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Modeling Ascending Vaginal Infection, Preterm Birth, and Neonatal Morbidity in Mice
Published on: October 10, 2025
Evidence that intra-amniotic infections are often the result of an ascending invasion - a molecular microbiological
Roberto Romero1,2,3,4,5,6, Nardhy Gomez-Lopez1,7,8,9, Andrew D Winters1,8
1Perinatology Research Branch, Division of Obstetrics and Maternal-Fetal Medicine, Division of Intramural Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, U.S. Department of Health and Human Services, Bethesda, MD and Detroit, MI, USA.
Abstract:
Background Microbial invasion of the amniotic cavity resulting in intra-amniotic infection is associated with obstetrical complications such as preterm labor with intact or ruptured membranes, cervical insufficiency, as well as clinical and histological chorioamnionitis. The most widely accepted pathway for intra-amniotic infection is the ascension of microorganisms from the lower genital tract. However, hematogenous dissemination of microorganisms from the oral cavity or intestine, retrograde seeding from the peritoneal cavity through the fallopian tubes, and introduction through invasive medical procedures have also been suggested as potential pathways for intra-amniotic infection. The primary reason that an ascending pathway is viewed as most common is that the microorganisms most often detected in the amniotic fluid are those that are typical inhabitants of the vagina. However, thus far, no studies have shown that microorganisms in the amniotic cavity are simultaneously present in the vagina of the woman from which they were isolated. The objective of the study was to determine the frequency with which microorganisms isolated from women with intra-amniotic infection are also present in the lower genital tract. Methods This was a cross-sectional study of women with intra-amniotic infection with intact membranes. Intra-amniotic infection was defined as a positive culture and elevated concentrations of interleukin-6 (IL-6) (>2.6 ng/mL) in amniotic fluid and/or acute histologic chorioamnionitis and funisitis. Microorganisms isolated from bacterial cultures of amniotic fluid were taxonomically identified through matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) and 16S ribosomal RNA (rRNA) gene sequencing. Vaginal swabs were obtained at the time of amniocentesis for the identification of microorganisms in the lower genital tract. The overall bacterial profiles of amniotic fluids and vaginal swabs were characterized through 16S rRNA gene sequencing. The bacterial profiles of vaginal swabs were interrogated for the presence of bacteria cultured from amniotic fluid and for the presence of prominent (>1% average relative abundance) operational taxonomic units (OTUs) within the overall 16S rRNA gene bacterial profiles of amniotic fluid. Results (1) A total of 75% (6/8) of women had bacteria cultured from their amniotic fluid that are typical residents of the vaginal ecosystem. (2) A total of 62.5% (5/8) of women with bacteria cultured from their amniotic fluid also had these bacteria present in their vagina. (3) The microorganisms cultured from amniotic fluid and also detected in the vagina were Ureaplasma urealyticum, Escherichia coli, and Streptococcus agalactiae. (4) 16S rRNA gene sequencing revealed that the amniotic fluid of women with intra-amniotic infection had bacterial profiles dominated by Sneathia, Ureaplasma, Prevotella, Lactobacillus, Escherichia, Gardnerella, Peptostreptococcus, Peptoniphilus, and Streptococcus, many of which had not been cultured from the amniotic fluid samples. (5) Seventy percent (7/10) of the prominent (>1% average relative abundance) OTUs found in amniotic fluid were also prominent in the vagina. Conclusion The majority of women with intra-amniotic infection had bacteria cultured from their amniotic fluid that were typical vaginal commensals, and these bacteria were detected within the vagina at the time of amniocentesis. Molecular microbiological interrogation of amniotic fluid from women with intra-amniotic infection revealed that the bacterial profiles of amniotic fluid were largely consistent with those of the vagina. These findings indicate that ascension from the lower genital tract is the primary pathway for intra-amniotic infection.
Insights
Intra-amniotic infection often involves bacteria from the lower genital tract, with 62.5% of cases showing the same bacteria in both the amniotic fluid and the vagina. This confirms the ascending pathway as the primary route for infection.
Area of Science:
- Obstetrics and Gynecology
- Infectious Diseases
- Microbiology
Background:
- Intra-amniotic infection is linked to preterm labor and chorioamnionitis.
- The ascending pathway from the lower genital tract is considered the most common route for intra-amniotic infection.
- Previous studies lacked direct evidence of simultaneous microbial presence in amniotic fluid and the vagina.
Purpose of the Study:
- To determine the frequency of microorganisms in amniotic fluid also being present in the lower genital tract of women with intra-amniotic infection.
- To investigate the microbial profiles of amniotic fluid and vaginal swabs using advanced molecular techniques.
Main Methods:
- Cross-sectional study of women with intra-amniotic infection and intact membranes.
- Amniotic fluid and vaginal swabs were collected for bacterial culture and 16S ribosomal RNA (rRNA) gene sequencing.
- Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) was used for taxonomic identification.
Main Results:
- 75% of women had bacteria in their amniotic fluid that are common vaginal residents.
- 62.5% of women showed the same cultured bacteria in both amniotic fluid and their vagina (Ureaplasma urealyticum, Escherichia coli, Streptococcus agalactiae).
- 16S rRNA gene sequencing revealed consistent bacterial profiles between amniotic fluid and vaginal swabs, with 70% of prominent operational taxonomic units (OTUs) overlapping.
Conclusions:
- The majority of intra-amniotic infections involve bacteria originating from the lower genital tract.
- The findings strongly support the ascending pathway as the primary route for intra-amniotic infection.
- Molecular analysis confirms significant overlap in microbial communities between amniotic fluid and the vagina in infected women.
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