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 Mix 2 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) Serratia marcescens (S.M)

TABLE 2 - Isolation by Pouring Results/Discussion PLATE 3 Mix1 diluted X 103 4 Mix1 diluted X 104 5 Mix1 diluted X 105 6 Mix1 diluted X 106 7 Mix1 diluted X 107 DESCRIPTION/OBSERVATION 'sprinkles', blotches, round colonies, many growing into the agar Growing into agar, irregular, round, sprinkles Growing into agar, irregular, round, sprinkles Relatively little growth, growing into agar , sprinkles Irregular, spreading, moderate growth, growing into agar, sprinkles

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.

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