Welcome to BIO 205 Laboratory

1
Welcome to BIO 205 Laboratory!

• Section E
• Class Number 3269
• Tuesday 600-900

2
What are we doing today?

• Introduction to Microbiology Lab
• Lab Manual
• Laboratory Safety
• Assign Mini Writing Assignment 1
• Begin Environmental Isolate Project

3
Contact Information

• My name Camille Naaktgeboren
• Email cen28_at_nau.edu
• Phone [phone] (on campus 3-1373)
• Office Room 112, building 21, (right around
  the corner)
• Laboratory Manager Linda Fitchett
• Email Linda.Fitchett_at_nau.edu
• Phone [phone] (on campus 3-7239)
• Office Room 114 building 21, (across the
  hall)
  
• My office/open lab hours 500-600 Tuesdays,
  400-500 Wednesdays
• there are other open hours with different
  instructors, the schedules are posted outside
  this door and the door of room 125

4
Phantom students

• Students who have taken the lecture and lab
  previously, are re-taking the lecture portion of
  the course and want to transfer the previous lab
  grade into the lecture
• If this is you, fill out a 3x5 card with name,
  student ID , email address, the semester and
  section you took it last time, the grade you
  think you had, and a phone number if you want to
  (not required due to privacy restrictions)

5
Lab Manual and Required Texts

• Manual can be purchased at Kinkos/FedEx in
  Safeway Plaza
• Two other texts at bookstore
• Titles of those are in syllabus that is in lab
  manual
• Syllabus-read over this week, discussion of
  pertinent info. next week

6
Other Required Materials

• Lab coat
• can be bought at the bookstore
• May be able to find a button down shirt at a
  thrift store but must reach at least to your
  knees
• Fine tipped Sharpie

7
Mini Writing Assignment 1

• Due next week!
• Objective Reading and following directions
  thinking about the importance of microbiology
• Found in your lab manual after syllabus and
  rubrics

8
Lab Protocol and Safety

• List of Laboratory Rules in lab manual, page vii
• Page viii-ix, more detailed explanation of
  particular activities
• Provided supplies
• Safety quiz-when we come back from Environmental
  Isolate/break

9
Environmental Isolate Project

• This is a semester long project
• You will have to work on it outside of class
  times
• Today
• Collect microorganisms from the environment
• Throughout the semester
• Isolate and characterize one species

10
Begin Environmental Isolate Project

• Demonstration
• Label TSA plates name, date, lab section,
  exercise
• Aseptic handling of sterile swabs and plates
• Streak plates
• Control plate
• A control is a reference point in an experiment
  in which the set of conditions does not vary
• Collect microorganisms
• Inoculated TSA plates
• Incubate upside down at room temperature

11
Environmental Isolate

• Obtain Sterile Cotton Swabs and three TSA plates
• One plate is a control, the remaining two plates
  will be used for your organisms.
• As a group, we will go outside, swab, and streak
  material onto the TSA plates as demonstrated in
  class.
• No material that you suspect would have been in
  contact with a human source

12
Safety Quiz

• 1. When coming in to check cultures, you DO NOT
  have to wear your lab coat.
• TRUE FALSE
• You must ALWAYS wear your lab coat when in
  the lab.
• 2. Eating, drinking and applying lip-gloss while
  in the laboratory IS PERMITTED.
• TRUE FALSE
• Eating, drinking, and applying lip-gloss is
  NEVER permitted in the laboratory.
• 3. If your microscope is NOT in good working
  condition, you should try and fix it yourself.
• TRUE FALSE
• Bring microscope problems to the attention
  of your TA.

13
Safety Quiz

• 4. You should assume that the inoculating
  loop/needle is contaminated until it is
  sterilized in the flame.
• TRUE FALSE
• 5. If an accident occurs where culture is
  spilled, you should clean it up immediately.
• TRUE FALSE
• Notify TA immediately of any culture spills.
• 6. When using culture tubes, it is acceptable to
  place them in the student bins with the stains
• TRUE FALSE
• ALWAYS place culture tubes in racks.

14
Safety Quiz

• 7. Under certain circumstances, students may
  remove cultures from the laboratory.
• TRUE FALSE
• NEVER remove cultures from the laboratory.
• 8. You must wash your hands with soap and water
  upon arriving and before leaving the laboratory.
• TRUE FALSE
• 9. You must wash you work bench with bleach and
  a damp sponge upon arriving and before leaving
  the laboratory.
• TRUE FALSE

15
Safety Quiz

• 10. It is NOT necessary to wear safety glasses
  when working with bacterial cultures.
• TRUE FALSE
• You are to wear safety glasses whenever
  you are working with bacterial cultures.
• 11. Gloves MUST be worn when working with stains
  and reagents that may be toxic to the skin.
• TRUE FALSE
• 12. If you are pregnant, or otherwise
  immunocompromised (i.e. HIV/AIDS, chemotherapy
  treatment, etc.), you may be at greater risk than
  otherwise.
• TRUE FALSE

16
Safety Quiz

• 13. Each student is held responsible for the
  equipment that is either assigned to or used by
  them.
• TRUE FALSE
• 14. If a culture tube is dropped or broken, the
  student should clean it up immediately and then
  notify the TA.
• TRUE FALSE
• Immediately notify your TA if culture
  tubes are spilled, dropped, or broken.
• 15. Broken glass, such as slides or cover slips,
  should be disposed of into a waste basket or the
  Nalgene buckets provided on the lab bench.
• TRUE FALSE
• Slides and cover slips are ALWAYS placed
  in slide trays at the ends of the benches.

17
Safety Quiz

• 16. Open wounds should be covered before coming
  into the lab.
• TRUE FALSE
• 17. Mouth pipetting is allowed only if the TA is
  assisting you.
• TRUE FALSE
• You should NEVER mouth pipette.
• 18. Your lab coat should be laundered and
  bleached periodically during the semester.
• TRUE FALSE

18
Safety Quiz

• 19. Open-toed shoes are permitted in the
  laboratory during summer sessions only.
• TRUE FALSE
• Open-toed shoes are NEVER permitted in
  the laboratory.
• 20. Failure to follow directions from the
  laboratory instructor may result in dismissal
  from the class (both lecture and lab).
• TRUE FALSE

19
Environmental Isolate Project

• Medium
• Liquid or solid preparation made for the growth
  and storage of microorganisms or other types of
  cells
• Can be used for broad range of microbes or very
  specific ones
• Trypticase Soy Agar Plate (or TSA Plate)
• Medium used today for collection and growth of
  microorganisms from the environment

20
Environmental Isolate Project

• Agar
• Widely used as a base for solid and semi-solid
  media
• A transparent gelatin-like substance formed by
  heating agar and water over 100C then cooling
• Rhodophyceae algae (red-brown algae) is a primary
  source of agar

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Environmental Isolate Project

• Why use agar?
• Agar gels at moderate temperatures (40-45C)
• Once set, agar is stable at temperatures up to
  65C or higher
• Few organisms are able to degrade agar

22
Environmental Isolate Project

• Growth media are prepared by adding nutrients,
  selective agents, etc. to agar
• Nutrients added to agar to make TSA Plates
• Digests of casein and soybean meal
• NaCl
• Agar plates dehydrate rapidly
• Always store plates upside down

23
To Do For Next Week

• Mini writing assignment 1
• Read exercise 1
• Get all required materials if you have not done
  so already