Comparison of Two Methods for the Isolation of Genomic DNA from Cyathostomin Adult Parasites

Juliana Bana Ishii1*, Amilcar Arenal, Yanise Chicoy, Marcelo Beltrão Molento


1Laboratory of Parasitic Diseases, Federal University of Parana. Curitiba, PR, Brazil

2Laboratory of Biochemistry, University of Camaguey. Camaguey, Cuba

3National Institute of Science and Technology, INCT- Pecuaria. Belo Horizonte, MG. Brazil



*Sorumlu Yazar / Corresponding Author:

Juliana Bana Ishii

e-mail: julianabana.ishii@gmail.com

Geliş Tarihi / Received:

12 October 2016

Kabul Tarihi / Accepted:

07 December 2016

Key Words:

DNA extraction, genomic material, horse, Strongylida;

Abstract

Cyathostomins are the most common and important group of large intestine nematodes, infecting horses worldwide. The current control strategy is associated with the development of anthelmintic resistance, which has been reported worldwide. Therefore, experiments with this family of parasites have become progressively important to provide their monitoring and control strategies. The aim of the present study was to propose a faster and more economic assay for isolation of genomic DNA from the adult stage of Cyathostomin parasites than reported. Adult parasites were collected from a single horse from a farm in São José dos Pinhais, PR, Brazil, and were identified. Genomic DNA was isolated from ten individual female adult parasites using a standardized procedure developed. Then, extraction from ten individual female was carried out by another DNA extraction method. DNA concentration from both methods were measured and compared. We obtained a good DNA quality with this standardized procedure. As a result of this analysis, we propose a modified phenol-chloroform method, which will contribute to assays that require DNA extraction from adult worms for genomic DNA sequences of cyathostomin, or species-specific identification.