1.

EXPERIMENT 1 : Bacteria ( Hetertrophic eubacteria ) : Are bacteria present

in the lab ?

2.
1.
2.
3.
4.

OBJECTIVE
.Define coccus , bacillus , spirillium , Gram stain
.Describe characteristics of Eubacteria
.Identify and classify the organism studied in this exercise
.Distinguish between Gram positive and Gram negatibe bacteria ,
indicating their susceptibility to certain antibiotics.

3.

INTRODUCTION
The Eubacteria (kingdom Eubacteria) protistan (kingdom Protista) are
among the simplest of living organisms. Both kingdom have unicellular
organism within them , but thats where the similarity ends .
The member of the kingdom Eubacteria are prokaryotic organism ,
meaning that their DNA is free in the cytoplasm ,unbounded by a
membrane . They lack organelles (cytoplasmic structure bounded
by membrane) . By contrast , the kingdom protista consist eukaryotic organism :
The genetic material contained within the nucleus and many
of their cellular components are compartmentalized into membrane
bound organelles.Its likely you examined the distinction between prokaryotic and
eukaryotic cellular organization .
Both eubacteria and unicellular protistan are at the base of the food
chain.From the ecological standpoint , these simple organism are among
the most important organism on our planet.Ecologically , they are much
more important than we are .
KINGDOM EUBACTERIA
Eubacteria are such organism as bacteria and cynobacteria . Most
bacteria are heterptrophic , depend upon an outside source for nutrition
, while the cyanobacteria are autotrophic ( photosynthetic ) , able to
produce their own carbohydrate .

some autotrophic bacteria are pathogens , causing plant and animal

disease , but most are decomposers , breaking down and recycling the waste
products of life . Others are nitrogen-fixers , capturing the gaseous nitrogen
in the atmosphere and making it available to plants via symbiotic association
with their roots.

4.

MATERIALS AND APPARATUS

4.1. dH2O in dropping bottle
4.2 .Four (4) nutrient agar culture plate
4.3. Sterile cotton swab
4.4 .China marker
4.5.Two bottle labeled A and B (A:contain tap water,B:contain 70% ethyl alcohol )
4.6.Tranparent adhesive tape
4.7.Paper towel

5. PROCEDURE
5.1.Four petri dishes containing nutrient agar was obtained . Then by using

china marker , one of the petri dish was label with Dish 1 : Control .
The other was label with Dish 2 : Dry swab , Dish 3 : Treatment A ,
And Dish 4 : Treatment B , also include the group number.
5.2.The sterile cotton swab was run within the lab ( the floor surface ) .
5.3.Then , lift the lid on the Dish 2 as little as is necessary to run the swab
over the surface of the agar .
5.4.The lid was tape securely to the bottom half of the dish by using tranparent
adhesive tape .
5.5.after that , one paper towel was soaked with liquid A and the second paper
towel was soaked with liquid B
5.6.Then , the paper towel was wipped done one-half of the surface that just
sampled ( the floor ) with liqwuid A ( tap water ) , the other half with liquid B
( 70% ethyl alcohol ) . After the area have dried , the dishes 3 and 4 are used
and the procedure in step 2 repeated.
5.7.The culture was placed in a incubator oven to incubate until the next lab
period ( 2 weeks ).
5.8.The bacteria colonies , colour , shape and texture of the bacterial growth
Was examined later .

6. RESULT
FIGURE 1
Bacteria colony on Different Condition

Figure 1.1 : Dish 1 -

Figure 1.3 : Dish 3 Treatment A

(Tap water)

Figure 1.2 : Dish 2 Dry

swab

Figure 1.4 : Dish 4 Treatment B ( 70%

Ethyl alcohol)

7. DISCUSSION
Based on the experiment , there are four type of petri dish was labeled
with control , dry swab , treatment A and treatment B which is to determine the
colonies of the bacteria , the colour and the texture of the growth .

For the control agar plate it was not exposing to the environmental of the
laboratory and seal with the transparent adhesive tape . Based on our result the
the control perti dish was not contaminated .
On the top of that , in the dry swab plate , there are two colonies of the
bacteria was obtained . Both colonies of the bacteria was white in colour and
it has round shape but the other one of the colonies was small in size while the
other is a litle bit bigger . The number of the bacteria prensence for both colonies
only single bacteria.
For the treatment A , which is the tap water was wiped on the surface
of the floor , and then the sterile cotton swab was run on the floor then strike to
the agar plate . From the experiment , the bacteria were highly growth in the
plate .There are 3 colonies of the bacteria . For the two colonies it has white in
colour with round and irregular shaped while the other colonies it has yellow in
colour and randomly shaped . Contamination also occur in this plate.
For treatment B , the surface was wiped with 70% of ethyl alcohol and
and then the strile cotton swab was run on the surface then strike on the agar
carefully . There are two colonies of the bacteria are growth in the plate . Both
are in white colour and has a round shape .Based on the theory , bacteria cannot
growth in high concentration of 70% of ethyl alcohol . However result that we
obtained from the experiment against the theory , we assume that the agar plate
for treatment B already contamninated .
In conclusion , all the agar plate was contaminated except for the the control
plate because its not exposing to the environmental .So , there are several
technique to prevent contamination to occur in agar plate . For example , do not

talk with each other while striking the plate . Beside that , always using aseptic
technique where all the material must be sterilize and petri dish must covered as
much as possible .

8. CONCLUSION

From the experiment , we can conclude that bacteria can growth in dry
swab and treatment A condition .While , bacteria cannot grow in high
concentration of 70% ethyl alcohol. Our objective is achieved where the
morphology of bacteria are indentified have colour , shape and smooth surface .

9. REFFERENCE

9.1 . Campbell , N.A.Reece ,J.B (2008) Biology:9th Edition

9.2 . Solomon , E.P Berg L.R & Martin , D&W (2008) Biology: 9th
Edition.Thomson
9.3. Manual Amali , BIO320

1. EXPERIMENT 2 : Bacteria ( Hetertrophic Eubacteria ) : Bacteria shape

And sensitivity of antibiotics

2. INTRODUCTION
Simple stain can be used to determined the cell shape , size and
arrangement .
Bases on its name simple stain is very simple staining procedure involving
only one
Stain . It can be methylene blue , gram safranin or gram crystal violet .
This stain will readily give up a hydroxide ion and accept a hydrogen
ion in which leave the stain positively charged. Since the surface of most
bacteria cell is negatively charged these positively charged stain

will adhere readily to cell surface .

Gram staining is staning technique that used to distinguish
between gram positive and gram negative bacteria which have
consistent different cell wall.Gram positive bacteria stain purple colour
and Gram negative bacteria will stain pink-red colour

3. OBJECTIVE
3.1 .Distiguish between the gram-positive and gram-negative bacteria
properly .
3.2 .To develop the laboratory skill in gram staining in proper way.
3.3. To observe the shape of the bacteria ( coccus ,bacillus and spirillum ).

4. MATERIAL AND APPRATUS

PREAPRATION OF SMEAR , SIMPLE STANING AND GRAM STAINING
5.1.Bacterial culture
5.2.Inoculation loops
5.3.Bunsen burner
5.4.Glass slide
5.5.Methylene blue
5.6 .Gram stain ( crystal violet , iodine , safranin , alcohol )
5.7.Compund light microscope
5.8.Immersion oil
5.9.Handphone camera
5.10.Tissue paper
5.11.Distilled water
5.12.Tap water

5. PROCEDURE
5.1.SMEAR PREPARATION
Using aseptic technique a loopful of the bacterial culture was tranfered
onto a clean glass slide and spread the culture evently to form
a good thin smear.
The smear allowed to be air-dry.
Once the smear properly dried , it was fix-heated.By holding one
end of the slide and the slide run across the flame for 2-3 times.
The heat-fixed smear was later allowed to be cool to room temperature
for a few minute .
5.2.SIMPLE STAINING ( METHLYNE BLUE )
The smear was flood with methylene blue for 1 minute .
Excess stain was removed from the smear.
Grntly , the smear was rinsed under slow running tap water.
The stained was allowed to air-dry before observing under microscope
The observation was recorded.

5.3.GRAM STAINING
Heat-fixed smear was flood with crystal violet for 1 minute . The excess
crystal violet was removed and rinsed with iodine
The smear flood with iodine for 1 minute and excess iodine was removed.
The smear later was decolourise by running acetone alcohol down the smear
for a few second .
Immediately the smear was rinse under slow running tap water until all trace
alcohol was removed

 After that , the smear was flood with safranin for only 1 minute then excess
safranin was removed and rinsed the smear for the last time under running tap
water .
The smear was blot dry by using paper towel
The preparation was examine under microscope and the observation was
recorded.

6. RESULT

7. DISCUSSION

8.CONCLUSION
From the experiment that we have been conducted , we can say
that the we need to stain the bacteria in oder to see their structure
clearly under microscope . We can use simple staining and also gram
staining method for to be more specific . In this experiment we were
enable to determine the type of bacteria based on their shape ,
arrangement and colour .

9.REFFERENCE

9.1 . Campbell , N.A.Reece ,J.B (2008) Biology:9th Edition

9.2 . Solomon , E.P Berg L.R & Martin , D&W (2008) Biology: 9th
Edition.Thomson
9.3. Manual Amali , BIO320
9.4. Prescott.L(2002).Procaryotic cell structure and function . In Microbiology
(5th.ed..pp 60-61).New York.NY 10020.The MacGraw-Hill Companies.

UNIVERSITI TEKNOLOGI MARA

BIO 320
AS1204A2
PRACTICAL 1 : EUBACTERIA
GROUP 3
NAME : NURSYAHIRAH BINTI SUHAILI
ID NUMBERS : 2015214946
LETCURER NAME : SIR ANSIR SALEH