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Copyright © 2022 Liu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. 20220523 2022 ABSTRACT Recurrent spontaneous abortion (RSA) is a complex multifactorial disease. Recently, the microbiota of the female reproductive tract, as an emerging factor in RSA, has gradually attracted the attention of many clinical researchers. Here, we reported that the microbiota of the lower and upper female reproductive tracts from patients with RSA showed no significant differences in alpha diversity compared to that of controls. Beta diversity was significantly higher in the RSA group than in the control group in the vaginal microbiota (P = 0.036), cervical microbiota (P = 0.010) and microbiota from uterine lavage fluid (P = 0.001). In addition, dramatic decreases in gamma interferon and interleukin-6 cytokine levels were observed in the RSA group. In conclusion, our data suggested altered microbial biodiversity in the vagina, cervix and uterine lavage fluid in the RSA group. Alterations in the microbiota in the uterine cavity could be associated with altered cytokine levels, which might be a risk factor for RSA pathogenesis. Moreover, the microbiota composition differed markedly from the lower genital tract to the uterine cavity, and the microbiota in the uterine cavity also distinctly varied between endometrial tissue and uterine lavage fluid in the RSA group. Hence, sampling with these two methods simultaneously allowed a more comprehensive perspective of microbial colonization in the uterine cavity. IMPORTANCE As an obstacle to pregnancy, recurrent spontaneous abortion (RSA) can be caused by a variety of factors, and a current understanding of the etiology of RSA is still lacking; half of cases have an unknown cause. A substantial fraction of patients show no improvement after treatment. Since the microbiota of the female reproductive tract has been proposed as an emerging factor in RSA patients, further investigation is needed to provide guidance for clinical therapy. In general, this is the first report describing the distinct alterations of the vaginal, cervical, and uterine microbiota in RSA, not just that in the vagina. Furthermore, another major strength of this study derived from the further in-depth investigation and analysis of the characteristics of the microbiota colonizing the upper female genital tract in RSA, which provided a more comprehensive view for investigating the uterine microbiota. 20220205 20220328 2165-0497 Adobe LiveCycle PDF Generator; modified using iText® 5.5.13.2 ©2000-2020 iText Group NV (AGPL-version)2022-08-26T00:12:56-07:00 endstream endobj 19 0 obj <>stream x+| endstream endobj 20 0 obj <>stream xS**T0T0Biy, endstream endobj 21 0 obj <>stream x+| endstream endobj 22 0 obj <>stream xS**T0T0BihuU endstream endobj 23 0 obj <>stream x+| endstream endobj 24 0 obj <>stream xS**T0T0Biy8# endstream endobj 25 0 obj <>stream x+| endstream endobj 26 0 obj <>stream xS**T0T0Bih~V endstream endobj 27 0 obj <>stream x+| endstream endobj 28 0 obj <>stream xS**T0T0BiyJ% endstream endobj 29 0 obj <>stream x+| endstream endobj 30 0 obj <>stream xS**T0T0Biy+ endstream endobj 31 0 obj <>stream x+| endstream endobj 32 0 obj <>stream xS**T0T0Biyw* endstream endobj 33 0 obj <>stream x+| endstream endobj 34 0 obj <>stream xS**T0T0BihX endstream endobj 35 0 obj <>stream x+| endstream endobj 36 0 obj <>stream xS**T0T0Biye( endstream endobj 37 0 obj <>stream x+| endstream endobj 38 0 obj <>stream xS**T0T0BihlT endstream endobj 39 0 obj <>stream x+| endstream endobj 40 0 obj <>stream xS**T0T0BiyS& endstream endobj 41 0 obj <>stream x+| endstream endobj 42 0 obj <>stream xS**T0T0Biyn) endstream endobj 43 0 obj <>stream x+| endstream endobj 44 0 obj <>stream xS**T0T0Biy\' endstream endobj 45 0 obj <>stream x+| endstream endobj 46 0 obj <>stream xS**T0T0BiyA$ endstream endobj 47 0 obj <>stream x+| endstream endobj 48 0 obj <>stream xS**T0T0BihW endstream endobj 50 0 obj <>stream HWێE79̾8ƁwdN(%u /~~oNU5% R9UoQmNÛ?޼`7j;)61q\MmfkMm˛cY]?=8ȷU=|{{EɵnIئ4޷NQvMY{r;4I":CȁN+w-}fc𫫻+[߄;1ǣX6׵HQ]VO$[|2(*sRIح*^H lS۽" 71&ѻzW 5bwpo ήwǮw Eܳ*bi\P:*D8C^(' @dSF?wW=FUO@"&T\nS#d:dz<[sj <\ʶb9G(6qp\[  -sng]O}S9MW}C3aR]Am^$:1fڏd^K uqO 2[ASMCLӂkNY>iƠ3տ^Q Uit:!s܇F8\hy>F|LLXatv%m:\PxJc_jAeѥ<6Yo T e~HW}"ۗ`|vfMԀJt!z ̢odEDNBjR!!/p$UrUƟ#sW-q%*/)y6@/^$-<-N(7%O-lyMiD2&jFkLkJ,NpcWqa_ʙ5ǟ>

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