Abstract
Background:
The Setaria nematode, classified within the order Spirurida and family Setariidae, predilection in the peritoneal cavity of its host, with mosquitoes acting as vectors for transmitting microfilariae to cattle, its primary host.

Aim:
This study investigates the morphology and molecular characteristics of Setaria digitata in Aceh cattle and evaluates the effectiveness of three staining solutions lactophenol, glycerol, and lugol 2% for visualizing adult worms.

Methods:
A total of 124 blood serum and adult worm samples were collected from Aceh cattle at slaughterhouses in Banda Aceh and Bireuen. Microfilariae detection was conducted using the Knott test, while worm morphology was assessed via Polymerase Chain Reaction (PCR) amplification of the 12S rDNA and cox1 genes.

Results:
Each staining solution enhanced worm morphology visualization with varying benefits. The lugol 2% solution offered a complete view of morphology but lacked detail for internal structures. Lactophenol improved visibility of both morphology and internal organs, though sample integrity declined quickly. Glycerol displayed both the morphology and internal structures with clarity, making it the most effective option overall. Molecular amplification results showed a 450 bp band for the 12S rDNA gene and a 680 bp band for the cox1 gene in S. digitata.

Conclusion:
Glycerol proved superior for maintaining morphology and visualizing internal structures of S. digitata. Molecular profiling using the 12S rDNA and cox1 genes effectively confirmed the identity of Setaria digitata, underscoring the value of these genetic markers for species identification.

Key words: Morfology, Molecular, Setaria, Aceh cow, Staining solutions