Select and Inoculate Culture Media

Question 1

What helps you select the appropriate medium?

Answer 1

What was learned on the initial Gram staining

Question 2

What else can you use to select the appropriate medium?

Answer 2

Oxygen requirements, and Nutrition

Question 3

Oxygen requirements

Answer 3

Obligate aerobes
Obligate anaerobes
Facultative anaerobes
Microaerophilic
Capnophilic

Question 4

Obligate aerobes

Answer 4

Bacteria that require oxygen to survive

Question 5

Obligate anaerobes

Answer 5

Bacteria that are killed in the presence of oxygen or those with growth that is inhibited in the presence of oxygen

Question 6

Facultative anaerobes

Answer 6

Organisms that can survive in the absence of oxygen, but their growth is limited

Question 7

Microaerophilic

Answer 7

Bacteria that prefer reduced oxygen tension

Question 8

Capnophilic

Answer 8

Bacteria that require high levels of carbon dioxide

Question 9

Fastidious microbes

Answer 9

Picky eaters

Question 10

Agar

Answer 10

Dried sea algae extract

Question 11

General purpose media

Answer 11

Aka nutrient

Not common in vet med

Question 12

Enriched media

Answer 12

Meets requirements of fastidious pathogens

Question 13

What does enriched media meet the requirements of?

Answer 13

Fastidious pathogens

Question 14

Selective media

Answer 14

Contains antibacterial substances

Question 15

Differential media

Answer 15

Allows bacteria to differentiate into groups by biochemical reaction

Question 16

Enrichment media

Answer 16

Favors the growth of a particular type/group of organisms

Question 17

Transport media

Answer 17

Keeps microbes alive but does not encourage growth

Question 18

What two media is Blood Agar?

Answer 18

Enriched (fastidious) and Differential (hemolysis)

Question 19

What does Blood agar do?

Answer 19

An Enriched medium that is used to grow a wide variety of bacteria and a Differential medium based on hemolysis

Question 20

What are the three different types of hemolysis seen with blood agar

Answer 20

Alpha
Beta
Gamma
Delta

Question 21

Alpha hemolysis

Answer 21

Partial hemolysis

Question 22

Beta hemolysis

Answer 22

Complete hemolysis

Creates a clear zone around the colony

Question 23

Gamma hemolysis

Answer 23

Hemolysis that produces no change in the appearance of the medium and around the colonies

Question 24

Delta Hemolysis

Answer 24

Double zone hemolysis

Question 25

What two media is MacConkey Agar?

Answer 25

Selective and Differential

Question 26

What kind of bacteria does selective media grow?

Answer 26

Gram negative

Question 27

Lactose fermenting

Answer 27

Bacteria changes pink

Question 28

Non-lactose fermenting

Answer 28

Bacteria does not change color

Question 29

Eosin-Methylene Blue Agar (EMB)

Answer 29

A selective and differential medium used to isolate fecal coliforms

Question 30

What does Eosin-Methylene Blue allow to grow?

Answer 30

Gram positive bacteria Escheria coli Klebsiella pneumonia

Question 31

What color does E.coli and K. pneumonia colonies turn in a Eosin-Methylene Blue Agar?

Answer 31

Metallic Green

Question 32

Thioglycollate broth

Answer 32

An enrichment broth used to determine oxygen requirements of microorganisms

Question 33

Mueller-Hinton Agar

Answer 33

A general purpose medium used for culture and sensitivity testing of rapid growth bacteria

Question 34

What is the Goal of inoculation?

Answer 34

Isolated colonies

Question 35

How do you start the inoculation process?

Answer 35

Flame the wire loop and allow it to cool. | Collect a small amount of bacteria from an isolated colony from 1st agar plate, broth, or swab

Question 36

What happens after you collect the bacteria for inoculation?

Answer 36

Make streak area 1 then flame the loop from near to distal

Question 37

What do you with the loop once it is flamed?

Answer 37

Drag it once through area 1 and streak area 2.

Question 38

After streaking the loop through area 2 what is the next step in inoculating?

Answer 38

Flame the loop and drag once through area 2 and make area 3

Question 39

What do you do if you are doing the quadrant method?

Answer 39

Make area 4 by flaming the loop and label

Question 40

How do you place the agar in the incubator?

Answer 40

Inverted

Question 41

How do you remove the bacteria to inoculate the slant and butt?

Answer 41

With a small amount of bacteria using a sterile bacteriologic needle

Question 42

Where do you stab the needle when inoculating a slant and butt?

Answer 42

Directly into the center of the agar tube. - Push the needle all the way down to the bottom - Withdraw the needle through the same way

Question 43

How do you streak a slant and butt?

Answer 43

Using a back n forth motion starting at the bottom of the slant

Question 44

What temperature should agar plates be incubated at?

Answer 44

37 degrees celsius

Question 45

Why should agar plates be placed upside down?

Answer 45

To lessen the contamination risk from airborne particles landing on them Prevent water condensation

Question 46

Mesophiles

Answer 46

Grow in moderate temperature

Question 47

Psychrophiles

Answer 47

Grow at a lower temperature

Question 48

Thermophiles

Answer 48

Grow in a higher temperature

Question 49

How do bacteria reproduce?

Answer 49

Binary fission

Question 50

What are the four phases of bacterial growth?

Answer 50

1. Initial/Lag phase 2. Exponential 3. The stationary phase 4. The death/final phase

Question 51

Initial/Lag phase

Answer 51

Adjust to the environment

Question 52

Exponential

Answer 52

Increase in number of living bacterial cells, until space is used up or wastes accumulate

Question 53

The Stationary phase

Answer 53

Plateau in the number of living bacterial cells

Question 54

The Death/final phase

Answer 54

Exponential decrease of living cells

Question 55

What stage are spores produced at?

Answer 55

The death/final phase

Question 56

What are the characteristics of colonies?

Answer 56

``` Size Pigment Density Elevation Form Texture Odor Hemolysis ```

Question 57

How do aerobic bacteria produce energy

Answer 57

With the use of oxygen

Question 58

How do anaerobic bacteria produce energy?

Answer 58

Without oxygen

Question 59

How is culturing anaerobes different than aerobes?

Answer 59

Anaerobic cultures need to be collected more quickly than aerobic cultures

Question 60

What is else is different in culturing anaerobes and aerobes?

Answer 60

The growth medium

Question 61

What is the growth medium for anaerobes?

Answer 61

Thioglycollate

Question 62

What is the growth medium for aerobes?

Answer 62

Agar