Unknown Bacteria

INTRODUCTION:

In this laboratory exercise explore the differences of microorganism and continue our use of specialized media and use some biochemical testing.This report discloses the basic laboratory instruments will be used in each of our practices .It is of great importance to recognize and identify the different instruments and laboratory tools, because in this way will we be able to use them properly and also to call them by name and know why.
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The purpose of this is knowing the types of bacteria that exist in our environment and diseases caused by them, and how treat it.

MATERIAL AND METHODS:

An unknown bacterium was handed out
by the lab instructor. The methods that have been learned so far in identifying bacteria were applied to this unknown. Procedures were followed as stated in the lab manual and biochemical test handouts.

Lab day 1: The first day in lab I obtained the unknown mixed culture and BHIA, is a liquid medium used in the cultivation of microorganisms, the procedure that was the preparation of this medium, followed step used the septic technique to transfer a small amount of culture with a flame-sterilized inoculating loop to the first quadrant TSA , after that repeat this process with MSA plate, is high concentration of salt selects for members of the genus staphylococcus some organisms may grow, but typically grow very weakly, and then repeated this with the other media procedimeinto untill all media are properly streaked (EMB,PR,TISA,).

Finally incubate the selevtive and differential media for 24 hours.

RESULTS:

In the second day lab, I collected the medium and began by identifying the morphology and cell-to cell arrangements of the colonies. One colony observed, the colony was yellow in color and larger in size. As instructed, the colony was prepared for gram staining in falowing steps:

• The bacteria was smeared on a slide and allowed to dry. • The bacteria was heat gixed to the slide • Added to the smear crystal violet pigment for 1 minute • Wash it with distilled water • Apply gram’s iodine for 1 minute • Wash iodine off with distilled water • Decolorize the smear with alcohol drop until it runs off clear • Wash off alcohol with distilled water • Counterstain with safranin for 45 seconds • Wash off sagranin with distilled water and • Blot dry with bibulous paper
The results are also shown in a flow chart form:

TABLE : CHARACTERISTICS OF UNKNOWN BACTERIA

| TEST | OBSERVATIONS | RESULT |
|TSIA |COLOR BEFORE :PINK RODS |YELLOW |
|BHI |COLOR BEFORE :YELLOW |CLOUDY YELLOW NO GROWTH |
|EMB |COLOR BEFORE: PURPLE |NO CHANGED ,NO GROWTH |
| |BLACK | |
|TSA |COLOR BEFORE: WHITE |GROWTH |
|MSA |COLOR BEFORE:RED LIGHT |GROWTH/NON-FERMENTATION |
|PHENOL/ SUCROSE |COLOR BEFORE: RED |YELLOW |
|GRAM STAIN |COLOR BEFORE:PINK |POSITIVE/COCCI
|

TABLE 2
[pic]
FLOWCHART

Staphylococcus epidermidis

[pic] TSA deeps Faculative anaerobic
Aerobic

+/+ Mannitol/High Salt _/+

[pic]
-----------------------
Staphylococcus epidermidis

Nutrient Agar
Growth

Catalses
+

Oxidase _-

+ Catalase test --

Gram positive cocci.
1)Micrococcus luteus
2)Staphylococcus aureus
3)Streptococcus faecalis
4)Streptococcus lactic

Streptococcus faecalis
Streptococcus lactic

Micrococcus luteus
Staphylococcus aureus
Staphylococcus epidermidis

Micrococcus luteus

Staphylococcus aureus
Staphylococcus epidermidis

Staphylococcus aureus

Staphylococcus epidermidis