Determination of the prevalence cestode with assignment of Mitochondrial DNA barcodes

Abstract

Background: Mitochondrial deoxyribonucleic acid sequences are widely used in phylogenetic analysis and taxonomy for species identification. Scientists suggest using the cytochrome c-oxidase subunit 1 (cox1) gene in the mitochondrial genome as a barcode. The use of the mitochondrial genome gene cox1 as a DNA barcode is considered optimal because it contains universal primers and more phylogenetic information than other genes. Aim: Therefore, this study was performed to determine the presence of tapeworm infection in a dog population using the mitochondrial cox1 gene DNA barcode. Materials and Methods: A total of 1242 feces were collected from 21 provinces of Mongolia by random instant selection. Genomic DNAs from fecal samples were prepared using QIAamp DNA Stool Mini kit (Qiagen, Tokto, Japan). The mitochondrial 12S ribosomal RNA genes (12S rRNA) and cytochrome c oxidase subunit I (cox1) were amplified using primers as described. Template DNA samples were prepared using DyeTerminator Cycle Sequencing kit and all amplicons were sequenced in both directions on a 3100-Advant Genetic Analyzer. Results: Taenia multiceps infection was detected in 5.2% (64/1242) of the cox1 and 12S rRNA gene nucleotide sequences from total samples analyzed with the sequence stored in Genbank. The prevalence of Taenia multiceps infection in the natural zone was found in the taiga (12.5%), forest-steppe (6.6%), steppe (4.7%), desert steppe (3.4%), and desert (2.3%). The highest frequency of Taenia multiceps infestation was 13.2% (7/53) in Khuvsgul province. The genetic distance between Taenia multiceps and T. hydatigena species is relatively large, 1.140-1.168; the haplotype AB792722 of Taenia multiceps is genetic distance with haplotypes AB792721 and AB792724 is relatively close, 0.002. Conclusion: Our study showed that there is a high prevalence of infection with cyclophyllid cestodes in sheepdogs throughout Mongolia. Therefore, implementation of a comprehensive surveillance program, including evaluation of sheepdogs and their owners and wild animals in all ecological zones should be considered.