Saturday, July 7, 2012

Mr. Stowe's Microbiology Summer II class!

Class Syllabus
Lab Schedule
Lecture Schedule

Day 1 - Microscope
          - Mouth Sample

Day 2 - Aseptic Transfer Technique`
E Coli Broth and Slant

                                             S.M. Broth and Slant
Day 3 - Out for 4th of July

Day 4 - Simple & Gram stains

                                     Gram negative and Gram posotive stain mix -unsuccessful
                                                  Rods should be pink (they are purple)

Day 5 - Simple & Gram stains (contd)

Day 6 - Acid-Fast & Endospore stains

Acid-Fast Stain
(Mycobacterium Smegmatis)

Day 7 - Acid-Fast & Endospore stains contd.

Endospore Stain
(Bacillus Subtilis)
Day 8 - Lab Practical 1 Review
Day 9 - Lab Practical
Day 10 - No Class

Day 11 - Streak Plates

                                                                         SM
                                                                          EC

Day 12 - Selected Microbial Enzyme Assays


                                          Catalase Test - SE posotive for the catalase enzyme
                                        Oxidase Test - SA posotive for Cytochrome c oxidase
                   Nitrate Reduction Test - PA posotive reading after incubation (bubble in durham tube)
                                  EC posotive reading after adding reagent A and B (turned red)
                          Starch Hydrolysis - BS posotive for amylase enzyme (breaks down starch)
                                    Urea Hydrolysis - PV posotive for reducing urea to ammonia
                                    Coagulase Test - SA posotive (plasma has been coagulated)
                                                             Triple Sugar Iron Agar
                                          Salmanella - (black/yellow - H2S) sulfur reduction
                                          PA - (pink/no change - K/NC) no fermentation
                                          EC - (yellow/yellow - A/A) glucose & lactose fermentation

Day 13 - Carbohydrate Fermentation, IMViC Reactions, & OF-Glucose

                                   Methyl Red Test - EC posotive for mixed  acid fermentation   

                                               Citrate Test - EA posotive (citrate is utilized)

                                                      Oxidation Fermentation of Glucose
                                                    AF - entire tube green (non-reactive)
                                            PA - top only yellow (oxidizer - needs oxygen)
                                  EC - top and bottom yellow (fermenter - doesn't need oxygen)

Day 14 - Lab practical review
Day 15 - Lab practical

Day 16 - Selective and Differential Media

Motility Test

                                                 P. Vulgaris - posotive for motility
                                                      S. Epi - negative for motility

TSA Plain (mix)
                                                                 E.Coli and S. Sap.

TSA Sheep Blood

    E.F. -  partial hemolysis (alpha hemolysis)


PEA Agar

                                                   S.A. - growth and isolated colonies

Columbia CNA

                                                    E.F. - growth and isolated colonies

Mannitol Salt


S.E. - Pink Growth
S.A. - Neon yellow growth
S. Sap - Neon yellow growth

MacConkey Agar


                                                 E.F. - no growth (inhibited by chemical)
                                                   E.C. - hot pink (fermenter; posotive)
                                                           S. Ent - colorless growth
                                                       S.A. - nonfermenting (negative)

EMB Agar

                                                            E.A. - red pink (posotive)
                                                       E.C. - metallic green (posotive)
                                                               E.F. - clear (negative)
                                                         S. Ent - white pink (posotive)

Day 17 - Picked unknowns (#8), aseptically transferred to streak plates for isolation, transferred to slides to do gram stains to check for pink and purple for negative and posotive

                                                              Gram negative Bacilli

                                                                 Gram posotive Cocci
                                                     Gram negative unknown bacteria

                                                      Gram posotive unknown bacteria

Day 18 - Tests performed on both gram negative and gram posotive unknowns

-Catalase test performed on gram posotive bacteria - posotive result (bubbles)

-OF Glucose for gram negative bacteria
                                                                        Fermenter

Day 19 - More Tests

-Coagulase test for gram posotive bacteria
                                                          Negative result (liquid)
- MacConkey for gram negative bacteria - growth
- Oxidase for gram negative bacteria - negative result (no color change)

Day 20 - More Tests

- Lactose PR Broth for gram negative bacteria - posotive result
- Glucose PR Broth for gram posotive bacteria - posotive result

- Mannitol Salt for gram posotive bacteria - negative result (no growth)
- Nitrate test for gram posotive bacteria- negative result (turned red after zinc was added)
- Citrate test for gram posotive bacteria - posotive result (turned blue on top) = Kocuria Rhizophila!!!!

- VP test for gram negative bacteria - negative result
- Citrate test for gram negative bacteria - negative result (no color change) = Escherichia Coli!!!!

PHONE LOST ALL OF MY PICTURES SO I WASN'T ABLE TO UPLOAD ANYMORE! SORRY!