Kyoko Kawazoe

Studies on the clinical implications of anaerobes, especially Prevotella bivia, in obstetrics and gynecology

Prevotella bivia is one of the anaerobic bacteria found in the resident flora of the female genital tract. The detection rate ofP. bivia has been poor, however, mainly due to problems such as the poor survival of this microbe in the presence of oxygen. To identifyP. bivia, it is necessary to start the culture, within 1 hour after sampling. We were able to isolate 13 strains ofP. bivia (32.5%) from 40 cases of preterm premature rupture of membrane, 8 strains (40.0%) from 20 cases of puerperal uterine endometritis, 20 strains (40.0%) from 50 cases of pyometra, 6 strains (40.0%) from 15 cases of severe pelvic inflammatory disease, and 6 strains (17.1%) from 35 cases of bacterial vaginosis. Most clinical isolates ofP. bivia were susceptible to the commonly used antimicrobial agents.P. bivia was detected concomitantly with aerobic bacteria, related to the alteration of vaginal microbial flora during the menstrual cycle.P. bivia increased in the follicular phase of the menstrual cycle, suggesting that the growth of this microbe might be partly regulated by sex steroids.

Elastase activity of anaerobes isolated from amniotic fluid with preterm premature rupture of membranes

OBJECTIVE A total of 131 anaerobes isolated from amniotic fluid with preterm premature rupture of membranes and stored were examined for elastolytic activity by the method described by Williams et al (Lett Appl Microbiol 1988;7:173-6). STUDY DESIGN Each strain was spot inoculated on a Columbia blood agar plate containing 1% solubilized elastin and incubated for 5 days under anaerobic conditions. Undigested elastin was precipitated by flooding trichloroacetic acid solution onto the plate, and a clear zone was visible as the elastolytic reaction around the spot of bacterial growth. RESULTS Ninety-three (71.0%) of 131 organisms showed a positive elastolytic reaction. Eleven of 20 strains (55.0%) of Peptostreptococcus magnus, 9 of 18 strains (50.0%) of Peptostreptococcus micros, 12 of 12 strains (100.0%) of Fusobacterium nucleatum, 15 of 28 strains (53.6%) of Bacteroides fragilis, 8 of 15 strains (53.3%) of Bacteroides thetaiotaomicron, and 38 of 38 strains (100.0%) of Prevotella bivia were elastolytic. CONCLUSION Anaerobic bacterial species prevalent in the normal vaginal flora that were isolated from amniotic fluid of women with preterm rupture of membranes produced elastolytic activity, plausibly inducing the destruction of host constitutive components.

Preterm labor and bacterial intraamniotic infection: Arachidonic acid liberation by phospholipase A2 of Fusobacterium nucleatum ☆ ☆☆

OBJECTIVE The studies presented in this report were undertaken to evaluate whether Fusobacterium nucleatum, a common anaerobic isolate in intrauterine infection, stimulates arachidonic acid metabolism, a rate-limiting step for prostaglandin synthesis, in the human uterine endometrium. STUDY DESIGN Effects of F nucleatum on arachidonic acid liberation from human uterine endometrial cells and of F nucleatum extract on lysophosphatidylcholine production in human uterine endometrial cells were investigated. RESULTS When human uterine endometrial cells labeled with tritiated arachidonic acid to an isotopically steady state were exposed to an extract of F nucleatum, arachidonic acid liberation was stimulated, accompanied by lysophospholipid formation. Similar stimulatory effects on phospholipid degradation were also observed in the experiment with bacterially conditioned media. CONCLUSIONS These results suggest that F nucleatum stimulates endometrial phospholipid metabolism, related to activity of phospholipase A2, which might induce the onset of labor associated with intraamniotic infection.

Studies on the pathogenicity of anaerobes, especially Prevotella bivia, in a rat pyometra model.

OBJECTIVE: Prevotella bivia is one of the anaerobic bacteria that resides in the flora of the female genital tract. We studied the pathogenicity of P. bivia in a rat pyometra model. METHODS: The experimental animal (rat) model of pyometra was developed to investigate the pathogenicity of P. bivia in a rat pyometra model. RESULTS: In the groups inoculated with aerobes alone, the infection rate was 10% (1/10) in the Staphylococcus aureus- or Staphylococcus agalactiae-inoculated group and 20% (2/10) in the Escherichia coli-inoculated group. Infection was not established in the groups inoculated with anaerobes alone. High infection rates were observed in all the mixed-infection groups. In the S. agalactiae- and Bacteroides fragilis-, S. agalactiae- and P. bivia-, F. coli- and B. fragilis-, and E. coli- and P. bivia-inoculated groups, an infection rate of 100% (10/10) was demonstrated. The efficacy of antibiotics such as flomoxef (FMOX) could be determined using a rat pyometra model. In relation to the alteration of vaginal microbial flora during the menstrual cycle, estrogen increased the growth of P. bivia. CONCLUSION: Mixture of aerobic bacteria and P. bivia increased the pathogenicity of P. bivia. Estrogen would be useful for raising up the inflammatory change of the uterus in experimental models of genital tract infection due to P. bivia.

Comparative Study on the Effectiveness of Antifungal Agents in Different Regimens against Vaginal Candidiasis

Objective: A study was carried out to compare three treatment regimens for vaginal candidiasis. Methods: A total of 150 women with clinical and mycological evidence of vaginal candidiasis were randomized to receive daily a 200-mg dose of oral itraconazole for 3 days (50 women), a single oral 150 mg dose of fluconazole (50 women), or daily 100 mg dose of intravaginal clotrimazole for 6 days (50 women). They were assessed at 5–15 days (short-term assessment) and again at 30–60 days (long-term assessment) after discontinuation of the treatment. Results: At the short-term or long-term assessment, Candida species were completely eradicated from the vagina in 80 or 74% in the 3-day oral itraconazole group, 76 or 70% in the single oral fluconazole group, and 72 or 60% in the intravaginal clotrimazole group, respectively. The rates of clinical effectiveness were 92 or 88% in the 3-day oral itraconazole group, 80 or 76% in the single oral fluconazole group, and 72 or 58% in the intravaginal clotrimazole group, respectively. Treatment-related side effects were not found in any group. Conclusions: Our study suggests that the treatment of vaginal candidiasis with oral itraconazole or oral fluconazole would be effective and that an oral itraconazole or fluconazole therapy might be one choice in the treatment of vaginal candidiasis.

Comparative Study on Vaginal or Oral Treatment of Bacterial Vaginosis

The bacterial epidemiology of bacterial vaginosis (BV) and the efficacy of vaginal or oral treatment of BV with clindamycin (CLDM) were investigated. The epidemiology of BV was investigated in 100 symptomatic women before CLDM therapy. Two groups consisting of 50 patients each with the diagnosis of symptomatic BV were treated with either oral administration of 450 mg CLDM three times daily or 2% CLDM phosphate in vaginal cream (self-made) 5 g once a day, for 7 days. There was no significant differences in efficacy among vaginal and oral therapies with CLDM. Vulvovaginal irritation occurred in 3 patients orally treated and in 1 patient vaginally treated. Gastrointestinal disturbances were observed in 4 orally treated patients. A slight abnormal elevation of the glutamine-oxaloacetic transaminase level was also found in 1 patient orally treated. Since there were no statistically significant differences in efficacy rates between vaginal and oral CLDM treatments, we favor vaginal treatment of BV, based on less adverse effects.

Ovarian abscess caused by Peptostreptococcus magnus following transvaginal ultrasound-guided aspiration of ovarian endometrioma and fixation with pure ethanol.

BACKGROUND: Transvaginal ultrasound-guided aspiration of ovarian endometrioma has been applied and emphasized as a safe and simple procedure. CASE: Two 27-year-old infertile women, both gravida 0, para 0, underwent medical follow-up examinations for cases of ovarian endometrioma. Both had undergone transvaginal ultrasound-guided aspiration of ovarian endometrioma. Because both were continuously febrile and had abdominal pain and cysts with tenderness in spite of antibiotic therapies, both underwent laparotomies for treatment. In both cases, enucleation of the ovarian abscess revealed purulent and malodorous fluid that demonstrated Peptostreptococcus magnus in culture. CONCLUSION: We theorize that following transvaginal ultrasound-guided aspiration of ovarian endometrioma and fixation with pure ethanol, anaerobic infection by P. magnus occurred, and a cyst formed in the abscess.

Bacteriological epidemiology and treatment of bacterial vaginosis.

One hundred symptomatic women with clinical bacterial vaginosis (BV) were enrolled in this study. The bacteriological epidemiology of BV and efficacy of oral or vaginal administration of antimicrobial agents for treatment of BV were investigated. The epidemiology of BV was investigated before antimicrobial therapy. Fifty patients were treated with oral administration of metronidazole (MTN), 500 mg twice a day, and 50 patients with 2% clindamycin (CLDM) phosphate in a vaginal cream, 5 g once a day for 7 days. The major organisms occurring were Streptococcus agalactiae, Escherichia coli, Gardnerella vaginalis, Peptostreptococcus species, Bacteroides species, Prevotella species, and Mobiluncus species and opportunistic pathogens such as Staphylococcus epidermidis and Enterococcus faecalis. The therapeutic efficacy of CLDM cream on BV appeared to be preferable to oral MTN from clinical and bacteriological aspects. The clinical and bacteriological effectiveness of MTN and CLDM in the treatment of BV suggests that anaerobes might play a major role in causing the clinical symptoms in patients with BV.

Effects of crude herbal ingredients on intrauterine infection in a rat model

Abstract We investigated the effects of crude ingredients that compose Juzen-taiho-to (Shi-quan-da-bu-tang) and Ryutan-shakan-to (Long-dan-xie-gan-tang) using in vitro and in vivo experimental models. Juzen-taiho-to consists of 10 crude ingredients extracted from herbs: Astragali Radix, Glycyrrhizae Radix, Cinnamomi Cortex, Rehmanniae Radix, Paeoniae Radix, Cnidii Rhizoma, Atractylodis Lanceae Rhizoma, Angelicae Radix, Ginseng Radix, and Hoelen. Ryutan-shakan-to consists of 9 crude ingredients extracted from herbs: Scutellariae Radix, Glycyrrhizae Radix, Gardeniae Fructus, Rehmanniae Radix, Plantaginis Semen, Alismatis Rhizoma, Angelicae Radix, Akebiae Caulis, and Gentianae Scabrae Radix. For the in vitro experiment, the minimum inhibitory concentration of each extract of crude ingredient against Escherichia coli (GOG 0020) was > 100 μg/mL. For the in vivo experiment, 16 hours after inoculating rats with 8.1 × 10 6 colony-forming units of E coli , each extract of crude ingredient (125 mg/kg in 1 mL physiologic saline) orally three times a day or 1 mL physiologic saline alone (control) was administered to rats by gavage orally three times a day for 7 days. The extracts showed no in vitro antimicrobial activity, but extracts of Hoelen, Rehmanniae Radix, Glycyrrhizae Radix, Cinnamomi Cortex, Cnidii Rhizoma, Angelicae Radix, and Alismatis Rhizoma in both regimens significantly decreased the viable E coli counts in rat uterine infections.

In vitro Bactericidal Activities of Antimicrobial Agents and Morphologic Changes on Prevotella bivia

Prevotella bivia is common in pelvic inflammatory diseases. Parenteral antimicrobial agents have been widely used against those infections. We investigated the bactericidal activities of three cephalosporins, i.e. cefluprenam (CFLP), ceftazidime (CAZ) and cefotaxime (CTX) and of two other antimicrobial agents, i.e. clindamycin (CLDM) and imipenem (IPM) against P. bivia. We also investigated the in vitro morphological changes induced by these agents in P. bivia. Cephalosporins exhibited bactericidal activities against P. bivia and induced time- and concentration-dependent morphological changes in P. bivia (filamentation). CLDM and IPM also had bactericidal activities, but induced different morphologic alterations: formation of spheroblasts and lysis. These results confirm the fact that each antimicrobial agent has characteristic aspects.