First Molecular Evidence of Alaria Alata Different Development Stages in Latvia

The genus Alaria includes seven species but only Alaria alata is widespread throughout all the Europe. In present research 43 adult samples, 20 mesocercaria samples and ten egg samples were analyzed utilizing molecular biology methods. QIAamp DNA Mini Kit (Protocol: DNA purification from tissues, Qiagen, Germany) and DNeasy Blood & Tissue Kit (Protocol: Purification of total DNA from animal tissues, Qiagen, Germany) spin-column protocols were approbated and modified to find out the suitable one for all development stages of A. alata. Extracted DNA was amplified and sequenced using primers used in previous studies. Two new primer pairs were generated based on obtained sequences and data from GeneBank database. Scientific impact: 1. This is the first try to extract A. alata DNA from their eggs. 2. Dneasy Blood & Tissue Kit procedure with increased amount of proteinase K (25μl) and increased incubation time (15h) were most suitable for DNA purification from different A. alata development stages. 3. With newly designed primers 28.1% more sequences were obtained. 4. This study demonstrate the necessity to improve the efficacy of the DNA extraction technique prior further studies