The group B Streptococcus is one of the most virulent organisms causing perinatal infection. Human amniotic fluid from the second and third trimesters was pooled and analyzed for electrolytes, protein, albumin, zinc, inorganic phosphorus, ferritin, lysozyme, and immunoglobulins. We inoculated replicates of specimens with known virulent strains of group B streptococci (893, 891, and 878) and Escherichia coli (C5) with Todd-Hewitt broth and normal saline solution used as controls. Group B streptococci strains 893 and 891 proliferated rapidly at rates similar to their rates in Todd-Hewitt Broth. Strain 878 grew at a rate slower than that of strains 893 and 891. The amniotic fluid specimens were similar with respect to factors reported as inhibitory to bacterial proliferation. Second- and third-trimester amniotic fluid supports the growth of group B streptococci as well as a culture medium optimized for bacterial growth. Strain-specific variance in group B streptococci growth rates in amniotic fluid may have clinical significance for those at risk for group B streptococci infection.