Abstract

Background & methodsBlastocystis sp. is one of the most prevalent unicellular eukaryote of the human large intestine in Chile and worldwide. It is classified in subtypes (STs), where using the polymorphic sequences of its 18S rRNA genes currently recognizes 22. STs 1–9 and ST12 have been reported in humans. It has been hypothesized that different STs of Blastocystis sp. differentially affect the clinical severity of the digestive disease in Irritable Bowel Syndrome (IBS) patients, but more studies ar4e needed to establish this statement. To contribute in the elucidation of the potential relationship between Blastocystis sp. subtypes and IBS severity, 37 IBS patient fecal samples were collected at hospitals in Santiago (Chile) and were screened for the presence of vacuolated forms of Blastocystis sp. by using conventional microscopy. Positive samples were submitted to PCR and sequencing for determining STs. The same procedure was performed in fecal samples from five non-IBS Blastocystis sp. carriers for preliminary comparative purpose. Results and discussionFour out of the 37 samples from the IBS patients were found positive for Blastocystis sp. (10.81%) by using microscopy. The presence of this microorganism in these four samples were confirmed by PCR and sequencing. Subtypes and their respective closest match alleles were searched and the ST1, ST2 and ST4 subtypes were found in these patients. ST4 subtype is scarcely detected in South America countries, being reported previously only in Colombia and Brazil. In this ST4 subtype we determined the allele 42 which is the most frequent allele observed in human Blastocystis isolates. In the non-IBS individuals' carriers, three subtypes were found: ST1, ST2 and ST3, even belonging to the same family group. Closest match alleles: 2, 12 and 34 here detected were also commonly reported globally. Instead of the small number of IBS patients studied here, the frequency of blastocystosis detected (10.81%) was lower than the prevalence of Blastocystis sp. infections described for the Chilean general population (30.4%). In Chile, clear correlation of Blastocystis sp. subtypes and IBS severity is still lacking with this study but it may lead and contribute to a better understanding of its pathogenicity and worldwide epidemiology.

Highlights

  • Blastocystis sp. inhabits the large intestine of humans and it is the most frequently observed enteric protist in Chile and throughout the world (Mercado and Schenone, 2004; Jiménez et al, 2019)

  • In order to investigate the putative involvement of Blastocystis sp. subtypes in Inflammatory Bowel Syndrome (IBS), we studied 37 IBS patients in Santiago, Chile for this protist, through its SSU rDNA subtyping and subtypes alleles

  • Blastocystis sp. subtypes ST1, ST2 and ST4 were observed in the samples from IBS patients, while ST1, ST2, ST3 were identified in the non-IBS samples (Table 1)

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Summary

Introduction

Blastocystis sp. inhabits the large intestine of humans and it is the most frequently observed enteric protist in Chile and throughout the world (Mercado and Schenone, 2004; Jiménez et al, 2019). Is subtyped through sequences of its small subunit ribosomal RNA (18S SSU rDNA) gene and classified as STs, being the most common worldwide the ST1, ST2 and ST3 subtypes (Stensvold et al, 2007; Seyer et al, 2016). Subtyping of this enteric protist has been performed in South America, mainly in Argentina, Bolivia, Brazil, Colombia, Ecuador and Peru (Ramirez et al, 2016) but not yet in Chile. Subtypes in IBS, we studied 37 IBS patients in Santiago, Chile for this protist, through its SSU rDNA subtyping and subtypes alleles. We determined STs and their respective alleles in fecal samples of five Chilean individuals with blastocystosis that included two children and three adult members of the same family, who had no IBS symptoms

Samples and patient characterization
Microscopy examination and molecular procedures
Results and discussion
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