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Next-generation sequencing reveals wide genetic diversity of Blastocystis subtypes in chickens including potentially zoonotic subtypes

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Blastocystis sp. is an intestinal protist parasite commonly found in the feces of humans and animals worldwide. Blastocystis exhibits extensive genetic diversity and has been identified in humans and a variety of animals including other mammals and birds. Blastocystis subtypes do not exhibit strict host specificity which raises the possibility of zoonotic transmission through either direct contact or fecal contamination of food or water. However, reports detailing the subtypes and prevalence of Blastocystis in avian species are limited. Therefore, this study investigated the presence of Blastocystis in chickens by molecular characterization of the small subunit rRNA (SSU rRNA) gene. Fecal samples from 130 chickens were collected from local markets in Uberlândia and Belo Horizonte in the state of Minas Gerais, Brazil. To detect and identify subtypes of Blastocystis, a next-generation amplicon sequencing protocol was used. Forty-four of the 130 (33.8%) chickens examined were positive for Blastocystis. Blastocystis subtypes ST6 (23/130; 17.7%), ST7 (43/130; 33.1%), ST10 (1/130; 0.8%), ST14 (5/130; 3.8%), ST25 (1/130; 0.8%), and a novel subtype (ST29) (2/130; 1.5%) were observed. A nanopore sequencing strategy was used to obtain the near full-length SSU rRNA gene nucleotide sequence and validate novel subtype ST29. Mixed infections containing multiple subtypes were common and identified in 63.6% of Blastocystis-positive chickens. All positive samples contained one or both potentially zoonotic subtypes ST6 and ST7. The prevalence of Blastocystis in chickens was high, and molecular characterization mostly identified subtypes previously found in humans. Thus, chickens may be a source of human infection and environmental contamination.

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Availability of data and materials

Data generated in this study including all raw fastq files from Illumima MiSeq sequencing (deposited to the NCBI sequence read archive under the accession number SRR9872868-SRR9872911) and the nucleotide sequences (deposited in GenBank under the accession numbers MW538325-MW538478) have been made available at the time of publication.

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This study was supported by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), process number 99999.009537/2014-07 and by USDA-ARS Project No: 8042-32000-100-00-D.

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Correspondence to Monica Santin.

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This study was approved by the Ethics Committee in Animal Research of the Federal University of Uberlândia (CEUA-UFU).

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The authors declare no competing interests.

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Section Editor: Yaoyu Feng

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Maloney, J.G., da Cunha, M.J.R., Molokin, A. et al. Next-generation sequencing reveals wide genetic diversity of Blastocystis subtypes in chickens including potentially zoonotic subtypes. Parasitol Res 120, 2219–2231 (2021).

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