Culture media for Salmonella typhi and paratyphi

Salmonella is a member of the family Enterobacteriaceae. It is a Gram-negative bacilli, motile and non-lactose fermenter. The genus can be divided into two species (S. enterica and S. bongori), based on their phenotypic profile.

It causes acute gastroenteritis and when Salmonella infections become invasive, they can affect the bloodstream (bacteremia), bone (osteomyelitis), joint (septic arthritis), brain, or nervous system (meningitis). Salmonella bacteria are classified as either “typhoidal” or “nontyphoidal,” based on their serotype.

  • Typhoidal Salmonella refers to the specific Salmonella serotypes which cause typhoid fever or paratyphoid fever, including typhi, paratyphi A, paratyphi B, and paratyphi C.
  • Nontyphoidal Salmonella refers to all other Salmonella serotypes.

For the recovery of Salmonella from clinical specimens, three general types of media are available.

  1. Non-selective media for primary isolation (blood agar)
  2. Selective or differential agar (e.g. MacConkey agar, hektoen enteric agar); and
  3. Enrichment broths (e.g. selenite broth)

Commonly used culture media for isolation of Salmonella and their colony characteristics:

  1. Bismuth sulfite agar: Salmonellae produce black colonies.
  2. Blood AgarS. typhi and S. paratyphi usually produce non-hemolytic smooth white
    colonies.
  3. MacConkey Agar: Non-lactose fermenting smooth colonies i.e. pale colonies
  4. Deoxycholate Citrate Agar (DCA): Salmonella appear as pale colonies.
Colonies of Salmonella typhi in DCA Agar. Source: bacteriainphotos.com

5. Xylose Lysine Deoxycholate (XLD) Agar: Salmonella appear as pink (lactose non-fermenting colonies) with a black center (due to H2S production)

6. Salmonella-Shigella (SS) Agar:  Used for selective recovery of Salmonella from stool culture (contains five times the concentration of bile salts compared with MacConeky Agar). Salmonellae usually produce lactose non-fermenting colonies with black centres (except S. paratyphi A, whose colonies do not have black centres).

7. Hektoen Enteric (HE) Agar: On hektoen enteric agar, salmonellae produce transparent green colonies with black centres (except S. paratyphi A, whose colonies do not have black centres).

8. Eosin Methylene Blue (EMB) Agar: Non-lactose fermenters, including Salmonella, Shigella and Proteus forms colorless or transparent colonies.

9. Wilson Blair media:

  1. S.Typhi forms large black colonies with a metallic sheen.
  2. S.Paratyphi A produces green colonies due to the absence of H2S production.

10. Selenite broth or Tetrathionate broth: For the enrichment of Salmonellae present in feces. Feces is incubated for 12-18 hours before subculture into plates.

11. Trypticase soy broth or brain heart infusion broth: Non-selective broth for primary blood culture.

Biochemical Tests

Commonly used Biochemical tests for the identification of Salmonella and their results are as follows;

Biochemical test for Salmonella
Salmonella typhi in TSI agar or KIA agar
Salmonella typhi in TSI agar or KIA agar
Biochemical TestsResult
Catalase testPositive
Oxidase testNegative
Indole testNegative
Voges-Proskauer (VP) test (media: MR/VP broth)Negative
Methyl-red (MR) testPositive
Citrate Utilization test (media: Simmon’s citrate agar)Positive 
Urea hydrolysis test (urease test in urea medium)Negative
Nitrate recduction testNegative
Triple sugar iron agar (TSI) testAlk/Acid, H2S
Arginine-dihydrolase testVariable
Lysine-decarboxylase testPositive
Ornithine-decarboxylase testPositive
Sucrose/salicinUsually not fermented

References and Further Reading

  1. Ruiz, J., Núñez, M. L., Lorente, I., Pérez, J., Simarro, E., & Gómez, J. (1996). Performance of six culture media for isolation of Salmonella species from stool samples. European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology, 15(12), 922–926. https://doi.org/10.1007/BF01690509
  2. Uyttendaele, M., Bagamboula, C. F., De Smet, E., Van Wilder, S., & Debevere, J. (2001). Evaluation of culture media for enrichment and isolation of Shigella sonnei and S. flexneri. International journal of food microbiology, 70(3), 255–265. https://doi.org/10.1016/s0168-1605(01)00549-9 
  3. Dekker, J. P., & Frank, K. M. (2015). Salmonella, Shigella, and yersinia. Clinics in laboratory medicine, 35(2), 225–246. https://doi.org/10.1016/j.cll.2015.02.002

Acharya Tankeshwar

Hello, thank you for visiting my blog. I am Tankeshwar Acharya. Blogging is my passion. As an asst. professor, I am teaching microbiology and immunology to medical and nursing students at PAHS, Nepal. I have been working as a microbiologist at Patan hospital for more than 10 years.

11 thoughts on “Culture media for Salmonella typhi and paratyphi

  1. one of the best student learning and review webs by far…..SPECIFIC to the point and NO JUNK thrown around to fill in space !!!

  2. If we incubate any bacterial pure culture like E.coli,B.cereus,P.aeruginosa more than 16 to 18 hours. What is the impact of it on the survival of the culture.

  3. Sor… I love so much to do culture to the stool sample fo s. Typhi and other diseases, urine fo the chronic U. T. I and H. V. S…, so can help me to get clear whole procedure,result and how to treat the infection of a such diseases.
    Diagram will be best for the each test

  4. Hi,
    Can you please let me know for standard environmental swab samples in poultry production should enrichment broth be used or dry agar according to ISO 6579:2002 amended 2007.
    It would seem the method involving enrichment broth prior to the amended standard would likely find salmonella and is not appropriate and is why the amended 2007 version as I understand it does not use enrichment.
    I would be grateful if you could clarify what is the correct test method. Thank you.
    Noel

  5. HI
    I like the sit and do culture of stool for confirmation of salmonella typi and paratypi
    so can u classify for weather the use of rapid salmonella typi test are correct in stool diagnosis
    can u make the interpretation of biochemical test for the identification of salmonella typi easy

  6. can you please write about molecular techniques for identifying bacteria, fungi or maybe specifically for diseases such as salmanellosis

  7. Tankeshwar sir, I’ve a question. I have acquired blood samples from the patients suffering from typhoid fever and done VIDAL test to confirm the presence of S. typhi, I want to culture the microbe on MacConkey Agar from blood. I am a bit confused as how to do it because its my first time doing so. Your help is highly appreciated.

    1. Muhmmad, you have to first inoculate the blood sample in Broth (Brain Heart Infusion Broth or Bactec Blood Culture System). Find more about Blood Culture here. Sub-culture is done in solid culture medium (Blood Agar, Chocolate Agar and MacConkey Agar) only if growth is noted in the broth medium.

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