, sulbactam and tazobactam (Trott, 2013). The genes encoding for these enzymes are commonly found both on the chromosomes and plasmids among Enterobacteriaceae family (Kotsoana et al., 2019).

Among Enterobacteriacae, E. coli and Klebsiella species are main environmental pathogens associated with various illnesses such as acute bovine mastitis (Koovapra, 2015), pathogenic E. coli with diarrhoea and extra intestinal infections in humans and animals (Levine et al., 1987), K. pneumoniae with respiratory and urinary tract as well as bloodstream infections (Daehre et al., 2018). Resistance to extended-spectrum β-lactams occurs mainly in members of Enterobacteriaceae family through plasmids encoded ESBL genes. (Sharma et al., 2010).

Introduction

Seed is a living entity (mature ovule) consisting of a tiny plant in embryonic stage along with the stored food protected by an outer layer known as seed coat. Seed is an important biological input in agriculture, which is economically important for successful crop husbandry. Cotyledons serve as the source of nutrients (proteins, carbohydrates, and oils) that not only nurture the plant at the early developmental (seedling) stages of plant but also the vegetarian (heterotroph) organisms. Being one of the important inputs in agriculture, viability of seed needs to be assessed to know the germination percentage of seeds used to raise a crop. Since viability of seeds may deteriorate on storage depending on the storage conditions. Therefore, often it becomes important to test viability/germinability and vigour of seeds before being used for raising a crop or conducting an experiment. In this chapter, we briefly discuss about the different methods which can be used for testing the viability of seeds.

Seed Viability

Seed viability can be defined as the capability of the seed to germinate and produce a normal seedling within a specified period. In other words, seed viability indicates how many seeds in a lot are alive and could develop into plants under appropriate field conditions. It is very important that most of the stored seeds are capable of germinating when sown in a field. They must possess higher viability at the beginning of storage and maintain it over the storage. A seed with high initial viability survives longer in storage; however, the viability deteriorates slowly at first and then rapidly as the storage prolongs. It is important to know the duration of storage after which a considerable decline in viability occurs under given storage conditions so that to take necessary action to revive the seeds. Excessive deterioration in vianility of seeds may lead to loss of the seed/material/ germplasm.

Antibiotics, often referred to as “magic bullets” in the battle against bacteria, stand out as one of the most remarkable medical breakthroughs of the 20th century. Their advent revolutionized the therapeutic landscape, consistently saving millions of lives from bacterial infections (WHO, 2020). Undoubtedly, antibiotics are a blessing to humanity, not merely for their medicinal applications but also for their diverse uses. They have long been employed in various contexts, including animal husbandry and production, serving as preventive measures in numerous underdeveloped and developing nations for decades (Majumder et al., 2020). A range of antimicrobial agents, including antibiotics, antifungals, antivirals, disinfectants, and food preservatives, function to either inhibit microbial growth or eradicate them altogether. Among these, antibiotics, specifically tailored to combat bacterial infections, are the most widely used class of antimicrobials. With their increasing and often improper use, microorganisms have developed antimicrobial resistance (AMR) (Murray et al., 2022).

Antimicrobial resistance is an inevitable evolutionary process observed in all microorganisms, and is driven by genetic mutations aimed at resisting the lethal pressures of selection. Bacteria, adapt to environmental selection pressures by developing resistance to antibacterial drugs, thereby diminishing the efficacy of these treatments (Murray et al., 2022). The widespread use of antibiotics, particularly in developing nations, further flares up the process, leading to significant repercussions such as increased rates of morbidity and mortality (WHO, 2020). The incidence and prevalence of antimicrobial resistant bacterial infections have reached alarming levels in the 21st century, posing a substantial threat to global public health akin to a silent pandemic. Antibiotic resistance knows no borders and can impact individuals of any age or gender, regardless of their location. In its present state, antimicrobial resistance (AMR) stands as one of the foremost threats, not only to global health but also to food security (Murray et al., 2022).

1. Introduction

Our Indian traditional medicine depends on herbals which always played an important role in the health problems of many countries. In India, the tribal people are using variety of herbals for effective curing of various ailments. These plants are often used by the local people of that area for treatment of various diseases. Ethnomedicinal studies have offered immense value and opportunities for the development of new drugs. Living close to nature, traditional societies have acquired unique knowledge about the use of wild flora and fauna, most of which are unknown to the people who live away from such natural ecosystem as forests and hills. The hills have been the sources of medicinal plants since the time man realized the preventive and curative properties of plants and started using them for human health care.

Tribal people throughout the world, including India, have developed their own cultures, customs, religious rites, taboos, legends and myths, folk tales, medicine, food, etc., They are the repository of accumulated experience and knowledge of the indigenous vegetation; this can be utilized in tribal development. These days greater emphasis is being laid on this traditional knowledge and on using it in the “bioprospecting” of natural products as a new source of food and medicine.

Antimicrobial resistance (AMR) is a mounting global health emergency that threatens the effectiveness of modern medicine and veterinary care. In India, the challenge is compounded by high antibiotic consumption, unregulated access to antimicrobials, and widespread use in agriculture and animal husbandry. The World Health Organization has identified the Eskape pathogens— Enterococcus faecium, Staphylococcus aureus, Klebsiellapneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp, as priority organisms due to their ability to “escape” the effects of antibiotics and cause life-threatening infections. Simultaneously, zoonotic transmission of resistant bacteria from animals to humans is emerging as a critical concern, particularly in regions with close human-animal interfaces like Jammu & Kashmir. The importance of Eskape pathogens to the establishment and promotion of antimicrobial resistance in hospitalized patients was first recognized in a 2008 by Rice. The morbidity and mortality associated with Gram-negative Eskape pathogens is particularly concerning, as new antimicrobial agentsthose with activity against multidrug-resistant and pan-resistant Gram-negative strainshave not emerged as quickly as needed. Consequently, nosocomial infections continue to pose a serious threat to patient health, especially among critically ill inpatients and those undergoing invasive procedures or requiring the placement of medical devices. Eskape pathogens are well known for their intricate resistance mechanisms, often leaving clinicians with limited therapeutic options. Due to their significant role in driving antimicrobial resistance within hospital settings, consistent and systematic monitoring of their resistance profiles is crucial for guiding effective infection control and therapeutic strategies.This also serves as a critical indicator of regional resistance trends among hospitalized populations. In animal sector, the wastewater of livestock slaughterhouses, animal products (raw or processed) have been considered a source of multidrugresistant bacteria with clinical implications as well as their dissemination into the environment and entry into human food chain.Bacteria from the livestock can act as opportunistic pathogens and carry resistance genes that are important for both veterinary and human health. Since many antibiotics are used in both human and animal health fields, the resistant bacteria enter the environmentespecially through wastewater treatment plants affected by slaughterhouse wasteposing risks to public health. Among the various sources, the water used during poultry slaughtering often contains multidrug-resistant bacteria and can expose slaughterhouse workers to infection. To reduce these risks, it’s important to develop strategies that limit bacterial release into the water systems and prevent their spread to people and the environment. The development of AMR has been attributed to several factors comprising of microbial mutations rendering the antibiotics ineffective, human interference such as over use and over-prescription of antimicrobials, agricultural and commercial application of antimicrobials in the animal sector, and human behavioural factors (Vijayalaxmi et al., 2021). An expected annual rise of antimicrobial resistance (AMR) by 5–10 % annually has been predicted by the Indian Council of Medical Research (ICMR) Annual Report 2021 due to wide spread abusive use of broad-spectrum antibiotics (ICMR Annual Report 2021). Twelve families of priority drug-resistant bacteria (critical, high and medium) posing great threat to human health in terms of resistance to selected antimicrobials have been identified in priority pathogen list (PPL) of Global Action Plan, World Health Organisation (WHO) to stimulate the research towards the development of new antimicrobials against a particular drug-resistance (WHO,2017). From the Indian perspective, AMR is a grave concern as the Indian population is the highest consumer of antibiotics in the world (10.7 units per person). The poor public health systems, hospital infections, high rate of infectious diseases, easy availability of inexpensive

Introduction
Antimicrobial resistance (AMR) is regarded as a silent pandemic that is responsible for causing the majorityof deaths every year and has become a major healthcare challenge and a significant public health issue. It has been reported that AMR alone has caused more fatalities than road accidents and cancer. It is among the top ten global health threats and has been recognized as a part of the multisector One Health approach. It has been predicted that by the year 2050, the burden of AMR will decrease the gross domestic product (GDP) by 2 to 3.5%, causing a loss of USD100 trillion to the world, with the annual death rate reaching 10 million. India faces one of the world’s highest antimicrobial resistance (AMR) rates, driven by high antibiotic consumption in humans, widespread non-prescription use, and unregulated application in livestock, aquaculture, and agriculture. Surveillance data show alarming resistance in major pathogens E. coli and Klebsiella,which now exhibit very low susceptibility to key antibiotics such as cefotaxime, ciprofloxacin, and carbapenems. Factors exacerbating the burden include inadequate infection prevention, limited laboratory capacity, poor enforcement of regulations, environmental contamination from pharmaceutical and hospital waste, and low public awareness. The combined impact threatens public health, food safety, and the economy. In general terms, AMR is defined as the ability of the microorganisms to withstand the action of an antimicrobial agent,i.e. Antibacterial agents, antifungal agents, potentially toxic metals, and disinfectants. AMR is known to be a natural process that occurs over time due to genetic changes in the pathogens under selective pressure, where a pathogen undergoes mutations and is often linked to indiscriminate use of antimicrobial agents.
Indefinite and improper use of antimicrobial agentshas been a major factor in the emergence of AMR. AMR affects both animals and humans alike, and resistant pathogens pose a challenge in treating infections. One of the crucial factors responsible for AMR is theenvironment, which is regarded as both a vehicle for spreading AMR microbes as well is a culprit of AMR pollutants, which severely impact the ecosystems. AMR is a multifaceted problem and to control it effectively, meticulous measures have been implemented for monitoring and surveillance of AMR in clinical settings, in both human and veterinary fields. However, the environmental surveillance dealing with the environmental AMR has yet to be established. The implementation of comprehensive monitoring of AMR in environment will vastly improve our understanding of the various dissemination routes and propagation of the resistant microbes in environment and their role in spreading AMR. In this chapter, we will discuss the environmental spread of AMR, drivers of AMR in the environment, factors influencing AMR, challenges faced, measures to tackle AMR, and future trends in curbing the burden of AMR.

Introduction
Haemoprotozoan diseases are highly prevalent in tropical and sub-tropical countries owing to the ubiquitous distribution of ticks and blood-sucking flies which are natural vectors for the pathogens. In India, theileriosis, babesiosis and trypanosomosis are major haemoprotozoan diseases and cause substantial economic losses to the livestock industry. Though vaccination of the susceptible stock by attenuated schizont vaccine is available for the control of tropical theileriosis in cattle, chemotherapy is the mainstay for the control of haemoprotozoan diseases. However, the limited available antihaemoprotozoan drugs is being depleted by a combination of two factors viz. emergence of drug resistance and the failure to replace old drugs (de Koning, 2017). Drug resistance against parasites is one of the emerging problems affecting veterinary health sector. Since last few decades, development of drug resistance in haemoprotozoan parasites to several drugs has been reported (Tuvshintulga et al., 2019; Ali et al., 2022). Though, understanding of drug resistance in parasitic protozoa is progressing steadily but possibly not as rapidly as parasites are becoming resistant to drugs. The genetic changes in the target sites, the reduced uptake and increased efflux of drugs, and metabolic regulations are the possible mechanism for the cause of drug resistance (Fairlamb et al., 2016). Anti-protozoan drug resistance has been reported in many economically important protozoa to isometamedium, diminazene aceturate, quinapyramine, suramin, melarsomine dihydrochloride, buparvaquone. The resistance initially evolves at genetic level of parasite with genetic traits that allow them to survive exposure to a drug, which is transferred to the subsequent generation thereby potentially increasing the percentage of resistant parasite that can survive subsequent exposure to the chemical. The emergence of drugresistant parasites has serious implications for treating clinical cases, leading to increased morbidity and mortality among the animals and control of the diseases.

Introduction
 Antibiotic therapy is one of the mainapproaches of modern medicine used tofight bacterial diseases worldwide.Antibiotics comprise of any natural, synthetic, or semi-synthetic substances which interfereswith the growth and development of microbes or have killing effect onmicrobes, specifically bacteria. The availability of these antibiotics for treating various bacterial infectious diseases significantly improved the quality of life of humans as well as animals.The period from1930s to 1960s is considered as the ‘golden era’ of antibiotics during whichmost of the antibiotics available today werediscovered. Antibiotics, once known as the magic bullets of modern-day medicine, are now an “endangered species” facing the danger of extinction due to the worldwide emergence of antibiotic resistance (ABR)/antimicrobial resistance (AMR) among microbes. In general, AMR is the capacity of a microbes (bacteria, fungi, and viruses) to resist the growthinhibitory or killing activity of an antimicrobial beyond its normal susceptibility by altering their physiologyor genetic makeup. AMR has emerged as one of the most critical and complicated global public health threat due to its multi-faceted nature.World Health Organization (WHO) has declared AMR as one of the top 10 global public health threats. The emergence of AMR is closely related to the extensive use and misuse of existingantibiotics in the human and animal fields without properguidelines as well as failure to developor discover new antibiotics. AMR leads to a significant health burden worldwide as it is estimated that bacterial AMR alone attributesto 1.14 million deaths globally in the year 2021 and if left unchecked, it may contribute to 10 million deaths annually by 2050.This prediction was recently updated with the data from 2019 and reported that the deaths of 4.95 million people were related to drug-resistant bacterial infections and 1.27 million deaths were directly attributable to AMR.In addition to the health burden, AMR also increases the healthcare costs and imposes significant indirect economic burdens.Increasing reports of treatment failures in patients with infections caused by multi-, extensive-, and pan-drugresistant bacteriahave been documented in literature from different part of the world. Antibiotics once normally used to treat bacterial infections are no longer effective, and it becomes vital to use “reserve” or “last resort” antimicrobials that are often more expensive. Furthermore, WHO has also emphasized that AMR not only makes infections harder to treat but also jeopardizes ancillary medical procedures such as surgeries and cancer treatments. Food may act as a carrier for the transfer of antimicrobialresistant bacteria (AMRB) and antimicrobial resistance genes (ARGs) to humans.Many scientific reports establish a link between the extensive use of antibiotics duringagricultural production and the increase in AMRamong human pathogens, in which food is one of thepossible transmission routes. Antibiotics are regularly added to feed as additives in intensive animal production systems at sub-therapeutic doses to stimulate growth, improve feed efficiency and avoid infections. The extensive use of antibiotics in human treatment, animal therapy and agricultural use as growth promoters are further causes of bacterial resistance to antibiotics. In recent years, the prevalence of AMR among microorganisms isolated from animal source foods (milk, meat, eggs etc) has increased. The increasing demand for quality animal protein has driven intensive farming, leading to increased antimicrobial usage. In the year 2020, global antimicrobial consumption in food-producing animals was estimated around 99,502 tonnes.Antibiotics are commonly used prophylactically infoodproducing animals and it is projected that by 2030 such usewill increase by approximately 67% globally.Despite efforts to reduce antibiotic use, many countries still rely heavily on antimicrobials for growth promotion and disease prevention. The Food and Drug Administration (FDA) has emphasized the need for better monitoring of antimicrobial use in animals. The emergence and spread of AMR in the food chain is considered as a multisectoral issue, because (i) antibiotics are widely used in livestock production systems (cattle, poultry etc), (ii) AMRB and ARGs can easily spread at each stage of the food production chain, and (iii) these AMRB can cause infections in humans.Thus, the emergence of AMR along the food chain is a major

Introduction
The discovery and widespread use of antibiotics in the mid-20th century marked a transformative era in modern medicine, enabling life-saving interventions in surgery, transplantation, and critical care. Diseases once considered fatal became manageable, and the global burden of infectious diseases significantly declined. However, the remarkable success of antibiotics has been increasingly overshadowed by the rapid rise of antimicrobial resistance (AMR), now recognized by the World Health Organization (WHO) as one of the most serious threats to global health, food security, and sustainable development. Resistant infections already contribute to longer hospital stays, increased healthcare costs, and elevated mortality rates. Alarmingly, if current trends continue, AMR-related deaths are projected to exceed those caused by cancer by 2050.A pivotal yet often underestimated factor in the persistence and spread of AMR is the formation of microbial biofilms. Biofilms are highly organized, surface-attached communities of microorganisms embedded in a self-produced extracellular polymeric substance (EPS) composed of proteins, polysaccharides, lipids, and nucleic acids. This matrix not only anchors microbial population but also confers protection against environmental stressors, immune responses, and antimicrobial agents. Bacteria within biofilms can withstand antibiotic concentrations up to 1000 times higher than their planktonic counterparts due to altered metabolic states, enhanced horizontal gene transfer, and activation of resistance mechanisms such as efflux pumps. Biofilms are implicated in over 80% chronic and device-associated infections, including those caused by ESKAPE pathogensAcinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Staphylococcus aureus. These infections are notoriously difficult to treat and often recur, posing significant clinical challenges. Beyond healthcare settings, biofilm formation in livestock and agricultural environmentsexacerbated using antibiotics as growth promoterscontributes to the emergence of resistant strains that can spread to humans via direct contact, environmental contamination, or the food chain.Understanding the multifaceted resistance mechanisms within biofilms is essential for developing effective containment strategies. These include physical exclusion of antibiotics by the EPS matrix, metabolic dormancy, persister cell formation, quorum sensing-mediated coordination, and stress response activation. Moreover, biofilms often consist of multispecies communities, where interspecies interactions further enhance resilience.In response, antibiofilm agents have emerged as a promising frontier in AMR containment. These include natural compounds (e.g., antimicrobial peptides, essential oils), engineered materials (e.g., nanoparticles, surface coatings), quorum sensing inhibitors, efflux pump blockers, and enzymatic disruptors such as dispersin B and DNases. Ecological approaches like probiotics and positive biofilms offer preventive strategies in agriculture and food systems. Importantly, these agents aim to complement antibiotics by enhancing their efficacy rather than replacing them.Despite challenges in scalability, safety, and regulatory approval, antibiofilm strategies represent a paradigm shift in infection control. Their integration into clinical, veterinary, and environmental practicesaligned with the One Health frameworkis essential for preserving antibiotic effectiveness and mitigating the global AMR crisis.

Introduction
Antimicrobial resistance (AMR) has become a major worldwide health issue, requiring the search for natural, safe, and efficient substitutes for synthetic antibiotics. The ancient medical system known as Ayurveda lists a variety of polyherbal compositions having jwaraghna (antipyretic) and krimighna (antimicrobial) qualities. A traditional decoction preparation that is frequently prescribed for microbial infestations (krimi), wound infections, fevers, and skin conditions called Nimbadi Kwatha.Strong herbs that have been separately shown to have antibacterial and immunomodulatory properties, including Nimba (Azadirachta indica), Guduchi (Tinospora cordifolia), Triphala (Haritaki, Bibhitaki, Amalaki), Vasa (Adhatoda vasica), and Katuki (Picrorhiza kurroa), make up Nimbadi Kwatha. Triphala has antimicrobial and antioxidant qualities, Guduchi boosts host immunity and has antibacterial qualities, and Neem provides broad-spectrum antibacterial, antifungal, and antiviral activity. According to experimental studies, giloy extracts are effective against a variety of pathogens, such as bacteria, viruses, and fungi. It has a distinct antimicrobial activity that involves two mechanisms: it enhances phagocytosis, changes cytokine production, and activates macrophages. In light of antimicrobial resistance (AMR), T. cordifolia is a prospective option for the development of plant-based antimicrobials and integrated therapies. Its use is backed by both modern pharmacological research and ethnomedicinal history for infectious disorders and immune health. Nimbadi Kwatha and its derivatives, including Nimbadi Ubatan and soap formulations, have been shown to have antibacterial properties by modern pharmacological research, which also showed a notable inhibition of bacterial and fungal growth in vitro. These results demonstrate the value of Ayurveda in treating infectious diseases and lend credence to its traditional claims. To close the gap between traditional knowledge and contemporary evidence, a thorough analysis of Nimbadi Kwatha’s antibacterial impact is essential. It also identifies the plant’s potential as a natural remedy for drug-resistant infections.

1. Introduction

Pyrus malus Linn. commonly called as ‘apple’ in English and ‘seb’ in Hindi locally, is a well known tree belonging to family Rosaceae. It is indigenous to Europe and West Asia and grow wild in N.W. Himalayas and also cultivated. The leaves and young aerial parts are useful medicinally1. Leaves contain phloretin which is a natural antibiotic. Crushed leaves are used as temporary treatment for wounds. In Homoeopathy, its use is mentioned for labyrinthine vertigo by Dr. Cooper, in Boericke3.  The plant is a tree growing up to 20 to 40 feet, with spreading branches. The leaves are ovate or oblong ovate, serrate, 2-3’’ long; Petioles ½ to 1’’ in length. The flowers are large, fragrant, pale rose in colour and borne in sub umbellate corymbs. Fruit is a pome and edible.

Chemically the plant is  reported to contain patulin, campesterol, stigmasterol, b- sitosterol, nicotinic acid, phloretin 2- O-glucoside, phlorizin, Gibberellin A 62, melibiose, ostreasterol, 1,28-octacosanediol, pomonic acid and 1,24-tetracosanediol.

1. Introduction

Inflammation have been linked with pathogenesis of many diseases like Cancer, Atheriosclerosis, Neurodegenerative diseases, Diabetes Mellitus, Obesity, Arthritis, Cardiovascular diseases, Parkinson’s disease and other deadly diseases (Lee and Weinblatt, 2001). Inflammation can result to genetic defects and immunoregulation and mechanism defects which lead to tissue damage (Jones, 2006). Anti-inflammatory drug was serendipity discovered without thorough investigation and even not effective in term of controlling inflammation and now record several side effect just like other synthetic drugs, thus, safe biological sources are now been considered. Biological sources for active compounds that have medical importance are increase in recent time (Kaboli, 2001). Also, due to the residual side effects of synthetic compounds that form bulk of the materials used for the production of pharmaceutical products. Aspirin can cause stomach bleeding, acetaminophen can cause liver damage Cox-2 inhbitor Vioxx and Celebrex can cause heart problem and non-steroidal anti-inflammatory drugs (NSAID’s) was reported to contribute to numerous death (Clegg et al., 2006; Wolf et al., 1999; Sing, 1998).

Spleen is an elongated solid organ located on the left side along cranial and ventral abdominal wall. The size of spleen is variable in dogs. The spleen is larger in German Shepherds and Greyhounds. In cats the spleen is smaller and has fixed size. It is parallel to greater curvature of stomach within greater omentum. The position of spleen in dogs is variable as it is only attached by gastrosplenic ligament.