Triple sugar iron (TSI), Simmons citrate, lysine iron agar (LIA) and urea agar (Oxoid) slants were prepared according to the manufacturer’s instructions and inoculated with a 24 h nutrient agar-grown culture of the isolates incubated at 37 °C. The surface of the agar slant was inoculated using a sterile inoculating loop while a stab was made at the center of the slant using a sterile inoculating needle. The tubes were then incubated under aerobic conditions at 37 °C for 24 to 48 h. Tubes exhibiting alkaline slant and acidic butt with H2S production on TSI slants, purple color in butt of LIA tube, blue color development on slant of citrate agar and no color change on urea indicated positive results for Salmonella spp. Isolates showing biochemical reaction consistent with Salmonella spp. were further confirmed using molecular based method.
Urea agar
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Urea agar is a microbiological culture medium used for the detection and identification of microorganisms capable of hydrolyzing urea. It is a solid culture medium that contains urea as the primary carbon and nitrogen source. The hydrolysis of urea by certain microorganisms results in the production of ammonia, which increases the pH of the medium and can be detected by the color change of the pH indicator included in the formulation.
Automatically generated - may contain errors
Lab products found in correlation
Simmon citrate agar, by Thermo Fisher Scientific (2 mentions)
Xylose lysine deosxycholate agar, by Thermo Fisher Scientific (1 mentions)
Lysine agar, by Thermo Fisher Scientific (1 mentions)
Selenite f enrichment broth, by Thermo Fisher Scientific (1 mentions)
Kligler iron agar, by Thermo Fisher Scientific (1 mentions)
Macconkey agar, by Thermo Fisher Scientific (1 mentions)
Triple sugar iron, by Thermo Fisher Scientific (1 mentions)
Buffer peptone water, by Thermo Fisher Scientific (1 mentions)
Sheep blood agar, by Thermo Fisher Scientific (1 mentions)
Triple sugar iron agar, by Thermo Fisher Scientific (1 mentions)
Selenite f broth, by Thermo Fisher Scientific (1 mentions)
Api 10s, by bioMérieux (1 mentions)
Xylose lysine deoxycholate agar, by Thermo Fisher Scientific (1 mentions)
Macconkey agar plate, by Thermo Fisher Scientific (1 mentions)
5 protocols using urea agar
1
Biochemical Identification of Salmonella
2
Isolation and Identification of Shigella
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MacConkey agar, Xylose Lysine Deosxycholate agar and Selenite F enrichment broth (Oxoid, England) were used for isolation of Shigella. Culture negative specimens on primary solid media were sub-cultured from the enrichment broth to primary solid media to improve recovery of the isolates. All inoculated media were incubated at 37 °C for 18–24 h. After overnight incubation, non-lactose fermenters were further identified by biochemical tests using appropriate media namely: Kligler Iron Agar for carbohydrate fermentation test, Urea agar for the urea utilization test, tryptophan broth for Indole test, Simmon Citrate agar for citrate utilization, Motility agar for motility test, Lysine agar for lysine utilization test (all Oxoid, England).
3
Isolation and Identification of Bacteria from Brain Tissue
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Bacteria were isolated and identified, as previously described [25 ]. Briefly, pre-enrichment of brain tissue used Buffer Peptone Water (Oxoid®, UK) incubated at 37°C for 16-18 h. One-tenth mL of pre-enrichment medium was transferred to Modified Semisolid Rappaport-Vassiliadis medium (LabM®, UK) and incubated at 41.5°C for 24 h. Furthermore, 1 mL was transferred to MKTTn broth (LabM®, UK) and incubated aerobically for 24 h. The samples were then streaked onto XLD (LabM®, UK) and SS (Oxoid®, UK) agar plates and incubated at 37°C for 24 h, aerobically. Typical colonies were identified by biochemical tests (Urea agar, Triple sugar iron, and Lysin iron) (Oxoid®, UK).
4
Phenotypic Confirmation of E. coli
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About 10% of the isolated colonies were purified and confirmed as E. coli using phenotypic tests including citrate production (Simmon Citrate Agar–CM 0155, Oxoid, UK), acid and gas production (Triple Sugar Iron slant agar–CM 0277, Oxoid, UK), indole production, motility (Sulphur Indole Motility media–CM 0435, Oxoid, UK) and urea production (Urea Agar–CM 0053, Oxoid, UK). The sterile distilled water was plated and used as negative controls. Duplicate plates of the hand-rinse samples were inoculated and colonies enumerated. Where the E. coli colonies were too numerous to count, a 10−1 serial dilution was inoculated the next day to grow and enumerate them. The mean counts of E. coli for each duplicate plate were used in the analysis. Stakeholder interviews were transcribed verbatim and transcripts were checked by the author for completeness and accuracy. The questionnaire was validated through a pre-test in Dodowa, a similar study setting as Ningo-Prampram.
5
Salmonella Isolation from Stool Samples
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A matchstick head-size sample of stool was inoculated in selenite F broth (Oxoid, UK, catalog number: 2300631) and aerobically incubated overnight at 37 °C for 18-24 hours. The top layer (1 ml) of an overnight culture was spun at 20,000 g for 5 minutes and the pellet was sub-cultured on Xylose Lysine Deoxycholate (XLD) agar (Oxoid, UK, catalog number: 2547703). An aliquot of the selenite broth was also frozen for molecular detection (below). Presumptive Salmonella colonies were cultured onto sheep blood agar (Oxoid, UK, catalog number: 2910831) and MacConkey agar plates (Oxoid, UK, catalog number: 2529552) and incubated aerobically at 37°C for 18-24 hours. Salmonella colonies were then distinguished from other enteric bacteria (i.e. Citrobacter and Serratia) using triple sugar iron agar (Oxoid, UK, catalog number: 1882283) and Urea agar (Oxoid, UK, catalog number: 1779617) biochemical tests. Further Salmonella identification was determined using API® 10S (bioMérieux, France, catalog number: 1007181060) according to the manufacturer's instructions.
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