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ISOLATION AND BIOCHEMICAL CHARACTERIZATION OF
STAPHYLOCOCCUS AUREUS FROM SWEET SAMPLES OF DIFFERENT
AREAS
“Dissertation Report”
Submitted to
GOVT. KAMLA RAJA GIRLS POST
GRADUATE AUTONOMOUS
COLLEGE (JIWAJI UNIVERSITY) GWALIOR
(M.P.)
PRESENTED BY :
CHANDRESH SENGAR
M.Sc IV sem
UNDER THE SUPERVISION OF
Asst. Prof. Ms. PRIYANKA KAMRA
BIMR COLLEGE OF PROFSSIONAL STUDIES,SURYA MANDIR ROAD,
GWALIOR

2. Aim Of The Project
Isolation, Detection, Biochemical
Characterization and Sensitivity Test on
Staphylococcus aureus existing in variety of
sweet samples in differents area.

3. CONTENTS
Introduction
Classification
Morphology and Characteristics
Growth Requirements
Methods and Result
Conclusion
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4. STAPHYLOCOCCUS AUREUS
Staphylococcus aureus was first
observed by Von
Recklinghausen but not isolated.
Sir Alexander Ogston, a Scottish
surgeon, first showed in 1880
that a number of human
pyogenic diseases were
associated with a cluster-forming
micro-organism. He
introduced the name
staphylococcus aureus.
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5. The word staphylococcus is a Greek language “staphyle
means a bunch of grapes” and “kokkos means a round or
granule”.
Staphylococcus aureus is also known as golden staph.
It is a common cause of foodborne illness, commonly called
“Staph aureus”, this is produce a poison or toxin that
cause the illness.
Staphylococcus aureus is a common bacterium found on
the skin and in the noses of up to 25% of healthy people
and animals.
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6. It is important because it has ability to make seven
different toxins that are frequently responsible for food
posioning.
Staphylococcal food poisoning is a gastrointestinal illness.
It is caused by eating foods contaminated with toxins
produced by Staphylococcus aureus.
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7. CLASSIFICATION
Family - Staphylococcacea
Member - Firmicutes
Genus - Staphylococcus
Some species - S. saprophyticus
S. epidermidis
S.hemolyticus
S.ludgunesis
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more
than 36
species

8. MORPHOLOGY AND CHARACTERISTICS
It is gram positive cocci arranged in grape like clusters
forms.
It is a non-motile, non-sporing or catalase positive
bacterium.
It is a single cells in pairs or short chains of three or four
chains.
Its individuals coccus size is 0.5 to 15 μm in diameter.
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9. GROWTH REQUIREMENTS
Its bacteria are able to grow in a wide range of optimum
temperatures is 30 to 37°C.
Its Ph – 7.0 to 7.5.
Its grow on agar based culture medium (like NAM or BP)
and incubated for 24 hours then colonies appears.
The colonies appear to be 2-4mm in diameter.
The colonies appear is circular, smooth or shiny.
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10. SAMPLE COLLECTION
Gole ka mandir – Rasmalai
Hajira – Barfi
Murar – Rabaddi
Thatipur – khoe ka laddu
Kilagate – Rasmalai
Morena – rabaddi
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11. METHODS AND RESULT
1. Isolation Of Bacteria
It is isolated on Nutrient Agar Plate or Baired Parker
Medium.
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NAM plates show
whitish colonies are
BP plates shows
cremish colonies

12. Bacteria grow on specific media is Mannitol Salt Agar
Plates.
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MSA plates show yellowish
colonies are grow.

13. 2. Morphological Identification
• Gram’s Staining – Its smear showing blue colour coccus
are present in bunches like grapes means staphylococcus
are conform and show the colony in 40x.
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Shows gram staining are
positive.

14. 3. Biochemical Characterization
Absence of red colour
was given by bacterial
culture. So
Staphylococcus aureus
are negative.
Red colour was shown by
bacterial culture.
So Staphylococcus aureus
are positive.
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15. VP TEST
No change in colour was
shown by bacterial
culture. So
Staphylococcus aureus are
negative.
Pinkish colour was shown
by bacterial culture. So
Staphylococcus aureus are
positive.
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16. No change in colour of the medium was shown by
bacterial culture. So Staphylococcus aureus are positive.
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17. Gelatinase produce was shown by bacterial culture.
So Staphylococcus aureus are positive.
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Gelatin Hydrolysis test
before adding Mercuric
Gelatin Hydrolysis
test after adding

18. zone of bacterial colonies are diffuse after adding Gram’s
Iodine. So Staphylococcus aureus was negative.
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Starch hydrolysis
test before adding
Starch hydrolysis
test after adding

19. The presence of clear area surrounding the bacterial
growth was shown. So Staphylococcus aureus was
positive.
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After 24 hours of
bacterial colonies
After 48 hours of
bacterial colonies

20. Bubbles were produced by
bacterial culture. So
Staphylococcus aureus are
positive.
No colour change was
given by bacterial culture.
So Staphylococcus aureus
are negative.
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21. Measure of zone of inhibition of antibiotics on NAM media plate.
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Measure of zone of inhibition of antibiotics on NAM media plate.
Resistant antibiotics Resistant zone
Amphicillin 28 mm
Cefazolin 20 mm
Cefotaxime 22 mm
Oxacillin 25 mm
Penicillin 27 mm

22. Result for sensitive
antibiotics-
• Moxiflox
• Sepraflox
• Azymicin
• Lemizal penicillin
• Norfloxa
• Tobra
• Gentamycine
Result for resistance
antibiotics-
• Amphicilline
• Cefazolin
• Cefotaxime
• Oxacillin
• Penicillin
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23. CONCLUSION
We have isolated food posioning Staphylococcus aureus
from different sweets samples of differents areas.
This study indicates that the prevalence of S. aureus
among food handlers was high.
Contamination of food was also high indicating lack of
hygienic practices among the food handlers.
All the isolates were resistance to oxacillin and penicillin.
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