Idea Transcript
MICROBIOLOGY LAB EXERCISES 15 WEEK SEMESTER Lab Session #
Topic and Page #
Supplies/ Cultures
1
Welcome to Lab Safety pg, ix – xi. View safety video
Safety Video and VCR or LCD
2 Brightfield Microscopy - pg. 3
3
Protozoans, algae, and Cyanobacteria - pg. 33
Total supplies & Culture for (1) one section (24) students
Some instructors might require prepared Blood slides. Slides of letter e. Instructors are to ensure that Students are to record results in form of drawings, etc and answer all questions at the end of each lab exercise during the entire semester.
Human Blood slides, slides with 3 bacteria morphologies; Prepared slides of Amoeba, Paramecium, Saccharomyces-yeast, Euglena, spirogyra
Commerical Protists (Euglena, Paramecium, diatoms, etc.)
Lots of covers slips
1 NA plates, 1 Blood agar plates per 3-4 students, 2 NB tubes per student, sterile cotton pads Students will ONLY use Exe. 37 as a comparison for colony growth and appearance
Wet mount techniques 4
Ubiquity of Bacteria pg. 47
NA plates, Blood agar plates, NB tubes, sterile cotton swabs/pads
5
Results from previous lab. Review Exe 37 pg. 245
Review Exe 37
6
Aseptic Technique - pg. 63
8 NB of E.coli 8 NA slant of M. smegmatis 8 NA plate with colonies (E.coli, M.luteus, S. marcescens)
Lab Aide should talk to the instructor about the number of media tubes and plates required by each student (24 each)
7
Record results from previous lab:
NA plate with colonies (E.coli, M. smegmatis, S. marcescens, M. luteus) form previous lab
8
Pure Culture techniques - pg. 73 Streak and Pour Plate Techniques
60 NA Deeps 125 sterile petri plates sterile 8 NB mixed culture and 8 NA plates of mixed cultures of S.marcescens, E.coli, M.luteus. Hot plates
9
Record results from previous lab. Continue with subculturing techniques
48 NA slants, three slant tubes per group of 3 students
10
Smear preparation and simple staining - pg.87 – 91
Clean glass slides, USE CULTURES FROM PREVIOUS LAB
11
Negative Staining – pg. 93 USE CULTURES FROM PREVIOUS LAB
NA plate with colonies (E.coli, M. smegmatis, S. marcescens, M. luteus) form previous lab
It is suggested that a negative control. Such as S. aureus be provided also.
12
Gram Stain - pg.101
8 NA plates of B.megaterium, 8 NA plates- S. aureus. Gram stain reagents (make sure iodine is the color of tea). Cultures should be no older than 24 hours.
For a Monday lab, it is suggested that cultures be set up on Friday and incubated at room temperature. Instructors should notify Lad Aid if they require additional/ different cultures.
8 NB - P.aeruginosa, 8 NB- M.smegmatis. Gram stain reagents (make sure iodine is the color of tea). Cultures should be no older than 24 hours.
For a Monday lab, it is suggested that cultures be set up on Friday and incubated at room temperature. Instructors should notify Lad Aid if they require additional/ different cultures.
Day 1, use cultures prepared on NA slants
13
Gram Stain - pg.101 Day 2, use cultures prepared on NA slants
Students should work in groups of 2, each will get 5 20 ml deeps and 5 NA plate
14
15
Spore Stain - pg. 105
Acid Fast Stain - pg. 103
8 NA slants or plates (24 -36 hours old) of B.megaterium, 8 NA plates of S.aureus hot plate, malachite green, safranin 8 NA slants or plates of M.smegmatis (48 hours cultures), 8 NB of S.aureus, hot plate, carbolfuchsin, acid-alcohol, methylene blue
It is suggested that a negative control. Such as S. aureus be provided also.
16
Motility Determination - pg. 107
8 NB of P.vulgaris 8 NB of Micrococcus luteus (cultures should be young)
2 tubes of SIM per 4 students, depression slides, clean slides, cover slip, Vaseline.
17
The Fungi - pg. 53
Prepared slides of S.cerevisiae* Broth cultures of S. cerevisiae, methylene blue stain, clean slides/cover slips
Students will also look at prepared mold slides that contain all three: Penicillium, Aspergillus, Rhizopus
18
Capsular Staining
capsular staining can also be done with Klebsialla pneumoniae
The cultures for these procedures are listed in the manual.
19
LAB Midterm Exam
20
The Standard Plate Count method of estimating bacteria population- pg. 139
8 NB E.coli, 5 - 20 ml NA deeps per 2 students, 5 sterile Petri plates per 2 students, 4 - 99 ml sterile water blanks per 2 students, 6 - 1.1 ml pipettes per student, biohazard bags, pipette pumps( 1 per group of 3 or 4)
Three 99ml sterile water blanks, 4 sterile empty petri plates and 4 sterile pipettes per group of 4 or 3 students. The Lab Aide should talk to instructor about the logistics of this exercise. How many blanks and plates per group?
21
Record results from previous lab.
Instructor should go over several quantitative plate count calculation problems with students.
22 Differential Plates Antimicrobic Sensitivity Testing –
The Lab Aide should prepare various media; i.e.; Na; EMB, MSA, MAC, CITRATE ACID, SIM etc…..
The Lab Aide should talk to instructor about the logistics of this exercise.
24 Plate MuellerHinton II agar 8 NB of each - S. aureus, E.coli, P. vulgaris, P. aeruginosa Cartridge of disk and disk dispenser
Antibiotic discs list: penicillin, streptomycin, erythromycin, kanamycin, ampicillin, tetracycline,
All students are to carry out the unknown identification exercise and submit a report on their investigation for grading
The Lab Aides should talk to each instructor about the logistics of this exercise. Each student/group will be given TWO organisms in their unknown cultures.
pg. 215
23 Continue Ex. 33 (Results observation and interpretation) 24 Start Unknown- pg.263
25 Continue Unknown
26 Continue Unknown
27 Continue Unknown
28
Final Exam (Clean up)
29 (Clean up)
30
*OPTIONAL LAB COULD NOT FIT THIS INTO PRESENT SCHEDULE
Cultivation of Anaerobes Lab exercise 19 in lab book
We could sub this lab in place of one the above labs, let’s discuss it ASAP.
Observe and record results from previous lab.
6 Fluid thioglycollate broth medium (FTM) tubes with resazurin dye per group of 3 students or per table. Gas Pack jar set- 1 per class 6 NA plates per group of 3 students or per table.
One 24 hr broth cultures of the following per group: M. smegamtis, S.aureus, M.luteus, E.coli, P. aeruginosa, C. rubrum