Microbiology Quiz Basic Lab Tests © NursingJourney.com Microbiology Quiz Basic Lab Tests FYI, in this quiz there may be words used such as slant, but the photo doesn’t show the media as a slant. Therefore, just think about what you use in your lab, and relate that to what you are seeing here. Photos have been “borrowed” from the internet. If they are yours, please e-mail your URL so that I can give you credit. Fun2Care@yahoo.com Also, most information was taken from Alfred E. Benson’s Microbiology Applications, Short Version, 9th Edition

In a Gram stain, what are the positive and negative results? © NursingJourney.com In a Gram stain, what are the positive and negative results?

In a Gram stain, what are the positive and negative results? © NursingJourney.com In a Gram stain, what are the positive and negative results? Positive: Purple Negative: Pink

In a spore stain, what are the positive and negative results? © NursingJourney.com In a spore stain, what are the positive and negative results?

In a spore stain,what are the positive and negative results? © NursingJourney.com In a spore stain,what are the positive and negative results? Positive: Green spores Negative: No spores (pink)

In a acid-fast stain,what are the positive and negative results? © NursingJourney.com In a acid-fast stain,what are the positive and negative results?

In a acid-fast stain,what are the positive and negative results? © NursingJourney.com In a acid-fast stain,what are the positive and negative results? Positive: Pink Negative: Blue

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? http://faculty.southwest.tn.edu/bdrake/Microbiology%20Pictures.htm

Motility Test Media - MTM © NursingJourney.com What is this? What do the results reveal? Motility Test Media - MTM Negative for Motility Growth is confined to the stab area. Positive for Motility Growth is radiating from the stab area, cloudiness is present http://faculty.southwest.tn.edu/bdrake/Microbiology%20Pictures.htm To test if the organism is motile, stab a depth of 75% of the media and incubate 24-48 hours at the best growth temperature already decided.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal?

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Gelatin Liquefaction - Nutrient Gelatin Negative result:-Solid in ice bath or refrigerator after 5 minutes. Positive result: Liquid in ice bath or refrigerator after 5 minutes. Stab straight to bottom with an inoculating needle. Put in ice bath or refrigerator for 5 minutes, read results. If negative, reincubate up to 30 days.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal?

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Phenylalanine Deaminase - Phenylalanine agar slant (PA) Positive result Green slant in 0-10 minutes Streak slant with inoculator, After incubation time, use 5 drops of phenalalanine reagent. Negative result Yellow slant after 5-10 minutes

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal?

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Glucose, Lactose or Mannitol Fermentation - Phenol Red, Glucose, Lactose Mannitol Broth Negative Result: Red-pink (negative for acid) No bubbles (negative for gas) Positive Result: Yellow (positive for acid) Bubbles (positive for gas) Use Durham tube to test for gas, incubate 24-48 hrs. Reincubate negative results up to 30 days.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal?

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Methyl Red Test Positive Result Negative Result After incubating for 2-5 days, add 3-4 drops of Methyl Red reagent.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal?

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Urea hydrolysis - Urea broth Positive Result Negative Result After heavy inoculation, incubate up to 30 days.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal?

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Starch hydrolysis - Starch agar plate (SA) Positive Result Clear zone Negative Result Dark zone After incubating 24-48 hrs, add grams iodine over growth.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal?

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Citrate utilization - Simmon’s citrate agar slant (S/C) Negative Result Positive Result Uses citrate as sole carbon source Streak slant and stab butt, Incubate up to 30 days.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal?

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Tryptophan hydrolysis (indole) - Tryptone broth Positive Result Red ring Negative Result Yellow ring 10 drops of Kovac’s reagent are added after a 24-48 hour incubation.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal?

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Casein hydrolysis - Skim milk agar plate (SMA) Positive Result Clear zone Negative Result no zone Incubate for 24-48 hours

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? http://smccd.net/accounts/case/biol240/vlab.html

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Litmus reactions - Litmus milk A = Alkaline C = Acid D = Litmus Reduction E = Peptonization (Another result may be coagulation, but is hard too distinguish in a photo.) http://smccd.net/accounts/case/biol240/vlab.html 1-5 days incubation time

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? http://biology.uwsp.edu/faculty/TBarta/

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Hydrogen Sulfide Production - Kliger’s iron agar slant (KIA) Lactose Negative Red Slant Glucose Positive Yellow butt Hydrogen Sulfide Positive Black butt http://biology.uwsp.edu/faculty/TBarta/ Streak slant for lactose, stab butt for glucose. Incubate 24-48 hours. Save in refrigerator for new working stock. Lactose positive would be a yellow slant, and Glucose negative would be a red butt. If it is H2S+ negative, there will not be any black visible.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? http://people.uleth.ca/~selibl/Biol3200/BiochTests/

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Voges-Proskauer Test, aka Acetoin - 1/2 MR-VP media Voges-Proskauer Negative Brown color Voges-Proskauer Positive Red color http://people.uleth.ca/~selibl/Biol3200/BiochTests/ Add a dropperful of Barritt’s reagent A&B, after incubating for 5 days, and shake well for 2 hours.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? http://biology.uwsp.edu/faculty/TBarta/

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Oxidase Production - Nutrient agar plate (NA) Oxidase Negative No black colonies Oxidase Positive Black colonies within 30 minutes http://biology.uwsp.edu/faculty/TBarta/ After incubation, tamp one oxidase disk on first quadrant

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? http://faculty.southwest.tn.edu/bdrake/Microbiology%20Pictures.htm

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Catalase Production - Nutrient agar slant (NA) Catalase Production Negative No bubbles Catalase Production Positive Bubbles http://faculty.southwest.tn.edu/bdrake/Microbiology%20Pictures.htm After a working stock has been incubated for 3-5 days, add 3 drops of hydrogen peroxide. Make sure to use an OLD working stock for this test!

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? http://academic.mwsc.edu/jcbaker/images/nitrate%20reduction.jpg

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Nitrate Reduction - Nitrate agar slant Postive for Niatrate Reduction Red color Negative for Nitrate Reduction Clear color http://academic.mwsc.edu/jcbaker/images/nitrate%20reduction.jpg Incubate, and then add 3 drops of Nitrate reagents A & B. If a negative result occurs, confirm the result with zinc dust. Opposite color results will occur: Clear positive, red negative for nitrate reduction.

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? http://academic.pg.cc.md.us/~kroberts/lecture/web/exoenzymes/lipase.jpg

What is this? What do the results reveal? © NursingJourney.com What is this? What do the results reveal? Fat Hydrolysis - Spirit Blue agar plate (SBA) Postive for Fat Hydrolysis Blue precipitate Negative for Fat Hyrolysis No precipitate http://academic.pg.cc.md.us/~kroberts/lecture/web/exoenzymes/lipase.jpg Incubate for 24-48 hours.

Calculate the following dilution: © NursingJourney.com Calculate the following dilution:

Calculate the following dilution: © NursingJourney.com Calculate the following dilution: The correct answer is: C. (Remember to round the numbers.)

Calculate the following dilution: © NursingJourney.com Calculate the following dilution: http://smccd.net/accounts/case/biol230/dilutions2.html#

Calculate the following dilution: © NursingJourney.com Calculate the following dilution: 10-2 10-4 10-5 10-6 http://smccd.net/accounts/case/biol230/dilutions2.html# The answer is 10-6, or .000001

Calculate the following dilution: © NursingJourney.com Calculate the following dilution: http://smccd.net/accounts/case/biol230/dilutions2.html#

Calculate the following dilution: © NursingJourney.com Calculate the following dilution: 10-1 10-2 10-3 http://smccd.net/accounts/case/biol230/dilutions2.html# The answer is 10-3, or .001

Microbiology Quiz Basic Lab Tests © NursingJourney.com Microbiology Quiz Basic Lab Tests Congratulations!! That’s all for this Microbiology quiz! Please remember, your instructor may require you to learn more, or less, of what is on this PowerPoint. It’s your responsibility to learn what you will be tested on!! (Including knowing if any errors were made.) Please report any errors to Fun2Care@Yahoo.com.