Liver, spleen, and mesenteric lymph node (MLN) samples were homogenized in PBS before being placed on blood agar (Oxoid, UK) for aerobic incubation, or on anaerobic agar (Oxoid, UK) for anaerobic incubation immediately after collection. The samples were then incubated in corresponding anaerobic chambers and aerobic atmospheres for 48 h at 37°C. The sterile swab technique was used to culture the peritoneal surface. If bacteria were present, BT was indicated. The number of living bacteria was calculated and expressed in colony forming units per gram of organ tissue (cfu/g).
Anaerobic agar
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, Slovakia
Anaerobic agar is a solid medium used for the cultivation and isolation of anaerobic bacteria. It provides the necessary nutrients and conditions to support the growth of microorganisms that thrive in the absence of oxygen.
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Lab products found in correlation
2 protocols using anaerobic agar
1
Bacterial Load Quantification in Tissues
2
Microbial Analysis of Sonication Fluids and Urine
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All sonication fluids and urine samples (1–100) were inoculated into a set of solid media: Blood Agar with 7% sheep’s blood (Oxoid, UK), Endo Agar (Imuna Pharm, Šarišské Michaľany, Slovakia), Wilkins Chalgren Anaerobic Agar with 7% sheep’s blood and vitamin K (WCHA, Oxoid, Hampshire, UK). UCs, DJCs and urine samples were inoculated in addition into Blood Agar with 10% of NaCl, Blood Agar with Amikacine (32 mg/L) (Oxoid, Hampshire, UK), and UriSelect 4 (Bio-Rad, Hercules, CA, USA). Plates were assessed for microbial growth after cultivating at 37 °C for 48 h. The WCHA was cultivated in an anaerobic atmosphere (80% N2, 10% CO2, and 10% H2; Anaerobic Work Station Concept 400, Ruskinn Technology, Bridgend, UK) at 37 °C for 7 days. All isolated strains were identified biochemically or using MALDI-TOF MS (Biotyper with FlexControl 3.4 software, Bruker Daltonics, Billerica, MA, USA), according to the manufacturer’s instructions. For details see [30 (link)].
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