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The rise in the incidence of infections is caused by multi drugresistant (MDR) bacteria, it is essential to elucidate the basic mechanism ofantibiotic resistance to discover effective methods for diagnosis and treatmentof infections. The use of pathogen-specific probes offers a faster alternative forpathogen detection and could improve the diagnosis of infection. High resolutionmelting analysis techniques are useful for the detection of multi drugresistant pathogens. Rational Structural Based Drug Design is a common methodto identify a lead compound and take it forward for further developments.This book provides information about recent strategies involved in thediagnosis and treatment of infections caused by MDR bacteria. The volume coversthe use of molecular probes for the quantification of pathogenic bacteria, alongwith other techniques mentioned above. Chapters also cover the use of identificationof novel drug targets from the Lipid A biosynthesis and also from quorum sensingmediated biofilm formation in MDR bacteria. Chapters also cover herbal alternatives for the treatment of MDRbacteria like the use of Cassiaaungustifolia in treatment of various diseases. The reference is suitablefor biomedical students, cellular and molecular biologists.

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Table of Contents
BENTHAM SCIENCE PUBLISHERS LTD.
End User License Agreement (for non-institutional, personal use)
Usage Rules:
Disclaimer:
Limitation of Liability:
General:
PREFACE
List of Contributors
Introduction
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Development of Drug Resistance
Abstract
INTRODUCTION
CONCLUSION
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
Acknowledgements
REFERENCES
Current Challenges in Treating MDR
Abstract
INTRODUCTION
Natural/Intrinsic Resistance
Acquired Resistance
MECHANISMS OF MDR DEVELOPMENT
Alteration of the Target Protein by Mutation
Modifications of the Drug
Decreased Antibiotic Penetration and Efflux
Gene Acquisition from other Species for Less Susceptible Target Proteins
Target Bypassing
PREVENTING MDR
MDR Preventive Measures
Political Commitment
Case Finding
Standardized and Supervised Drug Treatment Course
Surveillance System
Development of Novel Drugs
STRATEGIES TO COMBAT MDR
Antimicrobial Peptides (AMPs)
Anti-virulence Compounds
Phage Therapy Alone or in Combinations with Antibiotics
New Molecules
Drug Repurposing and Repositioning
Hand Hygiene
Patient Placement and Priority for Isolation
Personal Protective Equipment (PPE)
Cleaning of the Environment and Patient Care Equipment
Patient Movement and Transfer
Education of Patients, Staff and Visitors
CHALLENGES
Challenges in Diagnosis
Disease Transmission
Treatment of Latent Infection
WHO’s RESPONSE TO MDR
CONCLUSION
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
List of Abbreviations
REFERENCES
Methods of Detection of MDR
Abstract
INTRODUCTION
Techniques to Detect MDR
Culture
Liquid Culture-based Methods
Advantages of Liquid-culture Method
Solid-culture Methods
Advantages of Solid-culture
Polymerase Chain Reaction
Advantages
Disadvantages
Lateral Flow Immunoassay
Advantages
Disadvantages
DNA Microarray
Advantages
Disadvantages
Whole Genome Sequencing
Advantages
Disadvantages
CONCLUSIONS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Molecular Probes as Diagnostic Tools
Abstract
INTRODUCTION
MOLECULAR DIAGNOSTICS FOR DETECTION OF DRUG-RESISTANT PATHOGENS
Polymerase Chain Reactions (PCR)
Real Time Polymerase Chain Reactions (PCR)
Detection Chemistries
Nucleic Acid Dye: SYBR Green I
Sequence-specific Probe
Molecular Beacon (MB)
CONCLUSION
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Real-Time PCR High-Resolution Melting Analysis
Abstract
INTRODUCTION
Principle of the Melting Profile
Intercalating Dyes
Non-saturating Dyes
Saturating Dyes
Release-on-demand Dyes
Working of PCR-HRM
Analysis of Data
Factors Affecting HRM
APPLICATIONS
Identification of Microbial Species
Mutation Detection
Microbial Genotyping within Species
Identification of Genotypic Variants Leading to Antimicrobial Resistance
Identification of Human Genetic Variants of Importance to Susceptibility to or Treatment of Infectious Diseases
Alternative for Gel Electrophoresis
CONCLUSION
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Current Therapeutic Options and Challenges for MDR
Abstract
INTRODUCTION
TRADITIONAL METHODS USED FOR TREATMENT OF MDR
Major Antibiotics used for Treatment of MDR Bacteria
Carbapenems
Polymyxins
Aminoglycosides
Oxazolidinone
Glycylcycline
Synergistic Approach by Combination of Antibiotics
Bioenhancers
CURRENT TREATMENT METHODS USED FOR TREATING PATIENTS WITH MDR BACTERIAL INFECTIONS
Use of Anti-microbial Peptides as Therapeutic Agent
Anti-Virulence Compounds
Drug Repurposing
Phage Therapy for Treatment of MDR Bacteria
CONCLUSION
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Phytomedicines for Bacterial Infections
Abstract
INTRODUCTION
TREES AND SEED OIL
SEED OIL CHEMICAL COMPOSITION
ANTIMICROBIAL ACTIVITIES IN SEED OIL
CONCLUSION AND FUTURE PROSPECTS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Molecular Mechanisms of Antimicrobial Resistance and New Targets to Address Current Drug Resistance
Abstract
INTRODUCTION
ANTIQUITY OF ANTIBIOTIC EXPLORATION AND CONCOMITANT PROGRESS OF ANTIBIOTIC RESISTANCE
THE SCENARIO OF DRUG RESISTANCE: GLOBAL AND INDIAN PERSPECTIVE
SUPERBUGS AND DRUG RESISTANCE
MOLECULAR MECHANISMS OF THE RESISTANCE
Decreased Antibiotics Permeability
Depravity of Antibacterials by Bacterial Enzymes
Enhanced Antimicrobials Efflux
Alternative Pathway Usage by Bacteria
Drug Target Modification
OTHER FACTORS CAUSING ANTIMICROBIAL DRUG RESISTANCE
Self-medication (Drug Abuse)
Evolution and Transmission of Resistance Genes
Travel, Migration, and Trade
Population Mediated Pollution
Climatic Change
Antibiotic Use in Agriculture and Animals
SURVEILLANCE MECHANISM FOR ANTIMICROBIAL RESISTANCE
SUBDUE ANTIMICROBIAL RESISTANCE
New Targets to Address Current Drug Resistance
Raetz Pathway
Other New Antimicrobial Targets
Implementation of Antimicrobial Stewardship Programs
Non-Prescription Antibiotics
Reducing Antibiotic Use in Agriculture and Animals
Vaccination
Clean and Hygiene
Antibiotic Channel Revitalization
NATIONAL POLICIES TO CONSERVE ANTIBIOTIC EFFECTIVENESS
Recommendations from Infectious Diseases Society of America's (IDSA)
CONCLUSION
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Drug Discovery for MDR
Abstract
INTRODUCTION
Emergence of multidrug resistance in Bacteria
Role of structural biology in drug discovery
X-ray crystallography and Computational methods for ligand screening in drug discovery
Challenges and Strategies for Treatment of Multi drug resistant infections
Antimicrobial Peptides (AMPs)
Phage Therapies
Quorum Sensing (QS) and Efflux Pump Inhibitors
CONCLUSION
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Recent Developments to Fight Multidrug Resistance (MDR) in Protozoa
Abstract
INTRODUCTION
Newly Synthesized Organic Compounds
Drug Discovery from Natural Compound Libraries
Protease Inhibitors
Topoisomerase Inhibitors
Antimicrobial Peptides
Other Natural Protozoa Inhibitors
The Drug Repurposing Approach
Therapeutic Applications of RNA Interference (RNAi)
Computational–Based Strategies for Drug Discovery
CONCLUDING REMARKS
FUTURE PERSPECTIVE
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Current Developments in the Detection and Control of Multi Drug Resistance
Edited by
Sanket Kaushik Amity
Institute of Biotechnology
Amity University Rajasthan
Jaipur- 303002,
India
&
Nagendra Singh 
School of Biotechnology
Gautam Buddha University
Greater Noida- 201308,
India

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PREFACE

The ability of different species of bacteria to resist the antimicrobial agent has become a global problem. Despite the development of so many antibiotic molecules during the last fifty years, increasing antibiotic resistance worldwide may lead to therapeutic dead-end. The disease burden from multidrug-resistant strains of organisms causing AIDS, tuberculosis, gonorrhoea, malaria, influenza, pneumonia, and diarrhoea is being felt in both the developed and the developing countries alike. This book summarizes the emerging trends in the field of antibiotic resistance of various MDR bacterial species.

Current strategies of detection and control of multidrug resistant bacteria provides the clear and recent information regarding strategies involved in diagnosis and treatment of MDR bacteria. Over the past decades, due to the exponential rise in the infections caused by MDR bacteria, it is necessary to elucidate the basic mechanism of antibiotic resistance to find out the effective methods for treatment and control. Primary goal of this book is to elaborate and enhance the theoretical knowledge of bacterial pathogenesis, diagnosis and treatment. It concentrates on further elaboration of current strategies of detection and control of MDR bacteria, Quantification of pathogenic bacteria, High Resolution Melting Analysis techniques, Biosynthesis pathways and Rational structure based drug design for MDR infections.

We have attempted to provide large amount of information in each chapter supplemented with in- depth knowledge of current strategies of detection and control of MDR bacteria. This book enhances the knowledge of Microbiology, Biotechnology and Life sciences in field area of MDR pathogenesis and control. The idea to encompass knowledge spanning from what is known to what is unknown in the writings from the experts in the field will result in a rigorous effort to justify the various topics.

We strongly believe that this book will be reader’s delight providing comprehensive understanding of diagnosis and treatment of MDR bacteria. It will be an ideal resource for student’s better understanding of MDR bacteria biology and useful for biomedical students, cellular and molecular biologists. I hope you will find this book interesting and relevant.

Sanket Kaushik Amity Institute of Biotechnology Amity University Rajasthan Jaipur- 303002, India &Nagendra Singh School of Biotechnology Gautam Buddha University

List of Contributors

Aakanksha KalraDr B. Lal Institute of Biotechnology, Jaipur, 302017, IndiaAmit AlokDivision of Radiation Biodosimetry, Institute of Nuclear Medicine and Allied Sciences, Timarpur, Delhi, 110054, IndiaAnupam JyotiDepartment of Biotechnology, University Institute of aBiotechnology, Chandigarh University, Mohali, Punjab, 140413, IndiaAnurag JyotiAmity University Madhya Pradesh, Maharajpura Dang, Gwalior-474005, M.P., IndiaAyushi KhuranaSchool of Biotechnology, Gautam Buddha University, Greater Noida, IndiaDivyapriya KarthikeyanCentre for Bioseparation Technology (CBST), Vellore Institute of Technology (VIT), Vellore, Tamil Nadu, IndiaEna GuptaAmity Institute of Biotechnology, Amity University Rajasthan, Amity Education Valley, Kant Kalwar, NH-11C, Jaipur-Delhi Highway, Jaipur, IndiaGunjan ShailySchool of Biotechnology, Gautam Buddha University, Greater Noida, IndiaJuhi SaxenaDepartment of Biotechnology, University Institute of aBiotechnology, Chandigarh University, Mohali, Punjab, 140413, IndiaJyoti YadavAmity Institute of Biotechnology, Amity University Rajasthan, Jaipur-303002, IndiaKali Kishore Reddy TetalaCentre for Bio Separation Technology (CBST), Vellore Institute of Technology, Vellore, Tamil Nadu, IndiaKishore K.R. TetalaCentre for Bioseparation Technology (CBST), Vellore Institute of Technology (VIT), Vellore, 632014, Tamil Nadu, IndiaKumar Sambhav VermaAmity Institute of Biotechnology, Amity University Rajasthan, NH 11 C Kant Kalwar, Jaipur, IndiaMonish BisenFaculty of Applied Sciences and Biotechnology, Shoolini University of Biotechnology and Management Sciences, Bajhol, Solan, Himachal Pradesh 173229, IndiaMrinalini RoyAmity Institute of Biotechnology, Amity University Rajasthan, Kant Kalwar, NH-11C, Jaipur-Delhi Highway, Jaipur, IndiaMukesh KumarDepartment of Biophysics, All India Institute of Medical Sciences, New Delhi, New Delhi, IndiaNagendra SinghSchool of Biotechnology, Gautam Buddha University, Greater Noida- 201308, IndiaPriyal SharmaAmity Institute of Biotechnology, Amity University Rajasthan, NH 11 C Kant Kalwar, Jaipur, IndiaPunit KaurDepartment of Biophysics, All India Institute of Medical Sciences, New-Delhi, IndiaRajesh Singh TomarAmity Institute of Biotechnology, Amity University Madhya Pradesh, Maharajpura Dang, Gwalior-474005, M.P., IndiaRavi Ranjan Kumar NirajAmity Institute of Biotechnology, Amity University Rajasthan, Jaipur, 303002, IndiaRavneet ChugAmity Institute of Biotechnology, Amity University Rajasthan, NH 11 C Kant Kalwar, Jaipur, IndiaSakshi PiplaniVaxine Pty Ltd, Warradale, Australia; College of Medicine and Public Health, Flinders University, Adelaide, AustraliaSanjit KumarCentre for Bioseparation Technology (CBST), Vellore Institute of Technology (VIT), Vellore, Tamil Nadu, IndiaSanket KaushikAmity Institute of Biotechnology, Amity University Rajasthan, Jaipur- 303002, IndiaSudhir Kumar PalCentre for Bio Separation Technology (CBST), Vellore Institute of Technology, Vellore, Tamil Nadu, IndiaVaishali GurjarSchool of Biotechnology, Gautam Buddha University, Greater Noida, IndiaVijay Kumar SrivastavaAmity Institute of Biotechnology, Amity University Rajasthan, Amity Education Valley, Kant Kalwar, NH-11C, Jaipur-Delhi Highway, Jaipur, IndiaVijay PalAmity Institute of Biotechnology, Amity University Rajasthan, NH 11 C Kant Kalwar, Jaipur, IndiaVinay SharmaAmity Institute of Biotechnology, Amity University Rajasthan, Jaipur-303002, IndiaVinod Singh GourAmity Institute of Biotechnology, Amity University Rajasthan, NH 11 C Kant Kalwar, Jaipur, India

Introduction

Sanket Kaushik1,Nagendra Singh2,*
1 Amity Institute of Biotechnology, Amity University Rajasthan, Jaipur- 303002, India
2 School of Biotechnology, Gautam Buddha University, Greater Noida- 201308, India
*Corresponding author Nagendra Singh: School of Biotechnology, Gautam Buddha University, Greater Noida- 201308, India; E-mail: [email protected]

Antimicrobial resistance occurs naturally over time due to changes at the genetic level of microorganisms. These microorganisms are present in food, animals, plants, humans, and the environment. Many pathogens can spread from people to people or from animals to humans. Some of the major drivers of the development of antimicrobial resistance are misuse (overuse, inappropriate selection and/or dosing, etc.) of antimicrobial agents, poor infection and disease management, lack of access to clean water, sanitation and hygiene for the people, poor access to affordable and quality drugs and diagnostics, lack of awareness and knowledge about health care, etc.

The global resistance among various pathogenic microbes to antimicrobial drugs is now becoming a serious threat to public health worldwide. The failure of treating microbial diseases is leading to prolonged illness and higher expenditure on healthcare along with a greater risk of life. Today, most infectious agents, including viruses, bacteria, fungi and parasites, have developed Multi-Drug Resistance (MDR), which is responsible for drastic increase in morbidity and mortality. In the last few decades, the administration of antibiotics has drastically increased due to a rapid rate of microbial infections; the latter owed to the emergence of MDR in various microbial strains. MDR is defined as a reduced sensitivity of a microbe to an antimicrobial drug that had been used effectively against the same microbe earlier. Such resistant organisms are able to combat the drug leading to ineffective treatment and increased persistence and spreading of the infection in an individual or population.

Antibiotics are a class of chemotherapeutic agents used in the clinical management of microbial infections. Although antibiotics have been used as magic bullets for curing various bacterial infections, their benefits are diminished due to the global emergence of resistant bacterial strains. The term antibiotics was

first coined by Selman Walksman, an American microbiologist, for compounds that inhibit the growth of microorganisms [1]. The modern era of antibiotics started in 1928 with the discovery of penicillin from the culture of the fungus Penicillium notatum by Alexander Fleming, a British doctor. Although the development of antibiotic resistance is mainly attributed to the imprudent and excessive usage of antibiotics, both biosynthetic antibiotic as well as resistance-conferring genes have been known to have evolved much before the use of antibiotics [2-5]. The natural occurrence of antibiotics in sub- inhibitory amounts acts as signaling molecules for quorum sensing, biofilm formation, and in production of virulence factors. The elucidation of the precise roles and mechanisms of antibiotics still remains to be understood. Antibiotics are secondary metabolites of microorganisms that are produced in a very low concentration during the late stages of the stationary growth phase. The drastically different roles of antibiotics have been explored in natural conditions in comparison to the high doses used for therapeutic purposes against microbes. Nutrient starvation induces microbes to produce a large array of small compounds called parvomes [6]. So far, only a small fraction of parvomes has been characterized as antimicrobial agents.

WHO has reported rapid MDR emergence in E. coli,K. pneumonia, S. pneumonia, Mycobacterium tuberculosis and species of Salmonella and Shigella spp. for a number of antibiotic molecules [7]. Various fungi such as Candida, Aspergillus spp., Cryptococcus neoformans, Trichosporon beigelii, Scopulariopsis spp., and Pseudallescheria boydii have been reported as resistant against drugs such as polyene macrolides, azole derivatives, nucleotide synthesis inhibitors and 1,3-β-glucan synthase inhibitors [7, 8]. An extended exposure to antiviral drugs has made viruses also resistant in immunocompromised people. MDR has been observed in several viruses such as cytomegalovirus (CMV), herpes simplex (HSV), Varicella-Zoster (VZV), human immunodeficiency (HIV), influenza A, hepatitis C (HCV), and hepatitis B (HBV) viruses [9, 10]. MDR emergence has also been reported in several parasites, including Plasmodium spp., Leishmania spp., Entamoeba spp., Trichomonas vaginalis, Schistosoma spp. and Toxoplasma gondii for drugs such as pentavalent antimonials, miltefosine, chloroquine, artemisinin, paromomycin, amphotericin B, atovaquone, sulfadiazine and pyrimethamine [11-19].

Most of the infectious organisms have developed high level of alarming MDR against many commonly used antibiotics, being called “super bugs”. Examples of super bugs include MDR bacteria causing urinary tract infections, skin infections and pneumonia. Resistance is known to be a naturally occurring process. A lot of research is currently being performed in order to gain better understanding of resistance mechanisms, which will further aid in tackling the resistant microbes.

The emergence of MDR is a serious threat to society in the current era, since influenza, HIV, tuberculosis, pneumonia, yeast infections and other diseases are major causes of mortality. As per the WHO’s update, in 2018, 3.5% new cases and 18% old cases of tuberculosis in the world are estimated to present MDR. About 8.5% of MDR cases showed extensive drug resistance (XDR) tuberculosis. Over 40,000 cases of XDR-TB cases are projected to be emerging every year. MDR-TB is caused by bacteria that do not respond to at least two of the most potent drugs, isoniazid and rifampicin. Pneumonia is one of the most common infections globally. MDR Gram negative bacteria (MDR-GNB) are responsible for serious threats to the health care system. The most common Enterobacteriaceae isolated from patients with pneumonia were found to be Klebsiella pneumoniae, Enterobacter spp. and Escherichia coli, which accounted for 12%, 8%, and 7%, respectively, of all bacterial isolates included in a study [20]. Antiretroviral therapy for HIV also faces severe challenges due to the development of resistance. Similarly, malaria parasites have also been observed to be resistant to several drugs such as chloroquine, artemisinin, and pyrimethamine [18].

In recent times, infections by MDR microbes have become a big issue in public healthcare management. The overuse or misuse of antibiotic agents in the last few decades is responsible for the emergence of several MDR strains of bacteria, viruses, fungi and parasites. A rapid increase of severe systemic infections along with the development and spread of resistant pathogens are creating a serious challenge to the current healthcare system. Regrettably, so far, the efforts to develop new drugs have not been sufficient to tackle the emergence of new MDR strains. Along with implementing social awareness programs, there is an urgent need to understand the development of resistance mechanisms to find a therapeutic solution for the MDR microorganisms.

CONSENT FOR PUBLICATION

Not applicable.

CONFLICT OF INTEREST

The authors declare no conflict of interest, financial or otherwise.

ACKNOWLEDGEMENTS

Declared none.

REFERENCES

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Development of Drug Resistance

Divyapriya Karthikeyan1,Mukesh Kumar2,Jyoti Yadav4,Amit Alok3,Kishore K. R. Tetala1,Sanjit Kumar1,*
1 Centre for Bioseparation Technology (CBST), Vellore Institute of Technology (VIT), Vellore 632014, Tamil Nadu, India
2 Department of Biophysics, All India Institute of Medical Sciences, New Delhi, 110029, India
3 Division of Radiation Biodosimetry, Institute of Nuclear Medicine and Allied Sciences, Timarpur, Delhi, 110054, India
4 Amity Institute of Biotechnology, Amity University Rajasthan, Jaipur-303002, India

Abstract

The phenomenon of drug resistance is a widely acknowledged problem in clinics. Drug resistance not only increases the treatment time, but also paves the way for testing the maximum limit of dose tolerance of antibiotics in the patient. There is no escaping of the fact that drug tolerance may remain a perpetual problem and bacteria will keep on evolving as a part of the natural selection process. Therefore, novel drugs targeting the novel mechanism of action could be a proposed solution for this problem. The mechanism of action includes efflux pump, alteration/modification of drug target, enzyme inactivation and prevention of drug penetration. The other thing is to avoid the unnecessary usage of antibiotics so that the bacteria living inside the body do not develop resistance. The places where antibiotics can be bought for human or animal use without a prescription, the emergence and spread of resistance are made worse. Similarly, in countries without standard treatment guidelines, antibiotics are often over-prescribed by health workers and veterinarians and over-used by the public. Therefore, this unregulated overuse of antibiotics may lead to an era where normal infection may become difficult to treat and could lead to mortality. The maintenance of hygiene is a must for everyone and it is the only way to get rid of pathogenic bacteria. So, in this chapter, we summarize recent literature on the development of drug resistance, their mechanism of actions used by microbes to develop antibiotic resistance, factors determining their development by infective agents and the spread of resistant bacteria.

Keywords: Antibiotics, Bacteria, Drug resistance.
*Corresponding author Sanjit Kumar: Centre for Bioseparation Technology (CBST), Vellore Institute of Technology (VIT), Vellore 632014, Tamil Nadu, India; E-mail: [email protected]

INTRODUCTION

Drug resistance means the development of mechanisms to show less or lack of sensitivity to a particular drug or drug family. From a medical point of view, the

increase of resistant bacterial strains is an alarming issue but at the same time, it validates the theory of adaptation in evolution. The first actual antibiotic was first discovered by Sir Alexander Fleming [1], and since then, it has been used throughout the world in medicine and plays an essential role in fighting microorganisms [2]. However, excessive use of antibiotics generated multi-resistant bacteria which pose, in recent years, a serious threat to human life [3]. A newer set of antibiotics is required to treat infections that were treated easily in the past. Bacteria surviving the hospital setup are normally drug-resistant and often require strong antibiotics with prolonged administration. Antibiotic resistance is emerging as one of the global problems to human and animal health. One year of exposure to antibacterial agents also developed resistance in many bacterial species. This is due to the constant evolution of microbes to overcome the antimicrobial compounds produced by other microorganisms and synthetic compounds [4]. The evolutionary history of pathogens reveals that antibiotics have played a pivotal role in their evolution [5]. Several important factors can accelerate the evolution of drug-resistant microbes. These include the sub-therapeutic dosing, misuse and overuse of antimicrobials, and patient noncompliance with the recommended course of treatment. This resistance develops over the due course of time due to survival mechanisms adopted by pathogen against a drug or, to a lesser extent, change in the receptor activity of a host [6, 7]. Extended usage of antibiotics has resulted in the emergence of antibiotic-resistant bacteria, which are often difficult to eradicate requiring either a higher dose or a new class of antibiotics [8].

There are several strategies used by microbes to develop antibiotic resistance which falls into the four categories such as efflux pump, alteration/modification of drug target, enzyme inactivation and prevention of drug penetration (Table 1). Although intrinsic resistance utilises efflux pump, enzyme inactivation and limited uptake of drugs and acquired resistance utilises may be drug target modification, efflux pump and enzymatic inactivation [9].

Table 1Mechanism of drug resistance.S.No.Mechanism of Drug ResistanceDrug Resistant Agents1Efflux pumpFluoroquinolones, aminoglycosides, tetracycline, β-lactams, macrolides2Alteration/modification of drug targetFluoroquinolones, rifamycins, vancomycin, β-lactams, β-lactams, aminoglycosides3Enzyme inactivationβ-lactams, macrolides, β-lactams, macrolides, rifamycins4Prevention of drug penetrationβ-lactams, tetracycline, fluoroquinolones

Antibiotic resistant bacteria can modify their active site where antibiotics act, evolving a novel survival strategy [10, 11]. Researchers have also found several shreds of evidence where these microbes have an ability to inhibit the accumulation of antibiotics, thereby decreasing their effective concentration inside the bacterial cell. Bacteria produce efflux pumps to transport various antibiotic molecules. The efflux pump also flushes out antibiotics and prevent the accumulation of drug, thus, rendering bacterial resistance [12, 13]. Frequent mutations in the bacterial DNA can make the bacteria produce more pump molecules, increasing resistance. Bacteria also can change their membrane permeability. Membranes have pores that allow the transportation of molecules; thus, decreasing the permeability would mean lesser uptake of antibiotics [14, 15].

Bacteria seem to be an advanced version of this theory as some of them have developed an ability to produce enzymes that degrade antibiotics [11, 16]. Penicillin is known as the first discovered antibiotic and certain bacteria produce beta-lactamase enzymes, which degrade the beta-lactam ring of penicillin to inactivate it [17]. Some bacteria produce enzymes that are capable of adding a different chemical group to the antibiotic molecule, preventing binding between the antibiotic and its cell target. For example, Mycobacterium tuberculosis produces a protein that mimics the structure of DNA and this protein binds to fluoroquinolone antibiotics, thus preventing them from binding to the bacterial DNA. This property makes M. tuberculosis resistant to fluoroquinolones, sulphonamides, trimethoprim and DNA gyrase [18, 19].

Microbial resistance can occur in another manner when an antimicrobial drug functions as an antimetabolite by targeting a specific enzyme to inhibit its activity [20, 21]. First, bacteria may overproduce the enzymes, which are the drug targets and these target carry out enzymatic reactions. Secondly, the cell wall of bacteria develop a bypass mechanism which is not susceptible to binding of the antibiotics. Both of these strategies have been found as mechanisms of sulfonamide resistance [22, 23].

Many antibiotics act on a bacterial target (usually a protein), inactivating its activity. Some bacteria produce DNA mutations that modify the target so that these drugs may not bind to it [24]. Some bacteria add different chemical groups to the target, thus, shielding themselves from antibiotics, while some other bacteria express alternative proteins which can be used to inhibit the action of antibiotics. Hence, Staphylococcus aureus can produce a new penicillin-binding protein by acquiring the resistance gene mecA [25].

The targets of β-lactam antibiotics are proteins required for bacterial cell wall synthesis. This type of resistance is the basis of MRSA (methicillin-resistant Staphylococcus aureus). Some bacteria have developed the ability to synthesize a new cell wall altogether for antibiotics [26]. Vancomycin-resistant bacteria have the capability to make up a different cell wall compared to susceptible bacteria. The mechanism of vancomycin-resistant among bacteria involving structural changes in peptidoglycan subunits of bacterial cell wall, prevent vancomycin from binding [22].

Pathogenic bacteria can acquire antibiotic resistance genes via vertical and horizontal gene transmission. Microbial species transfer their resistant gene to progeny and also can transfer genes between species, known as horizontal gene transfer [27-29]. Vertical gene transmission is an evolutionary process which occurs due to errors or mutation in the genome during replication. Horizontal gene transmission is one of the most significant dominating factors determining antimicrobial resistance. The gene responsible for antimicrobial resistance is transported through mobile genetic elements in the process of transformation, transduction, or conjugation [27]. In conjugation, two bacteria connected through structures in the cell wall and transfer DNA from one species to other. In the transduction process viruses, also known as bacteriophage take up the DNA and transfer from one bacterium to another and in transformation process, bacteria share genes by horizontal gene transfer in which bacteria can take up genetic material directly from the environment around the cell [27].

Superinfections, also known as secondary infections, due to the overuse of antibiotics in the hospital environment, have been responsible for the death of more than 5 hundred thousand individuals every year. Biofilm associated infections can also contribute to life threatening diseases such as endocarditis, rhinosinusitis and periodontitis, etc. Mostly biofilm formation can be observed on medical devices like urinary-catheters and medical implants [30]. Thus, preventive measures such as immunization, hygiene, intake of freshly prepared and well-cooked foods, and consulting doctors at the early stage by all individuals are the key elements to decreasing or reversing the worsening situation of antibiotic resistance [31].

In spite of creating much awareness among the public, the problem of antimicrobial resistance is alarmingly high, leading to the evolution of multi-resistant bacteria or “superbugs”. To overcome this, the Longitude Prize has come up with the idea of launching a gaming app named Superbugs, especially for high school students. The aim of this mobile game (made with a combination of both science and fiction) is to make the individual understand how to survive and use the existing and new antimicrobials wisely [32]. Some of the solutions to this growing antimicrobial resistance are approaching new targets, drug repurposing, considering conventional drugs, combination therapy, phage therapy [33], scaling up antibiotic stewardship, reducing antibiotic use in agriculture and livestock, and using multidisciplinary approaches.

CONCLUSION

The development of antibiotic resistance is a major public health concern worldwide. In this chapter, mechanisms of development of drug resistance in bacteria have been studied. There are several mechanisms that have been discussed and are used by the microbes to develop antibiotic resistance such as alteration/modification of drug target, enzyme inactivation, efflux pump and prevention of drug penetration. The novel drugs presenting different mechanisms of action are being developed with the aim of bypassing the drug resistance phenomenon. The resistant bacteria can be transfer genetic material which are responsible for resistant not only to their same species to other bacterial species inhabiting the same environment. Transfer of antibiotic resistance can occurred by Conjugation, Transduction and Transformation. Antibiotic resistance is accelerated by the misuse and overuse of antibiotics. Biofilm formation, poor infection prevention and control can contribute to spreading infection caused by bacteria that possess resistance. Hence, it adds to the emergence of resistance, worsening the problem. There is an urgent need for action to be taken by each level of society to contain infections by resistant bacteria.

CONSENT FOR PUBLICATION

Not applicable.

CONFLICT OF INTEREST

The authors declare no conflict of interest, financial or otherwise.

Acknowledgements

S. K and Kishore acknowledge the financial assistance from the Indian Council of Medical Research (ICMR). DK thanks to ICMR for the senior research fellowship (SRF). MK thanks to ICMR for RA ship.

REFERENCES

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