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BMED-4440-CB Final

front 1

The Enterotube II / EnteroPluri-Test is designed to identify ___ bacteria and other select oxidase-______, Gram-_____ bacteria.

back 1

1. Enteric

2. Negative

3. Negative

front 2

How many chambers are there? Name the mediums present in each one.

back 2

12 chambers

  1. Glucose
  2. Lysine
  3. Ornithine
  4. H2S / Indole
  5. Adonitol
  6. Lactose
  7. Arabinose
  8. Sorbitol
  9. VP
  10. Dulcitol / PA
  11. Urea
  12. Citrate

front 3

E. coli Positives: Lys, Ind, Adon, Arab, Sorb

E. aerogenes Positives: Lys, Orn, Arab, Sorb, VP, Citrate

K. pneumoniae Positives: Lys, Arab, Sorb, VP, Urea, Citrate

What are the five digit codes?

back 3

E. coli: 04660

E. aerogenes: 06061

K. pneumoniae: 04063

front 4

For the Indole Test, what reagent is used? Positive result is what color?

back 4

Kovac's Reagent

Red

front 5

For the VP test, what reagents are used? Positive result is what color?

back 5

KOH Reagent & alpha-Naphthol Reagent

Red

front 6

Steps for performing the Enterotube Test?

back 6

  1. Pick up a colony
  2. Pull wire through
  3. Reinsert wire
  4. Break wire
  5. Puncture air inlets
  6. Add reagents AFTER incubation

front 7

ELISA is an acronym for

back 7

Enzyme Linked Immunosorbent Assay

front 8

It is used to ______

back 8

detect antigen or antibody in a sample

front 9

What are the steps to the ELISA test?

back 9

  1. The purified antigen is added to the wells of the strip, wait 5 min. to allow it to absorb to the plastic wall of the wells.
  2. Wash
  3. Serum is added, wait 5 min. to allow antibodies to bind to antigen coating.
  4. Wash
  5. Secondary antibody is added, wait 5 min. to allow it to bind to the serum's antibodies.
  6. Wash
  7. Add enzyme
  8. Observe result

front 10

Why is the secondary antibody needed?

back 10

It will be what gives us a positive or negative result.

front 11

What is the enzyme used and why is itneeded? What color is a positive result?

back 11

The enzyme is HRP and it will bind secondary antibody to indicate its presence. Blue indicates a positive result.

front 12

Where does the Kirby-Bauer method get its name?

back 12

Of the names who published it

front 13

What is the meaning of MIC?

back 13

Minimum inhibitory concentration

front 14

How is the antibiotic used in the Kirby-Bauer method?

back 14

Antibiotic-impregnated paper disks

front 15

What is the drug that kills bacteria?

back 15

Bactericidal

front 16

What is the drug that stops the bacteria from dividing?

back 16

Bacteriostatic

front 17

What do you read with McFarland turbitidy?

back 17

Cell Density

front 18

We diluted the cultures with sterile saline until ____ McFarland.

back 18

0.5

front 19

If a bacteria is susceptible to an antibiotic, the zone diameter would be ___.

What if it was resistant?

back 19

Large

Small

front 20

What bacteria did we use for the Kirby-Bauer lab?

back 20

E. coli and S. aureus

front 21

What antibiotics did we use for the Kirby-Bauer lab?

back 21

Chloramphenicol, Ciprofloxicin, Trimethoprim, Penicillin

front 22

What is bacterial transformation?

back 22

The process by which competent bacterial cells pick up DNA from the environment and make use of the genes it carries.

front 23

What are the structural and functional genetic units of prokaryotes?

back 23

Operons

front 24

Each operon minimally includes a ______ ______ and two or more ______ _______ coding for enzymes in the same metabolic pathway.

back 24

promoter site

structural genes

front 25

What is used as a vector in the pGLO lab?

back 25

a plasmid

front 26

Plasmids are _______

back 26

small, naturally occurring, circular DNA molecules that possess only a few genes.

front 27

What does the pGLO plasmid contain?

back 27

araC - allows RNA polymerase to bind to promoter

Pbad - arabinose promoter

GFP - green fluorescent protein, makes it glow

bla - antibiotic resistance gene, produces beta-lactamase which hydrolyzes ampicillin

ori - replication origin

front 28

Viruses that attack bacteria are called

back 28

bacteriophages

front 29

Once assembly is complete, the cell lyses and releases the phages, which then attack other bacterial cells. What is this process called?

back 29

Lytic Cycle

front 30

Lysis of bacterial cells growing in a lawn on an agar plate produce a clearing, what are these clearings called?

back 30

Plaques

front 31

Why are plaques important?

back 31

They allow us to calculate the phage concentration in a given sample.

front 32

What is the formula to calculate phage titer?

back 32

Phage titer = PFU / Original Sample Volume

front 33

Why is the soft agar, the tube that contains phage-host mix, important?

back 33

The consistency of it is sufficient enough to immobilize the bacteria while allowing the smaller bacteriophages to diffuse short distances and infect surrounding cells.

front 34

Which parasite causes an infection that involves a significant amount of surface area of the small intestine and can cause chronic diarrhea, dehydration, abdominal pain and other symptoms?

back 34

Giardia lamblia

front 35

What parasite causes malaria?

back 35

Plasmodium spp.

front 36

Review bacteria shapes & stains:

What does gram positive cocci look like?

What does gram positive bacilli look like?

What does gram negative spirochetes look like?

Be aware of difference between spirochetes, spirilla, and vibrio.

back 36

Purple circles

Purple rods / pill shaped

Pink coils

look up differences on google

front 37

Steps for Gram Staining

back 37

  1. Fixation
  2. Crystal Violet, 1 min
  3. Iodine, 1 min
  4. Decolorization
  5. Counter Stain (Safranin), 1 min

front 38

Steps for a Wet Mount

back 38

1. Place a loopful of water on a clean glass slide.
2 Gently add bacteria to the drop by placing the loop over the drop for several seconds.
3 Gently lower a cover glass with your loop supporting one side over the drop of water. Avoid trapping air bubbles.
4 Observe under high-dry power

front 39

What are the chemicals that develop color as they oxidize?

back 39

Chromogenic reducing agents

front 40

What kind of culture do you have if you only have a single species?

back 40

pure culture

front 41

What kind of culture do you have if you have two or more species?

back 41

mixed culture

front 42

What is an important thing to do with the metal material when transferring microorganisms?

back 42

sterilize using bunsen burner

front 43

When do you use petroleum jelly?

back 43

Hanging Drop

front 44

What color should a positive and negative result for a McConkey agar look like?

back 44

Pink for Positive, Yellow for Negative

front 45

What enzyme transforms hydrogen peroxide into water and gaseous oxygen?

back 45

Catalase

front 46

In the Oxidase Reaction, what enzyme turns the color?

back 46

cytochrome c oxidase

front 47

What substance is designed to reduce the number of pathogens in living tissue?

back 47

Antiseptics