Microbio Practicum

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biochemical tests can tell 3 things

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1) type of energy used
2) substrates utilized
3) enzymes broken down

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selective media

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suppresses growth of some bacteria but encourages growth of others

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differential media

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distinguishes specific characteristics between organisms
certain species can be identified because they look different

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energy metabolism (2 types)

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respiration and fermentation

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three steps to obtain energy through respiration

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1. glycolysis 2. TCA cycle 3. electron transport chain

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fermentation

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partial breakdown of organic molecules.

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produces of fermentation

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alcohol, acids, hydrogen, carbon dioxide, atp

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key ingredient for respiration

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oxygen

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shape of streptococcus

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gram positive cocci; facultative anaerobe

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requirements of cultivation of streptococcus

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blood agar

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important human pathogens of streptococcus

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s. pyogenes; s. agalactiae; s. pneumoniae

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Hemolysis

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the destruction of red blood cells

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means of detection of hemolysis

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blood agar and candle-in-jar method to reduce oxygen levels. (streptococci are anaerobic organisms)

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beta hemolysis

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complete destruction of red blood cells

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example of beta hemolysis

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streptococcus pyogenes

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alpha hemolysis

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partial destruction of red blood cells

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example of alpha hemolysis

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streptococcus pneumoniae

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Gamma hemolysis

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no destruction of red blood cells

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example of gamma hemolysis

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streptococcus lactis

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shape of staphylococcus

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gram postiive cocci; salt tolerant.

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3 main species of staphylococcus

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staph. aureus
staph. epidermidis
staph. saprophyticus.

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MSA-what it stands for and differential/selective

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mannitol salt agar; selective and differential

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what MSA requires

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mannitol (carbs), 7% NaCl (salt), and pH indicator

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selective factor of MSA

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only bacteria that grows in salty conditions thrive

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differential factor of MSA

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staph. aureus turns from yellow to red because of acid produced

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red bacteria in MSA

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staph. aureus

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shape of enterics

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gram negative, non spore-forming rod shaped

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MacConkey Agar contents

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-bile salt and crystal violet
-neutral red dye
-lactose

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role of bile slat and crystal violet in MacConkey Agar

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inhibits growth of gram-positive bacteria and non-enteric bacteria

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role of neutral red dye in MacConkey Agar

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indicates the pH

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role of lactose in MacConkey Agar

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carbohydrates for lactose fermentation

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selective factor of MacConkey Agar

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only gram-negative will grow from the bile salt and crystal violets

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differential factor of MacConkey Agar

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if enterics form lactose-pink color
if enterics don't form lactose-no change in color

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a pink colored bacteria in MacConkey Agar

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lactose-fermenting enteric present

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TSI test

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Triple Sugar Iron

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contents of TSI

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carbohydrates, peptone, beef extract, yeast extract, sodium thiosulfate, phenol red

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SIM test

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Sulfur reduction, Indole production, Motility

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formation of black precipitate in SIM test

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positive in sulfur reduction

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red after 5 drops of kovac's reagent in SIM test

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positive in indole production

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spread from point of inoculation in SIM test

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positive in motility

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MRVP

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methyl red-vogues proskauer

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methyl red test

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if red after adding methyl red reagent, mixed acid fermentation. none if yellow.

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red in methyl red test

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mixed acid fermentation

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yellow in methyl red test

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no mixed acid fermentation

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vogues proskauer test

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if red after adding vogues proskauer reagent, fermenting bacteria utilize butylene glycopathogen

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red in vogues proskauer test

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fermenting bacteria utilize butylene glycopathogen

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coopper in vogues proskauer test

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fermenting bacteria don't utilize butylene glycopathogen

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Citrate utilization

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determines if bacteria uses citrate as sole carbon source

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contents of citrate utilization

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bromthymol blue dye

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blue in citrate utilization

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high pH

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green in citrate utilization

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low pH

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shape of bacilllus

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gram-positive, spore forming rods

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starch hydrolysis

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whether or not starch is hydrolyzed by bacillus

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enzymes used to hydrolyze starch

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amylase and glucosidase

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contents of starch plate

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beef extract, starch, agar

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blue/black starch plate after iodine addition

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starch hydrolyzed

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MacConkey agar

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uses up oxygen so facultative aerobes can grow; used with blood agar for hemolysis testing.

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mannitol salt agar
yellow-staph. aureus
pink-other staph

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sulfur reduction in SIM

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motility in SIM

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indole production in SIM

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if blue, citrate utilized; if growth, citrate utilized; if neither, not utilized.

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upper-no hydrolyzing enzymes
lower-hydrolyzed

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MRVP (MR)
acid fermentation-red
no acid fermentation-yellow

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MRVP (VP)
butyln glycol pathway used-red
not used-copper/no color

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TSI
yellow-glucose/lactose/sucrose fermentation+acid accumulation (enteric)
red with yellow butt-glucose fermentation with acid production (enteric)
other red slants-no fermentation, not enteric
black -sulfur reduction
cracks-gas production

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kirby bauer antibiotic susceptibility