Abstract
Several parasite species are shared between humans and pigs. We explored the application of next-generation sequencing-based metabarcoding supplemented with real-time PCR to fecal DNAs from 259 samples from 116 pigs in Denmark to detect and differentiate single-celled intestinal parasites of zoonotic relevance. Enterocytozoon bieneusi, Balantioides coli, and Giardia duodenalis were observed in 34/37 (92%), 148/259 (57%), and 86/259 (33%) samples, respectively. Entamoeba polecki ST1, E. polecki ST3, and Entamoeba hartmanni were detected in 104/259 (40%), 161/259 (62%), and 8/259 (3%) samples, respectively. Metabarcoding and real-time PCR detected Cryptosporidium in 90/259 (35%) and 239/259 (92%) of the samples, respectively, with Cryptosporidium suis and Cryptosporidium scrofarum observed in nearly equal proportions. Blastocystis subtypes 1, 3, 5, and 15 were found in 72 (28%), 6 (2%), 176 (68%), and 36 (14%) of 259 samples, respectively. Iodamoeba was identified in 1/259 samples (<1%), while none of 37 tested samples was positive for Dientamoeba fragilis. Our results illustrate how metabarcoding exemplifies a ‘one-fits-many’ approach to detecting intestinal single-celled parasites in feces supplemented with real-time PCR for selected parasites. Using metabarcoding with pathogen-specific assays may help detect emerging and previously underdetected pathogens and further elucidate the role of micro-eukaryotic parasites in human and animal health and disease.
Highlights
Several parasites can be hosted by both human and non-human hosts
We previously introduced a metabarcoding assay relying on amplicon-based next-generation sequencing (NGS) of nuclear ribosomal genes from bacteria, fungi, and parasites with automated software-based annotation of sequences to genus and—oftentimes—
The amplicon-based NGS-based approach to detecting and differentiating parasites used in this study has been applied in a number of studies involving human, animal, and environmental samples, including corneal scrapings, fecal samples, and sewage samples [14,15,22,33], and with different foci from clinically relevant opportunistic parasites to foodborne parasites [34]
Summary
Several parasites can be hosted by both human and non-human hosts. Zoonotic intestinal parasitic genera taking a toll on both human and animal health include Giardia and Cryptosporidium [1]. A few others can be observed with varying frequency in both humans and larger mammals, such as pigs, including Enterocytozoon, Balantioides, Blastocystis, Dientamoeba, and Entamoeba [2,3,4,5,6,7,8,9], the public health and animal health significance and epidemiology of which remain unclear. The screening of human fecal samples for parasites of clinical and epidemiological relevance is increasingly being assisted by DNA-based tools such as targeted conventional and real-time PCRs [10] and commercial solutions offering multiplex PCR assays. The results from the current routine approaches often indicate only the presence/absence of parasites, with little or no information on genetic diversity (such as species, lineages, genotypes, subtypes, etc.) due to their high morphological uniformity (e.g., Blastocystis or Giardia) or due to the use of pathogen-specific targeted assays
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
