Enterobacteriaceae

• Prepared by
• Yasser Riyadh Hazim • Alaa Elias Hameed
• Yasir Saadi Mahmood • Ali Amer Mohammad
• Mohammed Nawfal Abdulmajeed
• Ahmad Ibrahim Saleh
GENERA AND SPECIES TO BE CONSIDERED
GENERAL CHARACTERISTICS

• Gram-negative bacilli or coccobacilli.

• Non–spore forming.

• Facultative anaerobes.

• Capable of fermenting glucose “at least”.

• They are oxidase negative

• They reduce nitrates to nitrites

 EPIDEMIOLOGY

❖Enterobacteriaceae inhabit a wide variety of niches, including:

• Human gastrointestinal tract.
• Gastrointestinal tract of other animals.
• Various environmental sites.
❖Mode of transmission:
• By ingestion of contaminated food or water.
• Nosocomial infection.
• Opportunistic colonizer or normal flora (endogenous infection).
• Zoonosis (from animal to human).
• By an insect vector (flea bite).
PATHOGENESIS AND SPECTRUM OF DISEASES

• Endotoxin, a part of the LPS produced by all Enterobacteriaceae, is

responsible for many of the systemic manifestations of infection.
• Endotoxin reactions accompany infections by Enterobacteriaceae, especially
bacteremia.
• Exotoxins, produced by some species and strains, cause diarrhea.
• Adhesins and fimbriae on some species promote adhesion to the
colon, bladder, or other tissues.
• Adhesins on fimbriae prevent bacteria from being washed away and
promote urinary tract infections.
PATHOGENESIS AND SPECTRUM OF DISEASES

• Intracellular growth (Shigella, Salmonella and Yersinia

species and enteroinvasive Escherichia coli) protects
organisms from host defenses.
• Antibiotic resistance develops rapidly and often is encoded
on plasmids, which can be transferred to related bacteria.
• Capsule on Klebsiella and Salmonella species is
antiphagocytic.
• The genes for many of the virulence factors are clustered
and coordinately controlled within pathogenicity islands.
LABORATORY DIAGNOSIS

• No special considerations are required for specimen

collection, transport and processing of the organisms.

• The one exception is Yersinia pestis. This organism is a select

agent. Manipulation of specimens suspected of containing
this organism would generate aerosols and should be handled
using Biosafety Level 3 (BSL-3) conditions.
Gram staining

• Gram staining is Insignificant because of the presence of a

large number of normal gram-negative microbiota.

• Y. pestis resembles a closed safety pin when it is stained

with methylene blue or Wayson stain; this is a key
characteristic for rapid diagnosis of plague.
 CULTIVATION Media of Choice

• Routine laboratory media, such as 5% sheep blood, chocolate, and

MacConkey agars.

• Selective agars, such as Hektoen enteric (HE) agar, xylose-lysine-

deoxycholate (XLD) agar, and Salmonella-Shigella (SS) agar, are
commonly used to cultivate enteric pathogens from
gastrointestinal specimens.
 CULTIVATION Media of Choice

• Cefsulodin-irgasan-novobiocin (CIN) agar is a selective medium

specifically used for the isolation of Y. enterocolitica from
gastrointestinal specimens.

• MacConkey-sorbitol agar (MAC-SOR) is used to differentiate

sorbitol-negative E. coli O157:H7 from other strains of E. coli that
are capable of fermenting this sugar alcohol.
 CULTIVATION Incubation Conditions and Duration

• Most Enterobacteriaceae produce detectable growth in commonly

used broth and agar media within 24 hours of inoculation.

• For isolation, 5% sheep blood and chocolate agars may be

incubated at 35°C in carbon dioxide or ambient air
CULTIVATION Incubation Conditions and Duration

• MacConkey agar and other selective agars (e.g., SS, HE, XLD)
should be incubated only in ambient air.

• Unlike most other Enterobacteriaceae, Y. pestis grows best at 25°

to 30°C. Colonies of Y. pestis are pinpoint at 24 hours but
resemble those of other Enterobacteriaceae after 48 hours.
CULTIVATION Colonial Appearance
 Biochemical test
       
 
I M V C Gelatinase test
E. Coli + ve + ve - ve - ve ❖ All Enterobacteriaceae are Gelatin - ve except
Edwardsiella tarda + ve + ve - ve - ve Proteus Spp. & Serratia odorifera biotype 2
Morganella morganii + ve + ve - ve - ve  
which are Gelatin +ve.
- ve V “ - ve more ” + ve + ve
❖ All Enterobacteriaceae are DNase - ve except
K. pneumoniae
Serratia marcescens, which is DNase + ve.
Enterobacter spp. - ve - ve + ve + ve
Cronobacter sakazakii - ve - ve + ve + ve
Shigella sonnei - ve + ve - ve - ve Proteus spp. VS Morganella morganii &
S. typhi - ve + ve - ve - ve Providencia spp.
➢ Proteus spp. are + ve TSI “Produce H2S”.
S. enteritidis - ve + ve - ve + ve
➢ Morganella morganii & Providencia spp. are -ve
· E. coli (TSI - ve) Vs Edwardsiella tarda (TSI + ve). TSI.
· E. coli (TSI - ve) Vs Edwardsiella tarda (TSI + ve).
· E. coli (TSI - ve) Vs Edwardsiella tarda (TSI + ve). ➢ Those 3 bacteria are Phenylalanine deaminase +
· Shigella sonnei (Non-motile) Vs S. typhi (Motile). ve.
Antimicrobial susceptibility testing and therapy

Extended Spectrum β-Lactamase (ESBL)

• β-lactamases that hydrolyze penicillins and cephalosporins,

including the extended spectrum cephalosporins (cefoxime,
ceftriazone, ceftizoxime, and ceftazidime).
• ESBLs inhibited by clavulanic acid; therefore, this property can be
used as a confirmatory test in the identification process.
Antimicrobial susceptibility testing and therapy

Detection of ESBL
• A chromogenic agar has been developed for the detection of
ESBLs. The agar chrome ID ESBL uses cefpodoxime as a substrate
to increase the recovery and sensitivity of CTX-M type ESBL
isolates.
• ESBLs inhibited by clavulanic acid; therefore, this property can
be used as a confirmatory test in the identification process.
• If isolates test ESBL positive, the results of the antibiotics listed
should reported as resistant.