Welcome to Implementing Inquirybased Microbial Project. Veronica Ardi, PhD

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1 Welcome to Implementing Inquirybased Microbial Project Veronica Ardi, PhD

2 Microbiology Laboratory Courses CourseSmart: ebook resources

3 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 Frustrating?

4 Microbiology Laboratory Courses M. Hos McGrane

5 Building on previous courses General Biology, Genetics, Molecular biology, etc. Scientific Method NHS

6 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:

7 Sample Timeline Does your order of techniques aid in inquirybased project? Johnson, T. and Case, C. Laboratory Experiments in Microbiology, 10 th edition, 2012; Pearson, Benjamin Cummings, San Francisco, CA

8 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 MIDTERM

9 Sample 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) Turn in Environmental Unknown Project

10 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: H 2 O 2 Coagulase test: rabbit plasma Gram stain reagents Glucose fermentation tube Nutrient agar plates & slants Personal protective equipment Sterile cotton swabs & sterile saline solution (or broth)

11 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 environment using cotton swab (dip in saline first).

12 Microorganisms in our world Go sampling! Any problems during sampling? Incubate Environmental samples NOAA OE

13 Results: variety of microbes

14 Results: Picking a colony Students would describe all the colonies from the sampling. Guide students to select 1 colony

15 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. aureus, etc.)

16 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)

17 Collecting Data

18 Collecting Data

19 Collecting Data

20 Collecting Data Enterotube II or API 20E APIWeb: https://apiweb.biomerieux.com/servlet/authenticate?action=preparelogin

21 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:

22 Analyses Identification of unknown applying Bergey s Manual Bergey s Manual: https://archive.org/details/bergeysmanualofd1957amer

23 Analyses Identification of unknown applying dichotomous key

24 Analyses Identification of unknown applying dichotomous key

25 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 Timing

26 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 Inadequate

27 Additional Assignments Paper: write a 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.

28 Top Student Interest Experiments that they have vested interest Experiments exploring themselves

29 Top Student Interest Experiments that they have vested interest Experiments exploring themselves

30 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. How can they apply it to a project?

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