MODULE 10: ENTEROBACTERIACEAE Enterobacter Aerogenes

Cloacae
GENERAL CHARACTERISTICS Gergoviae
Cancerogenus
• Enterics (taylorae)
- All ferment glucose Hormaechei
- All reduce nitrate to nitrites
Pantoea Agglomerans
- All are oxidase negative Cronobacter Sakazakii
◼ Except Plesiomonas Hafnia Alvei
• Motility serratia Marcescens
- All motile at body temperature except Liquefasciens
◼ Klebsiella Rubidaea
Fonticola
◼ Shigella Odorifera
◼ Yersinia Plymuthica
Proteeae Proteus Mirabilis
KEY CHARACTERISTICS OF THE FAMILY
Vulgaris
ENTERBACTERIACAEA Penneri
Hauseri
TRIBE GENUS SPECIES
Myxofaciens
Escherichieae Esherichia coli
albertii
Morganella Morganii
blattae
Providencia Alcalifaciens
vulneris
Rettgeri
fergusonii
Stuartii
hermanii
Yersinieae Yersinia Pseudotuberculosis
Pestis
Shigella dysenteriae
Enterolitica
flexneri
Frederiksenii
boydii
Kristensenii
sonnei
Intermedia
Edwarsiellaeae Edwardsiella tarda
Ruckeri
liquecfaciens
hoshinae
ictaluri COLONY MORPHOLOGY
Salmonelleae Salmonella enterica
bongori • Gram-negative coccobacilli or straight rods
Citrobateriaceae Citrobacter freundii - Not very useful in identification other than
koseri (C. diversus)
amalonaticus
ruling out other organisms
youngae • Facultatively anaerobic
braakii - Thioglycolate broth: growth throughout the
farmeri tube
Klebsielleae Klebsiella Pneumoniae • Macroscopic morphology
subsp.
- Large moist, gray colonies
Pneumoniae
- Some mucoid
pneumoniae ◼ Klebsiella
subsp. ozaenae

pneumoniae
subsp.
rhinoscleromatis
varicola
ornitholytica
 GROWTH ON MEDIA BIOCHEMICAL CHARACTERISTICS OF TRIBES OF
ENTEROBACTERIACEAE
• MacConkey (MAC) agar
- Selective and differential ESCHERICHAEAE
- Bile salts and crystal violet inhibit gram H2S (TSI agar) -
positive Urease -
- Lactose fermentation is differential Indole + or -
• Eosin-methylene blue (EMB) agar Methyl red +
- Selective and differential Voges-Proskauer -
- Methylene blue inhibits gram positive Citrate (Simmons) -
- Lactose and sucrose fermentation is KCN -
differential Phenylalanine deaminase -
Mucate d
• Hektoen enteric (HE) agar
Mannitol + or -
- Selective and differential
- Bile salts inhibit gram-positive, some gram-
negative EDWARSIELLEAE
- Lactose and sucrose fermentation is H2S (TSI agar) +
differential Urease -
◼ Most nonpathogens ferment lactose Indole +
and sucrose show orange color (low pH) Methyl red +
◼ Pathogens (salmonella and shigella) Voges-Proskauer -
green to blue green color with H2S gas Citrate (Simmons) -
creating a black precipitation KCN -
• Xylose lysine deoxycholate (XLD) agar Phenylalanine deaminase -
- Selective (less so than MAC and HE) and Mucate -
differential Mannitol -
- Sodium desoxycholate
◼ Inhibits gram-positive, some gram- CITROBACTERICEAE
negative H2S (TSI agar) + or -
- Three carbohydrates, sucrose and lactose in Urease (+w) or −
excess, and xylose with a phenol red Indole − or +
indicator Methyl red +
- Lysine present to detect lysine Voges-Proskauer -
decarboxylation Citrate (Simmons) +
- Thiosulfate present to detect hydrogen KCN + or −
sulfide (H2S) Phenylalanine deaminase -
• XLD agar Mucate
- Yellow colonies Mannitol +
◼ Fermenters or those not producing
lysine decarboxylase Escherichia coli, SALMONELLEAEA
Citrobacter H2S (TSI agar) +
- Colorless or Red colonies Urease -
◼ Shigella Indole -
- Red colonies with black centers Methyl red +
◼ Initially yellow then revert to red Voges-Proskauer -
➢ When lysine is decarboxylated, Citrate (Simmons) +
causing alkaline pH KCN -
>> Salmonella Phenylalanine deaminase -
Mucate d
Mannitol +
 KLEBSIELLEAE - K antigen
H2S (TSI agar) - ◼ Capsular antigen; heat stable
Urease - or (+) ◼ K1 antigen of E.coli, Vi antigen of
Indole - Salmonella typhi
Methyl red -
Voges-Proskauer + CLINICAL SIGNIFICANCE
Citrate (Simmons) + • Two major types
KCN + - Opportunistic pathogens
Phenylalanine deaminase -
◼ Normal flora
Mucate + or -
◼ Infections in other “non-normal” sites
Mannitol +
>> Septicemia, wounds, urinary tract
infections (UTI’s), meningitis
PROTEEAE >> E.coli
H2S (TSI agar) + or - - Primary pathogens
Urease + or - ◼ Salmonella spp.
Indole + or - ◼ Shigella spp.
Methyl red + ◼ Yersinia spp.
Voges-Proskauer - ➢ generally from ingesting
Citrate (Simmons) d contaminated food and water
KCN +
Phenylalanine deaminase + BACTERIAL SPECIES AND THE INFECTIONS THEY
Mucate - COMMONLY PRODUCE
Mannitol - or +
BACTERIAL SPECIES DISEASES
Escherichia coli Bacteriuria
YERSINIAE Septicemia
H2S (TSI agar) - Neonatal sespsis
Urease + Meningitis
Indole + or - Diarrheal syndrome
Methyl red +
Voges-Proskauer - BACTERIAL SPECIES DISEASES
Citrate (Simmons) - Shigella spp. Diarrhea
KCN - Dysentery
Phenylalanine deaminase -
Mucate Edwardsiella spp. Diarrhea
Mannitol + Wound infection
Septicemia
Enteric fever
VIRULENCE AND ANTIGENIC FACTORS

• Adherence: prevent being removed from tissue

BACTERIAL SPECIES DISEASES
• Toxins: food poisoning
Salmonella spp. Septicemia
• Invasive enzymes: spreading factors Enteric fever
• Antibiotic resistance Diarrhea
- Extended spectrum β-Lactamases (ESBL)
• Serology Citrobacter spp. Opportunistic and
- O antigen (somatic antigen) hospital-acquired
◼ Heat stable antigen located in the cell infections (wound,
wall (LPS) urinary)
- H antigen
◼ Flagellar antigen; heat stable
 BACTERIAL SPECIES DISEASES E.coli
Klebsiella spp. Bacteriuria
Pneumonia • Dry pink colonies on MAC plates
Septicemia • Β-hemolysis on blood agar plate (BAP)
• Motile
Enterobacter spp. Opportunistic and • Sex pili, fimbriae
hospital-acquired • Possess O, H, K antigens
infections • Biochemical reactions
Wound infections - Fermentation of glucose, lactose, trehalose,
Septicemia
and xylose
Bacteriuria
- Indole production from tryptophanase
- Glucose fermentation via mixed acid
BACTERIAL SPECIES DISEASES pathway
Serratia spp. Opportunistic and ◼ Methyl red (MR) Positive
hospital-acquired - Dose not produce H2S, DNase, urease, or
infections PAD
Wound infections - Citrate negative
Septicemia ◼ Indole; MR; Voges-Proskauer (VP);
Bacteriuria Citrate: IMVC
- Positive; positive; negative; negative
Proteus spp. Bacteriuria
Wound infection COLONIES GROWTH OF E.COLI
Septicemia

BACTERIAL SPECIES DISEASES

Povidencia spp. Opportunistic and
hospital-acquired
infections
Wound infections
Septicemia
Bacteriuria

Morganella spp. Opportunistic and

hospital-acquired
infections

BACTERIAL SPECIES DISEASES

Yersinia
Y. pestis Plague

Y. pseudotuberculosis Mesenteric adenitis

Diarrhea

Y. enterocolitica Mesenteric adenitis

Diarrhea

Erwinia spp. Wounds contaminated

with soil or vegetation

Pectobacterium spp. Wounds contaminated

with soil or vegetation
 FEATURES OF PATHOGENIC E. COLI CLINICAL TYPES OF E.COLI
TYPE VIRULENCE
FACTORS
RELEVANT
DISEASE
RELEVANT
SEROTYPES
LABORATORY
TESTS • Uropathogenic E. coli
Uropathogeni - Most common cause of UTIs in humans
c E. coli
UPEC P pilus/pap UTIs ◼ Pili allow attachment to epithelium
pili, type 1 ◼ Cytolysins can kill immune cells and
fimbriae
inhibit phagocytosis
Afa/Dr ◼ Aerobactin allows bacteria to chelate
DAEC* adhesions UTIs
iron
Enteric • ETEC
pathogens
- Diarrhea in infants and adults in tropics and
EPEC Pathogenecit Infantile O55:NM HeLa cell subtropics
y islands diarrhea O55:H6 adherence
O111:N assay, DNA - Traveler’s diarrhea (requires large inoculum
M probes 106-1010) lasts 1 to 5 days
O111:H2
O114:N
- Contaminated food and drink
M - Colonize the small intestine but adhesive
EHEC Shiga Hemorrhag O114:H2 SMAC
toxin/verotox ic diarrhea, plates, MUG
fimbriae
in colitis, HUS O157:H7 - Release toxins
O157:N
M
◼ Heat labile toxin (LT): A and B subunits; B
O26:H11 binds GM1 ganglioside receptor, and A
O104:H2
1
acts on adenyl cyclase, converting
O111:H2 DNA probes adenosine triphosphate (ATP) to cyclic
EIEC Invasin Dysentery O111:H8
O113:H2
adenosine monophosphate (cAMP),
1 which leads to hypersecretion of fluids.
O118:H2 Immunoassa ◼ Heat-stable toxin (ST) stimulates
ETEC LT, ST Traveler’s ys for LT or
diarrhea O124:H3 ST guanylate cyclase, increasing cyclic
0 guanosine monophosphate (cGMP),
O143:N
M leading to hypersecretion
O164:N • EPEC
M
- Infantile diarrhea children less than 1year
O6:NM - Nurseries and daycares
O6:H16
O8:H9 - Due to adhesions
O25:NM • Symptoms
O27:NM
O63:H12 - Low-grade fever
- Malaise
Enteroadhere
nt E. coli - Vomiting
AAF fimbriae Persistent O44:H18
- Diarrhea
EAEC
Afa/Dr pediatric ◼ Large amount of mucus without
adhesions, diarrhea
AIDA-1,
apparent blood
pathogenicit • EIEC
y islands
- Affects adults and children
DAEC Pediatric HeLa cell ◼ Rare in the United States
diarrhea, adherence - Dysentery with direct penetration, invasion,
UTIs assay, DNA
probes and destruction of intestinal mucosa
Extraintestina - Very similar to shigellae (coming up soon) but
l Pathogens
require higher amounts of inoculum
Capsule Septicemia K1 - Watery diarrhea with scant stool, pus, mucus
and
meningitis and blood
- Nonmotile and do not ferment lactose (thus
very similar to shigellae)
• EHEC - Watery diarrhea with symptoms lasting
- Hemorrhagic diarrhea, colitis more than 2 weeks
- Hemolytic-uremic syndrome (HUS) - Commensals from colon with special
◼ Low platelets, hemolytic anemia, adherence adaptations for UTIs
kidney failure ◼ Described from culture with Hep2
- Watery diarrhea progressing to bloody cells causing aggregative “stacked-
diarrhea without pus bricked” pattern
◼ Can be fatal
EXTRAINTESTINAL INFECTIONS OF E.COLI
➢ Young children and elderly
◼ Undercooked meats, unpasteurized • Septicemia and meningitis
milk, apple cider - Most common in neonates and very
• E.coli O157:H7 young children
- Toxins produced by shiga toxigenic E.coli - Gain infection just before or during
(STEC) delivery, and when infections involve the
◼ Verotoxin I (phage mediated) amniotic fluid
cytotoxin - Capsule antigen K1
➢ Damages Vero cells (African green ◼ Predisposition for meningitis
monkey kidney cells)
OTHER ESCHERICHIA SPECIES
➢ Also known as Shiga toxin (Stx)
➢ Toxin neutralized by Shiga toxin • Escherichia hermanii
antibodies - Yellow-pigmented
◼ Verotoxin II ◼ Isolated from cerebrospinal fluid
➢ Not neutralized by Shiga toxin (CSF), wounds, and blood
antibodies ◼ Isolated from foodstuffs such as raw
• Can screen O:157H7 E. coli on sorbitol- milk and beef
containing MAC (SMAC) plate - Clinical significance still not fully
- Does not ferment sorbitol and is MUG established
negative • E. vulneris
◼ Now emergent phenotype is sorbitol - Newest member
fermenting and nonmotile ◼ Wound infections
(O157:NM)
COMPARISON OF E. HERMANII AND E. VULNERIS
• Diagnosing
- Identifying serotype and comparing to
known Shiga toxin–producing strains
- Screening stool filtrates for toxin
- Demonstrate a fourfold increase in toxin-
neutralizing antibody titer
- Enteroadherent E. coli
◼ Two types
- DAECA
◼ UTIs
◼ Diarrhea
• EAEC
- Adheres to the surface of the small
intestine
KLEBSIELLA, ENTEROBACTER, SERRATIA, PANTOEA, DIFFERENTIATION OF COMMON SPECIES WITHIN
CRONOBACTER, AND HAFNIA THE GENUS KLEBSIELLA

• Tribe Klebsielleae K. PNEUMONIAE SUBSP. PNEUMONIAE

• Opportunistic and nosocomial infections TESTS OR SIGN %+ (% +)
SUBSTRATE
• Characteristics Urease + 95.4 (0.1)
- Simmon’s citrate positive Indole - 0
- Potassium cyanide broth negative Methyl red -or+ 10
Voges-proskauer + 98
- No H2S production
Citrate (simmons) + 98 (0.6)
- No deamination of phenylalanine Gelatin ( 22 C) - 0 (0.2)
- Generally urease negative Lysine + 98 (0.1)
- Indole negative (usually); MR negative decarboxylase
Malonate + 92.5
- VP positive
Mucate + 90
- Thus IMVC: negative; negative; positive; Sodium alginate + or (+) 88.5 (9.2)
positive (utilization)
Gas from glucose + 96
KLEBSIELLA SPECIES Lactose + 98.7 (1)
Dulcitol -or+ 30
• K. pneumoniae Organic Acid
- Most common isolate Media
◼ Particularly if gram-negative rods in
Citrate + or - 64.4
lower respiratory tract infection of D-tartrate + or - 67.1
hospitalized patients
◼ Also wound, UTIs, liver abscesses, and
K. OXYTOCA
bacteremia TESTS OR SIGN %+ (% +)
➢ Increase resistance, including SUBSTRATE
carbapenemase Urease + 90
Indole + 99
- Moist gray mucoid colonies
Methyl red - 20
- Virulence factor Voges-proskauer + 96
◼ Polysaccharide capsule Citrate (simmons) + 95
➢ Prevents phagocytosis and some Gelatin ( 22 C) - 0
antimicrobials Lysine + 99
decarboxylase
Malonate + 98
Mucate + 93
Sodium alginate nd
(utilization)
Gas from glucose + 97
Lactose + 100
Dulcitol +or- 55
Organic Acid
Media

Citrate nd
D-tartrate nd
 K. OZAENAE • K. pneumoniae subspecies rhinoscleromatis
TESTS OR SIGN %+ (% +)
- Rhinoscleroma
SUBSTRATE
Urease d 0 (14.8) ➢ Intense swelling and
Indole - 0 malformation of the entire face
Methyl red + 97.7 and neck
Voges-proskauer - 0
- Africa and South America
Citrate (simmons) d 30 (32.)
Gelatin ( 22 C) - 0 RAOUTELLA SPECIES
Lysine -or+ 40 (6.3)
decarboxylase • R. ornithinolytica
Malonate - 6
- Indole and ornithine decarboxylase
Mucate -or+ 25
Sodium alginate -or(+) 0 (11) positive
(utilization) • R. planticola
Gas from glucose d 50 (9.4) - Both species found in urine, respiratory
Lactose d 30 (61.3)
Dulcitol - 0
tracts and blood
Organic Acid
ENTEROBACTER SPECIES
Media
• E. cloacae
Citrate -or+ 18
D-tartrate -or+ 39 • E. aerogenes
- Two above are most common
• E. gergoviae
COLONIES OF K. PNEUMONIAE
- Respiratory samples, rarely blood
• E. hormaechei
• Citrate positive, potassium cyanide broth
positive
• MR negative, VP positive
• Usually produce ornithine decarboxylase
- Lysine decarboxylase produce by most
(not E cloacae or E. gergoviae)
• Infection sites
- Wound
- Urine
- Blood
• K. oxytoca
- CSF
- Similar to K. pneumoniae except indole
positive
- Affects similar sites
• K. ozaenae
- Isolated from nasal secretions and
cerebral abscesses
➢ Plasmid-mediated ESBLs
 DIAGNOSTIC FEATURES OF ENTEROBACTER Ornithine - 0
CLOACAE, ENTEROBACTER AEROGENES, AND decarboxylase
Gelatin (22 C) - 0
KLEBSIELLA PNEUMONIAE SUBSP. PNEUMONIAE Adonitol , gas d 84.4 (0.3)
Inositol + 97.2 (0.9)
E. CLOACAE
Acid
TESTS OR SIGN %+ (% +)
Gas + 92.5 (1.5)
SUBSTRATE
D-tartrate, + 95
Urease +w or - 65
Jordan’s
Motility + 95
Sodium alginate + or (+) 88.9 (8.9)
Lysine - 0
(utilization)
decarboxylase
Arginine + 97 (2)
dihydrolase
PANTOEA AND CRONOBACTER SPECIES
Ornithine + 96 (1.3)
decarboxylase
• Pantoea agglomerans
Gelatin (22 C) (+) 0 (92.4)
Adonitol , gas -or+ 21.7 (1.3) - Formerly Enterobacter agglomerans
Inositol - Similar to Klebsiella pneumoniae, so need
Acid d 13 (8) to differentiate
Gas - 4.1 (1.5)
- Primarily a plant pathogen
D-tartrate, -or+ 30
Jordan’s • Cronobacter sakazakii
Sodium alginate - 0 - Meningitis and bacteremia in neonates
(utilization) from powdered infant formula
- Also brain abscesses, respiratory and
E. AEROGENES wound infections
TESTS OR SIGN %+ (% +)
SUBSTRATE PHOTOGRAPH OF P. AGGLOMERANS
Urease - 2
Motility + 97
Lysine + 98
decarboxylase
Arginine - 0
dihydrolase
Ornithine + 98 (0.8)
decarboxylase
Gelatin (22 C) (+) or - 0 (61.2)
Adonitol , gas + 94.2
Inositol + 96.7
Acid
Gas + 93.4
D-tartrate, + 95
Jordan’s
Sodium alginate - 0
(utilization)

K. PNEUMONIAE SUBSP. PNEUMONIAE

TESTS OR SIGN %+ (% +)
SUBSTRATE
Urease + 95.4 (0.1)
Motility - 0
Lysine + 98 (6.3)
decarboxylase
Arginine - 0
dihydrolase
 PHOTOGRAPH OF C. SAKAZAKII • S. plymuthica
- Red pigment when growing at room
temp.
• S. ficaria
• S. entomophilia
• S. fonticola
COLONIES OF SERRATIA

SERRATIA SPECIES

• Serratia species HAFNIA

- Positive ONGP (slow lactose) • H. alvei
- Exception is S. fonticola - Linked to gastroenteritis
• DNase - Delayed citrate reaction
• Highly resistant to antimicrobials • H. alvei biotype 1
• S. marcescens - Grows in beer wort
- Prodigiosin red pigment when growing at ➢ Has not been isolated clinically
room temp
- Usually most clinically relevant PROTEUS SPECIES
- Nosocomial infection of the urinary tract, • P. mirabilis and P. vulgaris
respiratory tract, bactermia - P. mirabilis most commonly isolated
• S. liquefaciens - Isolated from urine, wounds, ear, and
• S. rubidaea blood infections
- Red pigment when growing at room - Separate the tribe by positive
temp. phenylalanine deaminase
• S. odrifera - Generally produce swarming on
- Dirty, musty odor laboratory media
- Two biogroups ➢ Burnt chocolate odor
◼ Biogroup 1: respiratory - Lactose negative and produce urease
➢ Positive to sucrose, raffinose, - Produce H2S
ornithine, and indole (60%) ➢ Occasional exceptions for P.
◼ Biogroup 2: blood and CSF vulgaris
➢ Negative for sucrose, raffinose, • P. mirabilis
ornithine, positive for indole - Indole negative, ornithine decarboxylase
(50%) positive
• P. vulgaris
- Indole positive, ornithine decarboxylase
negative
- Sucrose fermentation = A/A with H2S on
triple sugar on (TSI)
 SWARMING PROTEUS DIFFERENTIATING CHARACTERISTICS OF PROTEUS,
PROVIDENCIA, AND MORGANELLA
TEST PROTEUS PENNERI
Indole -
Methyl red +
Voges-proskauer -
Simmons citrate -
Christensen urea +
H2S (TSI) -(70%)
Ornithine decarboxylase -
MORGANELLA SPECIES Phenylalanine deaminase +
• M. morganii Acid produced from:
Sucrose +
- UTI infections
Mannitol -
- Possible agent of diarrhea Salicin -
- Similar to Proteus Adonitol -
• Five species Rhamnose -
- Two major pathogens Maltose +
◼ P. alcalifaciens may also be a Xylose +
pathogen Arabitol -
Swarms +
• P. rettgeri
- Urinary tract pathogen
- Occasionally nosocomial outbreaks TEST PROTEUS MIRABILIS
• P. stuartii Indole
Methyl red +
- Nosocomial burn until outbreaks
Voges-proskauer -or+
- Also isolated in urine cultures Simmons citrate +or(+)
- No swarming Christensen urea +or(+)
• Both have high resistance to antimicrobials H2S (TSI) +
Ornithine decarboxylase +
Phenylalanine deaminase +
Acid produced from:
Sucrose d
Mannitol -
Salicin -
Adonitol -
Rhamnose -
Maltose -
Xylose +
Arabitol -
Swarms +
 TEST PROTEUS VULGARIS TEST PROVIDENCIA STUARTII
Indole + Indole +
Methyl red + Methyl red +
Voges-proskauer - Voges-proskauer -
Simmons citrate d Simmons citrate +
Christensen urea + Christensen urea -or+
H2S (TSI) + H2S (TSI) -
Ornithine decarboxylase - Ornithine decarboxylase -
Phenylalanine deaminase + Phenylalanine deaminase +
Acid produced from: Acid produced from:
Sucrose + Sucrose d
Mannitol - Mannitol d
Salicin d Salicin -
Adonitol - Adonitol -
Rhamnose - Rhamnose -
Maltose + Maltose -
Xylose +or(+) Xylose -
Arabitol - Arabitol -
Swarms + Swarms -

TEST PROVIDENCIA TEST PROVIDENCIA RETTGERI

ALCALIFASCIENS Indole +
Indole + Methyl red +
Methyl red + Voges-proskauer -
Voges-proskauer - Simmons citrate +
Simmons citrate + Christensen urea +
Christensen urea - H2S (TSI) -
H2S (TSI) - Ornithine decarboxylase -
Ornithine decarboxylase - Phenylalanine deaminase +
Phenylalanine deaminase + Acid produced from:
Acid produced from: Sucrose d
Sucrose d Mannitol +
Mannitol - Salicin d
Salicin - Adonitol +
Adonitol + Rhamnose +or-
Rhamnose - Maltose -
Maltose - Xylose -or+
Xylose - Arabitol +
Arabitol - Swarms -
Swarms -
 TEST MORGANELLA MORGANII PRIMARY INTESTINAL PATHOGENS OF THE FAMILY
Indole + ENTEROBACTERIACEAE
Methyl red +
Voges-proskauer - SALMONELLA SPECIES
Simmons citrate -
Christensen urea + • Gram-negative rods, facultatively anaerobic
H2S (TSI) - • Clear, colorless, non-lactose-fermenting
Ornithine decarboxylase + colonies with black centers
Phenylalanine deaminase + - Negative for indole
Acid produced from:
- Negative for VP
Sucrose -
- Negative for phenylalanine deaminase
Mannitol -
Salicin - - Negative for urease
Adonitol - - Most produce H2S
Rhamnose - ➢ Exception Salmonella paratyphii A
Maltose - - No growth in potassium cyanide
Xylose -
Arabitol - S. ENTERICA
Swarms -
• Six subspecies in S. enterica
• S. typhi, S. choleraesuis, S. paratyphi
EDWARSIELLA - Differentiating Salmonella subgroups and
• E. tarda is only pathogen members
- Urea negative ➢ Lysine decarboxylase rules put S.
- Lysine decarboxylase positive paratyphi
- H2S positive ➢ Ornithine decarboxylase rules out
- Indole positive S. typhi
- No growth on citrate ➢ Trehalose fermentation rules out
S. choleraesuis
• Bacteremia and wound infections
ERWINIA AND PECTOBACTERIUM

• Plant pathogens
CITROBACTER SPECIES

• Identifying Citrobacter
- Weak urease activity
- Ferment lactose
- Grow on simmon’s citrate
- MR positive
• Similar to Salmonella
- C. freundii: most hydrolyze urea but do
not decarboxylate lysine
- Salmonella: most do not hydrolyze urea
and most decarboxylate lysine
 BIOCHEMICAL DIFFERENTIATION OF SELECT S. ENTERICA
SALMONELLA
• Virulence factors
TEST S. SEROTYPE - Fimbriae required for adhesion to
CHOLERAESUIS intestine
Arabinose fermentation - - Ability to traverse intestinal mucosa
Citrate utilization V
- Enterotoxin involved in gastroenteritis
Glucose gas production +
H2S (TSI) V ANTIGENIC STRUCTURES
Lysine decarboxylase +
Ornithine decarboxylase + • O and H antigens
Rhamnose fermentation + • H antigen
Trehalose fermentation - - Antigens occur in phases (Phase I and
Phase II).
TEST S. SEROTYPE PARATYPHI - Phase I antigens are not always present,
Arabinose fermentation + but if they are they can identify a
Citrate utilization - particular serotype
Glucose gas production + - Phase II are nonspecific
H2S (TSI) -
• Capsular antigen Vi similar to K capsule
Lysine decarboxylase -
Ornithine decarboxylase + antigen
Rhamnose fermentation + - Prevents phagocytosis
Trehalose fermentation +
Antigenic Structures of Salmonellae Used in
Serologic Typing
TEST S. SEROTYPE TYPHI
Arabinose fermentation -
Citrate utilization -
Glucose gas production -
H2S (TSI) +
Lysine decarboxylase +
Ornithine decarboxylase -
Rhamnose fermentation -
Trehalose fermentation +

TEST OTHER
Arabinose fermentation +
Citrate utilization +
Glucose gas production +
H2S (TSI) +
Lysine decarboxylase +
Ornithine decarboxylase +
Rhamnose fermentation +
Trehalose fermentation +
 CLINICAL INFECTIONS ◼ Hemorrhage and perforation of the
bowel
• Acute gastroenteritis or food poisoning with
• Nontyphoidal bacteremia
vomiting and diarrhea
- Prolonged fever with intermittent
- Caused by organism (colony-forming
bacteremia
units [CFUs] of 10^6 organisms), not
- Two groups
really by toxin
◼ Young children: fever and
◼ Poultry, milk, eggs, egg products, and
gastroenteritis with brief episodes of
pet handling
bacteremia
◼ Recently other foods such as peanut
◼ Adults: transient bacteremia during
butter, cantaloupe, cereals, dog food
episodes of gastroenteritis or
and tomatoes.
develop symptoms of septicemia
• Can develop into a carrier state and pass to without gastroenteritis
others
- Organisms are in gallbladder Culture and Serologic Diagnosis of Typhoid Fever
• Nontyphoidal bacteremia
• Typhoid fever
- Caused by S. typhi
• Other enteric fevers
- S. paratyphi
- S. choleraesius
TYPHOID FEVER DISEASE COURSE
• Ingestion of organism
• Development of fever, malaise, anorexia,
lethargy, myalgia, and a dull headache
- Looks like the flu CHARACTERISTICS OF SHIGELLA SPECIES
• Reach the small intestine, invade and
penetrate the intestinal mucosa • Nonmotile
- Leads to constipation • Generally do not produce gas from glucose
• Enter the lymphatic system and are - Except some types of S. flexneri
sustained in mesenteric lymph nodes • No urease production
• Seed the bloodstream • No H2S in TSI
- Spread to spleen, liver, and bone • No decarboxylation of lysine
marrow • Cannot utilize acetate or mucate as carbon
- Engulfed by monocytes and grow source
intracellularly • S. sonnei is positive for ONPG and ornithine
• Released into the blood stream decarboxylase.
- Prolonged bacteremia • S. flexneri is negative for these tests
- “Rose spots” in the periumbilical region
• Invasion of the gallbladder and Payer’s
patches; release of bacteria into the bowel
via the biliary duct
- Gallbladder is the foci in long-term
infections.
- Severe infections can cause necrosis of
gallbladder and/or Peyer’s patches.
 COLONIES OF S. SONNEI
BIOCHEMICAL AND SEROLOGIC DIFFERENTIATION
OF SHIGELLA SPECIES
TEST S. DYSENTERIAE
Mannitol fermentation -
ONPG V
Ornithine decarboxylase -
Serogroup A

TEST S. FLEXNERI
Mannitol fermentation +
ONPG -
Ornithine decarboxylase -
ANTIGENIC STRUCTURES Serogroup B

• O antigens separated by serologic grouping

• K antigens TEST S. BOYDII
Mannitol fermentation +
- Must be removed to type O antigen
ONPG V
- Heat labile
Ornithine decarboxylase -
• Lack H antigens Serogroup C
- Nonmotile

TEST S. SONNEI
SHIGELLA SPECIES Mannitol fermentation +
ONPG +
• S. dysenteriae (group A) Ornithine decarboxylase +
- Most serious infection (developing Serogroup D
countries)
• S. flexneri (group B)
- Second most common isolate in the
United States
- Associated in men who have sex with
men and in young adults ~25 years old
• S. boydii (group C)
- More common in developing countries
• S. sonnei (group D)
- Most common isolate in the United
States
 CLINICAL INFECTION OF SHIGELLA • Primarily affects children between 1 to 5
years and adults with underlying disease
• Dysentery
• Y. pseudotuberculosis
- Local inflammation, fever, chills,
- Rodents, farm animals, birds
shedding of intestinal lining, mucus,
- Causes caseous swellings called
blood, ulcer formation, tenesmus
psuedotubercles in mesenteric lymph
(possible rectal prolapse)
nodes
- S. dysenteriae (most severe)
• Ingestion of fecal material
• Usually nonfatal and self-limiting in
developed countries DIFFERENTIATING YERSINIA SPECIES
• Low inoculum: 10 to 200 organisms cause
• Cefsulodin, irgasan, novobiocin (CIN) agar
disease
and Yersinia-selective agar (YSA) help isolate
- Spread by fecal-oral route or by flies
Yersinia species
- Higher risk
- Now CIN II, which can also isolate
◼ Day care centers, crowded
Aeromonas
populations, anal-oral sex
◼ Children under 10 and infants under • Y. pestis
1 are particularly susceptible. - Negative for motility above 22°C
- Negative for ornithine decarboxylase
• Infection results from penetration of the
mucosal epithelium. • Y. enterocolitica
- Production of Shiga toxin produces some - Positive for motility at 25°C (room
of the symptoms temperature)
- Positive for ornithine decarboxylase
YERSINIA SPECIES • Y. pseudotuberculosis
- Positive for motility at 25°C (room
• Y. pestis
temperature)
- Plague
- Negative for ornithine decarboxylase
◼ Bubonic, septicemic, and pneumonic
forms
• Transmitted through bite of infected fleas
- Pneumonic plague through respiratory
droplets
• Gram-negative coccobacilli resembling a
safety pin appearance (bipolar staining)
- Preferential growth at 25°to 30°C
• Y. enterocolita
- Acquired from contact with swine, cats,
dogs
◼ Ingestion of contaminated food that
contacted fecal material
◼ Refrigeration is ineffective because it
survives in cold temperatures.
• Symptoms
- Acute gastroenteritis, simulating
appendicitis
- Arthritis
- Erythema nodosum
◼ Tender red nodules with itching and
burning on lower legs (shins)
DIFFERENTIATION OF SELECTED SPECIES WITHIN TEST Y. PSEUDOTUBERCULOSIS
THE GENUS YERSINIA Indole -
Methyl red +
TEST Y. PESTIS Voges-proskauer
Indole - 25 C -
Methyl red + 37 C -
Voges-proskauer Motility
25 C - 25 C +
37 C - 37 C -
Motility B-Galactosidase +
25 C - Christensen urea +
37 C - Phenylalanine deaminase -
B-Galactosidase - Ornithine decarboxylase -
Christensen urea + Acid produced from:
Phenylalanine deaminase - Sucrose -
Ornithine decarboxylase - lactose -
Acid produced from: Rhamnose +
Sucrose - Melibiose +
lactose - Trehalose +
Rhamnose - Cellobiose -
Melibiose -
Trehalose -
Cellobiose - LABORATORY DIAGNOSIS OF ENTEROBACTERIACEAE
• Specimen collection and transport
TEST Y. ENTEROCOLITICA • Isolation and identification
Indole d - Direct microscopic examination
Methyl red + ◼ Not very helpful in differentiating
Voges-proskauer species
25 C d - Culture
37 C - ◼ Generally grow quickly and easily
Motility
- Identification
25 C +
• Screening stool cultures for pathogens
37 C -
B-Galactosidase +
- Separate pathogens from normal flora
Christensen urea +
Phenylalanine deaminase -
Ornithine decarboxylase + LYSINE IRON AGAR (LIA) AND TSI SCREENING
Acid produced from:
Sucrose +
lactose -
Rhamnose -or+*
Melibiose -or+*
Trehalose +or-
Cellobiose +
 HEKTOEN ENTERIC AND XYLOSE LYSINE SCHEMATIC DIAGRAM FOR ENTERICS
DEOXYCHOLATE (XLD) AGAR

CHROMagar™

SEROLOGIC GROUPING
IDENTIFICATION
• Serologic grouping
• Determine if Enterobacteriaceae
- Salmonella
- Gram-negative
◼ 60 types of O antigens
- Oxidase negative
➢ 95% are serogroups A through E1
◼ Except for Plesiomonas shigelloides
◼ Direct or latex agglutination tests for
◼ Always use young colonies from
serogroup
sheep blood agar (SBA) plates
◼ Shigella
- Ferment glucose
➢ A through D serogroups
- Reduce nitrate to nitrite
◼ Except Photorhabdus and
Xenorhabdus