Galon Eloiza May, Macalanda Adrian Miki, Sugi Tatsuki, Hayashida Kyoko, Kawai Naoko, Kidaka Taishi, Ybañez Rochelle Haidee, Adjou Moumouni Paul Franck, Ringo Aaron Edmond, Li Hang, Ji Shengwei, Yamagishi Junya, Ybañez Adrian, Xuan Xuenan
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, Japan.
College of Veterinary Medicine and Biomedical Sciences, Cavite State University, Indang 4122, Philippines.
Microorganisms. 2023 Oct 18;11(10):2584. doi: 10.3390/microorganisms11102584.
Molecular assays and capillary electrophoresis sequencing have been used to identify parasites in livestock. The low sample capacity, which increases labor and processing time, is one drawback. Targeted amplicon sequencing (Ampliseq) uses the fast and large sample capacity platform to identify parasites in the target host, overcoming this limitation. DNA was extracted from 162 whole blood samples collected from cattle in three provinces in the Philippines. Using Illumina's Miseq platform, the V4 hypervariable region of the piroplasma 18S rRNA gene was amplified and sequenced. The AMPtk pipeline was used to obtain distinct amplicon sequence variants (ASVs) and the NCBI BLAST non-redundant database was used to assign taxonomy. In total, 95 (58.64%) samples were positive for piroplasma. Using the AMPTk pipeline, 2179 ASVs were obtained. A total of 79 distinct ASVs were obtained after clustering and filtering, which belonged to genera (n = 58), (n = 17), (n = 2), and (n = 2). The ASV top hits were composed of 10 species: , , , sp., , , , , , and sp. Thung Song. The results generated in this study demonstrated the applicability of Ampliseq in detecting piroplasmid parasites infecting cattle in the Philippines.
分子检测和毛细管电泳测序已被用于鉴定家畜中的寄生虫。样本量少会增加人工和处理时间,这是一个缺点。靶向扩增子测序(Ampliseq)使用快速且样本量较大的平台来鉴定目标宿主中的寄生虫,克服了这一局限性。从菲律宾三个省份采集的162份牛全血样本中提取DNA。使用Illumina的Miseq平台,对梨形虫18S rRNA基因的V4高变区进行扩增和测序。使用AMPTk流程获得不同的扩增子序列变体(ASV),并使用NCBI BLAST非冗余数据库进行分类。共有95份(58.64%)样本的梨形虫检测呈阳性。使用AMPTk流程,获得了2179个ASV。经过聚类和筛选后,共获得79个不同的ASV,它们分别属于属(n = 58)、属(n = 17)、属(n = 2)和属(n = 2)。ASV的前几个匹配结果由10个物种组成:、、、种、、、、、和通宋种。本研究产生的结果证明了Ampliseq在检测菲律宾感染牛的梨形虫寄生虫方面的适用性。