MANGALPUR, BALURGHAT, SOUTH
DINAJPUR, WEST BENGAL
  PROJECT REPORT SUBMITTED BY
          SUBHORAJ SAHA
             CLASS – XII
             ROLL NO. –
YEAR – 2019 – 2020 {CLASS XII SESSION}
SUBJECT –BIOLOGY [SUB CODE –044]
 Bona fide certificate
This is to certify that this project entitled“study of
drug resistance in bacteria using
antibiotics”by Subhoraj Saha of class-xii,
Roll no. – _____________ ,
Registration no. – ____________ ,
Academic session – 2019-2020
Submitted in fulfillment to ‘ class – xii biology (044)
project’ , is bona fide record of work carried out
under my guidance and supervision.
                                        ______________
                                            Signature
              Name – suparna Bhattacharjee
                                      Designation – PGT
                                    Department – Biology
         School – Techno India Group Public school
                             Date        – ______________
Acknowledgement
1) I would like to express my special thanks
of gratitude to my biology teacher
 “Mrs. –suparna Bhattacharjee” for
their able guidance and support in completing
my project.
2) I would also like to extend my gratitude to
my parents for providing support and
helping me in completing the project.
                             ________________
                          Signature of candidate
                           Subject –Biology
                          Subject code – 044
                           Roll no. – ___________
 Contents
SR.NO      TITLE                 PAGE
                                  NO
  1)        Introduction          1-5
  2)    History of antibiotics     6
  3)       What is drug            7
            resistance?
  4)     Why do bacteria           8
        become resistant to
           antibiotics?
  5)      Aim of project           9
  6)          Materials          10 - 12
  7)         Procedure             13
  8)          Method             14 - 15
  9)        Observation            16
 10)           Result              17
  11)        conclusion            18
 12)         precaution            19
 13)        bibliography           20
Introduction -
 Antibiotics are drugs that are used in the
    treatment or prevention of bacterial
infection. Strictly speaking ,antibiotics are
       natural substance produced by
microorganisms to kill other organisms or
            retard their growth.
Penicillin, Tetracycline, Streptomycin are
few drugs used to treat various diseases.
HISTORY OF ANTIBIOTICS
                ANCIENT
In 3500 BC,the Sumerian doctors would give
their patients,beer soup with snakeskin and
turtle shells.
Modern History -
 Louis Pastures was the first to recognize that
 bacteria could be used to kill other bacteria.In
     1929 Sir Alexander Fleming discovered
 Penicillum Notatum.It is effective again range
   of bacteria.Then Streptomycin discovered.
  What is drug
   resistance ?
Antibiotics resistance occurs when an
antibiotics has lost its ability to effectively
control or kill bacterial growth; in other
words, the bacteria are”Resistance”and
continue to multiply in presence of
therapeutic levels of an antibiotic.
          Why do bacteria
    become resistant to
                antibiotics?
Antibiotics resistance is a naturalphenomenon. When an
antibiotics is used, bacteria that can resist that antibiotics
have a greater chance of survival than those that are
“Susceptible”.
Susceptible bacteria are killed or inhibited by an antibiotics,
resulting in a selective pressure for the survival of resistant
strains of bacteria.
In some countries and over Internet, antibiotics can be
purchased without a doctor’s prescription. Patients
sometimes take antibiotics unnecessarily, to treat viral illness
like common cold.
Aim of the
  project-
In this project the aim is to
  investigate the Study of
    development of drug
resistance in bacteria using
         antibiotics.
Materials-
BUNSEN BURNER   FORCEPS
  SOAP                    PARAFILM
              ISOPROPANOLCOTTON SWAB
MATCHES   BROTHCULTURES
                          OF
                     S.EPIDERMIDIS
TSA PLATES            BENT GLASS ROD
FINE TIPPED MARKING     ANTIBIOTICS
IMPREGNATD                   DISCS
Procedure-
It involes the following steps :
To compare how effective one effective one
antibiotic is to another, or to measure the degree
of antibiotic resistance in a bacterium a procedure
called the Kirby-Bauer test can be done.
To do this,
  1) A pure strain of bacteria is isolated from an
       infected person.
  2) This pure strain is then spread over the surface of a
       special medium ,called Mueller-Histon Agar ,to
       create a lawn ,or carpet of bacteria.
  3) Small filter paper discs ,impregnated with
       standardised amounts of antibiotics ,are gently
       pressed on to the surface of the agar.
  4) The plates are then incubated overnight ,the
       antibiotics diffuses from the disc and into the
       agar.This antibiotics diffusing into the agar will
       inhibit the growth of susceptible bacteria.
Method -
   1) Clean your work area and sterilize it with
                    70% isopropanol.
    2) Swirl the contents of the brothculture of
          S.Epidermis until it is equally murky
      throughtout.Use the sterile cotton swab and
         glass rod to create a lawn ,or carpet of
         S.epidermis on 2 of the plates of TSA.
Don’t forget to hold the cap in your pinky and to
flame the mouth of the test tube before and after
         dipping the sterile swab into it.
3)Label the bottom of petric dish with your name
 ,the date be sure to write small and only around
        the edges of the bottom petri dish.
4) Use the second sterile swab to repeat step two
  ,expect substitute E.coli for S.epidermis. Also
              ,label this E.Coil dish.
  5) Select two different antibiotics discs. Place
 them on oppsite side of a petri dish containing
 S.epidermis ,with the code side facing up. T ap
them gently with sterile forceps to stick them to
                     the agar.
 6) Using the second plate ,repeat step 5 ,again
using two different antibiotics discs. You should
have four different antibiotics discs on two plates
                 of S.epidermidis.
7) Repeat steps 5 and 6 using the plates of E.coli.
Be sure to use the same types of antibiotics discs
          as were used in steps 5 and 6.
8) Warp parafilm around all four plates and place
  the plates on their lids and in the incubator at
                 37˚C for 24 hours.
9) After 24 hours in the incubator ,check for the
         presence of antibiotics activity.
Observations -
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 CLEAR
AREAS.THESE ARE THE COLONIES OF RESISTANT
STRAINS OF THE BACTERIA.
                    .
CONCLUSION:-
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.
 PRECAUTIONS-
A microbiology lab is potentially a very dangerous
place.For this reason it is extermely important
that you follow safety guidelines and always
practice sterile technique when handling microbes
,unless instructed otherwise.
   1) There should be no books or paper at your
        workstation except this lab packet.
     2)Never have any food or drinks at your
                 workstation.
     3)Never open a petri dish after you have
  inoculated it and have allowed it to incubated
                    overnight.
4) Always dispose of used material in the biohazer
        bag unless instructed otherwise.
BIBIOLGRAPHY –
       https://en.wikipedia.org/wiki/Drug_resistance
        https://aidsinfo.nih.gov/understanding-hiv-
             aids/glossary/212/drug-resistance
https://www.niaid.nih.gov/research/antimicrobial-resistance-
                        definitions
 https://www.sciencedirect.com/topics/neuroscience/drug-
                       resistance
https://www.medicinenet.com/script/main/art.asp?articlekey
                         =3120