All pre-enriched samples were incubated for 24 h at 37°C. After that, 0.1 ml of the incubated broth was added to 10 ml Rappaport Vassilidis and incubated at 42°C for 24 h. Then, one loopful was streaked onto Salmonella Shigella agar (SS agar: Oxoid®, CM 0099, Ltd, UK) and incubated at 37°C for 24 h for Salmonella isolation. For isolation of other Gram-negative bacteria of family Enterobacteriaceae and Pseudomonas spp., one loopful from BPW-enriched broth inoculated into MacConkey agar (Oxoid®, CM 0007, Ltd, UK) plates and incubated at 37°C for 24 h. The pure separate colonies were picked up and inoculated into nutrient agar slope and incubate at 37°C for 24 h. The identification of bacterial colonies was based on colonial morphology, pigmentation, and Gram staining and biochemical reaction tests, according to Forbes et al. [22 ].
Cm0099
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The CM0099 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is designed to perform a core function within the laboratory setting. The specific details and intended use of this product are not available in this factual and unbiased response.
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5 protocols using cm0099
1
Isolation and Identification of Salmonella and Enterobacteriaceae
2
Isolation and Identification of Salmonella
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Isolation and identification of Salmonella spp. were performed using procedures previously described by Addo et al. (18 (link)). 1 ml aliquots stock homogenate prepared earlier were transferred into 10 ml Rappaport–Vassiliadis Broth for enrichment. Samples in Rappaport–Vassiliadis Broth (Oxoid, CM0669) were incubated at 37°C for 24 h. 0.1 ml of the enriched samples were then streaked onto Salmonella–Shigella Agar (SSA) (Oxoid, CM0099) and incubated at 37°C for 24 h. Cream colonies with black centers on the SSA presumed to be Salmonella spp. were purified on nutrient agar and confirmed using Gram staining, analytical profile index (API) (20E, Biomérieux, France), and Salmonella latex agglutination.
3
Blood Culture and Bacterial Isolation
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Two milliliters of venous blood sample taken from patients was inoculated into 20 ml brain-heart infusion broth (BHI) (CM1135; Oxoid Ltd., England, UK). The culture bottles were incubated at 37 °C for 7 days and examined daily for evidence of bacterial growth, including turbidity and hemolysis. If bacterial growth was observed, a subculture was made after 24 h on blood agar (CM0271, Oxoid Ltd., England UK), Salmonella Shigella agar (SS) (CM0099, Oxoid Ltd., England, UK), and MacConkey agar (CM0007, Oxoid Ltd., England, UK).
4
Determination of Total Plate Count
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The reported method of Liu et al. [20 (link)] was employed for determination of the total plate count. Briefly, 1 g of powder sample was homogenized at 5000 g using a homogenizer (Ultra-turrax T25 blade-type homogenizer; Ika-Werk, Staufen, Germany) in 9.0 mL sterile 0.1% peptone water for 30 s and then serial dilution was carried out. Aliquots in a quantity of 1 mL were plated, and nutrient agar (CM003; Oxoid Ltd., Basingstoke, Hampshire, UK) and Malt Extract Agar (CM 0099; Oxoid), being suitable media for the growth of total aerobic bacteria and fungi, respectively, were used for media preparation. Plated aliquots for total aerobic bacteria were incubated at 37 °C for 24 h, whereas fungi-plated aliquots were incubated at 25 °C for 5 days. A colony counter (LM-10; Analab, New Delhi, India) was used to enumerate the microbial population, and the counts of the microorganisms were recorded and the readings were taken from 10−3 serial dilution.
5
Bacterial Culture and Serological Testing
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Venous blood samples were taken from patients by venipuncture. This was followed by inoculation of 7 mls or 3 mls of blood into 20 mls of sterile brain-heart infusion broth (BHI) (CM1135; Oxoid Ltd., England, UK) The culture bottles were incubated at 37°C for 7 days and examined daily for evidence of bacterial growth, including turbidity and hemolysis. If bacterial growth was observed, a subculture was made on Xylene Lysine Deoxycholate agar (XLD) (CM0271, Oxoid Ltd., England UK) and Salmonella Shigella agar (SSA) (CM0099, Oxoid Ltd., England, UK). Three(3) mls of blood sample used to run the Salmonella Typhi and Paratyphi IgG/IgM serological test.
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