Lab Practical Ii

Front 1

Effects of extreme temperatures on living

organisms

Back 1

Cold= slows activity

Hot= denaturation

Front 2

Psychrophiles

Back 2

optimal temp= -5*C - 20*C

Front 3

Mesophiles

Back 3

optimal temp= 20*C - 45*C

(human pathogens)

Front 4

Thermophiles

Back 4

optimal temp= 35*C and above

Front 5

Facultative Thermophiles

Back 5

will grow at 37*C with optimum growth

45*C - 60*C

Front 6

Obligate Thermophiles

Back 6

only grow at temps above 50*C, prefer temps above 60*C

Front 7

Acidophiles

Back 7

0-7 pH, optimal is 3.5

Front 8

Neutrophiles

Back 8

3.5-10.5 pH, optimal is 7

(human pathogens)

Front 9

Alkalophiles

Back 9

7-14 pH, optimal is 10.5

Front 10

Aerobes

Back 10

*require O2 for growth

*growth on top of agar

Front 11

Microaerophiles

Back 11

*Require minimal amounts of O2 for growth, too much O2 results in death.

*Growth just under surface of agar

Front 12

Obligate Anaerobes

Back 12

*Require the absence of O2 for growth.

*growth on bottom of agar

Front 13

Aerotolerant anaerobes

Back 13

*Do not require O2, won't kill but cannot use it

*Growth evenly throughout agar

Front 14

Facultative Anaerobes

Back 14

*Can grow in the presence or absence of O2.

*Growth throughout agar with more on surface

Front 15

Method/Procedure of Classifying

O2 Requirements

Back 15

Shake-tube Inoculation in melted agar followed by rapid solidification.

Front 16

What does lifting of agar mean after shake tube inoculation?

Back 16

*Presence of CO2 (fermentation)

*Obligate anaerobes

Front 17

2 Methods used to cultivate strict anaerobes

Back 17

1. gas pak

2. thioglycollate broth

Front 18

Action of Resazurin in thioglycollate broth

Back 18

Resazurin= redox potential indicator

clear= presence of O2

colorless= absence of O2

Front 19

What is the effect of UV radiation on DNA?

Back 19

thymine dimerization

Front 20

Is UV radiation ionizing or non-ionizing?

Back 20

Non-ionizing

Front 21

Can UV radiation penetrate petri dish lid?

Back 21

No

Front 22

Advantages of Serial Dilution

Back 22

1. only viable cells are counted

2. allows isolation of colonies for subculturing into pure cultures.

Front 23

Disadvantages of Serial Dilution

Back 23

1. Overnight Incubation

2. Use of more glassware

3. May result in errors due to large amount of steps.

Front 24

Why must molten agar be cooled before pouring the plates?

Back 24

So as not to kill the organism

Front 25

What is the purpose of the "Figure 8" action?

Back 25

To mix cells into agar.

Front 26

TNTC

Back 26

Too Numerous To Count= over 300

Front 27

TFTC

Back 27

Too Few To Count= under 30

Front 28

definition of antibiotic

Back 28

Chemotherapeutic agent synthesized and

secreted by some bacteria, actinomycetes, and fungi that destroy or inhibit the growth of other microorganisms.

Front 29

Kirby-Bauer Procedure

Back 29

A diffusion procedure used to determine drug succeptibility by measuring the zone of inhibition resulting from diffusion.

Front 30

How are forceps are disinfected during Kirby-Bauer procedure?

Back 30

Dipped in alcohol and flamed between antibiotic disks.

Front 31

Describe antibiotic sensitivity on agar plate:

Back 31

Presence of zone of inhibition around disk.

Front 32

Describe antibiotic resistance on agar plate:

Back 32

Absence of zone of inhibition around disk.

Front 33

Type of agar used for antibiotic sensitivity test:

Back 33

Mueller-Hinton agar plates

Front 34

Describe synergistic effect of antibiotic

combinations as seen on agar:

Back 34

Zones of inhibition of disks merge together.

Front 35

Describe additive effect of antibiotic

combinations as seen on agar:

Back 35

Zones of inhibition are separate and do not merge.

Front 36

definition of Sterilize:

Back 36

to remove all life

Front 37

definition of Disinfect:

Back 37

reduction of living organisms

Front 38

definition of Microbistatic:

Back 38

to inhibit growth of microbes

Front 39

definition of Microbicidal:

Back 39

to kill microbes

Front 40

Factors that can inhibit effectiveness of a

disinfectant/antiseptic solution:

Back 40

1. Concentration

2. Length of Exposure

3. Type of microbial population to be destroyed

4. Environmental Conditions

a. temperature

b. pH

c. type of material on which the

organism exists

Front 41

Describe effect of disinfectant/antiseptic

sensitivity on agar plate:

Back 41

Zone of inhibition is indicative of

disinfectant/antiseptic sensitivity

Front 42

Disinfectants/Antiseptics used in lab:

Back 42

1. iodine

2. hydrogen peroxide

3. isopropyl alcohol

4. chlorine bleach

5. Lysol

6. Listerine

Front 43

To what Kingdom to Protozoa belong to?

Back 43

Kingdom Protista

Front 44

Method of Classification for Protozoans:

Back 44

Mode of locomotion

Front 45

Mode of Locomotion for Paramecium caudatum:

Back 45

Cilia

Front 46

Mode of Locomotion for Amoeba proteus:

Back 46

pseudopods

Front 47

Mode of Locomotion for Balantidium coli:

Back 47

cilia

Front 48

Mode of Locomotion for Trypanosoma

gambiense:

Back 48

Flagella

Front 49

Mode of Locomotion for Plasmodium vivax:

Back 49

None

Front 50

To which class does B. coli belong?

Back 50

Class Ciliophora

Front 51

To which class does Amoeba proteus belong?

Back 51

Class Sarcodina

Front 52

To which class does Trypansoma belong?

Back 52

Class Mastigophora

Front 53

To which class does Plasmodium belong?

Back 53

Class Sporozoa/ Apicomplexa

Front 54

To which Kingdom do Helminths belong?

Back 54

Kingdom Animalia

Front 55

Disease caused by Amoeba:

Back 55

dysentery

Front 56

Disease caused by B. coli

Back 56

dysentery

Front 57

Disease caused by Trypansoma

Back 57

African Sleeping Sickness

Front 58

Disease caused by Plasmodium

Back 58

Malaria

Front 59

Disease caused by tapeworms

Back 59

Taenia solium

Front 60

Disease caused by flukes

Back 60

Fasciola hepatica

Front 61

Disease caused by roundworm Trichanella

Back 61

Trichinosis

Front 62

def. of hermaphroditic:

Back 62

an organism that has working reproductive

organs normally associated with both male and female sexes

Front 63

What Helminths are hermaphroditic?

Back 63

Tapeworms

Flukes

Front 64

Carbohydrate fermentation test

Purpose:

Back 64

to determine the ability of a

microorganism to degrade and ferment carbohydrates with the presence

Front 65

Carbohydrate fermentation test

Medium:

Back 65

Carbohydrate fermentation broth with Durham tube

Front 66

Carbohydrate fermentation test

Substrates:

Back 66

lactose, sucrose dextrose

Front 67

Carbohydrate fermentation test

End Products:

Back 67

lactic acid or,

lactic acid & gas

Front 68

Carbohydrate fermentation test

pH indicator:

Back 68

phenol red in medium

Front 69

Carbohydrate fermentation test

Results:

Back 69

Neg. result: no color change (red), no gas

Pos. result: color change to yellow/gas

Front 70

Indole Production Test (IMViC)

Purpose:

Back 70

to determine the ability of

microorganisms to degrade tryptophan

Front 71

Indole Production Test (IMViC)

Medium:

Back 71

SIM agar deep

Front 72

Indole Production Test (IMViC)

Substrate:

Back 72

tryptophan

Front 73

Indole Production Test (IMViC)

Enzyme:

Back 73

tryptophanase

Front 74

Indole Production Test (IMViC)

End Products:

Back 74

Indole

Front 75

Indole Production Test (IMViC)

pH indicator:

Back 75

Kovac's 10 drops

Front 76

Indole Production Test (IMViC)

Results:

Back 76

Neg. result: brown reagent layer

Pos. result: red reagent layer

Front 77

Methyl Red Test

Purpose:

Back 77

to determine the ability of

microorganisms to ferment glucose with the production and stabilization of high

concentrations of acid end production.

Front 78

Methyl Red Test

Medium:

Back 78

MR-VP broth

Front 79

Methyl Red Test

Substrate:

Back 79

Glucose

Front 80

Methyl Red Test

End Product:

Back 80

Acid

Front 81

Methyl Red Test

pH indicator/reagent:

Back 81

Methyl Red 10 drops

Front 82

Methyl Red Test

Results

Back 82

Neg. result: yellow

Pos. result: red

Front 83

Vogues-Proskauer Test

Purpose:

Back 83

differentiate further among enteric organisms (E.coli, E.aerogenes, K.pneumoniae)

Front 84

Vogues-Proskauer Test

Medium:

Back 84

MR-VP broth

Front 85

Vogues-Proskauer Test

Substrate:

Back 85

Glucose

Front 86

Vogues-Proskauer Test

End Product:

Back 86

neutral/non acidic

Front 87

Vogues-Proskauer Test

pH indicator/reagent:

Back 87

VP-A & VP-B 10 drops

Front 88

Vogues-Proskauer Test

Results:

Back 88

Neg. result: clear, no color change

Pos. result: Red

Front 89

Citrate Utilization Test

Purpose:

Back 89

differentiate among enteric

organisms on the basis of ability to ferment citrate as a sole source of carbon.

Front 90

Citrate Utilization Test

Medium:

Back 90

Simmon's citrate agar slant

Front 91

Citrate Utilization Test

Substrate:

Back 91

Citrate

Front 92

Citrate Utilization Test

Enzymes:

Back 92

Citrate Permease and Citrase

Front 93

Citrate Utilization Test

End Product:

Back 93

Sodium Carbonate (alkaline)

Front 94

Citrate Utilization Test

pH indicator/reagent:

Back 94

Bromythmol blue

Front 95

Citrate Utilization Test

Results:

Back 95

Neg. result: no color change, green

Pos. result: Royal blue color

Front 96

Hydrogen Sulfide Test

Purpose:

Back 96

Motility and ability of

microorganisms to produce hydrogen sulfide from sulfur-containing amino acids or

inorganic sulfur compounds

Front 97

Hydrogen Sulfide Test

Medium:

Back 97

SIM agar deep

Front 98

Hydrogen Sulfide Test

Substrate:

Back 98

Cysteine, Thiosulfide

Front 99

Hydrogen Sulfide Test

Enzymes:

Back 99

Cysteine Desulferase, Thiosulfate

Reductase

Front 100

Hydrogen Sulfide Test

End Product:

Back 100

Hydrogen Sulfide gas+ Ferrous Ions = FeS

Front 101

Hydrogen Sulfide Test

Results:

Back 101

Neg. result: No black, no motility

Pos. result: Black, motility= growth away from stab line

Front 102

Urease Test

Purpose:

Back 102

determine ability of microorganism to degrade urea by means of enzyme urease

Front 103

Urease Test

Medium:

Back 103

Urea broth

Front 104

Urease Test

Substrate:

Back 104

Urea

Front 105

Urease Test

Enzyme:

Back 105

Urease

Front 106

Urease Test

End Product:

Back 106

Ammonia

Front 107

Urease Test

pH indicator/reagent:

Back 107

Phenol red

Front 108

Urease Test

Results:

Back 108

Neg. result: no color change

Pos. result: hot pink color

Front 109

Nitrate Reduction Test

Purpose:

Back 109

determine ability of microorganisms to reduce nitrates to nitrites or beyond the

nitrite stage.

Front 110

Nitrate Reduction Test

Medium:

Back 110

Nitrate reduction broth w/ Durham tube

Front 111

Nitrate Reduction Test

Substrate:

Back 111

Nitrate, Nitrite

Front 112

Nitrate Reduction Test

Enzymes:

Back 112

Nitrate Reductase

Front 113

Nitrate Reduction Test

End Product:

Back 113

Nitrite, N2, or NH3

Front 114

Nitrate Reduction Test

pH indicator/reagent:

Back 114

Nitrate A & B, Zinc

Front 115

Nitrate Reduction Test

Results:

Back 115

Neg. result for nitrate=No color change, red

for nitrite= Red

Pos. result for nitrate= Red

for nitrite= unchanged

Front 116

Catalase Test

Purpose:

Back 116

determine ability of microorganisms to degrade hydrogen peroxide by producing catalase.

Front 117

Catalase Test

Medium:

Back 117

TSA slant

Front 118

Catalase Test

Substrate:

Back 118

Hydrogen peroxide

Front 119

Catalase Test

End Product:

Back 119

Oxygen

Front 120

Catalase Test

pH indicator/reagent:

Back 120

Hydrogen Peroxide, 3 drops

Front 121

Catalase Test

Enzyme:

Back 121

Catalase

Front 122

Catalase Test

Results:

Back 122

Neg. result: no bubbles

Pos. result: bubbles

Front 123

Steps of Unknowns:

Back 123

1. Streak plate= to isolate pure colonies

2. Agar slant= to get pure culture

3. Gram-stain= to prove purity

4. Broth subculture= for fresh nutrients

5. Biochemical tests= to identify species

Back 124

Paramecium caudatum

Back 125

Amoeba proteus

Back 126

Balantidium coli (trophozoite)

Back 127

Balantidium coli (cyst)

Back 128

Trypanosoma gambiense

Back 129

Plasmodium vivax

Back 130

*Tapeworm

*Taenia solium

*Scolex (head region)

Back 131

Tapeworm

Proglottids

Back 132

Fasciola Hepatica

Back 133

Roundworm

*Ascaris lumbricoides

Left=male

Right=female

Back 134

Trichinella spiralis