All samples collected during the experiments were tested for the presence of applied microorganisms, and analyses were carried out under a laminar flow hood using standard microbiological methods. To evaluate the presence and viability of the different microorganisms in the samples (viable and culturable), 100 µL of suspension from each sample was inoculated onto selective media for each microorganism: XLT-4 Agar (Xylose Lactose Tergitol 4, Scharlau, Barcelona, Spain) for S. Typhimurium detection; Baird Parker Agar (Scharlau, Barcelona, Spain) for S. aureus detection; Cetrimide Agar (Scharlau, Barcelona, Spain) for E. aerogenes detection; and Sabouraud Chloramphenicol Agar (Scharlau, Barcelona, Spain) for A. brasiliensis detection. Culture plates were incubated for 24–48 hours at 37 °C. After incubation, suspected bacteria colonies were streaked into a nutrient medium (Scharlab, S.L., Barcelona, Spain) and incubated at 37 °C for 24 hours. Then, API-test (Biomerieux, S.L., Barcelona, Spain) was performed to confirm the bacteria obtained. Filamentous fungi were identified on the basis of macroscopic and microscopic morphologic features (Figure 2 ).
Cetrimide agar
Manufactured by Scharlab
Sourced in Spain, United Kingdom
Cetrimide Agar is a selective and differential culture medium used for the isolation and identification of Pseudomonas aeruginosa from clinical and non-clinical samples. It contains cetrimide, a quaternary ammonium compound, which inhibits the growth of most bacteria, while allowing the growth of Pseudomonas aeruginosa. The medium also contains nutrients and growth factors necessary for the cultivation of Pseudomonas aeruginosa.
Automatically generated - may contain errors
Lab products found in correlation
Nutrient medium, by Scharlab (1 mentions)
Baird parker agar, by Scharlab (1 mentions)
Sabouraud chloramphenicol agar, by Scharlab (1 mentions)
Vitek 2, by bioMérieux (1 mentions)
Api 20ne, by bioMérieux (1 mentions)
Macconkey s agar, by Thermo Fisher Scientific (1 mentions)
Cetrimide agar, by Merck Group (1 mentions)
Pbs solution, by Merck Group (1 mentions)
Model 400, by Seward Medical (1 mentions)
4 protocols using cetrimide agar
1
Microbial Detection Protocols in Samples
2
Detecting Pathogenic Bacteria in Water
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To evaluate the presence of the most common pathogenic bacteria related to FLA, four selective agar media were used: E. coli‐Coliforms Chromogenic Medium (BOE) Conda®; TCBS agar (ISO) VWR Chemicals Prolabo®; SS Agar Merck® and Cetrimide Agar (Pseudomonas selective Agar) Scharlau®. One hundred milliliters of water samples were filtered through 0.45 μm nitrocellulose filters (Pall, Madrid, Spain). Individual membranes were placed into a specific selective medium and incubated for 24h at 37°C depending on the microorganism to be detected.
3
Microbiology Isolation of Adult Clinical Isolates
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We recovered 100 clinical isolates as a microbiology laboratory procedure from the microbiology laboratory at King Abdulaziz Specialist Hospital from February 2021 to February 2022, and all isolates were obtained from adult male/female patients (above 18 years old). Children and pregnant women were not included in this study. All strains were initially recovered on MacConkey’s agar (Oxoid, Basingstoke, UK) and then purified on cetrimide agar (Scharlau, Barcelona, Spain). All isolates were primarily identified by the Vitek 2® system (BioMérieux, Craponne, France) and API 20NE® (BioMérieux, Craponne, France). The genus level was also confirmed via the amplification of the algD gene using the primer pairs (Macrogen, Geumcheon-gu, Seoul, Republic of Korea) listed in Table 1 . Long-term storage of isolates at −84 °C in glycerol brain/heart infusion was undertaken for further tests. This study was performed with ethical approval No. 42-0107 following the regulations of the ethical committee at Taif University.
4
Bacterial Quantification in Food Samples
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Samples were analyzed for mesophilic and psychrophilic bacteria using plate count agar (PCA, Liofilchem) and Pseudomonas spp using cetrimide agar (Scharlau). Each sample (10 g) was diluted 1:10 in sterile distilled water and the mixture was homogenized for 2 min in a stomacher (model 400, Seward, UK). After serial dilutions in PBS solution (Merck), aliquots of 1 ml were inoculated in PCA and cetrimide agar, and the plates were incubated at 37 ± 2 °C for 48 h, 7 ± 2 °C for 7-10 days and 44 ± 2 °C for 48 h to determine mesophilic bacteria, psychrophilic bacteria and Pseudomonas spp., respectively. All analyses were run in duplicate.
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