Chapter 3.1

Question 1

The Five I’s

Answer 1

-Inoculation
-Incubation
-Isolation
-Inspection
-Identification

Question 2

Inoculation

Answer 2

Introduction of microbes into or upon a media

Question 3

Culture

Answer 3

-(verb) Adding to and the growing of microbes in various media
-(Noun) Population of microbes in or on nutrient medium

Question 4

Medium/Media (plural)

Answer 4

A nutrient-containing environment in which microbes can multiply

Question 5

Incubation

Answer 5

Inoculated media placed in temp controlled chamber called incubators

Question 6

Incubator temperatures and function

Answer 6

-Control temperature for culture growth.
-Temps ranging from 20-45 degrees C
-Also control atmospheric gasses ( oxygen, CO2)

Question 7

Microbial growth in liquid medium is

Answer 7

Cloudy or has turbidity

Question 8

Colonies

Answer 8

Visible masses of piled up cells

Question 9

Media classified into 3 properties

Answer 9

1) Physical state 2) Chemical Composition 3) Functional type

Question 10

Media Physical state

Answer 10

Liquid, semisolid, solid

Question 11

Media Chemical composition

Answer 11

Chemically defined (synthetic) or Complex (not chemically defined)

Question 12

Media Functional types (8)

Answer 12

-General purpose
-Enriched
-Selective
-Differential
-Anaerobic growth
-Specimen transport
-Assay
-Enumeration

Question 13

Liquid Media

Answer 13

-Water based solutions that do not solidify above freezing temps
-Broths, milks, infusions

Question 14

Semisolid media

Answer 14

-Clot like consistency at room temps
-Contains enough agar or gelatin to thicken but not solidify
-Used to determine motility of bacteria
-Also used to localize a reaction to specific site

Question 15

Brain and heart infusion

Answer 15

• made by boiling cow or pig heart and brain. Very nutrient dense
  semisolid- lower concentration of agar (0.5-1%)

Question 16

Solid Media

Answer 16

-Provides a firm surface upon which cells can form discrete/separate colonies
-Used to isolate bacteria and fungi

Question 17

Agar media

Answer 17

-Complex polysaccharide isolated from red alga
-Solid at room temp, liquefies at 100 C
-Begins to solidify at 42 C
-Any medium containing 1-5% agar usually has the word “agar” in its name

Question 18

Defined (synthetic) Media

Answer 18

-Exact chemical compositions are known
-May contain pure organic and inorganic compounds that vary little from one source to another

Question 19

Complex Media

Answer 19

-Contains at least one component that is not chemically definable.
-Provides rich mix of nutrients for microbes that have complex needs
examples: Blood, serum, meat extracts,milk, yeast extract, soybean

Question 20

Fastidious bacteria

Answer 20

Have complex or specific growth requirements

Question 21

General purpose media

Answer 21

-Complex media w/ nutrients that grow a broad spectrum of microbes

-Nutrient agar and broth, brain heart infusion, and soy agar

Question 22

Enriched media

Answer 22

-Contains complex organic substances that fastidious bacteria require for growth.
(blood, serum, hemoglobin or special growth factors)

Question 23

Selective media

Answer 23

-Contains one or more agents that inhibit growth of certain microbes.
-Encourages select microbes to grow
-Primary isolation of a certain type of microorganism from a mixed sample

Question 24

Differential Media

Answer 24

-Allow multiple types of organisms to grow but display visible differences between species
(size, color, media color changes or formation of gas bubbles or precipitates)

Question 25

Blood Agra

Answer 25

-Differential AND enriched media -used as an enrichment medium for fastidious microbes as well as differential for seeing visible differences

Question 26

Hemolysins

Answer 26

Enzymes that (break down) red blood cells to release iron-rich hemoglobin

Question 27

Beta- hemolysis

Answer 27

-Complete lysis of red blood cells -Transperency

Question 28

Alpha- hemolysis

Answer 28

incomplete lysis of red blood cells

Question 29

Gamma hemolysis

Answer 29

no hemolysis

Question 30

MacConkey Media

Answer 30

-Selective for gram negative bacteria -Lactose fermenters grow red whereas non-fermenters grow yellow

Question 31

Mannitol Salt Agar

Answer 31

-Selective for staphylocci - Mannitol fermenters turn medium yellow. -nonfermenter growth turns pink

Question 32

Reducing Media

Answer 32

Grow anaerobic bacteria

Question 33

Carbohydrate fermentation media

Answer 33

-Contain sugars that can be fermented and pH indicator to show reaction

Question 34

Transport media

Answer 34

Maintains and preserves specimen

Question 35

Assay Media

Answer 35

-Tests effectiveness of antimicrobial drugs disinfectants, antiseptics, cosmetics and preservatives on the growth of microorganisms

Question 36

Enumeration

Answer 36

-Used in industrial and environmental microbiology to count the numbers of organisms in milk, water, food, soil etc

Question 37

Isolation

Answer 37

Individual bacterial cell separated and given space on a firm nutrient surface.

Question 38

Colony

Answer 38

-Mound of cells formed on solid nutrient surface. -Arises from a single cell

Question 39

Isolation techniques

Answer 39

-Streak plate method -Pour Plate Method -Spread Plate Method

Question 40

Pure culture/Anexic

Answer 40

Container of medium that contains only a single known species or type of microorganism

Question 41

Subculture

Answer 41

A second level culture from a well isolated colony

Question 42

Mixed culture

Answer 42

Container that holds two or more identified, easily differentiated species of microbes.

Question 43

Contaminated culture

Answer 43

A culture that was once pure or mixed that now contains contaminants (unwanted microbes)

Question 44

Inspection

Answer 44

-Examining growth and location of growth in tubes -Looking at morphology of cells or colonies (size, shape, color)

Question 45

Identification

Answer 45

Identifying celluar geneses, species and metabolism through -Biochemical testing -Genotypic testing (DNA) -Immunological testing

Question 46

Biochemical testing

Answer 46

Determines chemical characteristics such as nutrient requirements, products given off during growth, presence of enzymes, and mechanisms for deriving energy.

Question 47

Genotypic testing

Answer 47

Analysis of DNA-PCR sequencing

Question 48

Immunologic testing

Answer 48

Testing against known antibodies