Salmonella
Introduction
• Genus Salmonella consists of bacilli that
parasitize intestines
• Large number of vertebrate species
• Over 2000 serotypes or species of
Salmonellae
• Potentially pathogenic
Clinically 2 groups
• Typhoidal Salmonellae
– Human
– Enteric fever group
Enteric fever
• Salmonella Typhi
• Salmonella Paratyphi A
• Salmonella Paratyphi B
• Salmonella Paratyphi C
Morphology
• Gram negative bacilli
• 1-3µm x 0.5µm
• Motile by peritrichous flagella
• Noncapsulated
• Nonsporing
Cultural characteristics
• Aerobic and facultative anaerobe
• pH 6-8
• 370C (15-410C)
• Grow on ordinary media
• MacConkey’s agar- non lactose fermenting
Cultural characteristics
MacConkey agar
Cultural characteristics
• Deoxycholate citrate agar(DCA)
• Xylose lysine deoxycholate(XLD)
– Black due to H2S production
Cultural characteristics
• Wilson and Blair bismuth sulphite medium- jet
black
Cultural characteristics
• Selenite F broth
enrichment media
• Tetrathionate broth
Biochemical reactions
• Catalase Positive
• Oxidase negative
• Ferment sugars- glucose, mannitol,
maltose
• Citrate
• H2S production
Glucose Citrate H2Sproduction
[Link] Acid Not utilised Speck of H2S
[Link] A Acid and Not Utilised Not produced
gas
[Link] B Acid and Utilised Abundant H2S
gas
Enteric fever
• Typhoid fever - Salmonella Typhi
• Paratyphoid fever - Salmonella Paratyphi
A, B, C
Typhoid fever
Pathogenesis of typhoid fever
Acquired by
ingestion
Seed liver, Multiply
spleen, bone
Penetrate small marrow, lymph
intestine node Massive
bacteremia
Bloodstream
Phagocytosed via thoracic
by duct Clinical
macrophage, disease
neutrophils Multiply
Reach
Resist Mesenteric back to
intracellular lymph ileum via Typhoid
killing node bile ulcers
Clinical features
• Incubation period 7-14 days
• Onset gradual with headache, malaise, anorexia,
coated tongue, abdominal discomfort
• Stepladder pyrexia
• Soft palpable spleen
• Hepatomegaly
• Rose spots
Complications
• Intestinal perforation
• Haemorrhage
• Circulatory collapse
Paratyphoid fever
• Paratyphoid fever resembles typhoid fever but is
generally milder
Epidemiology
• Source of infection: patient or carrier
• Mode of infection: ingestion of contaminated food
and drink
• Faeces of carriers are important source
• “Mary Mallon”(Typhoid Mary)
Antigenic structure
• 3 important antigens
– Flagellar antigen H
– Somatic antigen O
– Surface antigen Vi
Laboratory diagnosis
• Specimen
• Isolation Blood Culture
Clot culture
Faeces culture
Urine culture
Bone marrow, bile
• Serology
• Demonstration of circulating antigen
• PCR
• Other laboratory tests
Specimen
• Blood, urine, stool, serum, bone marrow
• Specimen depends on duration of illness
Blood culture
• Bacteremia occurs early in disease
Week of illness specimen
1st blood
2nd serum
3rd stool
4th urine
Blood culture
Method
• 5-10ml in 50-100ml of 0.5% bile broth
• Incubate at 37oC
Subcultured on MacConkey’s agar
Pale nonlactose negative
fermenting colonies
repeat subculture for
10 days
Biochemical reactions
Other specimens
• Clot culture:
– 5ml of blood is withdrawn and allowed to clot
– Serum is separated and clot is broken, added to
bile broth
• Faeces: enrichment media and selective media is
used.
Morphology
• Gram negative bacilli
• 1-3µm x 0.5µm
• Motile by peritrichous flagella
• Noncapsulated
• Nonsporing
Cultural characteristics
• Aerobic and facultative anaerobe
• pH 6-8
• 370C (15-410C)
• Grow on ordinary media
• Large, circular, low convex, smooth, translucent
• MacConkey’s agar- non lactose fermenting
Cultural characteristics
MacConkey agar
Cultural characteristics
• Deoxycholate citrate agar(DCA)
• Xylose lysine deoxycholate(XLD)
– Black due to H2S production
Cultural characteristics
• Wilson and Blair bismuth sulphite medium- jet
black
Cultural characteristics
• Selenite F broth
enrichment media
• Tetrathionate broth
Biochemical reactions
• Catalase positive
• Oxidase negative
• Ferment sugars- glucose, mannitol, maltose
• Citrate
• H2S production
Glucose Citrate H2Sproduction
[Link] Acid Not utilised Speck of H2S
[Link] A Acid and Not Utilised Not produced
gas
[Link] B Acid and Utilised Abundant H2S
gas
Serotyping by slide agglutination with O
and H antisera
Serology – Widal test
• Tube agglutination test
• For measurement of O and H antibodies for
typhoid and paratyphoid bacilli in the serum
• 2 types of tubes
– Felix tube- round bottom for O agglutination
– Dreyer’s tube- conical bottom for H
agglutination
Widal test
• Equal volumes of serial dilutions of serum(1/20-
1/640)
• Mixed with O and H antigens of [Link] and H
antigens of S. Paratyphi A and B
• Incubate at 370C overnight
Widal test
• Aggutination titres are read
– H suspensions agglutinate rapidly – large, loose,
fluffy clumps
– O antigen suspensions- compact, chalky,
granular clumps
Widal test
Widal test
Widal test
• Agglutination titre depends on stage of disease
• Demonstration of rise in titre
• Results of single test – interpret with caution
– Antibodies may be present on account of prior
disease, infection, immunisation
– H antibodies persist longer than O
– Anamnestic response
– Cases treated early with antibiotics may show
poor titre
PCR
• Sensitive
• Not widely available
Demonstration of antigen
• In early phase of disease
• In blood and urine
• Staphylococcal coagglutination test
Other laboratory tests
• WBC count- leucopenia with relative
lymphocytosis
Diagnosis of carriers
• Epidemiological and public health purpose
Methods
• Isolation from faeces and bile
• Vi antibody detection
• Sewer swab technique
• Filtration of sewage through millipore membrane
and culture on Wilson and Blair media
Prophylaxis
• General measures- improvement in sanitation ,
protected water supply
• Specific measures- vaccines
Vaccines
• TAB vaccine
– Heat killed vaccine
– [Link] 1000 million, [Link] A and B 750
million
– 0.5ml subcutaneous
– 2 doses 4-6 weeks apart
Vaccines
• Typhoral vaccine
– Live oral vaccine
– Stable mutant of [Link] strain Ty21a, lacking
enzyme UDP-galactose-4-epimerase
– Enteric coated capsule
– 109 bacilli
– 1 hour before food on days 1,3,5
Vaccines
• Vi vaccine
– Injectable vaccine (Typhim-Vi)
– Purified polysaccharide Vi antigen of [Link]
strain Ty2
– Single subcutaneous or im dose
Treatment
• Chloramphenicol
• Ciprofloxacin
• Ceftriaxone
• Ampicillin
• Amoxycillin
• Cotrimoxazole
• Problem of drug resistance
