Outline Vaginal ecosystem Estrogen and recurrent UTI in women What are probiotics and how do they work? Current proposed uses and a look at some of the evidence Issues in prescribing their use
Health vaginal ecosystem Dynamic Equilibrium exists between: Epithelium Normal colonizing organisms (mostly Lactobacilli SPP.) Local secretory and celluar immune factors Vaginal pH maintained acidic (3.8 ~ 4.2) creates unfavourable environment for pathogen
Normal Vaginal flora LACTOBACILLI Gram positive rods Excrete hydrogen peroxide Present in 100% of women with normal flora Affects adherence of epithelial cells Protect against bacterial/candidal infections
Facultative organisms Diphtheriods Coagolase negative Staphycococci Streptococci E. coli Ureaplasma urealyticum Mycoplasma hominis
Anaerobic organisms Present in low, non-pathogenic concentrations Peptostreptococus Bacteroides Fusobaterium Mobiluncus Gardnerella (40% - 60%) in normal secretions
UTI in women• 1 billion each year (Reid, 2003)• > 300 million cases annually worldwide (Reid 2001. Am J Clin Nutr 73: S437-S443)• Each episode: on average 6 days of symptoms• Sequelae: pyelonephritis --> preterm birth• Uropathogens: E. coli (approx. 70%), Enterobacteriacae Enterococcus faecalis, Staphylococcus spp.• Increasing drug resistance among uropathogens
Risk factors for BV The vagina is not a steady state ecosystem. Menstruation cyclic changes in the vaginal environment Levels of estrogen and progesterone alter, Changing the environment for lactobacilli by influencing cell surface receptor expression levels of glyocgen and glucose as substrate levels of vaginal pH Estrogen protective: BV prevalence lower in women using oral contraception (Yen et al. 2003) BV lower during pregnancy (3rd trimester) (Hay et al. 1994) Clinical trials with estradiol cure of BV, restoration pH, ... (Kanne & Jenny 1991, Parent et al. 1996, Ozkinay et al. 2005)
Risk factors for BV Race/ethnicity (black women at higher risk): In Africa: > 50% of women have BV (Paxton et al. 1998) L. iners instead of L. crispatus? (Anukam et al. 2005) genetic differences in epithelial surface molecules --> different adherence? Frequency and kind of intercourse: new male sexual partner more male sexual partners female sexual partner * Sexual intercourse once a week was the only risk factor associated with loss of H2O2 producing lactobacilli. (Vallor et al. 2001) --> BV is STD? or raise of pH?
Sequelae of BV (and UTI)↑ HSV2 infection (Cherpes TL 2005. CID 40: 1422)↑ HIV shedding (Cu-Uvin S. 2004. CID 33: 894, Sewankambo. 1997. Lancet 350: 546) --> ↑ Perinatal mother-child HIV-transmission --> ↑ Sexual HIV-transmission↑ Susceptibility for hiv-infection↑ Infection with CT and HPV (da silva CS. 2004. Goinvest. 58: 189) ↑ Early loss after IVF (Eckert LO. 2003. IDOG 11: 11-17)Associated with recurrent UTI (Hooton TM. 2001. IJAA 17: 259-268)Associated with PID, postpartum endometritis ...
Estrogens: Clinical trials A controlled trial of intravaginal estriol in postmenopausalwomen with recurrent UTIs Raz R, Stamm WE. N Engl J Med 1993, 329:753-756 •93 postmenopausal women with a history of recurrent UTIs •Randomized, double-blind, placebo-controlled trial of topical intravaginal estriol cream UTI incidence; 0.5 vs. 5.9 episodes per patient-year Lactobacilli restoration after one month; 61% vs 0% Vaginal pH declined (from 5.5 to 3.8) Vaginal Enterobacteriaceae fell from 67 to 31% with estriol but unchanged (from 67 to 63 %) with placebo •prevents recurrent UTI in postmenopausal women, probably by modifying the vaginal flora.
Probiotics: definitions World Health Organization: “live microorganisms which when administered in adequate amounts confer a health benefit on the host” A bacterial strain that: survives the stomach acid and bile adheres to intestinal lining grows and establishes temporary residence in the intestines imparts health benefitsR Fuller. Probiotics: The Scientific Basis. London: Chapman and Halls. 1992
Probiotics: History Pasteur (1877) antagonistic interaction between bacterial strains non-pathogenic bacteria should be used to control pathogenic bacteria Metchnikoff (1907) lactic fermentation of milk arrested putrefaction consumption of fermented products would offer the same benefit to humans longevity in Bulgarian peasants was due to ingestion of “soured milks” 1980’s Fuller establishes first definition of probiotics.
Probiotics: Potential MOA Inhibition of adhesion Immunomodulation Production of antimicrobial substances Modification of toxins or toxin receptors Competition for nutrients Reduction in bacterial translocation Anti-inflammatory signaling within the epithelium
Probiotics: Prescribing Which organism to use? Which product? For what conditions? What dose? How long? Any side effects to be aware of? Cost of therapy?
Vaginal probiotics: Clinical trials Treatment of BV with lactobacilli Hallen A, et al. Sex Transm Dis. 1992, 19:146-148. • 60 women with BV double blind, placebo-controlled trial • Immediately after completion of treatment, 16/28 women treated with lactobacilli had normal vaginal wet smear results none of the 29 women treated with placebo. • Only 3 of the women who received the Lactobacillus suppository were free of BV after the subsequent menstruation.
Vaginal probiotics: Clinical trials Vaccination against nonspecific BV; Double-blind study ofGynatren Siboulet A. Gynakol Rundsch. 1991;31(3):153-160 • 167 patients with nonspecific BV vaccinated with Gynatren, a Lactobacillus vaccine double-blind, randomized, placebo- controlled trial • During the study period of 14 months, vaccination was significantly better than the placebo with regard to its therapeutical effect. • Vaccination with Gynatren is effective to prevent recurrences of nonspecific vaginosis .
Vaginal probiotics: Clinical trials An audit of Gynatren (a L. acidophilus lyophilisate) vaccination inwomen with recurrent bacterial vaginosis. Pattman RS et al. Int J STD AIDS. 1994, 5: 299 Bacterial vaginitis: protection against infection and secretoryimmunoglobulin levels in the vagina after immunization therapy withGynatren. Ruttgers H. Gynecol Obstet Invest. 1988, 26: 240-9 Effect of Lactobacillus immunotherapy on genital infections inwomen (Solco Trichovac/Gynatren ). Karkut G. Geburtshilfe Frauenheilkd. 1984, 44: 311-4 Randomized double-blind study on the prevention of reinfection intrichomoniasis using Solco Trichovac vaccination. Litschgi M. Gynakol Rundsch. 1982, 22: 70-3
Vaginal synbiotics: Clinical trials Therapy of bacterial vaginosis (BV) using exogenously- applied L. acidophili and a low dose of estriol : aplacebo-controlled multicentric clinical trial. Parent D, et al. Arzneimittelforschung. 1996, 46: 68-73 •Vaginal tablets (Gynoflor) containing 50 mg of a lyophilisate of viable, H2O2-producing L. acidophilus (>107 CFU/tablet) and 0.03 mg estriol •32 non-menopausal women with BV •6-day therapy with 1~2 vaginal tablets daily cure rate 2 wks after the start of therapy ; 77% vs 25% cure rate 4 wks after the start of therapy ; 88% vs. 22% •A significant increase in the number of lactobacilli
Vaginal synbiotics: Clinical trials The effectiveness of live lactobacilli in combination with low dose estriol (Gynoflor) to restore the vaginal flora after treatment of vaginal infections. Ozkinay E, et al. Brit. J. Obstetr. Gynaecol. 2005, 112: 234-40 •Randomised, placebo-controlled, double-blind clinical trial •360 women with vaginal infections •Restoration therapy with live lactobacilli in combination with low dose estriol (n = 240) or placebo (n = 120) 2~7 days after the end of the anti-infective therapy •Follow up at 3~7 days and 4~6 weeks after the end of the restoration therapy •Restoration of the vaginal flora can be significantly enhanced by the administration of live lactobacilli in combination with low dose estriol.
Non-Vaginal probiotics: Clinical trials with Yoghurt Ecological treatment of bacterial vaginosis. Chimura T, et al. Jpn J Antibiot. 1995, Mar;48: 432-6 • 11 women with BV: intravaginal application of 5 ml of commercial yoghurt (pH 4.3). • Evaluation after 3 days: vaginal discharge and bacteriology Significant decrease of vaginal discharge and redness Vaginal pH was lowered significantly. Overall bacteriological effects: 6 (54.5%) were eradicated, 3 were partly eradicated, 2 were replaced. • Lactobacillus therapy was effective in both clinical and bacteriological responses.
Non-Vaginal probiotics: Clinical trials with Yoghurt Bacterial vaginosis in pregnancy treated with yoghurt. Neri A, et al. Acta Obstet Gyecol Scand 1993, 72: 17-22 •32 women with BV in the first trimester of pregnancy were treated with intravaginal application of yoghurt . •The result was favorable. Ingestion of yogurt containing Lactobacillus acidophilus as prophylaxis for candidal vaginitis. Hilton E, et al. Ann Intern Med 1992, 116: 353-7 •Daily ingestion of 8 ounces of yogurt containing L. acidophilus decreased both candidal colonization and infection.
Non-Vaginal probiotics: Clinical trials Augmentation of antimicrobial MDZ therapy of BV with oralprobiotic L. rhamnosus GR-1 and L. reuteri RC-14:randomized, double-blind, placebo controlled trial. Anukam K, et al. Microbes Infect. 2006 •125 premenopausal women with BV •Oral MDZ (500 mg) bid for 7 d + oral L. rhamnosus GR-1 (1x109) and L. reuteri RC-14 (1x109) or placebo bid for 30 d Cure rate of BV; 88% vs 40% (p < 0.001). Recovery of Lactobacillus sp. counts (> 10 5 cfu/ml) 96% vs 53% •Use of lactobacilli and antibiotic in the eradication of BV in black African women is efficacious .
Non-Vaginal probiotics: Clinical trials Oral use of L . rhamnosus GR-1 and L. fermentum RC-14 significantly alters vaginal flora: randomized, placebo-controlled trial in 64 healthy women. Reid G, et al. FEMS Immunol Med Microbiol. 2003, 35: 131-4 64 healthy women Oral capsules of L. rhamnosus GR-1 and L. fermentum RC-14 given daily for 60 days Restoration from asymptomatic BV microflora to a normal lactobacilli colonized microflora; 37% vs 13% (P=0.02). A significant increase in vaginal lactobacilli at day 28 and 60 A significant depletion in yeast at day 28 A significant reduction in coliforms at day 28, 60 and 90 for lactobacilli-treated subjects
Non-Vaginal probiotics: Clinical trials In vitro testing of Lactobacillus acidophilus NCFM as a possibleprobiotic for the urogenital probiotic applications. Reid G. Int Dairy J. 2000, 10: 415-9 Oral probiotics can resolve UTI. Reid G. FEMS Immunol 2001, 30:49-52 Assessment of the capacity of lactobacilli to inhibit growth ofuropathogens. Osset J et al. JID 2001, 183: 485-91 Probiotic feeding reduces UTI in preterm infants. Dani C et al. Biol Neonate 2002, 82: 103-8 Probiotics: alternative treatment in urology. Hoesl & Altwein. Eur Urol 2005, 47: 288-96
Vaginal probiotic products CTV-05: L. crispatus 108 cfu/capsule (Chrisope Tech., La & Gynelogix, Colo.) Fem-Dophilus: L. rhamnosus GR-1™ and L. reuteri RC-14™ (Urex Biotech) Florajen: L. acidophilus 2 x 1010/capsule (American Lifeline, Wisconsin) Gy-Na-Tren: L. acidophilus 2 x 109 cfu/capsule (Vitalis, Nederland) Gynoflor: L. acidophilus 107 cfu + 0.03 mg estriol/tablet (Mithra, Liège, from Grünenthal,produced by Medinova (Zürich, S) Infemin: L. crispatus, L. acidophilus, L. fermentum, L. rhamnosus , all at 109 cfu/ml(Pierre Fabre Sante, Boulogne, France) Intrafresh: vaginal probiotic pessarium: PEG + L. acidophilus (BioCare, UK) Lacto AC: 10 species/capsule. TJCP: The Jordan Prentice Co., Fl. LactoFem: L. Rhamosus, L. acidophilus both at 109/capsule (Mithra, Belgium) Trenev Trio (Vitals, Nederland)
Vaginal probiotics: Rationale todevelop Clear hypothesis about role of lactobacilli Clearly established protective role of lactobacilli Lactobacilli are predominant in the vagina. Application can be topical reaching high inocula no organoleptic considerations Easy to perform clinical trials: Easy sampling Easy re-isolation of probiotic lactobacilli after application Already available
Probiotics vs antibiotics Antibiotics damage commensal microflora. can increase the occurrence of resistant bacteria can have adverse side effects Probiotics can be used in adjunction to antibiotics to restore the commensal microflora
Probiotics: Practical Issues Not FDA regulated Quality control is poor 80% of preparations tested had 1% or less of the bacterial concentration on the label Numerous preparations on the market Which strains work best? Do different strains work better for different diseases? Do combinations work better than single strains? May need several months of therapy to see an effect Likely stop working after discontinued Concentration (dose) highly variable
Conclusions The use of probiotics in prevention of UTI is promising. Should be used carefully and cautiously, and only on the basis of strong scientific evidence. Response is species specific; the success of one species of Lactobacillus in a certain application does not imply that all related strains of this species will be capable of producing a comparable response. Carefully conducted double-blind, placebo-controlled studies to document the individual efficacy of each specific organism for each potential clinical application are needed.