Departamento de Medicina Veterinária Preventiva e Saúde Animal, FMVZ, USP, São Paulo, SP, Brazil.
Vet Parasitol. 2009 Dec 3;166(1-2):47-51. doi: 10.1016/j.vetpar.2009.07.033. Epub 2009 Jul 29.
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).
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。
