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Microbiology. Class Three Spring and Fall Semester Culture Characteristics. Day 3. 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.
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Microbiology Class Three Spring and Fall Semester Culture Characteristics
Day 3 • 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
Class schedule( Part One) • Check results from class 3. • Record results in templates. • Organisms studied – class 3 • S. marcescens • B. subtilis • M. luteus
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)
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
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
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
Day 4 • 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 Culture per table: PA, EC,EA , PV, SA, SS, BS, SE, Pick, using aseptic technique and a needle, a unique colony and transfer to a labeled slant. ML > yellow BS > white SM > red Inoculate each culture into a labeled media • TSA plate, SPD • TSA slant, surface • TSA Broth Per table Prep for Incubation @ Room Temp Prep for Incubation @ 37C
cultures cultures SM/ ML SM/ ML BS/SM SP SPD review Exp 2A, Isolation of Pure Cultures • Streak Plate Dilution Technique , SPD • Spread Plate Technique SM, Serratia marcescens,red ML, Micrococcus luteus,yellow BS, Bacillus subtilis, white BS/SM 22C/24hr SPD,SM/ML RF SPD,BS/SM RF SM/ML RF BS/SM RF Materials: mixed cultures: SM/ML & BS/SM , one per table, 4 TSA Plates per person
New work( supply table)Experiment #3 Eight Organisms for Study/Table • 8 Plates • 8 Deeps( if available) • 8 Slants • 8 Broths
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
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
Review for Quiz One • Aseptic techniques • Culture transfer techniques • Media • Safety precautions • Microscope identification