Welcome to
Implementing
Inquiry-based Microbial Project
Microbiology Laboratory Courses
Microbiology Laboratory Courses
• Traditional Lab Manual• Individual experiments
• Defined results
• Post-lab questions with right answers
• Hypothesis-driven or Inquiry-based lab class
• Creative
• Critical thinking
Microbiology Laboratory Courses
Building on previous courses
• General Biology, Genetics, Molecular biology, etc.
• Scientific Method
Microbial Techniques
_____ Acid-fast stain
_____ Aseptic Technique _____ Biochemical tests _____ Capsule stain
_____ DNA isolation & sequencing _____ Gram stain _____ Growth curve / Spectrophotometer _____ Kirby-Bauer test _____ Microscope
_____ Most probable number
_____ Motility
_____ Negative stain _____ PCR
_____ Pure culture isolation _____ Serial Dilution _____ Selective Media _____ Simple stain _____ Spread plating _____ Streak plating _____ Wet mount _____ Other: ____________
Sample Timeline
• Does your order oftechniques aid in inquiry-based project?
• Johnson, T. and Case, C. Laboratory Experiments in Microbiology, 10 th
edition, 2012; Pearson, Benjamin Cummings, San Francisco, CA
Sample Timeline
Date/ 2015 Laboratory Topics General procedures, Lab Safety, Microscope Video, Turn in Safety Contract. Ex. 1: Parts of the Microscope & Oil immersion Ex. 3: Microbes in the Environment, Morphological unknown Ex. 26: Effectiveness of Hand washing Continue Ex. 3 & Isolate environmental sample Aseptic technique Ex. 11: Isolation of bacteria (& unknown environmental sample) Continue Ex. 11 & 26 (& unknown environmental sample) Ex. 5: Smears & Simple Stains (& unknown environmental sample) Ex. 7: Gram Staining (& unknown environmental sample) Ex. 12: Special Media (& unknown environmental sample) Ex. 20: Bacterial Growth Curve Continue Ex.12 Ex. 19: Oxygen & the growth of bacteria Lab MIDTERMSample Timeline
Date/ 2015
Laboratory Topics
Continue Ex.19
Ex. 14: Fermentation (& unknown environmental sample) Continue Ex. 14
Ex. 24: Disinfectants & Antiseptics Continue Ex. 24
Ex. 25: Antimicrobial drugs (& unknown environmental sample) Documentary: Nightmare Bacteria
Continue Ex. 25
Ex. 45: Bacteria of the Skin
Unknown environmental sample / project Continue Ex. 45
Ex. 46: Bacteria of the Respiratory Tract Unknown environmental sample / project Continue Ex. 46
Ex. 51: Enterotube ID unknown environmental sample Continue Ex. 51 Unknown environmental sample / project
Final Lab Exam (Cumulative)
General Equipment for Microbiology Lab
• Antibiotic disks (at least 5 different classes): ampicillin, penicillin, streptomycin, tetracycline, & vancomycin
[typically used in lab] or any that is typically used in clinics • BD Enterotubes II API 20E
• Blood agar, EMB, Mannitol salt agar (MSA), and Mueller-Hinton agar plates
• Bunsen burner Wire loops
• Catalase test: H2O2 Coagulase test: rabbit plasma • Gram stain reagents
• Glucose fermentation tube • Nutrient agar plates & slants • Personal protective equipment
Microorganisms in our world
• In pairs, come up with hypothesis or inquiry.
• How will you sample? [Design your method.]
• Procedure (Exp. 3): inoculate 1 plate from the
Microorganisms in our world
• Go sampling!• Any problems during sampling?
• Incubate
• Environmental samples
Results: Picking a colony
• Students would describe all the colonies from the sampling.
Alternatives in sampling
• If no growth or mostly fungus …• Instructors should complete an initial
environmental sample (i.e. sample the bottom of your shoes).
• Instructor’s can have pure cultures labeled as environmental unknown.
• Try not to use common bacteria (E. coli, S.
Timeline
Date/ 2015 Laboratory Topics General procedures, Lab Safety, Microscope Video, Turn in Safety Contract. Ex. 1: Parts of the Microscope & Oil immersion Ex. 3: Microbes in the Environment, Morphological unknown Ex. 26: Effectiveness of Hand washing Continue Ex. 3 & Isolate environmental sample Aseptic technique Ex. 11: Isolation of bacteria (& unknown environmental sample) Continue Ex. 11 & 26 (& unknown environmental sample)Isolate unknown environmental sample – pure culture
Ex. 5: Smears & Simple Stains (& unknown environmental sample) Ex. 7: Gram Staining (& unknown environmental sample)
Collecting Data
• Enterotube II or API 20E
Collecting Data
Identification of your unknown environmental sample:
If a patient had an infection from the environmental microbe that you identified above, then what antibiotic(s) could be used:
Analyses
• Identification of unknown – applying Bergey’s Manual
Bergey’s Manual:
https://archive.org/details/bergeysmanualofd1957amer
Analyses
• Identification of unknown – applying dichotomous key
Analyses
• Identification of unknown – applying dichotomous key
Actual Student Results
• What are the issues in identification of unknown?
• What happens when student’s are absent?
• What if there is no identification?
• Motivation
Rubric
• Establish a rubric to grade the project
• Unknown Environmental Project: 5-15% of the total grade
• Example of rubric found in packet Excellent Satisfactory Needs
Improvement
Additional Assignments
• Paper: write a 500-700 word paper about the bacterial organism was identified.
• Student presentation
• What should the paper / presentation include?
• A clinical case study, if applicable, should be required in the writing or presentation
assignment.
• Ecological significance of the bacterium should be part of the presentation or paper.
Top Student Interest
• Experiments that they have vested interest
Top Student Interest
• Experiments that they have vested interest
Limitations in Inquiry-based project
• This workshop is limited to bacterial samples.• What if the students do not understand the technique, how can they perform it?
• How can they interpret the test?
• Master the purpose of the test is key.
