Berto Bruno Pereira, Machado Evelyn Lina, Hossotani Camila Michele de Souza, Beretta Bruna Matarucco Sampaio, da Silva Débora Regina Romualdo, Nakamura Alex Akira, Meireles Marcelo Vasconcelos
Departamento de Biologia Animal, Instituto de Ciências Biológicas e da Saúde, Universidade Federal Rural do Rio de Janeiro, BR-465 km 7, Seropédica, Rio de Janeiro, 23897-000, Brazil.
Faculdade de Medicina Veterinária, Universidade Estadual Paulista, Rua Clóvis Pestana 793, Araçatuba, São Paulo, 16050-680, Brazil.
Syst Parasitol. 2023 Jun;100(3):245-259. doi: 10.1007/s11230-023-10084-6. Epub 2023 Jan 26.
Island canaries Serinus canaria (Linnaeus) are finches native to the North Atlantic Islands, however, they have a worldwide distribution in captivity due to their relevance as a pet bird. Coccidians are the most reported parasites of passerines worldwide, both in the wild and in captivity, being frequently associated with disease in passerines kept in rehabilitation centers and commercial breeders. This study aimed to identify coccidians from island canaries kept in captivity in Brazil. Three hundred and fifteen genomic DNA extracted from fecal samples of island canaries from different breeders from Southern and Southeastern Brazil were used to perform a nested PCR assay to amplify a partial fragment of the 28S small subunit ribosomal RNA gene (28S) of Isospora spp. Microscopic screening and morphological identification of Isospora oocysts was performed in fecal samples corresponding to PCR positive DNA samples. Fecal samples have been formalin-stored for approximately four years. Positivity rate for both microscopy and PCR was 10.5% (33/315). Posteriorly, Isospora serini (Aragão, 1933) Box, 1975 and Isospora canaria Box, 1975 were morphologically identified from fresh fecal samples of island canaries maintained by a breeder in the State of São Paulo, Southeastern Brazil, providing a genotypic characterization via sequencing of the mitochondrial cytochrome c oxidase subunit 1 (COI) and 28S genes. The 28S and COI sequences referring to the morphological identification of I. canaria was, respectively, 100% and 99% similar to sequences deposited as Isospora serinuse Yang, Brice, Elliot & Ryan, 2015 from island canaries kept in a rehabilitation center in Australia. The COI sequence referring to the morphological identification of I. serini was 100% similar to a sequence of an extraintestinal Isospora, corroborating this identification/sequencing since I. serini is the first isosporan with an extra-intestinal cycle demonstrated. The comparison of morphological and molecular data from I. canaria and I. serini from this study with published data of Isospora spp. from canaries worldwide, allowed the specific identification from preliminary generic identifications, correction of misidentifications, as well as the establishment of junior synonyms. Finally, this study provides morphological and molecular data that ensure the correct identification of the two Isospora spp. from island canaries in future studies worldwide.
岛金丝雀（Serinus canaria Linnaeus）是原产于北大西洋诸岛的雀类，然而，由于它们作为宠物鸟的重要性，在人工饲养环境中它们在全球范围内都有分布。球虫是全世界雀形目鸟类中报道最多的寄生虫，无论在野外还是人工饲养环境中，它们经常与康复中心和商业饲养者饲养的雀形目鸟类的疾病有关。本研究旨在鉴定巴西人工饲养的岛金丝雀体内的球虫。从巴西南部和东南部不同饲养者的岛金丝雀粪便样本中提取的315份基因组DNA，用于进行巢式PCR分析，以扩增等孢属（Isospora spp.）28S小亚基核糖体RNA基因（28S）的部分片段。对与PCR阳性DNA样本对应的粪便样本进行了等孢属卵囊的显微镜筛查和形态学鉴定。粪便样本已用福尔马林保存了大约四年。显微镜检查和PCR的阳性率均为10.5%（33/315）。随后，从巴西东南部圣保罗州一名饲养者饲养的岛金丝雀的新鲜粪便样本中，通过形态学鉴定出了塞氏等孢球虫（Isospora serini，Aragão，1933）Box，1975和加那利等孢球虫（Isospora canaria Box，1975），并通过对线粒体细胞色素c氧化酶亚基1（COI）和28S基因进行测序提供了基因型特征。与加那利等孢球虫形态学鉴定相关的28S和COI序列，分别与保存在澳大利亚一个康复中心的岛金丝雀的塞氏等孢球虫（Isospora serinuse Yang，Brice，Elliot & Ryan，2015）序列有100%和99%的相似性。与塞氏等孢球虫形态学鉴定相关的COI序列与一种肠外等孢属的序列100%相似，这证实了该鉴定/测序结果，因为塞氏等孢球虫是首个被证明具有肠外周期的等孢属寄生虫。将本研究中加那利等孢球虫和塞氏等孢球虫的形态学和分子数据与全球金丝雀等孢属的已发表数据进行比较，使得能够从初步的属级鉴定中进行种的鉴定、纠正错误鉴定以及确定次异名。最后，本研究提供了形态学和分子数据，确保在未来全球范围内的研究中能够正确鉴定岛金丝雀体内的这两种等孢属寄生虫。
