Abstract: 【Objective】 This article aims to identify the main pathogens of calf diarrhea in a large-scale cattle farm in Xinjiang. 【Methods】 Virus detection was performed on 14 diarrhea calf feces using RT-PCR. Besides, bacteria were isolated from the liver tissues of four dead calves, cultured and identified, and mouse pathogenicity and drug sensitivity tests were conducted on these bacteria. Afterwards, the 16 S rDNA gene fragment of the isolated pathogenic bacteria was amplified and sequenced, and the sequencing results were searched for homologous sequences using BLAST in NCBI for genetic evolution analysis. 【Results】 The pathogens of diarrhea in the cattle farm were Bovine coronavirus (BCoV), Bovine rotavirus (BRV), Escherichia coli (E.coli), and Klebsiella pneumoniae (K.pneumoniae). Among the 14 feces, the detection rate of BCoV was 64.29%, and the detection rate of BRV was 50.00%; Among the four liver tissues, the detection rate of E. coli was 100.00%, and the detection rate of K.pneumoniae was 50.00%. The two isolated pathogenic bacteria E.coli and K. pneumoniae were sensitive to amikacin and resistant to all 16 other antibiotics. 【Conclusion】 The main pathogens causing calf diarrhea in this cattle farm are BCoV, BRV, E.coli, and K.pneumoniae, with mixed infection of viruses and bacteria. Amikacin can be used as the preferred drug for the prevention and treatment of E. coli and K. pneumoniae.