Improved and simplified method for aseptic isolation of nematodes and nematode-endosymbiotic bacteria from pine seedlings

Pine wilt disease (PWD), caused by the pinewood nematode (PWN), Bursaphelenchus xylophilus, significantly impacts pine species and poses a broader ecological concern. An understanding of these nematode-associated microbes is essential for formulating sustainable PWD management strategies. We introduce a streamlined method for the aseptic extraction of B. xylophilus • A simplified, aseptic method for extracting B. xylophilus from pine seedlings, offering a modern alternative to traditional Baermann funnel method. • Utilization of a specialized sterile syringe assembly setup, ensuring controlled and optimal nematode isolation. • Method validation achieved through the successful isolation and identification of bacterial strains as nematode endosymbionts.