show Abstracthide AbstractTo assess the effectiveness of high-throughput sequencing technology in detecting both feed raw materials and potential eukaryotic pathogens, this study evaluated the eukaryotic composition of raw materials using three primer pairs of eukaryotic primers. Our results showed significant variations in species annotation among the sequences obtained from different primers. Specifi-cally, the 18S rRNA gene primer pairs (Uni18S and Uni18SR, and TAReuk454FWD1 and TAReukREV3) were able to detect a wide range of species and particularly effective for identifying fungi, whereas the 12S rRNA gene primer pair (AcMDB07-F and AcMDB07-R) provided more detailed identifications of multicellular ingredient sources.