Bacterial burden were also assessed.<h4>Results</h4>We identified pervasive NETosis as a pathological hallmark of non-healing anal fistula wounds, accompanied by a surge in pro-inflammatory cytokines (IL-2, IL-5, IL-6, IL-12, TNF-α) and a ~15.5-fold increase in bacterial load compared to controls. Here, IL5 is linked to Anal fistula.