Dvc Bloom Bio 146 Lab Practical 1

Front 1

Back 1

Arm

Front 2

Back 2

Base

Front 3

Back 3

Condenser

Front 4

Back 4

Course and Fine Focus Knobs

Front 5

Back 5

Iris diaphragm

Front 6

Back 6

Light Rheostat and should be set at 4.5

Front 7

Back 7

Light Source

Front 8

Back 8

Light Switch

Front 9

Back 9

Objective Lenses

Front 10

Back 10

Ocular Lenses (10x)

Front 11

Back 11

Stage with Stage Clips

Front 12

What power objective lens should you start with for all bacterial slides?

Back 12

10x

Front 13

What power objective lens should you start with for all eukaryotic slides?

Back 13

4x

Front 14

Should you always view bacteria slides in oil immersion?

Back 14

yes the 100x objective lens

Front 15

Define total magnification.

Back 15

Objective lens x Ocular lens (10x)

Front 16

What is resolution?

Back 16

is the amount of detail you can see in an image.

Front 17

What is the Sterile Technique?

Back 17

allows for the transfer of organisms from one place to another without contamination.

Front 18

What is a pure culture?

Back 18

Means that only one type of organism is present.

Front 19

Define aseptic technique?

Back 19

vital in hospital settings in preventing cross contamination; flame sterilization of all instruments and tools all the time.

Front 20

What percentage of hospitalized patients acquire healthcare associated infections?

Back 20

5 to 15%

Front 21

What is
nosocomial infections

Back 21

hospital acquired infections

Front 22

Why do we perform the streak plate technique?

Back 22

to produce isolated colonies of one species of bacteria

Front 23

What is this?

Back 23

pond water (the algae help identify it since it shows up green)

Front 24

Which organisms in pond water are eukaryotic?

Back 24

Algae and protozoa

Front 25

why is fluorescence microscopy is used

Back 25

provides contrast to see specimen glowing a color against a dark background

Front 26

Identify the types of cells

Back 26

Rod & Coccus

Front 27

What type of staining technique is used?

Back 27

Gram stain

Front 28

What type of staining technique is used?

Back 28

Leifson Flagella stain

Front 29

What type of staining technique is used?

Back 29

Capsule Stain

Front 30

What type of staining technique is used?

Back 30

Schaeffer-Fulton endospore stain

Front 31

what type of staining technique is used?

Back 31

Ziehl-Neelsen stain

Front 32

What type of stain is the Gram stain?

Back 32

A differential stain, 1st stain of choice in identifying an unknown.

Front 33

What are the steps of a Gram Stain

Back 33

Heat fix smear
Primary stain is applied
mordant is added
decolorized with ethanol
counterstain (safranin)
Wash

Front 34

What type of mordant is used in the gram stain?

Back 34

Iodine

Front 35

What is the primary stain used in the Gram Stain?

Back 35

Crystal Violet

Front 36

What is the counterstain used in the Gram Stain?

Back 36

Safranin (red)

Front 37

Name four secondary stains

Back 37

Acid-fast
Endospore
Flagella
Capsule

Front 38

What type of technique is used in the Acid Fast Stain?

Back 38

Ziehl Neelsen

Front 39

What is the primary stain used in the Ziehl Neelsen technique?

Back 39

Carbolfuschin (pink)

Front 40

What is the counterstain used in the Ziehl Neelsen technique?

Back 40

Methylene blue

Front 41

Which two techniques are used for endospore stains?

Back 41

1 - crystal violet method
2 - schaeffer-fulton method

Front 42

Describe the crystal violet method

Back 42

negative staining technique
stains vegetative cells highlighting endospores in purple

Front 43

describe the shaeffer fulton method

Back 43

uses a primary stain malachite green to stain endospores
rinse
counter stain safranin

Front 44

What technique is used for the flagella stain

Back 44

Leifson's flagella

Front 45

Describe Leifson's flagella

Back 45

rolling drop slide
air dried
stain (contains a mordant) applied
-inflate flagella

Front 46

What is the special external protective layer that makes viewing capsules possible?

Back 46

defined by the form of glycocalyx; an external outer protective layer

Front 47

Define the capsule stain

Back 47

negative stain
uses India Ink to stain background
Counterstain Safranin to stain bacterial cells
air dried; heat fixing gives false results

Front 48

What is brownian motion?

Back 48

vibration effect

Front 49

how do you determine motility?

Back 49

Using a hanging drop technique
purposeful forward swimming

Front 50

Motility: Is it motile and what is it's shape?
Bacillus subtilis

Back 50

M+, big rod

Front 51

Motility: Is it motile and what is it's shape?
Enterobacter aerogenes

Back 51

M+, rod

Front 52

Motility: Is it motile and what is it's shape?
Klebsiella pnemoniae

Back 52

M-, rod

Front 53

Motility: Is it motile and what is it's shape?
Micrococcus luteus

Back 53

M-, coccus

Front 54

Motility: Is it motile and what is it's shape?
Proteus Mirabilis

Back 54

M+, rod

Front 55

Motility: Is it motile and what is it's shape?
Rhodospirillum rubrum

Back 55

M+, rod

Front 56

Motility: Is it motile and what is it's shape?
Sporosarcina ureae

Back 56

M+, coccus

Front 57

Motility: Is it motile and what is it's shape?
Janthinobacterium lividum

Back 57

M+, rod

Front 58

What is the Gram Rxn & Morphology for:
Bacillus Subtilis (agar)

Back 58

G+, rod

Front 59

What is the Gram Rxn & Morphology for:
Bacillius Subtilis (broth)

Back 59

G+, rod

Front 60

What is the Gram Rxn & Morphology for:
E.Coli (agar)

Back 60

Gneg, rod

Front 61

What is the Gram Rxn & Morphology for:
E. Coli (broth)

Back 61

Gneg, rod

Front 62

Identify the bacteria, Gram reaction and morphology.

Back 62

Bacillus subtilis
G+
Rod

Front 63

Identify the bacteria, gram reaction, and morphology.

Back 63

E. Coli
Gneg
Rod

Front 64

Identify the bacterium based on colony morphology and it's optimal temperature.

Back 64

Bacillus Globigii
37*C

Front 65

Identify a bacterium based on colony morphology and its optimum temperature.

Back 65

Bacillus Licheniformis
37*C

Front 66

Identify a bacterium based on colony morphology and its optimum temperature.

Back 66

Bacillus Megaterium
Not much difference b/w temperatures.

Front 67

Identify a bacterium based on colony morphology and its optimum temperature.

Back 67

Bacillus Mycoides
22*C
Fungal/Mold like growth

Front 68

Identify the bacterium based on pigmentation and note any specifics re: temperature.

Back 68

Bacillus globigii
No noticeable differences b/w temperatures.

Front 69

Identify the bacterium based on pigmentation and note any specifics re: temperature.

Back 69

Chromobacterium violaceum
slightly darker at 22*C

Front 70

Identify the bacterium based on pigmentation and note any specifics re: temperature.

Back 70

Micrococcus luteus
bright yellow at both 22 and 37*C

Front 71

Identify the bacterium based on pigmentation and note any specifics re: temperature.

Back 71

Micrococcus roseus
salmon pink only grows at 22*C

Front 72

Identify the bacterium based on pigmentation and note any specifics re: temperature.

Back 72

Pseudomonas aeruginosa
opaque yellow at 22
watery blue at 37

Front 73

Identify the bacterium based on pigmentation and note any specifics re: temperature.

Back 73

Serratia marcescens
Red at 22*C

Front 74

Identify the bacterium based on pigmentation and note any specifics re: temperature.

Back 74

Janthinobacterium lividum
dark purple at 22*C
NG at 37*C

Front 75

Why do non-photosynthetic bacteria produce pigments?

Back 75

as byproducts of metabolism, as antioxidants, and as protection against solar radiation.

Front 76

Name the slime mold that we used in lab and the medium used.

Back 76

Physarum polycephalum sclerotia
Tap water agar plates (2%)

Front 77

Define cytoplasmic streaming.

Back 77

The flow of oxygen and nutrients from one part of the plasmodium to another in protoplasmic vessels.

Front 78

Define hanging drop slide

Back 78

suspends bacteria in a drop of fluid, reducing evaporation and water currents when compared to regular wet mounts.

Front 79

Define osmotic pressure

Back 79

indicates the solute concentration of the surrounding solutions and the effect it has on the flow of water across the membrane.

Front 80

What is the term to describe water leaving a cell that results in cell dehydration and shrinkage?

Back 80

plasmolysis

Front 81

Identify the test and the bacterium had the widest range of salt tolerance and explain why. Which bacterium is an example of an obligate halophile.

Back 81

Osmotic Pressure Test
Widest Range: Staphylococcus epidermidis (up to 15% NaCl) because it lives in salty human skin
Obligate Halophile: Halobacterium salinarium

Front 82

True or False: Bacillus stearothermophilus is an example of a thermophile.

Back 82

Bacillus stearothermophilus

Front 83

Name the five temperature classifications for organisms.

Back 83

Psychrophiles "cold loving"
Psychrotrophs
Mesophiles "middle loving"
Thermophiles "heat loving"
Hyperthermophiles/Extreme thermophiles

Front 84

What it the temperature range for psychrophiles?

Back 84

-5*C and <19/20*C

Front 85

What it the temperature range for psychrotrophs?

Back 85

-1*C and 35*C

Front 86

What it the temperature range for Mesophiles?

Back 86

12*C to 45*C
optimum 37*C

Front 87

What it the temperature range for thermophiles?

Back 87

41*C and 80*C

Front 88

What it the temperature range for Hyperthermophile?

Back 88

75*C and 112*C

Front 89

Which is room temperature:

37*C or 22*C

Back 89

22*C

Front 90

Which is body temperature:

37*C or 22*C

Back 90

37*C

Front 91

Define

bacteriostatic

Back 91

subsequent growth after initial incubation

Front 92

Define

bacteriocidal

Back 92

lack of growth after initial incubation

Front 93

What are two examples of bacteriocidal methods

Back 93

Cooking and boiling

Front 94

What are two examples of bacteriostatic methods

Back 94

Refrigerating and freezing

Front 95

What is the classification, optimum temp and temp range for
Bacillus licheniformis

Back 95

Mesophile
Optimum 37*C
Range: 12*C to 45*C

Front 96

What is the classification, optimum temp and temp range for
Bacillus stearothermophilus

Back 96

Thermophile
Optimum: 55*C
Range: 41*C and 80*C

Front 97

What is the classification, optimum temp and temp range for
E.Coli

Back 97

Mesophile
Optimum: 37*C
Range: 12*C to 45*C

Front 98

What is the classification, optimum temp and temp range for
Janithinobacterium lividum

Back 98

Psychotroph
Optimum 22*C
Range: -1*C and 35*C

Front 99

If a bacterium grows at a warmer temperature after incubating in a colder temperature - it is...

Back 99

bacteriostatic

Front 100

If a bacterium fails to grow at a warmer temperature after incubating in a colder temperature - it is...

Back 100

bacteriocidal

Front 101

What baceterium is an example of an anaerobe?

Back 101

Clostridum sporogenes

Front 102

Define obligate aerobes

Back 102

require oxygen, possess enzymes, catalase and superoxide dismutase (SOD)

Front 103

What is SOD

Back 103

superoxide dismutase

Front 104

What does catalase breakdown?

Back 104

hydrogen peroxide into O2 and H2O

Front 105

Define anaerobes

Back 105

cannot use oxygen in metabolic pathways

Front 106

Define
microaerophiles

Back 106

require only small amounts of O2 use aerobic respiration for energy

Front 107

Define
Obligate anaerobes

Back 107

only grow when no O2 is present; use fermentation or anaerobic respiration for energy

Front 108

Define
Facultative anaerobes

Back 108

can grow with or without O2; but grow best if O2 is present; possess enzymes, catalase and SOD

Front 109

Define
Aerotolerant anaerobes

Back 109

primarily fermenters; possess enzyme and SOD

Front 110

What does SOD breakdown?

Back 110

breaks down superoxide radical into hydrogen peroxide

Front 111

Which bacterium is an example of an obligate aerobe?

Back 111

Bacillus subtilis

Front 112

Which bacterium is an example of a strict anaerobe?

Back 112

Clostridium sporogenes

Front 113

Which bacterium is an example of a microaerophile and how can you tell?

Back 113

Lactococcus lactis
best growth in reduced O2 when compared to AER tubes

Front 114

Which bacteria are facultative anaerobes?

Back 114

Enterobacter aerogenes & E.Coli
They will grow most near the top but appear throughout the tube

Front 115

Which bacterium is an example of aerotolerant anaerobe?

Back 115

Lactobacillus casiae
will look as though growing from bottom up

Front 116

Describe how UV radiation controls bacterial growth.

Back 116

UV radiation kills microorganisms by damaging DNA. It creates thymine dimers that distort DNA, which lead to replication and transcription problems.

Front 117

Do bacteria have light and dark repair enzymes?

Back 117

Yes but these can be overwhelmed if exposure is significant.

Front 118

Which type of environments are most susceptible to UV radiation?

Back 118

Microrganisms in air, clear water, and on surfaces.

Front 119

How do hospitals use UV radiation?

Back 119

In germicidal lamps (ORs and nurseries).

Front 120

Differentiate a disinfectant from an antiseptic.

Back 120

Disinfectant is generally a chemical that kills or inhibits vegetative microbes. An ANTISEPTIC is a type of disinfectant that is nontoxic enough to be applies to skin and mucous membranes.

Front 121

Is this an antiseptic or disinfectant?
Isopropyl Alcohol (IA)

Back 121

Antiseptic

Front 122

Is this an antiseptic or disinfectant?
Hydrogen Peroxide (HP)

Back 122

Antiseptic

Front 123

Is this an antiseptic or disinfectant?
Lysol (Ly)

Back 123

Disinfectant

Front 124

Is this an antiseptic or disinfectant?
Bleach (Bl)

Back 124

Disinfectant

Front 125

What does a zone of inhibition indicate?

Back 125

antimicrobial activity

Front 126

You are looking at Bacillus Subtilis. Which chemical was LEAST effective and is it a disinfectant or an antiseptic?

Back 126

Isopropyl alcohol
antiseptic

Front 127

Identify this bacterium. Hint: it was the only one that had NG for HP and Bl and it is "topical".

Back 127

Staphylococcus epidermidis

Front 128

Which of these bacterial species is most UV resistant/tolerant and why?
Bacillus Subtilis, E. Coli, Staphylococcus epidermidis

Back 128

Bacillus subtilis; endospores resist UV;

Front 129

You are traveling when you suddenly need surgery. The village hospital only has liquid hand soap available for their doctors to scrub in. You have a small bottle of alcohol-based hand sanitizer in your day pack.

Do you...
A) go with the liquid based soap
B) insist on the alcohol based hand sanitizer

EXPLAIN WHY

Back 129

B - you insist on the alcohol-based hand sanitizer because
-it is more effective at killing transient organisms and agents
-it reduces the number of hospital acquired infections
-and you saw in lab that it killed 93% of bacteria present on your partner's hands.

Front 130

True or False:

Molds belong to the Kingdom Plantae?

Back 130

False. Molds belong to the Kingdom Fungi
Molds include mushrooms, mildew, and bread mold.

Front 131

True or False:

Yeasts belong to the Kingdom Bacteria?

Back 131

False. Yeasts belong to the Kingdom Fungi include unicellular nonmotile yeasts.

Front 132

Identify these life stages and identify type of mold.

Back 132

A. Spores
B. Spore germinating hypha
C. Hyphae going to Mycelium
Aspergillus Mold

Front 133

Name this mold.

Back 133

Rhizopus

Front 134

Name this mold.

Back 134

Penicillium

Front 135

What was the name of the selective medium used in the mold lab?

Back 135

Sabouraud Agar due to its low pH and high sugar concentration which inhibits bacterial growth

Front 136

What stain was used to make the molds visible?

Back 136

lactophenol cotton blue

Front 137

How did we collect the mold?

Back 137

Tape and toothpicks.

Front 138

What was the name of the yeast used in lab?

Back 138

Saccharomyces cerevisiae

Front 139

What are you looking at?

Back 139

Budding Yeast is most clearly present

Front 140

What is it?

Back 140

Yeast ascospore

Front 141

What method is used to estimate the number of bacteria in the soil.

Back 141

serial dilution of a sample of soil.

Front 142

Define serial dilution

Back 142

dilute one gram of sample soil through a series so that bacterial numbers are progressively decreased

Front 143

Why, when using the serial dilution method, are estimates low?

Back 143

because the artificial and limited growing conditions are not suitable for all organisms.

Front 144

You CFU count for soil bacteria is 88 on a 10*-6 (power) plate. Is the CFU of original gram sample:

A. 88 CFUs
B. 88 x 10*-6 CFUs
C. 8.8 x 10*7 CFUs

Back 144

8.8 x 10*7 (power) of CFUs

Front 145

Define coliforms

Back 145

all aerobic and facultative anaerobic Gneg rods which produce acid and gas from lactose fermentation in 48 hours at 35*C

Front 146

Coliforms

AKA

Back 146

indicator organisms

Front 147

What is the 2nd step of water testing?

Back 147

Confirmed Test

Front 148

What are the three steps of water testing (in order)?

Back 148

Presumptive Test
Confirmed Test
Completed Test

Front 149

What constitutes a positive presumptive test (possible coliforms)?

Back 149

both acid and gas present
(yellow and bubble in durham tube)

Front 150

In a presumptive test what do yellow and blue tubes with no bubbles indicate?

Back 150

Yellow indicates presence of acid and blue means no acid was produced; no bubble means no lactose fermentation.

Front 151

How do you calculate the MPN of a presumptive test?

Back 151

Count the number of positive big tubes; look at chart; count the number of positive baby tubes; look at chart; arrive at answer.

Front 152

Why must the confirmed test be performed for water testing?

Back 152

it is necessary due to the synergism of the mixed bacteria in a tube; giving a false positive.

Front 153

In the confirmed tests (water) name the differential and selective mediums used.

Back 153

Differential medium: EMB Eosin Methylene Blue
Selective medium: EMB Eosin Methylene Blue

Front 154

Describe Eosin Methylene Blue as a differential medium

Back 154

a differential medium because it contains lactose and dyes - it differentiates non-coliforms by dying coliform colonies

Front 155

Describe Eosin Methylene Blue as a selective medium

Back 155

As a selective medium EMB selects Gram negative bacteria

Front 156

why does a green sheen appear on E. coli. EMB agar plates?

Back 156

The acids cause a reduction in pH and precipitate the dye complex (methylene blue eosinate) "dye sink" onto the colonies

Front 157

Identify they areas as indicated.

Back 157

A. E.Coli due to green sheen
B.Enterobacter aerogenes due to pink colonies
C. Non-Coliforms colorless colonies that transmit the reddish-purple color of the medium
D. Enterobacter aerogenes forming purple colonies (note no green sheen)

Front 158

What is a parasite?

Back 158

An organism that lives on or in another organism causing harm.

Front 159

Describe ectoparasites

Back 159

Ectoparasites live on the host
Complex life cycle
multiple morphologies
sexual and asexual reproduction
multiple hosts
protozoa - single celled
motile

Front 160

Describe eukaryotic parasites

Back 160

the human serves as a reservoir of infection

Front 161

What are the major ways that parasites are spread?

Back 161

Via food, water, vectors, soil

Front 162

What is a vector?

Back 162

Organism that transmits an organism from one stage to another
e.g., Anthropods (insects)

Front 163

What is an intermediate host?

Back 163

Intermediate Host provides a living space where parasites asexually reproduce

Front 164

What are some ways that scientists are using to try and stop/slow the spread of parasites.

Back 164

Protective measures for food and water supply
Filtration of water and access to clean water

Front 165

can parasites ever be completely eliminated?

Back 165

Uncontrollable and they can infect multiple hosts that can infect humans

Front 166

Identify the picture.

Back 166

Plasmodium Falciparum (Malaria infection)

Front 167

Identify the picture

Back 167

Ascaris lumbricoides eggs

Front 168

Name the three categories of Helminths.

Back 168

Nematodes - round worms
Cestodes - tapeworms
Trematodes - Flukes

Front 169

Which category contains Ascaris lumbricoides:
Nematodes, Cestodes, Trematodes

Back 169

Nematodes

Front 170

How is malaria vectored?

Back 170

Via the female mosquito Anopheles

Front 171

Name some malaria prevention methods.

Back 171

Prophylactic medications
prevention of mosquito bites
mosquito eradication (insecticides)

Front 172

List some diseases vectored by mosquitos.

Back 172

Malaria
filariases
dengue fever
yellow fever
west nile virus

Front 173

list some diseases vectored by ticks

Back 173

lyme disease
rocky mountain spotted fever
ehrlichiosis

Front 174

If you have a starch agar medium and there is a clear area around the growth line of a bacteria; what test are you looking at?

Back 174

Amylase Test
Positive = clear area around bacteria growth line
iodine reagent used to visualize
exoenzymes present

Front 175

If you have a tributyrin agar medium and there is a clear area around the growth line of a bacteria; what test are you looking at?

Back 175

Lipase
Positive = clear area around bacteria growth line

Front 176

If you have a gelatin agar medium and there is a clear area around the growth line of a bacteria; what test are you looking at?

Back 176

Gelatinase
Positive = clear area around bacteria growth line

Front 177

Of the exoenzyme test what was the trend?

Back 177

Bacillus subtilis tested positive for exoenzymes across all three.

Front 178

Describe Catalase test

Back 178

Tests for the respiration enzyme catalase using substrate hydrogen peroxide
Positive = bubble formation

Front 179

Describe oxidase test

Back 179

tests for the respiration enzyme cytochrome c oxidase
positive = purple spot

Front 180

Describe the nitrate reduction test 1

Back 180

tests for the ability to reduce nitrate to nitrite using nitrate reductase in potassium nitrate broth
positive = bubbles present

Front 181

Describe nitrate reduction test 2

Back 181

tests for nitrite using reagents sulfuric acid and Tromsdorff's reagent
Positive = immediate purple brown

Front 182

Describe nitrate reduction test 2

Back 182

testing for nitrate
zinc added to negative nitrite tests
positive = purple brown
negative = reduction to another gas like ammonia possible

Front 183

For the nitrate reduction tests, what were the two extremes?

Back 183

Bacillus megaterium immediate results;
Pseudomonas aeruginosa N2 gas positive only

Front 184

For the carbohydrate fermentation tests what were the three sugars tested for?

Back 184

Glucose, Lactose, Sucrose

Front 185

Of the carbohydrate tests, what were the two extremes?

Back 185

Alcaligenes faecalis grew on peptones for all three GLS
E.Coli the only one with AG for all three sugars