Stomacher lab blender

Collected tissues (lung and spleen) were homogenized with a stomacher lab blender (Seward, England) into single‐cell suspensions in cold SB. Red blood cells (RBCs) were removed from samples by suspension in appropriate amount of RBC lysis buffer (Sigma‐Aldrich, USA) for 3‐5 minutes and washed with SB. Samples were then filtered with 100‐μm cell strainers (Fisher Scientific, China). To isolate mononuclear cells from the lungs, we used Ficoll gradient Histopaque‐1077 (Sigma‐Aldrich, USA) as previously described.16 Isolated cells from the lungs and spleens were counted with trypan blue stain (Sigma‐Aldrich, USA) using a Neubauer haemato‐cytometer chamber (VWR Scientific, USA). For T cell subset determination, isolated cells were stained with the following antibodies: CD3‐FITC or APC, CD4‐PE/CY7 or APC, and CD8‐PE (Biolegend, USA). Stained samples were analysed with a flow‐cytometer FC500 (Beckman Coulter Inc, USA).

Microbial analysis was determined according to the method described in Siripatrawan and Noipha [38 (link)]. Minced pork (25 g) was homogenized in 225 mL of 1 g/100 g peptone and 0.5% (w/v) NaCl for 2 min using a Stomacher Lab Blender (Seward, Model 400 CTF, Bangkok, Thaliand). The prepared samples were spread-plated on Plate Count Agar and incubated at 37 °C for 24 h for total plate counts. For yeasts and molds, the prepared samples were pour-plated on Potato Dextrose Agar and incubated at 25 °C for 5 days. For lactic acid bacteria, the prepared samples were pour-plated on Lactobacillus MRS Agar and incubated at 37 °C for 72 h.