Biology Investigatory Project

Topic: Study of Effects of

Antibiotics on Microorganisms

Prepared By Rahul Vimalkanth Roll No: 33

 Contents

Certificate of Authenticity.........................................................................................................................2
Acknowledgment........................................................................................................................................3
Objective of the Project..............................................................................................................................4
Introduction................................................................................................................................................4
Mechanisms................................................................................................................................................5
Prokaryotic bacteria....................................................................................................................................6
How antibiotic resistance spreads..............................................................................................................7
Penicillin......................................................................................................................................................9
Streptomycin.............................................................................................................................................10
Terramycin................................................................................................................................................11
Experiment................................................................................................................................................12
Requirements........................................................................................................................................12
Procedure..............................................................................................................................................13
A. Preparation of culture Medium.................................................................................................13
B. Effects of antibiotics on soil microorganisms.............................................................................13
Bibliography..............................................................................................................................................15
 Certificate of Authenticity

This is to certify that Rahul Vimalkanth, a student of class XII A Roll No. 33 has successfully completed
the research project on the topic Study of Effect of Antibiotics on Microorganisms under the guidance
of Mrs. Krishnaveni mam subject teacher in year 2022-2023. The references taken in making this project
have been declared at the end of this report.

Signature Signature Signature

(Subject Teacher) (Examiner) (Principal)
 Acknowledgment

This project would not have been feasible without the proper rigorous guidance of biology teacher Mrs.
Krishnaveni mam, who guided me throughout this project in every possible way. An investigatory
project involves various lab experiments, which have to obtain the observations and conclude the
reports on a meaningful note. Thereby, I would like to thank Mrs. Krishnaveni mam for guiding me on a
systematic basis and ensuring that in completed all my research with ease. Rigorous hard work has put
in this project to ensure that it proves to be the best. I hope that this project will prove to be a breeding
ground for the next generation of students and will guide them in every possible way.
Objective of the Project

The aim of this project is to study the effect of antibiotics on micro – organisms. To study the effect of
antibiotics on micro-organisms.

Introduction

The term "antibiotic" was coined by Selman Waksman in 1942 to describe any substance produced by a
microorganism that is antagonistic to the growth of other microorganisms in high dilution. This original
definition excluded naturally occurring substances that kill bacteria but are not produced by
microorganisms (such as gastric juice and hydrogen peroxide) and also excluded synthetic antibacterial
compounds such as the sulfonamides . Many antibiotics are relatively small molecules with a molecular
weight less than 2000 atomic mass units.

Antibacterial is an alternative name. Antibiotics belong to the broader group of antimicrobial

compounds, used to treat infections caused by microorganisms, including fungi and protozoa.

Antibiotics are the organic secretion produced by micro – organisms, which in low concentrations are
antagonistic to the growth of other micro – organisms (mostly pathogens). Antibiotics have proved very
useful in combating several bacterial diseases in man an animal. Antibiotics are commonly obtained
from actinomycetes and some eubacteria. Some of the important antibiotics are streptomycin,
Aureomycin, Terramycin, Chloromycetin, erythromycin, neomycin etc.

Soil is a natural medium that harbors several types of microorganism .These micro- organisms can be
frown on culture media. The effect of different types of antibiotics can be studied on the growth of
microorganism growing in culture medium. This is an important subject, therefore the study of effect of
antibiotics on micro - organism has been taken for the present project.

Mechanisms

Schematic representation of how antibiotic resistance evolves via natural selection. The top section
represents a population of bacteria before exposure to an antibiotic. The middle section shows the
population directly after exposure, the phase in which selection took place. The last section shows the
distribution of resistance in a new generation of bacteria. The legend indicates the resistance levels of
individuals.
Prokaryotic bacteria

DNA transfer between prokaryotic cells occurs in bacteria and archaea, although it has been mainly
studied in bacteria. In bacteria, gene transfer occurs by three processes. These are bacterial virus
(bacteriophage)-mediated transduction, plasmid-mediated conjugation, and natural transformation.
Transduction of bacterial genes by bacteriophage appears to reflect an occasional error during
intracellular assembly of virus particles, rather than an adaptation of the host bacteria. The transfer of
bacterial DNA is under the control of the bacteriophage's genes rather than bacterial genes.
How antibiotic resistance spreads
The emergence of antibiotic resistance is an evolutionary process that is based on selection for
organisms that have enhanced ability to survive doses of antibiotics that would have previously been
lethal. Antibiotics like Penicillin and Erythromycin, which used to be one-time miracle cures are now less
effective because bacteria have become more resistant. Antibiotics themselves act as a selective
pressure that allows the growth of resistant bacteria within a population and inhibits susceptible
bacteria. Antibiotic selection of pre-existing antibiotic resistant mutants within bacterial populations was
demonstrated in 1943 by the experiment. Survival of bacteria often results from an inheritable
resistance. Any antibiotic resistance may impose a biological cost. Spread of antibiotic-resistant bacteria
may be hampered by reduced fitness associated with the resistance, which is disadvantageous for
survival of the bacteria when antibiotic is not present. Additional mutations, however, may compensate
for this fitness cost and aids the survival of these bacteria.

Antibiotic-resistant microorganisms, sometimes referred to as "superbugs", may contribute to the re-

emergence of diseases which are currently well-controlled. For example, cases of tuberculosis (TB) that
are resistant to traditionally effective treatments remain a cause of great concern to health
professionals. Every year, nearly half a million new cases of multidrug-resistant tuberculosis (MDR-TB)
are estimated to occur worldwide.

Microorganism is able to survive exposure to an antibiotic. Genes can be transferred between bacteria
in a horizontal fashion by conjugation, transduction, or transformation. Thus, a gene for antibiotic
resistance which had evolved via natural selection may be shared. Evolutionary stress such as exposure
to antibiotics then selects for the antibiotic resistant trait. Many antibiotic resistance genes reside on
plasmids, facilitating their transfer. If a bacterium carries several resistance genes, it is called
multiresistant or, informally, a superbug or super bacteria.

The primary cause of antibiotic resistance is antibiotic use both within medicine and veterinary
medicine. The greater the duration of exposure the greater the risk of the development of resistance
irrespective of the severity of the need for antibiotics.
Penicillin

Penicillin’s are antibiotics used to treat bacterial infections that are derived from the antibiotic penicillin.

Penicillin antibiotics stop bacteria from multiplying by preventing bacteria from forming the walls that
surround them. The walls are necessary to protect the bacteria from their environment, and to keep the
contents of the bacterial cell together. Bacteria cannot survive without a cell wall. Penicillin antibiotics
are most effective when bacteria are actively multiplying and forming cell walls.

Today, many derivatives of penicillin have been developed that inhibit more types of bacteria than the
original life-saving drug.
Streptomycin

Streptomycin is an aminoglycoside antibiotic originally isolated from the bacteria Streptomyces griseus.
It is the first discovered aminoglycoside antibiotic. Its use is primarily treating aerobic gram-negative
bacterial infections, such as brucellosis, tularemia, plague.

Streptomycin also is used as a pesticide, to combat the growth of bacteria beyond human applications.
Streptomycin controls bacterial diseases of certain fruit, vegetables, seed, and ornamental crops. A
major use is in the control of fireblight on apple and pear trees. As in medical applications, extensive use
can be associated with the development of resistant strains. Streptomycin could potentially be used to
control cyanobacterial blooms in ornamental ponds and aquaria. While some antibacterial antibiotics
are inhibitory to certain eukaryotes, this seems not to be the case for streptomycin, especially in the
case of anti-fungal activity.
Terramycin

Terramycin (Oxytetracycline Hydrochloride and Polymyxin B Sulfate Ophthalmic Ointment) is an

antibiotic, bright yellow in color, possessing potent antimicrobial activity. It is one of the most versatile
of the broad-spectrum antibiotics and is effective in the treatment of infections due to gram-positive
and gram-negative bacteria, both aerobic and anaerobic, spirochetes, rickettsia, and certain of the larger
viruses.
Polymyxin B sulfate is one of a group of related antibiotics derived from Bacillus polymyxa. The
polymyxins are rapidly bactericidal, this action being exclusively against gram-negative bacteria. The
broad-spectrum effectiveness of Terramycin against both gram-positive and gram-negative organisms is
enhanced by the effectiveness of polymyxin B against infections associated with gram-negative
organisms, especially those due to Pseudomonas aeruginosa, where polymyxin B is the antibiotic of
choice.
Experiment

Requirements

1. Potato
2. Agar
3. Dextrose
4. Distilled water
5. Four different types of antibiotics (such as penicillin , streptomycin , aureomycin , terramycin)
6. Syringe
7. Oven sterilized petridish
8. Flasks
9. Beakers
10. Pipettes
11. Garden soil
12. Glass marker pen
Procedure

A. Preparation of culture Medium

1. Potato Dextrose Agar (PDA) Medium.

a. Take 200 g of peeled potato chips.
b. Boil them with 500 ml of water in a beaker for 15 minutes.
c. Squeeze the potato pulp thus obtained through a muslin cloth and keep it in a flask.
d. Take 20 g of agar in a beaker and warm it with 500 ml of water.
e. Mix both the solution of potato and agar and add 20 g dextrose to it.
f. Thus one litre of PDA medium is prepared.
g. Autoclave the medium at 15 pounds pressure for 15 minutes.

B. Effects of antibiotics on soil microorganisms

1. Take 2 g of soil and dissolve it in 10 ml of water in a beaker. Let the soil particle settle down.
2. Take 5 oven sterilized petridishes and pour 1 ml of soil suspension in each of the plates.
3. Now pour 1ml of the four antibiotics separately in four petridishes with the help of syringe, and
mark them with marker pen.
4. Leave the fifth petridish without antibiotic to serve as control.
5. Pour PDA in each of the petridishes and mix the suspension by rotating the petridishes.
6. Leave the petridishes undisturbed at a warm place for an Observation Colonies of micro-
organisms appear on the culture medium, count the colonies in each petridishes and present
your observation in the following table:

S.No. Antibiotics No. of colonies in a petridish

1. Penicillin NIL
2. Streptomycin 2-3
3. Terramycin NIL
4. Chloromycetin 1-2
5. Control 30-40

The effect of different antibiotics on the micro – organisms can be assessed by counting the number and
size of the colonies growing in the petridishes.
Conclusion
Penicillin and Terramycin was most effective antibiotic against microorganism in soil.
Bibliography
1. Comprehensive practical biology
2. Wikipedia