ENTEROBACTERIACEAE MUG positive (methyl umbelliferyl beta D-glucoronide
except E. coli 0157:H7)
General characteristics: Sorbitol MAC (+) except E. coli 0157:H7
- All are gram negative bacilli #1 cause of UTI because of virulence factor which is
- All ferments glucose PILI – for adherence, capable of attaching itself to
- All are motile except Klebsiella, Shigella and Yersinia Urinary tract
o Non motile at 22C (room temp.) – Shigella, #1 gram negative that causes sepsis
Klebsiella, Yersinia pestis #2 cause of neonatal meningitis (#1 Streptococcus
- All are catalase Positive except Shigella dysenteriae agalactiae)
- All reduce Nitrate to nitrite Nosocomial, wound, bacteremia, pneumonia
- All are oxidase negative except for Plesiomonas
- Selective media for Enterobacteriaceae: BIOTYPE: ESCHERICHIA COLI INFECTIONS:
- MAC A. Enterotoxigenic E.coli (ETEC)
- EMB - traveller’s diarrhea (disease)
- DCA - watery diarrhea
- SSA - Cholera like toxin (heat labile enterotoxin)
- BSA - O6;O8;O25
- Virulence factor: B. Enteropathogenic E.coli (EPEC)
- Adhere in tissue - Infantile diarrhea (pathogenicity island)
- Colonize a part of body - Stool without blood
- Produce toxin - gastrointestinal diarrhea
- Invade tissues - O111,O114
- Antigens (serological testing): C. Enteroinvasive E.coli (EIEC)
- Capsule (K antigen) – heat labile - Dysentery (shigella) like diarrhea (invasion)
- Cell wall (O or somatic antigen) – heat stable - Blood and mucous in feces
- Flagella (H antigen) – heat labile - Stool with WBCs
- Somatic antigen is heat stable - Virulence factor: invasin
- Virulence antigen (K and H) is heat labile - O124,143,164
- Salmonella typhi both possesses somatic and virulence - Sereny test - virulence test
antigen H antigen o (+) keratoconjuctivitis of rat
- LACTOSE fermenters and Oxidase POSITIVE: D. Enterohemorrhagic E. coli (EHEC)
- Plesiomonas, Vibrio vulnificus, Aeromonas - Verotoxin E. coli (VTEC)
LACTOSE FERMENTERS NON LACTOSE - Hemolytic Uremic Syndrome (HUS), hemorrhagic
FERMENTERS colitis/diarrhea
Most are non-significant Most significant pathogens - Common among adults
No need for susceptibility test Need to do susceptibility - Undercooked hamburger
testing - Shigella like toxin – verotoxin
Pink colony in Mac Conkey Colorless colony - E.coli O157:H7
a) Escherichia coli a) P-M-P (Proteus, - Sorbitol Mac Conckey (-)
b) Klebsiella Morganella, - MUG (-)
c) Enterobacter Providencia)- PAD
E. Enteroaggregative E. coli - acute and chronic diarrhea
d) Serratia (slow LF) positive
e) Hafnia b) Salmonella (aggregative adhesion fimbriae)
f) Yersinia c) Shigella F. Diffusely adherent E. coli (DAEC)
g) Citrobacter d) Edwardsiella - Adhere to the EC in a diffused patterns
h) Arizona - Diarrheal syndrome with watery diarrhea
i) Salmonella arizonae - UTI and diarrhea
j) Shigella sonnei
k) Plesiomonas
2. Enterobacter
MAC: pink colony
1. Escherichia coli (colon bacillus)
TSI: A/A
Primary marker of fecal contamination
IMViC: -,-,+,+
TSI: A/A
Urease NEGATIVE except E. gergoviae
LIA: K/K
Causes UTI, wound infection and septicemia
LOA: +,+,-
IMViC: ++-- LOA REACTION OF ENTEROBACTER:
UREASE NEGATIVE Lysine Ornithine Arginine
Greenish metallic sheen on EMB
E. aerogenes + + - C. koseri/ C. diversus - A/A + +
E. gergoviae + + - Mistaken as E. coli
Hafnia alvei + + - C. amalonaticus A/A + -
E. cloacae - + +
E. sakazakii - + + Urease Lysine Decarboxylase
E. Pantoea - - - Citrobacter + -
agglomerans
Salmonella - +
3. Klebsiella pneumonia - LDC: To differentiate E.coli (+) from C.koseri(-)
Capsulated, nonmotile
TSI: A/A with gas 6. PMP Group (Proteeae)- NLF
LIA: K/K (purple/purple) - Phenylalanine deaminase (PAD) (+)
LOA: +,-,- - Lysine Deamination (+)
IMVic: -,-,+,+ - LIA= R/A (red/yellow)
UREASE AND MALONATE (+) - UREASE (+) except Providencia alcalifacien
MAC - mucoid - LOA -,-,-, except Morganella and Proteus mirabilis
Lactose fermenters (ornithine positive)
Causes Pneumonia, wound, meningitis, UTI, - Indole positive except Proteus mirabilis
ESBL (+) (Extended Spectrum Beta-lactamase) - Proteus
E.coli, Proteus mirabilis, Klebsiella pneumoniae- Swarm on BAP (Motile)
produce ESBL Which is not inhibited by antibiotics
TSI- K/A H2S
Lysine V-P and Indole
decarboxylase Urease LIA- R/A
#2 UTI, renal stone
- Proteus vulgaris - indole (+)
K. pneumoniae + + -
- Proteus mirabilis - indole (-), cause renal stone
K. oxytoca + + + - Proteus vulgaris - associated to
K. ozaenae + - - - OX2
K.rhinoscleromatis - - - - OX19
- K. oxytoca - often mistaken as E. coli but E.coli has no - OXK- P. Mirabilis
capsule TSI Urease Ornithine
- Indole positive – K. Oxytoca Morganella morgani K/A + +
4. Arizona spp. Providencia stuartii K/A +/- -
LLF Late Lactose Fermenter Providencia rettgeri K/A + -
Related to Salmonella
TSI- A/A +H2S (Black precipitate) Providencia alcalifaciens K/A - -
ONPG (+) Proteus mirabilis K/A + + +
LIA- K/K (purple/purple) H2S
S. arizonae - only Salmonella that is lactose fermenter
5. Citrobacter (citrate +) 7. Salmonella (Non Lactose Fermenter)
- LLF - Pathogen of stool
- TSI- A/A - Black colony on SSA (Salmonella-Shigella agar)
- H2S (+) - BSA(Bismuth sulfide agar) - carrier- Black with
- LIA-K/A metallic silver sheen
- ONPG (+) - Wilson-Blair Medium
- Related to Salmonella (LDC +) - TSI: K/A HH2S with gas and H2S production
- Citrobacter diversus- nursery outbreak, neonatal - LIA: K/K
meningitis - Aerogenic except Salmonella typhi and Salmonella
- Citrobacter freundii - UTI, pneumonia, endocarditis, gallinarum
abscess - Motile except S. gallinarum , S. pullorum
- Antigen
TSI Indole Malonate o Vi- virulence
C. freundii A/A H2S - + o O- Cell wall
Cross react with S. typhi o H- flagellar
- O group D antisera (+); S. typhi - Diarrhea, wound infection, bacteremia
- Salmonella paratyphi A= H2S and LDC negative
- Related to Citrobacter 11. Yersinia (LLF)
- Also pathogenic in stool (true pathogen in stool)
Species: A. Yersinia pestis (plague bacillus)
- S. typhi= typhoid fever, meningitis, osteomyelitis - Spread from animal host to man by arthropod vector
- S.paratyphi A and B= paratyphoid fever - Bioterrorism agent
- S. paratyphi C (S. cholera suis)= septicemia - Spread via INHALATION
- Salmonella enteritidis= gastroenteritis - Bipolar stain (Wayson), safety pin appearance
- Other Salmonella species= food poisoning - stalactite (+)
- Treatment: CHLORAMPHENICOL - V and W antigens
- Urease (-)
8. Shigella spp (NLF)= acetate (-) - LOA: -,-,-
- Non motile - Rat flea bite, buboes, bubonic, pneumonic, septicimic
- Colorless on SSA plague, BLACK DEATH
- acetate (-) - Vector: Xenopxylla cheopsis
- TSI: K/A B. Yersinia enterocolitica
- LIA:K/A - oxidase (-)
- LOA -,-,- except S. sonnei (ornithine (+)) - Diarrheal agent
- Related to E.coli (acetate +) - Motile at 22C but not at 35C
- Intestinal pathogen (dysentery)- blood with mucus - TSI: A/A
- Diagnosis - culture of fresh stool with mucous flecks - ONPG (+)
and rectal swab of ulcer (BEST) - Ornithine (+)
- EMB and SS - Urease (+)
- Used for searching cases of Shigellosis - Cold enrichment at 4C (cycrophilic??)
- Bull’s eye colony on CIA (Cefsulodin Irgasan
O Ag Mannito Ornithine ONPG Novobiocin CIN agar)
l
- Aeromonas - CIN (+) but oxidase (+)
S. dysenteriae A - -
- Zoonotic, unpasteurized milk
S. flexneri B + - - Causes enterocolitis, appendicitis
S. boydii C + - C. Yersinia pseudotuberculosis
S. sonnei D + + - animal pathogen
- LOA -,-,-
Differential test for Salmonella and Shigella - UREASE (+)
Salmonella Shigella - Causes Mesenteric Lymphadenitis, septicemia
Motility + -
Y. pestis Y. enterocolitica Y. pseudotb
H2S + -
Motility - + +
LOA +,+,+ -,-,-
Urease - + +
LIA K/K (LDC+) K/A (LDC-)
Ornithine - + -
Indole - +
Invasive - +
WEEK 10: VIBRIONACEAE
Blood culture + -
Related Citrobacter E. coli - Vibrio, Aeromonas, Plesiomonas
- Cause diarrheal diseases but not member of
9. Serratia marcescens (NLF) enterobacteriacae
- Red Pigment (Prodigiosin) - All are OXIDASE (+)
- LOA= +,+,- - All are CATALASE (+)
- LIA= K/K - All are INDOLE (+)
- DNAse, Lipase, Gelatinase, drug resistant - All ferment glucose
- Nosocomial (UTI, Bacteremia, Pneumonia) - All have polar flagella
- TSI A/A or K/A +/-gas (don’t ferment lactose) - To differentiate them with Enterobactericeae- oxidase (-)
- Aeromonas hyrophila
10. Edwardsiella tarda (similar to Salmonella) o Present in fresh water
- TSI= K/A +gas + H2S (Salmonella) o Growth on MAC
- IMViC= +,+,-,- E. coli- lactose fermenters
o B-hemolysis
- Lysine decarboxylase (+)
A. Vibrio - #1 in japan
- curved, comma, Motile - Kanagawa phenomenon - production of betahemolysin
- Produce tetrodotoxin (urotoxin 1mg can cause death of - Halophilic (8% NaCl)
fully grown adult) - Indole (+)
- Oxidase (+) except V. mitschnikovii - Non sucrose fermenters (Green on TCBS)
- 0129 susceptible (antibiotic disc) - Kanagawa (+)
- Halophilic except: V. Cholera, V. mimicus - LOA +,+,-
(Alkalinophilic) - Gastroenteritis (seafoods)
- LOA +,+,-
- Nitrate reduction (+) Disease 8% TCBS
1. Vibrio cholera NaCl
- pathogenic in stool V. cholerae Cholera - Yellow
- VCO-1- Cause pandemic V. alginolyticus gastroenteritis + Yellow
- Nonhalophilic 1-3% NaCl V. parahemolyticus gastroenteritis + Green
- Indole (+)
- Rice watery stool (cholera) - Greater than 10 V. vulnificus Septicemia, + Green
defecation in a day wound Only
- String test positive - 0.5% Na desoxycholate lactose
- Sucrose fermenters (yellow on TCBS) (+)
- Cholera toxin action - cyclic adenosine monophosphate Disease: all cause diarrhea, wound , septicemia
- Heat labile enterotoxin Marine Fresh water Fresh water
- TCBS agar – isolate and differentiate vibrio water
- Indicator: Bromthymol Blue Vibrio- Aeromonas Plesiomonas
Grow in shigelloides-
Biotype Classical El Tor (masama) Brackish Cross
(mabait) water agglutination
Polymixin S R NaCl + - -
Susceptibility
Lysis by + - Motility + + +
bacteriophage Oxidase + + +
Chicken RBC - + 0129 S R S
agglutination
LOA +,+,- +,-,+ +,+,+
Hemolysis of - +
sheep RBC DNase, - + -
Vogues-Proskauer - + Beta BEM Inositol (+)
test TSI A/A,K/A A/A gas,H2S A/A or K/A
Serotype Ogawa Inaba Hikojima
Anti-ogawa + - +
Anti-inaba - + + CAMPYLOBACTER
- curved, S - shaped, seagull wing
- darting motility
Laboratory Diagnosis of V. cholera O1
- zoonotic
1. Darkfield microscopy
- Oxidase +
- Exhibit: darting motility
- Catalase +
- Curve, comma shaped
- Growth at 42C (C. jejuni and C. coli)
2. Culture
- Urease (-)
a. TCBS - Thiosulfate citrate bile salt sucrose
- Indoxyl acetate (+)
b. TTGA - Tellurite taurochocolate gelatin agar
- Associated with animal abortion especially C. fetus
c. APW - alkaline peptone water
- C. jejuni - #1 gastroenteritis in US
3. Oxidase test - positive
- Guillain-Barre Syndrome – target neurons, nerve cells
4. 0129 sensitivity test (S)
5. Polymixin B susceptibility test
37C 42C Nalidixic Cepha- Hippurate
6. Cholera red test = nitrate and indole (+) red acid lothin
C. jejuni + + S R +
2. Vibrio parahemolyticus/ Summer diarrhea
- B-hemolysis on WAGATSUMA AGAR C. coli + + S R -
C. fetus + - R S - - Motile: Monotrichous
- O/F= +/- (yellow/green)
- Grape like odor (Aminoacetaphenone)
Differentiation of Campylobacter and Helicobacter - Growth at 42C
- Cetrimide (+) – selective agar for Pseudomonas
Oxidase, Urease Media Disease - Mac (+)
Catalase, - Nitrate reduction positive
Microaerophile - Acetamide (+)
C. + - Butzler Diarrhea - produce PYOCYANIN (best)
jejuni Skirroe Abortion
CBAP
- pyorubin, pyomelanin, pyoverdin – pseudosel agar
- Fluorescein (P. putida, P. fluorescens)
H. + + Urea agar Peptic ulcer,
pylori gastritis, - LOA- -,-,+
gastrointestinal - Nosocomial infection, #2 most common infection in
carcinoma burn victims, wound, ecthyma gangrenosum, swimmer’s
TYBE B gastritis
Due to chemical ear, contact lens infection, #1 cystic fibrosis,
irritants and stress dermatitis(Jacuzzi)/richman’s disease- earthy dirt
Neutralize acididty like odor
if stomach because - No. 1 nosocomial infection= S. aureus
of urease, shield
with ammonia - Pseudomonas fluorescens
- H. pylori - Scirose medium??, buttsler and CBAP, RIA - produce fluorescein but not pyocyanin
- RIA- Urea breath test, - No growth at 42C
- Synthelation counter- for breath test - Bacteremia, wound, UTI
- Use O-F tube and add Mineral oil - to prevent the entry 2. Burkholderia cepacia
of O2. - oxidase (+)
- Motile(lopotrichous)
Diagnostic for NFO - LDC (+)
- Fermentative - close (fermenters) O-F- +/+ - Pink colonies on mac- lactose oxidizer
- Oxidative- open (oxidizers)- O-F- +/- - Yellow on OFPBL ( oxidative fermentative polymixin B
- Non utilizers- OF -/- Basitracin lactose) or PC agar
- Medium: Hugh and Leifson medium - #2 cystic fibrosis, septicemia, pneumonia
- 1% glucose, 1 % agar; low peptone - Drug resistant
- Indicator: bromthymol blue - Earthy dirt like odor
o (+) yellow- acid
o (-) green (no acid) 3. Burkholderia pseudomallei
- Growth at 42C - agent for bioterrorism
- (+) growth at 35 C and 42C - Pseudomonas aeruginosa - Meliodosis or Glanders like: vietnamese time bomb
- (-) no growth at 42C - P. fluorescens - found in soil
- Cetrimide test- 35C for 7 days - wrinkled colony
o (+) growth- P. aeruginosa - Growth at 42 C
o (-) growth- E. coli - Motile(lopotrichous)
- Litmus milk test (35C for 7 days) - OF- +/- (lactose oxidizer)
o Acid- pink (E. faecium)= coagulate casein 4. Burkholderia mallei
o Hydrolyze casein- straw colored milk - Only non-motile pseudomonad
o Alkaline= blue clot (Alcaligenes faecalis) - Glander’s disease (horse)
- Of- +/- (glucose, lactose, maltose)
o Peptonization- Bulkholderia cepacia
5. Pseudomonas stutzeri
- Oxidase + except S. maltophilia
- non lactose oxidizer
- Motile except B. mallei
- Brown (Buff) colored
- Mac Conkey (+) colorless
- Wrinkled colony
- TSI= K/K
- 6.5% NaCl (+)
- O-F = Yellow (O) Green (F)
- NO2-N2 nitrite reducing
- Oxidize glucose but does not ferment
6. Stenotrophomonas maltophilia
- Opportunistic infection (environment)
- Maltose loving
Species: - Oxidase (-)
1. Pseudomonas aeruginosa - Dnase (+)
- Oxidase + - Indole (+)
- O-F - +/- (Glucose, Maltose) Oxidase Catalase MAC
- Motile (lophotrichous) Acinetobacter - + +
- Lavender green colony Alkaligenes + + +
- Associated with UTI
Flavobacterium + + +/-
- Iatrogenic infection
Moraxella + + -
7. Shewanella putrefaciens
- TSI= K/K H2S Kingella + - -
- Oxidase positive Eikenella + - -
Other Non-Fermentative Oxidizers:
1. Acinetobacter
- Oxidase (-)
- Catalase (+)
- Non motile
- Growth on MAC
a. Acinetobacter anitratus/baumannii (oxidizer)
- Herella vaginocola
b. Acinetobacter lwoffi (non oxidizer)
- Mina polymorpha
- Mistaken as Neisseria (gram – CB)
- UTI, wound, Diarrhea, Resistant to AMA
2. Alkaligenes faecalis
- Found in stool
- Motile - peritrichous
- Assaccharolytic= OF test -/-
- Apple like “fruity “odor
- UTI, Wound and diarrhea
3. Moraxella lacunata (Hockey pock colony)
- Not grown in Mac
- Cuase Blepharoconjuctivitis
- Oxidase (+)
- Catalase (+)
- Assacharolytic
- Mistaken as Neisseria
4. Chryseobacterium (Flavobacterium) meningosepticum
- Associated with immunocompromise patient
- Isolated from respiratory therapy equipmeny
- Produce Flavin (yellow pigment)
- Oxidase (+)
- DNase (+)
- Gelatin HOH (+)
- Indole (+)
- Non motile
- Neonatal meningitis, bacteremia, endocarditis
5. Eikenella corrodens
- found in oral cavity
- Human bite wound
- Corrode agar
- Bleach like odor
6. Kingella kingae
- Cause subacute bacterial endocarditis
- Pits the agar
- No growth in MAC
