Title: Microbiology

Obejctive:
1.To obtain isolated colonies of a pure or mixed culture.
2. To compare the effect of handwashing on bacteria on thumb.
3.To determine gram (+) or gram (-) bacteria.

Procedur:
A)Streak plate technique
1.The inoculating loop was sterilize in the Bunsen burner by putting the loop
into the flame until it is red hot. Allowed it to cool.
2.An isolated colony was picked from the agar plate culture of E.coli and
spread them over the first quadrant on separate agar plate.
3.The agar plate was covered with the lid and the loop was flamed.
4.The plate was turned and lightly streaked into the next quadrant without
overlapping the previous streak.
5.Step 3 and 4 was repeated into the third quadrant.
6.Plate was sealed with parafilm.
The plate was inverted and incubated at 37C for 24 hours.

B) Effect of handwashing on bacteria on thumb

1.4 nutrient agar plate was obtained and labelled :
a)Control
b)Water
c)Hand sanitizer
d)Soap
2.Each agar plate was divided into 4 sections by drawing line using marker
pen on the back of the petri dish
3.Using aseptic technique, gently press thumb on the control agar plate.
4.Wash hands(including thumb) with water and step 3 was repeated on the
appropriate agar.
5.Step 4 was repeated using hand sanitizer and soap.
6.Plate was sealed with parafilm.
7.Plate was inverted and incubated for 24 hours.
Part 2 : Gram staining
1.Using a sterile inoculating loop, 1 drop of sterile water was added to the
slide. Prepared smear of :
a)Escherichia coli
b)Staphylococcus aureus
2.Air dry and heat fix.
3.The smear was covered with crystal violet (primary stain) for 1 min.
4.The slide was gently washed with water.
5.Grams iodine (mordant) was added for 1 min.
6.Wash with water.
7.Decolorize with 95% ethanol.
8.The smear was covered with Safranin for 30 sec.
9.Both the top and bottom of the slide was washed with water.
10.The slide was blotted.
11.Using the light microscope, view the smear up to 100x with immersion oil.
Result

Streak plate technique

Effect of handwashing
Gram Staining
 Pink-gram staining negative

Purple- gram staining positive

Discussion
Part a:streaking
During the experiment for part a(streaking) loop is used to obtained the e.coli.
when it is done from an agar plate with isolated colonies and is transferred to
a new agar plate using a sterile Then,the loop is streaked on the agar surface
like the photo above.while streaking on the agar it should be slow and gently
so the agar will not damage on the agar.last thing is the sample of streaked
need to place in incubated at 37C for 24 hours.

Effect of handwashing
Hand sanitizer is the best to protect our hand(least bacteria) from bacteria
compared to the others(in the picture above)

Part b:Gram staining.

We will asked to prepare two own sample,during preparing the sample the
apparatus and chemical material need to handle very carefully and follow the
procedure given to avoid anything the bad things happen.other than that,the
immersion oil is used to very briefly recap,NA relates to the light gathering
properties of the optical components of microscope, whereas resolution ability
to distinguish details within specimen. Using an immersion lens and oil can
improve both the resolution and NA.another thing is Use more accurate
apparatus so as to decrease percentage errors

Conclusion
The conclusion is the streak plate method is a rapid qualitative isolation
method. This technique is commonly used for isolation of discrete colonies
initially require that the number of organisms in the inoculums be reduced.
Other than that, the results from the handwashing experiment show that
washing with sanitizer is effective compared with others such as water and
soap. Moreover, we have identified that negative-gram and positive-gram are
Escherichia coli and Staphylococcus aures respectively from the gram
staining method.

Reference
1.https://bitesizebio.com/23421/the-why-and-how-of-oil-immersion-
microscopy/
2.https://www.google.com/search?
q=Numerical+Aperture&oq=Numerical+Aperture&aqs=chrome..69i57j0l5.783j
0j7&sourceid=chrome&ie=UTF-8