READY! 4/9/20 LIBRARYASSIGNMENT BONUS: (with NEW bacteria 4 April 2020) (25 all class BONUS POINTS)
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YOU are given this assignment without much explanation in order to assess your ability to read and follow directions. A short introduction may or may not be given in class... PLEASE READ THE ENTIRE PAGE BELOW!!! DO NOT SHOWER ME WITH EMAILS & QUESTIONS - learn to follow the directions!
This assignment if completed and submitted ON TIME counts 1% of your class final grade as part of the 3-4% CLASS WIDE BONUS POINTS - AND REQUIRED of ALL STUDENTS.
If you do this EARLY it will help you do well on the MIDTERM EXAM & Unknown Paper!
To do this assignment you will need to...
*WHAT TO TURN-in FROM your Group?
A Paper with a cover sheet listing the people in your group that actually worked on the project (last name first). A statement on the cover page saying that you each worked together and contributed to the answers.
I WANT 2 printed TABLES (you may hand write into the blank charts) and then scan or Microsoft LENS it to me at hicksmicro@gmail.com
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And TWO SENTENCES submitted to me as..."Unknown "A" is ________ (Genus + species) and "Unknown "B" is ________ (Genus + species).
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INDIVIDUALLY go to CANVAS QUIZZES TO GET YOUR CREDIT FOR YOUR UNKNOWN ''A'' & "B" AND ANSWER THESE QUESTIONS:
Answer these questions by listing each test:
1) Which of our tests measures use of GLUCOSE?
2) Which of our tests measures use of LACTOSE?
3) Which of our tests TELL YOU GRAM NEGATIVE OR POSITIVE?
4) Which of our tests measures use of CITRATE?
5) Which of our tests indicates ALCOHOLIC FERMENTATION?
6) WHAT is REVERSION?
7) Which of our tests measures use of CASASE?
8) Which of our tests measures THE PRODUCTION OF HYDROGEN SULFIDE?
9) Which of our tests IS THE WORST FOR LACTOSE USE?
You are to work in assigned groups of FOUR (4)... ALL names on one submission. Everyone will receive about 25 points for the Library Assignment (depending on the term & typed) if you did the PROCESS correctly and didn't just put "stuff." Making up data is cheating! These points will be ADDED to your classwide BONUS points in the class. This assignment is REQUIRED
PUT ALL OF YOUR NAMES (Last, First and your class + date) on your paper to turn in the UPPER RIGHT-HAND CORNER of the first page. Your total "over-the-top" BONUS points cannot exceed 3-4% of the Total Class Grade - no matter how many are given for the Cards and Library Assignment!
A Paper with a cover sheet listing the people in your group that actually worked on the project (last name first). A statement on the cover page saying that you each worked together and contributed to the answers.
I WANT 2 printed TABLES (you may hand write into the blank charts) and then scan or Microsoft LENS it to me at hicksmicro@gmail.com
&
And TWO SENTENCES submitted to me as..."Unknown "A" is ________ (Genus + species) and "Unknown "B" is ________ (Genus + species).
--------------------------------------------------------------------------------
INDIVIDUALLY go to CANVAS QUIZZES TO GET YOUR CREDIT FOR YOUR UNKNOWN ''A'' & "B" AND ANSWER THESE QUESTIONS:
Answer these questions by listing each test:
1) Which of our tests measures use of GLUCOSE?
2) Which of our tests measures use of LACTOSE?
3) Which of our tests TELL YOU GRAM NEGATIVE OR POSITIVE?
4) Which of our tests measures use of CITRATE?
5) Which of our tests indicates ALCOHOLIC FERMENTATION?
6) WHAT is REVERSION?
7) Which of our tests measures use of CASASE?
8) Which of our tests measures THE PRODUCTION OF HYDROGEN SULFIDE?
9) Which of our tests IS THE WORST FOR LACTOSE USE?
You are to work in assigned groups of FOUR (4)... ALL names on one submission. Everyone will receive about 25 points for the Library Assignment (depending on the term & typed) if you did the PROCESS correctly and didn't just put "stuff." Making up data is cheating! These points will be ADDED to your classwide BONUS points in the class. This assignment is REQUIRED
PUT ALL OF YOUR NAMES (Last, First and your class + date) on your paper to turn in the UPPER RIGHT-HAND CORNER of the first page. Your total "over-the-top" BONUS points cannot exceed 3-4% of the Total Class Grade - no matter how many are given for the Cards and Library Assignment!
GUIDELINES to help you:to determine the GENUS for Unknowns A and B... Follow the Gram Negative or Positive "keys" and determine the unknown Genus from the chart I supply (GENUS A and B)... COPY the info for 3 the microbes from the 7th Ed. of Bergey's IT IS ONLINE click here... and use it to...
- Submit the 2 charts plus 2 sentences stating the name for the Genus A + species & Genus B + species THIS is usually due BEFORE or on the MIDTERM Day and before your Unknown paper in order to receive Bonus points.
- DO NOT USE other dichotomous keys (only the keys supplied for this assignment will give you the correct answer).
- DO NOT USE ANY REFERENCE EXCEPT the 8th Ed. of Bergey's Determinative!
Part of this assignment assumes that you know our Unknown Tests from your Laboratory Guide and that the Bergey's Manual that you are using does not have a results chart.... You must learn to prepare such a chart of results to use with your Genus key... ALL the LAB TESTS will be on Exam 2. Have you looked at the old TESTS? DUMB MOVE IF YOU DON'T!!!
NOTE: 1 copy of the Blank Data Page is used as your unknown microbe's data... You may use my blank data chart to record YOUR unknown results... Staple this Data Page into the LAB SECTION of your Lab Record Book. Please note that when you write your paper the TESTS list should be alphabetized...
YOU HAVE 3 Assignments to Complete (USING Bergey’s Determinative 7th Ed ONLY):
(1) You are assigned to go to a Library and obtain a copy of the information on the following Genus and species from the 7th Edition of Bergey's Manual of Determinative Bacteriology.. JUST COPY THE PAGES THAT APPLY TO EACH MICROBE - PLEASE DO NOT TEAR-OUT ANY PAGES or ATTEMPT TO "check-out" the Bergey's Manual (consideration of others?). This assignment is due directly AFTER Spring Break or the class day BEFORE MIDTERM EXAM - which is the beginning of the Unknown Isolation...
COPY BOTH THE GENUS information and the species information for each microbe. If the Genus info contradicts the species - the species wins!
(a) Haemophilus influenzae (copy this in the 7th Ed. so you know how I did the chart below) DO THIS ONE FIRST!
(b) PROTEUS mirabalis
(c) Shigella dysenteriae
(alphabetize the tests please)
HINTS TO UNKNOWN WORK & UNKNOWN A/B
Library Assignment Bonus Page PDF (alphabetized)
* = These fermentation results are a part of the Enterotube II or Oxiferm Tube Tests only... see those tests for information
** = These fermentation tests are done as a part of the Enterotube II or Oxiferm Tube Test AND other tests that we do such as the PR Broths, etc.
*** = These are Agar Plate Tests that require melting, pouring and cooling a petri dish of Nutrient or TSA agar deeps and then you must use a sterile "Q-tip" to place a "LAWN" of your unknown on the agar BEFORE you add the discs or UV Light... lastly you must incubate at Opt. Temperature overnight and read or refrigerate the following day... SEE DISC TESTS online
NOTE: Anything called a "Plate Test" is an agar deep that is melted and poured into a petri dish, loop-line inoculated and incubated overnight at Opt. Temperature... Look at the bottom of the Agar Plate Test page on the website to see how to do all of the individual Agar Plate Tests... SOMETIMES what grows on a plate (selective media) or DOESN'T GROW is IMPORTANT INFORMATION!
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THE GENUS AND SPECIES of UNKNOWN A is:____________________________.
What is Unknown #A GENUS:________________ 10 pts ____________ species? ( 4 more Pts)
UNKNOWN MICROBE "B" RESULTS CHART: NOTE: "blank" means - no information given
What is Unknown #B GENUS:________________ 10 pts ____________ species? ( 3 more Pts)
BLANK DATA TABLE (MAKE YOURSELF 4 COPIES - Using the PDF link Above)… 1 for P. citreus, 1 for C. bovis, 1 for H. vaginalis and 1 for your personal Unknown from class… The left column is “every” test we could run in Microbiology at this college… the right column is for your results… If you cannot find the info - leave it BLANK! There are many "blanks."
PROPERTY
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POSITIVE or NEGATIVE (DESCRIPTION)
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First named by
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Discoverer
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| ISOLATION Procedure? |
(NOT USUALLY GIVEN IN A TEXT but useful for you in your unknown)
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| Motility | |
Thioglycollate Test (also called Oxygen Determination T)est
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Optimum Temperature Procedure
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Shape by observation
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Capsule Stain
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| Gram Stain | |
| Acid Fast Stain | |
Endospore Stain
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Growth on agar
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Colony Morphology
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Methyl Red Test
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Vogues Prosk. (VP)
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Starch (Amylase) Agar Plate Test
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FERMENTATION TESTS:
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Glucose**
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Glucose (anaerobic)*
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Maltose*
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Arabinose*
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Fructose*
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Xylose*
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Rhamnose*
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Galactose*
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Lysine*
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Sucrose**
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Dulcitol**
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Inositol*
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Lactose**
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Mannitol**
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Sorbitol**
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Ornithine*
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Adonitol*
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Saccharose**
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Arginine*
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OTHER TESTS:XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX
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RESULTS:
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Oxidase Test (Using old slant)
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Nitrate Broth Test
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Casease (2 - Skim Milk Agar Plate MAIN also secondary from Litmus Milk Test )
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Phenylalanine Deaminase
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Lipase (2 - Spirit Blue Agar Plate &/or Tributyrin Agar Plate Test)
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Oxygen Utilization (see Thio Broth above)
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Citrate utilization (2 - Simmon's Citrate Agar Slant/Deep &/or Koser's Citrate Broth Test)
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Salt agar growth 5% or 10% NaCl or Mannitol Salt Agar
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Catalase Test (using growth on a clean slide)
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Tellurite Glycine Agar Plate Test
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Hydrogen Sulfide Production (6 or The Iron Tests: Lysine Iron, Peptone Iron, Triple Sugar iron, Kliger's Iron &/or the Hektoen Agar Plate Test)
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Indole (Tryptophane Broth Test)
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Urea Broth or Urea Agar Test
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Gelatin Test
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ONPG test
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Litmus Milk Test
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Kirby Antibiotic Sensitivity Test***: sensitive to...
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Enterotube II Test for Gm- oxidase - rods only
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Oxiferm Tube Test for Gm- oxidase + rods only
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Antibiotic Disc Test (also known as the Kirby Disc Test)***
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Antiseptic/Disinfectant Disc Test***
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UV Light Sensitivity Test***
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(2) Make a chart for each organism from the reading... using ONLY the items we measure in OUR UNKNOWN TESTING! Also, remember to include the written information found UNDER THE GENUS in the Bergey's... It is best to make a copy of the information on the Genus and species for each microbe for each person in your group and then "DISCUSS" what should be recorded as a STUDY GROUP!
I designed the chart below as an example: Haemophilus influenzae
Here is my chart from the same text concerning the microbe: compare mine to the chart you made from your readings of the Bergey's 7th Ed.
PROPERTY:XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX
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POSITIVE or NEGATIVE (DESCRIPTION):XXXXXXXXXXXXXXX
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First named Lehmann & Neumann 1896
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Discoverer
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3 Main Isolation Techniques: (a) Spread Plate (b) Pour Plate (c) Streak Plate
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Streak plate isolation
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Motility (2 - Motility Gel Stab &/or Motility Hanging Drop Technique)
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negative
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Thioglycollate Test (also called Oxygen Determination T)est
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aerobic/facultative anaerobe
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Optimum Temperature Procedure
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37 C
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Shape by observation
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very small rod singles & pairs
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Capsule Stain
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sometimes seen
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Gram Stain
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Gram negative
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Acid Fast Stain
| |
| Endospore Stain | |
Growth on agar
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blood agar is best
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Colony Morphology
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small circular transparent - entire
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Methyl Red Test
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positive
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Vogues Prosk. (VP)
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negative
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Starch (Amylase) Agar Plate Test
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positive
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FERMENTATION TESTS:
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FEMENTATION RESULTS:
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Glucose**
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some do
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Glucose (anaerobic)*
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(***leave blank if no info)
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Maltose*
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Arabinose*
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Fructose*
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Xylose*
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Rhamnose*
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Galactose*
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Lysine*
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Sucrose**
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Dulcitol**
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Inositol*
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Lactose**
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no fermentation
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Mannitol**
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no fermentation
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Sorbitol**
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Ornithine*
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Adonitol*
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Saccharose**
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Arginine*
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OTHER TESTS:XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX
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RESULTS:
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Oxidase Test (Using old slant)
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positive
|
Nitrate Broth Test
|
positive
|
Casease (2 - Skim Milk Agar Plate MAIN also secondary from Litmus Milk Test )
|
negative
|
Phenylalanine Deaminase
| |
Lipase (2 - Spirit Blue Agar Plate &/or Tributyrin Agar Plate Test)
| |
Oxygen Utilization (see Thio Broth above)
|
Aerobic to Facultative
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Citrate utilization (2 - Simmon's Citrate Agar Slant/Deep &/or Koser's Citrate Broth Test)
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Salt agar growth 5% or 10% NaCl or Mannitol Salt Agar
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no growth on 0.5% NaCl
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Catalase Test (using growth on a clean slide)
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positive
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Hydrogen Sulfide Production (6 or The Iron Tests: Lysine Iron, Peptone Iron, Triple Sugar iron, Kligler's Iron &/or the Hektoen Agar Plate Test)
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Indole (Tryptophan Broth Test)
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usually positive
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Urea Broth or Urea Agar Test
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Gelatin Test
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negative
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ONPG test
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Litmus Milk Test
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ALK or purple/NO CASEASE no clearing
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Kirby Antibiotic Sensitivity Test***: sensitive to...
| |
Enterotube II Test for Gm- oxidase - rods only
| |
Oxiferm Tube Test for Gm- oxidase + rods only
| |
Antibiotic Disc Test (also known as the Kirby Disc Test)***
| |
Antiseptic/Disinfectant Disc Test***
| |
UV Light Sensitivity Test***
|
** = These fermentation tests are done as a part of the Enterotube II or Oxiferm Tube Test AND other tests that we do such as the PR Broths, etc.
*** = These are Agar Plate Tests that require melting, pouring and cooling a petri dish of Nutrient or TSA agar deeps and then you must use a sterile "Q-tip" to place a "LAWN" of your unknown on the agar BEFORE you add the discs or UV Light... lastly you must incubate at Opt. Temperature overnight and read or refrigerate the following day... SEE DISC TESTS online
NOTE: Anything called a "Plate Test" is an agar deep that is melted and poured into a petri dish, loop-line inoculated and incubated overnight at Opt. Temperature... Look at the bottom of the Agar Plate Test page on the website to see how to do all of the individual Agar Plate Tests... SOMETIMES what grows on a plate (selective media) or DOESN'T GROW is IMPORTANT INFORMATION!
=============================================================================
(3) Take the following test results information, make a Results Chart for Unknown A and B. Click on the "Genus Keys" below and determine the most likely GENUS for this UNKNOWN "A and B" then go to the Library and look up that Genus and determine your species... (Bergey's Determinative, 8th Ed). Remember to pick your species avoid pathogens of man and look first at the first on the list or TYPE species.
DATA: (Changed each term)
UNKNOWN MICROBE "A" RESULTS CHART: NOTE: "blank" means - no information given
PROPERTY Unknown #A
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POSITIVE or NEGATIVE (DESCRIPTION)
|
First named by
| ? |
Isolation technique
| Quadrant streak from feces of new borne |
Motility
| fuzzy pink throughout the tube |
Thioglycolate Test
| cloudiness at the bottom |
Optimum temperature
| 37 |
Shape
| bacillus |
Capsule Stain
| black |
| Flagella stain | flagella |
Gram Stain
| ooops broke the slide |
Acid Fast Stain
| blue after staining |
| Endospore Stain | pink with holes after staining |
Growth on agar
| lobate; very small, flat |
Colony Morphology
| see above |
Methyl Red Test
| |
Voges-Proskauer. (VP)
| |
Starch (Amylase) Test
| |
| Hektoen agar / MacConkey's agar / EMB agar | didn't grow on all 3 |
| KIA | yellow in bottom areas pink on top no black, cracks in all spots in agar |
| Lysine Iron | no black with purple slant yellow butt |
FERMENTATION TESTS:
| SEE BELOW! |
Glucose
| day 3 pink; day 5 the tube is layered orangey-yellow but yellow after mixing with a 1/2 inch bubble in Durham; |
Glucose (anaerobic)
| not tested |
Maltose
| pink with gas |
Arabinose
| PR broth day 5 = orange and no bubble |
Fructose
| yellow with 1/2 in bubble at day 5 |
Xylose
| 24 hrs yellow with gas |
Rhamnose
| pink broth 7 days no bubble |
Galactose
| pink/white broth 7 days bubble |
Sucrose
| 7 days pink/white with gas |
Dulcitol
| pink broth 7 days no bubble |
Inositol
| pink broth 7 days no bubble |
Lactose
| pink/white with gas |
Mannitol
| day 3 yellow with gas |
Sorbitol
| pink-yellow-orange broth 5 days no bubble |
Ornithine
| day 2 orange |
Adonitol
| pink broth 7 days no bubble |
Saccharose
| pink broth 7 days no bubble |
Arginine
| pink broth 7 days no bubble |
OTHER TESTS:
| SEE BELOW! |
Oxidase Test
| |
Nitrate
| on day 7 no change after reagent A + B then +Zn and mixing 4 min very faintly pink |
Casease
| |
Phenylalanine Deaminase
| |
Lipase
| |
Oxygen Utilization (see Thioglycolate Broth)
| see above |
| nothing not even a wisp up from the bottom after thumping 7 days; Simmon's green | |
Salt agar growth
| No growth |
Catalase Test
| looks dissolved |
Hydrogen Sulfide Production (Klig.& Peptone Iron Tests)
| yellow on bottom pink on top cracks at 3 days |
Tryptophane Test (Indole)
| |
Urea Test
| yellow |
Gelatin Test
| sliquid just out of incubator solid at 30 min at 5C after 21 days |
ONPG test
| white |
Litmus Milk
| blue/puple and cloudy |
Kirby Antibiotic Sensitivity Test: sensitive to...
| not done |
Enterotube II Test for Gm- oxidase - rods only
| not done |
Oxiferm Tube Test for Gm- oxidase + rods only
| not done |
Antibiotic Disc Test
| not done |
Antiseptic/Disinfectant Disc Test
| not done |
UV Light Sensitivity Test
| not done |
Tellurite Glycine Agar
| not done |
What is Unknown #A GENUS:________________ 10 pts ____________ species? ( 4 more Pts)
UNKNOWN MICROBE "B" RESULTS CHART: NOTE: "blank" means - no information given
IVE (DESCRIPTION)
| |
First named by
| ? |
Isolation technique
| Quadrant streak from feces |
Motility
| broke the tube but 7 min hanging drop vibrated under 100X |
Thioglycolate Test
| cloudy throughout tube |
Optimum temperature
| 37 |
Shape
| coccobacillus |
Capsule Stain
| black |
| Flagella stain | peritrichous by staining |
Gram Stain
| ooops broke the slide |
Acid Fast Stain
| blue after staining |
| Endospore Stain | pink no holes after staining |
Growth on agar
| entire, moist, grey |
Colony Morphology
| entire, moist, grey |
Methyl Red Test
| orangy red after mixing |
Voges-Proskauer. (VP)
| brownish green immediately after reagents same next day sit overnight |
Starch (Amylase) Test
| brown after iodine |
| Hektoen agar / MacConkey's agar / EMB agar | grew clear on all, salmond/hot pink on orange, greenish on purple agar |
| KIA | flecks of black |
| Lysine Iron | no black |
FERMENTATION TESTS:
| SEE BELOW! |
Glucose
| yellow+ bubble |
Glucose (anaerobic)
| not tested |
Maltose
| yellow + gas |
Arabinose
| acid+bubble |
Fructose
| acid+bubble |
Xylose
| acid+bubble |
Rhamnose
| yellow+bubble at 24 hrs |
Galactose
| acid+bubble |
Sucrose
| red after 5 days |
Dulcitol
| red after 7 days |
Inositol
| not done |
Lactose
| not fast after day 4 yellowish+bubble |
Mannitol
| acid+bubble |
Sorbitol
| broke tube |
Ornithine
| missing in incubator |
Adonitol
| pink after 7 days |
Saccharose
| not done |
Arginine
| not done |
OTHER TESTS:
| SEE BELOW! |
Oxidase Test
| black, faintly pink for 2 seconds then grey |
Nitrate
| reagent A +B+mixing=pink no Zn needed |
Casease
| has whey/clearing in Milk agar |
Phenylalanine Deaminase
| no change |
Lipase
| no change |
Oxygen Utilization (see Thioglycolate Broth)
| cloudy through slightly more at the top |
| Litmus milk | pink with whey 24 hrs |
Salt agar growth
| No growth |
Catalase Test
| bubbles after 5% H2O2 |
Hydrogen Sulfide Production (Klig.& Peptone Iron Tests)
| yellow throughout no black |
Tryptophane Test (Indole)
| bloody red after Kovac's |
Urea Test
| not done |
Gelatin Test
| liquid after taken from incubator, solid after 30 min at 5C |
ONPG test
| yellow |
Litmus Milk
| pink with whey |
Kirby Antibiotic Sensitivity Test: sensitive to...
| not done |
Enterotube II Test for Gm- oxidase - rods only
| not done |
Oxiferm Tube Test for Gm- oxidase + rods only
| not done |
Antibiotic Disc Test
| not done |
Antiseptic/Disinfectant Disc Test
| not done |
UV Light Sensitivity Test
| not done |
Tellurite Glycine Agar
| not done |
What is Unknown #B GENUS:________________ 10 pts ____________ species? ( 3 more Pts)
This assignment is due TYPED/handwritten table input is OK usually before the MIDTERM. The Bonus will be added to your course grade unless otherwise announced in class!
All students MUST complete this assignment but only those papers submitted before the deadline will receive the bonus! You are encouraged to work in groups and turn-in a GROUP PAPER with all your names (alphabetized by last name) and course Section # in the upper right corner of the cover page as well as the DATE submitted.
All students MUST complete this assignment but only those papers submitted before the deadline will receive the bonus! You are encouraged to work in groups and turn-in a GROUP PAPER with all your names (alphabetized by last name) and course Section # in the upper right corner of the cover page as well as the DATE submitted.
To turn-in this assignment: in the upper right-hand corner of the 1st page - (1) Type your LAST NAME, first name (as many as 4 people can work together and hand-in 1 paper - BUT YOU ALL MUST DISCUSS IT AND sign a statement that you discussed how to do it) (2) on the second line put your class designation and section number (3) on the 3rd line put the term submitted.
*NOTE: You must put a statement saying you participated in the discussion of this assignment and assisted with the preparation of the submitted paper in a "material" way. Put the statement at the bottom of the assignment cover page
The purpose of this exercise is to teach you how to FIND the reference material for your unknown paper in a library and to teach you how to INTERPRET the reference reading material in a way useful to the proper completion of your unknown paper...
NOTE: I will NOT tell you if you are correct or confirm your Genus during the exercise as there are many people who "communicate" too much. Just DO IT! THE PROCESS is more important to me than the answer...
Good Luck!
DRH
CLICK HERE TO LEARN: How to use a Dichotomous Key
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