Lab Practicle

Front 1

Starch Hydrolysis

Back 1

Growth medium used: Starch agar

Reagents used: Flood plate with iodine

Positive result or color: Clear around growth

Negative result or color: Dark blue / brown

Front 2

Lipid Hydrolysis

Back 2

Growth medium used: Tributyrin agar

Reagents used: None

Positive results or color: Clear around growth

Negative results or color: Opaque

Front 3

Casein Hydrolysis (milk protein)

Back 3

Growth medium used: Skim milk agar

Reagents used: None

Positive results or color: Clear around growth

Negative results or color: Opaque (white)

Front 4

Gelatin Hydrolysis

Back 4

Growth medium used: Gelatin

Reagents used: None

Positive results or color: Won't gel

Negative results or color: Solid if cooled in refrigerator

Front 5

Carbohydrate Fermentation (Acid)

Back 5

Growth medium used: Phenol red carbohydrate broth

Reagents used: None

Positive result or color: Yellow

Negative result or color: Red

Front 6

Carbohydrate Fermentation (Gas)

Back 6

Growth medium used: Phenol red carbohydrate broth

Reagents used: (Durham tube)

Positive result or color: Gas bubble

Negative result or color: No gas bubble

Front 7

Oxidase Test

Back 7

Growth medium used: Loopful of nitrient agar.

Reagents used: A few drops of oxidase reagent.

Positive result or color: Purple

Negative result or color: Clear

Front 8

Indole Test

Back 8

Growth medium used: SIM

Reagents used: 10 drops of Kovac's reagent.

Positive result or color: Red

Negative result or color: Yellow

Front 9

Methyl Red Test

Back 9

Growth medium used: Mr-Vp broth

Reagents used: 5 drops of methyl red indicator (set aside aliquot for Voges-proskauer test)

Positive results or color: Red

Negative results or color: Clear

Front 10

Voges-Proskauer Test

Back 10

Growth medium used: Mr-Vp broth (15 min incubation)

Reagents used: Barritts reagents A & B

Positive result or color: Red

Negative result or color: Clear

Front 11

Citrate Utilization Test

Back 11

Growth medium used: Simmon's citrate agar

Reagents used: None

Positive result or color: Purssian blue

Negative result or color: Green

Front 12

Hydrogen Sulfide Production

Back 12

Growth medium used: SIM or TSI

Reagents used: None

Positive results or color: Black

Negative results or color: Clear

Front 13

Motility Test

Back 13

Growth medium used: SIM

Reagents used: None

Positive results or color: Cloudy throughout

Negative results or color: Cloudy only at top

Front 14

Urease Test

Back 14

Growth medium used: Urease broth

Reagents used: None

Positive results or color: Hot pink

Negative results or color: Light yellow

Front 15

Phenylalanine Deamination Test

Back 15

Growth medium used: Phenylalaine agar

Reagents used: A few drops of 10% ferric chloride

Positive results or color: Green at top

Negative results or color: Agar stays clear

Front 16

Nitrate Reductase Test

Back 16

Growth medium used: Nitrate broth

Reagents used: 5 drops of A & B / Minute quanity of C

Positive results or color: Red (after A & B)

Negative results or color: Red (after reagent C)

Front 17

Given a plate of MSA, upon which a pure culture of bacteria has been allowed to incubate for 24 hours, determine: Whether the culture could be staphylococcus or not and explain why or why not.

Back 17

*Staphylococcus*

Because mannitol salt agar ferments gram positive staphylococcus bacteria. (Salt Loving)

Front 18

Given a plate of MSA, upon which a pure culture of bacteria has been allowed to incubate for 24 hours, determine: Which species (approximately) of Staphylococcus is represented, if it is a staph.

Back 18

S. Aureus - Positive

S. Epi - Negative

S. Sapro - Negative

Front 19

Given a plate of MSA, upon which a pure culture of bacteria has been allowed to incubate for 24 hours, determine: The full name of the (MSA) medium and describe it's primary ingredients.

Back 19

(Mannitol salt agar) this medium contains a high salt concentration 7.5% NaCl which is inhibitory to the growth of most, but not all, bacteria other than the stapylococci. Enriches for gram positive! The indicator on the plate is phenol red.

Front 20

Recognize and explain how the DNAse agar is used to reconfirm the identification of Staphylococcus Epidermidis and Staphylococcus Saprophyticus and describe how they differ.

Back 20

Coagulase-positive staphylococci also produce the hydrolytic enzyme DNase; thus this test can be used to reconfirm the identification of S. aureus.

-S.aureus is positive has white surrounding/uses the DNAse.

-S. sapro, S.epi are negative with blue surrounding.

Front 21

MSA

Back 21

Yellow halo surrounding their growth = fermenting Staph will grow on the presence of catalase while the enteric will not.

Front 22

DNAse AGar

Back 22

-A rose pink halo around the area the area of growth

-Absence of halo is indicative of a negative result and the inability of the organism to produce DNAse.

*OUR class had different agar! WHITE was POSITIVE. S. aureus.

Front 23

Identify the primary laboratory tests used to differentiate Staphylococcus Epidermis and Staphylococcus Saprophyticus and describe how they differ.

Back 23

Novobiocin Test

Front 24

Novobiocin Test

Back 24

Is used to differentiate between the 3 staphs, the Mueller-Hinton agar plate is heavily seeded with the test organism to produce a confluent growth on the agar surface. (Kirby Bauer method is used.)

-S. saprophyticus is the only resistant one

-S. Aureus and S. Epidermidis are sensitive

Front 25

Coagulase Test

Back 25

-Clot formation indicates a positive result

-The absence of coagulation is a negative result

Production of coagulase is indicative of an S. aureus strain. The enzyme acts within host tissues to convert fibrinogen to fibrin.

-Clot formation within 4 hours is interpreted as a positive result and indicative of a virulent S. aureus strain the absence of coagulation after 24 hours of incubation is a negative result, indicative of an avirulent strain.

Front 26

List the three genera of pyogenic cocci

Back 26

1. Gonococci

2. Streptococci

3. Staphylococci

Front 27

Streptococcus pyogenes

Back 27

Beta Hemolytic

Front 28

Streptococcus pneumoniae

Back 28

Alpha hemolytic

Front 29

Staphylococcus aureus

Back 29

Beta hemolytic

Front 30

Staphyococcus epidermidis

Back 30

Gamma hemolytic

Front 31

Identify (name) a quick & simple bio-chemical test to differentiate Streptococcus and Staphylococcus species.

Back 31

Catalase Test. If it bubbles it's staphylococcus positive

Front 32

Identify (name) the small inverted tube used in the carbohydrate fermentation test state its function and how to interpret the results shows.

Back 32

Durham tube

Front 33

Identify at least one major clinical test used to differentiate Streptococcus pneumoniae from other alpha-hemolytic streptococci.

Back 33

Optochin Sensitivity test.

Front 34

Describe the use and application of the optochin ("p") or ("o") disk with streptococci and, provided with the name or a sample of optochin disks, identify the bacterial species that is used to differentiate from other streptococci.

Back 34

Used to differentiate Streptococcus pneumonia from other streps. S. pneumonia shows zone's of inhibition (sensitive/dying)

Front 35

Explain and recognize how to inoculate and interpret the results of TSI agar.

Back 35

Triple Sugar Iron Agar! Is designed to differentiate among the different groups or genera of the Enterobacteriaceae, which are all gram-negative bacilli capable of fermenting glucose with the production of acid, and to distinguish the Enterobacteria from other gram-negative intestinal bacilli.The slant is inoculated by means of stab-and-streak procedure.

Front 36

List at least four different genera of bacteria which are considered to be "Enteric bacilli" (Enterobacteriacaea) and describe at least two important characteristics they all share.

Back 36

Citrobacter

Enterobacter

Escherichia

Klebsiella

Proteus

Pseudomonas

Front 37

Identify (name) each of the four bacteriological biochemical tests represented by the mneoric IMViC, and describe the significant of this battery of tests.

Back 37

Indole, Methyl red, Voges-Proskauer, Citrate Utilization

Front 38

NA

Back 38

Nutrient Agar

Front 39

BA or BAP

Back 39

Blood agar or Blood agar plate

Front 40

NB

Back 40

Nutrient Broth

Front 41

MR-VP

Back 41

Methyl Red-Voges Proskauer

Front 42

EMB

Back 42

Eosin Methylene Blue

Front 43

SIM

Back 43

Sulfide Indole Motility

Front 44

Identify the primary laboratory tests used to differentiate Staphylococcus epidermidis and Staphylococcus saprophyticus and describe how they differ.

Back 44

Novobiocin Test- Differentiates between the three staphs.

1. S. epi- Sensiitive

2. S. aureus- Sensitive

3. S. sapro- Resistant

Front 45

Identify the Lancefield's group that is associated with scarlet fever and strep throat.

Back 45

Group A Strep

Front 46

Identify (name) describe and interpret the results of the standard test used to determine if a beta hemolytic streptococcus belongs to Lancefield's group A. # 58 pg. 159

Back 46

Bacitracin Sensitivity Test (A disk)

Front 47

Identify the specific group of streptococci that are identified through the use of the bacitracin ("A") disk.

Back 47

Group A strep / Streptococcus pyogenes

Front 48

Identify (name) describe and interpret the results of the standard test used to determine if a beta-hemolytic streptococcus belongs to Lancefield's group A.

Back 48

Bacitracin Sensitivity Test. Group A strep / Streptococcus pyogenes