1. Microbiology Class Four Culture Characteristics

2. Day 4 Exp 2B, Isolation of pure cultures form SPD, Streak Plate Dilution, Technique. Procedure: page 15 & page 19 Exp 3, Culture Characteristics Discussion Procedure: page 23 Charts Case Study Review culture: (Exp 1, 2A) update Journals Quiz next Week

3. Class schedule( Part One)  Check results from class 3.  Record results in templates.  Organisms studied – class 3  S. marcescens  B. subtilis  M. luteus

4. 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

5. 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)

6. Comparison of E. coli and Micrococcus luteus

7. Colony morphology

8. Margin of the colonies

9. Elevation

10. Colony morphology

11. Broth culture( refer to supplement)  Cloudy  Turbid( Flocculent)  Sediment formation  Pellicle formation

12. 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

13. Part One – Completion of class 3 work  When you have finished observing all of your cultures  Place all tubes in rack in hazardous waste  Place all plates in cans in hazardous waste  Wipe down desk top

14. Ex# -2/B -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

15. Exp 3, Culture Characteristics Discussion Procedure: page 23 Exp 2B, Isolation of pure cultures form SPD, Streak Plate Dilution, Technique. Procedure: page 15 & page 19 Pick, using aseptic technique and a needle, a unique colony and transfer to a labeled slant. ML > yellow BS > white SM > red Culture per table: PA, EC,EA, PV, SA, SS, BS, SE, Inoculate each culture into a labeled media TSA plate, SPD TSA slant, surface TSA Broth Prep for Incubation @ Room Temp Prep for Incubation @ 37C Per table Day 4

16. Exp 2A, Isolation of Pure Cultures Streak Plate Dilution Technique, SPD Spread Plate Technique SM/ MLBS/SM cultures SPD SM/ ML cultures SP SPD,SM/ML RFSPD,BS/SM RF SM/ML RFBS/SM RF SM, Serratia marcescens,red ML, Micrococcus luteus,yellow BS, Bacillus subtilis, white BS/SM Materials: mixed cultures: SM/ML & BS/SM, one per table, 4 TSA Plates per person 22C/24hr review

17. New work( supply table) Experiment #3 Eight Organisms for Study/Table  8 Plates  8 Deeps( if available)  8 Slants  8 Broths

18. Key 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

19. 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

20. Review for Quiz One 1. Aseptic techniques 2. Culture transfer techniques 3. Media 4. Safety precautions 5. Microscope identification