Biology Laboratory Final Exam

Front 1

organisms that are the most widely distributed, simplest in morphology, smallest in size, most difficult to classify, hardest to identify

Back 1

Bacteria

Front 2

6 characteristics of bacteria

Back 2

1) Ubiquitous
2) Simple morphology (shape)
3) Small size (microscopic)
4) difficult to classify and identify
5) prokaryotic, belongs to kindgom Monera
6) Generally non-photosynthetic

Front 3

Proper labeling of a plate

Back 3

1) TA name
2) What is incubated?
3) Date
4) Media name
5) Initials

Front 4

any object, surface, or even a body part

Back 4

Fomite

Front 5

Most organisms are _____________ and do not reside on a particular host

Back 5

Free living

Front 6

any area where a microbe resides and serves as a potential source of infection

Back 6

Reservoir

Front 7

Most microorganisms are __________________; they are normally harmless but can cause infection when given the chance

Back 7

Opportunistic pathogens

Front 8

- unpigmented, motile organisms
- belongs to Kingdom Protista
- eukaryotic
- single-cell transparent organisms, cell membrane, no cell wall
- in lab, paramecium are most common to be seen

Back 8

Protozoa

Front 9

- greenish or golden brown
- belongs to Kingtom Protista and includes all photosynthetic eukaryotic organisms
- no tissue differentiation
- distinct, visible nuclei and chloroplasts
- unicellular or colonial

Back 9

Algae

Front 10

- blue-green
- belongs to Kingdom Monera
- prokaryotic type of nucleus
- phototrophic bacteria: bacteriochlorophyll

Back 10

Cyanobacteria

Front 11

allows one to see cellular structure of bacteria

Back 11

staining

Front 12

Smears should do this

Back 12

- withstand washings
- not be too thick
- not result in excessive distortion of organism

Front 13

aggregate of millions of cells that come from one cell

Back 13

Colony

Front 14

Purpose of heat-fixing

Back 14

- to kill bacteria
- to fix them to the slide
- to coagulate cytoplasmic proteins to make cells more visible

Front 15

using a single stain to color bacteria due to their transparent nature

Back 15

Simple staining

Front 16

Common dyes

Back 16

methylene blue, crystal violet, basic fuchsin

Front 17

Why do common dyes work?

Back 17

They have chromophores with positive charges and bacteria are negatively charged

Front 18

Two types of dyes

Back 18

Basic and Acidic

Front 19

- have positive (+) charge
- ex. Methylene blue
- can penetrate bacteria because of the attraction between charges

Back 19

Basic dyes

Front 20

- have negative charge
- ex. Eosin, Nigrosin
- can't penetrate bacteria because of repulsion between the charges
- stains background

Back 20

Acidic dyes

Front 21

- stains the background of the slide and not the organism
- useful for determining
cell shape and size
- these 2 cultures used in this test:
- no heat fixing means to cell distortion

Back 21

Negative staining; K.pneumoniae, S.epidermidis

Front 22

2 staining techniques used for capsular staining

Back 22

Negative and Simple

Front 23

Capsular staining technique uses ________ to color slide background and uses ___________ to color cell proper of the bacteria

Back 23

Congo Red; Maneval's stain

Front 24

The _________ is the defining part of whether the bacteria is Gram-negative or Gram-positive. The _____________ is based on the composition/appearance of [it]

Back 24

Cell wall; Differentiation

Front 25

The major element of the cell wall in bacteria is ___________

Back 25

Peptidoglycan

Front 26

For Gram-positive bacteria, the cell wall is mostly __% peptidoglycan; For Gram-positive bacteria, the cell wall is only ___% peptidoglycan

Back 26

~90%; ~10%

Front 27

- most Gram-positive walls have _______________ attached to them, which are partly responsible for the negative charge of the cell

Back 27

teichoic acids

Front 28

Gram-negative bacteria have a second layer besides the peptidoglycan layer called the _________________ because it is made of lipids and a variety of sugars

Back 28

Outer membrane (Lipopolysaccharide layer)

Front 29

In both types of cell wall, _________ forms an insoluble compound with _____________. A thick peptidoglycan layer prevents ______________ because the alcohol causes dehydration and shrinks the cell wall, so the dye becomes trapped

Back 29

Iodine; Crystal violet; Decolorization

Front 30

This is a differential stain used to detect the presence of mycolic acid

Back 30

Acid-Fast

Front 31

- rod-shaped, Gram-positive bacteria which like to grow at ~30 degrees C
- have unique waxy lipid layer called _____________ which makes the colonies look waxy and crusty
- resistant to dehydration

Back 31

Mycobacterium; Mycolic acid

Front 32

food that is used for culturing bacteria and other microorganisms

Back 32

Media

Front 33

Three consistencies of Media

Back 33

1) Liquid
2) Solid
3) Semisolid

Front 34

A good solidifying agent:

Back 34

1) is not utilized by the microorganisms
2) doesn't inhibit bacterial growth
3) Doesn't liquify at room temperature

Front 35

This is sometimes used as a solidifying agent, but it is hydrolyzed by many organisms and liquifies at room temp

Back 35

Gelatin

Front 36

Two good choices for solidifying agents

Back 36

Agar and silica gel

Front 37

chemical compounds that can be broken down into or used to make monomers

Back 37

Nutrients

Front 38

nutrients required in large amounts: ex. C, N, P, S, K, Mg, Ca, Na, Fe

Back 38

Macronutrients

Front 39

nutrients required in small amounts: ex. metals

Back 39

Micronutrients

Front 40

7 Nutritional requirements for media preparation:

Back 40

1) water
2) carbon
3) energy
4) nitrogen
5) minerals
6) growth factors
7) pH

Front 41

Media that has been synthesized; has exact composition; compounds used are highly purified and defined

Back 41

Synthetic or defined Media

Front 42

Media where exact composition not known; utilizes compounds that include all nutrients but aren't defined

Back 42

Non-synthetic or complex Media

Front 43

4 Types of Special Media

Back 43

1) Selective
2) Differential
3) Minimal
4) Enriched

Front 44

Media that contains the basic requirements for the bacteria to grow such as salts, a carbon source, and nitrogen

Back 44

Minimal Media

Front 45

media that contains all basic requirements but adds amino acids, vitamins, and minerals

Back 45

Enriched Media

Front 46

media that allows only certain organisms to grow; selection based on absence of certain nutrients or presence of inhibitory substance

Back 46

Selective Media

Front 47

media that contains substances which change the appearance of one species allowing for differentiation; ex. EMB-Eosin Methylene Blue

Back 47

Differential Media

Front 48

an acidic dye which causes coliforms (bacteria found in the intestine) to turn a metallic green color

Back 48

eosin

Front 49

the killing of bacteria or endospores found in media or glassware; can be perfomed using an _____________

Back 49

Sterilization; Autoclave

Front 50

Minimum requirements for Sterilization

Back 50

For 1 liter of Media, 15 minutes at 121.6 degrees C (250 degrees F) and 15 psi (pounds per square inch

Front 51

2 methods for obtaining pure culture:

Back 51

Streak plate and Pour plate

Front 52

Streak plate has 4 techniques:

Back 52

1) Quadrant streak
2) Radiant streak
3) 'T' streak
4) Continuous streak

Front 53

Methods uses plates and agar tubes to isolate loopfuls of bacteria; requires less skill and practice but a lot of media and supplies

Back 53

Pour plate

Front 54

When grown at 25-30 degrees C, this organism produces a red pigment called ____________. When grown at higher temperatures, it appears _________ on the media. It is found all over the environment. What is this organism?

Back 54

Prodigiosin; white; Serratia Marcescens

Front 55

This organism has endotoxins which cause disease like food poisoning. It is found all over the environment, and is cream colored. What is this organism?

Back 55

Escherichia coli

Front 56

This organism is a common laboratory contaminant, is found all over environment, and yellow colored due to carotenoid pigments. What is this organism?

Back 56

Micrococcus luteus

Front 57

This Agar is used to isolate and differentiate pathogenic organisms like Staphylococcus aureus. It is a selective and differential media

Back 57

Mannitol Salt Agar (MSA)

Front 58

This Agar is used to isolate Streptococci and Staphylococci from mixed cultures. It is a selective media. It selects for Gram positive bacteria and inhibits the growth of Gram negative bacteria

Back 58

Phenyl Ethyl Alcohol (PEA)

Front 59

This agar used to isolate fecal coliforms. It is a selective and differential media. Its sugars provide fermentable substrates that promote the growth of fecal coliforms

Back 59

Eosin Methylene Blue (EMB)

Front 60

This agar used to select and differentiate a member of Enterobacteriaceae; a selective and differential media; contains lactose, bile salts, neutral red, and crystal violet

Back 60

MacConkey Agar

Front 61

Consists of yeasts and molds; belong to Kingdom Myceteae; Mycology is the study of this

Back 61

Fungi

Front 62

Type of mold that has mass of intermeshed hyphae; what is seen microscopically

Back 62

Mycelium

Front 63

microscopic filaments that may or may not have walls

Back 63

Hyphae

Front 64

Has pseudohyphae which are multicellular structures produced through budding; has spores, which are smaller in size than parent cell and produced by budding

Back 64

Yeast

Front 65

viruses that infect bacteria; _________ that paratisize E.Coli are referred to as ______________

Back 65

phages; coliphages

Front 66

2 types of viruses

Back 66

Lytic and Lysogenic

Front 67

type of virus considered virulent; enters cell, takes over, replicates, then lyses cell for release

Back 67

Lytic

Front 68

type of virus considered temperate; enters cell, integrates into DNA of host; cell replicates normally then eventually viruses replicate and lyse cell to be released

Back 68

Lysogenic

Front 69

Viruses are observed with __________ microscope; they're quantified by their effect on host cells so you count the ___________

Back 69

electron; VIUs (virus infection unit)

Front 70

3 steps to isolate bacteriophage from sewage

Back 70

1) Enrichment (performed in rows)
2) Filtration (rows)
3) Separation (pairs)

Front 71

This type of isolation increases number of phage in raw sewage; performed by combining ___________ (source of phage), __________ (host of phage), and __________ (DSPB). The DSPB increases the host #, which increases the source #.

Back 71

Enrichment; sewage; E.Coli; Deca Strength Phage Broth

Front 72

This type of isolation separates the E.Coli from the phage; uses a ___________ filter which allows the virus to pass but not bacteria; sample must be ______________ to rid all cellular debris so membrane isn't clogged; use ______ plates, 3 per filtration group

Back 72

Filtration; 0.2 nanometer pore size; centrifuged; EMB (Eosin Methylene Blue)

Front 73

This type of isolation is used to find evidence of phage and to find the concentration of phage in original filtrate

Back 73

Seeding

Front 74

an organism's ability, or lack thereof, to grow in the presence of oxygen

Back 74

Aerotolerance

Front 75

5 Oxygen classes of bacteria:
1) requires oxygen, uses it as final electron acceptor
2) grow in the presence or absence of oxygen, but prefer oxygen
3) cannot grow in presence of oxygen
4) requires free oxygen, but only in limited amounts
5) microaerophile that requires elevated levels of carbon dioxide to survive

Back 75

1) Aerobe
2) Facultative Anaerobe
3) Anaerobe
4) Microaerophiles
5) Capnophiles

Front 76

4 Types of Media for determining Oxygen requirements:

Back 76

1) TSA deep
2) TSA shake
3) Fluid Thioglycollate Medium (FTM)
4) TSA plate

Front 77

solid agar; more oxygen at the top of the agar to less or no oxygen at the bottom of the tube

Back 77

TSA deep

Front 78

Agar kept in liquid state by placing tube in warm waterbath. After inoculation, tube gently rolled between palms to disperse bacteria, then is allowed to solidify. As it solidifies, oxygen gradient is created

Back 78

TSA shake

Front 79

basic medium, oxygen gradient formed in media during cooling; contains ___________ and ___________, reducing agents that reduce oxygen present in media to water; has __________, a dye used as an oxygen indicator (pink, oxygen present) (colorless, oxygen not present); ______ and __________ present in small quantities to localize organisms

Back 79

Fluid Thioglycollate Medium (FTM); Thioglycollate, L-cystine; Resazurin; Agar, gelatin

Front 80

gas-generator envelope used for providing anaerobic environment; has sachet of __________ which acts as catalyst

Back 80

GasPak apparatus; palladium

Front 81

____________ tests for cytochrome, part of electron transport chain. It has positive (___________) and negative (____________) control; Positive result: _____________ color change within 15 seconds. Negative result: ________________ within 15 seconds

Back 81

Oxidative Test; P.aeruginosa; E.Coli; Dark blue (purple); no color change

Front 82

Sugar Fermentation Tubes test for fermentation of 4 sugars:

Back 82

1) Glucose
2) Lactose
3) Mannitol
4) Sucrose

Front 83

S.F.T's reagent: pH indicator is _____________, detects acid production among fermentation; pH of media will drop as acid is produced; (_______ for pH}7 and __________ for pH{7)

Back 83

Phenol Red; Red; Yellow

Front 84

Results for S.F.T's: If positive, ________ color change and ___________ in tube for gas production. If negative, _________ color change and ___________ in tube for no gas production

Back 84

Yellow; Bubble; Red; No Bubble

Front 85

_______________tests for presence of nitrate and presence of nitratase. During test, add 15 drops of each reagent to test and determine color change, if none, add _________. Results: positive if ____ color change or ____ color change once ______ is added; negative if ___ color change initially once _______ is added and causes red color change

Back 85

Nitrase Test; Zinc;Red, no, zinc; no, zinc

Front 86

___________ looks for presence of emzyme catalase which breaks down toxic oxygen compounds into water and oxygen. Reagent: _______________. Performed using TSA slant. Positive result: ___________. Negative result: ___________.

Back 86

Catalase Test; Hydrogen peroxide; Bubbles; No Bubbles

Front 87

___________ (MR Test) tests for production of acids upon glucose fermentation. Reagent: ___________. pH indicator: red for pH _______, yellow for pH ________. Positive: ______. Negative: _________

Back 87

Methyl Red; Methyl Red; {5; }5; Red; Yellow

Front 88

___________ (VP Test) tests for production of alcohol; Look for color change: positive, __________; negative, _________

Back 88

Voges-Proskauer; Barritt's reagents; Red; Yellow

Front 89

______________ tests for unknown bacteria and utilization of citrate. Positive: ___________ color. Negative: ___________ color.

Back 89

Simmons Citrate Test; Prussian Blue; Green

Front 90

______________ tests for unknown bacteria and presence of phenylalanase. Positive: ____________ color. Negative: ___________ color.

Back 90

Phenylalanine Deanimation Test; Deep Green; Yellow

Front 91

_________ tests for unknown bacteria and presence of H2S production; semi-solid media; Positive: __________, cloudy tube, spreads throughout, dark precipitate. Negative: growth only on _____________, no dark precipitate

Back 91

SIM Media Deep

Front 92

___________ tests for unknown bacteria and presence of H2S production, fermentation of glucose, gas production. Positive: ____________, cracks in media, dark precipitate; Glucose if _____ color, ______ at bottom of tube. Negative: no cracks in media, no ______________, glucose if _____ in bottom of tube, lactose if _______ throughout tube.

Back 92

Kilgar's Iron Agar (KIA); gas production; red, yellow; dark precipitate, red, red

Front 93

Hydrolysis Test, looks for urease: reagent Phenol Red; Positive: _________, Negative: _______

Back 93

Urea Hydrolysis; Red, Yellow

Front 94

Hydrolysis Test, looks for tryptophanase: Positive if ________ at top, negative if _______ at top

Back 94

Tryptone Hydrolysis; red ring; no red ring

Front 95

Hydrolysis Test, looks for lipases; media is _________; positive _____________; negative _____________.

Back 95

Fat Hydrolysis (SBA); Spirit Blue Agar; Dark Precipitate; no dark precipitate

Front 96

Hydrolysis Test, looks for presence of caseinase; Positive, ________ around growth; negative, _________

Back 96

Casein Hydrolysis (Milk Agar); clear zone around growth, no zone round growth

Front 97

Hydrolysis Test, tests for amylases. Positive, _____________. Negative, ___________

Back 97

Starch Hydrolysis; clear zone around growth, no zone round growth, media brownish

Front 98

UV light divided into 3 groups:

Back 98

1) UV-A, longest wave, 315 to 400
2) UV-B, wave between 280 to 315
3) UV-C, wave from 100 to 280 (most detrimental to bacteria)

Front 99

Which one is most detrimental to bacteria? UV causes death by causing DNA damage in the form of _______________

Back 99

UV-C; pyrimidine dimers

Front 100

This tests effectiveness of antibiotics on bacteria. It utilizes this agar to meets its standard requirements. If inoculated bacterium is susceptible to antibiotic, it will produce _________________. Size of it depends on bacteria sensitivity to __________________

Back 100

Kirby-Bauer Method; Mueller Hinton Agar; zone of inhibition; Minimum Inhibitory Concentration

Front 101

Kirby-Bauer Method tests against following bacteria:

Back 101

1) E.Coli
2) Proteus vulgaris
3) Pseudomonas aeruginosa
4) Staphylococcus aureus

Front 102

During Kirby-Bauer Method, ________ and __________ will be white or cream-colored lawn; _________ will be yellow; __________ will be green

Back 102

E.Coli and P.Vulgaris; P.aeruginosa; S.aureus

Front 103

3 major Staphylococcus pathogens:

Back 103

S.auerus, S.epidermis, S.saprophyticus

Front 104

- most pathogenic of Staphylococcus
- can cause Toxic Shock Syndrome
- definitive test is ___________
- color is yellow, positive in everything
- This tests for its identification
- has 2 proteins: __________ and __________

Back 104

S.aureus; Coagulase Plasma; Bacterial Agglutination; coagulase, protein A

Front 105

- color is white
- negative in everything
- most likely won't ferment mannitol

Back 105

S.epidermis

Front 106

- organism whose color is white
- negative in everything except mannitol fermentation

Back 106

S.saprophyticus

Front 107

-selective media
- contains mannitol, 7.5% NaCl
- favors colony pigmentation

Back 107

Staphylococcus Medium (SM110)

Front 108

3 major types of hemolysis

Back 108

1) alpha - partial lysis of red blood cells to discolorization to green
2) beta - complete lysis results in clear zone
3) y - no lysis

Front 109

- cocci shaped
- forms chains of organisms when growing
- can cause meningitis, pneunomia

Back 109

Streptococci

Front 110

3 medically significant species of Streptococci

Back 110

1) S.pyrogenes - strep throat, Scarlet fever
2) S.agalactiae
3) S.pneumoniae - pneumonia, memningitis

Front 111

7 major groups of Streptococci

Back 111

A-D enterococci, D non-enterococci, viridians, pneumococci

Front 112

3 tests for alpha-hemolytic Strep.

Back 112

1) Bile Esculin Agar slant (6ml)
2) 6.5% NaCl Tolerance Broth (5ml)
3) Optochin Sensitivity

Front 113

3 tests for beta-hemolytic Strep.

Back 113

1) CAMP Reaction
2) Bacitracin Susceptability
3) SXT Susceptability

Front 114

This agar differentiates Strep species. It contains _______, which provides nutrients, _______, which inhibits growth of Gram-positive organisms, and _________, an indicator. It indicates Strep belongs to Group ____: has 2 subgroups, ________ (S.faecalis) and _______ (S.bovis)

Back 114

Bile Esculin Agar Slant (6ml); Bile; Esculin; Ferric citrate; D; D enterococci, D-nonenterococci

Front 115

tests ability of some Strep. to grow in 6.5% NaCl. Its pH indicator is the color ____________, and if this color changes to yellow a ________ result occurs; it differentiates Group ___ from Group ____

Back 115

6.5% NaCl Tolerance Broth; purple; positive; D; non-D

Front 116

This test differentiates pneumococci from viridians; positive test indicated by zone of inhibition

Back 116

Optochin Susceptability

Front 117

This test is positive by indication of formation of arrowhead; indicates Strep belongs to group ___

Back 117

CAMP Reaction; B

Front 118

This test has 2 limitations, positive result by any zone of inhibition; mostly indicates Strep belongs to group ___, but 5% group ___ and 10% group ___

Back 118

Bacitracin Susceptability; A, B, C

Front 119

Group A and group B are resistant to this test. It distinguishes these groups from non-A and non-B groups

Back 119

SXT Susceptability

Front 120

anaerobic process in which sugar is converted to alcohol by CO2 and yeast (_______________)

Back 120

Fermentation; Saccharomyces cervisiae

Front 121

C6H12O6 --through yeast-- 2C2H5OH + 2CO2

Back 121

Reaction of Fermentation

Front 122

Group A and group B are resistant to this test. It distinguishes these groups from non-A and non-B groups

Back 122

SXT Susceptability

Front 123

anaerobic process in which sugar is converted to alcohol by CO2 and yeast (_______________)

Back 123

Fermentation; Saccharomyces cervisiae

Front 124

C6H12O6 --through yeast-- 2C2H5OH + 2CO2

Back 124

Reaction of Fermentation

Front 125

4 types of flagellar arrangements:

Back 125

1) amphitrichous (at both ends)
2) Lephotrichous (3 at one end)
3) Monotrichous (single at one end)
4) Peritrichous (all around perimeter)

Front 126

3 types of movement in bacteria

Back 126

1) gliding
2) rotary
3) flagellar

Front 127

this genus produce endospores

Back 127

Bacillus (aerobes) and Cloistridium (anarobes)

Front 128

in spore staining, ___________ is primary stain, ____________ is counterstain

Back 128

Malachite green, Safranin