俄罗斯联邦西北联邦区沃洛格达州养猪场中隐孢子虫的分子特征

A. Kryazhev, A. Novikov
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摘要

这些研究的目的是确定、识别、分析和研究俄罗斯联邦西北联邦区沃洛格达州养猪场中隐孢子虫的分布情况。在俄罗斯联邦,我们首次以西北联邦区沃洛格达州的各类养猪场为例,采用最新的分子遗传学方法,即对通过巢式 PCR 获得的 18S rRNA 基因片段的扩增子文库进行高通量测序,确定受检动物的所有年龄组都感染了斯科法氏隐孢子虫。 养猪场动物的感染率为 34%,农场动物的感染率为 32.4%。使用系统进化分析法分析 ASV 的分类归属,并在 GenBank 数据库中使用 blastn 算法进行补充分析,结果显示所有检测样本中总共存在 10 种 ASV 类型(扩增子序列变体),它们与作为 18S p 基因隐孢子虫 RNA 片段存入 GenBank 的序列具有高度相似性。 已确定的是,从葡萄牙、英国到中国、印度和澳大利亚等世界不同地理区域检测到的 ASV1 和 ASV2 类型在所有受检农场中均有发现,但数量明显不同。 其余 ASV 的数量要少得多,而且不会在不同农场之间重复出现。 这些序列可能属于梭状隐孢子虫亚种的地方种群。 值得注意的是,发现了 ASV8 型隐孢子虫属的独特序列,以后可将其描述为一个新物种。
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Molecular characteristics of Cryptosporidium scrofarum in pig farms of the Vologda region of the Northwestern Federal District of the Russian Federation
The purpose of these studies was to identify, identify, analyze and study the distribution of Cryptosporidium scrofarum in pigs in the  farms  of  the  Vologda  Region  of  the Northwestern Federal District of the Russian Federation.For the first time in the Russian Federation, using the example of the Vologda Oblast of the Northwestern Federal District in pig  farms  of  various  types  using  the  latest molecular  genetic  methods,  namely  using high-throughput  sequencing  of  amplicon libraries of fragments of the 18S rRNA gene obtained  because  of  nested  PCR,  we  have established the infection of C. scrofarum in all  age  groups  of  examined  animals.  The contamination of animals kept in pig farms was 34%, in farms - 32.4%. Animals that are fattened at the age of 13-24 weeks are most susceptible to infection.Analysis  of  ASV  taxonomic  affiliation using  phylogenetic  analysis,  supplemented by analysis using the blastn algorithm in the GenBank database, showed that a total of 10 ASV types (amplicon sequence variant) are present  in  all  tested  samples,  which  have high  similarity  to  sequences  deposited  in GenBank  as  fragments  of  the  18S  p  gene Cryptosporidium  scrofarum  RNA.  It  has been established that the types of ASV1 and ASV2  detected  in  various  geographical  regions of the world from Portugal and Great Britain  to  China,  India  and  Australia  are identified in all examined farms, although in significantly  different  quantities.  The  remaining ASVs are present in much smaller numbers  and  do  not  repeat  from  farm  to farm.  These  sequences  probably  belong  to local populations of subspecies Cryptosporidium scrofarum.  It  is  interesting  to  find  a unique sequence of the genus Cryptosporidium  of  type  ASV8,  which  can  later  be  described as a new species.
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