Uribe Nelson, Becerra Wlda Margarita, Velásquez Luz Elena
Línea de Investigación en Epidemiología, Diagnóstico y Control de Enfermedades causadas por Trematodos, GIEM, Facultad de Salud, Universidad Industrial de Santander.
Semillero de Investigación en Enfermedades Parasitarias y Parasitología (SIEPA), Facultad de Ciencias Básicas, Universidad de Pamplona, Pamplona, Colombia.
Biomedica. 2014 Oct-Dec;34(4):598-604. doi: 10.1590/S0120-41572014000400013.
Fasciolosis is the disease transmitted by vectors with the highest latitudinal, longitudinal, and altitudinal distribution due to the colonizing capacity of the parasite Fasciola hepatica and its intermediate hosts, Lymnaeidae mollusks. These snails are under research due to their epidemiological importance, but their taxonomic identification is difficult given their interspecific phenotypical similarity. For this reason, there is uncertainty regarding Lymnaea cousini -a host of F. hepatica in Colombia- due to the morphological similarity it has with Lymnaea meridensis , recently described for Venezuela.
To confirm with the COI marker (ADNmt) the taxonomic status of individuals morphologically identified as L. cousini from Nariño, Norte de Santander, and Santander (Colombia), deposited in the Vector Mollusks Collection VHET No. 37 of Universidad de Antioquia.
The amplification of the mitochondrial COI required total DNA extraction of each individual´s foot using the DNeasy Blood and Tissue Kit (Qiagen®). Products amplified were sent for sequencing to Macrogen Inc., Korea. Twenty seven sequences generated in this research were compared to sequences published in the GenBank, including sequences of the type locality of L. cousini .
Two new haplotypes of L. cousini were obtained for Colombia. Specimens from Nariño correspond to haplotype A, referenced for Ecuador, and specimens from Santander and Norte de Santander belong to a new haplotype we called haplotype D.
By using the mitochondrial COI marker, we confirmed that the species under study did correspond to L. cousini . The number of known haplotypes of the species for Colombia has been duplicated and its geographical distribution has been extended to the southwest and northeast of the Colombian high Andean region.
由于肝片吸虫及其中间宿主椎实螺科软体动物的定殖能力，肝片吸虫病是由媒介传播的疾病中具有最高纬度、经度和海拔分布范围的疾病。这些蜗牛因其流行病学重要性而受到研究，但鉴于它们种间表型相似，其分类鉴定较为困难。因此，由于哥伦比亚的肝片吸虫宿主库西椎实螺与委内瑞拉最近描述的梅里登斯椎实螺形态相似，其分类存在不确定性。
利用细胞色素氧化酶亚基I（COI）标记（ADNmt），确认安蒂奥基亚大学媒介软体动物收藏VHET No. 37中保存的、来自纳里尼奥、北桑坦德和桑坦德（哥伦比亚）的形态学上鉴定为库西椎实螺的个体的分类地位。
使用DNeasy Blood and Tissue Kit（Qiagen®）从每个个体的足部提取总DNA，用于线粒体COI的扩增。扩增产物送往韩国Macrogen公司进行测序。将本研究中产生的27个序列与GenBank中公布的序列进行比较，包括库西椎实螺模式产地的序列。
获得了哥伦比亚库西椎实螺的两个新单倍型。来自纳里尼奥的标本对应于厄瓜多尔的A单倍型，来自桑坦德和北桑坦德的标本属于一个新的单倍型，我们称之为D单倍型。
通过使用线粒体COI标记，我们确认所研究的物种确实是库西椎实螺。该物种在哥伦比亚已知单倍型的数量增加了一倍，其地理分布扩展到了哥伦比亚安第斯山脉高地的西南部和东北部。
