Malkamäki Sanna, Näreaho Anu, Oksanen Antti, Sukura Antti
Department of Veterinary Biosciences, Faculty of Veterinary Medicine, University of Helsinki (FINPAR), Finland.
Department of Veterinary Biosciences, Faculty of Veterinary Medicine, University of Helsinki (FINPAR), Finland.
Parasitol Int. 2019 Jun;70:58-63. doi: 10.1016/j.parint.2019.01.008. Epub 2019 Jan 31.
Potential role of wild forest berries as a transmission vehicle for taeniid eggs was examined using non-zoonotic Taenia laticollis eggs as a model. The berries studied were bilberries (Vaccinium myrtillus) (1 m plot, n = 10) and lingonberries (Vaccinium vitis-idaea) (1 m plot, n = 11). The plots in the managed forest were evenly sprayed with 30,000 or 60,000 T. laticollis eggs suspended in water, and berries were collected 24 h after spraying. The berries were rinsed with water, and the water was sieved through a 1-mm and a 63-μm sieve to remove coarse material and through a 20-μm sieve to collect possible eggs. A small proportion of the sieved material was examined by microscopy after treatment with fluorescent Calcofluor White stain, which binds to eggshell chitin. In the recovery tests in artificially spiked samples, the detection limit was 5 eggs in 100 g of commercial frozen bilberries and lingonberries. Taeniid eggs were detected in all of the 10 experimentally contaminated bilberry samples and in 10 of 11 lingonberry samples. The sieved debris was also analyzed for T. laticollis DNA using semi-quantitative PCR. All samples were positive in quantitative SYBR Green real-time PCR using a T. laticollis-specific primer pair amplifying a short fragment of mitochondrial NADH dehydrogenase subunit 1 gene. This indicates that forest berries contaminated in shrubs contained T. laticollis eggs, and that berries can serve as a vehicle for taeniid eggs and may pose a possible risk to humans.
以非人畜共患的宽颈绦虫(Taenia laticollis)虫卵为模型,研究了野生森林浆果作为绦虫卵传播媒介的潜在作用。所研究的浆果为欧洲越橘(Vaccinium myrtillus)(1米见方的地块,n = 10)和红果越橘(Vaccinium vitis-idaea)(1米见方的地块,n = 11)。在人工管理的森林地块中,均匀喷洒悬浮于水中的30000或60000个宽颈绦虫虫卵,喷洒24小时后采集浆果。浆果用水冲洗,冲洗水先后通过1毫米筛网和63微米筛网以去除粗颗粒物质,再通过20微米筛网以收集可能存在的虫卵。用荧光增白剂钙荧光白(Calcofluor White)染色处理后,通过显微镜检查一小部分过筛物质,该染色剂可与卵壳几丁质结合。在人工添加样本的回收率测试中,100克市售冷冻欧洲越橘和红果越橘中的检测限为5个虫卵。在所有10个经实验污染的欧洲越橘样本以及11个红果越橘样本中的10个样本中检测到了绦虫卵。还使用半定量PCR分析过筛后的残渣中是否存在宽颈绦虫DNA。使用扩增线粒体烟酰胺腺嘌呤二核苷酸脱氢酶亚基1基因短片段的宽颈绦虫特异性引物对,通过定量SYBR Green实时PCR检测,所有样本均呈阳性。这表明灌木丛中受污染的森林浆果含有宽颈绦虫虫卵,浆果可作为绦虫卵的传播媒介,可能对人类构成潜在风险。