Bacterial Pathogen Isolation from Blood and Stool

Laboratory methods for culturing bacterial pathogens from blood have been described previously [28 (link)]. Briefly, blood (7–10 ml for adults and 1–3 ml for children) was inoculated into BACTEC culture bottles (Becton, Dickinson and company, Sparks, MD, USA) and incubated in an automated BACTEC 9050 at 35°C for 1–5 days. A Gram-stained smear was prepared from any bottle with growth, and the broth sub-cultured onto standard enriched culture media for further identification. Stool specimens were processed according to standard protocols described elsewhere [33 (link)]. Colonies of Salmonella were identified and confirmed using an API 20E system (Appareils et Procedes d’Identification, Montalieu Vercieu, France) following manufacturers’ instructions. Commercial agglutinating antiserum (Denka Seiken, Tokyo, Japan) was used to serotype Salmonella isolates (Krieg NR, Holt JG (1984) Bergey’s Manual of Systematic Bacteriology. Baltimore: Williams and Wilkins. pp 427–58)