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巴西圈养鸟类粪便样本中隐孢子虫属的分子特征。

Molecular characterization of Cryptosporidium spp. from fecal samples of birds kept in captivity in Brazil.

机构信息

Departamento de Medicina Veterinária Preventiva e Saúde Animal, FMVZ, USP, São Paulo, SP, Brazil.

出版信息

Vet Parasitol. 2009 Dec 3;166(1-2):47-51. doi: 10.1016/j.vetpar.2009.07.033. Epub 2009 Jul 29.

Abstract

The aim of this study was to determine the prevalence of Cryptosporidium species and genotypes in birds kept in captivity in Brazil. A total of 966 samples from 18 families of birds was collected and stored in 5% potassium dichromate solution at 4 degrees C until processing. Oocysts were purified in Sheather sugar solution following extraction of genomic DNA. Molecular analyses were performed using nested-PCR for amplification of fragments of the 18S subunit of rRNA gene and of the actin gene. Amplification of Cryptosporidium DNA fragments was obtained in 47 (4.86%) samples. Sequencing of amplified fragments and phylogenetic analyses allowed the identification of Cryptosporidium baileyi in a black vulture (Coragyps atratus), a domestic chicken (Gallus gallus domesticus) and a saffron finch (Sicalis flaveola); Cryptosporidium galli in canaries (Serinus canaria), a cockatiel (Nymphicus hollandicus) and lesser seed-finches (Oryzoborus angolensis); Cryptosporidium meleagridis in a domestic chicken (G. g. domesticus); Cryptosporidium parvum in a cockatiel (N. hollandicus); Cryptosporidium avian genotype I in a canary (S. canaria) and an Indian peafowl (Pavo cristatus); Cryptosporidium avian genotype II in ostriches (Struthio camelus) and Cryptosporidium avian genotype III in a cockatiel (N. hollandicus) and a peach-faced lovebird (Agapornis roseicolis).

摘要

本研究旨在确定巴西圈养鸟类中隐孢子虫种类和基因型的流行情况。共采集了 18 个鸟类科的 966 个样本,并储存在 4°C 的 5%重铬酸钾溶液中,直至处理。使用 Sheather 糖溶液提取基因组 DNA 后,对卵囊进行纯化。采用嵌套 PCR 对 18S 亚单位 rRNA 基因和肌动蛋白基因片段进行扩增,进行分子分析。在 47 个(4.86%)样本中获得了隐孢子虫 DNA 片段的扩增。扩增片段的测序和系统发育分析允许鉴定黑秃鹫(Coragyps atratus)、家鸡(Gallus gallus domesticus)和藏黄雀(Sicalis flaveola)中的隐孢子虫 baileyi;金丝雀(Serinus canaria)、玄凤鹦鹉(Nymphicus hollandicus)和小黍雀(Oryzoborus angolensis)中的隐孢子虫 galli;家鸡(G. g. domesticus)中的隐孢子虫 meleagridis;玄凤鹦鹉(N. hollandicus)中的隐孢子虫 parvum;金丝雀(S. canaria)和印度孔雀(Pavo cristatus)中的隐孢子虫禽基因型 I;鸵鸟(Struthio camelus)中的隐孢子虫禽基因型 II 和玄凤鹦鹉(N. hollandicus)和桃面爱情鸟(Agapornis roseicolis)中的隐孢子虫禽基因型 III。

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