Enterobacteriaceae

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

Aerobic or facultative anaerobic

Motile by peritrichate flagella

Grow on ordinary media (non fastidious)

Ferments glucose with acid & gas or only acid

Catalase + ve & oxidase -ve

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Classification of Enterobacteriaceae

 Based on lactose fermentation – oldest method :

• Lactose fermenters e.g. Escherichia, Klebsiella
• Late lactose fermenters e.g. Shigella sonnei
• Non lactose fermenters e.g Salmonella, Shigella
- Commensal intestinal bacteria: LF
- Intestinal pathogens: NLF

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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.

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Antigenic Structure of Gram –ve Bacteria

 Three antigens – serotyping

of E.coli
• H – flagellar antigen
• O – somatic antigen
• K – capsular antigen

Majority do not possess K Ag.

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Virulence Factors

- Two types of virulence factors: Surface Ags & Toxins

2. Surface Antigens
 LPS surface O Ag – endotoxic activity
 Envelope or K Ag – protects against phagocytosis
 Fimbriae – colonisation, found in strains causing diarrhoea and
urinary tract infections

 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

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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)

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 Heat Labile Toxin (LT)

Escherichia coli /
Vibrio cholerae
Gut lumen

Intestinal
epithelial cell

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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

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 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

Gram stain Gram negative bacteria

(1bacteria / oil field is significant)

Urine Culture To know significant bacteriuria

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 Lab Diagnosis of E. coli UTI

Significant > 105 organism / ml of MSU

bacteriuria

Culture BA / MAC : LF (flat)

CLED medium

Identification tests

I M Vi C test: + + - -
TSI agar Acid, no gas

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Diarrheagenic E.coli

 Enteropathogenic E. coli (EPEC) – infantile diarrhea, nontoxigenic

 Enterotoxigenic E. coli (ETEC) – traveller’s diarrhea, resembles cholera

 Enteroinvasive E. coli (EIEC) – bloody diarrhea (blood, mucus &

leucocytes with stool)

 Enterohemorrhagic E. coli (EHEC) or Verocytotoxigenic E. coli

(VTEC):- O157:H7 serotype (food poisoning)

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

persistent diarrhea in children

 Diffusely adherent E. coli (DAEC)

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Epidemiology & Treatment

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

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

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 Klebsiella

Normal gut flora in the intestine

Gram negative bacilli (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

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Pathogenicity of Klebsiella pneumoniae
 Pulmonary infections - Lobar Pneumonia :
• 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.

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Lab Diagnosis - Klebsiella

Specimens Urine, sputum, nasal

secretions / swab, blood

Culture BA / MAC : LF (mucoid)

Biochemical TSI agar Acid with gas

tests
Urease Positive

Citrate Positive

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 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

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 Shigella

 Bacillary Dysentery – frequent passage of blood

stained, mucopurulent stools.

 Classification – 4 species : biochemical & serological

characteristics.
- Sh.dysenteriae
- Sh.flexneri Non Lactose Fermenter
- Sh.boydii
- Sh.sonnei - Late Lactose Fermenter

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 Shigella species- Mannitol fermentation
Mannitol

Non Fermentation Fermentation

S. dysenteriae - 12 S. flexneri- 6

S. boydii - 18

S. sonnei
(Late lactose fermenter)

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 Characteristics

 Gram–ve, nonmotile bacilli.

 Enrichment broth – Selenite F, Gram-ve broth.

 Selective media – Deoxycholate agar(DCA)

- Salmonella-Shigella agar
- XLD (Xylose Lysine deoxycholate)

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Epidemiology & Clinical Syndromes

 Incubation period: 1-7 days, usually 48 hrs

 Low Infectious dose: 10-100 bacilli
 Feco-oral transmission
 Common in pediatric age group (1-10 years) – leading cause of
infantile diarrhea.
 Sh.dysenteriae type I : most serious form of dysentery.
 Shigellosis : whole spectrum of disease caused by Shigella.
 Complication: Hemolytic Uremic Syndrome

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 Pathogenesis Two-stage disease

Early stage
Watery diarrhea attributed to the enterotoxin activity of Shiga toxin in
the small intestine

Second stage
Dysentery due to adherence and tissue invasion of large intestine
(cytotoxic activity of Shiga toxin)

Fever attributed to neurotoxic activity of toxin

Shiga toxin Enterotoxic, neurotoxic and cytotoxic

Similar to Shiga-like toxin of Enterohemorrhagic E. coli (EHEC)

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Laboratory Diagnosis

 Specimen: fresh feces – mucus flakes (buffered glycerol saline –

transport medium)

 NLF colonies .
 Slide agglutination with polyvalent & monovalent sera.

Treatment
 Oral rehydration
 Antibiotics for severe & toxic cases – Nalidixic acid or Norfloxacin.

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 Salmonella

 Gut of domestic animals & poultry.

 Divided into 2 groups :
• Enteric fever group – typhoid & paratyphoid bacilli.
• Food poisoning group – usually animal parasites, producing
gastroenteritis, septicemia or localized infections.

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 Morphological & Cultural characteristics

 Motile, gram negative bacilli

 Colorless (NLF) on MacConkey &

DCA.

 Enrichment broth- Selenite F,

Tetrathionate broth

 Selective media – Wilson & Blair (jet

black colonies due to H2S), XLD, SS
agar.

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 Pathogenesis

Source Carriers Cases Poultry, dairy

Transmission Ingestion of contaminated water or food

High infectious dose (108 CFU)

IP 7-14 days

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 Inf ecti on pattern of Sal monel la
Salmonella are ingested in
contaminated food or water

Organisms reach the Enteritis

terminal ileum
Organisms invade the gut wall
& cause ulcertion, perforation
& hemorrhage
Organisms spread to intestinal
lymphatics & are phagocytosed by
macrophages

Organisms disseminate to Enteric fever or

bones, kidneys, lungs,liver, typhoid fever
brain & blood

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 Pathogenicity

 Enteric fever – Typhoid & paratyphoid fever.

- Clinical features: nausea, vomiting, fever, bradycardia, toxemia,
splenomegaly, hepatomegaly, diarhoea alternating with constipation.

 Septicemia with or without local suppurative lesions.

 Gastroenteritis

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 Lab diagnosis of Enteric fever
Specimens Blood, Bone marrow, urine, stool, pus, CSF

Blood culture BHI broth

1st week

2nd week Antibody detection Widal test

(serum)

3rd week Urine culture

4th week Stool culture Use selective & enrichment

medium

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 Serology - Widal Test
Tube agglutination test To detect antibodies in patient serum

Test is performed after 2 wks To diagnose Enteric / typhoid fever

Antigens used TO O antigen of S typhi

TH H antigen of S typhi
AH H antigen of paratyphi A

BH H antigen of paratyphi B

CH H antigen of paratyphi C

O is group specific Enteric fever

H is species specific Typhoid or paratyphoid

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 Carriers
Food handlers & Cooks
Repeated stool cultures
Vi agglutinins indicates carrier status

Vaccin es
TAB
Typhoral
Typhim

Tr eatm ent
Ciprofloxacin

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 Vibrio
 V.cholerae, the causative agent for cholera.
 Water contaminated with feces of patients & carriers.

Morphology
 Gram-ve slender bacilli, comma shaped.
 Actively motile by a long polar flagellum
- Darting motility.

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Cultural characteristics

 Aerobe, alkaline pH (> 8).

 Transport media – Cary Blair media,

VR (Venkataraman-
Ramakrishnan)media

 Enrichment media – Alkaline Peptone

Water, Monsur’s taurocholate tellurite
PW.

 Selective media – Alk. bile salt agar,

TCBS agar.

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 Biochemicals
 Oxidase +ve

 Ferments sucrose,
late lactose fermenter.

 ‘String test’ +ve

( with 0.5% sodium deoxycholate ).

 ‘Cholera red’ reaction – conc. Sulphuric acid to peptone

water culture.

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Classification

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 Pathogenicity

 Cholera – acute diarrhoeal disease, passage of large

volumes of rice watery stools, dehydration, hypovolemic
shock & metabolic acidosis.

 ID – 1011 org/ ml

 Cholera toxin (CT)– endotoxin

 Toxin coregulated pilus (TCP)

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Laboratory Diagnosis

 Stool examination – before giving antibiotics.

 Motility
 Culture – Non selective (BSA, Mac), Selective (TCBS).
BA – Hemodigestion.
 Slide agglutination
 Phage typing – NICED, Calcutta (INDIA).

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 Prophylaxis & Treatment

 Oral vaccines – immunity lasts for 6- 12 months.

 ORF

 Antibiotic therapy of secondary importance.

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