GENERAL  ARTICLE

Antibiotic Resistance of Bacteria: A Global Challenge

Saswati Sengupta and Madhab K Chattopadhyay

Bacterial resistance to antibiotics poses a serious challenge to

the prospect of chemotherapy. Rational use of antibiotics is
most desirable but it cannot provide a permanent solution to
the problem. In this article the biochemical and genetic basis
of antibiotic resistance in bacteria is discussed with examples. 1 2

The non-clinical aspects of antibiotic-resistance are also dealt

1
Saswati Sengupta is a
with in brief.
reserach associate at the
Indian Institute of
1. The Wonder Drugs Chemical Technology
(CSIR). Her areas of
Antibiotics constitute one of the most significant contributions of interest are therapeutic
modern science. The discovery of these life-saving drugs trans- proteins and intellectual
property rights.
formed the health-care scene during the last century. A signifi-
cant decline in the fatality rate of many diseases was noticed after 2
Madhab K Chattopadhyay
the introduction of antibiotics into clinical practice. For example, is a scientist at the Centre
20 to 85% death rate due to pneumonia in the US during the 1930s for Cellular and Molecular
Biology (CSIR). His areas
came down to about 5% in the 1960s, 100% death due to chronic
of interest are stress
infection of a heart valve was reduced to 5%, 20 to 90% fatality adaptation of bacteria and
due to epidemic of spinal meningitis infection caused by the antibiotic resistance in
bacterium Neisseria meningitides, was decreased to almost 2%. microorganisms.

Antibiotics help not only in curing the diseases but also in their
prevention. Penicillin, for instance, prevents throat infection
caused by Streptococcus and thereby prevents recurrences of
rheumatic fever in susceptible individuals. Onset of a sexually
transmitted disease may be prevented by the timely use of the
proper antibiotic. By eliminating the causative organism of diph-
theria from carriers (people harboring the organism without
falling sick) using an antibiotic, the chances of dissemination of
infection could be effectively reduced. Keywords
Antibiotic-resistance, bacteria,
2. What They Are genetic basis, biochemical ba-
sis, antibiotic paradox, multidrug-
In 1942, Selman Waksman defined antibiotics as low molecular resistant.

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Any chemical weight chemical substances produced by microorganisms, hav-

substance either of ing growth-inhibitory effects on other microorganisms in high
microbial, synthetic dilution. However in the present context, any chemical substance
or semi-synthetic either of microbial, synthetic or semi-synthetic origin, used in the
origin, used in the clinical treatment of infections, is called antibiotic. Thus strepto-
clinical treatment of mycin produced by fermentation is an antibiotic. The sulpha
infections, is called drugs and quinolones, produced by chemical synthesis, are anti-
antibiotic. biotics. Ampicillin (semisynthetic penicillin), produced partly by
fermentation and partly by chemical synthesis, is also an antibi-
otic. The discussion in this article will be confined to the antibac-
terial antibiotics.

The significance of antibiotics in nature remains a riddle. They

are secondary metabolites and therefore are not indispensable for
sustenance of life of the producer organism. The widely- believed
notion about their role in preventing the growth of other organ-
isms which thrive on the same source of food in natural environ-
ments, is essentially anthropocentric and yet to be validated. The
role of antibiotics as signaling molecules in natural ecosystems
has been suggested by some scientists.

3. Historical Background

Though the use of microorganisms for the control of infections

was in vogue even in ancient civilizations, the modern era of
antibiotics was ushered only at the beginning of the twentieth
century. During the screening of hundreds of synthesized organic
arsenical compounds for efficacy in the treatment of syphilis,
salvarsan was discovered by Sahachiro Hata in 1909 in the
laboratory of Paul Ehrlich, the German Nobel-laureate. Prontosil,
the first sulfonamide drug, was developed by Gerhard Domagk in
The role of 1932 for commercial use. The serendipitous discovery of penicil-
antibiotics as lin by Alexander Fleming at St Mary’s Hospital (UK) in 1928 and
signaling molecules the subsequent isolation and purification of the compound by
in natural Florey and Chain made a giant stride in the field of chemotherapy.
ecosystems has Availability of such a compound with activities against a wide
been suggested by range of infectious organisms and (unlike sulfonamides) no loss
some scientists. of activity in the presence of biological materials such as pus,

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provided a distinct advantage to the physicians in controlling Within four years

bacterial infections. Discovery of streptomycin, the antitubercu- following the
lar antibiotic, by Albert Schatz, a graduate student in the labora- introduction of
tory of Selman Waksman (associated with the discovery of a penicillin during the
series of antibiotics in the 1940s and 1950s) at Rutgers University Second World War,
(USA) in 1943, led to the idea that all the infections could be occurrence of
controlled whenever required. Major pharmaceutical companies resistant strains was
started diverting substantial portion of their funds, earmarked for reported.
research on anti-infectives, to the development of other drugs.

4. Emergence of Antibiotic-Resistant Bacteria

The euphoria did not last long. Within four years following the
introduction of penicillin during the Second World War, occur-
rence of resistant strains was reported. According to an estimate
by the The Centers for Disease Control and Prevention (USA),
13,300 patients died of antibiotic-resistant bacterial infection in
the US during 1992. An incredible 150% increase in the occur-
rence of drug-resistant Pneumococci was noted between 1987
and 1994. A 20-fold increase in the frequency of hospital-ac-
quired Enterococci, resistant to vancomycin, was seen between
1989 and 1993. The frequency of methicillin-resistant Staphylo-
coccus rose from 2% in 1975 to 32% in 1992. By this time,
resistance to virtually all the therapeutically useful antibiotics
had been evidenced. Emergence of methicillin-resistant Staphy-
lococcus aureus (MRSA) and vancomycin-resistant Enterococci
(VRE) have raised serious concern all over the world since these
two antibiotics were believed to be invincible when they were
released in the market. A couple of years back, some Gram-
negative bacteria (Escherichia coli, Klebsiella pneumoniae) were
found to produce an enzyme, which confers resistance to virtually
all the commonly used antibiotics including carbapenems, one of
the last resorts in the clinical management of infections caused by
multidrug-resistant organisms. The organisms were believed to
be originated from India. That is why the enzyme was named New
Delhi Metallo -lactamase (NDM-1). Some of the other most
important types of multiple drug-resistant organisms include
extended-spectrum beta-lactamase producers (which are resis-

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1 A process of gene transfer in tant to cephalosporins and monobactams) and penicillin-resistant

bacteria facilitated by direct up-
Streptococcus pneumoniae. Studies on resistance in soil bacteria
take of exogenous genetic ma-
terial (such as plasmid DNA) have been shown that these organisms serve as a reservoir for
from a related strain in the sur- multiple antibiotic resistances. A recent analysis of 264 soil
roundings. Transformation takes isolates obtained from different natural habitats in and around
place naturally in some species
Hyderabad has identified 5 isolates resistant to as many as 10
and can also be attained by arti-
ficial means. This technique is antibiotics. During the recent past, soil metagenomics also re-
being used very frequently in the vealed several aminoglycoside resistances in nonculturable bac-
field of biotechnology for the teria. Notwithstanding the availability of so many antimicrobial
purpose of cloning.
agents, infectious diseases still remain the second leading cause
2 Another process of bacterial
of death worldwide. Eventually, the widespread occurrence of
gene transfer mediated by bac- antibiotic-resistant bacteria has added a new dimension to the
teriophages which carry DNA emerging threat of bioterrorism.
from one cell to another. This
process is known to occur in 5. Genetic and Biochemical Basis of Antibiotic Resistance
several bacterial species such
as Salmonella, Escherichia, Mi- 5.1 Genetic Basis:
crococcus etc.
5.1.1 Spontaneous Mutation and Gene Transfer: Antibiotic-
3 The process which involves
tolerance in bacteria emerges as a result of error in DNA replica-
the transfer of genetic material
from one bacterial cell to an- tion, a phenomenon known as spontaneous mutation having a
other by direct cell-to-cell con- frequency of one in 107 cells. During the epidemic of Shigella
tact through a tube-like struc- infections in Japan during 1950s, it was observed that bacteria
ture known as pilus.
could transfer copies of antibiotic resistance genes to susceptible
4 The movement of genetic ma- bacteria thus making the latter antibiotic-tolerant. The mecha-
terial between unrelated species nism predominantly used for this purpose is conjugation1, a
of bacteria. This can happen powerful gene-mobilizing mechanism. The other known mecha-
through direct incorporation of
nisms of gene transfer among bacteria, e.g., transformation2 and
free DNA by bacterial cells. This
direct form of gene transfer, for transduction3 also play some role in dissemination of antibiotic
instance in the soil or in the resistance. Resistance thus acquired is transferred to the progeny
digestive tract of animals, is the cells by a process called vertical gene transfer. Another process,
most common mode for the
called horizontal gene transfer4, enables bacteria to transfer cop-
transfer of genetic material. It
fosters rapid dissemination of ies of antibiotic resistant genes even to the distantly related
antibiotic resistance in the bac- species in their neighborhood and thus contributes significantly
terial community. Horizontal in dissemination of resistance. Different types of such resistant
gene transfer can happen either
genes, when accumulated in a single cell, result in multi-resistant
through transformation, trans-
duction or conjugation. phenotype.

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5
Mobile genetic elements, viz., transposons5 and integrons6 can Small segments of DNA which
can hop from one DNA location
also transfer antibiotic resistance genes en block to susceptible
to another. It is found as part of
bacteria. Resistance genes mostly reside on bacterial plasmids. a bacterium’s nucleoid (conju-
However, in some cases they are also found to be located on gative transposons) or in plas-
chromosomes and in a few cases organized in operons. It was mids. It contains a number of
genes, coding for antibiotic re-
believed earlier that antibiotic-resistant strains were less compe-
sistance or other traits, flanked
tent to survive in the environment and are slow-growers com- at both ends by insertion se-
pared to their antibiotic-sensitive counterparts, but subsequent quences called transposase,
studies have established that such disadvantages might be coun- which catalyzes the cutting and
resealing of the DNA during
terbalanced by additional compensatory mutations.
transposition. Thus it is able to
cut itself out of a bacterial nucle-
5.2 Biochemical Basis
oid or a plasmid and insert into
another location leading to the
5.2.1 Drug Inactivation by Microbial Enzymes: Among several
transmission of antibiotic resis-
mechanisms involved in the development of antibiotic resistance, tance among a population of
drug modification plays a significant role in rendering many bacteria.
therapeutically useful drugs useless. For example -lactamase,
6 A mobile genetic element,
an enzyme elaborated by many Gram-positive and some Gram-
which contains sitespecific re-
negative bacteria, converts penicillin into penicilloic acid, which combination system capable of
is therapeutically inactive. The production of the enzyme is capturing and mobilising gene
inducible in Gram-positive bacteria whereas it is constitutive in cassettes. It is composed of an
intI gene encoding an integrase,
Gram-negative bacteria. This reaction also forms the basis of
a recombination site attI, and a
chemical assay of the antibiotic since penicilloic acid can be promoter (5’ to the cassette) that
titrated with iodine. The enzyme also provides a therapeutic mediates expression of the re-
target in the management of infections caused by penicillin- sistant determinant. The 3’ end
of the cassette has a variable
resistant bacteria. Likewise, chloramphenicol is converted to the
region termed as 59bp element,
therapeutically inactive compound 1, 3-Diacetoxychloramphenicol which contains the recognition
by chloramphenicol acetyl transferase (CAT), produced by some sequence of the integrase. The
resistant bacteria. N-acetylation of kanamycin by a bacterial integrase is able to integrate or
excise genes-cassettes (usu-
enzyme leads to tolerance of the organism to the antibiotic.
ally antibiotic-resistance genes),
by site-specific recombination.
5.2.2 Modification of Target Site: The other mechanism of
For dissemination, the integron
antibiotic-resistance e.g., modification of the target is best exem- must be able to insert itself into
plified by streptomycin and erythromycin resistance of bacteria. a plasmid or transposon. To date
Both of these antibiotics act by ribosomal binding thereby inhib- integrons are found in Gram-
negative bacteria.
iting bacterial protein synthesis. Modification of the S12 protein
of the 30S subunit of the ribosome makes the ribosome insensi-
tive to streptomycin. Mutations affecting protein L4 or L12 of the

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50S ribosomal subunit render it resistant to erythromycin. An-

other example is the modification of the -subunit of RNA-
polymerase of Mycobacterium leading to the failure of the anti-
tubercular drug rifampicin to bind the subunit and inactivate the
enzyme.

5.2.3 Reduction in Permeability to Antibiotics: In some cases,

emergence of mutants with reduced permeability of the cell
membrane to antibiotics compared to that of the wild-type strain,
leads to tolerance to the antibiotic. For example Neisseria gonor-
rhoea, the causative organism of gonorrhea (one of the most
prevalent sexually transmitted disease), can gain antibiotic resis-
tance by acquiring a mutation in the gene encoding the membrane
protein ‘porin’, thus inhibiting the transport of the antibiotics
penicillin and tetracycline into the cell and rendering the cells
immune to the effect of drugs.

5.2.4 Exclusion of Antibiotics from the Cell: Among the mecha-

nisms involved in the resistance of bacteria to tetracycline, en-
ergy mediated efflux is a powerful strategy, which does not allow
the drug to accumulate in sufficient concentration to exert its
inhibitory effect. It is mediated by a trans-membrane export
protein that functions as an elctroneutral antiport system. The
protein catalyzes the exchange of a tetracycline-divalent metal
cation complex for a proton.

5.2.5 Overproduction of a Target Metabolite: In some cases, the

molecule, which is competitively antagonized by the antibiotic, is
overproduced. For example, sulphonamides act by competitively
inhibiting the enzyme dihydropteroate synthetase, which plays a
crucial role in the synthesis of folate. Sulfonamides (or sulfa
drugs) are structural analogs of p-aminobenzoic acid (PABA), the
substrate of this enzyme. The indispensable role of folate in the
synthesis of nucleic acids viz., DNA and RNA is well-known. In
some PABA-overproducing mutants of Staphylococcus aureus,
sulfonamide molecules are outnumbered by the substrate and
therefore the activity of the enzyme is not inhibited even in the
presence of the drug.

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5.2.6 Gene Deletion: Gene deletion is also known to contribute to

the mechanism of antibiotic resistance. Deletion of the katG gene
in some strains of Mycobaterium tuberculosis is associated with
the tolerance of the organism to the anti-tubercular drug
isonicotinic acid hydrazide (INH). It is postulated that INH is
actually a prodrug [15], which is converted into the active form by
catalase/peroxidase, encoded by katG. Thus, absence of the katG
gene makes the organism immune to the effect of the drug.

6. Adaptive Resistance

In the laboratory, a bacterial population can adapt to tolerate an

antibiotic following exposure to gradually increasing concentra-
tion of the drug. While resistance due to mutation occurs in low
frequency, resistance due to adaptation imparts tolerance to the
whole population at a time. It is usually caused by a change in the
lipid composition of the cell wall ultimately leading to reduced
permeability to the antibiotic. The maintenance of the resistance
phenotype requires the presence of an antibiotic in the growth
medium unlike mutational resistance, which does not require the
presence of the selection pressure. However, with the evidence
available so far, adaptive resistance does not appear to play any
significant role in therapeutics.

A special type of non-heritable resistance to chloramphenicol and

ampicillin was also found to be induced in Escherichia coli when
it was incubated in the presence of repellants like sodium acetate,
acetyl salicylate, salicylate and sodium benzoate. The inducers in
turn made the cells tolerant to ampicillin and nalidixic acid.
Induction of tolerance to structuarally unrelated antibiotics by a
variety of substances including some pharmaceuticals (acetyl
salicylate) is commonly known as phenotypic antibiotic resis-
tance and it appears to have some significance in therapeutics.

7. Non-Specific Resistance

Apart from harboring antibiotic resistance genes in mobile ge-

netic elements such as plasmids and transposons, many bacterial
species possess an intrinsic mechanism for resistance to multiple

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Bacteria can also structurally unrelated compounds. The chromosomal multiple

survive antibiotics antibiotic resistance (mar) locus of some members of the Entero-
by the formation of bacteriaceae confers resistance to several antibiotics, household
biofilms, a multilayer disinfectants, organic solvents and other toxic chemicals. In E
conglomeration of coli, multiple antibiotic resistance is known to be conferred by
diverse-species of different sets of genes. The mar phenotype is induced following
bacteria, embedded exposure to a variety of chemicals with aromatic rings such as
in a self-produced salicylates, non-antibiotic antimicrobials (triclosan) and also su-
exopolysaccharide peroxide radicals.
matrix.
Bacteria can also survive antibiotics by the formation of biofilms,
a multilayer conglomeration of diverse-species of bacteria, em-
bedded in a self-produced exopolysaccharide matrix and spe-
cially attached to surfaces including objects such as prostheses or
catheters. Antibiotic-resistance of bacteria in many infections
(e.g., gingivitis, cystic fibrosis) is attributed to the biofilms
formed by them. The precise mechanism of this resistance is still
not very clear. Bacteria detached from the biofilms are found to
be antibiotic-sensitive. Slow diffusion of antibiotics and a slow
rate of growth and metabolism are believed to contribute signifi-
cantly to antibiotic-tolerance in biofilms. Further, biofilms could
be an ideal niche for horizontal gene mobilization, allowing
resistance genes to be transferred and expressed rapidly. Conju-
gation promotes formation of biofilms in E. coli. The ability of
bacteria to respond to quorum sensing [16] is a pre-requisite for
the formation of biofilm. It is not surprising therefore that some
quorum sensing inhibitors (baicalin hydrate, cinnamaldehyde,
hamamelitannin) have recently been found to enhance the effi-
cacy of some antibiotics against pathogens.

8.1 The Antibiotic Paradox

In 1992, Professor Stuart B Levy, Director of the Center for

Adaptation Genetics and Drug Resistance at Tufts University
School of Medicine (USA), proposed the theory of Antibiotic
Paradox. It states that the widespread use of antibiotics itself is
promoting the emergence of resistant strains. The basis of this
theory lies in the fact that our body (as well as our surroundings)

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is populated by millions of bacteria most of which are susceptible Antibiotics help to

to antibiotics. The frequency of occurrence of antibiotic-resistant select resistant
bacteria is usually negligible in these naturally occurring popula- bacteria in the soil
tions. Their growth is restrained by the susceptible bacteria, by killing or
which outnumber them. Nowadays we resort to indiscriminate suppressing the
use of antibiotics on trifle illness (a pill for every ill) and even in susceptible
cases like common cold for which the antibacterial antibiotics are bacteria.
of no use. These antimicrobials annihilate or suppress the friendly
bacteria and thus provide opportunity to the antibiotic-resistant
bacteria to grow and predominate in the population. We also
often dump unused antibiotic formulations on the soil. Thus
antibiotics help to select resistant bacteria in the soil by killing or
suppressing the susceptible bacteria.

Evidences available in the literature speak volumes in support of

the theory. Following the Second World War, outbreaks of dys-
entery caused by sulfonamide-resistant Shigella sp. was a major
concern in Japan. The situation was improved by the introduction
of a number of newly discovered antibiotics viz., streptomycin,
and chloramphenicol, into the clinical practice in the year 1950.
However by 1956, again a strain of Shigella flexneri, resistant to
sulfonamides and all these three new antibiotics, was detected. In
the year 1969, the frequency of occurrence of the multidrug-
resistant strains was found to be 69% among the 10462 strains
tested. Fluroquinolones were used only in 1.4% of the cases
during 1983–85. The fluoroquilone-resistance was not detected
in E coli tested during the period, 1983 to 1990. With a sharp rise
in the use of this drug in clinical practice during 1991 to 1993,
28% of the E. coli strains tested were found to be fluoroquinolone-
resistant. A couple of years back, a team of investigators led by
Ranadhir Chakraborty, at the Department of Biotechnology in the
North Bengal University, demonstrated a correlation between the
prevalence of resistance to specific antibiotics in some river
isolates, obtained from north Bengal, and over the counter sale of
the same antibiotics in and around the town of Shiliguri.

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8.2 Other Examples

The use of antibiotics in poultries and animal farms for clinical

management of the infections of the livestock and also as a
growth promoter is well-documented. Substantial portion of the
antibiotics applied to the animals is excreted into the soil, leading
to the selection of antibiotic-resistant strains. Washed by rain and
carried to the ponds and rivers, these antibiotics enter the body of
fishes, which serve as a reservoir of resistant bacteria. Emergence
of resistant strains in fishes is also favored by the antibiotics,
used for the control of infections in fish farming. Isolation of
antibiotic-resistant bacteria from fishes and other marine organ-
isms has been reported from several laboratories across the globe.

9. Prudent Use Not a Permanent Solution

It may appear from Prof Levy’s theory that the problem of

antibiotic-resistance could be bypassed by avoiding the use of
antibiotics. In the UK a joint committee on the use of antibiotics
in Animal Husbandry and Veterinary Medicine was set up in
1968 under the Chairmanship of Prof. M M Swan. Antibiotics
were routinely used during that time in the country for the
purpose of promoting growth of animals. The committee recom-
mended a stricture on the use of therapeutically useful antibiotics
for this purpose. Use of tetracycline as a growth promoter was
prohibited in March 1971. However, during the 16 months of
prohibition the incidence of pigs carrying tetracycline-resistant
organisms was not found to decrease. Following the fall of
socialism in Eastern Europe, and also because of commercial
blockade inflicted on the country, Cubans had less access to
antibiotics during the 1990s. After 10 years, it was found that the
degree of antibiotic-resistance in the harmless bacteria isolated
from the mouth of Cubans was almost equal to that of their
counterparts from Mexico, where antibiotics is available without
prescription and also used in agriculture. Widespread resistance
to a number of therapeutically useful antibiotics was observed by
the investigators at the Centre for Cellular and Molecular Biology

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(Hyderabad, India) among bacterial isolates, obtained from dif-

ferent types of samples (soil, ice, cyanobacterial mat), collected
from Antarctica, where exposure of the organisms to antibiotics
is highly improbable. During an exchange program conducted at
the Leibniz Institute of Freshwater Ecology and Inland Fisheries
(Stechlin, Germany), it was found that incidence of chloram-
phenicol-resistance (19.69%) was not substantially lower than
that of erythromycin-resistance (24.24%) in various types of
lake-isolates, sampled from some least populated areas of north
Germany. It is well-known that use of chloramphenicol for thera-
peutic purpose was banned in the Western countries quite some
time back while erythromycin is still being used. Many other
reports of this sort reveal that prudent use of antibiotics might
help slow down the emergence of resistant strains but the strategy
cannot ensure complete reversal and disappearance of resistance.

10. Co-occurrence of Antibiotic Resistance with Tolerance

to Other Antimicrobials

Antibiotic resistance genes are often located on plasmids, which

also carry genes conferring resistance to many other chemicals
including heavy metals and disinfectants. Substances like heavy
metals and disinfectants are persistently present in the environ-
ment since they are part and parcel of the industrial civilization.
Consequently, they select resistant strains among the naturally
occurring population of bacteria and in the process, cross-select
antibiotic-resistant strains. In some cases, bacteria are found to
harbor plasmids conferring resistance to a number of antibiotics
and heavy metals. It is also known that in the absence of antibiot-
ics, resistance plasmids and their bacterial hosts co-evolve in
such a way that, after several generations, they grow better than a
strain that lacks the plasmid. The metabolic burden needed to
maintain such a big plasmid does not seem to pose any challenge
to the organisms and they are found to carry it through genera-
tions without any selection pressure. That is why it is said that
antibiotic resistance genes are easy to get but difficult to lose.

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11. Search for New Antibiotics

An antibiotic research reported by the London School of Eco-

nomics and Political Science (LSE) showed that bacterial and
parasitic diseases are the second leading cause of death world-
wide. Because of the emergence of drug-resistant ‘superbugs’,
(like MRSA and VRE), traditional antibiotics and its derivatives
are becoming nonfunctional. The whole world is thus confronted
with a looming drug crisis because of frequent emergence of
resistant strain and development of new antibiotics is desperately
needed. The search for new antibiotics during the past few
decades was mostly based on the chemical modification of the
existing drugs but in the post genomic era, molecular genetics is
being used to identify essential new targets with the help of high
throughput screening and combinatorial chemistry. Structural
biology is being applied to rapidly explore and optimize the
interactions between lead compounds and their biological targets.
Genomic tools are helping us to select for antibacterial targets and
understand bacterial resistance. On the other hand, the availabil-
ity of a plethora of natural products and combinatorial synthetic
small molecule libraries, provides ample opportunities to dig into
these resources in search of new chemotherapeutic agents. A
number of approaches to the discovery of new antibacterials are
on the anvil. Some of them are based on the forward chemical
genetic approach while the others are based on the reverse
chemical genetic strategy (Box 1).

12. Doomsday Ahead?

In the past few decades, because of the emergence of resistant

strains, therapeutic benefit of some of the potent antibiotics was
completely lost. Over nearly 20 years, from the early 1980s to the
late 1990s, not a single truly new antibiotic was introduced into
clinical use. Even now, barely a trickle has reached the market
since 1999. Only five new antibiotics were approved from 2003
to 2007, compared to the 16 approved during 1983 to 1987. The
situation for the treatment of Gram-negative infections is even
bleaker. The impermeable nature of the Gram-negative envelope

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Box 1. Chemical Genetics

Chemical genetics is a chemistry-based approach, which involves diverse small-molecule compounds to

elucidate biological pathways in a manner analogous to the mutagenesis strategies used in classical
genetics. Screening small-molecule libraries to induce a phenotype of interest represents the forward
chemical genetic approach, whereas the reverse approach involves small molecules targeting a single
protein. The prerequisite for this analysis is a collection of potential compounds (natural products,
combinatorial libraries, inventories and chemi-informatics) and high throughput functional assays (in
vitro binding assays, cellular assays, yeast, bacteria, animal models, bioinformatics).

In forward genetics, small

molecules are used to at-
tain a specific phenotype
and try to find the gene that
controls it; in reverse ge-
netics, small molecules are
used to manipulate genes
or proteins to characterize
their role by identifying the
resulting phenotype.

and presence of multiple efflux pumps in combination with other

resistance mechanisms contribute to the difficulty of this task. If
the emergence of antibiotic resistance continues to follow the
present trend, it is reasonable to assume that resistance to the new
antibiotics will emerge sooner or later. In fact, reduced suscepti-
bility to linezolid and daptomycin has already been encountered
in the clinical setting. Despite prudent usage, the life span of a
new antibiotic is cut short by the emerging resistance. So it is
evident that a continuous supply of structurally novel antibacte- If the emergence of
rial agents with multiple mode of action is needed to combat the antibiotic resistance
problem of drug resistance. The optimism associated with antimi- continues to follow the
crobial peptides because of their broad-spectrum activity, ab- present trend, it is
sence of cross-resistance with the existing antibiotics and low reasonable to assume
probability of developing resistance is hindered by their poor that resistance to the
bioavailability and high cost involved in their manufacture. Phy- new antibiotics will
sicians are often taking resort to multiple antibiotic therapies, emerge sooner or
which are likely to be more effective than using a single later.

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antibiotic. Scientists are also looking for alternative strategies

e.g., enhancing the susceptibility of bacteria to antibiotics by
using plant-derived antimicrobials (viz., thymol, carvacrol,
cinnamaldehyde, allyl isothiocyanate). Let us hope for a better
future while being aware of the precarious situation that is
prevailing at present.

13. Conclusion

The foregoing discussion on antibiotic-resistance in bacteria is

focused on the clinical impact of the phenomenon. But there are
many other aspects which are overshadowed by its devastating
effects on the prospect of chemotherapy. The history of antibiot-
ics as therapeutic agents is less than 100 years old while antibiotic
biosynthetic pathways and antibiotic-resistance genes are be-
lieved to have evolved thousands of years ago. So in nature both
antibiotic biosynthetic pathways and mechanisms involved in
antibiotic resistance must have some other significance, which
warrants extensive investigations. The multiplicity and non-speci-
ficity of efflux pumps and occurrence of resistance-conferring
genes in non-pathogenic bacteria hint at some other role of
antibiotics in evolution. It is also believed that bacteria sense
antibiotics as an environmental stress. Hence there might be some
correlation between antibiotic-resistance and stress tolerance of
bacteria. Antibiotic-resistance has been detected in many bacte-
ria isolated from extreme environments. Enhanced rate of hori-
zontal gene transfer was observed by some investigators in some
food-borne bacteria treated with sublethal levels of stress factors.
Therefore, it is obvious that the clinical aspect of antibiotic-
resistance is only the tip of the iceberg and most of the aspects in
the study in antibiotic-resistance of bacteria remain unexplored
till now. We hope that thorough investigations will provide more
insights in the years to come.

Suggested Reading

[1] C F Amabile-Cuevas, New antibiotics and new resistance, American

Scientist, Vol.91, pp.138–149, 2003.

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