First report of amblyomma sp. Collected from varanus salvator in baluran national park identified by DNA barcoding

Ticks are one of crucial vectors of disease and parasites of animals. Traditionally, identification of tick species has been conducted using morphological features. This identification method has some limitation such as limited by the condition of samples, little variation among specimens, and difficult to correctly identify the specimens subadult stages. A new, fast, and valid identification method is required to overcome this traditionally species identification. DNA barcoding apply a specific fragment of the mitochondrial gene, cytochrome oxidase c subunit I (COI), to identify species and has been successfully used in many species and other taxa level. This method aids both taxonomists and non-taxonomists to recognize the species of Ixodida based on DNA sequences rather than morphological traits. In this study, we applied DNA barcoding to identify hard tick species which invested on Varanus salvator in Baluran National Park, East Java, Indonesia. By analyzing 648-660 bp of cytochrome c oxidase subunit I gene sequences obtained from hard tick specimens, COI barcoding showed that the hard ticks were identified as Amblyomma sp. All of sequences from specimens were identical. BLAST analysis showed that the closest species with specimens was A. testudinarium with 95% identity. The lowest K2P genetic distance between specimens in this study and A. testudinarium obtained from GenBank was 5.4%. The NJ tree analysis using reference COI sequences from genus Amblyomma and from genus member in Ixodidae also demonstrated that hard tick specimens was Amblyomma sp. However, there are still limitation in the database of COI sequence of some species of hard ticks, and further research is required to resolve this issue.