Class Two Fall – Spring Semester

Exp 1: Culture Transfer Techniques, w/ organisms Materials: per table: cultures – broths for EC, SM,ML, and BS per person : media: 1 broth, 1slant,1 deeps, 1 plate ( min) Procedure: properly label each medium aseptically transfer, inoculate each organism to the three different media. Prep for incubation at 25C, /24hrs RF,BC, 1/27 BC SM RF,SM, 1/27

Organisms  SM, Serratia marcescens  ML, Micrococcus luteus  BS, Bacillus subtilis  EC, Echerichia coli

Comparison of E. coli and Micrococcus luteus

Journal Entry Exp 1 Culture Transfer Tech. date Materials: Cultures: BS, SM. Media:.. Equipment: … Procedures: bullet procedures, and ref page # in lab book ( citation: Cappucino & Sherman, 7 th ed. Pages : ---) Data: Conclusion: Pg # signe d

Culture characteristics  Use supplement and handout to observe the growth of the four organisms in the slant, deep, broth, and on the plate.  Do the organisms look like one of the examples on your sheet?  Try to record their appearance on your templates

Culture observations on the agar plate  Color production( chromogenesis). An example of this is the pink color of Serratia  Growth pattern and characteristics  Amount of growth( scant or heavy)

Colony morphology

Margin of the colonies

Elevation

Broth culture( refer to supplement)  Cloudy  Turbid( Flocculent)  Sediment formation  Pellicle formation

Slants  Is there growth in the bottom ?  Is there growth on the slant itself  What are the growth characteristics on the slant?  Key words Aerobic Anaerobic Facultative

Preparation  Label all tubes and plates carefully  Assign each member of the group 2 organisms  Transfer the organisms to the culture media using aseptic techniques used in weeks one and two

Organisms for study  Gram negative organisms  PA - Pseudomonas aeruginosa  PV- Proteus vulgaris  EC- Escherichia coli  EA- Enterobacter aerogenes  Gram Positive Organisms  BS - Bacillus subtilis  SA - Staphylococcus aureus  SE - Staphylococcus epidermidis  SS- Streptococcus salivarius

Journal Entry Exp 3. Exp 2A. Isolation of Pure Cultures date Purpose: In lab book Materials: Cultures: Mixed BS, SM. & SM, ML, Media:.. Equipment: … Procedures: bullet procedures, and ref page # in lab book ( citation: Cappucino & Sherman, 7 th ed. Pages : ---) Data: next period Conclusion: Pg # signed

Exp 2A, Isolation of Pure Cultures Streak Plate Dilution Technique, SPD Spread Plate Technique SM/ MLBS/SM cultures SP D SPD,SM/ML RFSPD,bs/sm RF SM/ML RFBs/sm RF Incubation, 25C, 24 hrs Materials: mixed cultures: SM/ML & BS/SM, one per table, 1 TSA Plates per person SM/ MLBS/SM cultures SP

Exp 2A, Isolation of Pure Cultures Streak Plate Dilution Technique, SPD Spread Plate Technique SM/ MLBS/SM cultures SP D SM/ ML cultures SP SPD,SM/ML RFSPD,BS/SM RF SM/ML RFBS/SM RF BS/SM Materials: mixed cultures: SM/ML & BS/SM, one per table, 4 TSA Plates per person 22C/24h r

Isolation of Pure culture  Observe your dilution streak of your mixed culture  On the bottom of your Petri dish circle colonies of two organisms  Example ML/SM mixture – circle yellow and pink cultures  With your inoculating loop lift cells from circled colonies and streak on new plate or inoculate a slant per detailed instructions in class