Darine Slama, Habib Mezhoud, Hamouda Babba and Emna Chaker
Background: Many entomological investigations of Culicoides have used the adult specimens as samples for their studies. This is because immature stages are poorly known in most Culicoides species. The lack of correct identification systems of Culicoides larvae has been one of the major obstacles to understanding the field ecology of Culicoides species.
Materials and methods: Culicoides larvae stage was extracted from mud. Larvae stage 2 and 3 were placed in Petri dishes to continue their development to stage IV after pupation.
Larvae stage IV showing the same morphology was divided into 2 groups. First group was grown up to reach the adult stage. Second group of larvae stage IV was used for molecular identification. Thus, DNA was extracted and used as a template for a genus-specific PCR. PCR products were then sequenced and analyzed.
Results: This seems to be the first report identifying Culicoides larvae stage IV by molecular and morphological techniques. By using this method, the sequence of Internal Transcribed Spacer 1 of two Culicoides larvae: C. circumscriptus and C. Puncticollis are identified for the first time.