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Micro 225 Lab Exam #3 final

front 1

Lipid Hydrolysis Test is used in biochemical test methods for bioluminescent marine vibrio & Photobacterium species, what medium is used and what are we testing for?

back 1

Biochemical test: Lipase
Medium used: Marian fat agar plate, tributyrin is added to agar
Red color of fat agar due to neutral red pH indicator

front 2

What would be the results for this Lipid Hydrolysis test?

back 2

Positive: colony turns red (may also turn red-pink around & below colony)
-Lipase produced
-free Fatty acids are liberated from hydrolysis of triglyceride
-Neutral red precipitates turn red from acidic pH of fatty acids
Aeromonas hydrophila = +

front 3

What would be the results for this Lipid Hydrolysis test?

back 3

Negative: no evidence of red color on or around colony
-No lipase production
-Neutral red remains as amber color from neutral pH (no color change)
Photobacterium phosphoreum= -

front 4

Starch Hydrolysis Test is used in biochemical test methods for bioluminescent marine vibrio & Photobacterium species, what medium is used and what are we testing for?

back 4

Biochemical test: presence of Amylase
Medium used: Marine starch agar plate
Flood area of spot inoculation with Gram's Iodine: Chemically reacts with starch =blue-black color

front 5

What would be the results for this Starch Hydrolysis Test?

back 5

Positive: clearing around colony (halo)
-Amylase produced
-Some starch digested
-NO binding of iodine around colony due to starch breakdown
Aeromonas hydrophila = +

front 6

What would be the results for this Starch Hydrolysis Test?

back 6

Negative= NO clearing around colony (no halo)
-NO amylase production
-No starch digested
-Iodine binds to starch on all parts of the plate
Photobacterium phosphoreum= -

front 7

Gelatin Hydrolysis test is used in biochemical test methods for bioluminescent marine vibrio & Photobacterium species, what medium is used and what are we testing for?

back 7

Biochemical test: Gelatinase produced
Gelatinase is an enzyme that liquefies gelatin
Medium used:Marin nutrient gelatin agar plate

front 8

What would be the results for this Gelatin Hydrolysis test?

back 8

Positive: liquefaction of gelatin
-Part or all does not re-solidify
-Gelatinase produced
-Gelatin digested
Aeromonas hydrophila = +

front 9

What would be the results for this Gelatin Hydrolysis test?

back 9

Negative: Gelatin remains solid
-Re-solidifies with refrigerated
-NO gelatinase produced
-NO Gelatin digested

Photobacterium phosphoreum= -
If NEGATIVE, continue incubation at unknown's preference temperature:
-Repeat refrigeration test each lab for up to 2 weeks
-Do not leave gelatin tubes in refrigerator

front 10

What biochemical test methods are used for utilization of Amino acids ?

back 10

Metabolism of lysine/ornithine by decarboxylation or arginine by dihydroxylation, then decarboxylation
-Lysine (LDC)
-Ornithine (ODC)
-Arginine (ADH)

front 11

What occurs during decarboxylation?

back 11

Removes acid groups- carboxyl groups (COOH) from amino acids & other organic molecules. Only activated by acidic environments: fermentation
-Neutralizes acids: survival mechanism; Amines released increases pH

front 12

What are the reactions that Lysine (LDC), Ornithine (ODC), and Arginine (ADH) undergo during utilization of amino acids?

back 12

Metabolism of lysine/ornithine by decarboxylation or arginine by dihydroxylation, then decarboxylation

front 13

What would be the Decarboxylation results for this utilization of Amino acids test ?

back 13

Positive Results:
DC=Yellow
-Glucose fermentation
-Acid produced
-Brom-cresol purple (pH indicator) turns yellow

LDC, ODC, ADH= Purple
-Acids from glucose fermentation stimulated decarboxylase
-Amine neutralizes acids
-pH indicator turns purple

front 14

What would be the Decarboxylation results for this utilization of Amino acids test ?

back 14

Negative results:
DC=Yellow
-Glucose fermentation
-Acid produced

LDC, ODC, ADH=Yellow
-Acids produced from glucose fermentation
-No Decarboxylase release
-No acid neutralization

front 15

What would be the Decarboxylation results for this utilization of Amino acids test ?

back 15

Negative results:
DC=purple
-Glucose not fermented
-Acids not produced

LDC, ODC, ADH= Purple
Decarboxylase no produced

front 16

What would be the results for this Voges proskauer Biochemical test?

back 16

Medium used: Methyl red voges proskauer broth (MRVP) + Marine cations 1559.
Preform test by adding KOH & Alpha mix ever few mins for entire lab
-measure the ability of an organism to produce non-acid products (neutral pH) from glucose

Results:
Voges Proskauer Positive= Butanediol Fermenter
If Reddish pink color slowly develops: Positive
P. phosphoreum= +

NO color change: Negative= Non-fermenter or fermentation pattern
Vibrio fischeri= -

-May take min-hours to develop leave with instructor in rack up front if still negative at the end of lab

front 17

What would be the results for this Idole Production Biochemical test?

back 17

Medium used: Peptone broth for idone +Marine cation 1559, contains tryptophan and can detect motility of organism
Perform test by Add Kovac's reagent to top

Positive= pink-red color on top
presence of Indole
-tryptophanase produced from tryptophan utilization
-Idole detected

Negative: No color change
Photobacterium phosphoreum= -

front 18

What is the function of Poly-B-hydroxybutyrate (PHB) granules?

back 18

Medium used: Marine Glucose Fermentation broth, view wet mount under phase contrast microscopy

-energy reserve-> Carbon and energy source
-Produced under high Carbon conditions for later

P. phosphoreum=-
V. logei & V. fischeri= +

front 19

Study of fungi:

back 19

Mycology

front 20

What are fungi?

back 20

Molds, yeasts, mushroom, relatives

front 21

Characteristics of fungi?

back 21

-Eukaryotes
-Nonphotosynthetic-heterotrophs
-Absorptive metabolism- Exoenzymes

front 22

Cell structure of Fungi?

back 22

-Lack tissue differentiation
-Cell wall of chitin/ other polysaccharides

front 23

Reproduction of Fungi?

back 23

Propagate by spores (sexual/Asexual)
-Deuteromycota division (defunct, term of convenience for non-sexually reproducing fungi) -most now in Ascomycota

front 24

Habitats of fungi?

back 24

Primary terrestrial, a few freshwater and marine

front 25

Ecology of fungi?

back 25

-Symbionts of plants or animals- commensals, mutualistic, parasitic
-Mutualist:
*Form associations with plant roots (mycorrhizae) - Assist with N & P uptake
* Association with Algae or cyanobacteria (lichens)

front 26

How do plants/trees benefit from presence of Mycorrhizae?

back 26

Mutualist:
*Form associations with plant roots (mycorrhizae) - Assist with N & P uptake
-Redwood seedling without (left) & with (right) mycorrhizae
-Pine seedling- mycorrhizal roots from one tree spread to inoculate other tree roots

front 27

Why is fungi important for the environment ?

back 27

Decomposers (saprophytes)- break down organic material & return to environment
-Pathogens: mycose of animals & plants

front 28

Why is fungi important industrially ?

back 28

Industrial fermentation- bread, wine, beer, cheese, tofu, soy sauce, steroid manufacture, antibiotic production

front 29

What is the cell structure of Molds?

back 29

multicellular filamentous fungi

front 30

What are the four major divisions of molds?

back 30

1. Zygomycota
2. Ascomycota
-Deuteromycota division (defunct, term of convenience for non-sexually reproducing fungi) -most now in Ascomycota
3. Basidiomycota
4. Chytridiomycota

front 31

What is the cell structure of Yeast?

back 31

-Thallus: fungal body or vegetative structure
-Chitin: N-containing polysaccharide (cell wall of some fungi)

front 32

What are the four major division of Yeast?

back 32

Ascomycota- also contains some molds

front 33

fill in the blanks

back 33

1. Plasma membrane
2. Chitin microfibrils
3. Cytoplasmic microtubule
4. Cluster of ribosomes
5. Smooth endoplasmic reticulum
6. Mitochondria
7. Rough ER
8. Cell wall
9. Meshwork of microfilaments
10. Plasma membrane invagainations

front 34

What is the difference between these hyphae cross walls?

back 34

Vegetative (reproductive) hyphae- structures that elongate into branching multicellular filaments
-Hyphae may be septate (contain dividing cross walls) or coenocytic (cross walls absent)

front 35

Label mold hyphae structure.

back 35

1. Cell wall
2. Pore
3. Nuclei
4. Septum
a) Septum hyphae
b) Coenocytic hyphae

front 36

What is aerial mycelium and how is it observed?

back 36

Hyphae intertwine to form a aerial mycelium on substrate surface- puffy colonies
-Vegetative mycelium seen on underside of plates

front 37

Label the diagram

back 37

1. Spore: non-motile reproductive units on end of hyphae- useful for ID (development & morphology)
c) Growth of a hypha from a spore

front 38

What specialized hyphae & spores is this (label structures)?

back 38

Conidiophores bear long chains of asexual conidiophores.
-conidiophores break away & can germinate to form new vegetative hyphae
1. Conidiophores
2. Conidiophore

front 39

What specialized hyphae & spores is this (label structures)?

back 39

Sporangiophores bear sac (sporangium) that contains sporangiospores.
1. Sporangiospores
2. Sporangium
3. Sporangiophores

front 40

What specialized hyphae & spores is this (label structures)?

back 40

Vegetative mother cells bear single blastospores that bud off.
1. Blastospores
2. Vegetative mother cell

front 41

What specialized hyphae & spores is this (label structures)?1

back 41

Arthrospores-hyphae fragment apart & the component function as spores.
a. Arthroconidia (arthrospores)
b. Fragment hypha

front 42

What specialized hyphae & spores is this (label structures)?

back 42

Transverse tissue forming new cell wall

front 43

What specialized hyphae & spores is this (label structures)?

back 43

a. Terminal chlamydospores
b. Chlamydospore within a hypha

front 44

Identify this cell shape:

back 44

Yeast structure; spherical, ellipsoidal, or oval
-Most much larger than bacteria: 3-4 um
-Except for dimorphic yeast (Candida form pseudohyphae) hyphae are not formed

front 45

identify these colonies:

back 45

Yeast colonies- appear like bacteria, often dull and opaque

front 46

Identify and label the structure

back 46

Yeast Cell:
1.Nucleus
2. Cytoplasm
3. Polar bud scar
4. Mitochondrion
5. Plasma membrane
6. Golgi apparatus
7. Nuclear envelope
8. Plasma membrane
9. Chromosome
10. Polar bud Scar
11. Cell wall

front 47

What term is used to describe some fungi alternate between a mold & yeast form: Hyphal cells --> yeast cells?

back 47

Dimorphism

front 48

label the structure:

back 48

a. Spores
b. Hyphae
c. spores
d. Bud
e. Yeast

1. Increased Temperature, Reduced O2, Suboptimal nutrients
1a. When fungal spores from the environment gain entrance to a warm-blooded animal , they germinate into yeast and remain in this phase in the host.
Animal Habitat: Parasitic, Yeast phase
Temp: 35-45 C)
Reproduction through: endospores

2.Decreased temperature, Optimal O2 content, Improved nutrients
2a. Yeast cells leaving the animal host return to the environment and revert to the sporulating hyphal stat. These conversions can be demonstrated on artificial media in the lab.
Natural Habitat:Saprobic (free-living), Mycelial phase
Tem: (<30 C)
Reproduction through: sporulation

front 49

In animal-infecting fungi:

back 49

alternate between a saprophytic mold phase in environment & parasitic yeast phase in host:
-Striking adaptation to permit growth & survival in mammalian host
-Thermal dimorphism-grow as mold at <30C as yeast at 35-40 C
*Candida albicans (thrush), Coccidioides (valley fever)

front 50

In plan-infecting fungi:

back 50

mold form in plant host, yeast form in environment

front 51

Identify and label the structure

back 51

The Morphology of Coccidioides
a. Free Arthrospores
b. Septate Mycelium
c. Arthrospore formation
d. Disarticulation
e. Immature Spherules
f. Endosporulation spherule (mature)
g. Free endospores
h.Rupture spherule

front 52

dentify and label the structures

back 52

A. Coccidioides immitis- filamentous colonie isolated from patient's sputum
B. immature and mature spherules in liver tissue from mice inoculated with the isolated filamentous fungus.
C. Histologic section with Gomori-Grocott silver impregnation

front 53

Identify and label the structure

back 53

a. Gamete release 1N
b. Fertilization 1N+1N
c. Zygote 2N
d. Young sporophyte 2N
e. Zoosporangium (Mitosporangium) 2N
f. Resistant sporangium (Meiosporangium) 2N
g. Meiospores 1N
h. Sporophyte 2N
i. Male gametangium
j. Female gametangium
k. Male gametes
l. Gametophytes 1N
m. Female gametes

front 54

Reproduction of mold:

back 54

-Many have sexual & asexual phases
-Sexual phase: hyphal structures- Gametoangia intertwine or fuse, forming sexual spores
-Asexual phase: form spores or conidia (reproductive units)

front 55

Reproduction of Yeast:

back 55

-Asexula reproduction by fission (splitting) budding
-Budding: daughter cell formed by parent cell-bud formed by small outpouching of cell
*Bud may separate from original cell or remain attached. Chain of buds may form if they remain attached. forming pseudohyphae (pseudomycelia). Each cell can form up 24 buds.
-SEXUAL reproduction in some yeast. Produce several types of spores (ascospores). Type of spore produced useful for yeast ID.

front 56

Commensal yeasts in Humans:

back 56

-Most yeast nonpathogenic, but opportunistic if host becomes immunosuppressed.
ex: Candida, Malassezia,

front 57

Candida:

back 57

Commensal Yeast of Humans:
normal flora or skin & mucous membranes (mouth)

front 58

Malassezia:

back 58

Dandruff & seborrhea (large flakes & itchy red area around scalp)- Caused by Pityrosporum (malassezia)

front 59

How is dandruff prevented?

back 59

Zinc pyrithione, ketoconazole, Selenium- sulfide in dandruff shampoos inhibit yeast growth & some help inhibit cell turn over.

front 60

How can Fungi be cultivated?

back 60

on plates; molds & yeast colonies cultivated in manner similar to bacteria, but often slower to grow.

front 61

Why is Sabouraud's dextrose agar (SAB) 4% sugar and pH=5.6 used to grow fungi?

back 61

Because fungi and bacteria compete, selective media favoring fungi is used inhibits many bacterial due to osmophilic & acidophilic environment

front 62

Why are Henrici slides used for to analyze fungi?

back 62

mold can be studied during growth, without disturbing vegetative & reproductive structures.

front 63

Fungi are members of the domain
A) Archaea
B) Bacteria
C) Eukarya
D) Classificationisundetermined

back 63

C) Eukarya

front 64

Fungi reproduce
A) A sexually only
B) Sexuallyonly
C) Bothsexuallyandasexually
D) Type of reproduction is undetermined

back 64

C) Bothsexuallyandasexually

front 65

Which of the following nutritional types best describes the fungi?
A) Heterotrophic
B) Phototrophic
C) Saprophytic
D) Noneoftheabove

back 65

C) Saprophytic

front 66

Fungi are important ecologically because of their ability to
A) Fixatmosphericnitrogenandmakeitavailabletootherorganisms
B) Decompose organic matter to release carbon and nutrients to other organisms
C) Cyclesulfurandiron
D) A ll of the above

back 66

B) Decompose organic matter to release carbon and nutrients to other organisms

front 67

Symbiotic relationships between vascular plant roots and fungi are called
A) Hyphae
B) Mycelium
C) Zooxanthelle
D) Mycorrhizae

back 67

D) Mycorrhizae

front 68

Fungi are producers of which of the following industrial products?
A) Breadsandcheeses
B) Organicacids
C) A ntibiotics and immunosuppressive drugs
D) A ll of the above

back 68

D) A ll of the above

front 69

The simplest group of fungi are the
A) Chytridiomycota
B) Zygomycota
C) Glomeromycota
D) Ascomycota

back 69

A) Chytridiomycota

front 70

When food becomes scarce or environmental conditions become unfavorable, Rhizopus bread mold
A) Reproducessexually
B) Reproducesasexually
C) Doesnotreproduce
D) Noneoftheabove

back 70

A) Reproducessexually

front 71

Which of the following groups of fungi degrade many chemically stable organic compounds including lignin, cellulose, and collagen?
A) Chytridiomycota
B) Zygomycota
C) Glomeromycota
D) Ascomycota

back 71

D) Ascomycota

front 72

A unicellular fungi that reproduces by budding or binary fission is known as a:
A) Mold
B) Ascus
C) Yeast
D) Hyphae

back 72

C) Yeast

front 73

When nutrients are limited, diploid yeast cells undergo
A) Senescence
B) Sporulation
C) Mitosis
D) Meiosis

back 73

D) Meiosis

front 74

Filamentous ascomycetes reproduce by producing
A) Conidia
B) Sclerotia
C) Basidia
D) Asci

back 74

A) Conidia

front 75

Aspergillus mold, the trigger of allergies, asthma, and sinusitis, belongs to the fungal group
A) Chytridiomycota
B) Zygomycota
C) Glomeromycota
D) Ascomycota

back 75

D) Ascomycota

front 76

Ergotism, the toxic condition in humans and animals that eat grain infected with fungus, is caused by an organism from the group
A) Chytridiomycota
B) Zygomycota
C) Glomeromycota
D) Ascomycota

back 76

D) Ascomycota

front 77

Which fungal group includes the stinkhorns, puffballs, toadstools, and mushrooms?
A) Basidiomycota
B) Zygomycota
C) Glomeromycota
D) Ascomycota

back 77

A) Basidiomycota

front 78

Mushroom toxins damage the host by
A) A ttacking liver cells
B) Damagingcellsliningthestomachandintestines
C) Inhibiting RNA polymerase
D) A ll of the above

back 78

D) A ll of the above

front 79

Important plant pathogens causing "rusts" and "smuts" in crop plants are members of which fungal group?
A) Chytridiomycota
B) Basidiomycota
C) Glomeromycota
D) Ascomycota

back 79

B) Basidiomycota

front 80

Fungi can be unicellular or multicellular.
A) True
B) False

back 80

A) True

front 81

Fungi are saphrophytes and use hydrolytic enzymes to degrade organic material.
A) True
B) False

back 81

A) True

front 82

Flagella are absent in all fungi.
A) True
B) False

back 82

B) False

front 83

A fungus in the genus Rhizopus is the causative agent of seedling blight, a rice disease.
A) True
B) False

back 83

A) True

front 84

Mushrooms used in food production belong to the Basidiomycota.
A) True
B) False

back 84

A) True

front 85

Infection by some members of the Basidiomycota triggers tumor formation in crop plants.
A) True
B) False

back 85

A) True