ENTEROBACTERIACEAE

ENTEROBACTERIACEAE
Gr (-), non sporeforming, FA
Most are present in the intestinal tract as commensal flora (EXCEPT
SALMONELLA, SHIGELLA AND YERSINIA)
CM: Gr (-) straight rods or coccobacilli with rounded ends
CULTURE: large, smooth, Gray colonies (except Klebsiella and Enterobacter) ;
nonhemolytic (except E. coli (B-hemolytic)).
Biochemical tests: catalase (+); cytochrome oxidase (-) (except P. shigelloides)
MOTILE with peritrichous flagella except Klebsiella, Shigella and Yersinia.
NONENCAPSULATED except Klebsiella, and Enterobacter.
All members ferment glucose and reduce nitrate to nitrite.
Some organisms may grow at low temperatures = 1-5 C (Serratia and Yersinia).
 ENTEROBACTERIACEAE
Two groups:
1.) Opportunistic - Proteus, Enterobacter, Citrobacter, Klebsiella, Serratia
2.) Overt/True - Salmonella, Shigella, Yersinia

Extended-spectrum-B-lactamase prod. Enterobacteria -> E. coli, K. pneumonia, K. oxytoca

Isolated fr. Urinary tract and cause Bacteremia -> E.coli, P. mirabilis, K. pneumoniae

Antibiotic resistant genera -> Serratia, Enterobacter, Citrobacter

Antigenic determinants for serologic ID.

Somatic “O” antigen
Flagellar “H” antigen
Capsular “K” antigen (Found as K1 antigen (E.coli) and Vi antigen (S. typhi))
 ESCHERICHIA COLI
Colon microbiota
primary marker of fecal contamination in water quality testing
Virulence factors:
adhesive fimbriae and sex pili
O, H & K antigens
E.coli Strains:
A. Uropathogenic E. coli
B. Gastrointestinal E. coli
C. Extraintestinal E. coli
ESCHERICHIA COLI
Culture:
Flat, dry, pink on (Mac)
B-hemolytic on (BAP)
Greenish metallic sheen (EMB)
IMVC reaction: + + - -
Indole (+)
MR (+) VP (-)
Citrate (-)
H2S, Dnase, urease, PAD (-)
TSI reaction: A/A, (+) gas, (-) H2S
LIA: A/A or K/A
E.COLI STRAINS
Infantile Diarrhea
Enteropathogenic (EPEC)
(attach to brush border of intestinal cells [adhesive])

Traveler’s Diarrhea / Montezuma’s Revenge

At risk: Achlorydia
Enterotoxigenic (ETEC) VF: heat stable (ST) and heat labile (LT) enterotoxin
10^6 to 10^10 organisms (infective dose)
One of the major causes of infant bacterial diarrhea

Shigella-like infection; Watery diarrhea with WBCs

Enteroinvasive (EIEC)
10^6 infective dose
E.COLI STRAINS
Hemorrhagic colitis
Hemolytic Uremic Syndrome (HUS)
Bloody diarrhea (no WBCs)
Enterohemorrhagic Verotoxin I and II (Cytotoxin) <- I: Shiga-like toxin (STX
(EHEC serotype 0157:H7) T1)

SMAC (sorbitol) – stool culture

(-) ferment sorbitol in 48 hrs

-Watery diarrhea and UTI

Enteroadherent -UTI
Fimbriae (stacked brick)
a. EAEC
b. DAEC (diffusely) EAEC: Adheres to Hep2 Cells froming clumps of bacteria
“stacked brick” appearance
 E.COLI STRAINS
Uropathogenic
Most common cause of UTI
+ Pyelonephritis (ICP)
UPEC: P pilus/ pap pili, type 1 fimbriae
VF: Pili; T1 fimbriae, Cytolysins (Kill immune effector cells) ,
DAEC: Afa/Dr adhesions Aerobactins (chelates iron)

Extraintestinal E. coli - Septicemia and neonatal

meningeal infection ; VF: Capsular antigen K1

yellow-pigmented organism in CSF, wound & blood

Escherichia hermannii
(more prevalent)

Escherichia vulneris yellow-pigmented organism

 KLEBSIELLA
Microbiota of GIT
Gr (-) bacilli, KCN (+) VP (+) MAC (LF)
Fermentation of glucose, lactose, trehalose & xylose
TSI reaction: A/A, (+) gas, (-) H2S
LIA: A/A
IMVC reaction: - - + +
KLEBSIELLA
KLEBSIELLA
IMVC Reactions
 ENTEROBACTER
isolated from wounds, urine, blood, and CSF
Culture (MAC) - Pink mucoid (LF)
Significant species: E. aerogenes, E. cloacae (common isolate), E.
gergoviae and E. hormaechei
Fermentation of glucose, lactose, trehalose & xylose
TSI: A/A, (+) gas, (-) H2S (E. aerogenes and E. cloacaae)
LIA: K/K
IMVC: - - + +
Ornithine decarboxylase & lysine decarboxylase (+) EXCEPT E.
gergoviae or E. cloacae
Urease & motility (+)
ENTEROBACTER
 ENTEROBACTER
DIFFERENTIAL DIAGNOSIS
 SERRATIA
opportunistic pathogens associated with outbreaks in health care
settings (UTI & RTI)
bacteremic outbreaks in nurseries and cardiac surgery and burn units
Slow/late lactose fermenter EXCEPT S. fonticola
DNase, lipase, gelatinase (+)
(ONPG) test (+)
S. marcescens, S. rubidaea & S. plymuthica -> Prodigiosin at 25’C; pink
to red pigment in RT
SERRATIA
 SALMONELLA
Most serious pathogenic enterobacteria for humans
ingestion of contaminated animal food products or improperly cooked
poultry, milk, eggs and dairy products. It may also be transmitted by
"human carriers."
Causing Enteric Fever (Typhoid fever) and Acute Gastroenteritis (food
poisoning)
Carrier state following Salmonella infection
VF: Fimbriae and Enterotoxin
Ag: O, H, Vi antigen
all are motile except S.Pullorum and S. Gallinarum
all produce gas except S. Gallinarum and S. Typhi.
All produce H2S except S. Paratyphi A.
 SALMONELLA
Gram (-) facultatively anaerobic bacilli
MAC: clear, colorless, non–lactose fermenting colonies
XLD & HE: colonies with black centers (H2S +)
Bismuth Sulfide Agar: black colonies with metallic sheen
Non-lactose fermenter
Indole, VP, PAD, Urease (-)
H2S (+) EXCEPT S. paratyphi
(-) growth potassium cyanide medium

SALMONELLA
Culture:
clear, colorless colonies
- Mac Conkey
colored colonies with black centers
= media with H2S (HE, BSA and XLD)
colorless colonies with black centers
- SSA
IMVC -+-+ [-+-- (S.Typhi)]
TSI reaction: K/A, (+) gas, (+) H2S
K/A, (-) gas, (+) H2S -> (S. Typhi)
Somatic O antigen
Heat stable
Lipopolysaccharide (cell wall)
Flagellar H
Heat labile
Phase I flagellar antigen:
specific phase; occur in few strains
Phase II flagellar antigen:
non-specific phase; occur in may strains
(Agglutinate only with
heterologous antisera)
Capsular K antigen (Vi)
For ID of Salmonella serotype Typhi &
Salmonella serotype Choleraesuis
SALMONELLA INFECTIONS
 SALMONELLA
Carrier state :
Site of chronic carriage: Gallbladder
Organisms secreted in the feces continuously or intermittently (important
source of infection)
Treatment:
- Antimicrobial therapy
- Cholecystectomy
 SALMONELLA
BIOCHEMICAL DIFFERENCES OF SELECTED SALMONELLA SEROTYPES
 SHIGELLA
closely related to the genus Escherichia.
nonmotile
not GI microbiota
All species except S. dysenteriae are mannitol fermentation test (+)
does not utilize acetate or mucate as a source of carbon
(-) gas from glucose (except S. flexneri)
Urease: -
LDC: -
IMVC reaction: v + - -
TSI reaction: K/A, (-) gas, (-) H2S
Susceptible to disinfectants & high concentrations of acids and bile
 SHIGELLA
intracellular organism
they multiply within the cells of the colon epithelium
transmitted by four F's (fecal-oral route) -> files, fingers, food and feces
Reservoir: humans only
Culture: clear, fragile, NLF colonies
SSA: colorless colonies -> without black centers
Virulence factor: SHIGA TOXIN
SHIGELLA
SHIGELLA
 YERSINIA
Y. PESTIS “PLAGUE BACILLUS”
Class A bioterrorism agent
Xenopsylla cheopsis (flea) - only Enterobacteriaceae that is
transmitted to humans by the bite of an infected flea (vector)
Bubonic plague
"black death or 6th century pandemic."
ingestion of contaminated animal tissues and
Inhalation of contaminated airborne droplets.
multiply in the blood and lymph.
ability to survive and multiply inside phagocytes - due to yersinal
plasmid-encoded outer membrane proteins (YOPS).
 YERSINIA
"BIPOLAR STAINING or closed SAFETY PIN appearance"
short, plump rod with "bipolar staining or closed safety pin appearance"
Wayson or methylene blue stain
“Stalactite pattern” - broth
pinpoint colonies; grows best at 25° 30°C.
"stalactite pattern"- broth cultures
Virulence factors: Endotoxin, Coagulase and Fibrinolysin
TSI reaction: K/A (-) gas (-) H2S
 Y. PESTIS (PLAGUE)
High fever with painful regional lymph nodes
Bubonic/ glandular form known as buboes; swelling of axilla and groin

Bacteria spread
Septicemic form to the bloodstream

Secondary to bubonic plague or

septicemic form;
Pneumonic form
Primary infection through
Inhalation;
Fatality rate: 100% if untreated
YERSINIA
HAFNIA
Hafnia alvei (formerly Enterobacter hafniae)
H. alvei and H. alvei biotype 1
Biotype 1 grows in the beer wort of breweries and
has not been isolated clinically.
Culture:
MAC: NLF; colonies are clear and colorless
HE: Colorless
XLD: Yellow or colorless
PROTEUS
Belong to tribe Proteeae (Morganella and Providencia)
Widely disseminated in the environment, normal intestinal
microbiota, and are recognized as opportunistic pathogens
Causes: Lower and Upper Urinary Tract Infection, acute
glomerulonephritis
Isolated from urine, wound, and ear infection
PROTEUS
Species: Proteus mirabilis, Proteus vulgaris, Proteus penneri,
Proteus myxofaciens
Human pathogens: Proteus mirabilis & Proteus vulgaris
Most common isolate: Proteus mirabilis
Culture:
MAC: NLF; colonies are clear, may swarm, depending on the amount of
agar in the medium; “chocolate cake” or “burnt chocolate” odor
HE: Colorless
XLD: Yellow or colorless
PROTEUS
PROTEUS
PROVIDENCIA
Causes: Opportunistic and nosocomial infections
(involving burn units), wound infections, bacteriuria, and
septicemia.
Species: P. alcalifaciens, P. rettgeri, P. rustigianii, P.
stuartii, and P. heimbachae
PROVIDENCIA
Culture:
MAC: NLF; colonies are clear and colorless
HE: Colorless
XLD: Yellow or colorless
Biochemical Tests:
PAD test: positive
IMViC reaction: ++-+
LIA reaction: R/A
TSIA reaction: K/A, (-) gas, (-) H2S
PROVIDENCIA
Providencia rettgeri:
It is a pathogen of the urinary tract
It also causes diarrheal disease among travelers.
It is mostly resistant to antimicrobial agents.

Providencia stuartii
It is found in nosocomial outbreaks in burn units.
It is resistant to antimicrobial agents same with P. rettgeri.
Isolated in urines.

Providencia alcalifaciens
It is the most commonly isolates in fecal specimen of children with diarrhea.
MORGANELLA
Cause of neonatal sepsis and UTI
Species: M. morganii ss. morganii and M. morganii ss.
sibonii
Culture:
MAC: NLF; colonies are clear and colorless
HE: Colorless
XLD: Red or colorless
MORGANELLA
Biochemical Tests:
PDA test: Positive
LIA reaction: R/A
IMViC reaction: ++--
TSIA reaction: K/A, (+) gas, (-) H2S
Other biochemical tests: (+) Urease, KCN, and ornithine
decarboxylase
EDWARDSIELLA
Isolated from cold-blooded and warm-blooded animals.
Mode of transmission: Ingestion of food and water
contaminated with infected animal products and excreta
Species: E. tarda, E. hoshinae, and E. ictaluri
Human pathogen: E. tarda
Causes: bacteremia & wound infections
EDWARDSIELLA
Culture:
MAC: NLF
HE: Colorless
XLD: Red, Yellow, or colorless colonies, with or
without black centers (H2S)
Biochemical Tests:
Urease test: -
LDC test: +
IMViC reaction: ++--
TSIA reaction: K/A, (+) gas, (+)H2S
CITROBACTER
GIT inhabitants
Resemble E. coli on MacConkey agar and Salmonella in
biochemical tests.
Cause: Opportunistic and nosocomial infections (UTI & sepsis)
Can cause false-positive agglutination test results with
Salmonella
All species grow in Simmons citrate agar (SCA)
Common isolates: C. freundii, C. koseri, and C. braakii, and C.
farmeri
CITROBACTER
Culture:
MAC: Late lactose fermenter; therefore, NLF after 24
hours; LF after 48 hours; colonies are light pink after
48 hours
HE: Colorless
XLD: Red, Yellow, or colorless colonies, with or without
black centers (H2S)
CITROBACTER
Biochemical test:
Urease test: (+)
IMViC reaction:
C. freundii: -+-+
C. koseri: ++-+
TSIA reaction:
C. freundii: K/A or A/A, (+) gas, (+) H2S
C. koseri: K/A, (+) gas, (-)H2S
CITROBACTER
Citrobacter freundii
It can be isolated in diarrheal stool cultures
(extraintestinal pathogen)
It is associated with pneumonia, intraabdominal
abscess, and endocarditis in intravenous drug
users.
It produces group 1 cephalosporinase.
Citrobacter koseri (formerly C. diversus)
It causes outbreaks of neonatal meningitis and
brain abscess in nursery units.
CITROBACTER
PLEOSIOMONAS
Plesiomonas shigelloides
It is the only species in the genus Plesiomonas.
not part of the indigenous human microbiota and
is considered a true pathogen.
It is found in fresh and estuarine water, and it has
been isolated from warm- and cold-blooded
animals.
It often cross-agglutinates with Shigella, hence
the species name “shigelloides”.
PLEOSIOMONAS
Plesiomonas shigelloides
It is the only oxidase-positive member of the
Enterobacteriaceae.
It is motile by monotrichous or lophotichous
flagella.
It has been isolated from HIV-positive individuals
with inflammatory bowel disease.
Mode of acquisition: ingestion of undercooked
seafood (oysters and shrimps) and
contaminated water; they gain entry thru skin
cuts.
PLEOSIOMONAS
Plesiomonas shigelloides
Virulence factors: hemolysins, cytotoxins,
production of exoenzymes
Microscopy: straight bacilli which can occur
singly, in pairs, in short chains, or filamentous
Vibriostatic Test O/129: Sensitive
Related infection: secretory diarrhea, neonatal
meningitis, and septicemia.
PLEOSIOMONAS
Cultural Characteristics:
a.BAP: colonies are shiny, opaque, and non-hemolytic.
b.MAC: colonies are clear and colorless (NLF). Some
srains will grow on MAC.
c.Inositol-brilliant green-bile salt agar: Colonies
exhibit white or green to pink color while Aeromonas
species are colorless. This medium enhances the
recovery of plesimonads from specimens.
d.HEA: colonies exhibit growth.
e.TCBS: colonies do not exhibit growth.
f. Media with NaCl: Colonies do not exhibit growth.
PLEOSIOMONAS
Biochemical and Serological Characteristics
Carbohydrate fermentation test: Glucose, maltose,
trehalose
Oxidase test: positive
Decarboxylase test: positive trio decarboxylate test
Inositol fermentation: Positive
IMVIC reaction: ++--
TSIA reaction: K/A, (-) gas, (-) H2S
Antigenic determinant: Somatic O and flagellar H
NON LACTOSE FERMENTERS

Proteus

Providencia

Morganella

Salmonella

Shigella

Edwardsiella

Erwinia

Yersinia
LACTOSE FERMENTERS

Escherichia

Klebsiella

Enterobacter

Serrati

Citrobacter
 NON-ENTERIC
GASTROINTESTINAL
PATHOGENS
Vibrio, Aeromonas,
Plesiomonas, and
Campylobacter
species

By Yna Andrea Belen

 Vibrio
Family: Vibrionaeae

Environment: fresh water, brackish water, marine/salt water

Indication of Vibrio spp. infection:
— consumption of raw seafood
— immigration or foreign travel
— cholera-like or rice-water stool
— trauma incurred during contact with fresh, estuarine, or marine water or
associated products
Vibrio spp.
Characteristic: Biochemical test:
non-spore forming Catalase (-)

facultative anaerobe Oxidase (+)

halophilic Reduce nitrate to nitrite

pleomorphic gram (-) bacilli String test (+)

— 0.5% sodium desoxycholate
Vibriostatic compound O/129 (S)
— 2,4-diamino-6,7 diisopropylpteridine
Vibrio
Group 1
— V. cholera & V. mimicus
Group 2
— V. metscchnikovii
Group 3
— V. cincinnatiensis
Group 4
— V. hollisae
Group 5
— V. damsel & V. fluvialis
Group 6
— V. alginolyticus, V. parahaemolyticus, V. vulnificus & V. harveyi
Vibrio cholerae
V. cholerae subgroups:
V. cholerae O1
— Ogawa (A,B)
— Inaba (A, C)
— Hikojima (A, B, C)
V. cholerae O139

V. cholerae non-O1
— resemble V. cholerae but fail to agglutinate in O1 antisera

V. parahaemolyticus
— O and K antigens
 Vibrio cholerae
Vibrio cholerae O1: cholera, “Asiatic cholera”, or “epidemic cholera”

Enterotoxin: cholera toxin or choleragen

2 biogroups:

Classic

El Tor
 Vibrio cholerae
Vibrio cholerae non O1:
most lack the cholera toxin gene
milder form of gastroenteritis or cholera-like disease

V. cholerae serogroups O75 & O141

(+) cholera toxin gene
sporadic, cholera-like disease
 Vibrio parahaemolyticus
2nd most common vibrio causing gastroenteritis “summer diarrhea”

Pandemic strain: V. parahaemolyticus serotype O3:K6

Kanagawa phenomenon
— heat-stable hemolysin effective in high-salt mannitol medium

Wagatsuma agar
Vibrio vulnificus
2nd most serious cause of Vibrio-associated type infections
— septicemia
— wound infections

Vibrio alginolyticus
least pathogenic for human
strict halophile
eye, ear, wound or burn infections
Specimen collection:
— body fluids, pus, tissues

Transport medium: prevent dessication

Cary-Blair
glycerol: toxic for vibrios Buffered glycerol saline

Time of Collection:
ASAP in the course of illness
before antimicrobial administration

Direct microscopic examination:

— pleomorphic gram (-) bacilli
 Culture and Macroscopic examination
NA & BAP
MAC: NLF except V. vulnificus
TCBS: thiosulfate citrate bile salt sucrose
H2S indicator: ferric citrate

Sucrose fermenters:
V. cholerae, V. alginolyticus, V. fluvialis, V. furnissii, V. cicinnatiensis, V. metschnikovii and some V. vulnificus

Nonsucrose fermenters:
V. mimicus, V. parahaemolyticus, P. damsela, and most V. vulnificus strains

Enrichment medium:
— Alkaline Peptone Water with 1% NaCl pH 8.5
Biochemical tests:
150 ug vibriostatic agent O/129 (S)
String test (+)
Ferment inositol (-)
Oxidase (+)
O/F reactions (sucrose fermenter)
Vibrio
Group 1
— V. cholera & V. mimicus
Group 2
— V. metscchnikovii
Group 3
— V. cincinnatiensis
Group 4
— V. hollisae
Group 5
— V. damsel & V. fluvialis
Group 6
— V. alginolyticus, V. parahaemolyticus, V. vulnificus & V. harveyi
Vibrio
Group 1
— V. cholera & V. mimicus
Group 2
— V. metscchnikovii
Group 3
— V. cincinnatiensis
Group 4
— V. hollisae
Group 5
— V. damsel & V. fluvialis
Group 6
— V. alginolyticus, V. parahaemolyticus, V. vulnificus & V. harveyi
 Aeromonas
Characteristics:
Family: Aeromonadaceae
oxidase (+)
glucose fermenting
gram-negative bacilli
Environment: freshwater, estuarine, and marine environment

Groups:
— mesophilic group (37°C) — pyschropilic group (22°C)
A. hydrophilia A. salmonicida
A. veronii complex
A. caviae complex
 A. caviae
Gastrointestinal infections
— Acute, secretory diarrhea often accompanied by vomiting
— Acute, dysenteric form of diarrhea (similar to shigellosis)
— chronic diarrhea lasting more than 10 days
— cholera-like disease, including rice water stools
— nebulous syndrome commonly referred to as traveler’s diarrhea (similar
to enterotoxigenic E. coli)

— septicemia, meningitis, wound infections & keratitis associated with

contact lens wear
 Aeromonas
Specimen collection
No special considerations
Culture
large, round, raised, opaque colonies with an entire edge and a
smooth , often mucoid, surface
BAP (beta hemolysis): A. hydrophilia, A. veronii biovar sobria, and
A. jandaei
CIN w/ 4 ug of cefsulodin instead of 15 ug
— pink-centered colonies (mannitol fermentation)
— with uneven, clear apron resembling Y. enterocolitica
Oxidase (+)
Biochemical tests:
Indole (+)
Oxidase (+)
String test (-)
O/129 (R)
0% NaCl (+)
6% NaCl (-)
Inositol (-)
glucose fermentation (+)
Plesiomonas
Characteristics:
Family: Enterobacteriaceae
Motile, oxidase (+), glucose fermenting, facultatively anaerobe,
gram (-) bacilli
P. shigelloides
— (S) O/129

Infection:
— Gastroenteritis
wateryor secretory diarrhea
subacute or chronic disease that last from 14 days to 2-3 months
more invasive, dysenteric form
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Plesiomonas
Characteristics:
Motile: monotrichous or two to five lophotrichous flagella
P. shigelloides
— cross-agglutinate with S. sonnei, S. dysenteriae & S. boydii

Virulence factors:
O and H antigen

Microscopic examination:
Gram (-) bacillli in singly, in pairs, or in short chains or filamentous forms

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Plesiomonas
Culture and Macroscopic examinatio:
shiny, opaque, nonhemolytic colonies appear, with a slightly raised center
and a smooth and entire edge
MAC: may be LF or LLF
— Oxidase test (+)
Inositol Brilliant Green Bile Agar
— white to pink colonies
(+) growth in CIN
— colonies with an opaque apron
(-) growth in TCBS

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Biochemical tests:
maybe LF and LLF
Oxidase (+)
Inositol (+)
growth in CIN (+), opaque apron
growth in TCBS (-)
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Campylobacter & Campylobacter-Like species
Characteristics:
Previously classified under vibrios (oxidase +)
Asaccharolytic
Family: Campylobacteraceae
— Campylobacter, Arcobacter, Sulfurospirillum
Microaerophile
— 5% oxygen

Family: Helicobactereaceae
— Helicobacter and Wollnella
Campylobacter & Campylobacter-Like species
Campylobacter spp.

abortion in domestic animals

Campylobacter jejuni

most common cause of bacterial gastroenteritis

4th most common cause of foodborne gastrointestinal (Foodborne
Diseases Active Surveillance Network)

Helicobacter pylori

gastric, peptic, and duodenal ulcers, GI carcinoma

Campylobacter & Campylobacter-Like species
Specimen Collection:
C. fetus subsp. fetus
Blood culture
Campylobacter spp.
stool samples & rectal swab
Transport medium:
— Cary-Blair
— Buffered glycerol saline
H. pylori
gastric biopsy
Transport medium: Stuart medium
Tissue samples: Cysteine-Brucella broth with 20% glycerol (frozen at -70°C)
Campylobacter & Campylobacter-Like species
Culture and Macroscopic examination:
Selective media for Campylobacter spp.
Campylobacter & Campylobacter-Like species
Culture and Macroscopic examination:
CAP or Brucella agar with 5% horse red blood cells & Skirrow’s agar
— H. pylori
C. jejuni and other enteric campylobacters grow optimally at 42°C
— EXCEPT: C. fetus subsp. fetus (37°C)
Microaerophilic and capnophilic environment
Campylobacter spp.
— 5% O2, 10% CO2, and 85% N2
— EXCEPT C. rectus & C. curvus (strict anaerobes)
Helicobacter spp.
— 5% to 10% O2 and 5% to 12% CO2
Campylobacter & Campylobacter-Like species
Culture and Macroscopic examination:
GasPak jar
— for Campylocaters

Evacuation replacement system

– strict anaerobic condition
– anaerobic jar
Pressure: 15 inches Hg
Gas: 10% CO2, 90% N2
— 5% CO2, 10% H2, 85% N2
— 10% CO2, 10% H2, 80% N2
– Candle jar
— least ideal environmental condition
Campylobacter & Campylobacter-Like species
Microscopic examination:
Campylobacter spp. & Arcobacter spp.
— curved, non–spore-forming, gram (-) bacilli
• Campylobacter jejuni & other enteric campylobacter
— long spirals or ‘S’ or seagull-wing shapes
• Gram stain: stain poorly
• carbolfuchsin : recommended counter stain (safranin 2-3 mins extension)
• Hanging drop preparations or Phase contrast microscope:
“darting” motility“ single polar flagellum
Brucella tryptic soy broth not in dist. water
H. pylori
• curved, non–spore-forming, gram (-) bacilli
• multiple flagella at one pole
Campylobacter spp. Helicobacter pylori

Oxidase (+) identified by nonculture methods

(+) growth at 42° C in a Gastric biopsy
microaerophilic environment Rapid urease (+)
Hippurate hydrolysis test (+): C. jejuni incubated at 37° C for 2 hours
Urea breath test
oral administration of 13C- or 14C-
labeled urea
release of 13CO2 or 14CO2
detected in the exhaled breath by a
scintillation counter
Anna Czarina Janile Chris Yna Andrea Belen Ariel Bicaldo Jr.
Aliabo Belbis
THANK YOU
NON-FERMENTATIVE
GRAM-NEGATIVE
BACILLI
by Ariel
Bicaldo Jr.
 CONTENT
Non-fermentative
Gram-negative bacilli

1 2
Pseudomanas Burkholderia
3
Stenotrophomonas
4
Acinetobacter
P. aeruginosa B. mallei S. maltophilia A. baumanni
P. fluorescens B. pseudomallei A. iwoffi
P. putida B. cepacia A. haemolyticus
P. stutzeri
P. mendocina Alkaligenes faecalis Moraxella lacunata
NONFERMENTING GRAM-NEGATIVE BACILLI
GENERAL CHARACTERISTICS
Grow in MacConkey as colorless colonies
Fail to acidify O-F media, overlaid with mineral oil
Fail to acidify TSI Agar
Most isolates is Oxidase (+)
Most have a characteristic pigment and odor
Most are multi-drug resistant (MDR)
NONFERMENTING GRAM-NEGATIVE BACILLI

NOSOCOMIAL INFECTIONS:
P. aeruginosa
Acinetobacter
S. maltophilia
NONFERMENTING GRAM-NEGATIVE BACILLI
GENERAL CHARACTERISTICS
Some oxidize CHO to derive energy for their
metabolism: Oxidizer
Some are inert or biochemically inactive:
nonoxidizers or asaccharolytic
Most are Oxidase (+): differentiate from
Enterobacteriaceae
PSEUDOMONAS
AERUGINOSA
 PSEUDOMONAS
AERUGINOSA
•Obligate aerobes
•Motile and rod shaped
•Sweet and grape-like or corn-taco like odor
resulting from production of 2-
aminoacetophenon
•Grows well at 37C, its growth at 42C will
differentitate it from other Pseudomonas sp.
•Oxidase positive
•Does not ferment carbohydrate
•TSI K/K or K/NC (no change)
 PSEUDOMONAS
AERUGINOSA

•Acetamide Positive

•Grows in Cetrimide
(cetyltrimenthylammonium bromide) Agar

- ‘’serrated confluent growth’’

 PSEUDOMONAS
AERUGINOSA
•Identifying Characteristics:

Pigments
- Fluorescein (Pyoverdin) – yellow
-Pyocyanin – blue
-Pyorubin – red
-Pyomelanin – brown/black
 PSEUDOMONAS
AERUGINOSA
•Produces infection of wound and burns, giving
rise to blue-green pus
•Meningitis
•UTI
•Pneumonia
•Cystic fibrosis patients
•Sepsis
•Mild otitis externa/media – ‘’swimmer’s ear’’
•Skin lesion – ecthyma gangrenosum,
Jacuzzi hot tub / Whirlpool dermatitis
 PSEUDOMONAS
AERUGINOSA

•Keratitis – from contamination of contact lens

solution

•Acanthamoeba – Keratitis due to contamination of

contact lens, or direct inoculation to eyes
 PSEUDOMONAS
FLUORESCENS &
PSEUDOMONAS PUTIDA
Characteristics:
•Produce pyoverdin but not pyocyanin
•Grows at 42 C
•Cannot reduce nitrate to nitrogen gas
•Produce acid from xylose: separate them from
other flourescent pseudomonads

•Gelatin hydrolysis
• Pseudomonas fluorescens(+)
• Pseudomonas putida (-)
 PSEUDOMONAS
STUTZERI

Characteristics:

•Wrinkled, leathery, adherent colonies producing

light-yellow or brown pigment

ADH (-)
Starch hydolysis (+)
6.5 NaCl (+)
Nitrate to nitrite reduction (+)
 PSEUDOMONAS
MENDOCINA

Characteristics:

•Nonwrinkled, flat colonies that may appear with

a yellowish-brown pigment

•Oxidase and ADH (+)

•Not produce pyoverdin
•Acetamide & starch hydrolysis (-)
•Motile (single polar flagellum)
•Oxidizes glucose and xylose
BURKHOLDERIA
BURKHOLDERIA
MALLEI

Causes: Glanders
Nonmotile
BURKHOLDERIA
PSEUDOMALLEI

(WHITMORE’S BACILLUS’’)

Causes ‘’Melioidosis’’ (Vietnamese

Time Bomb)

Identifying characteristics:
Bipolar staining (safety pin) in
Gram Stain
BURKHOLDERIA
PSEUDOMALLEI

(WHITMORE’S BACILLUS’’)

Smooth to wrinked colonies in

BAP

Wrinkled and deep pink in

Ashdown media

Earthy ‘’odor’’
BURKHOLDERIA CEPACIA

•Oxidase and LDC (+)

•Motility = Lopotrichous

•Oxidative-Fermentative
Polymyxin B. Bacitracin lactose
(OFPBL) / PC Agar = Yellow

•Pink colony on Mac (lactose oxidizer)

•‘’Earthy or dirt like odor’’

BURKHOLDERIA CEPACIA

•Heart valve

•Endocarditis

•Cause onion bulb rot in plants

•Foot rots in humans

•2nd most common infection in cystic

fibrosis patients
STENOTROPHOMONAS
MALTOPHILIA
STENOTROPHOMONAS
MALTOPHILIA
•OLD NAME: XANTHROMONAS
•OXIDASE (-), DNASE (+)
•OXIDIZER OF GLUCOSE, STRONG
OXIDIZER OF MALTOSE
•LAVANDER GREEN COLONY
•‘’AMMONIA-LIKE ODOR’’
•TSA = YELLOW PIGMENT
•CAUSES WOUND INFECTION
(FARMING EQUIPMENT)
- 3RD MOST COMMON CAUSE OF
NOSOCOMIAL INFECTION
STENOTROPHOMONAS
MALTOPHILIA
IDENTIFYING CHARACTERISTICS:

LAVANDER GREEN
PIGMENT IN BAP

AMMONIA-LIKE SMELL

OXIDIZES GLUCOSE: W(+)

ACINETOBACTER
SPP.
 ACINETOBACTER SPP.

•TSI: K/K, Oxidase (-), catalase (+)

•Nonmotile
•MaC = Purple colony
•Blue- grey (cornflower blue) in EMB
•Drug resistant
•Associated infections and diseases: UTI,
wound, diarrhea

•A.baumanni– Oxidizer/Saccharolytic, grows

at 42 C
•A.iwoffi– Non-oxidizer/ Assachrolytic
•A. haemolyticus– Beta-hemolytic strain
ALKALIGENES
FAECALIS &

MORAXELLA
LACUNATA
ALKALIGENES
FAECALIS
•Oxidase, Catalase (+)
•Motility: Peritrichous
•Apple-like ‘’fruity’’ odor
•O-F (-/-), Assacharolytic
•Associated infections and
diseases: UTI, wound, diarrhea
MORAXELLA
LACUNATACharacteristic:
Causes: Blepharoconjunctivitis
Oxidase (+), Catalase (+)
O/F = -/- (assachorolytic)
(-) growth in MAC

Gram (-) coccobacilli

Mistaken as Neisseria
WHICH PSEUDOMONAS IS USUALLY ASSOCIATED WITH A LUNG
INFECTION RELATED TO CYSTIC FIBROSIS?

A P. PUTIDA

B P. AERUGINOSA

C P. FLUORESCENS

D BURKHOLDERIA PSEUDOMALLEI