STUDY OF DRUG

RESISTANCE USING
ANTIBIOTICS
CERTIFICATE OF AUTHENTICITE
This is to certify that “ swati Chhachhia” a
student of class XII B has successfully
completed investigatory project under the
guidance of Mrs. Manju Phatak .

Signature External
(subject teacher) Examiner
 ACKNOWLEDGEMENT
I feel proud to present my investigatory project in Biology
on the “STUDY OF DRUG RESISTANCE IN BACTERIA USING
ANTIBIOTICS” . This project would not have been feasible
without the proper rigorous guidance of biology teacher
Mrs. Manju Phatak who guided me throughout this project
in every possible way . An investigatory project involves
various difficult lab experiments, which have to obtain the
observations and conclude the reports on a meaningful
note. Thereby, I would like to thanks Mrs. Manju Phatak for
guiding me on a sustematic basis and ensuring that in
completed all my research with ease. Rigorous hard work
has to put in this project to ensure that it proves to be the
best. I hope that it proves to be the best. I hope that this
project will to be proves to be a breeding ground for the
next generation of students and will guide them in ever
possible way .
 INTRODUCTION
An antibacterial is an agent that inhibits bacterial
growth or kills bacteria. The term is often used
synonymously with the term antibiotic(s). Today,
however, with increased knowledge of the causative
agents of various compounds, including anti-fungal and
other compounds. Antibacterials must be distinguished
from disinfectants(sanitizing agents), which are less-
selective substance used to destroy microorganisms.
The term antibiotic was first used in 1942 by Selman
Waksman and his collaborators in the journal articles to
describe any subatance produced by a microorganisms
that is antagonistic to the growth of other
microorganisms in high dilution. This defination
excluded substances that kill bacteria but are not
produced by a microorganisms (such as gastric juices
and hydrogen peroxide). It also excluded synthetic anti
bacterial compounds such as the sulfonamides. Many
antibacterial compounds are relatively small molecules
with a molecular weight of less than 2000 atomic mass
units.
 CLASSES
Antibacterial antibiotics are commonly classified based on their
mechanism of action, chemical structure, or spectrun of activity .
Most targer bacterial functions or growth processes. Those that
target the bacterial cell wall (penicillins and cephalosporins) or
the cell membrane (polymyxins), or interfere with essential
bacterial enzymes (rifamycins, lipiarmycins, quinolones, and
sulfonamides) have bactericidal activities. Those that target
protein synthesis (macrolides, lincosamides and tetracyclines) are
usually bacteriostatic (with the exception of bactericidal
aminoglycosides)
MEDIAL USES
TREATMENT
•Bacterial infection
•Protozoan infection e.g.Metronidazole is
effective against several parasitics
•Immunomodulation e.g.Tetracycline is effective
against periodontal inflammation, &
Dapsone which is effective against autoimmu-
-ne diseases oral mucous membrane.

PREVENTION OF INFECTION
•Surgical wounds
•Dental antibiotic prophylaxis
•Conditions of neutropenia e.g. Cancer-related
 SIDE EFFECTS
Antibacterial are screened for any negative
effects on humans or other mammals before
approval foe clinical use, and are usually
considered safe and most are well-tolerated. However,
some antibacterials have been associated with a
rangeof adverse effects. Side-effects range from mild
to very serious depending on the antibiotics used, the
microbial organisms targeted, and the individual patient.
Safety profiles of newer drugs are often not as well-
established as for those that have a long history of use.
Adverse effects range from fever and nausea to major
allergic reactions, including photodermatitis and
anaphylaxis. Common side-effects include diarrhea,
resulting from disruption of the species composition in
the intestinal flora, resulting, for example, in
overgrowth of pathogenic bacteria,such as Clostridium
difficle. 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 frominteraction with other drugs,
such as elevated risk of tendon damage from
administration of a quinolone antibiotic wit a systemic
corticosteroi
 Antibacterial-resistant strains and species, sometimes
referred to as “superbugs”, now contribute to the
emergence of diseases that were for a while well-
controlled. For example, emergent bacterial strains
causing tuberculosis(TB) that are resistant to
previously effective antibacterial treatments pose many
therapeutic challenges. Every year, nearly half a million
new cases of multidrug-resistant tuberculosis(MDR-TB)
are estimated to occur worldwide. For example, NDM-1
is a newly identified enzyme conveying bacterial
resistance to a broad range of beta- lactam
antibacterials. The United Kingdom's Health Protection
Agency has stated that “most isolates with NDM-1
enzyme are resistant to all standard intravenous
antibiotics for treatment of severe infection”.
 RESISTANT PATHOGEN
ACINETOBACTER
Acinetobacter is a group of bacteria commonly found in
soil and water . While there are many types or “species”
of Acinetobacter and all can cause human disease,
Acinetobacter baumannii accounts for about 80% of
reported infections.
Outbreak of Acinetobacter infections typically occur in
intensive care units and healthcare settings housing
very ill patients. Acinetobacter infection rarely occur
outside of healthcare setting.
 ANTHARX
Antharx is a serious disease caused by Bacillus
anthracis, a bacterium that forms spores. Antharx most
commonly occours in wild and domestic mammalian
species, but it can also occour in humans when they are
exposed to infected animals or to tissue form infected
animals or when anthrax spores are used as a
bioterrorist weapon. Some starins of B. Anthracis may
be naturally resistant to certain antibiotics and not
other. In addition, there may be biologically mutant
strains that are engineered to be resistant to various
antibiotics.

CAMPYLOBACTER
Campylobacter is estimated to cause over 1.3million
infections and 76 death in the United States each year.
Many of these infections are foodborne. Campylobacter
infections can result in long-term consequences, such as
arthritis or a type of paralysis called Guillain-Barre
syndrome.
GROUP B STREPTOCOCCUS

Group B Streptococcus (group B strep) is a type of

bacteria that causes illness in newborn babies, the
elderly, and the adults with other illness , such as
diabetes or liver diseases . Group B strep has shown
confirmed resistance to certain antibiotics
 DO'S AND DON'TS OF
ANTIBIOTICS
•DO take the antibiotic exactly as indicated on the
container.
•DO finish the prescription even if you being to feel
better.
•DON'T demand antibiotics when your health care
provider says they are not needed.
•DON'T skip doses.
•DON'T share the prescription with anyone else.
•DON'T save the prescription for later time.
MATERIAL REQUIRED
•Sterilised petridishes
•Sterilised culture tube with media
•Transfer loops
•Forcep
•Flask
•Beaker
•Burner
•Penicillin
•Aureomycin
•Hay
•Alcohol
•Agar
•Starch
•Distilled water etc.
EXPERIMENTAL PROCEDURE
•To 200ml distilled water in a flask, add 8gms of agar
powder and 2gms of starch. Put a few pieces of dry hay
into the medium. Cover flask with an inverted beaker.
Boil the medium for 5min and then cool the medium to
room temperature. Place the flask in warm place. Within
2-3days, formation of scum of cloudy suspension appear
on the medium indicating the growth of Bacillus subtilis.
•Take culture tubes with agar medium and heat the test
tubes in warm water to melt agar. Cool each test tube
so that you can hold it in your hand and the agar
remains liquid. Remove the cotton plug and pass the
mouth of the test tube through the burner flame twice.
Flame the transfer loop after dipping it in alcohol and
let it cool. Pick up a loop full of bacterial culture from
the flask and transfer it to the warm agar in the
culture tube. Flame the loop and the mouth of the
culture tube and replace the cotton plug. Roll the
culture tube of warm agar between plams to mix the
bacteria well with agar. Transfer of bacteria should be
done as quickly as possible.
•Take sterilised petridishes. Remove the cotton plug
and flame the mouth of the culture tube. Lift the cover
of the petridish at an angle of 45degree and quickly
pour the medium of the culture tube into the bottom
half of the dish. Remove the culture tube and replace
the cover of the petridish. Move the covered petridish
along the table top in a figure eight pattern to
distribute the medium evenly. Allow the agar to cool.
Prepare two petridishes and mark them A and B.
•Prepare the penicillina nd aeromycin solution by
dissolving the powdered drugs in distilled water. Cut
discs of filter paper of 1cm diameter. Soak a disc in
each of the penicillin and aeromycin solution. Dip the
forceps in alcohol and pass the forceps tip quickly over
the burner flame. Use the sterilised forceps to put
penicillin and aeromycin soaked discs at two distant
sites of petridish A. Consider the petridish B as
control. Keep both the petridishes undisturbed in warm
place or in an incubator to allow the bacteria to grow .
Observe the petridish for several days.
OBSERVATION
The area around the antibiotic discs in the petridish will
be clear. In other areas, colonies of bacteria will be
observed. Examine the clear area in each petridish for
few more days. A few very small colonies may appear in
the areas. These are the colonies of resistant strains
of the bacteria.
CONCLUSIONS
Antibiotic drugs kill most of bacetria strains, hence the
areaa appear clear. However, a few strains which were
resistant in the bacterial population survived and
produced colonies later.
This proves the resistant starin to antibiotics were
present in the bacterial population.
RESISTANCE
SEM depiciting methicillin – resistance Staphylococcus
aureus bacteria.
The emergence of resistance of bacteria to
antibacterial drugs is a common phenomenon. Emergence
of resistance often reflects evolutionary processes
that take place during antibacterial drug therapy. The
antibacteria; treatment may select for bacteria; strains
with physiologically or genetically enhanced capacity to
survive high doses of antibacterials. Under certain
conditions, it may result in preferential growth of
susceptible bacteria is inhibited by the drug. For
example, antibacterials selection for strain having
previously acquired antibacterial-resistance genes was
demonstrated in 1943 by the Luria-Delbruck
experiment. Antibacterials such as penicillin and
erythromycin, which used to have high efficacy against
many bacterial species and strains,have become less
effective, because of increased resistance of many
bacterial strains.