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Semeshchenko D.,Catedra de Microbiologia y Parasitologia | Santalucia M.,Catedra de Microbiologia y Parasitologia | Marques M.,Catedra de Microbiologia y Parasitologia | Farinati A.,Catedra de Microbiologia y Parasitologia | Orsini A.,Catedra de Microbiologia y Parasitologia
Prensa Medica Argentina | Year: 2014

The biofilm (BF) lifestyle of microbial flora in vaginal tract plays an important role in the reproductive health of sexually active women (SAW). We previously study the BF in the genital tract in different situations mainly under antimicrobial stress. Our objective is to describe the vaginal BF (VBF) 1-from starting in SAW without genital infections and 2-the behavior of VBF in normal cases and in endogenous infections under challenged by estradiol hemisuccinate (EH) and fluconazole (FCZ). Methods: vaginal samples were collected from 37 SAW: normal flora (NF) 16, endogenous infections 21: candidiasis (WC) 10; bacterial vaginosis (BV) 11. The VBF was investigated using vaginal layers in glass coupon (GC) for each patient (previously described) and microtiter assays (MA) for quantitative studies. The glass coupon (GC) was immediately placed in Trypticase Soy Broth pH 6.5, in Man Rogosa Sharpe pH 4.5 and in Sabouraud Broth. We use EH (200 μg/ml) and FCZ (6 ug/ml) as stress agents. The last was employed in WC cases using Candida albicans (CA) and non albicans Candida (NAC) isolates as control with similar techniques. Escherichia coli (EC) as common microorganism from genital tract were used to form mixed biofilm with CA and NAC. Everything was read by optical microscopy previous tinction with crystal violet and Gram after 24 and 48 hours. Besides in WC cases and their controls, GC were stained with Gram, acridine orange (AO) and use fluorescence microscope. Results: Gram positive cocci (GPC) initiate the BF formation in NF, WC and BV. Activity of EH: In NF interferes with GPC, and gram positive bacilli in the starting of BF. In BV we detected BF pieces detachment and dispersion. In WC: EH estimulation at both pH was observed (BF with blastospores, pseudohyphae and hyphae). BF formation in all Candida isolates was detected but with different constitution. The exopolisacharide (EPS) was better demonstrated with AO and was more evident in NAC without influence of EH. EH improves EC BF in the early stage but at 24 hours fragmentation and dispersion was shown Activity of FCZ: we observed that EC form mixed BF with CA and NAC but was more evident with NAC. FCZ has not influence on adherence or dispersion in mixed BF but has reduced activity in CA and NAC BF monospecies. Conclusions: Despite Lactobacillus are predominant bacterial species in the vagina, GPC's are the first to appear in BF. Our observations suggest that EH can function as promoter or inhibitor of BF in according to developmental process and pathologies. The BVBF dispersion and detachment as the dispersion of EC BF are factors to considerer for therapeutic proposals. CA BF is always promoted by EH. These data suggest that EH could induce alteration in fungal receptor sites. FCZ has not influence in mixed BF and we think that FCZ activity can be reduced by BF presence and not by EC modulation.

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