GRAM STAIN

- purpose of stain
- step 1-4 and purpose 

- allows for colorless bacteria to be visualized in clinical specimens and in culture media

1. CV- adheres to pepgly cell wall

2. Iodine inc. affinity for the dye 

3. Ethanol decolorizes

4. Safranin counterstains 

Kinyoun acidfast

- specific for ____

- describe how af bacteria react

- Red results

- blue results

-Specific for  mycobacteria

- af is the resistance of bacteria to decolorize via acids during staining.

-RED: non-acid (-)

-BLUE: acid-fast (+)

Auramine-Rhodamine stain

- aka the ____ stain

- histologically visualize ___ bacteria with flourescence microscopy
- these bacteria display ___ flourescence 

- aka the Traunt auramine-rhodamine stain

-visualize acid-fast ( Mycobacterium)

-display reddish-yellow flourescence

Hemolysis with blood agar

- alpha colonies

-beta colonies

-gamma colonies

- brown and green erythrocytes

- erythrocytes surrounded by a clear zone

- no growth

Catalase Test:

- what is catalase

- use this substance on bacteria

- what indicates a positive test

- enzyme in STAPHLOcocci and MICROcocci but not STREPTOcocci

- 3% H2O2

- bubbles

Coagulase Test:

- what is coagulase and what does it do

- what indicates a positive test

- what species would be (+) and which (-)

-enzyme produced by STAPHYLOcocci. Used as a defense mechanism a clots plasma

-white clumps

-Staph aureus will be (+)

- Staph epidermidis will be (-)

Optochin

- specific for what bacteria

-what type of agar is used

-what type of zone indicates susceptibility

- for strep. pneumococcus

- blood agar

- Zone of inhibition greater than 14mm

Bacitracin test:

- what bacteria is this test specific for

- what is bacitracin

-what type of agar is used

- distinguishes Strep. pyrogenes from other

- an antibiotic that impairs cell wall syn. of some gram+ bacteria (i.e s.pyrogenes)

-blood agar and noting any zone of inhibition

Novobiocin test:

-what does novobiocin do

-used to differentiate what two bacteria

- what indicates susceptibility

- inhibits bacterial DNA gyrase

-staph SAPROPHYTICUS from other coagulase(-) staph

- zone of inhibition greater thatn 12mm

Nitrocefin Test:

-indentifies what type of bacteria

- RED results around disk

- YELLOW results around disk

- beta-lactamase producing bacter

- RED or (+) produces beta-lactamase

- YELLOW or (-) no blac

Oxidase Test:

-identifies what genus of bacteria

PYR test: 

-used to identify what stype of species

-determines that lack of presence of what

- what indicates a positive test

-Enterococcua species

- pyrolidonyl amino peptidase

- PINK/RED around PYR disk

Indole test:

-detects the product of what

- differentiates what species of bacteria

- tested on oxidase__, lactose __  and gram __ bacteria

- BLUE means

- PINK means

- tryptophanase

- differentiates Enterobacteriaceae

- negative, positive, negative rod

- positive

-negative

Bile esculin test:

- distinguishes between what two cocci

-nutrient agar made of ___ and ___

- Black medium means

- unchanged medium means

- Streptococci and enterococci

- agar made of bile and esculin

- Enterococci faecalis and resistant to agar

- Strepococci pyrogenes cannot hydrolize esculin

NaCl Test:

-Distinguished what two bacteria

- what agar is used

- if colonies grow it is

- if colonies do not grow it is

- streptococci and enterococci

- chocalate agar

- Enterococci

-Streptocicci

CAMP test:

-distinguish _____ from other streptococci

because this group produces ___

-streak plate vertically with ___  and streak plate perpendicular with GroupB strep.

- what indicates a positive

-GroupB, CAMP factor

- staphylococcus aureus

-arrow head means GroupB strep is present

Motility Test

- used semisolid agar and inocualte via __

- if it is an obligate aerobe it will __

- specific for what bacteria and shape it takes in the agar

- stabbing

- all migrate to the top of the agar creating a "buldge" - umbrella

- Listeria moncytognes 

Sheep blood agar:

- made of __ and 5% __

- supports growth of __

- nutrients , blood

-pathogens

MacConkey Agar
-inhibits growth of ___

- what does red/pink signify

-clear colonies means

- gram positive bacteria

-lactose fermentation

- no lactose fermentation

XLD agar:

- selective for presence of __,__,__,__

-Salmonella will apear

-Shigella will appear

- Enterobacter will appear

-xylose, lactoase, surcrose and lysine

- red w/ black center do to pH and H2S

-red or colorless

-yellow do to use of carbs

Choclate agar:

- how does is differ from blood agar

- why is is heated

- allows what  two bacteria to grow

- it is heated to 80C for 15 min

- release hemoglobin and growth factors

- N. gonorrhaeae and H. influenza

Mannitol salt agar:

- high content of __ to inhibit growth of organisms found on ___

- differentiates ___ due to pH indicator

- NaCl , skin

- mannitol fermenting staphylococci 

Mueller-Hinton agar: 

- agar contains ___ and ___

- what species of bacteria grow well on this agar

- test antibiotiv susep. for __ and __

- starch and casamino acids.
- Neisseria grow well on this medium

- staph and enteric gram neg. rods 

Thayer- MArtin agar:

- selective for  isolation of ___ and ___ from material containing normal flora

Sabourand dextrose agar:

-used to isolate ___

-what inhibits bacterial growth

- pathogenic and nonpathogenic fungi

-  acidic pH  inhibitsbacterial growth

Enriched media:

-supports the growth of 

-example of enriched media

-wide variety includding fastidious ones

-ex is blood agar

selective media:

-prevent the growth of ___ to select for __

Nutritie media

- non ____

oxidation reaction
-can only metabolize ___ under__

- use media that contains __ and ___

- one tube is open and one tube sealed w/ oil to make___

-oxidizers in what tube and what color

-fermenters in what tube

- carbs uner aerobic conditions

- glucose, pH indicator

-anerobic conditions

-oxi. produce YELLOW in open tube only

-fer. will be in both open and closed tubes

Fermentation reaction: IMPORTANT
-test the ability to ferment __ or __ and divides __ bacteria from __ bateria 

- use a test tube w/ __ and __

- pH indicated if __ was formed through fermentation

-lactose, surcrose, pathogenic, nonpathogenic

-sugar, pH indication

-acid

API biochemical strip:

- specfic what what species of bacteria

- how many wells

-Enterobacteriaceae and gram-

-23 wells

Agar disk diffusion test:

Agar gradient diffusion test ( Etest)

- determines the least amount of  ___

- Etest strips are  are coated with

- an elliptical zone will be formed, where will it be "read"

- antimicrobial agent that inhibits growth of a microragaism

- certain concentration gradients of antimic

-read where  bacteria intersect at the strip

Agar dilution test

- ___ of antibiotic in agar to see the __________

- serial dilution , least amount that inhibits growth

Microbroth Dilution test:

- automated to test ___

Minimum Inhibitory concentration:

-test organism for 

- what is the MIC

-susceptibility to antibiotics 

- the lowest concentration of animicrobial agrent that inhibis gorwth of a bacterium

Satellitism

-distinguished what species of bacteria
- What three disc will be used

- zone around X- only means

-zone around V-means

-zone around XV disc

-Haemophilis

- exogenous hemin X, NAD V or XV disc

- requirement for heme

-requirement for NAD
- need for both like H. influenae 

Sterility test w/ Bioloical indicator:

- importan for what type of instruments

- what is used to standardize or compare

-PURPLE means

- YELLOW means

-non autoclavable

- suspended spores of Bacillus staeothemophilus

-Sterile and spores are inactive

- imcomplete sterilization and spores have growth

Complement fixation:

 - used to detect is there is