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Contribution of Blastocystishominis subtypes and associated inflammatory factors in development of irritable bowel syndrome

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Abstract

Blastocystis hominis with worldwide distribution is a human intestinal protozoa found in all countries. There have been differences in the severity of the pathogenesis of various Blastocystis spp. and a concomitant variation in the plasma concentration of the cytokines in patients with irritable bowel syndrome. In the present study, we aimed to demonstrate the contribution of B. hominis subtypes in the development of irritable bowel syndrome. Stool samples were collected from patients with gastrointestinal disorders. All samples were evaluated through native-lugol method. Total DNA was extracted. A PCR protocol was developed to amplify a specific region of the SSU ribosomal DNA (rDNA) gene. Serum levels of IL-6 and TNF-α were determined by immunoassay methods. The ClustalW algorithm was applied to align and blast the nucleotide sequences of the amplified region of the SSU rDNA gene. To evaluate the phylogenetic and molecular evolutionary of the nucleotide sequences, we used the MEGA software. In this study, we found 26 haplotypes of B. hominis in the studied samples which were collectively belong to five subtypes (ST1, ST2 in patients without irritable bowel syndrome vs. ST3 and two unknown subtypes in patients with irritable bowel syndrome). Result of ELISA showed a high level of IL-6 and TNF-α in the serum of patients with irritable bowel syndrome. The genetic heterogeneity of B. hominis and the existence of different subtypes of the protozoan in patients with IBS may shed light to the fact that some subtypes of parasites may involve in the pathogenesis of IBS.

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References

  • Abdulsalam AM, Ithoi I, Al-Mekhlafi HM, Al-Mekhlafi AM, Ahmed A, Surin J (2013) Subtype Distribution of Blastocystis Isolates in Sebha, Libya. PLoS One 8: e84372

  • Adl S, Simpson A, Farmer M, Andersen R, Anderson O, Barta J, Bowser S, Brugerolle G, Fensome R, Fredericq S, James T, Karpov S, Kugrens P, Krug J, Lane C, Lewis L, Lodge J, Lynn D, Mann D, Mc Court R, Mendoza L, Moestrup O, Mozley-Standridge S, Nerad T, Shearer C, Smirnov A, Spiegel F, Taylor M (2005) The new higher level classification of eukaryotes with emphasis on the taxonomy of protists. J Eukaryot Microbiol 52:399–451

    Article  PubMed  Google Scholar 

  • Arisue N, Hashimoto T, Yoshikawa H, Nakamura Y, Nakamura G, Nakamura F, Yano T, Hasegawa M (2002) Phylogenetic position of Blastocystis hominis and of stramenopiles inferred from multiple molecular sequence data. J Eukaryot Microbiol 49:42–53

    Article  CAS  PubMed  Google Scholar 

  • Awatif MA, Init I, Al-Mekhlafi HM, Al-Mekhlafi AM, Abdulhamid A, Johari S (2013) Subtype distribution of Blastocystis isolates in Sebha, Libya. PLoS One 8:e84372

    Article  Google Scholar 

  • Böhm-Gloning B, Knobloch J, Walderich B (1997) Five subgroups of Blastocystis hominis isolates from symptomatic and asymptomatic patients revealed by restriction site analysis of PCR-amplified 16S-like rDNA. Trop Med Int Health 2:771–778

    Article  PubMed  Google Scholar 

  • Clark CG (1997) Extensive genetic diversity in Blastocystis hominis. Mol Biochem Parasitol 87:79–86

    Article  CAS  PubMed  Google Scholar 

  • Coyle CM, Varughese J, Weiss LM, Tanowitz HB (2012) Blastocystis: to treat or not to treat. Clin Infect Dis 54:105

    Article  PubMed  Google Scholar 

  • Cruz LV, Plancarte CA, Alvarez MC, Valencia RS, Sásnchez RG, Vega FL (2003) Blastocystis hominis among food vendors in xochimilco markets. Rev Latinoam Microbiol 45:12–15

    Google Scholar 

  • Dieh S, Rincon M (2002) The two faces of IL-6 on Th1/Th2 differentiation. Mol Immunol 39:531–536

    Article  Google Scholar 

  • Eida AM, Eida MM (2008) Identification of Blastocystis hominis in patients with irritable bowel syndrome using microscopy and culture compared to PCR. Parasitol United J (PUJ) 1:87–92

    Google Scholar 

  • Eltayeb LB, Brair LS, Aljafari AS (2013) The impact of intestinal protozoan parasites among irritable bowel syndrome patients in Khartoum state. Al Neel Med J 3:47–57

    Google Scholar 

  • Heidari A, Rokni MB (2003) Prevalence of intestinal parasites among children in day-care centers in Damghan, Iran. Iran J Pub Health 32:31–34

    Google Scholar 

  • Hoevers J, Holman P, Logan K, Hommel M, Ashford R, Snowden K (2000) Restriction-fragment-length polymorphism analysis of small-subunit rRNA genes of Blastocystis hominis isolates from geographically diverse human hosts. Parasitol Res 86:57–61

    Article  CAS  PubMed  Google Scholar 

  • Hughes PA, Zola H, Penttila IA, Blackshaw LA, Andrews JM, Krumbiegel D (2013a) Immune activation in irritable bowel syndrome: can neuroimmune interactions explain symptoms? Am J Gasreroentrol 108:1066–1073

    Article  CAS  Google Scholar 

  • Hughes PA, Harrington AM, Castro J, Liebregts T, Adam B, Dallas JG, Isaacs NJ, Maldeniya L, Martin CM, Persson J, Andrews JM, Holtmann G, Blackshaw AL, Brierley SM (2013b) Sensory neuro-immune interactions differ between irritable bowel syndrome subtypes. Gut 62:1456–1465

    Article  CAS  PubMed  Google Scholar 

  • Ibeakanma C, Vanner S (2010) TNF alpha is a key mediator of the pronociceptive effects of mucosal supernatant from human ulcerative colitis on colonic DRG neurons. Gut 59:612–621

    Article  CAS  PubMed  Google Scholar 

  • Kaneda Y, Horiki N, Cheng XJ, Fujita Y, Maruyama M, Tachibana H (2001) Ribodemes of Blastocystis hominis isolated in Japan. Am J Trop Med Hyg 65:393–396

    CAS  PubMed  Google Scholar 

  • Khoshnood S, Rafiei A, Saki J, Alizadeh K (2015) Prevalence and genotype characterization of Blastocystis hominis among the baghmalek people in southwestern Iran in 2013–2014. Jundishapur J Microbiol 8:e23930

    Article  PubMed  PubMed Central  Google Scholar 

  • Liebregts T, Adam B, Bredack C, Röth A, Heinzel S, Lester S, Downie-Doyle S, Smith E, Drew P, Talley NJ, Holtmann G (2007) Immune activation in patients with irritable bowel syndrome. Gastroenterology 132:913–920

    Article  CAS  PubMed  Google Scholar 

  • Longstreth GF, Thompson WG, Chey WD, Houghton LA, Mearin F (2006) Functional bowel disorders. Gastroenterology 130:1480–1491

    Article  PubMed  Google Scholar 

  • Motazedian H, Gasemi H, Sadjjadi MS (2008) Genomic diversity of Blastocystis hominis from patients in southern Iran. Ann Trop Med Parasitol 102:85–88

    Article  CAS  PubMed  Google Scholar 

  • Nagel R, Bielefeldt-Ohmann H, Traub R (2014) Clinical pilot study: efficacy of triple antibiotic therapy in Blastocystis positive irritable bowel syndrome patients. Gut Pathogens 6:34

    PubMed  PubMed Central  Google Scholar 

  • Nourrisson C, Scanzi J, Pereira B, Nkoud Mongo C, Wawrzyniak I, Cian A, Viscogliosi E, Livrelli V, Delbac F, Dapoigny M, Poirier P (2014) Blastocystis is associated with decrease of fecal microbiota protective bacteria: comparative analysis between patients with irritable bowel syndrome and control subjects. PLoS One 9:e111868

    Article  PubMed  PubMed Central  Google Scholar 

  • O’Malley D, Liston M, Hyland NP, Dinan TG, Cryan JF (2011) Colonic soluble mediators from the maternal separation model of irritable bowel syndrome activate submucosal neurons via an interleukin-6-dependent mechanism. Am J Physiol Gastrointest Liver Physiol 300:G241–G252

    Article  PubMed  Google Scholar 

  • Pipatsatitpong D, Rangsin R, Leelayoova S, Naaglor T, Mungthin M (2012) Incidence and risk factors of Blastocystis infection in an orphanage in Bangkok. Parasite Vector 5:37

    Article  Google Scholar 

  • Poirier P, Wawrzyniak I, Vivare CP, Delbac F, El Alaoui H (2012) New insights into Blastocystis spp.: a potential link with irritable bowel syndrome. PLoS Pathogens 8:e1002545

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ramirez-Miranda ME, Hernandez-Castellanos R, Lopez-Escamilla E, Moncada D, Rodriguez-Magallan A, Pagaza-Melero C, Gonzalez-Angulo A, Flisser A, Kawa-Karasik S, Maravilla P (2010) Parasites in Mexican patients with irritable bowel syndrome: a case-control study. Parasite Vector 3:96

    Article  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  • Santín M, Gómez-Muñoz MT, Solano-Aguilar G, Fayer R (2011) Development of a new PCR protocol to detect and subtype Blastocystis spp. from humans and animals. Parasitol Res (2011) 109:205–212

  • Shawky A, Fouad A, Maha MA, Basyoni B, Reham A, Fahmy C, Mohamed H, Kobaisi D (2011) The pathogenic role of different Blastocystis hominis genotypes isolated from patients with irritable bowel syndrome. Arab J Gastroenterol 12:194–200

    Article  Google Scholar 

  • Singh RK, Pandey HP, Singh RH (2003) Irritable bowel syndrome: challenges ahead. Curr Sci 84:25

    Google Scholar 

  • Stensvold CR, Alfellani M, Clark G (2012) Levels of genetic diversity vary dramatically between Blastocystis subtypes Infection. Gen Evol 12:263–273

    Article  Google Scholar 

  • Tan KSW (2008) New insights on classification, identification and clinical relevance of Blastocystis spp. Clin Microbiol Rev 21:639–665

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tan KSW (2004) Blastocystis in humans and animals: new insights using modern methodologies. Vet Parasitol 9:121–144

    Article  Google Scholar 

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol and Evolution 24:1596–1599

  • Thathaisong U, Worapong J, Mungthin M, Tan-Ariya P, Viputtigul K, Sudatis A, Noonai A, Leelayoova S (2003) Blastocystis isolates from a pig and a horse are closely related to Blastocystis hominis. J Clin Microbiol 41:967–975

    Article  PubMed  PubMed Central  Google Scholar 

  • Vogelberg C, Stensvold CR, Monecke S, Ditzen A, Stopsack K, Heinrich-Grafe U, Pohlmann C (2010) Blastocystis sp. subtype 2 detection during recurrence of gastrointestinal and urticarial symptoms. Parasitol Int 59:469–471

    Article  PubMed  Google Scholar 

  • Wong KH, Ng GC, Lin RT, Yoshikawa H, Taylor MB, Tan KS (2008) Predominance of subtype 3 among Blastocystis isolates from a major hospital in Singapore. Parasitol Res 102:663–670

    Article  PubMed  Google Scholar 

  • Yakoob J, Jafri W, AsimBM AZ, NazSh IM, Khan R (2010) Blastocystis hominis and Dientamoeba fragilis in patients fulfilling irritable bowel syndrome criteria. Parasitol Res 107:679–684

    Article  PubMed  Google Scholar 

  • Yan Y, Su S, Lai R, Liao H, Ye J, Li X, Luo X, Chen G (2006) Genetic variability of Blastocystis hominis isolates in China. Parasitol Res 99:597–601

    Article  PubMed  Google Scholar 

  • Yoshikawa H, Abe N, Wu Z (2003) Genomic polymorphism among Blastocystis isolates and development of PCR-based identification of zoonotic isolates. J Eukaryot Microbiol 50:710–711

    Article  PubMed  Google Scholar 

  • Yoshikawa H, Abe N, Iwasawa M, Kitano S, Nagano I, Wu Z, Takahashi Y (2000) Genomic analysis of Blastocystis hominis strains isolated from two long term health care facilities. J Clin Microbiol 38:1324–1330

    CAS  PubMed  PubMed Central  Google Scholar 

  • Yoshikawa H, Wu Z, Kimata I, Iseki M, Ali IK, Hossain MB, Zaman V, Haque R, Takahashi Y (2004) Polymerase chain reaction-based genotype classification among human Blastocystis hominis populations isolated from different countries. Parasitol Res 92:22–29

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The work was supported by the Ilam University of Medical Scinences, Ilam, Iran, grant 908441 awarded to Dr. Asad Mirzaei. The authors would like to acknowledge the staff of Dr. Asadollahi laboratory for their kind assistance in the sampling.

Authors’ contributions

MA and BGH were involved in designing and supervising the fieldwork and in interpreting the data; AM, was involved in the collection of samples and molecular tests; MA supervised all genetic aspects in this study. MA and BGH wrote the manuscript. All authors have read and approved the final manuscript. MA is the guarantor of the paper.

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Authors and Affiliations

  1. Department of Parasitology, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran

    Marzieh Azizian, Gholam Basati, Ghobad Abangah & Asad Mirzaei

  2. Department of Biology, Islamic Azad University of Arak, Arak, Iran

    Marzieh Azizian

  3. Department of Microbiology, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran

    Mohammad Reza Mahmoudi

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  1. Marzieh Azizian
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  2. Gholam Basati
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  3. Ghobad Abangah
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  5. Asad Mirzaei
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Correspondence to Asad Mirzaei.

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The study was approved by the Ilam University of Medical Sciences Ethics Committee and registered with no: EC/92/H/132.

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The authors declare that they have no competing interests.

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Azizian, M., Basati, G., Abangah, G. et al. Contribution of Blastocystishominis subtypes and associated inflammatory factors in development of irritable bowel syndrome. Parasitol Res 115, 2003–2009 (2016). https://doi.org/10.1007/s00436-016-4942-4

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  • DOI: https://doi.org/10.1007/s00436-016-4942-4

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