[Isolation and characterization of vaginal lactobacilli producing hydrogen peroxide].
OBJECTIVE
Isolation and characteristics of vaginal lactobacilli that actively generate H2O2 and have high antagonistic activity.


METHODS
Staphylococcus aureus 8956, Escherichia coli 8852, Klebsiella pneumoniae 8795 and Candida albicans 5646 were used as target-strains. Skim milk and MRS medium were used for lactobacilli isolation and cultivation. Antagonism was studied in complete agar and Saburo medium. Merckoquant peroxide test (Merck) stripes were used for the determination of H2O2. Antibacterial activity was determined by diffusion into agar. Specific culture growth rate was determined by conventional method, acidification of the culture medium--by pH-meter.


RESULTS
12 strains were isolated from vaginal smears of healthy women. These strains have an ability to ferment milk among which a highly active H2O2 producer was isolated and attributed to Lactobacillus delbrueckii by the results of 16S rRNA and alpha-subunit RNA polymerase gene sequence analysis (16S rDNA and rpoA genes are registered in GenBank, numbers HQ379171 and HQ379180 respectively). L. delbrueckii MH-10 bacterial cells were characterized by specific growth speed 1.26 per hour, reaching a maximum titer of 2 x 10(9) PFU/ml with lowering medium pH to 4.0. Under aerated conditions H2O2 concentration reached 100 microg/ml or more. L. delbrueckii MH-10 has high antibacterial activity against S. aureus, E. coli, K. pneumoniae.


CONCLUSIONS
L. delbrueckii MH-10 isolate is an active H2O2 producer, has high growth speed and broad antibacterial activity spectrum, is a perspective candidate for the development of probiotic preparation for the prophylaxis and therapy of vaginoses.
M M
MM
Pashaian
G G
GG
Oganesian
D000818:Animals; D000898:Antibiosis; D015373:Bacterial Typing Techniques; D002176:Candida albicans / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects* / Q000254:growth & development; D005260:Female; D005285:Fermentation; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D007711:Klebsiella pneumoniae / Q000187:drug effects*; D048668:Lactobacillus delbrueckii / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008892:Milk / Q000378:metabolism; D019936:Probiotics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D013211:Staphylococcus aureus / Q000187:drug effects*; D014621:Vagina / Q000382:microbiology*; D016585:Vaginosis, Bacterial / Q000382:microbiology* / Q000628:therapy
D012336:RNA, Ribosomal, 16S; D006861:Hydrogen Peroxide