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237 Cards in this Set
- Front
- Back
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what is the first thing and last thing you must do to your lab bench workspace when you workin in the micro lab
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disinfect it/sanatize it with disinfectent HDQ
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the total magnification while using the "scanning" objective lense with a 10x ocular lens is __ and with a "high-dry" objective is __
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40x, 400x
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what are two ways that you could improve the resolution of a microsope?
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-decrease wavelength
-increase numerical aperture |
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it is always necessary to add __ to use the 100x objective lens
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oil
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1 meter is equal to __ microns
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1000000
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1 micron equals __ nanometers
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1000
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considered non-living, usually the smallest microbe
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virus
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eukayotic, single-celled, usually motile
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protozoa
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eukayotic, usually photosynthetic, can be multicellular
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algae
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prokayotic
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bacteria
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preparation of bacterial smears must be done without introducing unwanted microbes or contaminants into pure cultures. this is accomplished by using __
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aseptic technique
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__ staining is a good way to stain bacteria that don't stain well with other types of stains
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negative stain
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__ staining provides a very accurate view of the actual size and shape of bacteria
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negative staining
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what is one bacterial genus for which negative staining would likely be used
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treponema palladin
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the gram stain is a __ stain
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differential
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the best reason to perfrom a gram stain on bacteria isolated from teh urine a patient with a bladder infection is which __ might be most effective
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antibiotic
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suppose you perform a gram stain of what you think is a pure bacterial culture and you see a mixture of pink cocci and purple rods. the best conclusion is __
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the culture is contaminated
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suppose you perform a gram stain of what you think is pure bacterial culture and you see a mixture of pink and purple cocci. the best conclusion is __
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the culture is old
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partially digested proteins in bacterial media are called __
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peptones
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the minimum temperature required for autoclaving is __ degrees celcius for __ min at __ psi
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121, 15, 15
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after autoclaving, agar becomes a solid at after it cools to about what temperature
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45 degrees celcius
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agar is a __ derived from teh cell walls of an __
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plysacchride, alga
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the acid fast staining procedure is usually performed to detect bacteria of teh genus __, which can cause the disease __
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mycobacterium, TB
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acid fast negative bacteria stain what color?
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blue
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the primary stain used to detect endospores is called __
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malachite green
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when looking at a bacterial smear stained for endospore, teh pink-staining cells are the __
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vegetative cells
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the two geni of endospore-forming bacteria are __
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clostridium and baccillus
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EMB agar is both __ and __
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differential and selective
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__ is selective because it contains a high amount of sodium chloride
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mannitol salt agar
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a __ medium favors the growth of certain types of bacteria
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enrichment
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starch is an example of a __
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carbohydrate (polysacchride)
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the exoenzyme that digests starch is called __
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amylase
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in order to read the results of a starch agar plate you must add which reagent?
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iodine
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in a positive reaction on starch agar, what would appear around the inoculated bacteria
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a zone of clearing, surrounded by dark purple
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if you inoculated a TSI tube with a pure bacterial culture and incubated overnight, what could you conclude if only the butt turned yellow?
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only 1 sugar was fermented
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what is the substrate of teh enzye beta-galactosidase
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lactose
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before interpreting results of metabolic tests, how can you determine whether bacterial growth occurred in the medium?
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turbidity
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the methyl red test detects the products of which fermentation pathway?
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mixed acid
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forms indole
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trptophanase
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cuases the pH of teh medium to increase
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uresase
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enzyme that produces H2S
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cysteine desulfhydrase
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enzyme required for positive gelatin hydrolysis test
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collagenase
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suppose you inoculate a tube of gelatin medium, incubate overnight, and teh next day you check the tube in the 37 degree incubator and it is a liquid. what is the best explanation?
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cannot interpret because it has not cooled yet
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for the clinical test mentioned lab, in which patients drink radioactive urea, what is actually being measured in the patients breath?
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redioactive CO2
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the breath test in which a person drinks radioactive urea is used to help diagnose which bacterium?
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H. pylori
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kovac's reagnet is used to detect what specific molecule?
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indole
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in a SIM test, H2S production is indicated by __
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a black precipitate
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all broken glass/ glass to be disposed goes in teh __
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broken glass box
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petri dishes, swabs, etc go in teh __
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autoclave box
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save/recycle __
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innoculation tubes
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the ocular lens has a magnification of __
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10x
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total magnification=
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ocular mag x objective mag
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the 4x objective is called the __ objective
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scannig
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the 10x objective is called the __ objective
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low
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the 40x objective is called the __ objective
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high, dry
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the 100x objective is called the __ objective
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oil
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resolution = __
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wavelength/2xnumerical aperture
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you want a __ resolution
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small
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how does a blue filter give you smaller resolution?
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it has a smaller wavelength of light
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how does oil give a smaller resolution?
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oil has the same refraction index as glass, the oil keeps teh light from bending
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means that teh 4x, 10x, 40x, and 100x are all in focus
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parfocal
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what is the main difference between eucaryotes and prokaryotes?
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organelles
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protozoans are typically __ and __
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motile, heterotrophs
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algae are typically __ and __
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nonmotile, autotrophs
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the two branches of procayotes
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eubacteria (true bacteria) and archeaa
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the average size of a bacterium is __
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1-10 microns
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the average size of a protist is __
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10-5 microns
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the average size of an RBC is __
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7-10 microns
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the average size of a virus is __
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nanometers
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resolution (d) of a light microscope is __
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.2 microns
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the resolution of teh human eye is __
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200 microns
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true/false: aseptic = sterile?
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FASLE!!
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working with a desired microbe but avoiding contamination, working with pure cultures
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aseptic
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3 type of sprial bacteria
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-virbro (bean)
-spirillum (squiggle) -spirochete (curly q) |
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__ stain does not alter the shape, or size of bacteria at all
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negative
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what kind of bacteria shape would you probably perform a negaive stain on?
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spirochetes
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what bacteria causes lyme disease?
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borrella burgdorferi
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what is the first step of gram staining?
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smear, dry, heatfix
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how long do you apply the primary stain in a gram stain?
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30 sec
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how long do you apply the mordent stain in a gram stain?
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1 min
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how long do you add the decolorizer
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to effect, 10-30 seconds
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how long do you add the counter stain?
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1-2 min
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in the gram stain mechanism, the __ stain enters the cytoplasm of gram positive and gram negative bacteria
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primary stain (crystal violet)
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in the gram stain mechanism, the __ stain forms crystals too large to escape gram + cell wall
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mordent stain (grams iodine)
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in the gram stain mechanism, the __ dissolves the outer membrane of gram -
-CV/iodine complex can escape through thin PG layer |
decolorizer (alcohol)
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in the gram stain mechanism, the __ stain counterstains gram - red.
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counter stain (safronin)
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some gram + become gram - as the culture __
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ages
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a media in which the exact chemicals are defined
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chemically defined media
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a media in which you do not know the exact ingredients.
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complex media
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a pancreatic enzyme used to digest ttyptic soy peptone
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trypsin
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TSA=
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tryptic soy agar
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TSB=
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tryptic soy broth
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most bacterium don't eat __, which is why it is used in solid medias
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agar
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agar melts at ~__ degrees celcius and stays a liquid until __ degrees celcius
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100, 45-50
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no living microbes
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sterile
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steam sterilization
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autoclaving
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to put bacteria into something
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innoculate
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when bacteria cloud the tube after innoculation indicating growth
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turbidity
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used for detection of bacteria with mycolic acid in their cell wall
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acid-fast staining
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bacteria that stain acid fast positive are __
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red
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bacteria that stain acid-fast negative are __
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blue
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type of bacteria that stain acid-fast positive
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mycobacterium
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the process of making endospores
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sporulation
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what kind of bacteria create endospores?
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gram +
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what is the first step in an endospore stain?
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smear, dry, heatfix
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under what conditions and how long should you apply the primary stain (malachite green) for an endospore stain?
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over heat for 3 min
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what are the steps for an endospore stain?
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-smear dry heatfix
-primary stain (malachite green) 3 min over heat -decolorize with water -counterstain with safronin |
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what do you decolorize an endospore stain with?
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water
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what should you do in between each steak on a steak plate?
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flame and cool the loop
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what are three ways to ID bacteria?
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-biochemical (metabolic)
-immunological (antibodies) -genetic (PCR) |
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a media in which some bacteria grow and some don't
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selective
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a media that favors/encourages certain bacteria
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enrichment
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a media that gives additional information about organisms that grow on the medium
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differential
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mannitol salt agar is a __ and __ media
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selective and differential
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what makes mannitol salt agar selective and what does it select for?
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high NaCl, selects for gram +
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what makes mannitol salt differential and what does it tell us?
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it tells us there is acid production, mannitol is the only sugar that some bacteria will ferment
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how can we tell that a bacteria ferments mannitol on a mannitol salt agar plate?
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turns solution yellow when a pH indicator is added (phenol red)
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what is a type of bacteria that ferments mannitol?
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staphylococcus aureus
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EMB (eosin methonal blue) is a __ and __ media
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selective and differential
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EMB is selective for __
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gram - bacteria
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what makes EMB differential?
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lactose (glucose+galactose) (must be able to split to ferment)
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if a bacteria ferments on an EMB plate it will turn __ meaning it produces __
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green, beta-galactosidase
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lactose and sucrose are examples of __
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disaccharides
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when grams iodine is added to a starch agar plate, the plate forms a dark purple color. if there is a clear ring around a line of bacteria, it indicates __
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the absence of starch (amylase is produced)
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what is incorporated into fermentation tubes to test for the presence gas
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durham tube
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a fermentation tube contains water, salts, peptones, phenol red, and __ sugar(s)
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1
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TSI (triple sugar iron agar) contains __ sugars
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3 (glucose, sucrose, lactose)
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what are the concentrations of sugars in a TSI tube?
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glucose: 1x
sucrose: 10x lactose: 10x |
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if the but of a TSI tube is yellow that indicates __
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glucose only fermented
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if the slant but of a TSI tube is yellow that indicates __
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glucose + sucrose and or galactose is fermented
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what makes EMB differential?
|
lactose (glucose+galactose) (must be able to split to ferment)
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if a bacteria ferments on an EMB plate it will turn __ meaning it produces __
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green, beta-galactosidase
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lactose and sucrose are examples of __
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disaccharides
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when grams iodine is added to a starch agar plate, the plate forms a dark purple color. if there is a clear ring around a line of bacteria, it indicates __
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the absence of starch (amylase is produced)
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what is incorporated into fermentation tubes to test for the presence gas
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durham tube
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a fermentation tube contains water, salts, peptones, phenol red, and __ sugar(s)
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1
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TSI (triple sugar iron agar) contains __ sugars
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3 (glucose, sucrose, lactose)
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what are the concentrations of sugars in a TSI tube?
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glucose: 1x
sucrose: 10x lactose: 10x |
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if the but of a TSI tube is yellow that indicates __
|
glucose only fermented
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if the slant but of a TSI tube is yellow that indicates __
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glucose + sucrose and or galactose is fermented
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a media in which some bacteria grow and some don't
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selective
|
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a media that favors/encourages certain bacteria
|
enrichment
|
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a media that gives additional information about organisms that grow on the medium
|
differential
|
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mannitol salt agar is a __ and __ media
|
selective and differential
|
|
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what makes mannitol salt agar selective and what does it select for?
|
high NaCl, selects for gram +
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|
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what makes mannitol salt differential and what does it tell us?
|
it tells us there is acid production, mannitol is the only sugar that some bacteria will ferment
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how can we tell that a bacteria ferments mannitol on a mannitol salt agar plate?
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turns solution yellow when a pH indicator is added (phenol red)
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what is a type of bacteria that ferments mannitol?
|
staphylococcus aureus
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EMB (eosin methonal blue) is a __ and __ media
|
selective and differential
|
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EMB is selective for __
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gram - bacteria
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bacteria that stain acid-fast negative are __
|
blue
|
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type of bacteria that stain acid-fast positive
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mycobacterium
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the process of making endospores
|
sporulation
|
|
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what kind of bacteria create endospores?
|
gram +
|
|
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what is the first step in an endospore stain?
|
smear, dry, heatfix
|
|
|
under what conditions and how long should you apply the primary stain (malachite green) for an endospore stain?
|
over heat for 3 min
|
|
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what are the steps for an endospore stain?
|
-smear dry heatfix
-primary stain (malachite green) 3 min over heat -decolorize with water -counterstain with safronin |
|
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what do you decolorize an endospore stain with?
|
water
|
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what should you do in between each steak on a steak plate?
|
flame and cool the loop
|
|
|
what are three ways to ID bacteria?
|
-biochemical (metabolic)
-immunological (antibodies) -genetic (PCR) |
|
|
media that in which some bacteria grow and some dont
|
selective
|
|
|
media that favors/encourages certain bacteria to grwo
|
enrichment
|
|
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media that gives additonal infor aobut oraisms that grow on the medium
|
differential
|
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mannitol salt agar is a __ and __ media
|
differential and selective
|
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how is mannitol salt agar selective and what does it select for?
|
high NaCl, gram +
|
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how is mannitol salt agar differential and what does it test for
|
mannitol is the only sugar taht some bacteria will ferment, acid production
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if a bacteria ferments mannitol, it turns __ with a pH indicator (phenol red)
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yellow
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__ is a bacteria that will ferment menitol
|
staphylococus aureus
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EMB is a selective medium for __
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gram -
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EMB is differnetial because it conains __
|
lactose
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if an EMB test is +, bacteria will ferment and cause the media to turn __. this shows the presence of __
|
green, beta-galactosidase
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lactose and sucrose are examples of __
|
disacchrides
|
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when grams iodine is added to a starch agar plate, a clear ring around the bacteria line indicates __
|
the absence of starch (amylase is produced)
|
|
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fermentation tubes include a __ to see if gas is produced
|
durhams tube
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fermentation tubes include water, salts, peptones, phenol red, and __ sugar(s)
|
1
|
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TSI (triple salt agar) contains __ sugar(s)
|
3
|
|
|
what are the sugar concentrations in a TSI tube?
|
glucose: 1x
sucrose: 10x lactose: 10x |
|
|
if the but of a TSI tube is yellow, that indicates __
|
only glucose was fermented
|
|
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if the slant and but of a TSI tube is yellow, that indicates __
|
glucose + sucrose and or lactose was fermented
|
|
|
MRVP (methyl-red, voges-prostauer) test test for the fermentation of __ and the production of __
|
mixed acid, acetoin
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the MR part of an MRVP test will be + if teh pH is __ and will turn __
|
<4, yellow
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if the VP test of an MRVP test is +, it produces __ and the medium turns __. if it is negative, the medium turns __
|
acetoin, yellow, light brown
|
|
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the citrate test tests for the presence of __
|
citrate permease
|
|
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what kind of bacteria produces acetoin?
|
enterobacter aerogenes
|
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if a citrate test is + the pH will __ because __ is being produced.
|
increase, CO2
|
|
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if a citrate test is + the medium will go from __ to __
|
green, blue
|
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__ is derived from proteins found in connective tissue ie collagen , elastin
|
gelatin
|
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some bacteria produce exoenzymes (gelainase and collagenase) to break down proteins. this helps the bacteria spread in tissues of live hosts. this is called a __
|
virulence factor
|
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in a nutrient gelatin test, you incubate the innoculated tube at __ degrees celcius overnight and then bring back to room temp to test
|
37
|
|
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after being brought to room temp, a nutrient gelatin test is + if the media is a __ and - if the media is a __
|
liquid, solid
|
|
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__ is produced by animals as nitrogenous waste product
|
urea
|
|
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an enzyme that breaks done urea
|
urease
|
|
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if a bacteria produces urease, the pH of a medium will __
|
increase
|
|
|
if a urea agar test is positive, the color will go from __ to __
|
yellow, fushia
|
|
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what bacteria produce urease and would therefore be + on a urea agar test?
|
Helicobacter Pylori
|
|
|
urease is a __ because it increases the pH of the stomach in order to make it more hospitible
|
virulence factor
|
|
|
enzyme that uses NH3
|
deaminase
|
|
|
enzyme that uses COOH
|
decarboxycase
|
|
|
enzyme that uses SO4
|
desulfanase
|
|
|
what is the test for H2S called?
|
SIM
|
|
|
a SIM test can measure __, __, and __
|
H2S, indole, motility
|
|
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what causes the H2S to precipitate is a SIM test?
|
iron
|
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if bacteria in a SIM are motile, they will __
|
spread out from the stab line
|
|
|
f bacteria in a SIM produce H2S, there will be a __
|
black precipitate
|
|
|
if a bacteria is + for H2S in a SIM test, that bacteria can break down __
|
cystein
|
|
|
in order to test for indole on a SIM test, you must add __. if +, it will turn __
|
kovacs reagent, red
|
|
|
what does it mean if bacteria are positive for indole in a SIM test?
|
they produce tryptophanase
|
|
|
how do you do a catalase test on bacteria?
|
put H2O2 on them.
|
|
|
if a bacteria is catalase +, it will __ when H2O2 is placed on it.
|
bubble
|
|
|
the point of __ is to clean up products of respiration, and can encourage spread in tissue
|
catalase
|
|
|
the production of catalase can help bacteria survive against __ by counterbalancing the respiratory burst
|
macrophages
|
|
|
an oxidase test is used to determine whether or not a bacteria produce __
|
cytocrome C oxidase
|
|
|
the final carrier in the electron transport chain
|
cytocrome C
|
|
|
if a bacteria is + in an oxidase test, it forms a __ substance
|
dark purple
|
|
|
uses inorganic electron receptor at the end of the ETC (ie nitrate)
|
anaerobic respiration
|
|
|
if "A" and "B" are added to a nitrate reductino test tube and it is +, it will turn
|
red
|
|
|
if no color change occurs in a nitrate reductino test tube after "A" and "B" are added, it is __
|
negative
|
|
|
if no color change occurs in a nitrate reductino test tube after "A" and "B" are added, it is negative. is this definitive?
|
NO!!!
|
|
|
once zinc is added to a a nitrate reductino test tube after "A" and "B" are added and the tube turns pink, it is __ for nitrate reduction
|
negative
|
|
|
once zinc is added to a nitrate reductino test tube after "A" and "B" are added and there is no color change, it is a __ for nitrate reduction
|
positive
|
|
|
__ is an oxygen growth test media that is a reducing medium
|
sodium thyoglycolate
|
|
|
pasteurizaton __ microns
|
decreases
|
|
|
pasteurizaton can either be at __ degrees celcius for 30 min, or at __ decrees celcius for 15 seconds
|
63, 72
|
|
|
a rare event involving the aquisition by a recipient bacterium of small pieces of DNA released from a dead donor bacterium
|
transformation
|
|
|
when transcription is activated by the presence of a particular substrate, the gene has an __
|
inducible operon
|
|
|
when the promotor is always turned on
|
constituative
|
|
|
teh gene with the constituitive promotor in teh pGLO plasmid is __
|
AMP
|
|
|
teh gene with the inducible operon in teh pGLO plasmid is __
|
GFP
|
|
|
GFP in teh pGLO plasmid gene is from __ and cuases the pGLO to glow under __
|
jellyfish, UV light
|
|
|
GFP is an inducible operon in pGLO when __ is present
|
arabinose (ABA)
|
|
|
AMP in the pGLO plasmid codes for an enzyme that alters/inactivates __
|
ampicillin
|
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|
what were 2 things done to make ecoli competent (willing to take up DNA)?
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-CaCl2
-heatshock |
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what kind of e.coli were used in the pGLO experiment?
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E. coli HB101
|
|
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resident microbia on your hands are __
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always present
|
|
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transient microbia on your hands are __
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picked up
|
|
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populations or comunities of microorganisms that attach and grow on a solid surface that has been exposed to water
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biofilms
|
|
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the ability of bacteria to communicate and coordinate behavior
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quarum sensing
|
