Fecal dysbiosis associated with colonic hypersensitivity and behavioral alterations in chronically Blastocystis-infected rats.
Animals
Area Under Curve
Behavior, Animal
/ physiology
Blastocystis
/ pathogenicity
Blastocystis Infections
/ complications
Colonic Diseases
/ complications
Disease Models, Animal
Dysbiosis
/ etiology
Fatty Acids, Volatile
/ analysis
Feces
/ microbiology
Microbiota
ROC Curve
Rats
Rats, Wistar
Serine Proteases
/ metabolism
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
04 06 2020
04 06 2020
Historique:
received:
22
10
2019
accepted:
11
04
2020
entrez:
6
6
2020
pubmed:
6
6
2020
medline:
15
12
2020
Statut:
epublish
Résumé
Infectious gastroenteritis is a risk factor for the development of post-infectious Irritable Bowel Syndrome (PI-IBS). Recent clinical studies reported a higher prevalence of the intestinal parasite Blastocystis in IBS patients. Using a rat model, we investigated the possible association between Blastocystis infection, colonic hypersensitivity (CHS), behavioral disturbances and gut microbiota changes. Rats were orally infected with Blastocystis subtype 4 (ST4) cysts, isolated from human stool samples. Colonic sensitivity was assessed by colorectal distension and animal behavior with an automatic behavior recognition system (PhenoTyper), the Elevated Plus Maze test and the Forced Swimming tests. Feces were collected at different time points after infection to study microbiota composition by 16 S rRNA amplicon sequencing and for short-chain fatty acid (SFCA) analysis. Blastocystis-infected animals had non-inflammatory CHS with increased serine protease activity. Infection was also associated with anxiety- and depressive-like behaviors. Analysis of fecal microbiota composition showed an increase in bacterial richness associated with altered microbiota composition. These changes included an increase in the relative abundance of Oscillospira and a decrease in Clostridium, which seem to be associated with lower levels of SCFAs in the feces from infected rats. Our findings suggest that experimental infection of rats with Blastocystis mimics IBS symptoms with the establishment of CHS related to microbiota and metabolic shifts.
Sections du résumé
BACKGROUND
Infectious gastroenteritis is a risk factor for the development of post-infectious Irritable Bowel Syndrome (PI-IBS). Recent clinical studies reported a higher prevalence of the intestinal parasite Blastocystis in IBS patients. Using a rat model, we investigated the possible association between Blastocystis infection, colonic hypersensitivity (CHS), behavioral disturbances and gut microbiota changes.
METHODS
Rats were orally infected with Blastocystis subtype 4 (ST4) cysts, isolated from human stool samples. Colonic sensitivity was assessed by colorectal distension and animal behavior with an automatic behavior recognition system (PhenoTyper), the Elevated Plus Maze test and the Forced Swimming tests. Feces were collected at different time points after infection to study microbiota composition by 16 S rRNA amplicon sequencing and for short-chain fatty acid (SFCA) analysis.
RESULTS
Blastocystis-infected animals had non-inflammatory CHS with increased serine protease activity. Infection was also associated with anxiety- and depressive-like behaviors. Analysis of fecal microbiota composition showed an increase in bacterial richness associated with altered microbiota composition. These changes included an increase in the relative abundance of Oscillospira and a decrease in Clostridium, which seem to be associated with lower levels of SCFAs in the feces from infected rats.
CONCLUSIONS
Our findings suggest that experimental infection of rats with Blastocystis mimics IBS symptoms with the establishment of CHS related to microbiota and metabolic shifts.
Identifiants
pubmed: 32499543
doi: 10.1038/s41598-020-66156-w
pii: 10.1038/s41598-020-66156-w
pmc: PMC7272397
doi:
Substances chimiques
Fatty Acids, Volatile
0
Serine Proteases
EC 3.4.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
9146Références
Johnson, A. C. & Greenwood-Van Meerveld, B. The Pharmacology of Visceral Pain. in. Advances in pharmacology (San Diego, Calif.) 75, 273–301 (2016).
Enck, P. et al. Irritable bowel syndrome. Nat. Rev. Dis. Prim. 2, 16014 (2016).
pubmed: 27159638
doi: 10.1038/nrdp.2016.14
Drossman, D. A. Functional gastrointestinal disorders: what’s new for Rome IV? Lancet Gastroenterol. Hepatol. 1, 6–8 (2016).
pubmed: 28404114
doi: 10.1016/S2468-1253(16)30022-X
Rodiño-Janeiro, B. K., Vicario, M., Alonso-Cotoner, C., Pascua-García, R. & Santos, J. A Review of Microbiota and Irritable Bowel Syndrome: Future in Therapies. Adv. Ther. 35, 289–310 (2018).
pubmed: 29498019
pmcid: 5859043
doi: 10.1007/s12325-018-0673-5
Fond, G. et al. Anxiety and depression comorbidities in irritable bowel syndrome (IBS): a systematic review and meta-analysis. Eur. Arch. Psychiatry Clin. Neurosci. 264, 651–60 (2014).
pubmed: 24705634
doi: 10.1007/s00406-014-0502-z
Lee, Y. Y., Annamalai, C. & Rao, S. S. C. Post-Infectious Irritable Bowel Syndrome. Curr. Gastroenterol. Rep. 19, 56 (2017).
pubmed: 28948467
doi: 10.1007/s11894-017-0595-4
Ohman, L. & Simrén, M. Pathogenesis of IBS: role of inflammation, immunity and neuroimmune interactions. Nat. Rev. Gastroenterol. Hepatol. 7, 163–73 (2010).
pubmed: 20101257
doi: 10.1038/nrgastro.2010.4
Bercík, P. et al. Visceral hyperalgesia and intestinal dysmotility in a mouse model of postinfective gut dysfunction. Gastroenterology 127, 179–87 (2004).
pubmed: 15236184
doi: 10.1053/j.gastro.2004.04.006
Beatty, J. K., Bhargava, A. & Buret, A. G. Post-infectious irritable bowel syndrome: mechanistic insights into chronic disturbances following enteric infection. World J. Gastroenterol. 20, 3976–85 (2014).
pubmed: 24744587
pmcid: 3983453
doi: 10.3748/wjg.v20.i14.3976
Downs, I. A., Aroniadis, O. C., Kelly, L. & Brandt, L. J. Postinfection Irritable Bowel Syndrome. J. Clin. Gastroenterol. 51, 869–877 (2017).
pubmed: 28885302
doi: 10.1097/MCG.0000000000000924
Klem, F. et al. Prevalence, Risk Factors, and Outcomes of Irritable Bowel Syndrome After Infectious Enteritis: A Systematic Review and Meta-analysis. Gastroenterology 152, 1042–1054.e1 (2017).
pubmed: 28069350
pmcid: 5367939
doi: 10.1053/j.gastro.2016.12.039
Tan, K. S. W. New Insights on Classification, Identification, and Clinical Relevance of Blastocystis spp. Clin. Microbiol. Rev. 21, 639–665 (2008).
pubmed: 18854485
pmcid: 2570156
doi: 10.1128/CMR.00022-08
Alfellani, M. A. et al. Variable geographic distribution of Blastocystis subtypes and its potential implications. Acta Trop. 126, 11–18 (2013).
pubmed: 23290980
doi: 10.1016/j.actatropica.2012.12.011
Rene, B. A., Stensvold, C. R., Badsberg, J. H. & Nielsen, H. V. Subtype analysis of Blastocystis isolates from Blastocystis cyst excreting patients. Am. J. Trop. Med. Hyg. 80, 588–92 (2009).
doi: 10.4269/ajtmh.2009.80.588
Ramírez, J. D. et al. Geographic distribution of human Blastocystis subtypes in South America. Infect. Genet. Evol. 41, 32–35 (2016).
pubmed: 27034056
doi: 10.1016/j.meegid.2016.03.017
Poirier, P. et al. Development and Evaluation of a Real-Time PCR Assay for Detection and Quantification of Blastocystis Parasites in Human Stool Samples: Prospective Study of Patients with Hematological Malignancies. J. Clin. Microbiol. 49, 975–983 (2011).
pubmed: 21177897
pmcid: 3067686
doi: 10.1128/JCM.01392-10
Domínguez-Márquez, M. V., Guna, R., Muñoz, C., Gómez-Muñoz, M. T. & Borrás, R. High prevalence of subtype 4 among isolates of Blastocystis hominis from symptomatic patients of a health district of Valencia (Spain). Parasitol. Res. 105, 949–955 (2009).
pubmed: 19471964
doi: 10.1007/s00436-009-1485-y
Olsen, K. E. P., Christiansen, D. B., Nielsen, H. V. & Stensvold, C. R. Blastocystis sp. Subtype 4 is Common in Danish Blastocystis-Positive Patients Presenting with Acute Diarrhea. Am. J. Trop. Med. Hyg. 84, 883–885 (2011).
pubmed: 21633023
pmcid: 3110361
doi: 10.4269/ajtmh.2011.11-0005
Tito, R. Y. et al. Population-level analysis of Blastocystis subtype prevalence and variation in the human gut microbiota. Gut gutjnl-2018-316106 (2018). https://doi.org/10.1136/gutjnl-2018-316106
pubmed: 30171064
pmcid: 6582744
doi: 10.1136/gutjnl-2018-316106
El Safadi, D. et al. Children of Senegal River Basin show the highest prevalence of Blastocystissp. ever observed worldwide. BMC Infect. Dis. 14, 164 (2014).
pubmed: 24666632
pmcid: 3987649
doi: 10.1186/1471-2334-14-164
Ragavan, N. D., Kumar, S., Chye, T. T., Mahadeva, S. & Shiaw-Hooi, H. Blastocystis sp. in Irritable Bowel Syndrome (IBS) - Detection in Stool Aspirates during Colonoscopy. Plos one 10, e0121173 (2015).
Nourrisson, C. et al. Blastocystis is associated with decrease of fecal microbiota protective bacteria: Comparative analysis between patients with irritable bowel syndrome and control subjects. Plos one 9 (2014).
pubmed: 25365580
pmcid: 4218853
doi: 10.1371/journal.pone.0111868
Rostami, A. et al. Erratum to: the role of Blastocystis sp. and Dientamoeba fragilis in irritable bowel syndrome: a systematic review and meta-analysis. Parasitol. Res. 116, 2611–2612 (2017).
pubmed: 28725935
doi: 10.1007/s00436-017-5556-1
Kesuma, Y., Firmansyah, A., Bardosono, S., Sari, I. P. & Kurniawan, A. Blastocystis ST-1 is associated with Irritable Bowel Syndrome-diarrhoea (IBS-D) in Indonesian adolescences. Parasite Epidemiol. Control 6, e00112 (2019).
pubmed: 31528737
pmcid: 6742775
doi: 10.1016/j.parepi.2019.e00112
Audebert, C. et al. Colonization with the enteric protozoa Blastocystis is associated with increased diversity of human gut bacterial microbiota. Sci. Rep. 6, 25255 (2016).
pubmed: 27147260
pmcid: 4857090
doi: 10.1038/srep25255
Nieves-Ramírez, M. E. et al. Asymptomatic Intestinal Colonization with Protist Blastocystis Is Strongly Associated with Distinct Microbiome Ecological Patterns. mSystems 3 (2018).
Rooks, M. G. & Garrett, W. S. Gut microbiota, metabolites and host immunity. Nat. Rev. Immunol. 16, 341–352 (2016).
pubmed: 27231050
pmcid: 5541232
doi: 10.1038/nri.2016.42
Defaye, M. et al. Efficient and reproducible experimental infections of rats with Blastocystis spp. Plos one 13, e0207669 (2018).
pubmed: 30452467
pmcid: 6242359
doi: 10.1371/journal.pone.0207669
Cekin, A. H. et al. Blastocystosis in patients with gastrointestinal symptoms: a case–control study. BMC Gastroenterol. 12, 122 (2012).
pubmed: 22963003
pmcid: 3444885
doi: 10.1186/1471-230X-12-122
Stark, D., van Hal, S., Marriott, D., Ellis, J. & Harkness, J. Irritable bowel syndrome: a review on the role of intestinal protozoa and the importance of their detection and diagnosis. Int. J. Parasitol. 37, 11–20 (2007).
pubmed: 17070814
doi: 10.1016/j.ijpara.2006.09.009
Toro Monjaraz, E. M. et al. Blastocystis Hominis and Chronic Abdominal Pain in Children: Is there an Association between Them? J. Trop. Pediatr. 64, 279–283 (2018).
pubmed: 28985427
doi: 10.1093/tropej/fmx060
Salvador, F. et al. Epidemiological and clinical profile of adult patients with Blastocystis sp. infection in Barcelona, Spain. Parasit. Vectors 9, 548 (2016).
pubmed: 27741951
pmcid: 5064914
doi: 10.1186/s13071-016-1827-4
Poirier, P., Wawrzyniak, I., Vivarès, C. P., Delbac, F. & El Alaoui, H. New insights into Blastocystis spp.: A potential link with irritable bowel syndrome. PLoS Pathog. 8, 1–4 (2012).
doi: 10.1371/journal.ppat.1002545
Farzaei, M. H., Bahramsoltani, R., Abdollahi, M. & Rahimi, R. The Role of Visceral Hypersensitivity in Irritable Bowel Syndrome: Pharmacological Targets and Novel Treatments. J. Neurogastroenterol. Motil. 22, 558–574 (2016).
pubmed: 27431236
pmcid: 5056566
doi: 10.5056/jnm16001
Leder, K., Hellard, M. E., Sinclair, M. I., Fairley, C. K. & Wolfe, R. No correlation between clinical symptoms and Blastocystis hominis in immunocompetent individuals. J. Gastroenterol. Hepatol. 20, 1390–1394 (2005).
pubmed: 16105126
doi: 10.1111/j.1440-1746.2005.03868.x
Puthia, M. K., Lu, J. & Tan, K. S. W. Blastocystis ratti contains cysteine proteases that mediate interleukin-8 response from human intestinal epithelial cells in an NF-kappaB-dependent manner. Eukaryot. Cell 7, 435–43 (2008).
pubmed: 18156286
doi: 10.1128/EC.00371-07
Lim, M. X. et al. Differential regulation of proinflammatory cytokine expression by mitogen-activated protein kinases in macrophages in response to intestinal parasite infection. Infect. Immun. 82, 4789–801 (2014).
pubmed: 25156742
pmcid: 4249314
doi: 10.1128/IAI.02279-14
Camilleri, M. et al. Intestinal barrier function in health and gastrointestinal disease. Neurogastroenterol. Motil. 24, 503–512 (2012).
pubmed: 22583600
pmcid: 5595063
doi: 10.1111/j.1365-2982.2012.01921.x
Dagci, H. et al. Epidemiological and diagnostic features of blastocystis infection in symptomatic patients in izmir province, Turkey. Iran. J. Parasitol. 9, 519–29
Nourrisson, C. et al. On Blastocystis secreted cysteine proteases: a legumain-activated cathepsin B increases paracellular permeability of intestinal Caco-2 cell monolayers. Parasitology 143, 1713–1722 (2016).
pubmed: 27609526
doi: 10.1017/S0031182016001396
Saitou, M. et al. Complex phenotype of mice lacking occludin, a component of tight junction strands. Mol. Biol. Cell 11, 4131–42 (2000).
pubmed: 11102513
pmcid: 15062
doi: 10.1091/mbc.11.12.4131
Hussein, E. M., Hussein, A. M., Eida, M. M. & Atwa, M. M. Pathophysiological variability of different genotypes of human Blastocystis hominis Egyptian isolates in experimentally infected rats. Parasitol. Res. 102, 853–860 (2008).
pubmed: 18193282
doi: 10.1007/s00436-007-0833-z
Denoeud, F. et al. Genome sequence of the stramenopile Blastocystis, a human anaerobic parasite. Genome Biol. 12, R29 (2011).
pubmed: 21439036
pmcid: 3129679
doi: 10.1186/gb-2011-12-3-r29
Gentekaki, E. et al. Extreme genome diversity in the hyper-prevalent parasitic eukaryote Blastocystis. Plos Biol. 15, e2003769 (2017).
pubmed: 28892507
pmcid: 5608401
doi: 10.1371/journal.pbio.2003769
Cenac, N. et al. Role for protease activity in visceral pain in irritable bowel syndrome. J. Clin. Invest. 117, 636–647 (2007).
pubmed: 17304351
pmcid: 1794118
doi: 10.1172/JCI29255
Róka, R. et al. A pilot study of fecal serine-protease activity: a pathophysiologic factor in diarrhea-predominant irritable bowel syndrome. Clin. Gastroenterol. Hepatol. 5, 550–5 (2007).
pubmed: 17336590
doi: 10.1016/j.cgh.2006.12.004
Gecse, K. et al. Increased faecal serine protease activity in diarrhoeic IBS patients: a colonic lumenal factor impairing colonic permeability and sensitivity. Gut 57, 591–599 (2008).
pubmed: 18194983
doi: 10.1136/gut.2007.140210
Ibeakanma, C. et al. Brain–Gut Interactions Increase Peripheral Nociceptive Signaling in Mice With Postinfectious Irritable Bowel Syndrome. Gastroenterology 141, 2098–2108.e5 (2011).
pubmed: 21856270
doi: 10.1053/j.gastro.2011.08.006
Tooth, D. et al. Characterisation of faecal protease activity in irritable bowel syndrome with diarrhoea: origin and effect of gut transit. Gut 63, 753–60 (2014).
pubmed: 23911555
doi: 10.1136/gutjnl-2012-304042
Steck, N., Mueller, K., Schemann, M. & Haller, D. Bacterial proteases in IBD and IBS. Gut 61, 1610–1618 (2012).
pubmed: 21900548
doi: 10.1136/gutjnl-2011-300775
Shah, E., Rezaie, A., Riddle, M. & Pimentel, M. Psychological disorders in gastrointestinal disease: epiphenomenon, cause or consequence? Ann. Gastroenterol. 27, 224–230 (2014).
pubmed: 24974805
pmcid: 4073018
Rosa, P. B. et al. Folic acid prevents depressive-like behavior induced by chronic corticosterone treatment in mice. Pharmacol. Biochem. Behav. 127, 1–6 (2014).
pubmed: 25316305
doi: 10.1016/j.pbb.2014.10.003
Crumeyrolle-Arias, M. et al. Absence of the gut microbiota enhances anxiety-like behavior and neuroendocrine response to acute stress in rats. Psychoneuroendocrinology 42, 207–17 (2014).
pubmed: 24636517
doi: 10.1016/j.psyneuen.2014.01.014
Cho, H. et al. Voluntary movements as a possible non-reflexive pain assay. Mol. Pain 9, 25 (2013).
pubmed: 23688027
pmcid: 3716716
doi: 10.1016/j.jpain.2008.01.119
Yason, J. A., Liang, Y. R., Png, C. W., Zhang, Y. & Tan, K. S. W. Interactions between a pathogenic Blastocystis subtype and gut microbiota: in vitro and in vivo studies. Microbiome 7, 30 (2019).
pubmed: 30853028
pmcid: 6410515
doi: 10.1186/s40168-019-0644-3
X.-Y., Z. et al. Visceral hypersensitive rats share common dysbiosis features with irritable bowel syndrome patients. World J. Gastroenterol. 22, 5211–5227 (2016).
doi: 10.3748/wjg.v22.i22.5211
Zhao, K., Yu, L., Wang, X., He, Y. & Lu, B. Clostridium butyricum regulates visceral hypersensitivity of irritable bowel syndrome by inhibiting colonic mucous low grade inflammation through its action on NLRP6. Acta Biochim. Biophys. Sin. (Shanghai). 50, 216–223 (2018).
pubmed: 29329362
doi: 10.1093/abbs/gmx138
Hugenholtz, F. et al. Metatranscriptome analysis of the microbial fermentation of dietary milk proteins in the murine gut. Plos one 13, e0194066 (2018).
pubmed: 29664912
pmcid: 5903625
doi: 10.1371/journal.pone.0194066
Pozuelo, M. et al. Reduction of butyrate- and methane-producing microorganisms in patients with Irritable Bowel Syndrome. Sci. Rep. 5, 12693 (2015).
pubmed: 26239401
pmcid: 4523847
doi: 10.1038/srep12693
Sun, Q., Jia, Q., Song, L. & Duan, L. Alterations in fecal short-chain fatty acids in patients with irritable bowel syndrome: A systematic review and meta-analysis. Medicine (Baltimore). 98, e14513 (2019).
pubmed: 30762787
pmcid: 6408019
doi: 10.1097/MD.0000000000014513
Dalile, B., Van Oudenhove, L., Vervliet, B. & Verbeke, K. The role of short-chain fatty acids in microbiota–gut–brain communication. Nature Reviews Gastroenterology and Hepatology 16, 461–478 (2019).
pubmed: 31123355
doi: 10.1038/s41575-019-0157-3
Zimmermann, M. Ethical guidelines for investigations of experimental pain in conscious animals. Pain 16, 109–10 (1983).
pubmed: 6877845
doi: 10.1016/0304-3959(83)90201-4
Larauche, M., Gourcerol, G., Million, M., Adelson, D. W. & Taché, Y. Repeated psychological stress-induced alterations of visceral sensitivity and colonic motor functions in mice: influence of surgery and postoperative single housing on visceromotor responses. Stress 13, 343–54 (2010).
pubmed: 20536336
pmcid: 3295848
doi: 10.3109/10253891003664166
Aissouni, Y. et al. Acid-Sensing Ion Channel 1a in the amygdala is involved in pain and anxiety-related behaviours associated with arthritis. Sci. Rep. 7, 43617 (2017).
pubmed: 28321113
pmcid: 5340794
doi: 10.1038/srep43617
Slattery, D. A. & Cryan, J. F. Using the rat forced swim test to assess antidepressant-like activity in rodents. Nat. Protoc. 7, 1009–1014 (2012).
pubmed: 22555240
doi: 10.1038/nprot.2012.044
Tulstrup, M. V.-L. et al. Antibiotic Treatment Affects Intestinal Permeability and Gut Microbial Composition in Wistar Rats Dependent on Antibiotic Class. Plos one 10, e0144854 (2015).
pubmed: 26691591
pmcid: 4686753
doi: 10.1371/journal.pone.0144854
Caporaso, J. G. et al. QIIME allows analysis of high-throughput community sequencing data. Nat. Methods 7, 335–336 (2010).
pubmed: 20383131
pmcid: 3156573
doi: 10.1038/nmeth.f.303
Edgar, R. C. & Flyvbjerg, H. Error filtering, pair assembly and error correction for next-generation sequencing reads. Bioinformatics 31, 3476–3482 (2015).
pubmed: 26139637
doi: 10.1093/bioinformatics/btv401
Edgar, R. C. Search and clustering orders of magnitude faster than BLAST. Bioinformatics 26, 2460–2461 (2010).
pubmed: 20709691
doi: 10.1093/bioinformatics/btq461
pmcid: 20709691
Segata, N. et al. Metagenomic biomarker discovery and explanation. Genome Biol. 12 (2011).
pubmed: 21702898
pmcid: 3218848
doi: 10.1186/gb-2011-12-6-r60