B2231 – Microbiology Lab S1 2020/2021

Group:

Name: Christopher Baaklini

ID: 57396

1. Introduction of microbiology laboratory

1.1 Safety instructions
Wearing a buttoned lab coat
Avoid eating and bringing fingers near mouth
Manipulations of bacteria within range of a Bunsen burner flame (10-15 cm)
Avoid placing flammable objects near flame
No pipetting to the mouth
Heat metal loops before and after manipulation to desiccate remains of culture
Sterilize all equipment after experiment

1.2 Different conditions of sterilization to work in aseptic conditions

Disinfection of benchtop using Dettol
Heat sterilization using a Bunsen burner

1.3 Identification of saprophytic and commensal microorganisms

1.3.1 Aim of these two experiments
To distinguish between both types of bacteria and observe their different characteristics

1.3.2 Materials and methods

Petri dishes containing agar, pipette, slides and cover slips, microscope.

1.3.3 Results obtained after observation on the petri dishes

a) Saprophytes microorganisms
Hand bacteria:
Size: 0.4 to 0.5 mm
Form: most are circular, some are irregular
Aspect: smooth
Opacity: opaque
Consistency: creamy
Color: pale yellow
Ear bacteria:
Size: 0.1 to 0.2 mm
Form: curved
Aspect: smooth
Opacity: opaque
Consistency: creamy
Color: white
 b) Commensal microorganisms
Size: 0.3 to 0.5 mm
Form: flat
Aspect: smooth
Opacity: translucent
Consistency: dry
Color: yellow-white

1.3.4 Conclusion
Hand and ear bacteria are saprophytic while that acquired from the environment are
commensal.

1.4 Description of medium used

Mannitol salt agar

Differentiates pathogenic Staphylococcus from Micrococcus since the level of NaCl is inhibitory
to most other bacteria.

Nutrient agar
Contains mammalian blood at a concentration of 5-10%.

MacConkey
Selective media that isolates Gram-negative bacteria and inhibits Gram-positive organisms by
the bile salts and crystal violet. Lactose fermenting colonies appear red or pink while non-
lactose fermenters are colorless.

2. Gram staining
2.1 Objective of this experiment
Distinguish and classify bacteria according to properties of their cell wall.

2.2 Principle of this experiment

Gentian violet binds to cytoplasmic components. After staining, all bacteria appear purple. In
gram-negative bacteria, the peptidoglycan rich wall allows the passage of acetone which
discolors the cytoplasm whereas in gram-positive bacteria, the wall constitutes of an
impermeable barrier and the cytoplasm remains purple.

2.3 Material and method

Seeded petri dishes, inoculation loop, slides, sterile distilled water, Gram stains.

2.4 Result
 Violet bacteria are obtained on the slide with a spherical shape.

2.5 Conclusion
The bacteria are Gram-negative with a coccus shape.