ANAOMY

BIO 0GP
CotiaNO

EVOLUTION
STUDY OF ANTIBIOTICS
ON MICROBS

OLÁSSXH
BÌODOGY
INVESTIGATORY
PROJECT
 ACKNOWLEDGEMENT

In the accomplishment of this project successfully, many people have best

owned upon me their blessings and the heart pledged support, this
tmeI utlising to thank all the people who have been concerned
m

with this wholesome project:

Primardy, myself would thank God for
being able to conplete this project with success. Then, I would ike to
thank or Principal maam,Sagufta Parvin and biology ma'an,
Subhashree Mishra, whose valuable guidance has been the ones that
helped me patch this project and make ta Full- PROOF SUcCESS. I
would despaurately thank my parenty for their valuable suggestions and
superintendence which served as theMAJOR CONTRIBUTOR towards the
completion of the project.
Lastly, thanks to all my class mates who had helped ne a lot.
 CERTIFICATE
This is to certify that of Class 12th B'of Mother'y
Publie School of roll no. has successfully completed his/her Biology
project file. He/She hay taken care with utmost sincerity in completion of
hisher project. AU the work related to the project is done by the
candidate himselfherself. The approach towards the subject has been
profownd and scientific.

I that this project is up-to

certify my expectations& and
as per the guidelines issued by the CBSE.
PRINCIPAL MA'AM

INTERNAL EXAMINER EXTERNAL EXAMINER

 EFFECTSOF
ANTIBIOTICS ON
MICROORGANISMS
 INDEX
Introduction

History and Discovery

Goals
How do antibiotics work?
Why are antibiotics important?
Resistance:

Antibiotic Resistance

How is Resistance spread?

7 Types of antibiotics
5 Basic mechanism of antibiotic action

against bacterial cell

Pros of taking antibiotics

Side effects of taking antibiotics
Experimernt:
Aim

>Materials Required
Procedure
>Observations
>Conclusion

Bibliography
 1

INTRODUCTION

Antibiotics are chemicals that kill or inhibit the

growth of bacteria and ae used to treat bacterial
They are produced in nature by soil
infections.

bacteria and fungi. This gives the microbe an

advantage when competing forfood and water and
other limited resources in a particular habitat, as
the antibiotic kils off their competition. Only
substances that target bacteria are called
antibiotics.

•Antiseptics are used to sterilize surfaces of living

tissue when the risk of infection is high, such as
during surgery.

Disinfectants are non-selective antimicrobial,

killing a wide range of micro-organisms including
bacteria. They are used on non-living surfaces,for
example in hospitals.
 2

HISTORY AND DISCOVERY

In 1928, at St. Mary Hospital, London, Alexander

Fleming discovered Penicillin. Fleming was, a bit

disorderly in his work and accidently discovered

penicillin. Upon returning from holiday in Suffolk in

1928, he noticed that a fungus, Penicillium notatum,

had contaminated a culture plate of Staphylococcus
bacteria he had accidently left uncovered.

The discovery of penicillin changed the world of

medicine enormously. With its development,
infections that were previously severe and often fatal,

like bacterial endocarditis, bacterial meningitis and

pneumococcal pneumonia, could be easily treated.

Antibiotics were widely used for treating soldiers

during World War Il, caring for battle wound

infections and pneumonia. By the mid-to-late 1940s,
it became widely accessible for the general public.
 GOLS

1. To study the effect of antibiotics on

micro-organisms(bacteria).
2. To understand its mechanism.
 HOW D0 ANTIBIOTICS
WORK?
Antibiotics are used to treat bacterial infections.

Antibiotics take advantage of the difference between

the structure of bacterial cell and the host's cell.

They either prevent the bacterial cells from

multiplying so that the bacterial population remains
the same, allowing the host's defense mechanism to
fight the infection or kill the bacteria, for example
stopping the mechanism responsible for building
their cell walls.

An antibiotic can also be classified according to the

range of pathogens against which it is effective.

PenicillinG will destroy only a few species of bacteria

and is known as narrow spectrum bacteria.

Tetracycline is effective against a wide range of

organisms and is known as broad spectrum bacteria.
WHY ARE ANTIBIOTICS
IMPORTANT?

The introduction of antibiotics into medicine

revolutionized the way infectious diseases were
treated. Between 1945 and 1972, average human life

expectancy jumped by eight years, with antibiotics

used to treat infections that were previously likely to

kill patients. Today antibiotics are one of the most
common classesof drugs used in medicine and make
possible many of the complex surgeries that have
become routine around the world.

If we ran out of effective antibiotics, modern

medicine would be set back by decades. Relatively

minor surgeries, such as appendectomies, could

become life threatening, as they were before
antibiotics became widely available. Antibiotics are
sometimes used in a limited numbers of patients
 6

before surgery to ensure that patients do not contact

any infections from bacteria entering open cuts.
Without this precaution, the risk of blood poisoning
would become much higher, and many of the more
complex surgeries doctors now perform may not be
possible.
 RESISTANCE
1.Antibiotic resistance

Bacteria are termed drug-resistantwhen they are no

longer inhibited by an antibiotic to which they were
previously sensitive. The emergence and spread of
antibacterial-resistant bacteria has continued to grow
due to both the over-use and misuse of antibiotics.

Treating a patient with antibiotics causes the

microbes to adapt or die; this is known as 'selective

pressure'. If a strain of a bacteria species acquires

resistance to an antibiotic, it will survive the
treatment. As the bacterial cell with acquired
resistance multiplies, this resistance is passed on to
its offspring. In ideal conditions some bacterial cells

can divide every 20 minutes;therefore after only 8

hours in excess of 16 million bacterial cells carrying
resistanceto that antibiotic could exist.
2. How is resistance spread?

Antibiotic resistance can either be inherent or

acquired. Some bacteria are naturally resistance

to some antibiotics due to their physiological

characteristics. This is inherent resistance.

Acquired resistance occurs when a bacterium

that was originally sensitive to an antibiotic

develops resistance. For example, resistance

genes can be transferred from one plasmid to
another plasmid or chromosome, or resistance
can occur due to a random spontaneous
chromosomal mutation.
 9

7TYPES OFANTIBIOTICS

Although there are well over 100 antibiotics, the

majority come from only a few types of drugs. These
are the main classesof antibiotics.

ANTIBIOTIC EXAMPLE
PENICILLIN amoxicillin

CEPHALOSPORINS cephalexin
MACROLIDES erythromycin
FLUOROQUINOLONES ofloxacin

SULPHONAMIDES bactrim
TETRACYCLINES tetracycline
AMINOGLYCOSIDES gentamicin
 10

FIVE BASIC MECHANISMS OF

ANTIBIOTIC ACTION
AGAINST BACTERIAL CELLS:

inhibition of cell wall synthesis (mostcommon

mechanism).
inhibition of protein synthesis (translation).
Alteration of cel membranes.
IInhibition of Nucleic acid synthesis.
IAntimetaboliteactivity.

The ribosome is a major target for Antibiotics.

Drugs inhibit ribosome function either by
interfering in messenger RNA translation or by
blocking the formation of peptide bonds at the
peptidyl transferase centre. These effectsare the

consequence ofthe binding of drugs to ribosomal

subunits.
 11

PROS OF TAKING
ANTIBIOTICS:

Antibiotics can slow down the growth of bacteria

and kill many types of infections.

• 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.
 12

SIDE EFFECTS OF
TAKING
ANTIBIOTICS:

The most common side effects of antibiotics is- it

affects the digestive system. Antibiotics commonly

cause the following side effects:

•Diarrhea
•Nausea
•Vomiting
•Rash
•Upset stomach
• Sensitivity to sunlight,when taking tetracycline
•With certain antibiotics or prolonged use, fungal
infections of the mouth, digestivetract, and
vagina.
EXPERIMENT
AIM: To see the effect of antibiotics on bacteria

count.

MATERIALS REQUIRED:
•10 test-tubes of sterilized water

•10 PCA(Agar) plates

•Bunsen burner

•Graduated cylinder
•Ethanol (Used for sterilizing; just flame is enough
most cases)
•Glass hockey stick

•Pipettes
•Refrigerator
•Incubator (A warm cabinet for growing bacteria)
•Scale
•Large beaker
•Hot plate

Sample antibiotic
 14

PROCEDURE:
Step 1:

Prepare a culture media plate for growing bacteria.

Step 2:

Get a sample of polluted water for test. Mix 2 ml of

polluted water with 10 ml chicken broth in a test tube
and incubate it for 24 hours so the bacteria will

reproduce and increase. Usually this is done on a

device that constantly moves, so the bacteria can
freely move in the liquid. Most likely you will not have
a vibrator, so it is good if you shake the test tube a
few times during this incubation period.

Step: 3
While the bacteria are being incubated, prepare
some antibiotic disks as described here. (Antibiotic

disks can also be purchased from biology suppliers.)

Break an antibiotic capsule (here using Ampicillin)

and empty the contents in a clean petri-dish. One
capsule will be enough for hundreds of disks.
 15

Dispose of the plastic shell and add a few drops of

water to the remaining powder. Cut some filter

papers in small pieces and soak them in the antibiotic

solution. Let the disks dry in a clean space. You may
cover them with another filter paper to protect them
from dust.

Although they are known as antibiotic disks, you can

cut them in small squares.
 16

The reason that we use filter paper, is that other

papers often have starch and other polymers that
may affect the results of our experiments. Filter

paper is pure cellulose fiber.

Step 4:

Use the bacteria that you have grown in step 2 and

prepare dilution of bacteria.

1.Prepare 1:10 dilution of thesample. To do this, take

2ml of the sampleand blend it with 18mlof distilled
water.

2.Pipette 0.1ml of each dilution onto a Plates Count

Agar (PCA) plate.

3.Take a glass hockey stick submersed in ethanol and

run it through a flame to sterilize it. (Glass hockey
 17

stick is a glass rod bent on one end like a hockey

stick. It is used to spread bacteria on the surface of
agar plate. You may use a steelspoon instead.)
4.Let it cool and use it to spread dilution around the
plate.

5.Do this on two plates for each of the five different

dilutions.

6.Place an antibiotic disk on the plate of dilution.

7.Incubate the plate at 35 degrees Celsius for 24
hours and then count the bacterial colonies.

8.Take 3 nutrient agar plate and added 0.5ml of the

solution on each of plates.

9.Leaving one plate without any antibiotics, placing

one antibiotic disk on the second plate and two

antibiotic disks on the third plate. All plates were
incubated for 48 hours.
 18

OBSERVATIONS:

PCA1

PCA2

PCA3
 19

CONCLUSION:

The growth of bacteria around the antibiotic disks

is less.Inhibition zones are more in the plates with
more antibiotic disks.

Hence, antibiotics stop the proliferation of

bacteria.
 20

BIBLIOGRAPHY:

wwW.emedicinehealth.com

microbiologysociety.org

www.scienceproject.com

medicalnewstoday.com

www.scribd.com

www.studocu.com