Salmonella Spp.

HIRENDRA D. TADVI
UNIT : CLINICAL BACTERIOLOGY
INTRODUCTION
 The genus Salmonella consist of bacilli. The most important member
of the genus is Salmonella typhi, the causative agent of typhoid fever.
 The typhoid bacilli was first observed by Eberth(1880) in the
mecentric nodes and spleen of fatal case of fever and was isolated by
Gaffky(1884).The first series to be isolated from animals and human
beings- The genus Salmonellae.
 Salmonellae currently comprise above 2000 serotype or species, all of
them potentially pathogenic.
 The Genus Salmonella are killed at 55°C in 1 hour or at 60°C in 15
minutes.
 Boiling, chlorination of water, pasteurization of milk destroys the
organism.
 They can survive for weeks in polluted water and soils and for months
in ice.
 For practical purpose, they may be divided into two groups
i. Typhoidal : Enteric fever group
• Primary human parasite.
ii. Non typhoidal : Food poisoning group
• Animal parasite which can also infect human.
MORPHOLOGY

 Salmonella is
 Gram-negative, rod-shaped
 1-3 μm X 0.5 μm in size
 Non-acid fast
 Non-capsulated and non-sporing
 Facultative anaerobe in family Enterobacteriaceae
 Motile, Non lactose fermenting
CULTURAL CHARACTERISTICS

 Salmonellae are aerobic and facultatively anaerobic, growing

readily on simple media over a range of pH 6-8 and temperature 15-
41°C (optimum 37°C ).
 Colonies: Large ,2-3 diameter, circular, low convex and smooth.
 More translucent than coliform colonies.
 Selective media for subculture from blood culture bottle:
 MacConkey: Colonies are colorless due to the absence of lactose
fermentation.
CULTURAL CHARACTERISTICS
 Deoxycholate citrate agar : Colonies are colorless due to the
absence of lactose fermentation.
 Xylose lysine Deolycholate agar : Colonies are colorless.
 Wilson Blair : Bismuth sulphite medium, jet black colonies with a
metallic sheen are formed due to production of H2S produce green
colonies.
 Tellurite blood agar
 Enrichment broth for culture of stool/urine :
 Selenite F and tetrathionate broth
BIOCEHMICAL CHARACTERISTICS

 Fermentation tests:
 Salmonellae ferment glucose, mannitol and maltose, forming acid
and gas.
 An important exception is S. typhi which is anaerogenic.
 Lactose, sucrose and salicin are not fermented.
 Decarboxylase tests
 Salmonella decarboxylase the amino-acids: lysine, ornithine and
arginine but not glutamic acid.
BIOCEHMICAL CHARACTERISTICS
 Other Biochemical tests:
 Most Salmonella have following characteristics:
 Indole -ve, MR +ve, VP -ve, Citrate utilize (Except by S. Typhi and
S. Paratyphi A).[IMViC -+-+].
 Malonate not utilized, gluconate not utilized, urease -ve, H2S
produced in Ferrous chloride gelatin medium except by S. Paratyphi
A.
 No growth in KCN medium, gelatin not liquify.
Pathogenesis of Salmonella
 Salmonellosis = Generic term for disease
 Clinical syndromes caused in human beings:
 Enteritis (Acute Gastroenteritis):Food Poisoning
• Zoonotic disease.
• Caused by Salmonella except S. typhi. Most common by S. Typhimurium.
• High infectious dose (108 CFU).
• 6-48h incubation period.
• Nausea, vomiting, nonbloody diarrhea, fever, cramps, myalgia and headache common.
• Source : Poultry , Meat , Milk and its products,esp.Eggs and its products (as the
bacteria can enter through the shell if eggs are contaminated and grow
inside),uncooked vegetables.
 Enteric fever :
• S. typhi causes typhoid fever and S. paratyphi A, B and C cause milder
form of enteric fever called paratyphoid fever.
• Carriers: 1.Convalescent carrier
2.Temporary carrier
3.Chronic carrier
• About 2-4 % of patient chronic carrier.
• Most common in female(older age group >40y).
• Fever, headache ,anorexia ,coated tounge and abdomen discomfort, rose
spot.
• Infectious dose large = 106 CFU.
 Fecal-oral route of transmission:
 Person-to-person spread by chronic carrier
 Fecally-contaminated food or water
 Food handlers contaminate food
• 10-14 day incubation period
 Salmonella septicemia :
• Causes by S. Choleraesus.
• During this stage the bacilli are entered to the gall bladder, spleen,
liver bone marrow, lungs and kidney and the further multiplication
occurs.
• After multiplication it enters to blood and causes secondary
multiplication.
• During this time they produce toxins and causes fever and other
symptoms.
• Isolated from blood or pus.
Pathogenesis
 Lab diagnosis
 For Lab diagnosis, specimen & diagnostic tests according to duration
of fever:
• Blood for Culture
• Urine culture
• Feces culture
• Serum (WIDAL)
 For blood culture
 Salmonella is present in the blood of more than 80% of patients with typhoid
fever. The overall volume of blood cultured is critical to increase yield
(isolation rate) of the causative pathogen. Reducing the blood volume reduces
the sensitivity of the blood culture.
 5-10 ml blood collected and inoculate in culture bottle.
 Subculture it into culture media.
 Subculture plates should be incubated at 37°C for 18-24 hours in an
aerobic incubator. If growth is observed in the culture plates, colony
morphology should be noted and biochemical tests performed to identify
the isolate.
 Alternative of blood culture is clot culture, here 5 ml of blood collected
into sterile pipette and allow it to clot. The serum is pipette and used for
widal test.
 Urine culture:
 Salmonella are shed in urine irregularly hence urine culture is less useful
then blood culture.
 In this only 25 % of the cases has chances of positive.
 Faces culture:
 It is as valuable as blood culture.
 Fecal culture is particularly valuable in patients on antibiotics as the drug
does not eliminate the bacilli from the gut as rapidly as it does from the
blood.
 Other material for culture: Bone marrow is valuable in most cases.Other
material which is pus from supportive lesions ,CSF and sputum. At autopsy
cultures may be obtained from the gall bladder ,live ,spleen and mesenteric
lymph nodes.
 Widal Test
 This test has only moderate sensitivity and specificity.
 It can be negative in up to 30% of culture-proven cases of typhoid
fever.
 Widal test measures agglutinating antibody levels against O and H
antigens.
 As S.typhi shares O and H antigens with other Salmonella serotypes
and has cross-reacting epitopes with other Enterobacteriaceae, false
positives widal test results may occur in other clinical conditions
such as malaria, typhus fever, bacteremia caused by other organisms
as well.
PROPHYLAXIS

 General measure:
 Improvement in sanitation.
 Provision of protected water.
 Vaccine
 TAB vaccine :
 Heat killed typhoid bacillus vaccine.
 Contain S. typhi 1000 million and S. paratyphi A & B 750 million
each/ml.
 Oral Ty21-A(Typhoral) :
 Live attenuated, Salmonella typhi vaccine.
 Consist of stable mutant of S. typhi strain TY21-A.
 Vi vaccine :
 Capsular polysaccharide vaccine.
 Contain S. typhi strain Ty2.