Ek-Huchim Juan Pablo, Aguirre-Macedo Ma Leopoldina, Améndola-Pimenta Monica, Vidal-Martínez Victor Manuel, Pérez-Vega Juan Antonio, Simá-Alvarez Raúl, Jiménez-García Isabel, Zamora-Bustillos Roberto, Rodríguez-Canul Rossanna
Laboratorio de Inmunología y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN) Unidad Mérida, Carretera Antigua a Progreso Km. 6, 97310, Mérida, Yucatán, Mexico.
Laboratorio de Patología Acuática, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN) Unidad Mérida, Carretera Antigua a Progreso Km. 6, 97310, Mérida, Yucatán, Mexico.
Parasit Vectors. 2017 Aug 2;10(1):372. doi: 10.1186/s13071-017-2304-4.
The protozoan Perkinsus marinus (Mackin, Owen & Collier) Levine, 1978 causes perkinsosis in the American oyster Crassostrea virginica Gmelin, 1791. This pathogen is present in cultured C. virginica from the Gulf of Mexico and has been reported recently in Saccostrea palmula (Carpenter, 1857), Crassostrea corteziensis (Hertlein, 1951) and Crassostrea gigas (Thunberg, 1793) from the Mexican Pacific coast. Transportation of fresh oysters for human consumption and repopulation could be implicated in the transmission and dissemination of this parasite across the Mexican Pacific coast. The aim of this study was two-fold. First, we evaluated the P. marinus infection parameters by PCR and RFTM (Ray's fluid thioglycollate medium) in C. virginica from four major lagoons (Términos Lagoon, Campeche; Carmen-Pajonal-Machona Lagoon complex, Tabasco; Mandinga Lagoon, Veracruz; and La Pesca Lagoon, Tamaulipas) from the Gulf of Mexico. Secondly, we used DNA sequence analyses of the ribosomal non-transcribed spacer (rNTS) region of P. marinus to determine the possible translocation of this species from the Gulf of Mexico to the Mexican Pacific coast.
Perkinsus marinus prevalence by PCR was 57.7% (338 out of 586 oysters) and 38.2% (224 out of 586 oysters) by RFTM. The highest prevalence was observed in the Carmen-Pajonal-Machona Lagoon complex in the state of Tabasco (73% by PCR and 58% by RFTM) and the estimated weighted prevalence (WP) was less than 1.0 in the four lagoons. Ten unique rDNA-NTS sequences of P. marinus [termed herein the "P. marinus (Pm) haplotype"] were identified in the Gulf of Mexico sample. They shared 96-100% similarity with 18 rDNA-NTS sequences from the GenBank database which were derived from 16 Mexican Pacific coast infections and two sequences from the USA. The phylogenetic tree and the haplotype network showed that the P. marinus rDNA-NTS sequences from Mexico were distant from the rDNA-NTS sequences of P. marinus reported from the USA. The ten rDNA-NTS sequences described herein were restricted to specific locations displaying different geographical connections within the Gulf of Mexico; the Carmen-Pajonal-Machona Pm1 haplotype from the state of Tabasco shared a cluster with P. marinus isolates reported from the Mexican Pacific coast.
The rDNA-NTS sequences of P. marinus from the state of Tabasco shared high similarity with the reference rDNA-NTS sequences from the Mexican Pacific coast. The high similarity suggests a transfer of oysters infected with P. marinus from the Mexican part of the Gulf of Mexico into the Mexican Pacific coast.
原生动物马氏帕金虫(Perkinsus marinus)(Mackin、Owen和Collier,1978年)可引发美国牡蛎(Crassostrea virginica Gmelin,1791年)的帕金虫病。这种病原体存在于墨西哥湾养殖的弗吉尼亚牡蛎中,最近在墨西哥太平洋沿岸的掌状牡蛎(Saccostrea palmula)(Carpenter,1857年)、科尔特斯牡蛎(Crassostrea corteziensis)(Hertlein,1951年)和太平洋牡蛎(Crassostrea gigas)(Thunberg,1793年)中也有报道。供人类食用的新鲜牡蛎的运输和再养殖可能与这种寄生虫在墨西哥太平洋沿岸的传播和扩散有关。本研究有两个目的。第一,我们通过聚合酶链式反应(PCR)和雷氏流体巯基乙酸盐培养基(RFTM)评估了来自墨西哥湾四个主要泻湖(坎佩切州的特米诺斯泻湖、塔巴斯科州的卡门 - 帕霍纳尔 - 马乔纳泻湖群、韦拉克鲁斯州的曼丁加泻湖和塔毛利帕斯州的拉佩斯卡拉泻湖)的弗吉尼亚牡蛎中马氏帕金虫的感染参数。第二,我们对马氏帕金虫核糖体非转录间隔区(rNTS)进行DNA序列分析,以确定该物种从墨西哥湾到墨西哥太平洋沿岸可能的迁移情况。
通过PCR检测,马氏帕金虫的患病率为57.7%(586只牡蛎中有338只),通过RFTM检测为38.2%(586只牡蛎中有224只)。在塔巴斯科州的卡门 - 帕霍纳尔 - 马乔纳泻湖群中观察到最高患病率(PCR检测为73%,RFTM检测为58%),四个泻湖的估计加权患病率(WP)小于1.0。在墨西哥湾样本中鉴定出10个独特的马氏帕金虫核糖体DNA - NTS序列(本文称为“马氏帕金虫(Pm)单倍型”)。它们与来自GenBank数据库的18个核糖体DNA - NTS序列具有96 - 100%的相似性,这些序列来自16个墨西哥太平洋沿岸感染样本和两个来自美国的样本。系统发育树和单倍型网络显示,来自墨西哥的马氏帕金虫核糖体DNA - NTS序列与美国报道的马氏帕金虫核糖体DNA - NTS序列距离较远。本文描述的10个核糖体DNA - NTS序列局限于墨西哥湾内显示不同地理联系的特定位置;来自塔巴斯科州的卡门 - 帕霍纳尔 - 马乔纳Pm1单倍型与墨西哥太平洋沿岸报道的马氏帕金虫分离株共享一个聚类。
来自塔巴斯科州的马氏帕金虫核糖体DNA - NTS序列与来自墨西哥太平洋沿岸的参考核糖体DNA - NTS序列具有高度相似性。这种高度相似性表明感染马氏帕金虫的牡蛎从墨西哥湾的墨西哥部分转移到了墨西哥太平洋沿岸。