Mariné Oliveira Geisi Ferreira, do Couto Melissa Carvalho Machado, de Freitas Lima Marcelo, do Bomfim Teresa Cristina Bergamo
Educacional Foundation Rosemar Pimentel, University Center Geraldo Di Biase - Barra do Piraí, Brazil.
Federal Rural University of Rio de Janeiro, Institute of Biology, Department of Physiological Sciences, Brazil.
Int J Parasitol Parasites Wildl. 2016 Jan 18;5(1):28-33. doi: 10.1016/j.ijppaw.2016.01.004. eCollection 2016 Apr.
Sources of contamination such as animal feces runoff, organic fertilizer application, and the release of partially treated or untreated sewage can lead to the contamination of aquatic environments by Cryptosporidium spp. The quality of mussels as food is closely related to the sanitary conditions of the marine environment where these bivalves are found. Marine mollusks are filter feeders that are able to retain Cryptosporidium oocysts in their tissue, thus functioning as bioindicators. A total of 72 pooled mussel samples of the species Perna perna were collected at two sites (A and B) in the municipality of Mangaratiba, Rio de Janeiro State, Brazil. Sampling involved removal of 30 mussels, from each collection site every month for one year. The 30 mussels from each sampling were then allocated into three groups of 10. Two Cryptosporidium spp. genes (18S and GP60) were targeted for DNA amplification from the samples obtained. After purification, all of the products obtained were sequenced and phylogenetic analyses were performed. Of the 72 samples analyzed using the nested-PCR for the 18S gene target, 29.2% were positive for the presence of Cryptosporidium spp. Of these samples, 52.4% were collected at site A (ie 11/21) and 47.6% at site B (ie 10/21). The 18S genes of all the samples considered positive for Cryptosporidium spp. were sequenced, and the following three species were identified: Cryptosporidium parvum, C. meleagridis, and C. andersoni. Three distinct C. parvum subtypes (IIaA19G2R2; IIaA20G2R2; IIaA20G3R2) were identified using the GP60 gene. More studies to evaluate the zoonotic potential of this species should be performed as both sampling locations contain human and/or animal fecal contaminants.
诸如动物粪便径流、有机肥料施用以及部分处理或未处理污水的排放等污染源，可能导致隐孢子虫属对水生环境造成污染。贻贝作为食物的质量与发现这些双壳贝类的海洋环境的卫生状况密切相关。海洋软体动物是滤食性动物，能够在其组织中保留隐孢子虫卵囊，因此起到生物指示物的作用。在巴西里约热内卢州曼加拉蒂巴市的两个地点（A和B）共采集了72份合并的翡翠贻贝样本。采样工作包括在一年时间里每月从每个采集地点采集30只贻贝。然后将每个采样的30只贻贝分成三组，每组10只。针对从获得的样本中进行DNA扩增的目标是两个隐孢子虫属基因（18S和GP60）。纯化后，对所有获得的产物进行测序并进行系统发育分析。在使用巢式PCR针对18S基因靶点分析的72个样本中，29.2%的样本隐孢子虫属呈阳性。在这些样本中，52.4%是在A地点采集的（即21个样本中的11个），47.6%是在B地点采集的（即21个样本中的10个）。对所有被认为隐孢子虫属呈阳性的样本的18S基因进行了测序，并鉴定出以下三个物种：微小隐孢子虫、火鸡隐孢子虫和安氏隐孢子虫。使用GP60基因鉴定出了三种不同的微小隐孢子虫亚型（IIaA19G2R2；IIaA20G2R2；IIaA20G3R2）。由于两个采样地点都含有人类和/或动物粪便污染物，因此应该开展更多研究来评估该物种的人畜共患病潜力。
