Abstract:: Objective To investigate the characteristics of gut microbiota dysbiosis in hypothermia among infants with very low low birth weight (VLBW). Methods In this prospective cohort study, preterm infants with gestational age<32 weeks and weight<1.5 kg were enrolled. Additionally, fecal samples were collected within 0 days after admission to the intensive care unit. Based on rectal temperatures upon admission, the VLBW infants were divided into the hypothermia group (body temperature<36 ℃) and the normothermia group (body temperature≥36.5 ℃). 16S rRNA gene sequencing was utilized to evaluate the gut microbial diversity, composition and biomarker. Results A total of 19 patients with hypothermia were enrolled from April 2022 to December 2022, alongside an additional cohort of 15 normothermic patients who were admitted during the same study period. Compared to the normothermia group, the hypothermia group exhibited a significant increase in the abundance of p_Bacteroidota, c_Clostridia and c_Bacteroidia in the hypothermia group (P=0.031, 0.042, 0.031, respectively). Furthermore, the relative abundances of Faecalibaculum, Dubosiella, Blautia and Faecalibaculum_rodentium were significantly higher in the hypothermia group (P=0.008, 0.008, 0.005, and 0.008, respectively). The overall microbial diversity exhibited a notable disparity between the hypothermia and normothermia groups. CCA results showed that the r2 value of cesarean section was the largest (r2 =0.4484, P=0.001), and cesarean section had the greatest impact on the structure of gut microbiota. Conclusion VLBW in the hypothermia group has imbalance in the diversity and composition of gut microbiota, which suggest that the dominant genus Faecalibaculum is a potential a biomarker for hypothermia group.