Genetic diversity of Trichomonads from Milu deer (Elaphurus davidianus) in China

The Milu deer, or Père David's deer (Elaphurus davidianus), a rare endemic species in China, represents a case of successful reintroduction of a species previously considered extinct in the wild. Trichomonads, protozoan symbionts capable of infecting vertebrates, are transmitted via the fecal-oral route; they are a subgroup of Parabasalia and include some pathogenic species that pose zoonotic risks. Until now, data on the diversity and prevalence of trichomonads in Chinese Milu deer have not been reported. To better understand the colonization status of trichomonads, fecal samples from 112 Milu deer across five nature reserves in China were collected. The ITS-1/5.8S/ITS-2 sequences were amplified using PCR to investigate the colonization rate of trichomonads and to assess evolutionary relationships and genetic characteristics through phylogenetic analysis. An occurrence of 38.39% was recorded in Milu deer, with sample collection sites (OR = 55.159, 95% CI = 3.166-961.113, p = 0.006), high relative humidity and average annual rainfall (OR = 11.675, 95% CI = 1.747-77.781, p = 0.011) identified as significant risk factors for trichomonads colonization. Undescribed trichomonads from four genera were identified, including Simplicimonas spp., Hypotrichomonas spp., Hexamastix spp., and Tetratrichomonas spp. To our knowledge, this is the first study to report on trichomonads in Milu deer in China. This study aims to enhance understanding of trichomonad colonization and associated risk factors, providing scientific guidance for the ex-situ conservation of Milu deer.