Molecular Identification of Parasitic Protozoa <i>Sarcocystis</i> in Water Samples
2022
Živilė Strazdaitė-Žielienė | Agnė Baranauskaitė | Dalius Butkauskas | Elena Servienė | Petras Prakas
<i>Sarcocystis</i> parasites are among the most common parasitic protozoa in farm animals. So far, the diversity of these parasites has been mainly studied in animal carcasses by morphological or molecular methods. Research on parasitic protozoa in environmental samples is scarce due to the lack of an appropriate methodology and low concentrations of parasites. For these reasons, there is a paucity of validated methods for <i>Sarcocystis</i> identification from environmental samples. Therefore, the present study aims to investigate various molecular methods for <i>Sarcocystis</i> parasite identification in water samples. In the present study, the sample volume, sporocysts isolation, and various conventional PCR were evaluated, and species-specific primers for the identification of different <i>Sarcocystis</i> species have been developed. Of the methods studied, based on data the most appropriate method for the identification of analyzed <i>Sarcocystis</i> spp. in water bodies is nested PCR, using species-specific primers targeting the <i>cox1</i> gene. <i>Sarcocystis</i> DNA was detected in 111 out of 114 (97.4%) samples. This paper represents the first identification of <i>S</i>. <i>bovifelis</i>, <i>S</i>. <i>cruzi</i>, <i>S</i>. <i>hirsuta</i>, <i>S</i>. <i>arieticanis</i>, <i>S</i>. <i>tenella</i>, <i>S</i>. <i>capracanis</i>, <i>S</i>. <i>bertrami</i>, and <i>S</i>. <i>miescheriana</i> by PCR and sequencing in environmental water samples. Our pilot study is useful in developing techniques for the identification of <i>Sarcocystis</i> species from water samples.
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