Adhesion and biofilm formation of Candida parapsilosis isolated from vaginal secretions to copper intrauterine devices.

INTRODUCTION: Candida parapsilosis is one of the main species that is able to adhere to forming biofilms on inert materials. Adhesion is the first step towards the colonization and invasion of host cells during the infectious process. Among the infections, vulvovaginal candidiasis is increasingly common. The objective was to evaluate the profile of adherence and biofilm formation of eight isolates of C. parapsilosis on the metal used in intrauterine devices (IUDs). METHODS: Eight strains of C. parapsilosis presenting strong adhesion and biofilm formation properties were isolated from vaginal secretions in a previous study. To assay the adhesion and biofilm formation, copper fragments were made and cultivated in tubes containing 3 mL of phosphate-buffered saline and incubated for 6 and 24 h at 37 °C to evaluate biofilm formation. After incubation, the intensity of adherence and of biofilm formation on copper fragments were determined by performing a colony count. RESULTS: All isolates were able to form biofilms and the isolate Cp62 showed many cells joined in a planktonic mode forming biofilms. The use of an IUD is one of the main factors that favors vulvovaginal candidiasis, and the presence of copper in this device increases the chance of recurrent vulvovaginal candidiasis (CVVR) due to the ease with which species of the genus Candida can adhere to inert surfaces. CONCLUSION: This research showed that the clinical isolates studied adhered to IUD copper fragments and formed biofilms, further increasing their virulence.

Effects of intrauterine contraception on the vaginal microbiota.

OBJECTIVES: There have been conflicting reports of altered vaginal microbiota and infection susceptibility associated with contraception use. The objectives of this study were to determine if intrauterine contraception altered the vaginal microbiota and to compare the effects of a copper intrauterine device (Cu-IUD) and a levonorgestrel intrauterine system (LNG-IUS) on the vaginal microbiota. STUDY DESIGN: DNA was isolated from the vaginal swab samples of 76 women using Cu-IUD (n=36) or LNG-IUS (n=40) collected prior to insertion of intrauterine contraception (baseline) and at 6 months. A third swab from approximately 12 months following insertion was available for 69 (Cu-IUD, n=33; LNG-IUS, n=36) of these women. The V4 region of the bacterial 16S rRNA-encoding gene was amplified from the vaginal swab DNA and sequenced. The 16S rRNA gene sequences were processed and analyzed using the software package mothur to compare the structure and dynamics of the vaginal bacterial communities. RESULTS: The vaginal microbiota from individuals in this study clustered into 3 major vaginal bacterial community types: one dominated by Lactobacillus iners, one dominated by Lactobacillus crispatus and one community type that was not dominated by a single Lactobacillus species. Changes in the vaginal bacterial community composition were not associated with the use of Cu-IUD or LNG-IUS. Additionally, we did not observe a clear difference in vaginal microbiota stability with Cu-IUD versus LNG-IUS use. CONCLUSIONS: Although the vaginal microbiota can be highly dynamic, alterations in the community associated with the use of intrauterine contraception (Cu-IUD or LNG-IUS) were not detected over 12 months. IMPLICATIONS: We found no evidence that intrauterine contraception (Cu-IUD or LNG-IUS) altered the vaginal microbiota composition. Therefore, the use of intrauterine contraception is unlikely to shift the composition of the vaginal microbiota such that infection susceptibility is altered.

Does removal of CU-IUD in patients with biofilm forming candida really maintain regression of clinical symptoms?

OBJECTIVE: To evaluate whether symptoms and recurrence would differ with and without Cu-IUD removal in patients with concomitant biofilm forming Candida spp. METHODS: The data of 270 consecutive patients wearing TCu380A Cu-IUD were evaluated. Among these patients, 100/270 were found to have Candida spp. isolated from the tail of Cu-IUD or vaginal samples. These patients were investigated in four groups: Group 1 (n = 24; Biofilm (+), Cu-IUD removed), Group 2 (n = 14; Biofilm (+), Cu-IUD not removed), Group 3 (n = 29; Biofilm (-), Cu-IUD removed), Group 4 (n = 33; Biofilm (-), Cu-IUD not removed). Patients in each group were followed for clinical signs and symptoms for 8-16 months and compared to each other. RESULTS: Symptoms, physical findings and candida positivity have decreased statistically significantly in Group 1 one year after removal of Cu-IUD (95.8% vs. 4.2%, p < 0.01; 95.8% vs. 4.2%, p < 0.01; 100% vs. 8.3%, p < 0.01 respectively). In Group 2, symptoms, physical findings and candida positivity have decreased after follow-up, but without a statistical significance. In Group 3, all the parameters have decreased, but only decrease in candida positivity has reached statistical significance (100% vs. 48.3%, p < 0.01). In Group 4 - as in Group 1- symptoms, physical findings and candida positivity have decreased statistically significantly (48.5% vs. 18.2%, p = 0.01; 72.7% vs. 48.5%, p = 0.05; 100% vs. 51.5%, p < 0.01 respectively). CONCLUSION: Biofilm forming microorganisms should be considered in the management of vaginal infections or symptoms for safer use of intrauterine devices.

In vitro actinomycete biofilm development and inhibition by the polyene antibiotic, nystatin, on IUD copper surfaces.

The presence of intrauterine contraceptive devices (IUDs) gives a solid surface for attachment and an ideal niche for biofilm to form and flourish. Pelvic actinomycosis is often associated with the use of IUDs. Treatment of IUD-associated pelvic actinomycosis requires the immediate removal of the IUD. Therefore, this article presents in vitro evidence to support the use of novel antibiotics in the treatment of actinomycete biofilms. Twenty one clinical actinomycetes isolates from endocervical swabs of IUD wearers were assessed for their biofilm forming ability. An in vitro biofilm model with three isolates, Streptomyces strain A4, Nocardia strain C15 and Nocardia strain C17 was subjected to treatment with nystatin. Inhibition of biofilm formation by nystatin was found to be concentration dependent, with MBIC50 values in the range 0.08-0.16 mg ml(-1). Furthermore, at a concentration of 0.16 mg ml(-1), nystatin inhibited the twitching motility of the isolates, providing evidence for a possible mechanism of biofilm inhibition.

[In vitro biofilm formation and relationship with antifungal resistance of Candida spp. isolated from vaginal and intrauterine device string samples of women with vaginal complaints]. / Vajinal Yakinmasi Olan Kadinlarin Vajen ve Rahim Içi Araç Ipi Örneklerinden Izole Edilen Candida Türlerinin In Vitro Biyofilm Olusturma Özellikleri ve Antifungal Direnç ile Iliskisi.

Intrauterin device (IUD) application is a widely used effective, safe and economic method for family planning. However IUD use may cause certain changes in vaginal ecosystem and may disturb microflora leading to increased colonization of various opportunistic pathogen microorganisms. The aims of this study were (i) to detect the biofilm production characteristics of Candida spp. isolated from vaginal and IUD string samples of women with IUDs, and (ii) to investigate the relationship between biofilm production and antifungal resistance. A total of 250 women (mean age: 34.4 ± 7.6 years) admitted to gynecology outpatient clinics with vaginal symptoms (discharge and itching) were included in the study. The patients have been implanted CuT380a type IUDs for a mean duration of 59.8 ± 42.4 months. Without removing IUD, string samples were obtained by cutting and simultaneous vaginal swab samples were also collected. Isolated Candida spp. were identified by conventional methods and API 20C AUX (BioMerieux, Fransa) system. Minimal inhibitory concentrations (MIC) of fluconazole, itraconazole and amphotericin B were determined by broth microdilution method according to the CLSI guidelines. Biofilm formation was evaluated by crystal violet staining and XTT-reduction assays, and the isolates which yielded positive results in both of the methods were accepted as biofilm-producers. In the study, Candida spp. were isolated from 33.2% (83/250) of the vaginal and 34% (85/250) of the IUD string samples, C.albicans being the most frequently detected species (54 and 66 strains for the samples, respectively). The total in vitro biofilm formation rate was 25% (21/83) for vaginal isolates and 44.7% (38/85) for IUD string isolates. Biofilm formation rate of vaginal C.albicans isolates was significantly lower than vaginal non-albicans Candida spp. (14.8% and 44.8%, respectively; p= 0.003). Biofilm formation rate of C.albicans strains isolated from vaginal and IUD string samples were found as 14.8% (8/54) and 45.5% (30/66), with a statistically significant importance (p< 0.001). However, no statistically significant difference was detected for biofilm formation rates of non-albicans Candida spp. when sample types were considered [44.8% (13/29) and 42.1% (8/19), respectively; p> 0.05]. Fluconazole resistance was significantly higher in biofilm-producing vaginal Candida spp. than those of nonproducers (52.4% vs. 16.1%; p= 0.001), however, itraconazole resistance was found similar in biofilmproducer and non-producer isolates (47.6% vs. 32.3%; p> 0.05). Resistance rates for both fluconazole and itraconazole were higher in biofilm-producers (39.5% and 52.6%, respectively), than those of non-producers (10.6% and 29.8%, respectively), representing a statistical significance (p= 0.002 and p= 0.03, respectively) for Candida spp. strains isolated from IUD string samples. The overall resistance rates of C.albicans and non-albicans Candida spp. against fluconazole, were determined as 15% and 54.2%, respectively, while those rates were 24.2% and 68.7%, respectively, against itraconazole. MIC value of amphotericin B for all of the Candida spp. isolates was ≤ 1.5 µg/ml. In conclusion, the data obtained from this study revealed that Candida spp. May lead to vaginal infections by inducing biofilm formation in IUD strings and these biofilms may be related to resistance to antifungal agents. Thus, women using IUDs should be followed-up periodically for the development of biofilms in their IUD strings.

In vitro Actinomyces israelii biofilm development on IUD copper surfaces.

BACKGROUND: Female pelvic actinomycosis may involve fallopian tubes, ovaries, uterus and bladder. This condition is often associated with the use of intrauterine contraceptive devices (IUDs), vaginal pessaries and/or tampons. The predominant causative agent of human actinomycosis is Actinomyces israelii, which has been found on copper IUDs retrieved from patients. STUDY DESIGN: In this work, a biofilm of A. israelii was developed in vitro on copper surfaces immersed in a simulated uterine fluid under anaerobic conditions. The biofilm was characterized using scanning electron microscopy (SEM), energy dispersive X-ray and atomic force microscopy. RESULTS: The capacity of A. israelii to develop a biofilm over copper surfaces in synthetic media was demonstrated. SEM micrographies illustrate the exopolysaccharides production and bacterial distribution. CONCLUSION: A. israelii was able to attach and grow in synthetic intrauterine media and to present on the copper surface is likely due to the production of biofilm.

Can intrauterine contraceptive devices be a Candida albicans reservoir?

BACKGROUND: The in vitro adherence of Candida albicans isolated from vaginal exudates of patients with vulvovaginal candidiasis (VVC) to intrauterine contraceptive devices (IUDs) and biofilm formation capacity were evaluated. STUDY DESIGN: This research was conducted with two vaginal C. albicans isolates. The adherence on IUD by both radiomarked adhesion assay and scanning electron microscopy, and the biofilm production capacity by spectrophotometric method were determined. RESULTS: The yeasts adhered strongly to different parts of the IUD (covered with copper wire, without copper wire and tail), and there was no significant difference in the rates of adhesion to the different parts (p=.7771). The vaginal yeasts showed a high capacity to produce biofilm. CONCLUSIONS: Two vaginal yeasts evaluated showed a high capacity to produce biofilm on IUD. It was confirmed that all parts of the IUD allow the adherence of yeasts. The adherence of C. albicans to different parts of the IUD and its formation of biofilm seems to be important attributes influencing the occurrence of VVC and recurrent VVC.

Tubal pathology: the role of hormonal contraception, intrauterine device use and Chlamydia trachomatis infection.

BACKGROUND: The aim of the present study was to identify a possible association between tubal pathology and the contraceptive methods previously used, especially third generation copper intrauterine devices (IUDs). METHOD: In this retrospective unmatched case-control study, use of contraceptive methods and Chlamydia trachomatis antibody titres were compared in women with tubal occlusion (n = 84) and infertile women with tubal patency (n = 253) demonstrated by hysterosalpingography. Contraceptive methods were also compared to primiparous women with natural pregnancy. RESULTS: A significantly higher percentage of women with tubal occlusion had previously used an IUD. Positive C. trachomatis antibody testing was found significantly more often in women with tubal occlusion. Neither hormonal contraceptives nor condoms were associated with an increased risk for uni- or bilateral tubal occlusion. CONCLUSION: In the present study, we observed an increased rate of tubal occlusions in previous IUD users. This should be considered in the contraceptive counseling of young and nulliparous women.

Reactive oxygen intermediates and reactive nitrogen intermediates in copper intrauterine device users.

The mechanism of action of copper in copper intrauterine devices (Cu IUD) as an antimicrobial agent is not well understood. Copper and iron are supposed to be responsible for release of reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI), which are very active in the presence of infection. The copper in a copper IUD could be responsible for limiting pelvic inflammatory disease. The present study was composed of 20 IUD seekers in whom ROI and RNI were studied before insertion of Cu IUD and then at 1, 4, and 12 weeks afterward. ROI showed a rise after insertion, whereas RNI showed a steady decline. Hence, it is presumed that the rise in ROI could be responsible for both the bactericidal effect of Cu IUD and also for the fall in RNI.

PIP: There is evidence of an increase in the number of polymorphonuclear leukocytes in the endometrium after insertion of a copper IUD. When activated, these macrophages secrete two groups of toxic metabolites that interact with each other: reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI). The mechanism of action of the copper in copper IUDs as an antimicrobial agent is not well understood. The present study measured ROI and RNI levels in 20 copper IUD users from Chandigarh, India, before and 1, 4, and 12 weeks after insertion. A statistically significant decrease in RNI levels was observed at each postinsertion time. In contrast, ROI levels showed an initial fall at 1 week postinsertion followed by a rise at 4 and 12 weeks. Control of infection has been found to be associated with this pattern of a rise in ROI and fall in RNI. (The initial fall in ROI at 1 week observed in the present study is assumed to be an anomaly related to the introduction of microbials during IUD insertion.) The copper in a copper IUD could be responsible for limiting pelvic inflammatory infection.