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Practical

Diagnosis of Salmonellosis

About 10 gm of each sample will be collected in separate plastic bags. The samples will be pre-enriched
in buffer peptone water, which were further enriched in selenite broth incubating at 42°C for 18-24
hours. The broth cultures will be transferred to the selected media of deoxycholate agar. After
incubation, the suspected colonies will be subcultured on McConkey agar. The pure colonies will be
maintained in the nutrient agar slant for further identification.
The following tests will be conducted for further identification:
1. Slide agglutination test with polyvalent Salmonella O sera
2. Biochemical test
3. Sugar test

Samples (2 gm)

Pre-enriched in buffer peptone

Incubation

Selenite broth

Incubation

Selective media (Xylose lysine deoxycholate agar)/BGA

Incubation

Mac Conkey agar


 Slide examination
 Catalase
 Oxidase
 TSI ( Triple sugar iron) -+ve
 MRVP(Methyl red voges proskauer)
 Oxidase fermentation media
 Citrate media
Table. Pure culture isolation of Salmonella spp using 10 sugar fermentation tests

Sugars S. enteritidis S. cholerasuis S. typhi S. Kaufmannii S. salamae S. arizonae S.. pullorum

Dulcitol + - - + + - -
Xylose + + + + + + -
Maltose + + + + + + -
Lactose + - - - - - -
Arabinose + + + + + + +
Sucrose - - - - - - -
Trehalose + + + + + + +
Adinosine - - - - - - -
Mannitol + + + + + + +
Inositol - - - - - - -