Review Cards - Microbiology Flashcards
Biosafety levels - Level I - risk
minimal
Biosafety levels - Level I - types of agents
those not known to cause disease in healthy adults
Biosafety levels - Bacillus subtilis
Level I
Biosafety levels - Mycobacterium gordonae
Level I
Biosafety levels - soil microbes
Level I
Biosafety levels - Level I - precautions
-standard microbiological practices
-no special equipment
Biosafety levels - Level II - risk
moderate
Biosafety levels - Level II - types of agents
common human pathogens
Biosafety levels - Enterohemorrhagic E. coli
Level II
Biosafety levels - Salmonella
Level II
Biosafety levels - HIV
Level II
Biosafety levels - HBV
Level II
Biosafety levels - influenza
Level II
Biosafety levels - Level II - precautions
-biological safety cabinet (BSC) I or II
-PPE
-autoclave must be available
-limited access
-most micro labs fall in this category
Biosafety levels - Level III - risk
high
Biosafety levels - Level III - types of agents
those that may cause serious or lethal disease via inhalation.
-effective treatment available
Biosafety levels - Bacillus anthracis
Level III
Biosafety levels - Francisella
Level III
Biosafety levels - Brucella
Level III
Biosafety levels - Mycobacterium tuberculosis
Level III
Biosafety levels - Rickettsia rickettsii
Level III
Biosafety levels - Coxiella burnetii
Level III
Biosafety levels - mold stages of systemic fungi
Level III
Biosafety levels - Level III - precautions
-biological safety cabinet (BSC) I or II
-PPE
-autoclave must be available
-limited access
-negative air flow
-sealed windows
Biosafety levels - Level IV - risk
extreme
Biosafety levels - Level IV - types of agents
those that pose a high risk of life-threatening disease
-may be transmitted by aerosols
-no vaccine or therapy
Biosafety levels - Ebola virus
Level IV
Biosafety levels - Lassa virus
Level IV
Biosafety levels - hemorrhagic fevers
Level IV
Biosafety levels - Level IV - precautions
-class III BSC
-full-body, air-supplied positive pressure suit
-independent unit with specialized ventilation & waste management to prevent release into environment
CDC Classification of Biological Agents - Category A - priority
highest
CDC Classification of Biological Agents - Category A - spread
easily disseminated or transmitted from person to person
CDC Classification of Biological Agents - Category A - impact
high mortality, potential for major public health impact
CDC Classification of Biological Agents - Category B - priority
2nd highest
CDC Classification of Biological Agents - Category B - spread
moderately easy to disseminate
CDC Classification of Biological Agents - Category B - impact
moderate illness, low death rate
CDC Classification of Biological Agents - Category C - priority
3rd highest
CDC Classification of Biological Agents - Category C - spread
could be engineered for mass dissemination
CDC Classification of Biological Agents - Category C - impact
high morbidity/mortality, major public health impact
CDC Classification of Biological Agents - Bacillus anthracis
Category A
CDC Classification of Biological Agents - Yersinia pestis
Category A
CDC Classification of Biological Agents - Francisella tularensis
Category A
CDC Classification of Biological Agents - Clostridium botulinum toxin
Category A
CDC Classification of Biological Agents - smallpox
Category A
CDC Classification of Biological Agents - hemorrhagic fever viruses (Ebola, Marburg, Lassa, Machupo)
Category A
CDC Classification of Biological Agents - Brucella
Category B
CDC Classification of Biological Agents - Salmonella
Category B
CDC Classification of Biological Agents - Shigella
Category B
CDC Classification of Biological Agents - E.coli 0157:H7
Category B
CDC Classification of Biological Agents - Burkholderia mallei
Category B
CDC Classification of Biological Agents - Burkholderia pseudomallei
Category B
CDC Classification of Biological Agents - Chlamydia psittaci
Category B
CDC Classification of Biological Agents - Coxiella burnetii
Category B
CDC Classification of Biological Agents - Clostridium perfringens toxin
Category B
CDC Classification of Biological Agents - ricin toxin
Category B
CDC Classification of Biological Agents - staphylococcal enterotoxin B
Category B
CDC Classification of Biological Agents - Rickettsia prowazekii
Category B
CDC Classification of Biological Agents - viral encephalitis viruses
Category B
CDC Classification of Biological Agents - Vibrio cholerae
Category B
CDC Classification of Biological Agents - Cryptosporidium parvum
Category B
CDC Classification of Biological Agents - Nipah virus
Category C
CDC Classification of Biological Agents - hantavirus
Category C
Biological Safety Cabinets - open front; unsterilized room air enters; air passes through high efficiency particulate air (HEPA) filter before being exhausted - class of BSC?
Class I
Biological Safety Cabinets - laminar flow cabinets with variable sash opening; air passes through 1 HEPA filter before reaching work surface & 2nd one before being exhausted - class of BSC?
Class II
Biological Safety Cabinets - completely enclosed; negative pressure; air is filter sterilized comin in & going out; gloves are attached to front - class of BSC?
Class III
Which class of BSC is most commonly used in hospital micro labs?
Class II
Which class of BSC provides minimal personnel protect and doesn’t protect work surface?
Class I
Which class of BSC provides protection for the worker & work?
Class II
Which class of BSC provides maximum protection and is used in labs that work with extremely hazardous organisms?
Class III
Sterilization & Disinfection - Temperature - steam under pressure
-autoclave
-most widely utilized in clinical labs
-15 lb of pressure for 15 min (for media)
-121C/250F
-Kills spores
-may not kill prions
Sterilization & Disinfection - Temperature - boiling
-not reliable
-spores may not be killed
Sterilization & Disinfection - Temperature - pasteurization
-used in food industry to kill foodborne pathogens
-doesn’t sterilize
-liquid is heated to 71.7*C for 15 minutes
Sterilization & Disinfection - Temperature - hot air sterilization
-used when steam may damage or fail to penetrate
-2 hours at 170*C
-kills spores
Sterilization & Disinfection - Temperature - incineration
-used to sterilize inoculating loops & biomedical wastes
Sterilization & Disinfection - Filtration
-used to sterilize liquids that are thermolabile (e.g., urea broth)
-HEPA air filters are used in BSC
Sterilization & Disinfection - Radiation
UV light wavelengths in the 200-320 nm range are used for surface disinfection both in the lab and patient rooms
Sterilization & Disinfection - Chemicals - alcohols
-ethyl & isopropyl (70-80%) frequently used as antiseptics & disinfectant
-does NOT kill spores
Sterilization & Disinfection - Chemicals - chlorine compounds
-sodium hypochlorite (household bleach)
-one of the most effective agents against HIV & HBV
-10% solution
-CAN kill spores
Sterilization & Disinfection - Chemicals - Glutaraldehyde
-effective against most vegetative cells other than mycobacteria
-used for cold sterilization of items damaged by heat, e.g., inhalation therapy equipment, equipment with optical lenses such as endoscopes
Sterilization & Disinfection - Chemicals - Hydrogen peroxide
3%-6% solution used as disinfectant
Sterilization & Disinfection - Chemicals - iodine & iodophors
-iodophors (e.g., providone-iodine) release iodine slowly & are less irritating & nonstaining
-effective skin disinfectant
-used with alcohol on skin prior to collection of blood for blood culture
Sterilization & Disinfection - Chemicals - Phenolic compounds
0.5%-3% solution for disinfecting
-does NOT kill spores
Sterilization & Disinfection - Chemicals - Quaternary ammonium compounds
-effective against wide range of vegetative bacteria
-NOT effective against spores, mycobacteria, or non-enveloped viruses
-used to disinfect floors, walls, furniture
Sterilization & Disinfection - Gases - ethylene oxide
widely used in hospitals to sterilize materials that can’t withstand steam
Anaerobes - preservative or transport device
Aspirates preferred to swabs.
Anaerobic transport system to eliminate O2.
Anaerobes - storage temperature (unpreserved)
25*C
do not refrigerate
CSF - preservative or transport device
None
CSF - storage temperature (unpreserved)
Storage not recommended, process immediately
Neisseria gonorrhoeae - preservative or transport device
Amies transport medium with charcoal, JEMBEC plates, Gono-Pak, BioBag (best to inoculate medium directly)
Neisseria gonorrhoeae - storage temperature (unpreserved)
25*C
don’t refrigerate
Sputum - preservative or transport device
None
Sputum - storage temperature (unpreserved)
4*C
Stool for culture - preservative or transport device
Transport media - e.g., Cary-Blair transport medium (has high pH and sodium thioglycollate added) I’d specimen can’t be cultured within 2 hrs of collection
Stool for culture - storage temperature (unpreserved)
4*C
-best to process without delay
-refrigeration may kill Shigella
Stool for ova & parasites - preservative or transport device
Depending on procedures to be performed: polyvinyl alcohol (PVA), 10% formalin, sodium acetate-acetic acid-formalin (SAF), merthiolate-iodine-formalin (MIF), others without formaldehyde or mercury (e.g., Ecofix, Parasafe)
Stool for ova & parasites - storage temperature (unpreserved)
Can hold formed specimens at 4*C
-soft or liquid specimens should be examined immediately or preserved
Urine - preservative or transport device
Transport tubes with boric acid-glycerol if specimen can’t be processed within 2 hrs of collection
Urine - storage temperature (unpreserved)
4*C for not more than 24 hrs
Viruses - preservative or transport device
Collect during acute phase of infection (2-3 days)
-use viral transport medium (VTM)
Viruses - storage temperature (unpreserved)
4C
-for delay >24 hrs, freeze at -70C
Criteria for rejection of specimens in microbiology - unlabeled or improperly labeled specimen - reject?
reject
Criteria for rejection of specimens in microbiology - improper collection site - reject?
reject
Criteria for rejection of specimens in microbiology - prolonged transit time (over 2 hrs without preservation) - reject?
reject
Criteria for rejection of specimens in microbiology - improper temperature during transport or storage - reject?
reject
Criteria for rejection of specimens in microbiology - leaking specimens - reject?
reject
Criteria for rejection of specimens in microbiology - specimens in non-sterile containers - reject?
reject
Criteria for rejection of specimens in microbiology - dry swab - reject?
reject
Criteria for rejection of specimens in microbiology - improper swab, e.g., wood or calcium alginate for viruses or Chlamydia - reject?
reject
Criteria for rejection of specimens in microbiology - syringes with needles attached - reject?
reject
Criteria for rejection of specimens in microbiology - culture for anaerobes requested on inappropriate sources or not received in anaerobic transport tube - reject?
reject
Criteria for rejection of specimens in microbiology - specimens received in formalin - reject?
reject
Criteria for rejection of specimens in microbiology - saliva instead of sputum - reject?
reject
Criteria for rejection of specimens in microbiology - foley catheter tip - reject?
reject
Criteria for rejection of specimens in microbiology - insufficient quantity - reject?
reject
Criteria for rejection of specimens in microbiology - formed stool for C. difficile toxin testing - reject?
reject
Criteria for rejection of specimens in microbiology - swab for acid-fast bacilli (AFB) or fungal smear and culture - reject?
reject
List organisms that frequently require special collection and processing.
- Anaerobes
- Chlamydia
- Haemophilus influenzae
- Neisseria gonorrhoeae
- Neisseria meningitidis
- Salmonella
- Shigella
- Shigella
- Streptococcus pneumoniae
- Viruses
- Parasites
Gram stain - reagent - crystal violet:
-Function?
-Outcome?
Function: primary dye
Outcome: stains all bacteria purple
Gram stain - reagent - Iodine:
-Function?
-Outcome?
Function: mordant
Outcome: reacts with crystal violet to form a complex that incorporates into peptidoglycan
Gram stain - reagent - 95% ethyl alcohol or acetone or combination:
-Function?
-Outcome?
Function: decolorizer
Outcome: removes crystal violet from gram-negative (neg) bacterial cells
Gram stain - reagent - Safranin:
-Function?
-Outcome?
Function: counterstain
Outcome: stains gram-negative bacterial cells pink
Staining properties of gram-positive & gram negative bacteria - Gram Positive - cell wall
Thick peptidoglycan layer with teichoic acid & lipoteichoic acid.
-teichoic acid cross-links prevent decolorization in Gram stain
Staining properties of gram-positive & gram negative bacteria - Gram Positive:
Stained by?
Color in gram stain?
Stained by crystal violet.
Color in gram stain: purple
Staining properties of gram-positive & gram negative bacteria - Gram Negative - cell wall
Thin peptidoglycan layer covered with proteins, phospholipids, & lipopolysaccharides.
-decolorizing agent causes increased permeability of lipid-rich cell wall
-primary stain (crystal violet) washes out
Staining properties of gram-positive & gram negative bacteria - Gram Negative - cell wall
Thin peptidoglycan layer covered with proteins, phospholipids, & lipopolysaccharides.
-decolorizing agent causes increased permeability of lipid-rich cell wall
-primary stain (crystal violet) washes out
Staining properties of gram-positive & gram negative bacteria - Gram Negative:
Stained by?
Color in gram stain?
Stained by safranin.
Color in gram stain: pink
Types of culture media - nutritive
-supports growth of most non-fastidious bacteria
Types of culture media - nutrient agar
nutritive
Type of culture media - trypticase soy agar
nutritive
Type of culture media - sheep blood agar (SBA)
enrichment
Type of culture media - chocolate agar (CHOC)
enrichment
Type of culture media - brain-heart infusion
enrichment
Type of culture media - buffered charcoal-yeast extract (BCYE) agar
enrichment
Type of culture media - Columbia colistin-nalidixic acid (CNA) agar
selective
Type of culture media - eosin methylene blue (EMB)
selective and differential
Type of culture media - MacConkey (MAC)
selective and differential
Type of culture media - Hektoen enteric (HE)
selective and differential
Type of culture media - xylose lysine deoxychocolate (XLD)
selective and differential
Type of culture media - Thayer-Martin
selective
Type of culture media - enrichment
-contains added growth factors, e.g., blood, vitamins, yeast extract
Type of culture media - selective
-contains additives such as dyes, bile salts, alcohols, acids, or antibiotics to inhibit growth of certain bacteria (e.g., gram pos)
Type of culture media - differential
-contains compounds that allow certain bacteria (or even species) to be visually differentiated (e.g., lactose fermentation, hydrogen sulfide [H2S], production)
Routine media for aerobes and facultative anaerobes - sheep blood agar (SBA) - type?
-enrichment
-differential
Routine media for aerobes and facultative anaerobes - sheep blood agar (SBA) - for isolation of?
-most non-fastidious bacteria
Routine media for aerobes and facultative anaerobes - sheep blood agar (SBA):
Composed of?
Allow differentiation of?
tryptic soy agar with 5% sheep blood
-allows differentiation of hemolysis
Routine media for aerobes and facultative anaerobes - chocolate agar (CHOC) - type?
enrichment
Routine media for aerobes and facultative anaerobes - chocolate agar (CHOC) - for isolation of?
fastidious organisms including Haemophilus & Neisseria
Routine media for aerobes and facultative anaerobes - chocolate agar (CHOC):
Supplies what factors?
Incubated?
supplies X (hemin) & V (NAD) factors
-incubate in 5% CO2
Routine media for aerobes and facultative anaerobes - Columbia colistin-nalidixic acid (CNA) agar - type?
selective
Routine media for aerobes and facultative anaerobes - Columbia colistin-nalidixic acid (CNA) agar - for isolation of?
gram-positive cocci (GPC), especially from wound & stool culture
Routine media for aerobes and facultative anaerobes - Columbia colistin-nalidixic acid (CNA) agar - what does colistin & nalidixic acid do?
Colistin & nalidixic acid suppress most gram-negative (GN)
-contains 5% sheep blood but should not be used to observe hemolytic reactions.
Routine media for aerobes and facultative anaerobes - Phenylethyl alcohol agar (PEA) - type?
selective
Routine media for aerobes and facultative anaerobes - Phenylethyl alcohol agar (PEA) - for isolation of?
gram-positive cocci (GPC) & anaerobic gram-negative rods (GNR)
Routine media for aerobes and facultative anaerobes - Phenylethyl alcohol agar (PEA) - what does phenylethyl alcohol do?
Phenylethyl alcohol inhibits enteric GNR
-contains 5% sheep blood
Routine media for aerobes and facultative anaerobes - brain heart infusion agar - type?
enrichment & selective
Routine media for aerobes and facultative anaerobes - brain heart infusion agar - for isolation of?
fastidious and non-fastidious organisms (anaerobes included)
Routine media for aerobes and facultative anaerobes - brain heart infusion agar - useful for culturing?
useful for culturing streptococci, pneumococci, and meningococci
Routine media for aerobes and facultative anaerobes - brain heart infusion agar - what can be added to detect VRE?
vancomycin
VRE = vancomycin-resistant enterococci
Routine media for aerobes and facultative anaerobes - eosin methylene blue (EMB) - type?
selective & differential
Routine media for aerobes and facultative anaerobes - eosin methylene blue (EMB) - for isolation of?
enteric gram negative rods (GNR)
Routine media for aerobes and facultative anaerobes - eosin methylene blue (EMB):
Function of eosin and methylene blue?
Lactose fermenters?
E.coli?
Non-lactose fermenter?
-eosin & methylene blue inhibit gram positive (GP)
-LF = green-black or purple
-E.coli = green metallic sheen
-NLF = colorless or light purple
Routine media for aerobes and facultative anaerobes - MacConkey (MAC) agar - type?
selective & differential
Routine media for aerobes and facultative anaerobes - MacConkey (MAC) agar - for isolation of?
enteric gram negative rods (GNR)
Routine media for aerobes and facultative anaerobes - MacConkey (MAC) agar:
-Function of bile salts & crystal violet?
-Lactose fermenters?
-Non-lactose fermenters?
-bile salts & crystal violet inhibits most GP
-LF = pink
-NLF = colorless
Routine media for aerobes and facultative anaerobes - Sorbitol MacConkey (SMAC) agar - type?
selective
Routine media for aerobes and facultative anaerobes - Sorbitol MacConkey (SMAC) agar - for isolation of?
E. coli 0157:H7
Routine media for aerobes and facultative anaerobes - Sorbitol MacConkey (SMAC) agar:
-E. coli 0157:H7?
-Doesn’t ferment sorbitol = colorless colonies
-Some labs have stopped using because non-0157 serotypes can be pathogens
Routine media for aerobes and facultative anaerobes - Hektoen enteric (HE) - type?
selective & differential
Routine media for aerobes and facultative anaerobes - Hektoen enteric (HE) - for isolation of?
Salmonella & Shigella in stool
Routine media for aerobes and facultative anaerobes - Hektoen enteric (HE):
-Function of bile salts, bromothymol blue, & acid fusion?
-Nonpathogens?
-Non-lactose fermenters?
-H2S positive?
-Bile salts, bromothymol blue, & acid fuchsin inhibit normal GI flora
-Nonpathogens = orange to salmon pink
-NLF = green to blue-green
-H2S pos colonies = black precipitate
Routine media for aerobes and facultative anaerobes - xylose lysine deoxycholate (XLD) - type?
selective & differential
Routine media for aerobes and facultative anaerobes - xylose lysine deoxycholate (XLD) - for isolation of?
Salmonella & Shigella in stool
Routine media for aerobes and facultative anaerobes - xylose lysine deoxycholate (XLD):
-Function of deoxycholate?
-4 types of colonies?
-Deoxycholate inhibits many GNR & GP
-4 types of colonies:
1. yellow (e.g., E. coli)
2. yellow with black centers (e.g., some Proteus species)
3. colorless or red colonies (e.g., Shigella)
4. red colonies with black centers (e.g., Salmonella)
-Some shigellae may be inhibited.
-Some salmonellae may not produce H2S
Routine media for aerobes and facultative anaerobes - Salmonella-Shigella (SS) agar - type?
selective
Routine media for aerobes and facultative anaerobes - Salmonella-Shigella (SS) agar - for isolation of?
Salmonella & Shigella in stool
Routine media for aerobes and facultative anaerobes - Salmonella-Shigella (SS) agar:
-Function of brilliant green & bile salts?
-Ferment lactose?
-Produce H2S (black center)?
-brilliant green & bile salts inhibit other enterics.
-Salmonella & Shigella DO NOT ferment lactose - colorless colonies
-Salmonella produces H2S (black center)
Routine media for aerobes and facultative anaerobes - Deoxycholate-citrate agar - type?
selective
Routine media for aerobes and facultative anaerobes - Deoxycholate-citrate agar - for isolation of?
Salmonella & Shigella
Routine media for aerobes and facultative anaerobes - Deoxycholate-citrate agar - inhibits?
other non-pathogenic enterics
Routine media for aerobes and facultative anaerobes - Thioglycolate medium - type?
enrichment
Routine media for aerobes and facultative anaerobes - Thioglycolate medium - for isolation of?
Campylobacter from stool, wound cultures
Routine media for aerobes and facultative anaerobes - Thioglycolate medium - subculture to?
Campy-selective agar after overnight incubation at 4*C
Routine media for aerobes and facultative anaerobes - Campylobacter blood agar (Campy BAP) - type?
Enrichment & selective
Routine media for aerobes and facultative anaerobes - Campylobacter blood agar (Campy BAP) - for isolation of?
Campylobacter from stool
Routine media for aerobes and facultative anaerobes - Campylobacter blood agar (Campy BAP) - incubation?
Incubate plates in 5% CO2 at 42*C
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - Modified Thayer-Martin (MTM) - composed of?
Vancomycin
Colistin
Nystatin
Trimethoprim (TMP)
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - Modified Thayer-Martin (MTM) - function of Vancomycin, colistin, nystatin, and trimethoprim?
inhibit growth of normal genital flora
-some N. gonorrhoeae may be inhibited
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - Modified Thayer-Martin (MTM) - incubation?
incubate in increased CO2
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - Martin-Lewis - similar to?
similar to Thayer-Martin, but different antibiotics - inhibits YEAST better
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - Martin-Lewis - incubation?
incubate in increased CO2
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - New York City medium (NYC) - incubation?
incubate in increased CO2
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - New York City medium (NYC) - inhibited? Growth?
Some N. gonorrhoeae are inhibited by antibiotics.
Genital mycoplasmas will grow.
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - GC-LECT - antibiotics?
Contains antibiotics to inhibit gram positive & gram negative bacteria and yeast
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - JEMBEC plates - function?
For transportation & growth of N. gonorrhoeae
Selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis - JEMBEC plates - contain?
Plates contain Neisseria-selective medium & come with resealable polyethylene bag & CO2-generating tablet.
–NO need to transfer to a culture plate.
List the selective media for isolation of Neisseria gonorrhoeae and Neisseria meningitidis.
- Modified Thayer-Martin (MTM)
- Martin-Lewis
- New York City medium (NYC)
- GC-LECT
- JEMBEC plates
Special bacteriologic media - Cystine-tellurite blood agar - use?
Differential medium for isolation of Corynebacterium diphtheriae
Special bacteriologic media - Cystine-tellurite blood agar - colonies on agar?
C. diphtheriae produces dark grey to black colonies.
Special bacteriologic media - Loeffler medium - use?
Enrichment medium for C. diphtheriae
Special bacteriologic media - Loeffler medium - function?
promotes development of metachromatic granules
Special bacteriologic media - Tindale agar - use?
selective differential medium for C. diphtheriae
Special bacteriologic media - Tindale agar - colonies?
Corynebacterium spp. produce gray to black colonies due to reduction of tellurite.
-C. diphtheriae colonies are surrounded by a brown halo.
Special bacteriologic media - Bismuth sulfite agar:
-selective for?
-function of bismuth sulfite & brilliant green?
-selective for Salmonella
-inhibits most others
Special bacteriologic media - Bismuth sulfite agar - S. typhi colonies?
S. typhi colonies are black, surrounded by metallic sheen.
-others are light green
-Some salmonellae may be inhibited
Special bacteriologic media - Cefsulodin-Irgasan-novobiocin (CIN) agar - use?
Selective medium for Yersinia enterocolitica, Aeromonas, & Plesiomonas shigelloides
Special bacteriologic media - Cefsulodin-Irgasan-novobiocin (CIN) agar:
-Function of crystal violet?
-Function of novobiocin?
-Function of Cefsulodin?
-Crystal violet inhibits most gram negatives.
-Novobiocin inhibits gram positive cocci
-Cefsulodin inhibits most gram positives & gram negatives
Special bacteriologic media - Cefsulodin-Irgasan-novobiocin (CIN) agar - Y. enterocolitica colonies?
Y. enterocolitica ferments mannitol, appears as red “bull’s-eye” colonies surrounded by colorless halo.
Special bacteriologic media - Alkaline peptone water (APW) - use?
Enrichment media for recovery of Vibrio from stool
Special bacteriologic media - Alkaline peptone water (APW):
-Function of alkaline pH?
-Subculture to?
-Alkaline pH suppresses commensals
-Subcultured to thiosulfate citrate bile salts sucrose (TCBS)
Special bacteriologic media - Thiosulfate Citrate Bile Salts Sucrose (TCBS) agar - use?
Selective for Vibrio
Special bacteriologic media - Thiosulfate Citrate Bile Salts Sucrose (TCBS) agar - function of high pH?
inhibits most bacteria
Special bacteriologic media - Thiosulfate Citrate Bile Salts Sucrose (TCBS) agar:
-V. cholerae colonies?
-V. parahaemolyticus and V. vulnificus colonies?
-V. cholerae ferments sucrose, produces yellow colonies
-V. parahaemolyticus & V. vulnificus DO NOT ferment sucrose; usually produce blue-green colonies
Special bacteriologic media - Bordet-Gengou agar - use?
Selective enrichment medium for isolation of Bordetella pertussis
Special bacteriologic media - Bordet-Gengou agar:
-composed of?
-function of methicillin?
-Potato-glycerol-based medium enriched with blood.
-Contaminants inhibited by methicillin.
Special bacteriologic media - Bordet-Gengou agar - Bordetella colonies?
Bordetella colonies resemble mercury droplets.
Special bacteriologic media - Bordet-Gengou agar - what is the plate also known as?
“Cough plate.”
Special bacteriologic media - Regan-Lowe agar - use?
Selective for B. pertussis
Special bacteriologic media - Regan-Lowe agar - composed of?
Charcoal agar supplemented with horse blood, cephalexin, & amphotericin B.
Special bacteriologic media - BYCE agar - use?
Enrichment medium for isolation of Legionella
Special bacteriologic media - BYCE agar:
-Function of yeast extract & L-cysteine?
-Function of charcoal?
-Yeast extract & L-cysteine enhance growth of Legionella.
-Charcoal absorbs toxic compounds.
Special bacteriologic media - V agar - use?
Selective & differential for G. vaginalis
Special bacteriologic media - V agar - incubation?
Incubate in increased CO2 for 48 hours.
Special bacteriologic media - V agar - G. vaginalis colonies?
G. vaginalis colonies are beta hemolytic.
Special bacteriologic media - CHROMagar or chromID - use?
Detect emerging antibiotic-resistant pathogens such as extended-spectrum beta-lactamase (ESBL) - producing E.coli, methicillin-resistant Staphylococcus aureus (MRSA), and carbapenemase-producing Enterobacteriaceae
Special bacteriologic media - CHROMagar or chromID - function of chromogens?
Chromogens are released if specific enzymes produced by target organism are present.
Aerotolerance test - Aerobe:
-blood agar incubated aerobically?
-blood agar incubated anaerobically?
-CHOC incubated in CO2 incubator?
-Growth
-No growth
-Growth
Aerotolerance test - Capnophilic aerobe:
-blood agar incubated aerobically?
-blood agar incubated anaerobically?
-CHOC incubated in CO2 incubator?
-No growth
-No growth
-Growth
Aerotolerance test - Facultative anaerobe:
-blood agar incubated aerobically?
-blood agar incubated anaerobically?
-CHOC incubated in CO2 incubator?
-Growth
-Growth
-Growth
Aerotolerance test - Obligate anaerobe:
-blood agar incubated aerobically?
-blood agar incubated anaerobically?
-CHOC incubated in CO2 incubator?
-No growth
-Growth
-No growth
Aerotolerance test - Aerotolerant anaerobe:
-blood agar incubated aerobically?
-blood agar incubated anaerobically?
-CHOC incubated in CO2 incubator?
-No growth
-Growth
-Growth
List the organisms requiring incubation in increased CO2.
- Campylobacter (10-15% CO2)
- Haemophilus (5-10% CO2)
- Helicobacter (5-10% CO2)
- Moraxella catarrhalis (5% CO2)
- Mycobacterium (5-10% CO2)
- Pathogenic Neisseria (5-10% CO2)
Hemolytic reactions on sheep blood agar - Alpha hemolysis:
-description
-explanation
-examples
-green zone around colony; may be narrow or wide
-partial lysis of RBCs
-Streptococcus pneumoniae & viridans streptococci
Hemolytic reactions on sheep blood agar - Beta hemolysis:
-description
-explanation
-examples
-clear zone around colony; may be narrow or wide
-complete lysis of RBCs
-Group A strep (GAS), group B strep (GBS), Listeria monocytogenes
Hemolytic reactions on sheep blood agar - Gamma (non-hemolytic):
-description
-explanation
-examples
-no zone of hemolysis
-no lysis of RBCs
-Enterococcus faecalis
Staphylococcaceae - Staphylococcus spp. - gram stain
gram positive cocci - usually in clusters
Staphylococcaceae - Staphylococcus spp. - colonies of SBA
1-3 mm
round
smooth
convex
glistening
opaque
entire edge
butyrous (butter-like)
Staphylococcaceae - Staphylococcus spp. - key characteristics
-catalase positive
-fermentative
-modified oxidase (microdase) negative
-resistant to bacitracin
-Matrix-assisted laser desorption ionization-time of flight (MALD-TOF) mass spectrometry routinely used for ID of isolates
Staphylococcaceae - Staphylococcus spp. - normal flora of?
skin
mouth
pharynx
vagina
urethra
GI tract
Staphylococcaceae - Staphylococcus spp. - aerotolerance
facultative anaerobe
Staphylococcaceae - Staphylococcus spp. - growth on media
Grows on most non-selective media
Staphylococcaceae - Staphylococcus spp. - salt tolerance
salt tolerant
Staphylococcaceae - S. aureus - pathogenicity
Causes suppurative cutaneous infections, toxic shock syndrome, food poisoning
Staphylococcaceae - S. aureus - gram stain
gram positive cocci - usually in clusters
Staphylococcaceae - S. aureus - colonies on SBA
-most are beta hemolytic (small zone)
-may be golden
Staphylococcaceae - S. aureus - key characteristics
-coagulase positive
-ferments mannitol (yellow color with mannitol salt agar (MSA))
-usually DNase positive
Staphylococcaceae - S. aureus - penicillin
85-90% resistant to penicillin
Staphylococcaceae - S. aureus - MRSA
-strains are common
-methicillin resistance is mediated by the mecA gene or its homologs and is the most important resistance mechanism
Which staphylococcaceae species is the second leading cause of health-care-associated infections, spread by direct contact with personnel and contaminated objects?
A. S. saprophyticus
B. S. aureus
C. S. epidermidis
D. Micrococcus
B. S. aureus
Staphylococcaceae - S. epidermidis - pathogenicity
-opportunistic pathogen
-causes nosocomial infections due to contamination of inserted or implanted medical devices
Staphylococcaceae - S. epidermidis - gram stain
gram positive cocci - usually in clusters
Staphylococcaceae - S. epidermidis - colonies on SBA
-white to slightly yellow
-usually non-hemolytic
Staphylococcaceae - S. epidermidis - key characteristics
-coagulase negative
-grows on MSA but does NOT ferment mannitol
-sensitive to novobiocin
Which staphylococcaceae is frequently recovered from blood cultures but not usually clinically significant in adults?
A. S. aureus
B. S. epidermidis
C. S. saprophyticus
D. Micrococcus
B. S. epidermidis
Staphylococcaceae - S. saprophyticus - pathogenicity
-UTI in young sexually active females
-Urethritis & prostatitis in males
Staphylococcaceae - S. saprophyticus - gram stain
gram positive cocci - usually in clusters
Staphylococcaceae - S. saprophyticus - colonies on SBA
-white to slightly yellow
-non-hemolytic
Staphylococcaceae - S. saprophyticus - key characteristics
-coagulase negative
-resistant to novobiocin
-may ferment mannitol
Which of the following characteristics can help differentiate between S. epidermidis and S. saprophyticus?
A. coagulase
B. mannitol fermentation
C. novobiocin sensitivity
D. none of the above
C. novobiocin sensitivity
-S. epidermidis is sensitive to novobiocin
-S. saprophyticus is resistant to novobiocin
-*S. saprophyticus may or may not ferment mannitol; S. epidermidis does NOT ferment mannitol.
-Both are coagulase negative
Staphylococcaceae - S. saprophyticus - when is novobiocin sensitivity performed?
Only when coagulase-negative staphylococci (CNS) are isolated from the urine of a female.
Staphylococcaceae - Micrococcus - pathogenicity
-usually non-pathogenic
-found in environment and on skin & mucous membranes
Staphylococcaceae - Micrococcus - gram stain
Large gram positive cocci in pairs or tetrads (predominant arrangement)
Staphylococcaceae - Micrococcus - colonies on SBA
-often pigmented (bright yellow, orange, pink, tan)
-high-domed colonies
Staphylococcaceae - Micrococcus - key characteristics
-catalase positive
-coagulase negative
-oxidative
-Modified oxidase (microdase) positive
-most urease positive
-resistant to furazolidone & lysostaphin (in contrast to most staphylococci)
How would you differentiate Micrococcus from staphylococci?
A. coagulase
B. modified oxidase (microdase)
C. All of the above
D. None of the above
B. modified oxidase (microdase)
Staphylococcaceae - Micrococcus - aerotolerance
usually grow aerobically
Staphylococcaceae - Micrococcus - species in this genus
Micrococcus luteus
Micrococcus lylae
List the tests used in the identification of staphylococci.
- Catalase
- Coagulase
- Slide agglutination tests for S. aureus
- MSA
- Rapid latex and hemagglutinin assays
- MALDI-TOF
Tests for identification of staphylococci - Catalase - principle
Enzyme catalase converts 3% hydrogen peroxide (H2O2) to oxygen & water. Immediate bubbling.
Tests for identification of staphylococci - Catalase - key reactions
Staphylococcus & micrococcus are catalase positive
Tests for identification of staphylococci - Catalase - bubbling
Bubbles after 20-30 seconds are NOT considered a positive reaction.
Catalase from RBCs in blood agar may produce weak bubbles.
Tests for identification of staphylococci - Coagulase - principle
Enzyme coagulase causes coagulation (tube test) or agglutination (slide test) in plasma.
Tests for identification of staphylococci - Coagulase - key reactions
S. aureus is coagulase positive.
Tests for identification of staphylococci - Coagulase - slide test vs. tube test
Slide test is a screening test - detects bound coagulase (clumping factor).
-If negative, a tube test should be performed - detects free coagulase.
Largely replaced by latex agglutination tests
Tests for identification of staphylococci - Slide agglutination tests for S. aureus - principle
Agglutination of latex beads coated with fibrinogen & antibodies to protein A (protein in cell wall of S. aureus).
Tests for identification of staphylococci - Slide agglutination tests for S. aureus - key reactions
S. aureus positive
Tests for identification of staphylococci - Slide agglutination tests for S. aureus - reporting by labs
Most labs report positive organisms as S. aureus.
Tests for identification of staphylococci - Mannitol Salt agar (MSA) - principle
Fermentation of mannitol results in color change from pink to yellow.
Tests for identification of staphylococci - Mannitol Salt agar (MSA) - key reactions
S. aureus positive
Tests for identification of staphylococci - Mannitol Salt agar (MSA) - what component inhibits most organisms other than staph?
7.5% salt
Tests for identification of staphylococci - Mannitol Salt agar (MSA) - do all staph grow on MSA?
They all can grow on MSA.
Tests for identification of staphylococci - Mannitol Salt agar (MSA) - why is it rarely used for ID of S. aureus today?
because other species ferment mannitol
Tests for identification of staphylococci - Rapid latex and hemagglutination assays - principle
Latex agglutination slide-card tests to detect coagulase and/or protein A associated with S. aureus & MRSA
Tests for identification of staphylococci - Rapid latex and hemagglutination assays - key reactions
S. aureus positive
Tests for identification of staphylococci - Rapid latex and hemagglutination assays - sensitivity & specificity
High sensitivity but relatively low specificity.
-May get false-positive with coagulase-negative staph.
Tests for identification of staphylococci - MALDI-TOF - principle
A type of mass spectrometry good for analyzing biomolecules like peptides, lipids, and saccharides.
Tests for identification of staphylococci - MALDI-TOF - key reactions
Rapid and specific ID of staphylococci and other catalase-positive cocci.
Tests for identification of staphylococci - MALDI-TOF - % of identification
ID of staphylococci at species levels is >97% but lower for coagulase-negative staph.
Streptococci - Streptococcus spp. - gram stain
Oval gram positive cocci in chains & pairs
Streptococci - Streptococcus spp. - colonies on SBA
-<1mm
-white to gray
-translucent or semi-opaque
-variable hemolysis
Streptococci - Streptococcus spp. - key characteristics
Catalase negative
-differentiates for staph
Streptococci - Streptococcus spp. - aerotolerance
Facultative anaerobes
Streptococci - Streptococcus spp. - growth media
-require enriched media
-chaining best in broth cultures
Streptococci - Group A Strep (GAS) - pathogenicity
-causes 90% of strep infections
-Strep sore throat
-rheumatic fever
-glomerulonephritis
-scarlet fever
-erysipelas
-puerperal sepsis
-impetigo
Streptococci - Group A Strep (GAS) - gram stain
oval gram positive cocci in chains
Streptococci - Group A Strep (GAS) - colonies on SBA
-pinpoint
-grayish white
-translucent
-usually beta hemolytic (wide zone)
Streptococci - Group A Strep (GAS) - key characteristics
-sensitive to bacitracin
-pyrrolidonyl aminopeptidase (PYR) positive
Streptococci - Group A Strep (GAS) - most common GAS?
S. pyogenes
Streptococci - Group A Strep (GAS) - hemolysis due to?
O2-stable streptolysin S & O2-labile streptolysin O.
Streptococci - Group A Strep (GAS) - definitive diagnosis
Immunoassay for Lancefield group A or MALDI-TOF
Streptococci - Group B Strep (GBS), S. agalactiae - pathogenicity
-normal flora of female genital tract
-most common cause of neonatal septicemia & meningitis
Streptococci - Group B Strep (GBS), S. agalactiae - gram stain
oval gram positive cocci in chains
Streptococci - Group B Strep (GBS), S. agalactiae - colonies on SBA
-slightly larger than GAS
-gray-white
-narrow zone of diffuse beta hemolysis
-may be non-hemolytic
Streptococci - Group B Strep (GBS), S. agalactiae - key characteristics
-resistant to bacitracin
-sodium hippurate positive
-CAMP positive
Streptococci - Group B Strep (GBS), S. agalactiae - pregnant women
Vaginal & rectal swabs collected from pregnant women at 35-37 weeks gestation
Streptococci - Group B Strep (GBS), S. agalactiae - growth media
Inoculated in selective broth, e.g., LIM, GBS broth
Streptococci - Group B Strep (GBS), S. agalactiae - test recommended for ID after culture enrichement?
Nucleic acid amplification tests (NAATs)
Streptococci - Streptococcus pneumoniae - pathogenicity
-colonizes upper respiratory tract of some children
-most common cause of community acquired pneumonia
-major cause of otitis media & meningitis in adults
-infects sinuses and eyes
Streptococci - Streptococcus pneumoniae - gram stain
-football-shaped (lancet-shaped) gram positive cocci
-usually in pairs
-may be single or in short chains
-frequently encapsulated
Streptococci - Streptococcus pneumoniae - colonies on SBA
-round
-translucent
-glistening
-dome-shaped when young
-central depression with age (umbilicate) due to autolysis
-Alpha hemolytic
-Encapsulated strains are mucoid
Streptococci - Streptococcus pneumoniae - key characteristics
-bile solubility positive
-sensitive to optochin
Streptococci - Streptococcus pneumoniae - Lancefield group
None
Streptococci - Streptococcus pneumoniae - tests used for ID
MALDI-TOF can be used
Streptococci - Group C/G streptococci - S. dysgalactiae subsp equisimilis - pathogenicity
-normal flora of upper respiratory tract but can cause pharyngitis (throat infection)
Streptococci - Group C/G streptococci - S. dysgalactiae subsp equisimilis - gram stain
oval gram positive cocci in clusters & pairs
Streptococci - Group C/G streptococci - S. dysgalactiae subsp equisimilis - colonies on SBA
beta-hemolytic
Streptococci - Group C/G streptococci - S. dysgalactiae subsp equisimilis - key characteristics
-resistant to bacitracin
-PYR positive
Streptococci - Group C/G streptococci - S. dysgalactiae subsp equisimilis - indistinguishable from?
Indistinguishable from pharyngitis caused by S. pyogenes.
Streptococci - Viridans streptococci (S. mitis group, S. mutans group, S. salivarius group, & S. bovis group) - pathogenicity
-normal in oral, respiratory, & GI mucosa
-opportunistic pathogens
-frequent cause of subacute bacterial endocarditis, bacteremia, sepsis, and abscesses.
Streptococci - Viridans streptococci (S. mitis group, S. mutans group, S. salivarius group, & S. bovis group) - gram stain
gram positive cocci in chains
Streptococci - Viridans streptococci (S. mitis group, S. anginosus group, S. mutans group, S. salivarius group, & S. bovis group) - colonies on SBA
-alpha hemolytic (S. mitis group)
-non-hemolytic (S. salivarius and S. bovis group)
-beta-hemolytic (S. anginosus group)
Streptococci - Viridans streptococci (S. mitis group, S. mutans group, S. salivarius group, & S. bovis group) - key characteristics
-S. mutans group - ferments sorbitol
-Bile solubility negative
-S. bovis is bile esculin positive
Enterococcus - E. faecium & E. faecalis - pathogenicity
-normal in mouth, GI tract, & male genital tract
-causes nosocomial UTI, wound infections, endocarditis, & bacteremia
Enterococcus - E. faecium & E. faecalis - gram stain
oval gram positive cocci in pairs, chains
Enterococcus - E. faecium & E. faecalis - colonies on SBA
E. faecium - alpha or non-hemolytic
-Some strains of E. faecalis may be beta-hemolytic
Enterococcus - E. faecium & E. faecalis - key characteristics
-catalase negative
-hydrolyzes esculin
-grows in 6.5% NaCl broth
-PYR positive
Enterococcus - E. faecium & E. faecalis - what antibiotic is E. faecalis resistant to?
Vancomycin
-VRE are a significant public health concern.
-VRE fecal surveillance culture is often done for early detection
Enterococcus - E. faecium & E. faecalis - media used for ID of VRE
chromogenic agar
Lists the test used for the identification of beta-hemolytic streptococci.
- Bacitracin disk
- MALDI-TOF
- PYR test
- CAMP test
- Hippurate hydrolysis
- Slide agglutination tests
Tests for identification of beta-hemolytic streptococci - Bacitracin disk - principle
Zone of inhibition after overnight incubation = susceptibility to bacitracin
Tests for identification of beta-hemolytic streptococci - Bacitracin disk - key reactions
-Group A strep susceptible
-Group B strep resistant
Tests for identification of beta-hemolytic streptococci - Bacitracin disk:
-also known as?
-media performed on?
-no longer recommended - why?
-replaced by?
-also known as A disk
-performed on SBA
-no longer recommended because some groups C & G are susceptible
-replaced by PYR & serogrouping by latex agglutination
Tests for identification of beta-hemolytic streptococci - MALDI-TOF - principle
A type of mass spectrometry good for identifying biomolecules like peptides, lipids, and saccharides.
Tests for identification of beta-hemolytic streptococci - MALDI-TOF - key reactions
Reliable for S. pyogenes, S. agalactiae, & S. pneumoniae
Tests for identification of beta-hemolytic streptococci - PYR test - principle
If PYR is hydrolyzed = red color after addition of color developer
Tests for identification of beta-hemolytic streptococci - PYR test - key reactions
group A strep are positive
Tests for identification of beta-hemolytic streptococci - PYR test - more specific than what other test for group A strep?
bacitracin disk test
True or False. Group A strep is the only beta-hemolytic strep that is PYR positive.
True
Tests for identification of beta-hemolytic streptococci - CAMP test - principle
Group B strep produces extracellular protein that enhances hemolysis of beta-hemolytic S. aureus on SBA
Tests for identification of beta-hemolytic streptococci - CAMP test - key reactions
group B strep is positive
Tests for identification of beta-hemolytic streptococci - CAMP test - classic method
-unknown is streaked perpendicular to a streak of beta-lysin producing S. aureus.
-incubated in ambient air overnight
-FALSE POSITVE in CO2
-Arrowhead hemolysis - where inoculum lines meet
-Disk containing beta-lysin can be used instead of S. aureus
Tests for identification of beta-hemolytic streptococci - CAMP test - rapid test
A drop of beta-lysin on colonies on SBA.
-only requires 20 minute incubation
List the tests used for identification of beta-hemolytic streptococci.
- Hippurate hydrolysis
- Slide agglutination tests
Tests for identification of beta-hemolytic streptococci - Hippurate hydrolysis - principle
Organisms that produce hippuricase (hippurate hydrolase) hydrolyze sodium hippurate to benzoate & glycine.
Tests for identification of beta-hemolytic streptococci - Hippurate hydrolysis - key reactions
group B strep positive
Tests for identification of beta-hemolytic streptococci - Hippurate hydrolysis - alternative to?
Alternative to CAMP.
-2 hr test available
Tests for identification of beta-hemolytic streptococci - Slide agglutination tests - principle
Latex particles coated with group-specific antibodies agglutinate in the presence of bacterial antigens.
Tests for identification of beta-hemolytic streptococci - Slide agglutination tests - key reactions
Organisms agglutinate in corresponding antisera
Tests for identification of beta-hemolytic streptococci - Slide agglutination tests - tests commercially available for rapid ID of?
groups A, B, C, D, F, G strep, & S. pneumoniae
List the tests used for identification of alpha-hemolytic streptococci.
- Optochin disk
- Bile solubility
Tests for identification of alpha-hemolytic streptococci - Optochin disk - principle
Zone of inhibition >=14 mm with 6-mm disk OR >=16 mm with 10-mm disk = susceptibility to optochin
Tests for identification of alpha-hemolytic streptococci - Optochin disk - key reactions
-S. pneumoniae susceptible
-Viridans strep resistant
Tests for identification of alpha-hemolytic streptococci - Optochin disk:
-also known as?
-contains?
-method?
-also known as P disk
-contains ethylhydrocupreine hydrochloride
-Method: Placed on lawn of inoculum on SBA. Plates incubated overnight in CO2. If zone of inhibin is <14 mm, ID organisms as S. pneumoniae only if bile soluble.
Tests for identification of alpha-hemolytic streptococci - Bile solubility - principle
Bile salts (e.g., sodium deoxycholate) causes lysis of some organisms
Tests for identification of alpha-hemolytic streptococci - Bile solubility - key reactions
S. pneumoniae positive
Tests for identification of alpha-hemolytic streptococci - Bile solubility - performed on? positive?
-can be performed in broth or on colony
-POS = clearing of broth or disappearance of colony
-results in 30 seconds or less
Antibiograms of gram-positive cocci - Methicillin-sensitive Staphylococcus aureus - susceptibility
-resistant to penicillin
-susceptible to cephalosporin & vancomycin
Antibiograms of gram-positive cocci - Methicillin-sensitive Staphylococcus aureus - routine susceptibility testing?
yes
Antibiograms of gram-positive cocci - Methicillin-resistant Staphylococcus aureus (MRSA) - susceptibility
-resistant to all beta-lactams (penicillin, ampicillin, and cephalosporin)
Antibiograms of gram-positive cocci - Methicillin-resistant Staphylococcus aureus (MRSA) - routine susceptibility testing?
yes
Antibiograms of gram-positive cocci - Group A Strep (GAS) - susceptibility
-universally susceptible to penicillin, cephalosporins, vancomycin
Antibiograms of gram-positive cocci - Group A Strep (GAS) - routine susceptibility testing?
no
Antibiograms of gram-positive cocci - Group B Strep (GBS) - susceptibility
Susceptible to penicillin, cephalosporins, vancomycin
Antibiograms of gram-positive cocci - Group B Strep (GBS) - routine susceptibility testing?
variable - may be done when isolated from baby
Antibiograms of gram-positive cocci - Streptococcus pneumoniae - susceptibility
-increasingly resistant to penicillin
-Susceptible to vancomycin
Antibiograms of gram-positive cocci - Streptococcus pneumoniae - routine susceptibility testing?
yes
Antibiograms of gram-positive cocci - Enterococcus - susceptibility
E. faecium
-usually resistant to penicillin & several other commonly used antibiotics
-most are susceptible to vancomycin
*E. faecium
-vancomycin resistant (VRE)
Antibiograms of gram-positive cocci - Enterococcus - routine susceptibility testing?
yes
Aerobic spore-forming gram-positive rods - Bacillus spp. - pathogenicity
usually contaminants
Aerobic spore-forming gram-positive rods - Bacillus spp. - gram stain
-large with square ends
-may be in chains
-oval, central to subterminal spores that aren’t swollen
-looks like bamboo
-spores may not be seen in direct smear
Aerobic spore-forming gram-positive rods - Bacillus spp. - culture
-large spreading beta-hemolytic colonies with irregular edges (MEDUSA HEAD)
-whitish gray
-may be pigmented
Aerobic spore-forming gram-positive rods - Bacillus spp. - key characteristics
-catalase positive
-most are motile
Aerobic spore-forming gram-positive rods - Bacillus spp. - what must be ruled out? how?
Must rule out B. anthracis
-often done via PCR
Aerobic spore-forming gram-positive rods - Bacillus anthracis - pathogenicity
-one of the most highly pathogenic microorganisms
-causes anthrax
-contracted from contaminated hides, wool, meat
Aerobic spore-forming gram-positive rods - Bacillus anthracis - gram stain
-large with square ends
-may be in chains
-oval, central to subterminal spores that aren’t swollen
-looks like bamboo
-spores may not be seen in direct smear
Aerobic spore-forming gram-positive rods - Bacillus anthracis - culture
-large
-adherent
-NON-HEMOLYTIC
-flat to slightly convex
-irregular border
-GROUND GLASS appearance
-comma shaped projections
-stands up like BEATEN EGG WHITE when touched with loop
Aerobic spore-forming gram-positive rods - Bacillus anthracis - key characteristics
-NON-HEMOLYTIC (beta-hydrolysis rules out)
-catalase positive
-NON-MOTILE
-capsules are seen in CSF & blood smears
Aerobic spore-forming gram-positive rods - Bacillus anthracis - antibotic susceptibility
Most strains are susceptible to penicillin.
Aerobic spore-forming gram-positive rods - Bacillus cereus - pathogenicity
-often environmental contaminant
-can cause focal & systemic infections, food poisoning
Aerobic spore-forming gram-positive rods - Bacillus cereus - gram stain
-large with square ends
-may be in chains
-oval, central to subterminal spores that aren’t swollen
-looks like bamboo
-spores may not be seen in direct smear
Aerobic spore-forming gram-positive rods - Bacillus cereus - culture
-large
-adherent
-HEMOLYTIC
-flat to slightly convex
-irregular border
-GROUND GLASS appearance
-comma shaped projections
-stands up like BEATEN EGG WHITE when touched with loop
Aerobic spore-forming gram-positive rods - Bacillus cereus - key characteristics
-hemolytic
-motile
Aerobic NON-spore-forming gram-positive rods - Corynebacterium spp. - pathogenicity
-normal on skin & mucous membranes
-opportunistic pathogen
Aerobic NON-spore-forming gram-positive rods - Corynebacterium spp. - gram stain
-irregular
-slightly curved
-non-paralleled sides
-CLUB-SHAPED ends
-if taken from fluid, they can arrange in PALLISADES or clusters with angular V-shaped forms
Aerobic NON-spore-forming gram-positive rods - Corynebacterium spp. - culture
-facultative anaerobes
-grow on most media
Aerobic NON-spore-forming gram-positive rods - Corynebacterium spp. - also known as?
DIPTHEROIDS - because they resemble C. diphtheriae
Aerobic NON-spore-forming gram-positive rods - Corynebacterium spp. - key characteristics
-catalase positive
-non-motile
Aerobic NON-spore-forming gram-positive rods - Corynebacterium diphtheriae - pathogenicity
Diphtheria
Aerobic NON-spore-forming gram-positive rods - Corynebacterium diphtheriae - gram stain
-irregular
-slightly curved
-non-paralleled sides
-CLUB-SHAPED ends
-if taken from fluid, they can arrange in PALLISADES or clusters with angular V-shaped forms
Aerobic NON-spore-forming gram-positive rods - Corynebacterium diphtheriae- culture
-Tindale agar: gray-black colonies with brown halos
-Cystine tellurite agar: black colonies
Aerobic NON-spore-forming gram-positive rods - Corynebacterium diphtheriae - key characteristics
-catalase positive
-non-motile
-toxin producing
Aerobic NON-spore-forming gram-positive rods - Corynebacterium diphtheriae - requirements for ID
-demonstration of toxin production
PCR or MALDI-TOFF are used for ID but must be followed up with ELEK immunoprecipitation test to confrim toxin production
Aerobic NON-spore-forming gram-positive rods - Corynebacterium diphtheriae - Corynebacterium ulcerans
-closely releated to C. diphtheriae
-also contains the TOX gene and can cause diphtheriae
Aerobic NON-spore-forming gram-positive rods - Corynebacterium jeikeium - pathogenicity
hospital-acquired pathogen
Aerobic NON-spore-forming gram-positive rods - Corynebacterium jeikeium - gram stain
-irregular
-slightly curved
-non-paralleled sides
-CLUB-SHAPED ends
-if taken from fluid, they can arrange in PALLISADES or clusters with angular V-shaped forms
Aerobic NON-spore-forming gram-positive rods - Corynebacterium jeikeium - culture
Same as other commensal corynebacteria
Aerobic NON-spore-forming gram-positive rods - Corynebacterium jeikeium - key characteristics
-catalase positive
-rapid sucrose urea (RSU) negative
Aerobic NON-spore-forming gram-positive rods - Corynebacterium jeikeium - antibiotic susceptibility
multidrug resistant (MDR)
Which diphtheroid is the most commonly isolated?
A. C. diphtheriae
B. C. ulcerans
C. C. jeikeium
D. None of the above
C. C. jeikeium
Aerobic NON-spore-forming gram-positive rods - Listeria monocytogenes - pathogenicity
-able to cross the placenta and can infect the fetus causing preterm labor and other complications resulting in fetal loss up to 24%
-meningitis & septicemia in newborns & immunocompromised
-food poisoning with mortality ranges up to 50%
Aerobic NON-spore-forming gram-positive rods - Listeria monocytogenes - gram stain
-parallel sides
-rounded ends
-coccobacillary
-singles or short chain arrangement
Aerobic NON-spore-forming gram-positive rods - Listeria monocytogenes - culture
-tiny colonies with narrow zone of indistinct beta hemolysis
-translucent, gray
Aerobic NON-spore-forming gram-positive rods - Listeria monocytogenes - key characteristics
-catalase positive
-hippurate hydrolysis positive
-esculin positive
-CAMP positive (hemolysis looks like SHOVEL, not arrowhead)
-TUMBLING motility on wet mount
-UMBRELLA growth in motility agar at room temperature but not at 35*C.
Aerobic NON-spore-forming gram-positive rods - Listeria monocytogenes:
-growth temp?
-enrichment?
-differentiation from group B strep?
-differentiation from diphtheroids?
-MALDI-TOF?
-grows from 0.5C-45C
-cold enrichment may be used
-catalase differentiates from group B strep
-motility (tumbling) differentiates from diphtheroids (non-motile)
-MALDI-TOF can be used for ID
Aerobic NON-spore-forming gram-positive rods - Nocardia - pathogenicity
-immunocompetent: skin infection
-immunocompromised: invasive pulmonary & disseminated infections
Aerobic NON-spore-forming gram-positive rods - Nocardia - gram stain
-aerobic actinomycetes
-fine branching filaments with fragmentation
-often beaded
Aerobic NON-spore-forming gram-positive rods - Nocardia - culture
-slow growing
-SBA:
–wrinkled
–dry
–crumbly
–chalky white to orange-tan
–beta hemolytic
Aerobic NON-spore-forming gram-positive rods - Nocardia - key characteristics
-catalase positive
-partially ACID FAST
-SULFUR granules
Aerobic NON-spore-forming gram-positive rods - Nocardia - frequently isolated species
N. brasiliensis
N. pseudobrasiliensis
N. abscessus
Aerobic NON-spore-forming gram-positive rods - Nocardia - reporting
Nocardia isolates may be identified and reported at “complex” or “group” level.
Neisseria & Moraxella - Neisseria spp. - gram stain
-gram negative DIPLOCOCCI (GNDC) OR single COFFEE-BEAN shaped
Neisseria & Moraxella - Neisseria spp. - culture
-aerobic OR facultative
-pathogens are CAPNOPHILIC
Neisseria & Moraxella - Neisseria spp. - key characteristics
-catalase positive
-oxidase positive
Neisseria & Moraxella - Neisseria gonorrhoeae - pathogenicity
-gonorrhea
-salpingitis
-ophthalmia of newborn
Neisseria & Moraxella - Neisseria gonorrhoeae - gram stain
-intracellular & extracellular GNDC
–diagnostic in urethral discharge from symptomatic males
–culture confirmation required for females
Neisseria & Moraxella - Neisseria gonorrhoeae - culture
-requires increased CO2
-usually does NOT grow on SBA
-grows on CHOC & Neisseria-selective media
-colonies are small, grayish white or tan
-may look like mixed culture
Neisseria & Moraxella - Neisseria gonorrhoeae - key characteristics
-oxidase positive
-carbohydrate utilization: GLUCOSE only
Neisseria & Moraxella - Neisseria gonorrhoeae:
-susceptible to what conditions?
-culture used for?
-definitive ID?
-NAATs?
-susceptible to drying & cold
-culture used for presumptive ID
-definitive ID: monoclonal antibodies
-NAATs can be used on male urine specimens with high sensitivity but should NOT be used with oropharyngeal, rectal, ocular, or pediatric patients
Neisseria & Moraxella - Neisseria meningitidis - pathogenicity
-normal flora in upper respiratory tract
-always pathogenic when recovered from usually sterile body fluids such as blood or CSF
-common cause of meningitis in young adults
Neisseria & Moraxella - Neisseria meningitidis - gram stain
-intracellular & extracellular GNDC
-a gram stain of CSF is required for all cases of suspected bacterial meningitis sent to the lab
Neisseria & Moraxella - Neisseria meningitidis - culture
-grows on SBA, CHOC, & Neisseria-selective media
-colonies are grey, convex, glistening, & occasionally mucoid
Neisseria & Moraxella - Neisseria meningitidis - key characteristics
-carbohydrate utilization: GLUCOSE & MALTOSE
-O-nitrophenyl-beta-D-galactopyranoside (ONPG) NEGATIVE
Neisseria & Moraxella - Commensal Neisseria spp. - pathogenicity
-normal in upper respiratory tract
-rarely cause disease
Neisseria & Moraxella - Commensal Neisseria spp. - gram stain
GNDC
Neisseria & Moraxella - Commensal Neisseria spp. - culture
-grow on SBA & CHOC at RT
-grow on nutrient agar at 35*C
-some grow on Neisseria-selective media
-DO NOT require increased CO2
-may be pigmented
Neisseria & Moraxella - Commensal Neisseria spp. - key characteristics
varies with species
Neisseria & Moraxella - Commensal Neisseria spp.:
-speciation?
-must differentiate from?
-N. lactamica?
-not speciated if from respiratory tract
-must differentiate from pathogenic Neisseria when isolated on selective media or from normally sterile body site
-N. lactamica is easily confused with N. meningitidis but N. lactamica acidifies lactose in addition to glucose
Neisseria & Moraxella - Moraxella catarrhalis - pathogenicity
-normal in upper respiratory tract
-causes respiratory infections in young, old, & immunocompromised
-causes otitis media & sinusitis in children
Neisseria & Moraxella - Moraxella catarrhalis - gram stain
GNDC
Neisseria & Moraxella - Moraxella catarrhalis - culture
-grows on SBA & CHOC
-some may grow at RT &/or on Neisseria-selective media
-“HOCKEY PUCK colonies” - colonies can be pushed over agar surface with a loop
Neisseria & Moraxella - Moraxella catarrhalis - key characteristics
-catalase positive
-oxidase positive
-carbohydrate utilization: NEG for all sugars
-ONPG negative
-DNase positive
-butyrate esterase positive
Neisseria & Moraxella - Moraxella catarrhalis - differentiation from Neisseria spp.
DNase & butyrate esterase differentiate from Neisseria spp.
Characteristics of Enterobacterales - gram stain
Non-spore-forming GNRs
Characteristics of Enterobacterales - gram stain
Non-spore-forming GNRs
Characteristics of Enterobacterales - growth characteristics
facultative anaerobes
Characteristics of Enterobacterales - colonies on SBA
most are large, dull, gray, non-hemolytic
Characteristics of Enterobacterales - colonies on MAC
-lactose fermenters = pink
-non-lactose fermenters = colorless
Characteristics of Enterobacterales - Biochemicals
-ferment GLUCOSE
-oxidase negative
-most reduce nitrates to nitrites
-most are catalase positive
List the biochemical tests used for identification of Enterobacterales. (14)
- Oxidase
- nitrate reduction
- carbohydrate fermentation
- ONPG
- H2S production
- Triple sugar iron (TSI) agar
- Indole
- Methyl red (MR)
- Voges-Proskauer (VP)
- Citrate
- Urease
- Phenylalanine deaminase (PD)
- Decarboxylase reactions
- motility
Biochemical tests for identification of Enterobacterales - oxidase - principle
tetramethyl-para-phenylenediamine dihydrochloride reacts with cytochrome C to produce a blue or purple color
Biochemical tests for identification of Enterobacterales - oxidase - interpretation
Positive = blue or purple color
Biochemical tests for identification of Enterobacterales - oxidase:
-test used to differentiate?
-false pos?
-good test to differentiate Enterobacterales from non-fermenters
-false positive from iron-containing wire - use platinum or wooden stick
Biochemical tests for identification of Enterobacterales - nitrate reduction - principle
If organism reduces nitrates to nitrites, a RED color develops when sulfanic aicd & N,N-dimethyl-L-naphthylamine added
Biochemical tests for identification of Enterobacterales - nitrate reduction - interpretation
Positive = red, OR no color after addition of zinc dust
Biochemical tests for identification of Enterobacterales - nitrate reduction - zinc dust
If no color develops (red = pos), zinc dust is added. It reduces nitrates.
-RED color AFTER zinc dust = presence of nitrates (NEG REACTION)
-NO COLOR after zinc dust means nitrates reduces to N2 or NO2 (POS REACTION)
Biochemical tests for identification of Enterobacterales - carbohydrate fermentation - principle
when carbohydrate is fermented, acidic end products cause color change in pH indicator
Biochemical tests for identification of Enterobacterales - carbohydrate fermentation - interpretation
with phenol red indicator = change from red to yellow
Biochemical tests for identification of Enterobacterales - carbohydrate fermentation:
-frequently tested carbohydrates?
-all Enterobacterales ferment?
-frequently tested carbohydrates are glucose, lactose, sucrose, mannose, sorbitol, mannitol, xylose, adonitol, cellobiose, dulcitol, trehalose
-All Enterobacterales ferment GLUCOSE
Biochemical tests for identification of Enterobacterales - ONPG - principle
ONPG is changed to orthonitrophenol by beta-galactosidase
Biochemical tests for identification of Enterobacterales - ONPG - interpretation
positive = yellow
Biochemical tests for identification of Enterobacterales - ONPG:
-test for?
-differentiation of?
-test for slow lactose fermentation
-helpful in differentiating Citrobacter (positive) from most Salmonella (negative).
Biochemical tests for identification of Enterobacterales - H2S production - principle
-organisms that possess H2S-producing enzymes produce colorless H2S gas from sulfur containing compounds
-H2S reacts with iron salt in medium to form black ferrous sulfide
Biochemical tests for identification of Enterobacterales - H2S production - interpretation
black precipitate
Biochemical tests for identification of Enterobacterales - H2S production:
-sulfur-containing compounds?
-differentiates?
-sulfur-containing compounds = sodium thiosulfate, cystine, methionine
-good test to differentiate Salmonella (H2S positive) from Shigella (H2S negative).
Biochemical tests for identification of Enterobacterales - Triple sugar iron (TSI) agar - principle
sugar fermentation produces acid = changes color of pH indicator
Biochemical tests for identification of Enterobacterales - Triple sugar iron (TSI) agar - interpretation
Yellow = acid (A)
Pink = alkaline (K)
Pink butt = glucose NOT fermented
Yellow butt = glucose fermented
Pink slant = lactose/sucrose NOT fermented
Yellow slant = lactose and/or sucrose fermented
Black precipitate = H2S produced
Bubbles = gas production
Biochemical tests for identification of Enterobacterales - Triple sugar iron (TSI) agar:
-contains?
-how to leave cap to tube?
-record as?
-Kligler iron agar (KIA)?
-contains 0.1% glucose, 1% lactose, 1% sucrose, phenol red, sodium thiosulfate, iron salt
-leave cap slightly loose
-record as slant/butt, e.g., K/A [some record NC = no change (for pink butt]
-Kligler iron agar (KIA) is same except NO sucrose
Biochemical tests for identification of Enterobacterales - Indole - principle
-tryptophanase deaminates tryptophan = indole produced
-Kovacs reagent (paradimethylaminobenzaldehyde) forms pink-colored complex with indole
Biochemical tests for identification of Enterobacterales - Indole - interpretation
positive = pink
Biochemical tests for identification of Enterobacterales - Indole - spot indole test
-commercially available
-need source of tryptophan
-use colonies from SBA or CHOC agar, not MAC
-positive = blue
-E. coli & Proteus vulgaris are positive
Biochemical tests for identification of Enterobacterales - methyl red (MR) - principle
-acid products formed when glucose is metabolized by mixed acid fermentation pathway
-color changes pH indicator
Biochemical tests for identification of Enterobacterales - methyl red (MR) - interpretation
positive = red color after addition of MR (pH indicator)
Biochemical tests for identification of Enterobacterales - methyl red (MR) - pH at which test becomes positive?
4.5
Biochemical tests for identification of Enterobacterales - Voges-Proskauer (VP) - principle
acetoin is produced from alternate pathway for glucose metabolism
Biochemical tests for identification of Enterobacterales - Voges-Proskauer (VP) - interpretation
positive = red color when alphanaphthol & KOH added
Biochemical tests for identification of Enterobacterales - Voges-Proskauer (VP) - VP vs. MR
organisms that are VP positive are usually MR negative & vice versa
Biochemical tests for identification of Enterobacterales - Citrate - principle
if organism can use citrate as sole source of carbon, pH increases, and pH indicator changes color
Biochemical tests for identification of Enterobacterales - Citrate - interpretation
positive = green to blue, or growth
Biochemical tests for identification of Enterobacterales - Citrate - false negatives
false negative if cap is NOT loose
Biochemical tests for identification of Enterobacterales - Citrate - positive bacteria
Klebsiella & Enterobacter are positive
Biochemical tests for identification of Enterobacterales - Urease - principle
-urease breaks down to urea
-ammonia released, pH increased, pH indicator changes color
Biochemical tests for identification of Enterobacterales - Urease - interpretation
positive = yellow to pink
Biochemical tests for identification of Enterobacterales - Urease - rapid urease producers
Proteus & Morganella
Biochemical tests for identification of Enterobacterales - Phenylalanine deaminase (PD) - principle
PD deaminates phenylalanine to phenylpyruvic acid, which reacts with ferric chloride to produce a green color.
Biochemical tests for identification of Enterobacterales - Phenylalanine deaminase (PD) - interpretation
positive = green color after addition of ferric chloride
Biochemical tests for identification of Enterobacterales - Phenylalanine deaminase (PD) - positive organisms
-Proteus
-Providencia
-Morganella
Biochemical tests for identification of Enterobacterales - Decarboxylase reactions - principle
if organism has enzyme to decarboxylate amino acid (e.g., ornithine, lysine, arginine), pH increases, pH indicator changes color
-Ornithine decarboxylase (ODC)
Biochemical tests for identification of Enterobacterales - Decarboxylase reactions - interpretation
positive = yellow to purple
Biochemical tests for identification of Enterobacterales - Motility - interpretation
motile organisms grow away from stab line in motility medium
Biochemical tests for identification of Enterobacterales - Motility - interpretation
positive:
-movement away from stab line OR
-hazy appearance throughout medium after overnight incubation
Biochemical tests for identification of Enterobacterales - Motility - positive organisms
most Enterobacterales are positive, EXCEPT Klebsiella & Shigella
Biochemical tests for identification of Enterobacterales - Motility - differentiates?
Differentiates:
-Klebsiella (neg) from Enterobacter (pos)
AND
-Shigella (neg) from Salmonella (pos)
Antigens of Enterobacterales - O antigen:
-alternate name
-location
-characteristics
-used for serological grouping of?
-somatic antigen
-cell wall
-lipopolysaccharide, heat stable
-used for serological grouping of Salmonella & Shigella
Antigens of Enterobacterales - H antigen:
-alternate name
-location
-characteristics
-used to serotype?
-flagellar antigen
-flagella
-proteins, heat labile
-used to serotype Salmonella
Antigens of Enterobacterales - K antigen:
-alternate name
-location
-characteristics
-role in preventing what?
-Vi antigen
-capsular antigen
-capsule
-polysaccharide, heat labile, may mask O antigen; removed by heating
-role in preventing phagocytosis = increased virulence
-Vi antigen is K antigen produced by S. typhi
Escherichia, Shigella, and Salmonella Enterobacterales - E. coli - pathogenicity
-UTI
-septicemia
-neonatal sepsis
-meningitis
-diarrhea (some)
Escherichia, Shigella, and Salmonella Enterobacterales - E. coli - key reactions - POSITIVE
-lactose
-gas
-indole
-MR
-motility
Escherichia, Shigella, and Salmonella Enterobacterales - E. coli - key reactions - NEGATIVE
-H2S
-VP
-citrate
-PD
-urease
Which of the following is the predominant aerobe in the GI tract?
A. Shigella
B. Salmonella
C. E. coli
D. None of the above
C. E. coli
Which of the following is the most common cause of UTI?
A. E. coli
B. Salmonella
C. Shigella
D. None of the above
A. E. coli
Escherichia, Shigella, and Salmonella Enterobacterales - E. coli - EMB media
green metallic sheen
Escherichia, Shigella, and Salmonella Enterobacterales - E. coli - presumptive ID
-lactose positive
-dry colony on MAC
-oxidase negative
-indole positive
Escherichia, Shigella, and Salmonella Enterobacterales - E. coli - E. coli 0157:H7
-does NOT ferment sorbitol
-colorless colonies on SMAC
Escherichia, Shigella, and Salmonella Enterobacterales - Shigella - pathogenicity
-dysentery (shigellosis)
-most communicable of bacterial diarrheas
-found primarily in crowded or substandard conditions, e.g., day-care centers, jails, prisons
Escherichia, Shigella, and Salmonella Enterobacterales - Shigella - key reactions - POSITIVE
MR
Escherichia, Shigella, and Salmonella Enterobacterales - Shigella - key reactions - NEGATIVE
-lactose
-gas
-H2S
-citrate
-PD
-urease
-motility
Escherichia, Shigella, and Salmonella Enterobacterales - Shigella - what is found in the stool of patients positive for Shigella
-blood
-mucus
-neutrophils
Escherichia, Shigella, and Salmonella Enterobacterales - Shigella - most severe species?
S. dysenteriae
Escherichia, Shigella, and Salmonella Enterobacterales - Shigella - most common species in the US?
S. sonnei
Escherichia, Shigella, and Salmonella Enterobacterales - Shigella - serogrouped by what antigens?
O antigens
Escherichia, Shigella, and Salmonella Enterobacterales - Shigella - serogroups?
A, B, C, D
Escherichia, Shigella, and Salmonella Enterobacterales - Shigella - closely related to what organism on a molecular basis?
Escherichia
Escherichia, Shigella, and Salmonella Enterobacterales - Salmonella - pathogenicity
-Typhoid (enteric) fever
-bacteremia
-enterocolitis
Escherichia, Shigella, and Salmonella Enterobacterales - Salmonella - key reactions - POSITIVE
-H2S
-MR
-motility
-lysine decarboxylase (LDC)
Escherichia, Shigella, and Salmonella Enterobacterales - Salmonella - key reactions - NEGATIVE
-lactose
-indole
-VP
-PD
-urease
-ONPG
Escherichia, Shigella, and Salmonella Enterobacterales - Salmonella - what type of meat is it found in?
poultry
Escherichia, Shigella, and Salmonella Enterobacterales - Salmonella - may be transmitted by what type of animal?
reptiles
Escherichia, Shigella, and Salmonella Enterobacterales - Salmonella - S. typhi
-Vi antigen
-only trace H2S
-citrate negative
Escherichia, Shigella, and Salmonella Enterobacterales - Salmonella - grouped by? serotyped by?
-grouped by O antigens (e.g., A, B, C)
-serotyped by H antigens (e.g., 1, 2)
Diarrheagenic Escherichia coli - Enterohemorrhagic (EHEC) - pathogenicity
*also known as Shiga toxin-producing (STEC) OR verotoxin-producing (VTEC)
-diarrhea
-hemorrhagic colitis
-hemolytic uremic syndrome (HUS)
-most common cause of renal failure in children in the US
-may be fatal, especially in young or elderly
Diarrheagenic Escherichia coli - Enterohemorrhagic (EHEC) - transmission
-undercooked meat
-raw milk
-apple cider
Diarrheagenic Escherichia coli - Enterohemorrhagic (EHEC) - disease mechanism
toxins (verotoxins or Shiga toxins)
Diarrheagenic Escherichia coli - Enterohemorrhagic (EHEC) - gram stain of stool
RBCs but usually no segmented neutrophils
Diarrheagenic Escherichia coli - Enterohemorrhagic (EHEC) - E. coli 0157:H7
-most common isolate of group
-pathogen most often isolated from bloody stools
Diarrheagenic Escherichia coli - Enterohemorrhagic (EHEC) - DNA probes
ID genes that code for toxins
Diarrheagenic Escherichia coli - Enterotoxigenic (ETEC) - pathogenicity
-traveler diarrhea
-diarrhea in infants
Diarrheagenic Escherichia coli - Enterotoxigenic (ETEC) - transmission
contaminated food or water
Diarrheagenic Escherichia coli - Enterotoxigenic (ETEC) - disease mechanism
toxins
Diarrheagenic Escherichia coli - Enterotoxigenic (ETEC) - gram stain of stool
no segmented neutrophils or RBCs
Diarrheagenic Escherichia coli - Enterotoxigenic (ETEC) - type of stool
profuse, watery stool
Diarrheagenic Escherichia coli - Enterotoxigenic (ETEC) - DNA probes
detect toxins or toxin genes
Diarrheagenic Escherichia coli - Enteroinvasive (EIEC) - pathogenicity
-bloody diarrhea
-dysentery-like
-usually in young children in areas of poor sanitation
Diarrheagenic Escherichia coli - Enteroinvasive (EIEC) - transmission
contaminated food or water
Diarrheagenic Escherichia coli - Enteroinvasive (EIEC) - disease mechanism
invasiveness
Diarrheagenic Escherichia coli - Enteroinvasive (EIEC) - gram stain of stool
-segmented neutrophils
-RBCs
-mucus
Diarrheagenic Escherichia coli - Enteropathogenic (EPEC) - pathogenicity
-diarrhea in infants
-major pathogen in infants in developing countries
Diarrheagenic Escherichia coli - Enteropathogenic (EPEC) - transmission
formula & food contaminated with fecal material
Diarrheagenic Escherichia coli - Enteropathogenic (EPEC) - disease mechanism
adherence-attachment
Diarrheagenic Escherichia coli - Enteropathogenic (EPEC) - gram stain of stool
no segmented neutrophils or RBCs
Diarrheagenic Escherichia coli - Enteropathogenic (EPEC) - type of stool
watery diarrhea with mucus
Diarrheagenic Escherichia coli - Enteroaggregative (EAEC) - pathogenicity
-diarrhea in developing countries
-chronic diarrhea in HIV-infected patients
Diarrheagenic Escherichia coli - Enteroaggregative (EAEC) - transmission
nosocomial & community acquired
Diarrheagenic Escherichia coli - Enteroaggregative (EAEC) - disease transmission
adherence-attachment
Diarrheagenic Escherichia coli - Diffusely adherent (DAEC) - pathogenicity
-diarrhea & UTI
-most common in children in developing countries
Diarrheagenic Escherichia coli - Diffusely adherent (DAEC) - transmission
little known about epidemiology
Diarrheagenic Escherichia coli - Diffusely adherent (DAEC) - disease mechanism
adherence-attachment
Klebsiella and other commonly isolated Enterobacterales - Klebsiella pneumoniae - pathogenicity
-pneumonia
-UTI
-septicemia
-Carbapenem-resistant
-significant pathogen in both community and hospital settings
Klebsiella and other commonly isolated Enterobacterales - Klebsiella pneumoniae - key reactions - POSITIVE
-lactose
-gas
-VP
-citrate
-urease (slow)
Klebsiella and other commonly isolated Enterobacterales - Klebsiella pneumoniae - key reactions - NEGATIVE
-H2S
-indole
-MR
-PD
-motility
-ODC
Klebsiella and other commonly isolated Enterobacterales - Klebsiella pneumoniae - encapsulated?
yes
Klebsiella and other commonly isolated Enterobacterales - Klebsiella pneumoniae - colonies on agar
colonies are usually mucoid
Klebsiella and other commonly isolated Enterobacterales - Klebsiella pneumoniae - K. oxytoca
similar to K. pneumoniae EXCEPT indole positive
Klebsiella and other commonly isolated Enterobacterales - Klebsiella pneumoniae - differentiation from Enterobacter
-motility
-ODC
Klebsiella and other commonly isolated Enterobacterales - Citrobacter - pathogenicity
-nosocomial infections
-UTI is most common site of infection
Klebsiella and other commonly isolated Enterobacterales - Citrobacter - key reactions - POSITIVE
-gas
-H2S
-MR
-citrate
-motility
-ONPG
Klebsiella and other commonly isolated Enterobacterales - Citrobacter - key reactions - NEGATIVE
-VP
-PD
-LDC
Klebsiella and other commonly isolated Enterobacterales - Citrobacter - lactose?
lactose variable
Klebsiella and other commonly isolated Enterobacterales - Citrobacter - differentiation from Salmonella?
-ONPG
-LDC
Klebsiella and other commonly isolated Enterobacterales - Edwardsiella tarda - pathogenicity
-opportunistic
-can cause bacteremia, wound, and GI infections
-environmental risk factors include exposure to brackish water or undercooked fish
Klebsiella and other commonly isolated Enterobacterales - Edwardsiella tarda - key reactions - POSITIVE
-gas
-H2S
-indole
-MR
-motility
Klebsiella and other commonly isolated Enterobacterales - Edwardsiella tarda - key reactions - NEGATIVE
-lactose
-VP
-citrate
-PD
-urease
Klebsiella and other commonly isolated Enterobacterales - Edwardsiella tarda - primarily what type of pathogen?
fish
Klebsiella and other commonly isolated Enterobacterales - Edwardsiella tarda - differentiation from Salmonella
positive indole
Klebsiella and other commonly isolated Enterobacterales - Klebsiella aerogenes - pathogenicity
-opportunistic & nosocomial infections
-UTI
-respiratory tract infection (RTI)
-wound infections
Klebsiella and other commonly isolated Enterobacterales - Klebsiella aerogenes - key reactions - POSITIVE
-VP
-ONPG
-ODC
-citrate
-motility
-gas
Klebsiella and other commonly isolated Enterobacterales - Klebsiella aerogenes - key reactions - NEGATIVE
-H2S
-indole
-MR
-PD
Klebsiella and other commonly isolated Enterobacterales - Klebsiella aerogenes - colonies on agar
colonies may be mucoid
Klebsiella and other commonly isolated Enterobacterales - Enterobacter cloacae - pathogenicity
-nosocomial infections from contaminated medical devices and instruments
Klebsiella and other commonly isolated Enterobacterales - Enterobacter cloacae - key reactions - POSITIVE
-ODC
-VP
-sucrose
Klebsiella and other commonly isolated Enterobacterales - Enterobacter cloacae - key reactions - NEGATIVE
-oxidase
-H2S
-indole
Klebsiella and other commonly isolated Enterobacterales - Enterobacter cloacae - antibiotic susceptibility
frequently antibiotic resistant (intrinsic and plasma-based)
Other commonly isolated Enterobacterales - Serratia marcescens - pathogenicity
-opportunistic pathogen
-UTI (indwelling catheters)
-ocular
-GI tract in children
Other commonly isolated Enterobacterales - Serratia marcescens - key reactions - POSITIVE
-VP
-citrate
-motility
Other commonly isolated Enterobacterales - Serratia marcescens - key reactions - NEGATIVE
-lactose
-H2S
-indole
-PD
-urease
Other commonly isolated Enterobacterales - Serratia marcescens - pigment produced when incubated at RT
red pigment
Other commonly isolated Enterobacterales - Proteus vulgaris & Proteus mirabilis - pathogenicity
-UTI
-wound infections
-septicemia
Other commonly isolated Enterobacterales - Proteus vulgaris & Proteus mirabilis - key reactions - POSITIVE
-H2S
-MR
-PD
-urease
-motility
Other commonly isolated Enterobacterales - Proteus vulgaris & Proteus mirabilis - key reactions - NEGATIVE
-lactose
Other commonly isolated Enterobacterales - Proteus vulgaris & Proteus mirabilis - colonies on agar
-swarming
-burned chocolate odor
Other commonly isolated Enterobacterales - Proteus vulgaris & Proteus mirabilis - indole
P. mirabilis - indole negative
P. vulgaris - indole positive
Other commonly isolated Enterobacterales - Proteus vulgaris & Proteus mirabilis - P. vulgaris on TSI
A/A - because of sucrose fermentation
Other commonly isolated Enterobacterales - Morganella morganii - pathogenicity
-mainly nosocomial infections
-UTI
-wound infections
Other commonly isolated Enterobacterales - Morganella morganii - key reactions - POSITIVE
-indole
-MR
-PD
-urease (weak)
-motility
-ODC
Other commonly isolated Enterobacterales - Morganella morganii - key reactions - NEGATIVE
-lactose
-H2S
-VP
-citrate
Other commonly isolated Enterobacterales - Providencia - pathogenicity
-UTI
-diarrhea
Other commonly isolated Enterobacterales - Providencia - key reactions - POSITIVE
-indole
-MR
-citrate
-PD
-motility
Other commonly isolated Enterobacterales - Providencia - key reactions - NEGATIVE
-lactose
-H2S
-VP
Other commonly isolated Enterobacterales - Providencia - urease
P. rettgeri is urease positive
Other commonly isolated Enterobacterales - Providencia - which species is most commonly isolated?
P. stuartii
Other commonly isolated Enterobacterales - Yersinia enterocolitica - pathogenicity
diarrhea
Other commonly isolated Enterobacterales - Yersinia enterocolitica - key reactions - POSITIVE
-MR
-urease
-ONDC
Other commonly isolated Enterobacterales - Yersinia enterocolitica - key reactions - NEGATIVE
-lactose
-H2S
-VP (variable)
-citrate
-PD
Other commonly isolated Enterobacterales - Yersinia enterocolitica - gram stain
-gram negative coccobacilli
-bipolar staining
Other commonly isolated Enterobacterales - Yersinia enterocolitica - optimal temp for growth
25-30*C
Other commonly isolated Enterobacterales - Yersinia enterocolitica - motility
motile at 25C but NOT 35C
Other commonly isolated Enterobacterales - Yersinia enterocolitica - agar
-CIN agar is selective
-incubate 48 hours
-red “BULL’S EYE” colonies surrounded by colorless halo
What species of Yersinia causes plague?
Y. pestis
Summary of key reactions for Enterobacterales - lactose negative
-Shigella
-Edwardsiella
-Salmonella
-Citrobacter (some)
-Serratia
-Proteus
-Morganella
-Providencia
-Yersinia
Summary of key reactions for Enterobacterales - H2S positive
-Edwardsiella
-Salmonella
-Citrobacter
-Proteus
Summary of key reactions for Enterobacterales - VP positive
-Klebsiella
-Enterobacter
-Serratia
Summary of key reactions for Enterobacterales - PD positive
-Proteus
-Morganella
-Providencia
-Klebsiella (slow)
Summary of key reactions for Enterobacterales - Urease positive
-Proteus
-Morganella
-Providencia rettgeri
-some species of Citrobacter
Summary of key reactions for Enterobacterales - nonmotile at 35*C
-Shigella
-Klebsiella
-Yersinia (motile at 22*C)
Appearance of Enterobacterales on selected media - Escherichia coli - TSI
A/A, gas
Appearance of Enterobacterales on selected media - Escherichia coli - MAC
flat, dry pink colony with darker pink halo
Appearance of Enterobacterales on selected media - Escherichia coli - HE
yellow
Appearance of Enterobacterales on selected media - Escherichia coli - XLD
yellow
Appearance of Enterobacterales on selected media - Shigella - TSI
K/A
Appearance of Enterobacterales on selected media - Shigella - MAC
colorless
Appearance of Enterobacterales on selected media - Shigella - HE
green
Appearance of Enterobacterales on selected media - Shigella - XLD
colorless
Appearance of Enterobacterales on selected media - Edwardsiella - TSI
K/A, gas, H2S
Appearance of Enterobacterales on selected media - Edwardsiella - MAC
colorless
Appearance of Enterobacterales on selected media - Edwardsiella - HE
colorless
Appearance of Enterobacterales on selected media - Edwardsiella - XLD
red, yellow, or colorless with or without black centers
Appearance of Enterobacterales on selected media - Citrobacter - TSI
A/A or K/A, gas, with or without H2S
Appearance of Enterobacterales on selected media - Citrobacter - MAC
-colorless at 24 hrs
-may become pink at 48 hrs
Appearance of Enterobacterales on selected media - Citrobacter - HE
colorless
Appearance of Enterobacterales on selected media - Citrobacter - XLD
red, yellow, or colorless with or without black centers
Appearance of Enterobacterales on selected media - Salmonella - TSI
K/A, gas, H2S
Appearance of Enterobacterales on selected media - Salmonella - MAC
colorless
Appearance of Enterobacterales on selected media - Salmonella - HE
green
Appearance of Enterobacterales on selected media - Salmonella - XLD
red with black center
Appearance of Enterobacterales on selected media - Klebsiella - TSI
A/A, gas
Appearance of Enterobacterales on selected media - Klebsiella - MAC
pink, mucoid
Appearance of Enterobacterales on selected media - Klebsiella - HE
yellow
Appearance of Enterobacterales on selected media - Klebsiella - XLD
yellow
Appearance of Enterobacterales on selected media - Enterobacter - TSI
A/A, gas
Appearance of Enterobacterales on selected media - Enterobacter - MAC
pink, may be mucoid
Appearance of Enterobacterales on selected media - Enterobacter - HE
yellow
Appearance of Enterobacterales on selected media - Enterobacter - XLD
yellow
Appearance of Enterobacterales on selected media - Serratia - TSI
K/A
Appearance of Enterobacterales on selected media - Serratia - MAC
-colorless at first, turning pink
-S. marcescens may have red pigment at RT
Appearance of Enterobacterales on selected media - Serratia - HE
colorless
Appearance of Enterobacterales on selected media - Serratia - XLD
yellow or colorless
Appearance of Enterobacterales on selected media - Proteus - TSI
-K/A (P. mirabilis)
-A/A (P. vulgaris)
-gas, H2S
Appearance of Enterobacterales on selected media - Proteus - MAC
colorless, may swarm
Appearance of Enterobacterales on selected media - Proteus - HE
colorless
Appearance of Enterobacterales on selected media - Proteus - XLD
yellow or colorless, with or without black centers
Appearance of Enterobacterales on selected media - Morganella - TSI
K/A, gas
Appearance of Enterobacterales on selected media - Morganella - MAC
colorless
Appearance of Enterobacterales on selected media - Morganella - HE
colorless
Appearance of Enterobacterales on selected media - Morganella - XLD
red or colorless
Appearance of Enterobacterales on selected media - Providencia - TSI
K/A
Appearance of Enterobacterales on selected media - Providencia - MAC
colorless
Appearance of Enterobacterales on selected media - Providencia - HE
colorless
Appearance of Enterobacterales on selected media - Providencia - XLD
yellow or colorless
Appearance of Enterobacterales on selected media - Yersinia - TSI
Yellow/orange