ENVE 202 Environmental Microbiology Lab Report

Chemical Agents of Control: Chemotherapeutic Agents (Kirby-Bauer Method & Synergistic Effect of
Drug Combinations)
Name: İdilay KONAR
Date:11.4.2020 Section: 2.2

Observation and Results:

1. Record the zone size and susceptibility of each test organism to chemotherapeutic agent as
resistant (R), intermediate (I) or sensitive (S) in the charts below. (Check the Table 2 on page 309
for the classification. Make sure to use the appropriate values for Penicillin and Vancomycin
according to the microorganisms. )

GRAM-NEGATIVE
E.coli P. plecoglossicida
Chemotherapeutic Zone size Susceptibilit Zone size Susceptibility

m
er as
Agent (mm) y (mm)

co
Penicillin 6 R 0 R

eH w
Streptomycin 28 S 28 S

o.
Tetracycline 22 S 16 I
Chloramphenicol rs e 37 S 27 S
ou urc
Gentamicin 32 S 25 S
Vancomycin 0 R 0 R
o
aC s
vi y re

GRAM-POSITIVE
M. luteus E. faecalis
Chemotherapeutic Zone size Susceptibility Zone size Susceptibility
Agent (mm) (mm)
ed d

Penicillin 16 R 0 R
ar stu

Streptomycin 24 S 26 S
Tetracycline 10 R 15 I
is

Chloramphenicol 26 S 26 S
Gentamicin 12 R 25 S
Th

Vancomycin 0 R 16 I
sh

2. For each of the chemotherapeutic agents, indicate the following

1
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 a. The spectrum of its activity as broad or limited ( If more than 2 m.o. are susceptible to
the antibiotic classify it as broad otherwise limited)
b. The type or types of organisms it is effective against as gram-positive or gram-negative
or both.

Chemotherapeutic Spectrum of activity Type(s) of

Agent microorganisms
Penicillin Limited Non effective

Streptomycin Broad Effective for both

Tetracycline Broad Effective for gram

positive and few gram
negative
Chloramphenicol Broad Effective for both

Gentamicin Broad Effective gram negative

m
and gram positive

er as
Vancomycin Limited Effective for gram

co
positive

eH w
o.
3. For “Appearance of zone inhibition” write down bridge forming (merging) or separate depending on the
rs e
diameters. In the actual experiment you could have just observed whether the zones are merging or not,
ou urc
but the given diameters should give you an idea if they are merging. Check the disc placement distances on
the manual and compare it to the values provided in this table.
!!! Remember that these discs are placed at a certain distance (check example for E.coli below) from the
o

centers and given values are for diameters.

aC s
vi y re

Trimethoprim Sulfisoxale
12.5 mm 12.5 mm
ed d
ar stu

Diameter of
Cultures & Agents Zone Inhibition Appearance of Synergistic or Additive
is

(mm) Zone inhibition Effect

Th

E.coli:

Trimethoprim and sulfisoxazole 28 – 36 Merged Synergistic effect

Trimethoprim and tetracycline 18 – 20 Separated Additive effect

sh

S. aureus

Trimethoprim and sulfisoxazole 32 – 36 Merged Synergistic effect

Trimethoprim and tetracycline 20 - 22 Separated Additive effect

2
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 Review Questions
1. Your experimental results indicate that antibiotics such as tetracycline, streptomycin and
chloramphenicol have a broad spectrum of activity against prokaryotic cells. Why do these
antibiotics lack inhibitory activity against eukaryotic cells such as fungi?

The reason for this may be because these antibiotics inhibit protein synthesis. The reason for lack of
inhibition caused by the difference between ribosomal components 70S for prokaryotes and 80S for
eukaryotes.

ENVE 202 Environmental Microbiology Lab Report

Chemical Agents of Control: Disinfectants and Antiseptics
Name: İdilay KONAR
Date: 11.4.2020 Section:2.2

m
Observation and Results:

er as
1. Indicate the absence of a zone of inhibition as (-) and the presence of a zone of inhibition as (+) at

co
eH w
Activity column. (If the zone of inhibition is larger than uninoculated control then it could be
accepted as present).

o.
Microbial Zone of
rs e Antimicrobial Agent
ou urc
Species Inhibition for Tincture of Iodine 3% Hydogen 70% Isopropyl 5% Chlorine Bleach
control (mm) Peroxide Alcohol
Zone of Activity Zone of Activity Zone of Activity Zone of Activity
Inh(mm Inh(mm Inh(mm) Inh(mm)
o

) )
aC s

E.coli 7 9 + 23 + 15 + 13 +
vi y re

S.aureus 8 8 - 15 + 11 + 11 +
ed d

2. Which of the experimental chemical compounds appears to have the broadest range of microbicidal
ar stu

activity and the narrowest range of microbicidal activity?

The broadest range of microbicidal activity: 3% Hydogen Peroxide.

Narrowest range of microbicidal activity: Tincture of Iodine.
is
Th

Review Questions

1. Can the disinfection period (exposure time) arbitrarily increase? Explain.

sh

Exposure time depends on the bacteria. Exposure period can be increased or decreased if
necessary. Environmental conditions must be considered while altering exposure time.
2. A household cleanser is labeled germicidal. Explain what this means to you.

Germicides are able to destroy germs.

3
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