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Bacterial colonization of amniotic fluid from intact fetal membranes

  • Joseph M. Miller Jr.
    Affiliations
    Duke University Medical Center, Department of Obstetrics and Gynecology, Division of Perinatal Medicine Durham, North Carolina, USA

    Duke University Medical Center, the Department of Microbiology and Immunology Durham, North Carolina, USA
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  • Marcos J. Pupkin
    Affiliations
    Duke University Medical Center, Department of Obstetrics and Gynecology, Division of Perinatal Medicine Durham, North Carolina, USA

    Duke University Medical Center, the Department of Microbiology and Immunology Durham, North Carolina, USA
    Search for articles by this author
  • Galf B. Hill
    Correspondence
    Reprint requests: Gale B. Hill, Ph.D., P.O. Box 3172, Duke University Medical Center, Durham, North Carolina 27710
    Affiliations
    Duke University Medical Center, Department of Obstetrics and Gynecology, Division of Perinatal Medicine Durham, North Carolina, USA

    Duke University Medical Center, the Department of Microbiology and Immunology Durham, North Carolina, USA
    Search for articles by this author
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      Abstract

      Except in rare instances, intact membranes have been considered to be a barrier to infection of amniotic fluid. Amniotic fluid was collected from 45 selected patients by amniocentesis or needle amniotomy prior to or duing labor, or by needle aspiration at the time of cesarean section. Fluid was cultured and examined directly by Gram stain. Among 14 patients who were not in labor, only one had growth of bacteria from broth culture [<10 colony-forming units per milliliter (CFU/ml) of amniotic fluid]. Among amniotic fluid cultures from 31 patients in labor, 13 were positive on primary plating media (>102 CFU/ml), five grew in broth only, and 13 were negative. Of the 13 patients with positive cultures of >102 CFU/ml, eight patients had clinical chorioamnionitis; five patients were afebrile and clinically asymptomatic, except that premature labor occurred in three of these patients. Of the amniotic fluid cultures with >102 CFU/ml, six contained only aerobes, five contained only anaerobes, and two contained mixed aerotolerance types. Aerobic bacteria included Haemophilus influenzae, Pseudomonas aeruginosa, Klebsiella pneumoniae, Listeria monocytogenes, group B β-hemolytic streptococci, and coagulase-negative staphylococci. Anaerobic organisms isolated included Fusobacterium nucleatum (on three occasions), Bacteroides corrodens, Bacteroides ochraceus, Bacteroides bivius, and Peptostreptococcus micros. Bacteria and leukocytes observed by oil immersion microscopy on direct Gram stain of unspun amniotic fluid were significantly associated with culture of >102 CFU/ml. Also, the presence of bacteria on Gram stain and >102 CFU/ml from culture were significantly associated with the presence of clinical chorioamnionitis. These data demonstrated a wider spectrum of bacteria capable of colonizing amniotic fluid in the presence of intact membranes than was previously appreciated, indicating that direct Gram stain in addition to culture can provide valuable diagnostic information.
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