Lab 5 – Isolation Techniques

Ninoska Garcia-Ortiz 063 053 2

Objective:

• To understand the principle of bacterial isolation

• To recognize the difference between the streak plate and the pour plate
• To understand how to use the two quantitatively
• To be able to write a description of some bacterial colonies
• To be able to identify types in a mixture

TABLE 1 - Isolation by Streaking Results/Discussion

BACTERIA
Staphylococcus epidermis
Escherichia coli
Bacillus subtilis
Pseudomonas seruginosa
Serratia marcescens
Mix 1
MORPHOLOGY
Round scappled, light beige, flat raised spreading edge.
Lobate, light beige, irregular, irregular and spreading.
Irregular spreading, irregular lobate
Transparent colonies
Round scappled, red, flat
Escherichia Coli (E.C)

Bacillus subtilis (B.S)

Mix 2 Serratia marcescens (S.M)

TABLE 2 - Isolation by Pouring Results/Discussion

PLATE DESCRIPTION/OBSERVATION
3 'sprinkles', blotches, round colonies, many growing into the agar
Mix1 diluted X 103
4 Growing into agar, irregular, round, sprinkles
Mix1 diluted X 104
5 Growing into agar, irregular, round, sprinkles
Mix1 diluted X 105
6 Relatively little growth, growing into agar , sprinkles
Mix1 diluted X 106
7 Irregular, spreading, moderate growth, growing into agar, sprinkles
Mix1 diluted X 107

Mix 1 : (Staphylococcus epidermis (S.E), Escherichia Coli (E.C), Bacillus subtilis (B.S)
Discussion:

Isolation by streaking:

- Table 1
- Formation of colonies was successfully achieved in all seven plates.
- In mix 1, only 2 of the 3 colonies were recognized based on their morphology:
Escherichia Coli (E.C) & Bacillus subtilis (B.S).
- In mix 2, only one of the three colonies was recognized based on their morphology:
Serratia marcescens (S.M)
- One possible explanation could be due to the original number of microorganism in the
sample.
- The fact that some bacteria are more fastidious than others could be another explanation.
- Eg. E. coli is non-fastidious so it can grow more easily than others.

Isolation by pouring:

- Table 2
- Mix 1 was used for dilution of the pouring plates.
- E. coli was the only bacteria recognized in the pour plate.
- Absence of “sprinkles” on streak plates is the main difference when compared to the pour
plates.
- Microbes that were streaked on the surface of the agar plates grew more freely as those that
were mixed with the agar.

Calculated concentration:

- Plate 5 (from pour plates)

- 43 colonies (counted)
- 105 is the dilution factor
- 10 is the 0.1 ml sample plated

43 X 105 X10 = 43 000 000 bacteria/ml

Conclusion:

- Lab objective was met.

- Bacterial isolation was achieved; contributing to understanding of the principle
- Experiment helped recognize the differences between streak plates and pour plates
- Quantitative form was achieved by applying the formula:
o # of colonies on chosen plate multiplied by the dilution factor multiplied by 10
- Plate descriptions were capable due to the examples of morphology
- Control plates allowed for identification of type of bacteria in mixture.