Unknown ID Charts

Unknown ID Charts The following charts give the EXPECTED assay outcomes for each Genus and species of bacteria used as unknowns in your lab. As with all living things, there are numerous strain differences within a given species, and a particular culture may not give the expected result. Blank spaces within the chart mean that data does not exist for that test, or that the test is not diagnostic for that Family/species. Key to symbols: + = positive result for >70% of strains - = negative result for >70% of strains V= variable, strain dependent trait, cultures may give + or - result A = Acid AG = Acid and gas a = obligate aerobe fa = facultative anaerobe b = bacillus c = coccus Key to All Tests Gram Stain – reaction in Gram stain, (+) = retains crystal violet; (-) = stains pink from safranin; results can be confirmed using PEA, MAC and KOH Shape – cell shape – (c) coccus, (b) bacillus Endospore – results of endospore stain; (+) = endospores present as greenish blue bodies on slide, non- spore structures are pink; (-) = no blue-green bodies present, only pink present, no endospores Salt Tolerance – based on results of MSA plate. MSA inhibits the growth of organisms that cannot tolerate ≈7.5% NaCl; (+)= growth, organism is a halophile; (-) = no growth, no salt tolerance, organism is not a halophile KOH – results of KOH test; (+) Gram-negative organisms will produce a sticky, viscous lysate, (-) no viscous lysate, organism is Gram-positive PEA – phenylethyl alcohol agar – presence of 2.5% phenylethyl alcohol inhibits gram-negative bacteria - giving poor, if any, growth; (+) = growth, Gram-positive; (-) = no growth, Gram-negative MAC – MacConkey Agar – inhibits the growth of Gram+ organisms due to presence of crystal violet; (+) = growth, Gram-negative; (-) = no growth, Gram-positive SIM (Motility) – results of SIM medium; (+) = motile, organisms will migrate outwards from original stab; the medium will turn hazy beyond the original stab; be sure to compare to an uninoculated tube as a negative control. (-) no growth beyond stab, non-motile Thioglycollate Broth– oxygen requirements - results of growth in sodium thioglycollate medium; obligate aerobes (a) will grow only on surface, in area originally defined by pink resazurin band; facultative anaerobes (fa) will grow throughout medium, with heaviest growth on surface Carbohydrate Utilization – results of fermentation (glucose, lactose, sucrose, mannitol); (-) is characterized by red, pink or orange phenol red, no fermentation; acid production (A) is indicated by change in phenol red from red to yellow; acid and gas production (AG) is characterized by yellow color and presence of bubble in Durham tube. Simmons Citrate – citrate utilization - results of Simmons citrate slant; this is a selective media, only those organisms that can utilize citrate can grow; (+) = a change in color from green to blue, or obvious abundant growth; (-) = no citrate utilization, green or no growth Nutrient gelatin - gelatin hydrolysis – (+), gelatin will be liquid at room temperature, or form liquid crater; (-) = gelatin will remain solid at room temperature Starch Plate - starch hydrolysis, (+)a zone of clearing around the bacterial growth, remainder of plate will stain dark blue to purple/black due to interaction of starch and iodine; (-)= no zone of clearing will be present around growth Oxidase – results of oxidase dry slide, detects presence of enzyme cytochrome oxidase – (+)culture will turn blue within 20 seconds when placed on dry slide, (-) no color change to blue Catalase – results of catalase test, detects presence of catalase enzyme – (+) bubbles are formed from hydrogen peroxide degraded to water and gaseous oxygen – organism has catalase; (-) no bubbles formed, organism does not have catalase Methyl red (MR) – results of methyl red test; (+) methyl red indicator turns red, indicating mixed acid fermentation of glucose, (-)methyl red becomes yellow, no mixed acid fermentation Vogues-Proskauer (VP) – results of VP test; (+) broth turns red within 60 minutes after addition of Barritt’s A and B solutions due to production of acetoin from glucose, organism uses 2,3-butanediol fermentation; (-) = broth is not red or is copper, no acetoin produced, organism does not use 2,3- butanediol fermentation Sulfur reduction (SIM) – result from SIM medium, a black precipitate in the tube indicates that sulfur has been reduced; (+) black precipitate is present, organism reduces sulfur; (-) black precipitate is absent, organism does not reduce sulfur Indole production (SIM) – result from SIM medium, after Kovac’s reagent is added, the presence of indole will cause the Kovac’s reagent to turn red/pink; (+) reagent turns pink, tryptophan is metabolized and indole is produced; (-) reagent does not turn pink, indole is not produced SIM (IndoleProduction) SIM (SulfurReduction) Phenol RedMannitol Phenol RedSucrose Phenol RedGlucose Phenol RedLactose Phenol Thioglycollate Broth Thioglycollate Oxidase DrySlide Oxidase Voges-Proskauer Simmons Citrate Nutrient gelatin MAC (growth) MSA (growth) KOH (stringy) PEA (growth) SIM (Motility) Starch Plate Endospores Methyl Red Gram Stain Catalase Shape

Family Bacillaceae fa A A A + + + + + + + + + + + + b - - - - - Bacillus cereus V V A A A + + + + + + + + + + b ------Bacillus megaterium A A A V + + + + + + + + + + + b ------Bacillus subtilis

Family Paenibacillaceae AG AG AG AG fa V + + + + + + b ------Paenibacillus polymyxa A A A + + + + + + + + b a ------Brevibacillus brevis

Family Corynebacteriaceae fa A A + + + b ------Corynebacterium xerosis fa + + + + b ------Corynebacterium. pseudodiphtheriticum

Family Lactobacillaceae AG AG fa w V A + + b ------Lactobacillus casei fa w A A + + b ------Lactobacillus delbrueckii AG AG fa w A A + + b ------Lactobacillus plantarum SIM (Indole Production) SIM (Indole SIM (SulfurReduction) Phenol RedMannitol Phenol Phenol RedSucrose Phenol RedGlucose Phenol Phenol RedLactose Thioglycollate Broth Thioglycollate Oxidase DrySlide Oxidase Voges-Proskauer Simmons Citrate Nutrient gelatin MAC (growth) MSA (growth) KOH (stringy) PEA (growth) SIM (Motility) Starch Plate Endospores Methyl Red Gram Stain Catalase Shape

Family Micrococcaceae + + + a c ------Kocuria rosea + + + + + a c ------Kocuria varians V + + + + a c ------Micrococcus luteus

Family Staphylococcaceae fa A V A V + + + + + c ------Staphylococcus capitis fa A V A + + + + + c ------Staphylococcus epidermidis fa A A A A + + + + + + c ------Staphylococcus saprophyticus

Family Streptococcaceae fa V V A A + + c ------Lactococcus lactis fa A A A A + + + + c ------Streptococcus mutans fa A V A + + + + c ------Streptococcus salivarius SIM (Indole Production) SIM (Indole SIM (SulfurReduction) Phenol RedMannitol Phenol Phenol RedSucrose Phenol RedGlucose Phenol Phenol RedLactose Thioglycollate Broth Thioglycollate Oxidase DrySlide Oxidase Voges-Proskauer Simmons Citrate Nutrient gelatin MAC (growth) MSA (growth) KOH (stringy) PEA (growth) SIM (Motility) Starch Plate Endospores Methyl Red Gram Stain Catalase Shape

Family Alcaligenaceae + + + + + + + + a b ------Achromobacter xylosoxidans + + + + + + a b ------Alcaligenes faecalis + + + + + + + b a ------Alcaligenes latus

Family Enterobacteriaceae AG AG AG AG fa B + + + + + + + ------Citrobacter freundii AG AG AG fa + + + + b ------Citrobacter rodentium AG AG AG AG fa V + + + + + + b ------Enterobacter cloacae AG fa B A + + + + + + + ------Erwinia amylovora AG fa V V A + + + + + + b ------Escherichia coli ATCC 11775 AG AG AG fa + + + + + + b ------Serratia fonticola AG fa A A A + + + + + + + + b ------Serratia liquefaciens fa A A A V + + + + + + + b ------Serratia marcescens