Antimicrobial

Agents

Dr. Rumita Dey

Introduction
• These are the agents which have lethal or inhibitory effect on the microbes responsible, but in
therapeutic concentration have little or no toxic action on the tissues.
• They are of very diverse chemical structures.
• They can be divided into two categories:
a. Relatively simple compounds obtained by laboratory synthesis, e.g. sulfonamides,
isoniazid, PAS, trimethoprim, etc.
b. Antibiotics are the substances produced by living organisms and which are active
against other living organisms.
• Most of them are produced by soil actinomycetes.
Classification
• Antibacterial agents are divided into two classes on the basis of type of action they exhibit
against bacteria:
1. Bacteriostatic drugs are drugs which, in the concentration attainable in the body,
only inhibit bacterial growth, e.g. chloramphenicol, sulfonamides, tetracyclines, etc.
2. Bacteriocidal drugs are the drugs which kill the bacteria by virtue of their rapid lethal
action, e.g. penicillins, cephalosporins, aminoglycosides, fucidin, nalidixic acid, etc.
Bacteriocidal drugs are more effective therapeutic agents than bacteriostatic drugs.
Mode of Action
• Can be considered from two aspects:
1. Identification of site of action of drug.
2. Its precise mechanism of action.
Loci of action

• There are four major loci of action.

1. Inhibition of synthesis of cell wall
peptidoglycan, e.g. penicillins,
cephalosporin, cycloserine, vancomycin,
ristocetin and bacitracin.
2. Damage to the permeability of the
cytoplasmic membrane, e.g. gramicidin,
polymyxin and antifungal polyene
antibiotics.
3. Inhibition of protein synthesis, e.g. aminoglycosides
(amikacin, netilmicin, tobramycin, gentamicin, kanamycin,
neomycin, streptomycin, etc.), tetracyclines, chloramphenicol.
They bind to and inhibit the function of 30S subunit.

4. Inhibition of nucleic acid synthesis, e.g. rifampicin inhibits

the synthesis of messenger RNA by its action on the RNA
polymerase whereas nalidixic acid inhibits DNA
replication.

Other examples, are novobiocin, pyrimethamine, sulfonamide,

etc.
Mechanism of Action
• There are three general mechanisms of action.
1. Competition with a natural substrate for the active site of
enzyme, e.g.
a. Action of sulfonamides to interfere competitively with the utilization of
paraamino benzoic acid (PABA).
b. b. Action of para-amino benzoic acid with para-amino salicylic acid (PAS).
Mechanism of Action
2. Combination with an enzyme at a site sufficiently close to the
active site as to interfere with its enzymatic function, e.g. vancomycin,
ristocetin and bacitracin.
3. Combination with nonenzymatic structural components, e.g.
drugs which inhibits protein synthesis and drugs which act by damaging
cytoplasmic membrane.
Laboratory Uses of Antibiotics
1. They may be incorporated as selective agents in culture media,
e.g. a) Penicillin may be used for isolation of Hemophilus influenzae
from material taken from upper respiratory tract (penicillin inhibits the growth
of Grampositive bacteria and Neisseriae).
b) Neomycin is used in Willis and Hobb’s medium for the isolation of
clostridia.
Laboratory Uses of Antibiotics
1. For the control of bacterial contamination in tissue cultures used for
virus isolation, e.g. penicillin, streptomycin, nystatin, etc.
2. The pattern of sensitivity of an organism to a battery of antibiotics
constitute a simple method of typing which is of considerable
epidemiological value.
Antibiotic Sensitivity Tests

• Drug sensitivity tests are also important in studies of the epidemiology of

resistance and in studies of new antimicrobial agents.
• MuellerHinton agar media (4 mm thickness plate) is considered best because:
i. Acceptable batch to batch reproducibility for susceptibility testing
ii. It is low in sulfonamide, trimethoprim and tetracycline inhibitors
iii. It gives satisfactory growth of most nonfastidious pathogens
iv. A large body of data and experience has been collected concerning
susceptibility tests performed with the medium.
• This medium should contain as low as possible thymidine or thymine (reverse the
inhibitory effect of sulfonamide and trimethoprim).
• Zones of inhibition are measured to the nearest whole millimeter using sliding
calipers, ruler or template prepared for this purpose which is held on the back of
inverted petri plate.
• These are applied to determine the susceptibility of pathogenic bacteria to
antibiotics to be used in treatment.
Types of Antibiotic Sensitivity Tests

• Mainly sensitivity test are of three types:

1. Diffusion tests: The principle of it is to allow the drug to diffuse through a solid
medium, concentration of drug being highest near the site of application of drug
and decreasing with distance.
There are many methods for implementation of this diffusion test.
The most common, simple and easy method is to use filter paper disks
impregnated with antibiotics (disk diffusion method).
Here filter paper disks 6 mm in diameter are charged with required concentration
of drugs and are stored dry in the cold.
After drying the plate at 37°C for ½ hour antibiotic discs are
applied with sterilized forceps.
After overnight incubation at 37°C, zone of inhibition of growth
around each antibiotic disc is noted.
Inhibition zone shows degree of sensitivity of antibiotic for that
particular bacteria.
The results are reported as sensitive or resistant.
Inoculation of pure bacterial growth in liquid medium,
may be done by spreading with swabs on solid medium.
Disk diffusion test is done only after the pathogenic
bacteria are isolated from clinical specimen in pure form.
Sensitivity tests should be done only with pathogenic
bacteria and not with commensals.
Further, nitrofurantoin need to be tested only against
urinary pathogens.
2. Dilution tests: These are quite laborious for routine use.
• However, these are useful where therapeutic dose is to be
regulated accurately, e.g. in treatment of bacterial
endocarditis and to find out small degree of resistance in
slow growing bacteria like tubercle bacilli.
• In dilution test, serial dilutions of drug are prepared and are
inoculated with test bacterium.
• It may be done by tube dilution or agar dilution methods.
• 3. E. test: It is known as epsilometer test.
Here antibiotic with known gradient of concentration in
length on absorbent strip may be used.
Now above mentioned strip is kept on the petridish
containing nutrient agar seeded with test micro-
organism.
The antibiotic diffuses into nutrient agar medium.
The minimum inhibitory concentration (MIC) is noted
by recording the lowest concentration of the gradient
which, in fact, inhibits growth of microorganisms.
ANTIMICROBIAL RESISTANCE

• Refers to development of resistance to an antimicrobial drug by a

microorganism.

• Can be of two types:

Intrinsic resistance.
Acquired resistance.
Acquired Resistance

• Emergence of resistance in bacteria that are ordinarily

susceptible, by acquiring the genes coding for resistance

• Overuse and misuse of antimicrobial agents is the single most

important cause of development of acquired resistance.

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Acquired Resistance (Cont..)

• Use of the particular antibiotic poses selective pressure in a population

which in turn promotes resistant bacteria to thrive and the susceptible
bacteria to die off.

• Resistant bacterial populations flourish in areas of high

antimicrobial use, where they enjoy a selective advantage over
susceptible populations.

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Acquired Resistance (Cont..)
Factors favoring the spread of antimicrobial resistance include:
• Poor infection control practices in hospitals
• Inadequate sanitary conditions
• Inappropriate food-handling
• Irrational use of antibiotics by doctors
• Uncontrolled sale of antibiotics over the counters without prescription.

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Acquired Resistance (Cont..)

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Intrinsic Resistance

• Refers to innate ability of a bacterium to resist a class of antimicrobial

agents due to its inherent structural or functional characteristics.

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THANK YOU