Safety and Efficacy of a Recombinant Newcastle Disease Virus Expressing Cecropin AD for Diarrhea Reduction and Intestinal Health Improvement in Domestic Pigeons

Diarrhea and impaired intestinal development are major constraints in newly hatched domestic pigeons raised for meat production, leading to reduced growth performance and increased reliance on antimicrobial interventions. Cecropin AD (CAD) is a broad-spectrum antimicrobial peptide with potential activity against bacterial pathogens and may represent an alternative strategy for improving intestinal health in poultry. In this study, we evaluated the safety and biological effects of a recombinant Newcastle disease virus (NDV) expressing CAD (rTS-CAD3) in one-day-old domestic pigeons. Pigeons were inoculated with rTS-CAD3, parental TS09-C, or PBS and monitored for growth performance, diarrhea incidence, viral RNA persistence, inflammatory responses, intestinal bacterial abundance, gut microbiota composition, intestinal morphology, and epithelial barrier function. rTS-CAD3 caused no mortality, detected only transient viral RNA, and did not induce excessive inflammatory responses. Compared with TS09-C and PBS controls, rTS-CAD3 significantly increased body weight gain and average daily gain while reducing diarrhea incidence. Importantly, rTS-CAD3 decreased intestinal Escherichia coli abundance, reshaped the gut microbiota by reducing Proteobacteria and Escherichia–Shigella while increasing Firmicutes and Lactobacillus, improved villus height and the villus-to-crypt ratio, and enhanced occludin expression. These findings demonstrate that NDV-vectored CAD delivery can reduce intestinal bacterial burden, increase occludin expression, and alleviate diarrhea in domestic pigeons. The rTS-CAD3 represents a promising antimicrobial peptide-based strategy for reducing spontaneous diarrhea and dependence on conventional antibiotics in poultry production.