Enterobacteriaceae

Dr Ekta Chourasia
Microbiology
General Features of Enterobacteria
Present in large intestine
Gram negative bacteria

Aerobic or facultative anaerobic

Motile by peritrichate flagella or non motile

Grow on ordinary media (non fastidious)

Ferments glucose with acid & gas or only acid

Reduce nitrates to nitrites

Catalase + ve & oxidase -ve

Dr Ekta
Classification of Enterobacteriaceae

 Based on lactose fermentation – oldest method :

1. Lactose fermenters e.g. Escherichia, Klebsiella.
2. Late lactose fermenters e.g. Shigella sonnei
3. Non lactose fermenters e.g Salmonella, Shigella

- Commensal intestinal bacteria: LF

- Intestinal pathogens: NLF

Dr Ekta
Classification of Enterobacteriaceae

 Modern taxonomy – group together bacteria that

possess:
1. Common morphological and biochemical properties
2. Similar DNA base compositions

 Family – Tribe / Group - Genera

Dr Ekta
 Enterobacteriaceae (Tribes & Genera)
CDC 1989

Tribe 1 Eshcherichieae Tribe 5 Klebsielleae Klebsiella

Escherichia Serratia Enterobacter

Shigella Hafnia Pantoea

Tribe 2 Edwardsielleae Tribe 6 Proteeae Proteus

Edwardsiella Providentia Morganella

Tribe 3 Salmonelleae Tribe 7 Yersinieae

Salmonella Yersinia

Tribe 4 Citrobactereae Tribe 8 Erwinieae

Erwinia
Citrobacter

Dr Ekta
Escherichia coli

 Named after Escherich, first to describe colon bacillus

 Normal flora of the human & animal intestine.
 Remains viable in the feces for few days.
 Detection of E. coli in the drinking water – indicates
recent pollution with human or animal feces.

Dr Ekta
Antigenic Structure of Gram –ve Bacilli

 Three antigens – serotyping

of E.coli
1. H – flagellar antigen
2. O – somatic antigen
3. K – capsular antigen

Majority do not possess K Ag.

Dr Ekta
Virulence Factors

- Two types of virulence factors: Surface Ags & Toxins

1. Surface Antigens
- LPS surface O Ag – endotoxic activity, protects from
phagocytosis and bactericidal effects of complement

- Envelope or K Ag – protects against phagocytosis and

antibacterial factors inserum

- Fimbriae – colonisation factors, found in strains causing

diarrhoea and urinary tract infections

Dr Ekta
Virulence Factors

2. Toxins (Exotoxins) – two types

- Enterotoxins – pathogenesis of diarrhoea
- 3 types : LT (heat labile toxin),
ST (heat stable toxin) &
VT (verocytotoxin or shiga- like toxin)

- Hemolysins – may be nephrotoxic

Dr Ekta
Heat Labile Toxin (LT)

 Resembles cholera toxin in its

structure, function and mode of
action
 Complex of polypeptide
subunits.
 LT: one subunit of A
(action- enzymic),
five subunits of B (binding)

Dr Ekta
 Heat Labile Toxin (LT)

Escherichia
Escherichiacoli
coli/ /
Vibrio
Vibriocholerae
cholerae
Gut lumen

Intestinal
epithelial cell

Dr Ekta
E.coli toxins

Heat Labile Toxin (LT) Heat Stable Toxin (ST)

Activates Adenyl cyclase Activates Guanyl cyclase

increased production of cAMP Increased production of cGMP

Increased secretion of Na, Cl and water Inhibition of ionic uptake in intestinal cells
from the cell

Osmotic loss of water from cells

Dr Ekta
Pathogenicity/ Clinical Infections
1. Urinary tract infection
2. Diarrhoea
3. Pyogenic infections
- Wound infection, especially after surgery of lower intestinal tract.
- Peritonitis.
- Biliary tract infection.
- Neonatal meningitis.
4. Septicemia – can lead to fatal conditions like
- Septic shock
- Systemic Inflammatory Response Syndrome

Dr Ekta
Lab Diagnosis of UTI

Specimens Urine Mid stream urine (MSU)

Catheter specimen urine (CSU)
Supra pubic aspiration (SPA)

Microscopy
Wet mount Pus cells / hpf
Bacteria / crystals/ casts

Gram stain Gram negative bacteria

(1bacteria / oil field is significant)

Urine Culture Kass semi-qauntative method

Standard loop technique
To know significant bacteriuria

Dr Ekta
 Lab Diagnosis of E. coli UTI

Significant > 105 organism / ml of MSU

bacteriuria

Culture BA / MAC : LF (flat)

Identification tests

I M Vi C test: + + - -

TSI agar Acid, no gas

AST

Dr Ekta
Diarrheagenic E. coli

 Enteropathogenic E.coli (EPEC)

 Enterotoxigenic E.coli (ETEC)

 Enteroinvasive E.coli (EIEC)

 Enterohemorrhagic E.coli (EHEC) or Verotoxigenic E.coli (VTEC)

 Enteroaggregative E.coli (EAEC) : “stacked brick” appearance.

 Diffusely adherent E.coli (DAEC)

Dr Ekta
Enteropathogenic E. coli (EPEC)

 Infantile diarrhea
 Institutional outbreaks
 Noninvasive, nontoxigenic
 Pathogenesis – adhesion via fimbria, disruption of brush border
microvilli
 Clinical features – fever, diarrhea, vomiting, nausea, non bloody
stools
 Lab Diagnosis – testing colonies grown on BA/ MA with EPEC
O antisera

Dr Ekta
Enterotoxigenic E. coli (ETEC)

 Traveller’s diarrhea
 Resembles cholera
 Noninvasive, toxigenic
 Pathogenesis – production of plasmid coded toxins (LT/ ST)
 Clinical features - Diarrhea, vomiting and abdominal pain
 Lab Diagnosis – demonstration of enterotoxin by in vitro or in
vivo methods, detection of LT/ St by gene probes

Dr Ekta
Enteroinvasive E. coli (EIEC)
 Bloody diarrhea (dysentery), resembles Shigella
dysentery
 Passage of blood, mucus & leucocytes in stool
 Pathogenesis - Invades epithelial cells by endocytosis
and can spread laterally to adjacent cells, causes tissue
destruction, necrosis and ulceration.
 Lab Diagnosis:
1. Sereny test - instillation of suspension of freshly isolated EIEC
or Shigella in the eyes of guinea pig – mucopurulent
conjunctivitis and severe keratitis
2. Penetration of HeLa or Hep2 cells in tissue culture

Dr Ekta
Dr Ekta
Enterohemorrhagic E. coli (EHEC)
 Produces verocytotoxin (VT), a shiga-like toxin (SLT); hence also
known as Verocytotoxigenic E.coli (VTEC)
 Pathogenesis – EHEC attaches to the colonic mucosa and
releases VT. VT targets vascular endothelial cells, inhibits protein
synthesis - cytotoxicity
 Clinical features - Mild diarrhea (bloody) to fatal complications
(esp. in young children and elderly):
1. Hemorrhagic colitis – destruction of mucosa followed by hemorrhage.
2. Hemolytic Uremic syndrome – triad of acute renal failure, hemolytic
anemia and thrombocytopenia.
 Serotype O157: H7 is most commonly involved.
 Outbreaks of food poisonings (fast foods, contaminated
hamburgers)

Dr Ekta
Enterohemorrhagic E. coli (EHEC)

 Lab Diagnosis:
1. Demonstration of bacilli or VT in feces or in culture
2. Sorbitol MacConkey agar for O157:H7 – does not ferment sorbitol
unlike other E. coli
3. Cytotoxic effects on Vero or HeLa cells
4. DNA probes to detect toxins

Dr Ekta
Enteroaggregative E. coli (EAEC)

 Persistent diarrhea in children in developing countries.

 Aggregate to give a “Stacked brick appearance” on Hep2 cells

or glass (due to fimbria)

 Pathogenesis – shortening of villi, mucus biofilm, heat stable

cytotoxin (hemorrhagic necrosis and edema)

Dr Ekta
Epidemiology & Treatment

Epidemiology
 EPEC & ETEC - most important causes of diarrhea globally
 EHEC – in developed countries.

Treatment
 Based on symptoms:
1. Primary treatment – fluid replacement
2. Secondary treatment – antibiotics in severe cases with systemic involvement

Dr Ekta
 Klebsiella

Normal gut flora in the intestine

Gram negative coccobacilli (short &
plump)
Capsulated, non-motile, Mucoid LF colonies on MAC

Species
K. pneumoniae Pneumonia, Urinary tract infections
K. oxytoca
K. ozaenae Atrophic rhinits
K. rhinoscleromatis Rhinoscleroma

Dr Ekta
Pathogenicity of Klebsiella pneumoniae
 Pulmonary infections - Pneumonia (lobar):
1. High fatality
2. In middle aged or older persons with medical problems like DM,
alcoholism, chronic bronchopulmonary disease
3. Extensive necrosis & hemorrhage resulting in thick, mucoid, brick
red sputum “currant jelly like”
 Extrapulmonary infections –
1. Meningitis & enteritis in infants
2. UTI
3. Septicemia
 An important cause of nosocomial infections.

Dr Ekta
Lab Diagnosis - Klebsiella

Specimens Urine, sputum, nasal secretions / swab,

blood

Culture BA / MAC : LF (mucoid)

Identification tests

I M Vi C test: - - + +

TSI agar Acid with gas

Urease Positive

Dr Ekta
 Proteus
Normal gut flora in the intestine

Gram negative bacilli, pleomorphic

Motile, Non lactose fermenter NLF on MAC

Swarms on BA, Urease +, H2S +

Species P. mirabilis P. vulgaris

UTI Pneumonia Wound infections

Urease converts urea to NH4 & CO2 causing alkalinization of

urine leading to renal calculi (stones)

Proteus antigens are used in the Weil - Felix test to

diagnose Rickettsial diseases

Dr Ekta
Lab Diagnosis - Proteus
Specimens Urine, sputum, wound swab

Culture BA: swarming

MAC : NLF, fishy/ seminal smell

Identification tests
Indole: PM - / PV + Urease +
TSI agar K / A (H2S)

Dr Ekta
Enterobacter, Serratia, Citrobacter

 Moist environments in hospitals – common reservoirs.

 Pathogenicity –
- UTI,
- Wound & respiratory infections in hospitalized patients,
- Outbreaks in ICUs, burn units & other special units

Dr Ekta