Comparative analysis of the vaginal microbiome of pregnant women with either Trichomonas vaginalis or Chlamydia trachomatis

English version

Autoři: Simon Chengo Masha ^aff001; Collins Owuor ^aff001; Joyce Mwongeli Ngoi ^aff001; Piet Cools ^aff002; Eduard J. Sanders ^aff001; Mario Vaneechoutte ^aff002; Tania Crucitti ^aff007; Etienne P. de Villiers ^aff001
Působiště autorů: KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research-Coast, Kilifi, Kenya ^aff001; Laboratory for Bacteriology Research, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium ^aff002; Department of Biological Sciences, Pwani University, Kilifi, Kenya ^aff003; Centre for Tropical Medicine and Global Health, Nuffield Department of Medicine Research Building, University of Oxford, Oxford, United Kingdom ^aff004; Department of Public Health, Pwani University, Kilifi, Kenya ^aff005; Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United Kingdom ^aff006; HIV/STI Reference Laboratory, Department of Clinical Sciences, Institute of Tropical Medicine, Nationalestraat, Antwerp, Belgium ^aff007
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pone.0225545

Souhrn

Background

Although the significance of the human vaginal microbiome for health and disease is increasingly acknowledged, there is paucity of data on the differences in the composition of the vaginal microbiome upon infection with different sexually transmitted pathogens.

Method

The composition of the vaginal bacterial community of women with Trichomonas vaginalis (TV, N = 18) was compared to that of women with Chlamydia trachomatis (CT, N = 14), and to that of controls (N = 21) (women negative for TV, CT and bacterial vaginosis). The vaginal bacterial composition was determined using high throughput sequencing with the Ion 16S metagenomics kit of the variable regions 2, 4 and 8 of the bacterial 16S ribosomal RNA gene from the vaginal swab DNA extract of the women. QIIME and R package “Phyloseq” were used to assess the α- and β-diversity and absolute abundance of the 16S rRNA gene per sample in the three groups. Differences in taxa at various levels were determined using the independent T-test.

Results

A total of 545 operational taxonomic units (OTUs) were identified in all the three groups of which 488 occurred in all three groups (core OTUs). Bacterial α-diversity, by both Simpson’s and Shannon’s indices, was significantly higher, (p = 0.056) and (p = 0.001) respectively, among women with either TV or CT than among controls (mean α-diversity TV-infected > CT-infected > Controls). At the genus level, women infected with TV had a significantly (p < 0.01) higher abundance of Parvimonas and Prevotella species compared to both controls and CT-infected women, whereas women infected with CT had a significantly (p < 0.05) higher abundance of Anaerococcus, Collinsella, Corynebacterium and Dialister.

Conclusion

The vaginal microbiomes of TV and CT-infected women were markedly different from each other and from women without TV and CT. Future studies should determine whether the altered microbiomes are merely markers of disease, or whether they actively contribute to the pathology of the two genital infections.

Klíčová slova:

Microbiome – Trichomonas vaginalis – Chlamydia infection – Ribosomal RNA – Shannon index – Simpson index – Bacterial vaginosis – Prevotella infection

Zdroje

1. Sender R, Fuchs S, Milo R. Revised Estimates for the Number of Human and Bacteria Cells in the Body. PLoS biology. 2016;14(8):e1002533. Epub 2016/08/20. doi: 10.1371/journal.pbio.1002533 27541692; PubMed Central PMCID: PMC4991899.

2. Gill SR, Pop M, DeBoy RT, Eckburg PB, Turnbaugh PJ, Samuel BS, et al. Metagenomic Analysis of the Human Distal Gut Microbiome. Science (New York, NY). 2006;312(5778):1355–9. doi: 10.1126/science.1124234 PMC3027896. 16741115

3. Human Microbiome Project C. Structure, function and diversity of the healthy human microbiome. Nature. 2012;486(7402):207–14. Epub 2012/06/16. doi: 10.1038/nature11234 22699609; PubMed Central PMCID: PMC3564958.

4. Newman L, Rowley J, Vander Hoorn S, Wijesooriya NS, Unemo M, Low N, et al. Global Estimates of the Prevalence and Incidence of Four Curable Sexually Transmitted Infections in 2012 Based on Systematic Review and Global Reporting. PloS one. 2015;10(12):e0143304. Epub 2015/12/10. doi: 10.1371/journal.pone.0143304 26646541; PubMed Central PMCID: PMC4672879.

5. Alakomi HL, Skytta E, Saarela M, Mattila-Sandholm T, Latva-Kala K, Helander IM. Lactic acid permeabilizes gram-negative bacteria by disrupting the outer membrane. Applied and environmental microbiology. 2000;66(5):2001–5. Epub 2000/05/02. doi: 10.1128/aem.66.5.2001-2005.2000 10788373; PubMed Central PMCID: PMC101446.

6. Kaewsrichan J, Peeyananjarassri K, Kongprasertkit J. Selection and identification of anaerobic lactobacilli producing inhibitory compounds against vaginal pathogens. FEMS Immunology & Medical Microbiology. 2006;48(1):75–83. doi: 10.1111/j.1574-695X.2006.00124.x 16965354

7. Verhelst R, Verstraelen H, Claeys G, Verschraegen G, Van Simaey L, De Ganck C, et al. Comparison between Gram stain and culture for the characterization of vaginal microflora: Definition of a distinct grade that resembles grade I microflora and revised categorization of grade I microflora. BMC microbiology. 2005;5(1):61. doi: 10.1186/1471-2180-5-61 16225680

8. Antonio MA, Hawes SE, Hillier SL. The identification of vaginal Lactobacillus species and the demographic and microbiologic characteristics of women colonized by these species. The Journal of infectious diseases. 1999;180(6):1950–6. Epub 1999/11/24. doi: 10.1086/315109 10558952.

9. Vaneechoutte M. Lactobacillus iners, the unusual suspect. Research in microbiology. 2017;168(9–10):826–36. doi: 10.1016/j.resmic.2017.09.003 28951208.

10. Bell C, Hough E, Smith A, Greene L. Targeted screening for Trichomonas vaginalis in women, a pH-based approach. International journal of STD & AIDS. 2007;18(6):402–3. Epub 2007/07/05. doi: 10.1258/095646207781024892 17609030.

11. Martin DH, Zozaya M, Lillis RA, Myers L, Nsuami MJ, Ferris MJ. Unique Vaginal Microbiota That Includes an Unknown Mycoplasma-Like Organism Is Associated With Trichomonas vaginalis Infection. Journal of Infectious Diseases. 2013;207(12):1922–31. doi: 10.1093/infdis/jit100 23482642

12. Masha SC, Cools P, Descheemaeker P, Reynders M, Sanders EJ, Vaneechoutte M. Urogenital pathogens, associated with Trichomonas vaginalis, among pregnant women in Kilifi, Kenya: a nested case-control study. BMC Infect Dis. 2018;18(1):549. doi: 10.1186/s12879-018-3455-4 30400890; PubMed Central PMCID: PMC6219184.

13. Bautista CT, Wurapa EK, Sateren WB, Morris SM, Hollingsworth BP, Sanchez JL. Association of Bacterial Vaginosis With Chlamydia and Gonorrhea Among Women in the U.S. Army. American journal of preventive medicine. 2017;52(5):632–9. Epub 2016/11/07. doi: 10.1016/j.amepre.2016.09.016 27816380.

14. Larsson PG, Platz-Christensen JJ, Dalaker K, Eriksson K, Fahraeus L, Irminger K, et al. Treatment with 2% clindamycin vaginal cream prior to first trimester surgical abortion to reduce signs of postoperative infection: a prospective, double-blinded, placebo-controlled, multicenter study. Acta obstetricia et gynecologica Scandinavica. 2000;79(5):390–6. Epub 2000/06/01. 10830767.

15. Ralph SG, Rutherford AJ, Wilson JD. Influence of bacterial vaginosis on conception and miscarriage in the first trimester: cohort study. Bmj. 1999;319(7204):220–3. Epub 1999/07/23. doi: 10.1136/bmj.319.7204.220 10417083; PubMed Central PMCID: PMC28171.

16. Llahi-Camp JM, Rai R, Ison C, Regan L, Taylor-Robinson D. Association of bacterial vaginosis with a history of second trimester miscarriage. Hum Reprod. 1996;11(7):1575–8. Epub 1996/07/01. doi: 10.1093/oxfordjournals.humrep.a019440 8671507.

17. Parry S, Strauss JF 3rd. Premature rupture of the fetal membranes. N Engl J Med. 1998;338(10):663–70. Epub 1998/03/05. doi: 10.1056/NEJM199803053381006 9486996.

18. Leitich H, Bodner-Adler B, Brunbauer M, Kaider A, Egarter C, Husslein P. Bacterial vaginosis as a risk factor for preterm delivery: a meta-analysis. American journal of obstetrics and gynecology. 2003;189. doi: 10.1067/mob.2003.339 12861153

19. Liu L, Johnson HL, Cousens S, Perin J, Scott S, Lawn JE, et al. Global, regional, and national causes of child mortality: an updated systematic analysis for 2010 with time trends since 2000. Lancet. 2012;379(9832):2151–61. Epub 2012/05/15. doi: 10.1016/S0140-6736(12)60560-1 22579125.

20. Masha SC, Wahome E, Vaneechoutte M, Cools P, Crucitti T, Sanders EJ. High prevalence of curable sexually transmitted infections among pregnant women in a rural county hospital in Kilifi, Kenya. PloS one. 2017;12(3):e0175166. doi: 10.1371/journal.pone.0175166 28362869.

21. Nugent RP, Krohn MA, Hillier SL. Reliability of diagnosing bacterial vaginosis is improved by a standardized method of gram stain interpretation. Journal of clinical microbiology. 1991;29(2):297–301. 1706728; PubMed Central PMCID: PMC269757.

22. Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, et al. QIIME allows analysis of high-throughput community sequencing data. Nature methods. 2010;7(5):335–6. Epub 2010/04/13. doi: 10.1038/nmeth.f.303 20383131; PubMed Central PMCID: PMC3156573.

23. DeSantis TZ, Hugenholtz P, Larsen N, Rojas M, Brodie EL, Keller K, et al. Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB. Applied and environmental microbiology. 2006;72(7):5069–72. doi: 10.1128/AEM.03006-05 16820507; PubMed Central PMCID: PMC1489311.

24. Zakrzewski M, Proietti C, Ellis JJ, Hasan S, Brion MJ, Berger B, et al. Calypso: a user-friendly web-server for mining and visualizing microbiome-environment interactions. Bioinformatics (Oxford, England). 2017;33(5):782–3. Epub 2016/12/28. doi: 10.1093/bioinformatics/btw725 28025202; PubMed Central PMCID: PMC5408814.

25. Brotman RM, Bradford LL, Conrad M, Gajer P, Ault K, Peralta L, et al. Association between Trichomonas vaginalis and vaginal bacterial community composition among reproductive-age women. Sex Transm Dis. 2012;39(10):807–12. Epub 2012/09/26. doi: 10.1097/OLQ.0b013e3182631c79 23007708; PubMed Central PMCID: PMC3458234.

26. Srinivasan S, Fredricks DN. The human vaginal bacterial biota and bacterial vaginosis. Interdisciplinary perspectives on infectious diseases. 2008;2008:750479. Epub 2008/01/01. doi: 10.1155/2008/750479 19282975; PubMed Central PMCID: PMC2648628.

27. Tamarelle J, de Barbeyrac B, Le Hen I, Thiebaut A, Bebear C, Ravel J, et al. Vaginal microbiota composition and association with prevalent Chlamydia trachomatis infection: a cross-sectional study of young women attending a STI clinic in France. Sexually transmitted infections. 2018. Epub 2018/01/24. doi: 10.1136/sextrans-2017-053346 29358524.

28. Domann E, Hong G, Imirzalioglu C, Turschner S, Kuhle J, Watzel C, et al. Culture-independent identification of pathogenic bacteria and polymicrobial infections in the genitourinary tract of renal transplant recipients. Journal of clinical microbiology. 2003;41(12):5500–10. Epub 2003/12/10. doi: 10.1128/JCM.41.12.5500-5510.2003 14662931; PubMed Central PMCID: PMC309025.

29. Chen J, Wright K, Davis JM, Jeraldo P, Marietta EV, Murray J, et al. An expansion of rare lineage intestinal microbes characterizes rheumatoid arthritis. Genome medicine. 2016;8(1):43. Epub 2016/04/23. doi: 10.1186/s13073-016-0299-7 27102666; PubMed Central PMCID: PMC4840970.

30. Funke G, von Graevenitz A, Clarridge JE, Bernard KA. Clinical microbiology of coryneform bacteria. Clinical Microbiology Reviews. 1997;10(1):125–59. PMC172946. 8993861

31. Morio F, Jean-Pierre H, Dubreuil L, Jumas-Bilak E, Calvet L, Mercier G, et al. Antimicrobial susceptibilities and clinical sources of Dialister species. Antimicrob Agents Chemother. 2007;51(12):4498–501. Epub 2007/10/10. doi: 10.1128/AAC.00538-07 17923492; PubMed Central PMCID: PMC2167981.

32. Tito RY, Cypers H, Joossens M, Varkas G, Van Praet L, Glorieus E, et al. Brief Report: Dialister as a Microbial Marker of Disease Activity in Spondyloarthritis. Arthritis & rheumatology (Hoboken, NJ). 2017;69(1):114–21. Epub 2016/07/09. doi: 10.1002/art.39802 27390077.