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What organisms require 42C incubation? Campylobacter jejuni & Pseudomonas aeruginosa What media, with temp and atmospheric requirements, are used to incubate Campylobacter jejuni? "Campy" blood agar or Skirrow medium; 42C; microaerobic What is the Gram stain result for Pseudomonas aeruginosa? Gram negative rod Can Pseudomonas aeruginosa ferment glucose? No Can Pseudomonas aeruginosa oxidize glucose? Yes Is Pseudomonas aeruginosa aerobic, anaerobic, or facultatively anaerobic? Aerobic (can adapt to anaerobic conditions) Is E. coli aerobic, anaerobic, or facultatively anaerobic? Facultative anaerobe Is Staph aureus aerobic, anaerobic, or facultatively anaerobic? Facultative anaerobe Is Listeria aerobic, anaerobic, or facultatively anaerobic? Facultative anaerobe Are yeasts aerobic, anerobic, or facultatively anaerobic? Facultative anaerobe Is Bacteroides fragilis aerobic, anaerobic or facultatively anaerobic? Anaerobic Is Prevotella aerobic, anaerobic, or facultatively anaerobic? Anaerobic Is Porphyromonas aerobic, anaerobic, or facultatively anaerobic? Anaerobic Is Fusobacterium aerobic, anaerobic, or facultatively anaerobic? Anaerobic Is Peptostreptococcus aerobic, anaerobic, or facultatively anerobic? Anaerobic Is Veilonella aerobic, anaerobic, or facultatively anerobic? Anaerobic Is Propionibacterium acnes aerobic, anaerobic, or facultatively anerobic? Anaerobic Is Actinomyces aerobic, anaerobic, or facultatively anerobic? Anaerobic Is Eubacterium aerobic, anaerobic, or facultatively anerobic? Anaerobic Is Bifidobacterium aerobic, anaerobic, or facultatively anerobic? Anaerobic Is Clostridium aerobic, anaerobic, or facultatively anerobic? Anaerobic What temperature is used to isolate Mycobacterium fortuitum-chelonae? Grows at both 28 and 37 degrees Celsius What temperature is used to incubate Mycobacterium tuberculosis? 35 to 37 degrees Celsius What temperature is used to isolate Mycobacterium marinum, M. haemophilum, & M. ulcerans? 25 to 33 degrees Celsius What temperature is used to incubate fungal cultures? 25 to 30 degrees Celsius Which Campylobacter can grow at 25 degrees Celsius? Campylobacter fetus What is the required incubation time and temperature for Yersinia enterocolitica? 22 to 25 degrees Celsius (Room temperature) for 48 hours What media and temperature is used to incubate E. coli O157:H7? Sorbitol-MacConkey (SMAC) agar; 37 degrees Celsius What media and incubation conditions are used to culture Neisseria gonorrhoaea? BAP, CHOC, & GC selective media (MTM, NYC, M-L) in CO2 at 35 degrees Celsius KIA: A/A (Yellow/Yellow) Glucose positive, Lactose positive KIA: K/A (Red/Yellow) Glucose positive, Lactose negative KIA: K/K (Red/Red) Glucose negative, Lactose negative Which organisms are KIA A/A (Yellow/Yellow)? -Enterobacter -Klebsiella -E. coli Which organisms are KIA A/A, H2S (Yellow/Black)? Citrobacter Which organisms are KIA K/A (Red/Yellow)? -Providencia -Morganella -Serratia marcescens -Shigella -Yersinia enterocolitica Which organisms are KIA K/A, H2S (Red/Black)? -Proteus -Salmonella Which organisms are KIA K/K (Red/Red)? Pseudomonas aeruginosa & Acinetobacter If an organism is Sucrose positive, Lactose negative, what would it look like on TSI? A/A (Yellow/Yellow) If an organism is Sucrose positive, Lactose negative, what would it look like on KIA? K/A (Red/Yellow) What 3 organisms are Sucrose positive and Lactose negative (TSI A/A Yellow/Yellow)? -Serratia marcescens -Yersinia enterocolitica -Proteus vulgaris If an organism is Sucrose positive & Lactose positive, what would it look like on KIA & TSI? A/A (Yellow/Yellow) If an organism is Sucrose negative & Lactose negative, what would it look like on KIA & TSI? K/A (Red/Yellow) If an organism is Sucrose negative & Lactose positive, what would it look like on KIA & TSI? A/A (Yellow/Yellow) LIA: K/K (Purple/Purple) Lysine Decarboxylase Positive LIA: K/K, H2S (Purple/Black) Lysine Decarboxylase Positive with H2S LIA: R/A (Red/Yellow) Lysine Deaminase Positive LIA: K/A (Purple/Yellow) Lysine Decarboxylase Negative Which enteric organisms are Lysine Decarboxylase Positive (LIA: Purple/Purple)? Serratia, Klebsiella, & E. Coli Which enteric organism is Lysine Decarboxylase Positive with H2S (LIA: Purple/Black)? Salmonella Which enteric organisms are Lysine Deaminase Positive (LIA: Red/Yellow)? -Proteus -Providencia -Morganella Which enteric organisms are Lysine Decarboxylase Negative (LIA: Purple/Yellow)? Enterobacter & Shigella Requires X & V Factor -Gamma Hemolytic -Porphyrin: No Fluorescence Haemophilus influenzae -Requires X & V Factor -Beta Hemolytic -Porphyrin: No Fluorescence Haemophilus haemolyticus Requires V Factor -Gamma Hemolytic -Porphyrin: Fluorescene Haemophilus parainfluenzae -Requires V Factor -Beta Hemolytic -Porphyrin: Fluorescence Haemophilus parahaemolyticus Catalase: Bubbles Positive Which organisms are Catalase positive? Staphylococcus Slide Coag: Clumping Staph aureus Slide Coag: No Clumping Perform Tube Coagulase Test Tube Coag: Clot formation Staph aureus Tube Coag: No clotting Coag-negative Staph species Latex agglutination: Clumping Staph aureus Latex Agglutination: No Clumping Coag-negative Staph species If an organism is Gram-positive and isolated from a urine sample, what additional test must be performed? Novobiocin Susceptibility Novobiocin: Susceptible Staph epidermidis (if isolated from a catheter); otherwise Coag-negative Staph Novobiocin: Resistant Staph saprophyticus Which organism isolated from a blood culture is Gram-positive, Catalase positive, and Coagulase negative? Staph lugdunensis Is it appropriate to prepare a Gram Stain of a throat culture when suspecting Group A Strep? No Is it appropriate to prepare a Gram Stain of a stool sample when suspecting Salmonella or Shigella? No Is it appropriate to prepare a Gram Stain of a throat or rectal sample when suspecting Neisseria gonorrheae? No Is it appropriate to prepare a culture of a throat or rectal sample when suspecting Neisseria gonorrheae? Yes Is it appropriate to prepare a culture of a urine or sputum specimen when suspecting an anaerobic organism? No What types of culture plates would you set up for a joint specimen? -Blood Agar Plate -Chocolate Agar -Broth What types of culture plates would you set up for a sputum specimen? -Blood Agar Plate -Chocolate Agar -EMB/MacConkey Agar What types of culture plates would you set up for a urine specimen? Blood Agar Plate & EMB/MacConkey Agar What types of culture plates would you set up for a wound specimen? -Blood Agar Plate -EMB/MacConkey Agar -PEA/CNA Agar What are common contaminants of blood culture bottles? -Coag-negative Staph -Bacillus species -Corynebacteria -Propionibacterium acnes What normal flora organisms are significant if isolated from a blood culture? Staph aureus & E. coli What type of specimens should not be refrigerated? Cerebro Spinal Fluid (CSF) & samples from normally sterile body sites What types of specimens should be refrigerated? Sputum & Urine What normal flora can be pathogens? -Listeria -Group B Strep -Staph aureus -Neisseria meningitidis What organisms are not normal flora? -Group A Strep -Bacillus anthracis -Neisseria gonorrhoeae -Haemophilus aegyptius -Haemophilus ducreyi -Salmonella -E. coli O157:H7 What type of media do you set up if you suspect Neisseria gonorrhoeae from a genital specimen? Modified Thayer-Martin (MTM), New York City (NYC), or Martin-Lewis What type of media do you set up if you suspect Neisseria gonorrhoeae from a normall sterile body fluid specimen? BAP & CHOC What type of reaction is expected when Acinetobacter & Moraxella (non-fermenters) are inoculated to OF glucose media? Green in both tubes; open & closed (oil) What type of reaction is expected when an enteric is inoculated to OF glucose media? Yellow in both tubes; open & closed (oil) What type of reaction is expected when Pseudomonas aeruginosa is inoculated to OF glucose media? Yellow in open tube; blue-green in closed tube (oil) What is the appearance of Salmonella & Shigella on EMB & MacConkey agar? Colorless What is the appearance of Salmonella & Shigella on SS agar? Shigella is colorless; Salmonella is colorless with black centers What is the appearance of Salmonella & Shigella on HE agar? Shigella is green; Salmonella is green with black centers What is the appearance of Salmonella & Shigella on XLD agar? Shigella is red; Salmonella is red with black centers Which enteric organisms are Phenylalanine positive? -Proteus -Providencia -Morganella Which enteric organisms are Phenylalanine negative & VP positive? -Enterobacter -Serratia -Klebsiella Which enteric organisms are Phenylalanine negative & VP negative? -Salmonella -Citrobacter -E. coli -Shigella Which enteric organisms are Lactose positive? -Enterobacter -Klebsiella -Citrobacter -E. coli Which enteric organisms are H2S positive? -Proteus -Salmonella -Citrobacter Which enteric organisms are Lysine positive? -Salmonella -E. coli What Enteric organisms are non-motile? -Klebsiella -Shigella An enteric organism is Phenylalanine positive, what key tests are used to further differentiate the organism? -H2S -Indole -Citrate An enteric organism is Phenylalanine positive and H2S positive, what organism could it be? Proteus What test differentiates Proteus vulgaris from Proteus mirabilis? Indole Which Proteus is Indole positive? Proteus vulgaris Which Proteus is Indole negative? Proteus mirabilis What test differentiates Providencia from Morganella? Citrate Which Phenylalanine positive organism is Citrate positive? Providencia Which Phenylalanine positive organism is Citrate negative? Morganella An enteric organism is Phenylalanine negative, what key tests are used to further differentiate the organism? -VP -Motility -Citrate What test(s) differentiate Enterobacter and Serratia? Lactose & DNase Which enteric organism is VP positive, Lactose positive, & DNase negative? Enterobacter Which enteric organism is VP positive, Lactose negative, & DNase positive? Serratia marcescens Which Klebsiella species is Indole positive? Klebsiella oxytoca Which VP negative organisms are Citrate positive? Salmonella & Citrobacter What key test differentiates Salmonella from Citrobacter? Lysine Decarboxylase Which VP negative organism is Lysine positive? Salmonella Which VP negative organism is Lysine negative? Citrobacter Which VP negative organism is Lactose positive? E. coli Which VP negative organism is Lactose negative? Shigella Which enteric organism produces mucoid colonies on agar? Klebsiella What is the characteristic pigment of Serratia marcescens on media? Pink Which organisms exhibits a characteristic swarming motility? Proteus Which organism exhibits a characteristic "School of Fish" morphology on Gram stain? Heamophilus ducreyi Spread by 4 Fs: Food, feces, fingers, flies Shigella "Bull's Eye" colonies Yersinia enterocolitica "Rose Spots" Salmonella typhi Most Enterics are urease _______? Negative What is the urease test result for Yersinia enterocolitica? Positive What is the urease test result for Klebsiella pneumoniae? Positive What is the urease test result for Enterobacter? Positive What is the urease test result for Proteus? Positive What is the urease test result for Providencia? Positive What is the urease test result for Morganella? Positive Which organisms are Oxidase positive? -Neisseria -Pseudomonas aeruginosa -Campylobacter Which organisms are Oxidase negative? Enterics Carbohydrate Utilization: Yellow Positive Carbohydrate Utilization: Red Negative Carbohydrate Utilization: Glucose positive Neisseria gonorrhoeae Carbohydrate Utilization: -Glucose: Positive -Maltose: Positive Neisseria meningitidis Carbohydrate Utilization: -Glucose: Positive -Maltose: Positive -Lactose: Positive Neisseria lactamica Butyrate Esterase: Blue Positive Butyrate Esterase: White Negative Carbohydrate Utilization: -Butyrate Esterase: Positive -Nitrate: Positive -DNase: Positive Moraxella catarrhalis Which organisms are Gram negative cocci? -Neisseria -Moraxella catarrhalis Is Moraxella a glucose fermenter, oxidizer, or non-fermenter? Non-fermenter