Advances in Animal and Veterinary Sciences.

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Review Article
An Overview of Mechanisms and Emergence of Antimicrobials Drug
Resistance
Sujeet Kumar*, Bhoj Raj Singh
Indian Veterinary Research Institute, Izatnagar, Bareilly, India
*Corresponding author: [email protected]

ARTICLE HISTORY ABSTRACT

Received: 2013–09–16 Emergence of antimicrobial drug resistance in bacteria is widely explored but still ill understood.
Revised: 2013–10–24 Antibiotic resistance is a complex phenomenon due to multiple ways of its acquisition, mechanism
Accepted: 2013–10–24 of action and spread. Antibiotic resistance may be innate, present in microbes due to their inherent
inbuilt metabolic and structural components. Other type of innate resistance is one which has
been detected in strains isolated even before the era of the use of any antibiotics and often specific
Key Words: ADR, MDR, with bacterium. The most important and problematic is the acquired drug resistance. It spread
XDR, TDR, Superbug, rapidly with the use of antimicrobials in one or other realm of ecosystem. Inactivation of
Mechanism, Emergence antimicrobials occurs by some common mechanism such as drug inactivation/degradation by
bacterial enzymes or alteration in the bacterial targets or expulsion of drug out of the bacterial
cells (efflux) or by preventing the entry of drug into bacterium. Here some important aspects of
mechanism of antimicrobial drug resistance and its emergence are discussed.

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ARTICLE CITATION: Kumar S and Singh BR (2013). An overview of mechanisms and emergence of antimicrobials drug resistance.
Adv. Anim. Vet. Sci. 1 (2S): 7 – 14.

INTRODUCTION designated as nonprescription use.

The field of medical and veterinary sciences was revolutionized According to recent estimates, 80% of total antibiotics
by the discovery of antibacterial drug by the middle of the 20th produced in world are used in animals. In USA total human
century. It resulted to dramatic improvement in human and antibiotic use was estimated at 8 billion pounds per annum
animal health and productivity. The diseases like tuberculosis, while in animals it amounts to record 29.9 billion pounds per
bubonic plague, cholera which were the scourge throughout the year (Marshal and Levy, 2011). During 2006, China used 97,000
human civilization came under complete control. Soon, tons of antibiotics in animal feed out of total 210,000 tons of
scientists and planner entered in euphoria that bacterial production. Today China produces almost 400,000 tons of
diseases will be just a history in very short time (Singh, 2011, antibiotics annually and most of the production is meant for
2013). However, euphoria remained short–lived, as bacteria animal husbandry (Li et al., 2013a). Most of the antibiotics used
responded against the antibiotics by manifesting different in human or animals as therapeutic drugs have been used as
forms of resistance. The situation is rapidly turning now in the antibiotic growth promoters (AGPs) in one or other kind of
favor of bacterial pathogens. The number of new antibacterial animal farming. Evidences suggest the role of AGPs in
agents has started to dwindle. But bacteria are evolving day emergence of multiple drug resistance (Singh, 2011). In view of
over the day more and more clever mechanisms of resistance, this, Sweden first took the initiative to prohibit the use of
and posing a serious threats to human and animal health (Levy antimicrobials in animal feed. European Union and USA
and Marshall, 2004; Singh, 2013). restricted or banned several antimicrobials for non–
Use of Antimicrobials in Animals prescription use in animals and aquaculture. The commonly
In animals antibiotics are used for the following purposes used antimicrobials used as AGPs include, amoxicillin,
(Schwarz et al., 2001): ampicillin (AGPs in aquaculture), ardacin (bovine AGP, a
 Therapeutic uses: To treat infections and applied in glycopeptides withdrawn from EU in 1997), avilamycin (AGP
therapeutic doses (higher than used for preventive, control for broilers withdrawn from EU), avoparcin (AGP, a
and production purposes) and for a short period. glycopeptides withdrawn from EU in 1997), bacitracin (AGP
 Preventive or control uses: For prevention of a disease, to for poultry, swine, and beef cattle, withdrawn from EU in 1999
control the spread of one or other disease in a situation but available in USA), bambermycin (AGP for poultry, swine
where disease is likely to occur due to environmental and cattle, withdrawn from EU in 2006 but available in USA),
conditions and exposure of disease causing organisms. colistin (in broiler, swine and cattle feed in Japan), efrotomycin
 Production or growth promotion uses: It is to (AGP for swine), erythromycin (AGP for poultry, cattle and
increase daily body weight gain and/ or to enhance feed swine), lasalocid (AGP for cattle, poultry, sheep and rabbits
efficiency by administering doses lower than those required to approved in EU and USA), lincomycin (AGP for chicken and
treat or prevent disease over an extended period of time. Often swine, approved in USA), monensin (AGP in bovine and
poultry, withdrawn from EU for bovine use but authorized as
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Kumar et al (2013). Mechanisms and Emergence of Antimicrobials Drug Resistance
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Advances in Animal and Veterinary Sciences. 1 (2S): 7 – 14
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poultry coccidiostat), neomycin (AGP for swine and poultry increase in international trade of animals and animal products
approved in USA), narasin (poultry feed coccidiostat, AGP for is also a concern in the spread of antibiotic resistance. (Tiwari
cattle approved in USA), nourseothricin (swine AGP et al., 2013a; WHO, 2013). To overcome the hurdles of
withdrawn from EU), olaquindox (swine AGP, withdrawn due antibiotics resistance various alternative emerging novel
to toxicity in EU and Canada, available in USA), oleandomycin therapies are coming into picture such as herbal medication,
(poultry and swine AGP), ormetoprim (poultry AGP for ethno–veterinary medicines, bacteriophage therapy, cytokine
prevention of fowl cholera and other infections), pristinamycin therapy, mycophage therapy, panchgavya therapies etc. which
(AGP for wide use), procaine penicillin (AGP in poultry and are opening new avenues to fight against these superbugs
swine, withdrawn in USA), roxarsone (AGP for poultry and (Dhama et al., 2013a,b; Tiwari et al., 2013a,b).
swine), salinomycin (swine AGP), spiramycin (swine AGP Antibiotics and their Spectrum
withdrawn from EU in 1999), sulfonamides (aquaculture, The term antibiotic was coined by Selman Waksman, a Nobel–
swine, chicken AGP), tetracycline (AGP for poultry, swine and laureate soil microbiologist who discovered streptomycin.
cattle withdrawn from EU, authorized in USA), tiamulin Antibiotic is defined as a compound produced by a microbe
(swine AGP), tylosin (swine AGP, withdrawn from EU, which kill or inhibit the growth of another microbe. Hundreds
available in USA) and virginiamycin (AGP for broilers, of antibiotic molecules of natural, semi–synthetic and synthetic
withdrawn from EU and available in USA) (Samanidou and nature are used for treating infections. The mechanism of action
Evaggelopoulou, 2008). by these antimicrobial compounds is summarized in Table 1.
Excessive and unnecessary use of antimicrobials, The different antimicrobials used in veterinary medicine and
particularly as growth promoters in livestock and poultry may their mechanism of action has been has been extensively
lead to a great risk to human health (WHO, 2013). In 1998, the reviewed by Schwarz and Chaslus–Dancla (2001) and Andre
European Union banned the use of antimicrobials for growth and Bryskier (2005).
promotion in animals as per WHO recommendations. An

Table 1: An overview of antimicrobial action by different class of antibiotics

Mechanism of action Examples
1. Penicillins (amoxicillin, ampicillin, cloxacillin, carbenicillin, all
penicillins)
2. Cephalosporins (cefotaxime, ceftriaxzone, cefapime, cefoxitin,
Interfering in bacterial cell wall synthesis
cefoperazone)
3. Carbapenems (imipenem, ertapenem, meropenem)
4. Monobactams (aztreonam)
5. Glycopeptide (vancomycin)
1. Chloramphenicol
Inhibiting protein synthesis through binding to 50S subunit 2. Macrolides (erythromycin, azithromycin)
of ribosome 3. Ketolide (telithromycin)
4. Lincosamide (clindamycin, lincomycin)
1. Aminoglycosides (streptomycin, gentamicin, amikacin, neomycin,
Inhibiting of protein synthesis through binding with 30S kanamycin)
subunit of ribosome 2. Tetracyclines (oxytetracycline, methacyclin, doxycyclin,
tigecycline)
Fluoroquinolones (ciprofloxacin, enerofloxacin, norfloxacin,
Inhibiting DNA synthesis
ofloxacin)
Inhibiting RNA synthesis Rifampin
Inhibiting dihydrofolate reductase activity Sulfonamides, trimethoprim
Disrupting bacterial membrane Polymyxins (polymyxin B, polymyxin E or colistin

History of Antibiotic Resistance among the bacterial pathogens by conjugation. Many other
Sulfonamide was introduced in 1937 as a first effective mechanisms like transformation and transduction were also
antimicrobial agent. Since then the development of resistance observed with respect to antibiotics resistance. In the light of
has plagued their therapeutic use. Penicillin was discovered by many new findings, the area has been the matter of recent
renowned microbiologist Alexander Fleming in 1928. But before review in many leading journals (Alekshun and Levy, 2007;
its introduction as a therapeutic agent the enzyme (bacterial Allen et al., 2010; Davies and Davies, 2010). In brief, the
penicillinase/ β–lactamases) which destroy it was identified sequence of antibiotics discovered and emergence of resistance
(Abraham and Chen, 1940). Later methicillin was developed by is briefed in Table 2.
chemically modifying the penicillin to prevent cleavage by β– Superbugs
lactamases. But just within 3 years of introduction of the The term “superbugs” refers to microbes having high level of
methicillin, the drug resistant bacteria were reported (Davies drug resistance and causing enhanced morbidity and mortality
and Davies, 2010). In 1944, streptomycin was introduced for the to no response to therapeutic efforts. Super–bug infections limit
treatment of tuberculosis. Now tuberculosis treatment is facing the therapeutic options for the clinicians; extend the periods of
the serious challenge due to development of resistance against hospitalization and thus the treatment cost. In some cases,
streptomycin and most of other antimicrobials used in super–resistant strains have also acquired increased virulence
tuberculosis kit. The whole concept of drug resistance started and enhanced transmissibility. Over the years, continued
changing by as early as mid–1950. It became evident that selective pressure by different antimicrobials led to multidrug
antibiotic resistance is genetically transferable. Later, it was resistance (MDR) in human and animal pathogens. Some of the
observed that antibiotic resistance genes could be disseminated most problematic MDR organisms include methicillin–resistant

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Staphylococcus aureus (MRSA), extensively drug–resistant (XDR) (ESBL), Pseudomonas aeruginosa, Escherichia coli and Acinetobacter
Mycobacterium tuberculosis, vancomycin–resistant Staphylococcus baumannii (Foster, 2004; Arias and Murray, 2009; Davies and
aureus (VRSA), vancomycin–resistant enterococci (VRE), Davies, 2010).
Klebsiella pneumoniae bearing extended–spectrum β–lactamases

Table 2: Timeline for development of antibiotics and antibiotic–drug resistance

Decade Entry of antibiotics in market Antibiotics resistance detected
Emergence of vancomycin–resistant staphylococcal infections
2001 to Introduction of broader spectrum
spread of extended–spectrum β–lactamases (ESBL) and metallo– β –
2010 fluoroquinolones, telithromycin, tigecycline
lactamases (MBL) among Gram negative bacteria.
1991 to Introduction of oral extended spectrum Emergence of vancomycin–resistant enterococci, emergence of
2000 cephalosporin multi–drug resistant Mycobacterium tuberculosis.
Introduction of cefotaxime, clavulanic acid,
1981 to
amoxicillin, imipenem–cilastatin, norfloxacin, Spread of methicillin–resistant Staphylococcus infections.
1990
aztreonam
1971 to Introduction of carbenicillin, cefoxitin, cefaclor Ampicillin resistant infections become frequent. Increasing trend of
1980 opportunistic pathogen infection.
1961 to Introduction of gentamicin, ampicillin, Emergence of gentamicin resistant Pseudomonas, emergence of
1970 cephalothin, amikacin methicillin–resistant staphylococcal infections.
1951 to Introduction of erythromycin, vancomycin and
Penicillin–resistant infections become clinically significant.
1960 methicillin
1941 to Introduction of streptomycin, chloramphenicol Penicillin became available to the public and its widespread use in
1950 and chlortetracycline animals started by 1950.
1930 to Efficacy of penicillin in humans shown, sulfonamides introduced in
Introduction of sulfonamide
1940 animal feed
Before Resistance to several antibiotics in strains of Murray collection from
Discovery of penicillin (1929)
1930 pre–antibiotic era.

Tuberculosis infects nearly one–third of the world Clostridium difficile is a Gram–positive, spore forming and
population. Emergence of MDR strain of M. tuberculosis is toxin–producing anaerobe. It causes severe intestinal infections
seriously compromising the TB therapy (Blanchard, 1996). In as a result of extensive use of antibiotics. The depletion of
the last decade, extremely drug–resistant (XDR) strains intestinal microflora by antibiotics and subsequent
resistant to four or more front–line antibiotics used against colonization by C. difficile is thought as primary cause of
tuberculosis. The XDR strains are now being reported from antibiotic associated diarrhea (AAD) or post–antibiotic
several parts of the world and posing severe threat to diarrhea (Abigail et al., 2011).
community health. Moreover, the emergence of total drug Antimicrobial Resistance Mechanism
resistant (TDR) strains is compelling to think for the coming The mechanism of antibiotics resistance has extensively been
back to pre–antibiotics era (Alanis, 2005; Davies and Davies, reviewed (Hawkey, 1998; Alanis, 2005; Higgins, 2007; Alekshun
2010). and Levy, 2007; Arias and Murray, 2009; Davies and Davies,
The Gram–negative pathogens such as Escherichia coli and 2010). The mechanisms of resistance for different class of
Klebsiella pneumoniae cause variety of diseases in humans and antimicrobials have been summarized in Table 3. Several
animals. Antibiotic resistance has been observed in these different mechanisms may work together to confer resistance to
pathogens against lactam class of antibiotics. These pathogens a single antimicrobial agent. Overall resistance mechanisms
show a great deal of diversity and produces upto 1000 different adopted by bacteria are classified (Brötz–Oesterhelt and
types of β–lactamases (Hancock, 1998). Brunner, 2008) as:
Pseudomonas aeruginosa has emerged as a major nosocomial  Reduction in entry or access to the target site of
threat apart from being involved in burn wound infection. It has antimicrobial drug
evolved resistance against β–lactam and aminoglycosides group  Activation of efflux mechanism to expel antimicrobial
of antibiotics (Livermore, 2002; Breidenstein et al., 2011). agents from the bacterial cell
Acinetobacter baumannii is another Gram–negative pathogen  Enzymatic degradation or modification of antimicrobials
associated with nosocomial infection. It got equipped with all either inside or outside of the bacterial cell
different kind of resistance genes due to their remarkably high  Modification of antimicrobial drug target(s) within the
ability of natural transformation (Poirel and Nordmann, 2006; bacterial cell
Arias and Murray, 2009). Reduction in Entry or Access to the Target Site of
Staphylococcus aureus is regarded as one of the most Antimicrobial Drug
notorious superbugs (Foster, 2004; Lindsay and Holden, 2004). Antimicrobial compounds require entry and or access into the
The pathogen resides as nasal commensal in almost 30% of bacterial cell to interfere with the normal function of bacteria.
human population. It is now recognized as a major nosocomial Porin channels serve as passage for antibiotics to cross the
infection. Methicillin was the first anti–resistance antibiotic bacterial outer membrane. Few bacteria protect themselves by
developed against the penicillinases. But within three years of prohibiting entry of antimicrobials into their cell (Mah and
introduction of methicillin, a methicillin–resistant S. aureus O'Toole, 2001), viz., many Gram–negative bacteria reduce the
(MRSA) emerged. Due to development of MDR strain of S. uptake of aminoglycosides and beta lactam by modifying the
aureus, the acronym now denotes multidrug–resistant S. aureus. cell membrane porin channel numbers, size, and selectivity.
This prevents aminoglycosides and beta lactams to reach their

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intended targets, ribosome and the penicillin–binding proteins Enzymatic Degradation or Modification of Antimicrobials
(PBPs), respectively. This strategy has been observed in Either Inside or Outside of the Bacterial Cell
Pseudomonas aeruginosa and Klebsiella spp. against imipenem (a Degradation mechanism of antibiotics resistance (Wright,
beta–lactam antibiotic), and vancomycin intermediate resistant 2005) is the first ever antibiotic resistance mechanism observed
S. aureus (VISA) strains have thickened cell wall to trap the drug shortly after the discovery of penicillin, even before its
before entry in to the cell (Howden et al., 2010) commercial application (Abraham and Chain, 1940). In the
Activation of Efflux Mechanism to Expel Antimicrobial process bacteria destroy the active component of the
Agents from the Bacterial Cell antimicrobial agent. It is the most common mechanism to
Efflux mechanism of antibiotic resistance has extensively been inactivate β–lactam group of antibiotic, aminoglycoside and
reviewed (Poole, 2005; Piddock, 2006; Higgins, 2007). Bacteria chloramphenicol. A classic example is the hydrolytic
possess membrane pumps to move lipophilic or amphipathic degradation of the β–lactam ring in penicillin and
molecules across the cell wall. Efflux pump is true variant of cephalosporin by the bacterial β–lactamases. The inactivation
these existing pumps. Antibiotic producing bacteria widely use product, penicilloic acid, is ineffective in binding to penicillin
such pump to move antibiotics out of the cells as fast as they are binding proteins (PBPs), thereby can’t prevent cell wall
produced. Therefore, it constitutes a protective mechanism to synthesis. Besides, the strategy has also been used by both
prevent bacteria getting killed by their own chemical weapons. Gram positive and Gram negative bacteria to inactivate
Several pathogenic bacteria possess trans–membrane proteins aminoglycosides and chloramphenicol through acetylation,
which act as an efflux pump. This pumps the antibiotic out of adenylation and phosphorylation (Davies, 1994; Jana and Deb,
the cell as fast as it enters. This results in low intracellular 2006).
concentrations of antibiotics insufficient to elicit antibacterial Modification of Antimicrobial Target Within the Bacterial
effect. The efflux pumps are sometime selectively specific to Cell
extrude antibiotics like tetracycline, macrolides, lincosamide, Modification in target sites allows some bacteria to avoid
and streptogramins. Many other pumps possess ability to expel recognition by antimicrobial agents (Spratt, 1994). Such
multiple drugs which can expel a variety of structurally strategy has been observed in methicillin resistant S. aureus
different antimicrobials (Piddock, 2006). This strategy has been (MRSA) through change or acquisition of different PBPs, in
observed in E. coli and other Enterobacteriaceae against vancomycin resistant Enterococcus (Walsh and Howe, 2002), in
tetracycline and chloramphenicol, in staphylococci against sttreptomycin resistant Mycobacterium (through modification of
macrolides and streptogramins and in S. aureus and Streptococcus ribosomal proteins or 16s rRNA), mutations in RNA polymerase
pneumoniae against fluoroquinolones (Li et al., 2013a). lead to rifampicin resistance in M. tuberculosis (Rattan et al.,
1998; Rodríguez–Verdugo et al., 2013) and mutations in DNA
gyrase lead to resistance for quinolones in many Gram–negative
bacteria, S. aureus and Streptococcus pneumonia (Hooper, 2001)

Table 3: Mechanism of resistance to different classes of antimicrobials

Antimicrobial Mechanism of Specific means to
Examples
class Resistance achieve resistance
Destruction of lactam ring by β−lactamases Resistance to penicillins, cephalosporins,
Degradation of
making it incapable to bind with penicillin and aztreonam in Enterobacteriaceae and
antibiotic
binding protein (PBPs). Micrococaceae family.
Beta–lactams Methicillin and oxacillin resistant
Altered target Mutational changes PBPs.
staphylococci.
Imipenem resistance of K. pneumonia,
Decreased uptake Decreased porin channel formation. Enterobacter aerogenes, and Pseudomonas
aeruginosa.
Glycopeptides Alteration in cell wall precursor so
Altered target Vancomycin resistant enterococci.
decreases the binding of vancomycin
Enzymatic Aminoglycoside resistance in G+ve and
Acetylation
modification G–ve bacteria.
Aminoglycosides Change in number or character of porin Aminoglycoside resistance in G–ve
Decreased uptake
channels. bacteria.
Modification of ribosomal proteins or 16s
Altered target Streptomycin resistant Mycobacterium spp.
rRNA.
Decreased uptake Alterations in the outer membrane
Quinolone resistant G–ve bacteria and
and increased diminishes uptake of drug. Activation of
Quinolones staphylococci.
excretion efflux pumps to excrete.
Fluoroquinolone resistant G+ve and G–ve
Altered target Changes in DNA gyrase.
bacteria.

Molecular Mechanism of Antimicrobial Resistance resistance are all genetically encoded. These genetic
The molecular mechanism of antibiotics resistance has been mechanisms are classified into two types;
extensively reviewed time to time (Blanchard, 1996; Hawkey, 1. Intrinsic resistance
1998; Walsh, 2000; Alanis, 2005; Alekshun and Levy, 2007). The 2. Acquired resistance
bacterial abilities to adopt various strategies for antibiotic 1. Intrinsic resistance:

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It is the innate ability of bacteria to resist antimicrobial effect of 2. Acquired resistance:

particular antibiotic class through its inherent structural or It is the ability of bacteria to resist the activity of a particular
functional characteristics. Such resistance can also be called as antimicrobial agent to which it was earlier susceptible. This is
“insensitivity” as those microbes have never been susceptible to mediated by mutation or horizontal gene transfer
that particular drug. The classical example is resistance of (transformation, transduction or conjugation) which brings
anaerobes to aminoglycosides and of G–ve bacteria to changes in bacterial genome. This brings alteration in the
vancomycin (Fajardo et al., 2008). Different type of intrinsic bacterial structural and functional characteristics leading to
resistance existing in different class of antibiotics is presented resistance against a particular antibiotic. The genetic
in Table 4. The natural insensitivity can be due to lack of drug mechanism for acquiring resistance by different antimicrobial
targets, inability of drug to enter bacterial cell, expulsion of class has been summarized in Table 5.
antimicrobials by chromosomally encoded efflux pump and
innate production of antibiotic inactivating enzymes.

Table 4: Intrinsic resistance in bacteria

Organisms Antibiotics Mechanism
Gram–positive Lack of penicillin binding proteins which can effectively
Aztreonam (beta–lactam)
bacteria bind aztreonam
Gram–negative Large molecule of vancomycin is unable to penetrate outer
Vancomycin
bacteria membrane of G–ve bacteria
β−lactamases produced by the bacteria destroy ampicillin
Klebsiella spp. Ampicillin
before it reaches the PBP targets
Stenotrophomonas β−lactamase produced by the bacteria destroy imipenem
Imipenem
maltophilia before it bind with PBP target.
Lactobacillus and
Vancomycin Unable to bind with cell wall precursor
Leuconostoc
Sulfonamides, trimethoprim, tetracycline, In–effective intracellular concentrations of antibiotics due
Pseudomonas aeruginosa
chloramphenicol to lack of uptake
Limited uptake of aminoglycosides by protein of electron
Aminoglycosides
transport chain
Enterococcus spp.
β−lactam antibiotics like penicillin,
Lack of penicillin binding proteins
cephalosporins and monobactam

Table 5: Mechanism of acquired resistance in bacteria

Mechanism Antibiotics Mechanism
Point mutation in rpoB gene of Mycobacterium tuberculosis coding for β–subunit of RNA
Rifampicin
polymerase, the binding site for rifampicin.
Mutations Fluoroquinolones Mutation in the quinolone resistance determining region (QRDR) of GyrA and ParC/GrlA
Mutations in the chromosomal gene specifying dihydrofolate reductase in E. coli,
Trimethoprim
Haemophilus influenzae
Acquisition of mecA genes by MRSA. The penicillin binding proteins (PBPs) insensitive to
Methicillin
Horizontal gene β–lactam antibiotics inhibition are produced.
transfer Sulphonamide Acquisition of foreign folP gene.
Vancomycin Acquisition of vanA and vanB gene by enterococci.

Mutation posses only a single copy of rRNA genes in place multiple

Mutation is defined as “spontaneous change in DNA sequence copies by fast–growing bacteria like E. coli. So, it makes easier
within the gene”. A change within a single nucleotide base pair for mycobacteria to gain mutational alteration in 16S rRNA or
brings corresponding change in one or more amino acids. This ribosomal protein S12 for resistance against streptomycin.
consequently changes the affinity of antimicrobials towards the Mycobacterium tuberculosis acquires isoniazid resistance through
targeted site. mutational alteration in katG gene (encoding catalase), inhA
In bacteria, mutations naturally occur due to errors in gene (target for isoniazid) oxyR gene and aphC gene (Hazbón et
DNA polymerase activity, insertions, deletions, and al., 2006; Laurenzo and Mousa, 2011).
duplications. Due to DNA polymerase error, bacteria have a
Hydrolytic inactivation of lactam ring by β–lactamases is
spontaneous mutation rate of about 0.0033 mutations in each
the most common resistance mechanism in penicillin and
DNA replication cycle. However, the mutation rate varies
cephalosporin group of antibiotics. The parental β–lactamase
between the genes.
gene appears to have originated from environmental microbes.
Mutational alteration in the antibiotic target sites is the
reason behind the development of multidrug–resistant (MDR) However, bacteria acquired the resistance to newer β–lactam
mycobacterial infections. Streptomycin resistance in antibiotics by a series of point mutations within the lactamase
gene. Such mutations are common in members of
Mycobacterium tuberculosis occurs more frequently than in E. coli.
Streptomycin works through its binding to 30S sub–unit of Enterobacteriaceae. Penicillin binding protein (PBP) is
ribosome. Therefore, its activity is reduced once mutational responsible for binding with β−lactam antibiotics and
alteration happens in ribosomal protein S12 of 16S ribosomal inhibition of cell wall synthesis. A mutational change in the
RNA (rRNA). The slow growing bacteria like M. tuberculosis

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mecA gene brings an alternative PBP (PBP2a) which in turn resistance genes present in pathogenic bacteria have origin from
leads for methicillin resistance in S. aureus. antibiotic producing soil microbes. The quantification of
Horizontal Gene Transfer antimicrobial resistance genes in antibiotic–producing bacterial
Genetic elements like plasmid, transposon and integrons carry strains reflects that a significant number of these strains are
the antibiotic resistance genes (Boucher et al., 2007). These naturally multidrug resistant (D’Costa et al., 2006). Further,
elements act as vectors and transfer resistance genes to other environmental microbes have been evaluated for their capacity
bacteria belonging to the members of the same species, or to to use antibiotics as a sole carbon and nitrogen source. Many of
another species or even a different genus (Dzidic and the bacterial strains belonging to proteobacteria, Burkholderia
Bedekovic, 2003). Transformation, conjugation and spp. and pseudomonads grow well on aminoglycosides,
transduction are the three different mechanisms in bacteria for fluoroquinolones, and other classes of antimicrobials (Dantas et
horizontal gene transfer (HGT). The mechanism of HGT al., 2008). Streptomycetes produce a variety of β–lactamases. This
transfer in various antibiotics group has been summarized in is supposed to be the source of β–lactam resistance in clinical
Table 5. isolates (Kumarasamy et al., 2010). The environmental Kluyvera
Transformation involves the uptake of short naked DNA species have been found to be the origins of the CTX–M genes
fragments and their homologous recombination in naturally (Cantón and Coque, 2006). The quantum of data is sufficient to
competent bacteria. This is commonly observed in the species assume that the most potential pool of antibiotics resistance
of streptococci, meningococci, Acinetobacter etc. In the recent genes lies among the environmental microbes (Walsh, 2013).
years, Acinetobacter and other environmental genera have been Antibiotics Application in Agriculture
associated to transmit the resistance genes from environmental Apart from its therapeutic application in human and animals,
microbes to clinical pathogens (Riesenfeld et al., 2004). antibiotics are widely used in agriculture, aquaculture, poultry
Conjugation involves the cell to cell contact via sexual pilli industry and farm animals (Alderman and Hastings, 1998;
to transfer the piece of DNA. Sex pilli is formed by the Teuber, 2001; Wegener, 2003; Dibner and Richards, 2005;
responsible genes which is present only in the donor bacteria. Cabello, 2006; Kemper, 2008; Li et al., 2013b). All these
Ultimately, the piece of DNA fragments having the resistance facilitate a vast reserve for antibiotic resistance genes in the
genes is transferred from resistant donors to previously natural habitat. The most commonly used antibiotic in
susceptible bacteria. Recent studies have demonstrated that gut agriculture is streptomycin and oxytetracycline (McManus et
microbiota of human and animals frequently transmit the al., 2002). The excessive application of streptomycin resulted in
diverse ranges of antimicrobial resistance genes through development of resistant strains of Erwinia amylovora,
conjugation. Pseudomonas spp. and Xanthomonas campestris. Avoparcin is a
Transduction involves the transfer of DNA from one glycopeptide antibiotic. It has been extensively used as a
bacterium into another via bacterial viruses called growth promoter in poultry and pig farms. This has resulted to
bacteriophage. It is less commonly linked with the transfer of development of resistance against vancomycin, one of the drugs
antibiotic resistance genes compared to transformation and of last resort to treat human infections (Bager et al., 1997).
conjugation. However, phages are frequently associated with Therefore, it is necessary to curb the excessive use of antibiotics
the formation of mobile genetic elements encoding the to prevent the spread of antibiotic resistance.
resistance and virulence genes. Anthropogenic Activity
Comparative analysis of plasmid from pre–antibiotic to Waste disposal and wastewater treatment plants are rich
the post–antibiotic era suggests that antibiotics resistance reservoirs for antimicrobial resistance genes and resistant
genes were present even in the pre–antibiotic era (Hughes and microbes (Fick et al., 2009). This facilitates the horizontal gene
Datta, 1983). But the emergence of multi–resistance plasmids transfer from these sources to human pathogens. This demands
among the pathogens probably happened in the past five a better control of antibiotic release and environmental
decades as a pressure of antibiotics use. The integron, a mobile disposal. Recent work suggests that river water contaminated
DNA element, has two conserved segments flanking a central with quinolones enriches the qnr genes in water–borne bacteria
region carrying the antibiotic resistance gene. The integrons and might be responsible for the transfer of qnr genes to
and their site–specific recombinase (integrase) enzyme are pathogenic bacteria (Hirsch et al., 1999).
associated with the formation of multiple resistance plasmid Bacterial Housekeeping Genes
and transposon. Housing keeping genes having role in protection from
Source of Antibiotics Resistance Genes antimicrobials including antibiotics in environmental bacteria
Evolution of antibiotics resistance genes in bacterial pathogens might be the need to survive because soil, water and terrestrial
is a complex process. Growing number of evidence suggests lives produces variety of challenges. From there, the pathogenic
that environment has reserve pool of antibiotic resistance genes and commensal microbes may also acquire genes of their need.
(Martínez, 2008). Most of the antibiotic resistance genes In recent past several studies on herbal–drug resistance in
acquired through horizontal gene transfer are expected to have pathogenic bacteria have revealed that it is not only use of
originated from environmental microbes. Along with this antimicrobials which might be associated with existence/
various anthropogenic activities and evolution of existing emergence of drug resistance, microbes may have resistance to
bacterial enzymes are considered as source of resistance genes. several types of herbal drugs which might have never been used
Environmental Microbes in therapeutics (Singh et al., 2013). This phenomenon of
Role of environmental microbes in spreading antibiotics resistance to non–use antimicrobials can easily be explained on
resistance has been extensively explored (Riesenfeld et al., the basis of presence of one or other house–keeping gene. Some
2004; Allen et al., 2010). Most antibiotics used for treating of the bacterial housekeeping genes such as the sugar kinases
infection are produced by environmental microbes. Antibiotic– and acyltransferases might have evolved to modify
producing bacteria protect themselves from the lethal effect of aminoglycoside antibiotics with the use of antimicrobials too
antibiotics through modification of targets and pumping it out (Davies and Davies, 2010).
through efflux pump as soon as it is formed. Discovery of these In conclusion, it can be said that multiplicity of
pathways in Streptomycetes led to the assumption that antibiotic antimicrobials is not more than the ways to microbes know and

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Advances in Animal and Veterinary Sciences. 1 (2S): 7 – 14
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have ability to evolve to counter those and emergence of the isoniazid resistance mutations and evolution of multidrug–resistant
multiple drug resistance is the percussion. Mycobacterium tuberculosis. Antimicro. Agents Chemother. 50: 2640–2649.
Higgins CF (2007). Multiple molecular mechanisms for multidrug resistance
transporters. Nature. 446: 749–757.
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