BIOLOGY

PROJECT
 INDEX
Introduction
History & discovery
Usage
How do antibiotics work?
Resistance
7 types of antibiotics
5 basic mechanisms of antibiotics against
bacterial cell
Pros of taking antibiotics
Side effects of antibiotics
Experiment
Aim
Materials Required
Procedure
Observations
Conclusion
Bibliography
 INTRODUCTION
An antibiotic is a type of antimicrobial substance
active against bacteria. It is the most important
type of antibacterial agent for fighting bacterial
infection, and antibiotic medications are widely
used in the treatment and prevention of such
infections. They may either kill or inhibit the
growth of bacteria. A limited number of
antibiotics also possess antiprotozoal
activity. Antibiotics are not effective
against viruses such as the ones which cause
the common cold or influenza drugs which inhibit
growth of viruses are termed antiviral drugs or
antivirals rather than antibiotics. They are also not
effective against fungi; drugs which inhibit growth
of fungi are called antifungal drugs.
 HISTORY & DICOVERY

Prior to the beginning of the 20th Century,

infectious diseases accounted for high morbidity
and mortality worldwide. The average life
expectancy at birth was 47 years (46 and 48 years
for men and women respectively) even in the
industrialized world. Infectious diseases such as
smallpox, cholera, diphtheria, pneumonia, typhoid
fever, plaque, tuberculosis, typhus, syphilis, etc.
were rampant.

The discovery of penicillin in 1928 by Sir

Alexander Fleming (1881-1955) marked the
beginning of the antibiotic revolution. Ernst Chain
and Howard Florey purified the first penicillin,
penicillin G, in 1942 but became widely available
outside the Allied military in 1945. This marked
the beginning of the antibiotic era. This antibiotic
era witnessed the discovery of many new
antibiotics, and the period between the 1950s and
1970s was named the golden era of discovery of
novel antibiotics, and no new classes of antibiotics
have been discovered since then. After that, the
approach to discovery of new drugs was the
modification of existing antibiotics.

The antibiotic era revolutionized the treatment of

infectious diseases worldwide, although with much
success in developed countries. In the US for
example, the leading causes of death changed from
communicable diseases to non-communicable
diseases (cardiovascular disease, cancer, and
stroke), the average life expectancy at birth rose to
78.8 years, and older population changed from 4%
to 13% of the entire US population. And infectious
diseases now become the problem of elderly,
cancer patients, transplant patients, surgical
patients, patients on immune-suppressive drugs
and other at-risk groups in developed
countries. Although the developing countries also
recorded a lot of improvement in the morbidity
and mortality rate, infectious diseases still
disproportionately affect all age group in these
parts of the world. This is due to a combination of
other factors like poverty, inadequate public health
measures, poor sanitation, poor vaccination
coverage, etc.
 USAGE

Antibiotics are used to treat or prevent bacterial

infections, and sometimes protozoan infections.
When an infection is suspected of being
responsible for an illness but the responsible
pathogen has not been identified, an empiric
therapy is adopted. This involves the
administration of a broad spectrum
antibiotic based on the signs and symptoms
presented and is initiated pending laboratory
results that can take several days.
When the responsible pathogenic microorganism
is already known or has been identified, definitive
therapy can be started. This will usually involve
the use of a narrow-spectrum antibiotic. The
choice of antibiotic given will also be based on its
cost. Identification is critically important as it can
reduce the cost and toxicity of the antibiotic
therapy and also reduce the possibility of the
emergence of antimicrobial resistance. To avoid
surgery, antibiotics may be given for non-
complicated acute appendicitis.
Antibiotics may be given as a preventive
measure and this is usually limited to at-risk
populations such as those with a weakened
immune system (particularly in HIV cases to
prevent pneumonia), those
taking immunosuppressive drugs, cancer patients,
and those having surgery. Their use in surgical
procedures is to help prevent infection
of incisions. They have an important role in dental
antibiotic prophylaxis where their use may
prevent bacteraemia and consequent infective
endocarditis. Antibiotics are also used to prevent
infection in cases of neutropenia particularly
cancer-related.
The use of antibiotics for secondary prevention of
coronary heart disease is not supported by
current scientific evidence, and may actually
increase cardiovascular mortality, all-cause
mortality and the occurrence of stroke.
 HOW DO ANTIBIOTICS WORK?

Antibiotics work by blocking vital processes in

bacteria. They kill the bacteria or stop it from
spreading. This helps the body’s natural immune
system to fight the infection. There are many
types of antibiotics. They work against different
types of bacteria.
 Some antibiotics affect a wide range of
bacteria. They are called broad spectrum
antibiotics (eg, amoxicillin and gentamicin).
 Some antibiotics affect only a few types of
bacteria. They are narrow spectrum
antibiotics (eg, penicillin).
There are many types of antibiotics. Not all of
them work in the same way. For instance,
penicillin destroys bacterial cell walls. Other
antibiotics can affect the way the cell works.
Doctors choose an antibiotic based on the bacteria
that often cause a certain infection. Sometimes
your doctor will do a test. This is to identify the
exact type of bacteria causing your infection. It
also checks how it reacts to certain antibiotics.
 RESISTANCE

A significant threat to the achievements of the

antibiotic era is the antibiotic resistance, which is
the ability of the bacteria to resist the effect of
antibiotic for which they were initially sensitive
to. Resistance results from natural resistance in
certain types of bacteria (present even before
antibiotics were discovered), genetic mutations in
microbes, by one species acquiring resistance
from another and selection pressure from
antibiotic use that provides a competitive
advantage for mutated strains. Suboptimal
antibiotic doses, especially from misuse of
antibiotics, help in the stepwise selection of
resistance. Examples of significant resistant
pathogens in the world are Penicillin-Resistant
Streptococcus pneumonia (PRSP), Methicillin-
Resistant Staphylococcus aureus (MRSA),
Vancomycin-Resistant Enterococci (VRE) and
Multiple-Drug-Resistant Gram-Negative Bacilli
(MDRGNB).

Resistant organisms are difficult to treat,

requiring higher doses or alternative drugs, which
are possibly more toxic and expensive. According
to the Centres for Diseases Control and
Prevention (CDC), at least 2 million people
become infected with antibiotic-resistant bacteria
and more than 23,000 people die annually as a
consequence of these infections. This is expected
to be more in developing countries where
communicable diseases remain the leading causes
of death. To worsen the situations are the
emerging and re-emerging infectious diseases,
lack of development of new classes of antibiotics
and continuing increasing antibiotic resistance,
including superbugs (bacteria with accumulated
resistance to almost all available antibiotics), at
alarming and dangerous levels worldwide.

Natural antibiotic resistance in bacteria may

predate the discovery of some antibiotics, and it
may be difficult to control since this is one of the
natural coping mechanisms for their survival.
However, the majority of the acquired antibiotics
resistances are promoted by the socioeconomic
and behavioural factors, especially in the
developing countries. Some of these factors
include misuse of antibiotics by health
professionals, misuse and inappropriate
antibiotics use by unskilled practitioners and
laypersons. Other factors are poor quality
medicines, dissemination of resistant organisms
through overcrowding and unhygienic condition,
inadequate hospital infection control practices,
inadequate surveillance, poverty, lack of
resources to implement strategies against
antibiotic resistance and lack of political will.
 7 TYPES OF ANTIBIOTICS

Although there are well over 100 antibiotics, the

majority come from only a few types of drugs.
These are the main classes of antibiotics.
1.Penicillins such as penicillin and amoxicillin
2.Cephalosporins such as cephalexin (Keflex)
3.Macrolides such as erythromycin (E-
Mycin), clarithromycin (Biaxin),
and azithromycin (Zithromax)
4.Fluoroquinolones such as ciprofolxacin
(Cipro), levofloxacin (Levaquin),
and ofloxacin (Floxin)
5.Sulphonamides such as co-trimoxazole
(Bactrim) and trimethoprim (Proloprim)
6.Tetracyclines such as tetracycline (Sumycin,
Panmycin) and doxycycline (Vibramycin)
7. Aminoglycosides such
as gentamicin (Garamycin)
and tobramycin (Tobrex)
 Common Antibacterial Drugs by Mode of Action

Mode of Action Target Drug Class

Inhibit cell wall Penicillin-binding β-lactams: penicillins,

biosynthesis proteins cephalosporins,
monobactams,
carbapenems

Peptidoglycan Glycopeptides
subunits

Peptidoglycan Bacitracin
subunit transport

Inhibit biosynthesis of 30S ribosomal Aminoglycosides,

proteins subunit tetracyclines

50S ribosomal Macrolides,

subunit lincosamides,
chloramphenicol,
oxazolidinones

Disrupt membranes Lipopolysaccharide, Polymyxin B, colistin,

inner and outer daptomycin
membranes

RNA Rifamycin
Inhibit nucleic acid
synthesis
DNA Fluoroquinolones

Folic acid synthesis Sulfonamides,

enzyme trimethoprim
Antimetabolites
Mycolic acid Isonicotinic acid
synthesis enzyme hydrazide

Mycobacterial ATP Diarylquinoline

Mycobacterial adenosine synthase
triphosphate (ATP)
synthase inhibitor
 5 BASIC MECHANISMS OF
ANTIBIOTICS AGAINST
BACTERIAL CELLS
PROS OF TAKING ANTIBIOTICS
 Antibiotics can slow the growth of and kill
many types of infection.
 In some cases, such as before surgery,
antibiotics can prevent infection from
occurring.
 Antibiotics are fast-acting; some will begin
working within a few hours.
 They are easy to take: Most antibiotics are
oral medications. Your doctor may decide to
give you an injection, if it is imperative that
the medicine gets into your system quickly.

SIDE EFFECTS OF ANTIBIOTICS

Antibiotics are screened for any negative effects
before their approval for clinical use, and are
usually considered safe and well tolerated.
However, some antibiotics have been associated
with a wide extent of adverse side effects ranging
from mild to very severe depending on the type of
antibiotic used, the microbes targeted, and the
individual patient. Side effects may reflect the
pharmacological or toxicological properties of the
antibiotic or may involve hypersensitivity
or allergic reactions. Adverse effects range from
fever and nausea to major allergic reactions,
including photodermatitis and anaphylaxis
Common side effects of oral antibiotics
include diarrhoea, resulting from disruption of the
species composition in the intestinal flora,
resulting, for example, in overgrowth of
pathogenic bacteria, such as clostridium
defficile. Taking probiotics during the course of
antibiotic treatment can help prevent antibiotic-
associated diarrhoea. Antibacterials can also
affect the vaginal flora, and may lead to
overgrowth of yeast species of the
genus candida in the vulvo-vaginal
area. Additional side effects can result
from interaction with other drugs, such as the
possibility of tendon damage from the
administration of a quinolone antibiotic with a
systemic corticosteroid.
Some antibiotics may also damage
the mitochondria, a bacteria-derived organelle
found in eukaryotic, including human,
cells. Mitochondrial damage cause oxidative
stress in cells and has been suggested as a
mechanism for side effects
from fluroquinolones. They are also known to
affect chloroplasts

EXPERIMENT
AIM: To study the effect of antibiotics on
microorganisms
Materials required: Potato, agar, dextrose,
beef, peptone, NaCl, Sodium Bicarbonate,
distilled water, five different types of
antibiotics(such as penicillin, streptomycin,
aureomycin, Terramycin, and
chloromycetin), syringe, oven sterilised
petridishes, flasks,, beakers, pipette, garden
soil, glass marker pen, etc.
EXPERIMENTAL PROCEDURE :
A. Preparation of Culture Medium
1. Potato Dextrose Agar Medium.
Take 200g of peeled potato chips. Boil them
in 500 ml of water in a beaker for 15
minutes. Squeeze the potato pul thus
obtained through a muslin cloth and keep it
in a flask. Take 20g of agar in a beaker and
warm it with 500ml of water. Mix both the
solutions of potato and agar and add 20g of
dextrose to it. Thus one litre of PDA
medium is prepared.
Autoclave the medium with 15 pounds
pressure for 15 minutes.

2. Meat Extract Agar Medium.

Weigh 3g beef extract, 10g peptone, 5g NaCl
and mix these in 1 litre of distilled water.
Heat he mixture to 65`C stirring until the
materials are completely dissolved. Filter
the mixture through filter paper and adjust
the pH to 7.2 to 7.6 by adding a bit of
sodium bicarbonate. Now, add 20g agar to
the broth and autoclave at 15 pounds
pressure for 15 minutes.

B. Effect of Antibiotics on Soil

Microorganisms
 (i) Take 2g of soil and dissolve it in
10ml of water in a beaker.
Let the soil particle settle down.
(ii) Take 6 oven sterilised petridishes
and pour 1ml of suspension in each
of the plates. Now pour on ml of each
antibiotics separately in 5
petridishes with the help of syringe,
and mark them with marker pen.
Leave the 6th petridish without
antibiotic to serve as control.
(iii) Pour PDA or meat extract agar in
each of the petridishes and mix the
suspension by rotating the
petridishes. Leave the petridishes
undisturbed at a warm place for a
week.

OBSERVATION:
Colonies of microorganisms appear on the
culture medium, count the colonies in each
petridish and present your observations in
the following table:
S.no. Antibiotics No. of colonies
in a petridish
1. Penicillin ……
2. Streptomycin ……
3. Terramycin ……
4. Chloromycetin ……
5. Control ……
6.

The effect of different antibiotics on the

microorganisms can be assessed by
counting the number and size of the
colonies growing in the petridishes.

PRECAUTIONS:
1. Do not expose the culture of the
petridish to the atmosphere.
2. Sterilise the dishes properly in the
oven before use.
3. Use proper kind of stains for different
types of microorganisms.
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