1. Problem of Antibiotic Resistance
& Rational use of antibiotics
Dr. Naser Tadvi
Associate Prof., Pharmacology
2. Objectives
• What is antimicrobial resistance
• Why antibacterial resistance is a concern
• How antibacterials work
• Mechanisms of resistance to antibacterials
• Indian scenario
• NDM-1
• Strategies to contain resistance
• Treatment of some resistant bacterial infections
• Summary
3. Introduction
• Throughout history there has been a continual
battle between human beings and multitude
of micro-organisms that cause infection and
disease
4. History
Nobel Lecture, December 11, 1945
Sir Alexander Fleming
The Nobel Prize in Physiology or Medicine 1945
In his 1945 Nobel Prize lecture, Fleming himself warned of
the danger of resistance –
“It is not difficult to make microbes resistant to
penicillin in the laboratory by exposing them to
concentrations not sufficient to kill them, and the
same thing has occasionally happened in the body…
…and by exposing his microbes to non-lethal
quantities of the drug make them resistant.”
6. Why resistance is a concern
• Resistant organisms lead to treatment failure
• Increased mortality
• Resistant bacteria may spread in Community
• Low level resistance can go undetected
• Added burden on healthcare costs
• Threatens to return to pre-antibiotic era
• Selection pressure
7. Drug Resistance
Drug resistance occurs in :
BACTERIA—ANTIBIOTIC RESISTANCE
Endoparasites
Viruses—Resistance to antiviral drugs
Fungi
Cancer cells
8. Antibiotic Resistance
• The concentration of drug at the site of
infection must inhibit the organism and also
remain below the level that is toxic to human
cells.
GOODMAN & GILMAN'S THE PHARMACOLOGICAL BASIS OF THERAPEUTICS - 11th Ed. (2006)
9. Antibiotic Resistance
Defined as micro-organisms that are not
inhibited by usually achievable systemic
concentration of an antimicrobial agent with
normal dosage schedule and / or fall in the
minimum inhibitory concentration (MIC)
range.
Antibiotic Resistance (DR)
= MIC / MCC > Toxic Plasma Concentration
11. Myths of Antibiotic Resistance
1. Drugs (antibiotics) cause organisms
antibiotic resistant.
2. Antibiotic resistant organisms are
more virulent
12. Truth
• Antibiotics select out the resistant strain
• Faulty use of antibiotics or widespread use of
antibiotics increases the probability of such
selection.
• Antibiotic resistant strains appear to be more
virulent because we cannot kill them or stop
their growth.
15. Antibiotic Resistance
Some microorganisms may ‘born’ resistant,
some ‘achieve’ resistance by mutation or some
have resistance ‘thrust upon them’ by plasmids
Some are born great, some achieve greatness
or some have greatness thrust upon them
16. Intrinsic Resistance
It occurs naturally.
1. Lack target :
• No cell wall; innately resistant to penicillin
2. Innate efflux pumps:
• Drug blocked from entering cell or ↑ export
of drug (does not achieve adequate internal
concentration). Eg. E. coli, P. aeruginosa
3. Drug inactivation:
• Cephalosporinase in Klebsiella
17. Acquired resistance
Mutations
• It refers to the change in DNA structure of the
gene.
• Occurs at a frequency of one per ten million cells.
• Eg.Mycobacterium.tuberculosis,Mycobacterium
lepra , MRSA.
• Often mutants have reduced susceptibility
18. Plasmids
• Extra chromosomal genetic elements can replicate
independently and freely in cytoplasm.
• Plasmids which carry genes resistant ( r-genes) are called R-
plasmids.
• These r-genes can be readily transferred from one R-plasmid to
another plasmid or to chromosome.
• Much of the drug resistance encountered in clinical practice is
plasmid mediated
19. Mechanisms of Resistance Gene Transfer
• Transfer of r-genes from one bacterium to
another
Conjugation
Transduction
Transformation
• Transfer of r-genes between plasmids within
the bacterium
By transposons
By Integrons
20. Transfer of r-genes from one bacterium to another
Conjugation : Main mechanism for spread of resistance
The conjugative plasmids make a connecting tube
between the 2 bacteria through which plasmid itself
can pass.
Transduction : Less common method
The plasmid DNA enclosed in a bacteriophage is
transferred to another bacterium of same species.
Seen in Staphylococci , Streptococci
Transformation : least clinical problem.
Free DNA is picked up from the environment (i.e..
From a cell belonging to closely related or same strain.
21. Mechanisms of Resistance Gene Transfer
Transposons
Transposons are sequences of DNA
that can move around different
positions within the genome of single
cell.
The donor plasmid containing the
Transposons, co-integrate with acceptor
plasmid. They can replicate during
cointegration
Both plasmids then separate and each
contains the r-gene carrying the
transposon.
Eg ; Staphylococci,Enterococci
23. Mechanisms of Resistance Gene Transfer
Integrons
Integron is a large mobile DNA
can spread Multidrug resistance
Each Integron is packed with
multiple gene casettes, each
consisting of a resistance gene
attached to a small recognition site.
These genes encode several
bacterial functions including
resistance and virulence.
They cannot promote self transfer
24. Biochemical mechanisms of antibiotic
resistance
• Prevention of drug accumulation in the bacterium
• Modification/protection of the target site
• Use of alternative pathways for metabolic / growth
requirements
• By producing an enzyme that inactivates the
antibiotic
• Quorum sensing
25. Decreased permeability: Porin Loss
Antibiotics normally enter bacterial cells via porin channels
in the cell wall
Antibiotic
Porin channel
into organism
Cell wall
Interior of organism
26. Decreased permeability: Porin Loss
New porin channels in the bacterial cell wall do not allow
antibiotics to enter the cells
Antibiotic New porin channel
into organism
Cell wall
Interior of organism
27. Efflux pumps
• Cytoplasmic membrane transport proteins.
• Major mechanism for resistance in Tetracyclines.
• Some gram -ve bacteria inhibit the plasmid
mediated synthesis of porin channels ,which
obstructs the influx of hydrophilic Penicillins
eg.ampicillin
ATP Binding Cassette
Major facilitator superfamily
Multidrug and toxic compound exporter
Small multidrug resistance transporters
Resistance-nodulation-division
28. Structurally modified antibiotic target site
Antibiotics normally bind to specific binding proteins on the
bacterial cell surface
Antibiotic
Binding Target site
Cell wall
Interior of organism
29. Structurally modified antibiotic target site
Antibiotics are no longer able to bind to modified binding proteins
on the bacterial cell surface
Antibiotic
Modified target site
Cell wall
Changed site: blocked binding Interior of organism
30. Modification/Protection of the Target site
Resistance resulting from altered target sites :
Target sites Resistant Antibiotics
Ribosomal point mutation Tetracyclines,Macrolides
, Clindamycin
Altered DNA gyrase Fluoroquinolones
Modified penicillin binding Penicillins
proteins (Strepto.pneumonia)
Mutation in DNA dependant Rifampicin
RNA polymerase
(M.tuberculosis)
31. Antibiotic inactivation
Inactivating enzymes target antibiotics
Antibiotic
Enzyme
Binding Target site
Cell wall
Interior of organism
32. Antibiotic inactivation
Enzymes bind to antibiotic molecules
Antibiotic Enzyme
binding
Enzyme Binding Target site
Cell wall
Interior of organism
33. Antibiotic inactivation
Enzymes destroy antibiotics or prevent binding to target sites
Antibiotic Antibiotic altered,
destroyed binding prevented
Antibiotic
Enzyme Target site
Cell wall
Interior of organism
34. By producing enzymes that inactivates antibiotic
a)Inactivation of b-lactam antibiotics
•S. aureus, N. gonorrohoea, H.influenza, Produce b-
lactamase which cleaves -lactam ring
b)Inactivation of Chloramphenicol
• Inactivated by chloramphenicol acetyltransferase .
• Gram-ve (enzyme present constitutively hence higher
resistance) gram +ve bacteria (enzyme is inducible )
c)Inactivation of Aminoglycosides
• Inactivated by acetyl, phospho & adenylyl transferases
Present in gram +ve and gram –ve .
35. Use of alternative pathways for metabolic / growth
requirements
• Resistance can also occur by alternate
pathway that bypasses the reaction inhibited
by the antibiotic.
• Sulfonamide resistance can occur from
overproduction of PABA
36. Drug Mechanism of resistance
Pencillins & B Lactamase cleavage of the Blactam ring
Cephalosporiins
Aminoglycosides Modification by phosphorylating, adenylating
and acetylating enzymes
Chloramphenicol Modification by acetylytion
Erythromycin Change in receptor by methylation of r RNA
Tetracycline Reduced uptake / increased export
Active export out of the cell & reduced affinity
Sulfonamides of enzymes
37. Quorum sensing
• Microbes communicate with each other and
exchange signaling chemicals (Autoinducers)
• These autoinducers allow bacterial population
to coordinate gene expression for virulence,
conjugation, apoptosis, mobility and
resistance
38. Why named quorum sensing
• Single autoinducer from single microbe is
incapable of inducing any such change
• But when its colony reaches a critical density
(quorum), threshold of autoinduction is
reached and gene expression starts
• QS signal molecules AHL, AIP, AI-2 & AI-3 have
been identified in Gm-ve bacteria
• AI-2 QS –system is shared by GM+ve bacteria
also
39. WHY INHIBIT QUORUM SENSING
Proved to be very potent method for bacterial virulence
inhibition.
Several QS inhibitors molecules has been synthesized which
include AHL, AIP, and AI-2 analogues
QS inhibitors have been synthesized and have been isolated
from several natural extracts such as garlic extract.
QS inhibitors have shown to be potent virulence inhibitor
both in in-vitro and in-vivo,using infection animal models.
41. Indian scenario
• Lack of community awareness
• Availability over the counter
• Absence of central monitoring agency
• In infants LRTI has taken over IMR due to
diarrhoeal diseases due to use of ORT
• S. Pneumoniae fully resistant to cotrimoxazole
• Still sensitive to penicillins, macrolides and
fluoroquinolones
43. STD
• Penicillin and fluoroquinolone resistance is
widespread to gonorhhoea
• Alternate drugs like Azithromycin and
cephalosporins should be used
• Syphilis still susceptible to Penicillins
44. Gram positive Cocci
• Streptococci other than S. Pneumoniae
– Resistant to tetracycline and macrolides (40%)
– Still sensitive to penicillins
• Staph Aureus
– Methicillin resistance 50%-100%
– Vancomycin resistance also increasing
45. Mycobacteria
• Multidrug resistance
– Combined resistance to rifampicin and isoniazid
• Extensively drug resistant TB
– Additional acquisition of resistance to a
fluroquinolone and one of the three injectable
second line drugs (capreomycin, kanamycin and
amikacin)
• Steady rise in these patients
46. What is NDM-1?
• NDM-1 stands for New Delhi metallo-beta-
lactamase, an enzyme produced by certain
strains of bacteria that have recently acquired
the genetic ability to make this compound.
• The enzyme is active against other compounds
that beta-lactam ring like penicillins,
cephalosporins, and the carbapenems.
• bacteria that produce NDM-1 are resistant to
all commonly used beta-lactam antibiotics,
including carbapenems.
47. New Delhi metallo-beta-lactamase Why
everyone concerned ?
• There are currently no
new drugs in the research
pipelines that aim to stop
NDM-1.To date, some
strains of E.coli and
Klebseilla pneumoniae
are known carriers of the
gene, but the gene can be
transmitted from one
strain of bacteria to
another through
horizontal gene
transfer.
48. Naming the strain as New Delhi creates
controversy
• The gene was named after New Delhi, the capital city
of India, as it was first described by Yong et al. in 2009
in a Swedish national who fell ill with an antibiotic-
resistant bacterial infection that he acquired in India .
The infection was unsuccessfully treated in a New Delhi
hospital and after the patient's repatriation to Sweden,
a carbapenem-resistant Klebsiella pneumoniae strain
bearing the novel gene was identified. The authors
concluded that the new resistance mechanism "clearly
arose in India, but there are few data arising from India
to suggest how widespread it is."
49. Treatment
• Many NDM-1 strains are resistant to all antibiotics
except for colistin.
• Colistin is an older antibiotic that has not been
used much in recent decades, because it is
somewhat more toxic than other antibiotics.
• A few NDM-1 strains have been sensitive to
tigecycline (Tygacil), but this agent should be used
cautiously in serious infections because it does not
achieve high levels in the bloodstream.
• A few strains have also been sensitive to
aztreonam
50. The spread of NDM-1 can be
contained with
..
• The spread of NDM-1 within health-care
facilities can be curbed through strict
infection-control measures, including patient
isolation and hand washing.
51. Strategy to Contain Resistance
• Develop new antibiotics
– Bypass the drug resistance
• Judicious use of the existing
antibiotics:
– Containment of drug resistance
52. New Antibiotic Development
• Only 15 antibiotics of 167 under development
had a new mechanism of action with the
potential to combat of multidrug resistance.
• Lack of incentive for
companies to develop
antibiotics.
54. Hope is not exhausted….yet
• Phage therapy
• Use of the lytic enzymes found in mucus and
saliva
• Agents that target type IIA topoisomerases
• Antimicrobial peptides (AMPs), lipopeptides
(AMLPs) target bacterial membranes,
making it nearly impossible to develop
resistance (bacteria would have to totally
change their membrane composition).
55. Alternate Approaches
Phage therapy
• Phage Therapy is the therapeutic use of lytic bacteriophages to
treat pathogenic bacteria infections
• Bacteriophages are viruses that invade bacterial cells and
disrupt bacterial metabolism and cause the bacterium to lyse.
• Bacteriophage therapy is an important alternative to antibiotics
• The success rate was 80–95% with few gastrointestinal or
allergic side effects. British studies also demonstrated significant
efficacy of phages against Escherichia coli, Acinetobacter spp.,
Pseudomonas spp and Staphylococcus aureus.
Efflux Pump Inhibitors:
56. Some newer antibiotics
• Linezolid: targets 50S ribosome
• Tigecycline: targets 30S ribosome
• Daptomycin: depolarization of bacterial cell
membrane
• Dalbavacin: inhibits cell wall synthesis
• Telavacin: inhibition of cell wall synthesis and
disruption of membrane barrier function
• Ceftibirole/ ceftaroline: cephalosporins
• Iclaprim: inhibits Dihydrofolate reductase
57. Judicious Use of Antibiotics
• Can only contain antibiotic resistance
• Cannot eliminate the possibility of
antibiotic development as resistance is
an evolutionary process
58. Containment of Resistance
• Containment of antibiotic resistance is a
multi-pronged program
• Involves all stake holders
– Physicans
– Patients
– Pharmaceuticals
59. Factors of Antibiotic Resistance
Drug Related Environmental
Factors Factors
Antibiotic
Resistance
Prescriber
Patient Related
Related Factors
Factors
60. 1. Environmental Factors
• Huge populations and overcrowding
• Rapid spread by better transport facility
• Poor sanitation
• Increases community acquired resistance
• Ineffective infection control program
• Widespread use of antibiotics in animal husbandry
and agriculture and as medicated cleansing products
61. 2. Drug Related
• Over the counter availability of antimicrobials
• Counterfeit and substandard drug causing sub-
optimal blood concentration
• Irrational fixed dose combination of
antimicrobials
Policy
• Soaring use of antibiotics Decision at
Higher level
62. 3. Patient Related
• Poor adherence of dosage Regimens
• Poverty
• Lack of sanitation concept
• Lack of education
Patient
• Self-medication
Counseling,
• Misconception Awareness
Program
63. Prescriber Related
• Inappropriate use of available drugs
• Increased empiric poly-antimicrobial use
• Overuse of antimicrobials
• Inadequate dosing
• Lack of current knowledge and training
64. Strategy of Containment
Antibiotic Resistance
Evolutionary Faulty Use of
Process Antibiotics
Hospital Acquired Hospital Environmental
Antibiotic Resistance Community Acquired
Antibiotic Resistance
Empirical Use Definitive Use
Use of antimicrobials before
pathogen responsible for a particular
illness or the susceptibility to a
particular antimicrobial is known
65. Poor Clinical Practice
• Poor clinical practice that fail to incorporate
the pharmacological properties of
antimicrobials amplify the speed of
development of drug resistance.
67. Over Prescribed Antibiotics
• Clinician should first determine whether
antimicrobial therapy is warranted for a given
patient
68. Empirical Microbial Selection
• Is antimicrobial agents indicated on the
basis of clinical findings?
Or is it prudent to wait
until such clinical
findings become
apparent?
72. Empirical Microbial Selection
• What measures should be taken to protect
individuals exposed to the index case to
prevent secondary cases (1), and what
measures should be implemented to prevent
further exposure (2)?
2 1
73. Empirical Microbial Selection
• Is there clinical evidence (e.g. from clinical
trials) that antimicrobial therapy will confer
clinical benefit for the patient?
(Evidence-based medicine)
79. Definitive Treatment
What adjunctive measures can be
undertaken to eradicate infection?
– Vaccination
– Steroid
– Drainage of pus
– Amputation
– Removal of catheter
81. Who’s Work?
Microbiologist
Advise the proper
and adequate
antibiotics with
Physician
balancing the
economy of
hospital Pharmacologist
82. Hospital Acquired Drug Resistance
• Hospital Antibacterial Policy
• Hospital Antibiogram
Hospital specific antibacterial Resistance
Pattern
• Identification of potential pathogen most
likely to cause infection
• Previous antibacterial therapy
• Prescription auditing
83. Hospital Antibiotic Policy
• To curb the common misuse and overuse of
antibiotics
• Restricts the occurrence of antibacterial
resistance among the hospital strains
• Controls the spread of such infections to
susceptible and critically ill patients in the
hospital and the subsequent infection into the
community.
• Saves money for the patient and increases
patient satisfaction with decreased side effect.
84. Hospital Antibiogram
• A periodic summary of antimicrobial
susceptibilities of local bacterial isolates
submitted to the hospital's clinical
microbiology laboratory.
• Used by clinicians to assess local susceptibility
rates, as an aid in selecting empiric antibiotic
therapy, and in monitoring resistance trends
over time within an institution
85. Treatment options for selected highly
resistant bacteria
Sr. Organism Resistance Antibiotic used
No
1 E. Faecalis Penicillin Vancomycin, Ampicillin -SLB
2 MRSA Methicillin etc Linezolid , quinpristine,
Vancomycin dalfopristine , daptomycin,
telavacin
3 S. Epidermidis Methicillin Vancomycin + Rifampicin+
Gentamicin
4 S. Pneumoniae Penicillin G Ceftriaxone, cefotaxime,
Telithromycin
MIC>4 Vancomycin + Rifampicin
5 C. Jejuni FQ Macrolides, doxycycline,
clindamycin
86. Treatment options for selected highly
resistant bacteria
Sr. Organism Resistance Antibiotic used
No
6 E. Coli Cotrimoxazole, Fosfomycin, Nitrofurantoin,
oral Ertapenum
cephalosporins ,
FQ
7 K III Gen Imipenum, Meropenum,
Pneumoniae Cephalosporins Colistin
& Ceftazidime
8 P. aeruginosa Imipenum, Antipseudomal
meropenum Aminoglycosides , Colistin,
Ceftazidime
87. In our hospital antibiotics
Gm +ve bacteria recommended
First line Second line
• Penicillin • Vancomycin
• Oxacillin • Ofloxacin
• Amoxy –Clav • Clindamycin
• Cephalothin • Clarithromycin
• Erythromycin • Linezolid
• Cotrimoxazole
• Ciprofloxacin
• Gentamicin
88. In our hospital antibiotics
Gm -ve bacteria recommended
First line Second line
• Amoxy-clav • Cefta Clav
• Gentamicin • Cefipime
• Ciprofloxacin • Imipenum
• Ceftazidime • Netilimycin
• Cefuroxime • Tobramycin
• Cefazoline
• Amikacin
91. Take Home Message
• Target definitive therapy to known pathogen
• Treat infection, not contamination
• Treat infection, not colonization
• Know when to say “no” to Vancomycin,
Carbepenems and Cephalosporin IV Generation
• Isolate Pathogen
• Break the chain of contagion – Keep your hands
clean.
• Start simple bed side test: Gram stain,
microscopy
Notas del editor
Bubonic plague, TB , Malaria,hiv have affected significant number of hyman beings and caused mortality and morbidity Adult humans contains 1014 cells, only 10% are human – the rest are bacteriaAntibiotic use promotes Darwinian selection of resistant bacterial speciesBacteria have efficient mechanisms of genetic transfer – this spreads resistanceBacteria double every 20 minutes, humans every 30 yearsDevelopment of new antibiotics has slowed – resistant microorganisms are increasing
Antimicrobial agents were viewed as miracle cure when introduced into clinical practice. However it became evident rather soon after the discovery of penicillin that resistance develops quickly terminating the miracle. This serious development is ever present with each new antimicrobial agents and threatens end of antimicrobial area. Today even major class of antibiotics are resistant
If this can be achieved, the microorganism is considered susceptible to the antibiotic. If an inhibitory or bactericidal concentration exceeds that which can be achieved safely in vivo, then the microorganism is considered resistant to that drug. Antibiotic resistance refers to unresponsiveness of microorganism to antimicrobial agents.Susceptible MIC is at a concentration attainable in blood or other appropriate body fluid using usually recommended dosagesResistant MIC is higher than normally attainable levels in body fluidsIntermediate (moderately sensitive, moderately resistant) MIC is between sensitive and resistant levels, may be able to treat with increased dosage
Presence of few mutants not sufficient to produce resistance Single step : E.coli & staph to RifampicinMultistep : erythromycin, tetracyclines, chloramphenicol
The new DNA is then incorporated into the genome of the bacteria which becomes resistant.
They have insertion sequence at end of gene.
They are commonly associates with Transposons.
Commonly operate in E.coli,P.aeruginosa,S.typhi,Staph.aureus,N.gonorroea.
ACYl HOMOSERINE LACTONE,
It remains to be seen if widespread use of antibiotics in syndromic control of LRTI chanfes pattern overtime
Shigella
superbugsgene transfer.
Some antibiotics like aminoglycosides and fluoroquinolones do not contain beta-lactam rings. Unfortunately, the bacteria that have acquired NDM-1 have also acquired other resistance factors and most are already resistant to aminoglycosides and fluoroquinolones. The addition of NDM-1 production has the ability to turn these bacteria into true superbugs (bacteria resistant to usually two or more antibiotics) which are resistant to virtually all commonly used antibiotics.
There are two different approaches to managing antibiotic resistance:1.Managing existing resistant pathogens 2.Avoiding future evolution of more resistanceThe first can be done by, in the case of MRSA, improving hygiene in hospitals, screening hospital visitors and isolating patients
COLONIZATION means that the organism is present in or on the body but is not causing illness.Adopt WHO Strategies and PoliciesVaccination is the most logical and effective means to contain resistance by preventing infection in the first place. For ARIs, diarrhoeal diseases and malaria in children, WHO has developed the Integrated Management of Childhood Illness (IMCI). For the treatment of TB, WHO recommends use of the DOTS strategy. Antimicrobial resistance surveillance – another critical tool in the fight against antimicrobial resistance – identifies and tracks resistance trends in specific infections and geographical locations. INFECTION means that the organism is present and is causing illness.