An antibiotic is an agent that either kills or inhibits the growth of a

microorganism. The term antibiotic was first used in 1942 by Selman

Waksman and his collaborators in journal articles to describe any substance
produced by a microorganism that is antagonistic to the growth of other
microorganisms in high dilution.

This definition excluded substances that kill bacteria but that are not produced
by microorganisms (such as gastric juices and hydrogen peroxide). It also
excluded synthetic antibacterial compounds such as sulfonamides.

Many antibacterial compounds are relatively small molecules with a molecular

weight of less than 2000 atomic mass units.

With advances in medicinal chemistry, most modern antibacterial are semi-

synthetic modifications of various natural compounds. These include, for
example, beta-lactam antibiotics, which include penicillin (produced by fungi
in the genus Penicillium), cephalosporin, and carbapenems. Compounds that
are still isolated from living organisms are aminoglycosides, whereas other
antibacterial—for example, sulfonamides, quinolones, and oxazolidinones—
are produced solely by chemical synthesis.

Following this, many antibacterial compounds are classified based on

chemical/biosynthetic origin into natural, semi-synthetic, and synthetic.
Another classification system is based on biological activity; in this
classification, antibacterial are divided into two broad groups according to
their biological effect on microorganisms:

✓ Bactericidal agents that kill bacteria

✓ Bacteriostatic agents slow down or stall bacterial growth.

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Antibiotic resistance is a form of drug resistance whereby some (or, less
commonly, all) sub-populations of a microorganism, usually a bacterial
species, can survive after exposure to one or more antibiotics; pathogens
resistant to multiple antibiotics are considered multidrug resistant (MDR) or,
more colloquially, superbugs.

Antibiotic resistance is a serious and growing phenomenon in contemporary

medicine and has emerged as one of the pre-eminent public health concerns
of the 21st century, as it pertains to pathogenic organisms (the term is
especially relevant to organisms that cause disease in humans). A World
Health Organization report released April 30, 2014, states, "this serious threat
is no longer a prediction for the future, it is happening right now in every
region of the world and has the potential to affect anyone, of any age, in any
country. Antibiotic resistance–when bacteria change so antibiotics no longer
work in people who need them to treat infections–is now a major threat to
public health."
In the simplest cases, drug-resistant organisms
may have acquired resistance to first-line
antibiotics, thereby necessitating the use of
second-line agents. Typically, a first-line agent
is selected based on several factors including
safety, availability, and cost; a second-line
agent is usually broader in spectrum, has a less
favorable risk-benefit profile, and is more
expensive or, in dire circumstances, may be
locally unavailable. In the case of some MDR
pathogens, resistance to second- and even third-line antibiotics is, thus,
sequentially acquired, a case quintessentially illustrated by Staphylococcus
aureus in some nosocomial settings. Some pathogens, such as Pseudomonas
aeruginosa, also possess a high level of intrinsic resistance.

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It may take the form of a spontaneous or induced genetic mutation, or the
acquisition of resistance genes from other bacterial species by horizontal gene
transfer via conjugation, transduction, or transformation. Many antibiotic
resistance genes reside on transmissible plasmids, facilitating their transfer.
Exposure to an antibiotic naturally selects for the survival of the organisms
with the genes for resistance. In this way, a gene for antibiotic resistance may
readily spread through an ecosystem of bacteria. Antibiotic-resistance
plasmids frequently contain genes conferring resistance to several different
antibiotics. This is not the case for Mycobacterium tuberculosis, the bacteria
that causes Tuberculosis, since evidence is lacking for whether these bacteria
have plasmids. Also M. tuberculosis lack the opportunity to interact with other
bacteria in order to share plasmids.

Genes for resistance to antibiotics, like the antibiotics themselves, are ancient.
However, the increasing prevalence of antibiotic-resistant bacterial infections
seen in clinical practice stems from antibiotic use both within human medicine
and veterinary medicine. Any use of antibiotics can increase selective
pressure in a population of bacteria to allow the resistant bacteria to thrive
and the susceptible bacteria to die off. As resistance towards antibiotics
becomes more common, a greater need for alternative treatments arises.
However, despite a push for new antibiotic therapies, there has been a
continued decline in the number of newly approved drugs. Antibiotic
resistance therefore poses a significant problem.
The growing prevalence and incidence of infections due to MDR pathogens is
epitomized by the increasing number of familiar acronyms used to describe
the causative agent and sometimes the infection; of these, MRSA is probably
the most well-known, but others including VISA (vancomycin-intermediate S.
aureus), VRSA (vancomycin-resistant S. aureus), ESBL (Extended spectrum
beta-lactamase), VRE (Vancomycin-resistant Enterococcus) and MRAB
(Multidrug-resistant A. baumannii) are prominent examples. Nosocomial
infections overwhelmingly dominate cases where MDR pathogens are
implicated, but multidrug-resistant infections are also becoming increasingly
common in the community.

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Although there were low levels of preexisting antibiotic-resistant bacteria
before the widespread use of antibiotics, evolutionary pressure from their use
has played a role in the development of multidrug-resistant varieties and the
spread of resistance between bacterial species. In medicine, the major
problem of the emergence of resistant bacteria is due to misuse and overuse
of antibiotics. In some countries, antibiotics are sold over the counter without
a prescription, which also leads to the creation of resistant strains. Other
practices contributing to resistance include antibiotic use in livestock feed to
promote faster growth.] Household use of antibacterial in soaps and other
products, although not clearly contributing to resistance, is also discouraged
(as not being effective at infection control). Unsound practices in the
pharmaceutical manufacturing industry can also contribute towards the
likelihood of creating antibiotic-resistant strains. The procedures and clinical
practice during the period of drug treatment are frequently flawed — usually
no steps are taken to isolate the patient to prevent re-infection or infection by
a new pathogen, negating the goal of complete destruction by the end of the
course (see Healthcare-associated infections and Infection control).

Certain antibiotic classes are highly associated with colonization with

"superbugs" compared to other antibiotic classes. A superbug, also called
multiresistant, is a bacterium that carries several resistance genes. The risk
for colonization increases if there is a lack of susceptibility (resistance) of the
superbugs to the antibiotic used and high tissue penetration, as well as broad-
spectrum activity against "good bacteria". In the case of MRSA, increased
rates of MRSA infections are seen with glycopeptides, cephalosporins, and
especially quinolones. In the case of colonization with Clostridium difficile, the
high-risk antibiotics include cephalosporins and in particular quinolones and
clindamycin.

Of antibiotics used in the United States in 1997, half were used in humans and
half in animals; in 2013, 80% were used in animals.

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Antibiotic resistance is becoming more and more common. Antibiotics and
antimicrobial agents are drugs or chemicals that are used to kill or hinder the
growth of bacteria, viruses, and other microbes. Due to the prevalent use of
antibiotics, resistant strains of bacteria are becoming much more difficult to
treat. These "super bugs" represent a threat to public health since they are
resistant to most used antibiotics. Current antibiotics work by disrupting so-
called cell viability processes.

Disruption of cell membrane assembly or DNA translation are common modes

of operation for current generation antibiotics. Bacteria are adapting to these
antibiotics making them ineffective means for treating these types of infection.
For example, Staphylococcus aureus have developed a single DNA mutation
that alters the organism's cell wall. This gives them the ability to withstand
antibiotic cell disruption processes. Antibiotic resistant Streptococcus
pneumoniae produce a protein called MurM, which counteracts the effects of
antibiotics by helping to rebuild the bacterial cell wall.

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Researchers are attempting to develop new types of antibiotics that will be
effective against resistant strains. These new antibiotics would target the
bacteria's ability to become virulent and infect the host cell. Researchers at
Brandeis University have discovered that bacteria have protein "switches" that
when activated, turn "ordinary" bacteria into pathogenic organisms. These
switches are unique in bacteria and are not present in humans. Since the
switch is a short-lived protein, elucidating its structure and function was
particularly difficult. Using nuclear magnetic resonance (NMR) spectroscopy,
the researchers were able to regenerate the protein for one-and one-half
days. By extending the time frame that the protein was in its "active state," the
researchers were able to map out its structure. The discovery of these
"switches" has provided a new target for the development of antibiotics which
focus on disrupting the activation of the protein switches.

Monash University researchers have demonstrated that bacteria contain a

protein complex called Translocation and Assembly Module (TAM). TAM is
responsible for exporting disease causing molecules from the inside of the
bacterial cell to the outer cell membrane surface. TAM has been discovered in
several antibiotic resistant bacteria. The development of new drugs to target
the protein would inhibit infection without killing the bacteria. The researchers
contend that keeping the bacteria alive, but harmless, would prevent the
development of antibiotic resistance to the new drugs.

Researchers from the NYU School of Medicine are seeking to combat

antibiotic resistance by making resistant bacteria more vulnerable to current
antibiotics. They discovered that bacteria produce hydrogen sulfide to counter
the effects of antibiotics. Antibiotics cause bacteria to undergo oxidative
stress, which has toxic effects on the microbes. The study revealed that
bacteria produce hydrogen sulfide to protect themselves against oxidative
stress and antibiotics. The development of new drugs to target bacterial gas

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defenses could lead to the reversal of antibiotic resistance in pathogens such
as Staphylococcus and E. coli.

These studies indicate how highly adaptable bacteria are in relation to the
application of antimicrobial treatments. Antibiotic-resistant bacteria have
become a problem not only in hospitals, but in the food industry as well. Drug-
resistant microbes in medical facilities lead to patient infections that are more
costly and difficult to treat. Resistant bacteria in turkey and other meat
products have caused serious public health safety issues. Some bacteria may
develop resistance to a single antibiotic agent or even multiple antibiotic
agents. Some have even become so resistant that they are immune to all
current antibiotics. Understanding how bacteria gain this resistance is key to
the development of improved methods for treating antibiotic resistance.

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1. Sterilized Petri dishes
2. Sterilized culture tubes with media
3. Transfer loops
4. Forceps
5. Flask
6. Beaker
7. Burner
8. Penicillin
9. Aureomycin
10. Hay
11. Alcohol
12. Agar
13. Starch
14. Distilled water

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1. To 200ml of distilled water in a flask, I added 8 grams of agar powder and 2
grams of starch. Then putting a few pieces of dry hay into the medium I covered
the flask with an Inverted beaker. Boiling the medium for 5 minutes and then
cooling the medium to room temperature. After that placing the flask in a warm
place. Within 2-3 days, formation of scum of cloudy suspension appeared on
the medium indicating the growth of Bacillus subtilis.

2. Taking culture tubes with agar medium and heating the test tubes in warm
water to melt agar. Cooling each test tube so that I can hold it in my hand and
the agar remains liquid. After that removing the cotton plug and I passed the
mouth of the test tube through the burner flame twice. Flaming the transfer loop
after dipping it in alcohol and I let it cool. After that picking up a loop full of
bacterial culture from flask and then I transferred it to the warm agar in the
culture tube. Flaming the loop and the mouth of the culture tube and then I
replaced the cotton plug. Rolling the culture tube of warm agar between palms
to I mixed the bacteria well with agar.
*Transferring the bacteria should be done as quickly as possible.

3. After that I took sterilized Petri dishes. Removing the cotton plug and flamed
the mouth of the culture tube. Then I lifted the cover of the Petri dish at an angle
45 Degree and then quickly pouring the medium of the culture tube into the
bottom half the dish. Removing the culture tube and replacing the cover tube
into the bottom half of the dish. Removing the culture tube and replace the cover
of the Petri dish. Moving the covered Petri dish along the tabletop to distribute
the medium evenly. Then I allowed the agar to cool. After that I prepared two
Petri dishes and marked them A & B.

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4. I prepared Penicillin and Aureomycin solution by dissolving the powdered
drugs in distilled water. Then I cut down a few discs of filter paper of 1 cm
diameter. Then I soaked a disc in each of the penicillin and Aureomycin
solutions. Dipping the forceps in alcohol and the I passed the forceps’ tip quickly
over the burner flame. Using the sterilized forceps, I put Penicillin and
Aureomycin soaked discs at two distant sites of Petri dish A. Considering Petri
dish B as control. Then I kept both the Petri dishes undistributed in warm place
to allow the bacteria to grow. Then I observed the Petri dishes for several days.

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The area around the antibiotic discs in the Petri dishes will be clear. In other
areas, colonies of bacteria will be observed. Then after examining the clear
area in each Petri dishes for few more days. A few very colonies may appear
in the clear areas. These are the colonies of resistant strains of the bacteria.

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Antibiotic drugs killed most of the bacterial strain, hence the areas appeared
clear. However, a few strains which were resistant in the bacterial population
survived and produced colonies later. This proves the resistant strain to
antibiotics were present in the bacterial population.

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1. Comprehensive Laboratory Manual In Biology-XII
2. Biology Text For Class XII – NCERT
3. https://www.1000sciencefairprojects.com/Biology/drug-resistance.php

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