MICROBIOLOGY 2132
LABORATORY SCHEDULE
SUMMER 2007
WEEK#
1
DAY
M
LAB#
1
EXERCISE
Ex. 3-1
DESCRIPTION
Introduction to Lab
-Go over safety, Lab rules and expectations of lab—CB
-Go over syllabus—TA
-Explain lab notebook, data sheets and lab write ups—TA
6/04/07
T
Introduction to Microscopy
-Prepared slides for initial use of scope
-Wet mounts with peppercorn and hay infusions
-Students need to become familiar with scope and how to use it
Microscopy Continued
-Continue with wet mounts if needed
2
6/05/07
Ex. 3-6
Staining
-Introduction use of control organisms—TA
-Demo station wil be set up for commercially prepared slides of capsule, spore
and flagella stains
W
3
Ex. 1-1
Ex. 1-2
6/06/07
Ex. 1-3
R
4
-Students will perform Gram’s stain from broth and slant/plate
-Students must correctly gram stain for credit for this lab. Time will be allowed
in follow labs to complete this
Medium Preparation
-Cover autoclaving and media preparation—TA
-Will have examples of plates, slants, stabs, broth
Aseptic technique
-Aseptic transfers of Gram positive and negative bacteria
(1) slant to broth, (2) slant to plate (3) plate to slant (4) broth to plate
-Streak plate for isolated colonies of both Gram positive and negative bacteria
supplied in lab
-Students must be able to streak plates for isolated colonies (at least 5 isolated
colonies per plate) for credit for this exercise. Time will be allowed in following
labs to complete this.
Staining
-Complete correct Gram’s stain if you have not already done so.
Results from Aseptic Technique
-Begin to talk about cultural characteristics—TA
6/07/07
-Students will check plates for at least 5 well isolated colonies
-Gram stain from 2 of the well isolated colonies for each type of bacteria and
look for contamination
-Gram stain broth and look for contamination
Staining
-Complete correct Gram’s stain if you have not already done so.
WEEK#
2
DAY
M
LAB#
5
EXERCISE
Ex. 2-1
6/11/07
Ex. 2-2
DESCRIPTION
Ubiquity of Microorganisms
-Students will collect 5 different environmental samples on TSA plates.
Incubate @ Room Temp(RT) 1-7 days
-Students will be allowed 20 minutes to collect these 5 samples in locations of
their choosing (inside and/or outside of LSE).
**Information for this investigation will be collected throughout the semester
and will be included in Lab Report II.**
*Students’ ubiquity organisms should be isolated and on slants by June
21*
Cultural Characteristics of Microorganism
-Go over cultural characteristic/colony morphology of organisms-TA
T
Staining
-Complete correct Gram’s stain if you have not already done so.
Begin Isolation/Identification of Bacterial Unknowns
-Students will be given cultures of mixed unknowns (cultures containing two
different organisms)
-Organisms will be mixed at the beginning of lab
-Students will streak two TSA plates for isolated colonies. Incubate @37C 24hrs
then move to RT until get good visible growth.
-Students will Gram stain their mixed culture
**Information from the Bacterial Unknown investigation will the basis for Lab
Report II—therefore keep good records of what you do and your
results.**
6
6/12/07
**Students’ mixed unknown organisms should be isolated and on
slants by June 21**
Ubiquity of Microorganisms Continued
-Students will check plates for growth (there may not be any on some plates as
some organisms grow faster than others)
-Examine and record any cultural and or plate characteristics for the Ubiquity of
Microorganisms Investigation. Share this information with class
-Streak for isolation 2 different colonies of interest from ubiquity samples
(remember not all will be able to do this at this until another lab)
-Check purity of colonies from ubiquity samples by performing Gram stain on
an isolated colony
W
7
Ex.2-10
Ex.2-11
Ex.2-12
6/13/07
Staining
-Complete correct Gram’s stain if you have not already done so.
Environmental Effects
-Effects of temperature, pH & osmotic pressure on microbial growth using
Nutrient broth and spectrophotometer to measure growth. Exercise will include
students ubiquity organism plus organisms provided in lab.
**The data from this Lab will be the basis for Lab Report I, which is
due on June 25, 2007**
Ubiquity of Microorganisms Continued
-Continue with ubiquity isolation for purity
-Once pure culture of ubiquity organism is achieved, subculture to 2 TSA slants
from streak plates to maintain throughout the rest of the semester.
*ubiquity culture must be maintained throughout the semester AND if not
viable at time of final exam points will be deducted*
Isolation/Identification of Bacterial Unknowns Continued
-Continue with mixed unknown isolation for purity
WEEK
DAY
R
LAB#
8
EXERCISE
DESCRIPTION
Environmental Effects Continued
-Measure OD of samples
-Interpret results by plotting OD vs temp, pH, osmotic pressure
Ubiquity of Microorganisms Continued
-Examine cultural characteristics of ubiquity samples and record information for
the Ubiquity of Microorganisms Investigation.
-Streak for isolation 2 different colonies of interest from ubiquity samples, if
you have not already done so
-Check purity of colonies from ubiquity samples by performing Gram stain on
an isolated colony
-Subculture putative pure colony to two slants or restreak a colony from plate
for isolation if necessary
6/14/07
Isolation/Identification of Bacterial Unknowns Continued
-Continue with mixed unknown isolation for purity
3
M
9
**Data sheets due for week of June 4**
Isolation/Identification of Bacterial Unknowns Continued
-Continue with mixed unknown isolation for purity
-Once pure cultures of mixed unknowns are achieved, subculture (per
organism) from streak plates to two TSA slants to maintain throughout the
rest of the semester.
6/18/07
**All students must have on TSA slants (1) two different pure
organisms from the mixed unknowns (2) a pure ubiquity organism
before they can use them in the exercises for Selective & Differential
Media and Biochemical test**
-Students will maintain their pure cultures on slants--two per organism
(commonly referred to as reserve slants) throughout the rest of the semester
*This is true for your organisms from the mixed unknown AND your ubiquity
organism*
Ubiquity of Microorganisms Continued
-Continue with ubiquity organism isolation for purity
Isolation/Identification of Bacterial Unknowns Continued
-Continue with mixed unknown isolation for purity
T
6/19/07
10
Staining
-Complete correct Gram’s stain if you have not already done so.
Ubiquity of Microorganisms Continued
-Continue with ubiquity organism isolation for purity
Isolation/Identification of Bacterial Unknowns Continued
-Continue with mixed unknown isolation for purity
**Organisms need to be streaked and isolated 3 separate times to
insure purity*
*Students should have completed at least one correct Gram’s stain by
this point. This technique will be part of the mid term practical*
WEEK
DAY
W
LAB#
11
EXERCISE
Ex. 7-3
Ex. 2-14
6/20/07
DESCRIPTION
Chemical Effects
-Antibiotic disk method on plates
-Also (1) Lysol (2)70% EtOH (3)10% Bleach using TSB broth and
spectrophotometers as in previous labs (9 & 10)
-Students will use their ubiquity organism plus organisms provided in lab
**Information from the Chemical Effects exercise will be the basis for Lab
Report 1**
Ubiquity of Microorganisms Continued
-Continue with ubiquity isolation for purity if necessary
R
Isolation/Identification of Bacterial Unknowns Continued
-Continue with mixed unknown isolation for purity if necessary
Chemical Effects Continued
-Measure OD of broth samples
-Interpret results by plotting OD vs Lysol, EtOH , Bleach
-Measure zones of inhibition on antibiotic disk plates
12
6/21/07
*Student need to have all 3 of their organisms isolated and on slants or
ready to be transferred to slants by this date*
**For the remainder of the semester, students will use streak plates
as the source of their organisms for use in exercises**
4
M
13
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
6/25/07
4-1
4-2
4-3
4-4
4-5
4-7
*Date sheets Due for week of June 11*
Selective & Differential Media
-Students will be introduced to media that selects for certain microorganism
and how some microorganisms can be differentiated--PEA, MacConkey, MSA,
EMB, etc. This will be approached as a continuation of inhibiting bacterial
growth rather than from a diagnostic view point—TA
-Students will be provided with organisms that react in various ways with the
media provided
**Students will use their ubiquity organism and purified mixed unknowns in
addition to organisms provided in lab.**
-TA’s this is an opportune time to reintroduce the practice of using control
organisms
**Lab Report I is due**
T
14
6/26/07
Selective & Differential Media Continued
-Students will continue with lab from previous day
-Students will check their media for growth (be sure and check lab manual for
incubation times) and gram stain organisms to confirm the selectivity of the
media
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organism
W
6/27/07
15
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
Selective & Differential Media Continued
-Students will check their media for growth and gram stain organisms to
confirm the selectivity of the media
Review For Lab Practical 1
WEEK
DAY
R
LAB#
16
EXERCISE
6/28/07
5
DESCRIPTION
Notebooks due
*Data sheets due for week of June 18*
M
17
7/02/07
T
18
7/03/07
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
Ex.
5-2
5-4
5-6
5-7
5-8
5-10
5-12
5-15
5-20
5-21
5-22
5-26
5-27
5-28
LAB PRATICAL 1
-Students will perform a Gram’s stain nd streak plate for grad
-Will be other stations set up related to material covered.
Biochemical Test for Gram positive and negative (Differential Tests)
-These tests are termed Biochemical tests because they differentiate organisms
based on specific biochemical characteristics.
-Tube media and Blood agar will used for these tests.
-Students will use their ubiquity organism and purified mixed unknowns in
addition to organisms provided in lab.
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organsim
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
Biochemical Test for Gram positive and negative (Differential Tests)
Continued From Previous Lab (#17)
-Continue with lab from previous day
-Students will look at results(be sure and check lab manual for incubation
times)
-TA’s will go over in detail the interpretation of each test result, including +/results and those results that do not fit into the biochemical chart—TA
**Students will use their ubiquity organism and purified mixed unknowns in
addition to organisms provided in lab.**
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organism
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
W
19
7/04/07
UNIVERSITY HOLIDAY-NO CLASS
R
20
7/05/07
Biochemical Test for Gram positive and negative (Differential Tests)
Continued From Previous Lab
-Students will look at results(be sure and check lab manual for incubation
times)
-TA’s will go over in detail the interpretation of each test result, including +/results and those results that do not fit into the biochemical chart—TA
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organism
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
*Data sheets due for week of June25*
6
7/09/07
M
21
Catch up time…report, data sheets, unknowns etc
WEEK
DAY
T
LAB#
22
EXERCISE
Ex. 8-4
READ
Ex. 6-1
7/10/07
DESCRIPTION
Serial dilutions and Plate counts
-Students will do pour plates and spread plates on soil samples they have
brought to class
-Students will do pour plates and spread plates on their ubiquity organism and i
incubate @RT 2-7 days
-Continue to work on identifying ubiquity and mixed unknowns
**Mixed unknowns and ubiquity organisms should be close to being
finished as report is due 7-19-07**
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organism
W
23
7/11/07
Ex. 8-2
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
Coliform-MPN-lab 1
-Students will use the MPN and perform the presumptive and confirmative test
for the presence of coliforms in an environmental sample
-Samples will be provided
-TA’s will remove samples after 48 hours and store @4C until next lab—TA
Serial dilutions and Plate Counts Continued
-Students will check samples
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organism
R
24
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
Serial dilutions and Plate Counts Continued
-Students will check samples
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organisms
7/12/07
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
7
7/16/07
M
25
Time for working on lab report II
Serial dilutions and Plate Counts Continued
-Students will calculate most countable plate and concentration
of organisms in original sample
-Compare pour and spread plate techniques
Coliform-MPN Continued (Lab2)
-Students will transfer all positive samples to BGLB and EC media
-TA’s will remove the media after 48 hours and hold until next
lab.
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organism
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
*Data sheets due for week of July 2*
WEEK
DAY
T
LAB#
26
EXERCISE
DESCRIPTION
Coliform-MPN Continued (Lab 3)
-Students will complete coliform lab
Serial dilutions and Plate Counts Continued
-Students will check samples if necessary
7/17/07
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organism
W
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
Ubiquity of Microorganisms Continued
-Continue with identification of ubiquity organism
27
7/18/07
Isolation/Identification of Bacterial Unknowns Continued
-Continue with identification of mixed unknowns
R
-Catch up time
Lab Report II is due
28
Ex.6.5
7/19/07
8
Growth Curve with Plate Counts
-Will use a fast growing organism that will allow several time points
from mid log to stationary phase to be collected
-Use OD readings and plate counts for plotting growth curve
29
*Data sheets due for week of July 9*
**Lab notebooks are due**
T
30
Growth Curve with Plate Counts Continued
-Students will interpret their data
-Students will perform plate counts
-Students will plot growth curve using regular graph and semilog
graph paper
-Students will calculate generation time
Finalize all labs
W
31
M
7/23-07
7/24/07
Lab clean up
7/25/07
Review for Lab Final
R
7/26/07
32
*Data sheets due for week of July 16*
LAB FINAL
Written only no practical applications.