Multiple identification of most important waterborne protozoa in surface water used for irrigation purposes by 18S rRNA amplicon-based metagenomics

Understanding waterborne protozoan parasites (WPPs) diversity has important implications in public health. In this study, we evaluated a NGS-based method as a detection approach to identify simultaneously most important WPPs using 18S rRNA high-throughput sequencing. A set of primers to target the V...

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Published inInternational journal of hygiene and environmental health Vol. 221; no. 1; pp. 102 - 111
Main Authors Moreno, Y., Moreno-Mesonero, L., Amorós, I., Pérez, R., Morillo, J.A., Alonso, J.L.
Format Journal Article
LanguageEnglish
Published Germany Elsevier GmbH 01.01.2018
Subjects
18S amplicon sequencing
Agricultural Irrigation
Cryptosporidium - isolation & purification
DNA Primers
Giardia - isolation & purification
Irrigation water
Metagenomics
Metagenomics - methods
Public health
RNA, Ribosomal, 18S - analysis
Water - parasitology
Waterborne protozoa parasites
18S amplicon sequencing
Metagenomics
Irrigation water
Public health
Waterborne protozoa parasites
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Summary:Understanding waterborne protozoan parasites (WPPs) diversity has important implications in public health. In this study, we evaluated a NGS-based method as a detection approach to identify simultaneously most important WPPs using 18S rRNA high-throughput sequencing. A set of primers to target the V4 18S rRNA region of WPPs such as Cryptosporidium spp., Giardia sp., Blastocystis sp., Entamoeba spp, Toxoplasma sp. and free-living amoebae (FLA) was designed. In order to optimize PCR conditions before sequencing, both a mock community with a defined composition of representative WPPs and a real water sample inoculated with specific WPPs DNA were prepared. Using the method proposed in this study, we have detected the presence of Giardia intestinalis, Acanthamoeba castellanii, Toxoplasma gondii, Entamoeba histolytica and Blastocystis sp. at species level in real irrigation water samples. Our results showed that untreated surface irrigation water in open fields can provide an important source of WPPs. Therefore, the methodology proposed in this study can establish a basis for an accurate and effective diagnostic of WPPs to provide a better understanding of the risk associated to irrigation water.
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ISSN:1438-4639
1618-131X
DOI:10.1016/j.ijheh.2017.10.008