M.A. KAGANOVA, N.V. SPIRIDONOVA, V.O. KATYUSHINA

Samara State Medical University, Samara

 Contact details:

Kaganova M.A. – PhD (medicine), Associate Professor of the Department of Obstetrics and Gynecology

Address: 80 Polevaya St., Russian Federation, Samara, 443100, tel.: +7-846-207-19-68, e-mail: [email protected]

The purpose — to study the microbial placental landscape at premature rupture of membrane (PROMt) in comparison with intact membranes at full-term pregnancy.

Material and methods. At the City Clinical Hospital No. 1 named after N.I. Pirogov, 43 pregnant women at 37–41 weeks of gestation (24 persons with PROMt in the main group and 19 patients in the control group with intact membranes) placental tissue were taken during elective cesarean section for RT-PCR of the following microorganisms: Lactobacillus spp, Enterobacteriaceae, Streptococcus spp., Staphylococcus spp., Gardnerellavaginalis / Prevotella bivia / Porphyromonas spp., Eubacterium spp., Sneathia spp. / Leptotrihia spp. / Fusobacteriumspp, Megasphaera spp. / Veillonella spp. / Dialister spp., Lachnobacterium spp. / Clostridium spp., Mobiluncus spp. / Corynebacterium spp., Peptostreptococcus spp., Atopobiumvaginae, Mycoplasma hominis, Ureaplasma (urealyticum + parvum), Candida spp., Mycoplasma genitalium.

Results. The presence of a total bacterial mass of 10^3.1–10^4.5 Ge/sample was revealed at the placenta of full-term pregnancy. Sterile placentas were found both in the control (21,1%) and in the main groups (41,6%). In the placenta samples, we observed the predominance of the laboratory total bacterial mass (TBM) over the calculated one, i.e. they contained DNA of microorganisms that was not identified by the standard panel — the «unidentifiable» microorganisms (in the main group — 25,0% and in the control group — 52,6%). Enterobacteriaceae spp. (102,6 Ge/sample) were detected on the placental tissues of the patients with intact membrane. The placental microflora is gradually replaced by the vaginal one, obligate and facultative microflora of the vaginal biotope appeared. The presence of Lactobacillus spp. in placental tissues is characteristic only for patients with PROMt.

Conclusion. A small amount of bacterial mass can be detected by RT-PCR at the physiologically full-term pregnancy, represented by Enterobacteriaceae spp. Lactobacillus spp. and anaerobic vaginal flora are also detected on placental samples at full-term pregnancy with PROMt.

Key words: premature rupture of membrane, femoflor, placenta, polymerase chain reaction, microbiota.

REFERENCES

1. Han Y.W., Shen T., Chung P., Buhimschi I.A., Buhimschi C.C. Uncultivated bacteria as etiologic agents of intra-amniotic inflammation leading to preterm birth. J Clin Microbiol, 2009, vol. 47, pp. 38–47. DOI: 10.1128/JCM.01206-08
2. Goldenberg R.L., Hauth J.C., Andrews W.W. Intrauterine infection and preterm delivery. N Engl J Med, 2000, vol. 342, pp. 1500–1507. DOI: 10.1056/NEJM200005183422007
3. Stout M.J., Conlon B., Landeau M., Lee I., Bower C., Zhao Q., Roehl K.A., Nelson D.M., Macones G.A., Mysorekar I.U. Identification of intracellular bacteria in the basal plate of the human placenta in term and preterm gestations. Am J Obstet Gynecol, 2013, vol. 208: 226 e221–7.
4. Satokari R., Gronroos T., Laitinen K., Salminen S. Isolauri E. Bifidobacterium and Lactobacillus DNA in the human placenta. Lett Appl Microbiol, 2009, vol. 48, pp. 8–12.
5. Parnell L.A., Briggs C.M., Cao B., Delannoy-Bruno O., Schrieffer A.E., Mysorekar I.U. Microbial communities in placentas from term normal pregnancy exhibit spatially variable profiles. Sci Rep, 2017, vol. 7, p. 11200. DOI: 10.1038/s41598-017-11514-4
6. Lim et al. Microbiome, 2018, vol. 6, p. 87. https://doi.org/10.1186/s40168-018-0475-7
7. Collado M.C., Rautava S., Aakko J., Isolauri E., Salminen S. Human gut colonisation may be initiated in utero by distinct microbial communities in the placenta and amniotic fluid. Sci Rep, 2016, vol. 6, p. 23129.
8. Aagaard K., Ma J., Antony K.M., Ganu R., Petrosino J., Versalovic J. The placenta harbors a unique microbiome. SciT ransl Med, 2014, vol. 6, p. 237ra265. Caroline M. Mitchell,
9. Anoria9, available at: https://www.ncbi.nlm.nih.gov/pubmed/?term=HAICK%20A%5BAuthor%5D9.&9.cauthor=true9.&9.cauthor_uid=25524398″9         Kathy Agnew.           David N. Fredricks.       David Eschenbach. Colonization of the upper genital tract by vaginal bacterial species in non-pregnant women. Am J Obstet.     Published online, 2014, Dec 16. DOI: 9.10.1016/j.ajog.2014.11.043PMCID: PMC4754962 NIHMSID: NIHMS650669 PMID: 9.         25524398.

10. Chen H.J., Gur T.L. Intrauterine Microbiota: Missing, or the Missing Link? Trends Neurosci. 2019, vol. 42, pp. 402–413.
11. Prince A.L., Ma J., Kannan P.S., Alvarez M., Gisslen T., Harris R.A., Sweeney E.L., Knox C.L., Lambers D.S., Jobe A.H. et al. The placental membrane microbiome is altered among subjects with spontaneous preterm birth with and without chorioamnionitis. Am J Obstet Gynecol, 2016, vol. 214.
12. Popova E.N., Gordeev I.G. Modern ideas about human microbiota. MICROBIOT. Moscow, 2019. Pp. 5–19.
13. Fox C., Eichelberger K. Maternal microbiome and pregnancy outcomes. Fertil Steril., 2015, vol. 104 (6), pp. 1358–1363.
14. Peterson J., Garges S., Giovanni M., McInnes P., Wang L., Schloss J.A., Bonazzi V., McEwen J.E., Wetterstrand K.A. et al. The NIH Human Microbiome Project. Genome Res, 2009, vol. 19, pp. 2317–2323.
15. Lipatov I.S., Tezikov Yu.V., Martynova N.V., Mingalieva L.K., Gogel’ L.Yu., Belokoneva T.S., Kalinkina O.B., Zhernakova E.V., Yusupova R.R. A universal approach to the prevention of pathological pregnancy syndrome. Nauka i innovatsii v meditsine, 2017, vol. 1 (5), pp. 13–23 (in Russ.).
16. Boldyreva M.N., Lipova E.V., Alekseev L.P., Vitvitskaya Yu.G., Gus’kova I.A. Characterization of the biota of the urogenital tract in women of reproductive age by real-time PCR. Zhurnal akusherstva i zhenskikh bolezney, 2009, vol. LVIII, no. 6, pp. 36–42 (in Russ.).
17. Sukhikh G.T., Prilepskaya V.N., Trofimov D.Yu., Donnikov A.E., Aylamazyan E.K. et al. Primenenie metoda polimeraznoy tsepnoy reaktsii v real’nom vremeni dlya otsenki mikrobiotsenoza urogenital’nogo trakta u zhenshchin (test femoflor): meditsinskaya tekhnologiya [Application of the real-time polymerase chain reaction method to assess the microbiocenosis of the urogenital tract in women (femoflor test): medical technology]. Moscow, 2011. P. 36.
18. Voroshilina E.S., Tumbinskaya L.V., Donnikov A.E., Plotko E.E., Khayutin L.V. Vaginal biocenosis in terms of quantitative polymerase chain reaction: what is the norm? Akusherstvo i ginekologiya, 2011, pp. 57–65 (in Russ.).
19. Kaganova M.A., Spiridonova N.V., Kazakova A.V., Devyatova O.O., Galkina D.A., Golovina O.N. Features of the microbiota of the cervical canal during prenatal rupture of amniotic fluid and full-term pregnancy. Akusherstvo i ginekologiya, 2019, no. 5, pp. 77–84 (in Russ.). https://dx.doi.org/10.18565/aig.2019.5.77-84
20. Lauder A.P., Roche A.M., Sherrill-Mix S., Bailey A., Laughlin A.L., Bittinger K. et al. Comparison of placenta samples with contamination controls does not provide evidence for a distinct placenta microbiota. Microbiome, 2014, vol. 4, p. 29. DOI: 10.1186/s40168-016-0172-3
21. Kevin R. Theis, Roberto Romero, Andrew D. Winters, Jonathan M. Greenberg, Nardhy Gomez-Lopez, Ali Alhousseini, Janine Bieda, Eli Maymon, Percy Pacora, Jennifer M. Fettweis, Gregory A. Buck, Kimberly K. Jeferson, Jerome F. Strauss, Offer Erez, Sonia S. Hassan. Does the human placenta delivered at term have a microbiota? Results of cultivation, quantitative real-time PCR, 16S rRNA gene sequencing, and metagenomics. J Matern Fetal Neonatal Med, 2019, Dec, vol. 29, pp. 1–16.
22. de Goffau M.C., Lager S., Sovio U., Gaccioli F., Cook E., Peacock S.J. et al. Human placenta has no microbiome but can contain potential pathogens. Nature, 2019, vol. 572, pp. 1–21. DOI: 10.1038/s41586-019-1451-5
23. Kevin R. Theis, Roberto Romero, Jonathan M. Greenberg, Andrew D. Winters, Valeria Garcia-Flores, KenichiroMotomura, Madison M. Ahmad, Jose Galaz, Marcia Arenas-Hernandez, Nardhy Gomez-Lopeza. No Consistent Evidence for Microbiota in Murine Placental and Fetal Tissues. mSphere, Feb, 2020, vol. 5 (1), p. e00933-19. DOI: 10.1128/mSphere.00933-19
24. Satokari R., Gronroos T., Laitinen K., Salminen S., Isolauri E. Bifidobacterium and Lactobacillus DNA in the human placenta. Lett Appl Microbiol, 2009, vol. 48, pp. 8–12.
25. Lihong Zhu, Fei Luo, Wenjing Hu, Yang Han, Yuezhu Wang, Huajun Zheng, Xiaokui Guo, Jinhong Qin. Bacterial Communities in the Womb During Healthy Pregnancy. Front. Microbiol, Sept. 2018. DOI.org/10.3389/fmicb.2018.02163
26. Bagga R., Arora P. Genital Micro-Organisms in Pregnancy. Front. Public Health, vol. 8, p. 225. DOI: 10.3389/fpubh. 2020.00225
27. Martinez K.A., Romano-Keeler J., Zackular J.P., Moore D.J., Brucker R.M., Hooper C., Meng S., Brown N., Mallal S., Reese J. et al. Bacterial DNA is present in the fetal intestine and overlaps with that in the placenta in mice. PLoS ONE, 2018, vol. 13, p. e0197439.
28. Parnell L.A., Briggs C.M., Cao B., Delannoy-Bruno O., Schrieffer A.E., Mysorekar I.U. Microbial communities in placentas from term normal pregnancy exhibit spatially variable profiles. Sci Rep, 2017, vol. 7, p. 11200. DOI: 10.1038/s41598-017-11514-4
29. Leiby J.S., McCormick K., Sherrill-Mix S., Clarke E.L., Kessler L.R., Taylor L.J. et al. Lack of detection of a human placenta microbiome in samples from preterm and term deliveries. Microbiome, 2018, vol. 6, p. 196. DOI: 10.1186/s40168-018-0575-4
30. Vanterpool S.F., Been J.V., Houben M.L., Nikkels P.G., De Krijger R.R., Zimmermann L.J., Kramer B.W., Progulske-Fox A., Reyes L. Porphyromonas gingivalis within Placental Villous Mesenchyme and Umbilical Cord Stroma Is Associated with Adverse Pregnancy Outcome. PLoS One, 2016, vol. 11 (1), p. e0146157.
31. Younes J.A., Lievens E., Hummelen R., van der Westen R., Reid G., Petrova M.I. Women and their microbes: the unexpected friendship. Trends Microbiol, 2017, vol. 26, pp. 16–32. DOI: 10.1016/j.tim.2017.07.008
32. Charles M. Cobb, Patricia J. Kelly, Karen B. Williams, Shilpa Babbar, Mubashir Angolkar, Richard J. Derman. The oral microbiome and adverse pregnancy outcomes. International Journal of Women’s Health, 2017, vol. 9, pp. 551–559. DOI: 10.2147/IJWH.S142730
33. Pelzer E., Gomez-Arango L.F., Barrett H.L., Nitert M.D. Maternal health and the placental microbiome. Placenta, 2017, Jun, vol. 54, pp. 30–37. DOI.org/10.1016/j.placenta.2016.12.003