In -vitro Biofilm Formation of Vaginal Isolates of Streptococcus agalactiae; Effect of pH and Culture Media

ABSTRACT

Abstract Background: Streptococcus agalactiae (GBS), a major cause of neonatal morbidity and mortality, is transmitted from mother to neonate via placenta or during birth. Biofilm formation is an important factor in GBS pathogenesis. This study aimed to determine effects of pH, different culture media and nutritional composition on in vitro biofilm forming ability of GBS isolated from pregnant women. Methods: A total of 30 confirmed isolates of GBS from pregnant women were tested for biofilm formation in Todd Hewitt Broth (THB) at pH 4.5,6 and 7. Ten of these isolates were tested for biofilm formation in growth media THB, brain heart infusion broth, tryptic soy broth, Mueller Hinton broth and nutrient broth. Further they were tested for influence of glucose on biofilm formation using crystal violet and MTT assay. Results: Of 30 GBS isolates strong biofilm formation (SBF) was observed at pH 7 in 56.6 %(n=17) while 36.6%(n=11) isolates showed weak biofilm formation (WBF). At pH 4.5, 43.3% (n=13) were non biofilm formers. In THB without glucose, all 10 isolates were SBF while THB with 1% glucose, 3(30%) isolates were SBF, 5(50%) isolates were moderate biofilm producers and 2(20%) isolates were WBF. Ten isolates tested in 5 types of growth media did not show statistically significant difference in biofilm forming ability. Conclusion: All tested vaginal GBS isolates were able to produce biofilms, maximum biofilm formation of GBS was at pH 7.0. and pH 4.5 is not favorable, thus in normal vaginal pH (3.5 - 4.5), GBS finds it difficult to grow biofilms.