Susceptibility to Fluconazole and Ketoconazole of Candida spp. Isolated from Primary and Episodic Vulvovaginites by E-Test (São Paulo, SP, Brazil)

Purpose: To evaluate the profile of in vitro susceptibility of yeasts isolated from cases of primary and espisodic vulvovaginitis to two antifungal agents. Methods: 40 Candida isolates from episodes of vulvovaginal candidiasis were identified by classic methodologies. The susceptibility testing of the in vitro fluconazole and ketoconazole activity against the isolates was accessed by E-test. Results: C. albicans was the most common species identified in 70% of the occurrences followed by C. glabrata (20%), C. tropicalis (7.5%), and C. guilliermondii (2.5%). In the susceptibility profile to antifungal agents, 12.5% and 16.7% of the isolates obtained from primary and episodic vulvovaginal candidiasis were resistant to fluconazole, respectively. To ketoconazole, we found that 6.25% and 12.5% of the isolates respectively from primary vulvovaginal candidiasis (PVVC) and episodic vulvovaginal candidiasis (EVVC) had high MIC values. Conclusions: E-test is a reliable method for the susceptibility testing of Candida spp. due to its simplicity, reproducibility, and lack of specialized equipment. Resistant strains and non-albicans species were verified more in cases of EVVC than in PVVC. Clinical and mycological cure of patients with episodic vulvovaginal candidiasis or complicated cases occurred after prolonged treatment and sometimes with multiple antifungals use.

<i>In Vitro</i>Effects of Photodynamic Therapy with Three Photosensitizers on <i>Candida</i>spp from Complicated Vulvovaginitis and Asymptomatic Women

Aim: To evaluate photodynamic therapy (PDT) in vitro to reduce the growth of Candida spp, and its synergy with the antifungals fluconazole and ketoconazole for inhibition of resistant, DDS and susceptible isolates from asymptomatic carriers and with complicated vulvovaginitis. Methods: Between 2017 and 2020, we evaluated 230 women with vulvovaginal candidiasis and 400 asymptomatic. We obtained 130 positive cultures for Candida spp from vulvovaginitis and 94 asymptomatic. Yeasts were characterized by classical and molecular tests. Sensitivity to fluconazole and ketoconazole was evaluated by E-test. We used photodynamic light through blue LED, wavelengths between 450 to 470 nm, power of 260 mW, energy fluence of 270 J/cm2, for 15 minutes over all colonies of Candida spp. Methylene blue (MB) at 450 mg/mL, 2% gentian violet (VG) and 50 μM curcumin (CR) were used in association or not with LED irradiation. Suspensions of Candida spp of 106 CFU/mL, subjected to the different assays, were introduced in 96-well microplates, incubated for 48 hours at 35˚C and the readings at 530 nm. The samples were finally cultivated in Petri plates containing Sabouraud dextrose agar to assess the growth inhibition. All procedures were in triplicate. Results: C. albicans was prevalent in vulvovaginal candidiasis, however, we also isolated non-albicans species such as C. glabrata, C. tropicalis and C. parapsilosis. There was a substantial reduction (66.6% to 83.8%) of the CFU/mL of the isolates treated with FDT. Gentian violet at 2% alone reduced the growth of CFU/ml of Candida spp from 69% to 75%. Among isolates of vaginitis and asymptomatic carriers, after using FDT, we found a reduction in resistant phenotypes and DDS for fluconazole in percentages from 20% to 100% for C. albicans, from 50% to 100% C. glabrata, 33.3% to 100% C. parapsilosis and 100% C. tropicalis. For ketoconazole in the same isolates, there was a reduction in phenotypes with MIC > 16 μg/mL of up to 50% in C. albicans, 50% to 100% C. glabrata, 50% to 100% C. tropicalis. Conclusions: PDT with MB, GV and CR revealed efficacy in vitro in reducing the growth of C. albicans and non-albicans, especially due to chronic recurrent vulvovaginitis.