Vulvovaginal Candidiasis Patients Research Articles

Genetic diversity and molecular epidemiology of Candida albicans from vulvovaginal candidiasis patients.

Candida albicans (C. albicans) is a common cause of vulvovaginal candidiasis (VVC). In this paper, the genetic diversity and molecular epidemiology of 173C. albicans strains were investigated by multilocus sequence typing (MLST). A total of 52 diploid sequence types (DSTs) were recognized, and 27 (51.9%) of which have not been reported in the MLST database. Genotyping was performed on the multiple isolates collected from patients with recurrent VVC (RVVC, referring to VVC which attacks more than 4 times in one year) in different acute infectious phases. The results showed that 59.1% (26/44) of the patients suffered a relapse, with DST 79 (65.4%) as the dominant genotype. The etiology of the remaining 40.9% (18/44) of patients was reinfection, and the main genotypes included DST 79 (33.3%), DST 124 (8.6%) and DST 1895 (8.6%). DST 79 (45%) and DST 1395 (7.5%) were the main isolates of VVC patients, while DST 79 (24.1%), DST 727 (6.9%), DST 732 (6.9%) and DST 1867 (6.9%) were the main types of healthy volunteers. The results of the genotypes between RVVC patients and other groups were statistically different. Furthermore, cluster analysis was carried out on 1468 isolates, among which 1337 were downloaded from the MLST database, 130 were divided into 8 Clades in the present study and the remaining one was taken as a singleton. 92.3% isolates from relapse patients, 58.3% isolates from re-infected patients, 77.5% isolates from VVC patients and 51.7% isolates from volunteers were distributed in Clade 1. The analysis of the genotypes of multiple isolates from RVVC patients further demonstrated that point mutation and loss of heterozygosity contributed to the microevolution of C. albicans.

Molecular Investigation of Etiologic Agents Causing Vulvovaginal Candidiasis

Background: Vulvovaginal candidiasis (VVC) is an ordinary infection caused by Candida species. Meanwhile, a shift towards non-albicans Candida (NAC) species has been detected in VVC patients. Objectives: This study aimed at molecular identification of Candida isolates, causing VVC. Methods: Vaginal secretion samples of 320 non-pregnant vaginitis patients at Shahid Akbar-Abadi Obstetrics and Gynecology Hospital in Tehran (Iran) were collected. Samples were evaluated using mycological and molecular approaches. Vaginitis isolates were analyzed with the PCR using NL1 and NL4 primers, and the D1/D2 region of the large-subunit rRNA gene was amplified and sequenced. Results: In total, 100 Candida isolates were identified from VVC and recurrent vulvovaginal candidiasis (RVVC). Candida albicans was the most frequent (51%), followed by C. glabrata (36%), C. krusei (Pichia kudriavzevii) (8%), and C. kefyr (Kluyveromyces marxianus) (5%). 51 and 49% of isolates had C. albicans and NAC, respectively. Conclusions: Candida albicans and C. glabrata were the most common agents of vulvovaginal candidiasis. NAC spp. (49%) was found as an important agent associated with VVC.

Antifungal Activity and Mode of Action of Miltefosine Against Clinical Isolates of Candida krusei.

Candida krusei attracts attention from medical professionals mainly for its intrinsic resistance to fluconazole and the limited number of drugs available to treat C. krusei vulvovaginal candidiasis. Miltefosine was demonstrated to have good antifungal activity both in vitro and in vivo. Here, we determined the susceptibility profiles of 57 clinical C. krusei isolates from vulvovaginal candidiasis patients and assessed the antifungal activity of miltefosine against C. krusei. All isolates were susceptible to voriconazole and itraconazole, whereas 1.8% of the isolates were of non-wild-type phenotype to amphotericin B. In contrast, miltefosine showed low MICs against all C. krusei isolates with fungicidal activity. The checkerboard assay showed that the synergistic effect of miltefosine in combination with amphotericin B was observed in 25% of the tested planktonic C. krusei isolates and 18.8% of the tested preformed biofilms, whereas miltefosine in combination with fluconazole showed indifferent interaction for all tested planktonic isolates. The presence of sorbitol in the broth microdilution assay did not influence the MIC values of miltefosine against C. krusei, but the presence of ergosterol increased the MIC values. Visible changes in cell content in cells treated with miltefosine were observed. We found that cells treated with miltefosine showed decreased cell viability and chromatin condensation under PI staining, which indicates that miltefosine may induce apoptosis-like cell death in C. krusei. In conclusion, we found miltefosine has a good activity against C. krusei isolates and exerts its fungicidal effect by binding to ergosterol in the cell membrane and inducing apoptosis.

Study on Vulvovaginal Candidiasis: Clinical Epidemiology and <i>in vitro</i> Susceptibility of Pathogenic Yeasts in China

Background: Vulvovaginal candidiasis (VVC) is a common vaginal infection. We aim to investigate the clinical epidemiology of VVC and the in vitro antifungal susceptibility of Candida spp. collected from different regions of China, so as to provide a guideline for the appropriate use of antifungals in clinical settings. Methods: Candida isolates from VVC patients and questionnaires were collected from 12 hospitals across the China from September 2017 to March 2018. Species were identified by germ tube test, clamydospore formation, CHROMagar medium and DNA sequencing. Susceptibility to fluconazole (FLC), itraconazole (ITC), voriconazole (VOC), posaconazole (POS), miconazole (MIC), econazole (ECO), clotrimazole (CLZ), nystatin (NYS), amphotericin B (AMB), caspofungin (CAS), and terbinafine (TBF) was determined using the CLSI broth microdilution method (M27-E4). The in vitro data were also evaluated with updated clinical breakpoints and epidemiological cut-off values. Findings: A total of 602 cases were enrolled in this study, of which 253 (42%) were uncomplicated VVC and 349 (58%) were complicated. Among complicated VVC, 26% were RVVC. The subjects were between 16 and 64 years of age, of which 60% were between 25 and 35 years of age. A total of 543 strains were isolated from these VVC patients. The most commonly identified species was C.albicans (84.71%), followed by C.glabrata (8.66%), Meyerozyma caribbica (1.29%), Pichia guilliermondii (1.29%), C.tropicalis (0.92%), Pichia kudriavzevii (0.92%), C.parapsilosis (0.55%), Candida nivariensis (0.55%), Issatchenkia orientailis (0.55%) and less common yeasts(0.55%). The overall susceptibility rates of C.albicans to FLC, VOC and CAS were all greater than 80%, and the resistance rates were less than 10%. Most of C.albicans isolates were WT to ITC (86%), POS (95%) and AMB (98%). Most of C.glabrata (98%) were dose-dependently susceptible to FLC. All of C.glabrata isolates were WT to both ITC and AMB, while most were WT to both VOC (85%) and POS (91.5%).The proportion of NWT strains to FLC in RVVC was much higher than that in other types of VVC(33% vs . 23%, P <0.05). The proportions of WT strains to ITC in complicated VVC and RVVC were all over 80%, which were much higher than that to FLC( P <0.01). CLZ showed the lowest MIC 90 (0.06ug/ml)against C.albicans, and the MICs of CLZ among non-albicans Candida (NAC) were all <2ug/ml. Interpretation: VVC was more frequent in women of childbearing age. C.albicans (84.71%) was still the predominant species of VVC in China, while C.glabrata (56.6%) was the main species in NAC. Most C.albicans strains isolated from uncomplicated VVC in different regions of China were still susceptible to the commonly used azoles, including FLC and ITC, which could be used as empirical treatment. However, FLC may not be the first choice for the treatment of complicated VVC, especially RVVC. In both cases, ITC was more appropriate treatment. CLZ could be recommended as a good choice of topical drugs for VVC. As for new drugs, POS is promising for the refractory cases. Funding Statement: This study was supported by National Science and Technology Major Project (2018ZX10734404), the CAMS Initiative for Innovative Medicine (2016-I2M-3-021), National Natural Science Foundation of China (Grant No. 81573059), the Basical Scientific Research Fund Projects of Chinese Academy of Medical Sciences (No. 2018PT31013) and Nanjing Clinical Medicine Research Center Project (No.2019060001). Declaration of Interests: All authors declare that they have no conflicts of interest. Ethics Approval Statement: The study has been approved by the ethics committee of the hospital, and all patients have signed informed consent (version number and date:1.0, 2017.02.16).

A Role for Yeast/Pseudohyphal Cells of Candida albicans in the Correlated Expression of NLRP3 Inflammasome Inducers in Women With Acute Vulvovaginal Candidiasis.

In acute vulvovaginal candidiasis (VVC), the fungus Candida albicans activates inflammasome receptors of vaginal epithelial cells through the production of virulence and immuno-inflammatory factors. Here, we show that in VVC patients, genes encoding some of the above factors (SAP2, SAP5, SAP6, ECE1, and HWP1) are expressed in a correlated fashion. Cytological observations pointed out that pseudohyphal filaments with yeast cells are dominant at the acidic vaginal pH, and this is coupled with co-expression, at roughly similar level, of SAP2, a typical yeast and ECE1, a typical hyphae-associated genes. In contrast, vigorous hyphal growth dominated at the neutral vaginal pH of mice experimentally infected with C. albicans isolates from VVC subjects, and this is coupled with a high ratio of ECE1 to SAP2 expression. We suggest that the pseudohyphal rather than true hyphal cells of C. albicans play a critical role in VVC, possibly through the activity of multiple inflammasome inducers.

Regulatory Role of ERG3 and Efg1 in Azoles-Resistant Strains of Candida albicans Isolated from Patients Diagnosed with Vulvovaginal Candidiasis.

Vulvovaginal candidiasis (VVC), caused by Candida albicans, affects women's health and life. We aimed to explore the correlation between ERG3 as well as Efg1 mutation/overexpression and azoles-resistance, and the correlation between ERG3 and Efg1 mRNA expression in C. albicans. First, C. albicans was isolated from clinical VVC patients. ERG3 and Efg1 mutations were detected by polymerase chain reaction (PCR) and sequencing, and the expression levels of these two genes were also identified by qRT-PCR. Correlations between mutation/overexpression of ERG3/Efg1 and azoles-resistance as well as ERG3 and Efg1 mRNA expression were analyzed. Based on the ERG3 sequencing, the results showed that there were 2 missense mutation sites, 1 nonsense mutation site, and 4 silent mutation sites, while 1 missense mutation sites, 1 nonsense mutation site, and 12 silent mutation sites were found in Efg1. Furthermore, the mRNA levels of ERG3 gene in the strains sensitive to FCA, ITR or VRC were higher than those in the strains resistant to FCA, ITR, VRC (P < 0.05). While for the mRNA levels of Efg1, susceptible strains were lower than resistant strains. Besides, there was a significant linear negative correlation between ERG3 and Efg1 mRNA expression (r = - 0.614, P < 0.001).

Azole Resistance and Detection of the ERG11 Gene in Clinical Candida albicans Isolated from Pregnant Women with Vulvovaginitis Attending Federal Medical Centre, Yenagoa, Nigeria

Introduction: Candida albicans is one of the most important aetiological agents causing vaginal candidiasis in pregnant women. Most women will experience at least one episode during their reproductive years. Antifungal resistance is a particular problem with Candida infections. Some types of Candida are increasingly resistant to the first-line and second-line antifungal medications.&#x0D; Objective: To investigate the azole susceptibility of Candida albicans (C. albicans) from pregnant vulvovaginal candidiasis patients and to detect ERG11 gene in these azole resistance isolates.&#x0D; Methods: Forty-one clinical isolates of C. albicans were collected. Azole susceptibility was tested in vitro using microdilution techniques. The ERG11 genes of 27 isolates of C. albicans (All resistant to azoles) were amplified using PCR method.&#x0D; Results: Of the 67 isolates recovered, 41(61.19%) were C. albicans, of which 27 (65.85%) each, and 25(60.98%) were resistant to Fluconazole, Voriconazole, and Nystatin respectively. In total, ERG11 genes were detected among 24(88.89%) of 27 C. albicans azole resistant isolates.&#x0D; Conclusions: Twenty four ERG11 genes were detected among 27 azole resistant C. albicans isolates, which indicates a possible relation with the increase in resistance to azole drugs and the recurrence of vulvovaginal candidiasis.

Differential expression levels of agglutinin-like sequence, lipase, and secreted aspartyl protease genes in Candida tropicalis treated with fluconazole alone and in combination with clotrimazole

Background: The frequency of opportunistic fungal infections in immunocompromised patients, especially by Candida species, has sharply increased in the last few decades. As the number of antifungal drugs available for the treatment of candidiasis is limited, combination therapy has been employed as one of the most commonly used techniques to alleviate this problem. Aims: The main aim of this study was to explore the antifungal activity of fluconazole in combination with clotrimazole on expression levels of virulence genes, agglutinin-like sequences (ALS1 and ALS2), lipases (LIP1 and LIP4) and secreted aspartyl proteases (SAP2 and SAP4) in Candida tropicalis. Methods: Ten infected clinical isolates obtained from recurrent vulvovaginal candidiasis patients were used in this study. The broth microdilution assay was utilized to investigate antifungal susceptibilities to fluconazole alone and in combination with clotrimazole and the synergistic effects were interpreted with reference to the fractional inhibitory concentration (FIC) index model. The expression levels of ALS1, ALS2, LIP1, LIP4, SAP2 and SAP4 genes were quantified by real-time RT–PCR. Results: Antifungal susceptibility results showed that isolates were resistant to at least one type of azole antifungals. The combination of fluconazole with clotrimazole revealed synergistic effects against C. tropicalis isolates with FIC90 index ranging from 0.011 to 0.43. The results indicated that combination of fluconazole with clotrimazole could cause a down-regulation of gene expression of ALS1, SAP2, LIP4, SAP4, LIP1 and ALS2 genes, respectively. Conclusions: Fluconazole in combination with clotrimazole may diminish the virulence properties of C. tropicalis.

Identification of Candida Species Isolated from Vulvovaginal Candidiasis Patients by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) in Yasuj Southwestern Iran

Background: Vulvovaginal candidiasis (VVC) is a frequent infection in females at the reproductive age. Furthermore, the most common causative agent is Candida albicans yet in the recent years, the incidence of non-albicans species has increased. Objectives: The main aim of this study was the isolation and identification of various Candida species from patients with vulvovaginal candidiasis by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in Yasuj, Iran. Methods: Three hundred and ten suspected women females vaginitis were sampled and examined. Genomic DNA was extracted from fresh colonies by phenol-chloroform glass bead methods. Polymerase chain reaction (PCR) amplification was performed based on the ribosomal DNA internal transcribed spacer (rDNA-ITS), and specific electrophoretic patterns of PCR products after digestion with MspI enzyme used for species identification. Results: The cultures were positive for 160 (51.6%) vaginal samples. Candida albicans (86.8%) was the most common species among the isolates, followed by C. glabrata (3.77%) and C. krusei (3%). Multispecies with two Candida were identified in nine patients. Conclusions: Vulvovaginal candidiasis is more prevalent among females in Yasuj and the predominant agent is C. albicans. In addition, correct identification of Candida species could play an important role in management and treatment of VVC.

Risk Factors of Vulvovaginal Candidiasis among Women of Reproductive Age in Xi'an: A Cross-Sectional Study.

Purpose. To explore risk factors of vulvovaginal candidiasis (VVC) among women of reproductive age in Xi'an district and then to offer reference for clinical prevention and treatment of VVC. Methods. Patients from the outpatient department of gynecology and obstetrics in the First Affiliated Hospital of Xi'an Jiaotong University from June 2016 to May 2017 were recruited strictly according to the inclusion and exclusion criteria. Participants diagnosed as simple VVC were assigned to the case group, while women who underwent routine gynecological examination and had normal vaginal microflora were assigned to the control group. Then we conducted a questionnaire survey of the two groups and used the logistic regression model to explore the related risk factors of VVC. Results. In the present study, ninety-seven cases were sample VVC patients and eighty-seven cases were healthy women. This cross-sectional study showed that occasionally or never drinking sweet drinks (odds ratio [OR] =0.161, 95% confidence interval [CI] =0.056-0.462, P=0.001), occasionally or never eating sweet foods (OR=0.158, 95%CI=0.054-0.460, P=0.001), and the use of condom (OR=0.265, 95%CI=0.243-0.526, P=0.001) were regarded as protective factors for VVC. In addition, sedentary life style (OR=7.876, 95%CI=1.818-34.109, P=0.006), frequently wearing tights (OR=6.613, 95%CI=1.369-27.751, P=0.018), frequent intravaginal douching (OR=3.493, 95%CI=1.379-8.847, P=0.008), having the first sexual encounter when under 20 years old (OR=2.364, 95%CI=1.181-7.758, P=0.006), the number of sexual partners being over two (OR=3.222, 95%CI=1.042-9.960, P=0.042), history of curettage (OR=3.471, 95%CI=1.317-9.148, P=0.012), history of vaginitis (OR=8.999, 95%CI=2.816-28.760, P<0.001), and not cleaning the vulva before or after sexual encounters (OR=13.684, 95%CI=2.843-65.874, P=0.001) were considered to be risk factors of VVC. Conclusion. In conclusion, risk factors of VVC are various, involving ages, hygienic habits, disease history, and other aspects.

RISK FACTORS OF VULVOVAGINAL CANDIDIASIS IN DERMATO-VENEREOLOGY OUTPATIENTS CLINIC OF SOETOMO GENERAL HOSPITAL, SURABAYA, INDONESIA.

Background:Vulvovaginal Candidiasis (VVC) is one-third of vaginitis case. About 75% of women will have at least one episode of VVC. Complication of VVC can be unfavorable to impact the patient’s quality of life. Knowing its risk factors can prevent someone suffering from pathological VVC and its sequelae. The purpose of this study was to identify risk factors of VVC.Materials and Methods:A descriptive retrospective study was conducted by total sampling to 213 medical records of VVC patients from 869 fluor albus patients in Sexually Transmitted Infection (STI) Division, Dermatology and Venereology Outpatient Clinic in Dr. Soetomo General Hospital, Surabaya, East-Java, Indonesia in 2011 to 2013.Results:In 2011, 69 cases (22.77%) from 303 fluor albus patients, in 2012 69 cases (22.69%) from 304 fluor albus patients, and in 2013 75 cases (28.63%) from 262 fluor albus patients. As many as 180 (84.50%) of the patients were sexually active age group, which was 15-44 years old. Mostly patients were married (69.48%). As many as 167 (78.40%) sexual partners were patient’s husband. About 95 (44.13%) patients presented with recurrent VVC. Risk factors are previous STIs 74 (34.74%), vaginal douching (27.70%), pre-martial sexual intercourse (5.63%), other STIs (3.29%), diabetes mellitus (2.34%), gestation (1.88%), other fluor albus infections (0.94%), pantyliner (0.47%), and STIs in sexual partner (0,47%).Conclusion:Avoiding and/or managing risk factors is important to prevent VVC and its complications.

Molecular Identification and Antifungal Susceptibility Pattern of Non-albicans Candida Species Isolated from Vulvovaginal Candidiasis

Background:Vulvovaginal candidiasis (VVC) is an important health problem caused by Candida spp. The aim of this study was molecular identification, phylogenetic analysis, and evaluation of antifungal susceptibility of non-albicans Candida isolates from VVC.Methods:Vaginal secretion samples were collected from 550 vaginitis patients at Sayyad Shirazi Medical and Educational Center of Gorgan (Golestan Province, Iran) from May to October 2015. Samples were analyzed using conventional mycological and molecular approaches. Clinical isolates were analyzed with specific PCR using CGL primers, and the internal transcribed spacer region and the D1-D2 domain of the large-subunit rRNA gene were amplified and sequenced. Susceptibility to amphotericin B, fluconazole, itraconazole, and clotrimazole was determined by the guidelines of the Clinical and Laboratory Standard Institute.Results:In total, 35 non-albicans Candida isolates were identified from VVC patients. The isolates included 27 strains of Candida glabrata (77.1%), 5 Candida krusei (Pichia kudriavzevii; 14.3%), 2 Candida kefyr (Kluyveromyces marxianus; 5.7%), and 1 Candida lusitaniae (Clavispora lusitaniae; 2.9%). The fungicides itraconazole and amphotericin B were effective against all species. One isolate of C. glabrata showed resistance to fluconazole and clotrimazole, and 26 isolates of C. glabrata indicated dose-dependent susceptibility to fluconazole. C. lusitaniae was susceptible in a dose-dependent manner to fluconazole and resistant to clotrimazole.Conclusions:Non-albicans Candida spp. are common agents of vulvovaginitis, and C. glabrata is the most common species in the tested patients.

Patient Preferences in the Treatment of Vaginal Candidiasis

Vulvovaginal candidiasis (VVC) is one of the most frequent problems facing women and their healthcare professionals (HCP). It is difficult for providers to understand the prevalence of VVC because effective over the counter treatments (OTC), are available. It is expected that there are a great many more episodes of VVC in our patient population, as the frequency of self-treatment, success and satisfaction for those that use OTC products prior to seeing a women’s health care provider is unknown. In this study; healthcare providers were given OTC miconazole/tioconazole units for free distribution to patients for whom they diagnosed VVC by exam in their offices. Surveys for both HCP and patients were also distributed to determine the initial or recurrent nature of their symptoms and their satisfaction with their treatments. 1265 OBGYNs and 1821 NP/CNM/PAs reported on over 19,000 patients receiving a single complete treatment. Among HCPs, treatment efficacy was the primary reason for recommending miconazole/tioconazole. However, rapid onset of symptom relief, safety vs. fluconazole, patient preference for the less messy ovule, avoidance with drugs metabolized by the liver, and resistance to, or failure on, fluconazole were additional reasons noted for topical preference. Overall, patient satisfaction (n = 317) was high at 95%, with 97% of patients reporting that they would use miconazole again. Importantly, 100% of first time sufferers (n = 95) would use miconazole again. The few patients dissatisfied with miconazole (2%), reported no symptom relief, burning after use, an “allergic reaction”, pain from the applicator or a messy discharge. Most patients using the external symptom relief products were satisfied with the external cream (93%) and wipes (96%). Almost all patients reported their yeast infection was cured (91%) and that they experienced quick symptom relief (94%), despite the prevalence of recurrent vs first time VVA sufferers that presented to offices in this trial. Most patients were cured and satisfied with their treatment, therefore supporting the use of topical miconazole/tioconazole as first line therapy for vulvovaginal candidiasis patients presenting to a healthcare office.

Antifungal susceptibility and test for cure of &lt;i&gt;candida species&lt;/i&gt; among vulvovaginal candidiasis patients in a secondary care hospital, Nigeria

Background: Antimicrobial resistance among Candida species is an intense public health concern. The aim of the study was to determine the antifungal susceptibility pattern and test for cure of Candida species among women of child bearing age who visited the General Hospital Onitsha, Nigeria with symptoms suggestive of Vulvovaginal Candidiasis (VVC). Materials and Methods: Eight hundred and seventy six female patients participated in the study of which high vaginal swabs were collected and evaluated mycological by standard microbiological methods: microscopic examination and culture using sabouraud dextrose agar (SDA). Susceptibility of isolates to 4 antifungal agents was tested using agar dilution method. Clinicomycological evaluation was also performed among the patients. Result : Higher minimum inhibitory concentration (MIC) to azole antifungals was observed predominantly among non-albicans Candida species increasingly involved in VVC. The rate of mycological resolution was higher than symptomatic relief at 2 weeks after treatment with antifungal drug. Conclusion : Efficacious treatment of VVC requires an adequate knowledge of the causative agents and more importantly the antimicrobial to which they exhibit high susceptibility. Keywords: Vulvovaginal Candidiasis, Clinico- mycology, Antimicrobial resistance, Candida species Susceptibilite antifonique et test pour la cure d'especes de candida entre les patients de candidases vulvovaginales dans un hopital de soins secondaires, Nigeria Contexte: La resistance aux antimicrobiens chez les especes de Candida est un probleme de sante publique intense. L'objectif de l'etude etait de determiner le schema de susceptibilite aux antifongiques et le test de guerison des especes de Candida parmi les femmes en âge de procreer qui ont visite l'hopital general de Onitsha, au Nigeria, avec des symptomes suggerant une candidose vulvovaginale (VVC). Materiaux et methodes: huit cent soixante-seize six patientes ont participe a l'etude des prelevements vaginaux eleves collectes et evalues par mycologie par methodes microbiologiques standard: examen microscopique et culture a l'aide de la gelose sabouraud dextrose (SDA). La susceptibilite des isolats a 4 agents antifongiques a ete testee en utilisant une methode de dilution en agar. Une evaluation clinico-mycologique a egalement ete realisee chez les patients. Resultat: une concentration minimale minimale d'inhibition (MIC) en anatoxines azoliques a ete observee principalement chez les especes non-albicans Candida de plus en plus impliquees dans VVC. Le taux de resolution mycologique etait plus eleve que le soulagement symptomatique a 2 semaines apres le traitement par un medicament antifongique. Conclusion : Un traitement efficace de la VVC necessite une connaissance adequate des agents causaux et, plus important encore, des antimicrobiens auxquels ils presentent une forte susceptibilite. Mots-cles : Candidiase Vulvovaginale, Clinico-mycologie, Resistance Antimicrobienne, Especes Candida

Clonal spread and azole-resistant mechanisms of non-susceptible Candida albicans isolates from vulvovaginal candidiasis patients in three Shanghai maternity hospitals.

In our multicenter study, 43 fluconazole non-susceptible and 45 fluconazole-susceptible isolates were collected from vulvovaginal candidiasis (VVC) patients from three Shanghai maternity hospitals to analyze their molecular epidemiological features and fluconazole resistant mechanisms. Cross-resistance to fluconazole, itraconazole and voriconazole was observed in 53.5% of the nonsusceptible isolates. Though we acquired 12 clonal complexes (CCs) of diploid sequence types (DSTs) in clinical isolates by a multilocus sequence typing method, fluconazole nonsusceptible isolates all belonged to CC69 with a predominant genotype of DST 79. Increased expressions of efflux pump genes (CDR1, CDR2, and MDR1) were observed only in minor fluconazole non-susceptible isolates by real-time quantitative polymerase chain reaction (PCR). However, ERG11 genes of fluconazole SDD and resistant isolates had significantly higher expression levels than fluconazole-susceptible isolates. Moreover, 13 distinct amino acid substitutions in Erg11p were found in clinical isolates. Three of the substitutions were novel amino acid substitutions (T123I, P98S, and Y286D), which were not in the susceptible isolates. Only two heterozygous amino acid substitutions (A18P/A and R365G/R) in Erg3p were found in two isolates with cross-resistance to fluconazole, itraconazole, and voriconazole. Taken together, we observed the clonal spread of CC69 in fluconazole non-susceptible isolates of Candida albicans from VVC patients with the dominant genotype DST79. ERG11 gene mutations and overexpression predominantly contributed to fluconazole resistance instead of the more common increased expressions of efflux pump genes (CDR1, CDR2, and MDR1).

Effect of chloroform extracts from Longdan Xiegan decoction in inhibiting hydrolytic enzyme activity of Candida albicans isolated from VVC patients

To investigate the inhibitory effect and mechanism of chloroform extracts from Longdan Xiegan decoction(CELX) against hydrolytic enzymes activity of Candida albicans isolated from vulvovaginal candidiasis(VVC) patients. Secreted aspartyl proteinase(Sap), phospholipase(PL) and lipase(Lip) positive strains were identified from 15 strains of C. albicans with milk culture medium, egg yolk culture medium and tween-80 medium, respectively. Then, the activities of Sap, PL, and Lip were detected in the above media. qRT-PCR was used to detect the changes in gene expressions of aspartic protease(SAP1-7,10), phospholipase B(PLB1-2) and lipase(LIP3-6). Secreted aspartyl proteinase and phospholipase of 15 VVC clinical strains were positive, and lipase of 11 strains were positive. Compared with the blank control group, the drug CELX-containing medium(milk medium, egg yolk culture medium, tween-80 medium) experiment showed that the sedimentation of colonies decreased gradually in each culture medium with the increase of CELX dose. When the concentration of CELX was 256 mg•L⁻¹, the colony almost disappeared, which indicated the enzyme activity was significantly weakened. The results of qRT-PCR showed that SAP1, SAP2, SAP3, SAP4, SAP7, SAP9 and SAP10 were down-regulated by 62%, 55%, 62%, 84%, 61%, 51%, 68%, respectively, except for SAP5 and SAP6; and PLB1, LIP3, LIP4, LIP6 were down-regulated by 67%, 51%, 54%, 55%, respectively. The findings suggested that CELX may inhibit the activities of Sap, PL, and Lip, which are important virulence factors of C. albicans.

Biofilms Formed by Isolates from Recurrent Vulvovaginal Candidiasis Patients Are Heterogeneous and Insensitive to Fluconazole.

Vulvovaginal candidiasis (VVC) is a global health problem affecting ∼75% of women at least once in their lifetime. Here we examined the epidemiology of VVC in a patient cohort to identify the causative organisms associated with VVC. Biofilm-forming capacity and antifungal sensitivity profiles were also assessed. We report a shifting prevalence of Candida species with heterogeneous biofilm-forming capacity, which is associated with altered antifungal drug sensitivity.

Efficacy of vitamin B complex as an adjuvant therapy for the treatment of complicated vulvovaginal candidiasis: An in vivo and in vitro study.

This study aimed to explore the efficacy of vitamin B complex as an adjuvant therapy for the treatment of complicated vulvovaginal candidiasis (VVC) in vitro and in vivo. One-hundred fifty-eight complicated VVC patients were randomly divided into group A (treated with suppository+oral antifungal agents), group B (treated with suppository+vaginal cream), and group C (treated with suppository+vaginal cream+oral vitamin B complex). A mouse model of VVC was established. Eighty VVC mice were randomly divided into 4 groups according to the dose of vitamin B complex (20 mice in each group): V1 group (injected with 150μL normal salin), V2 group (injected with 50μL vitamin B complex solution+100μL normal saline), V3 group (injected with 100μL vitamin B complex solution+50μL normal saline), and V4 group (injected with 150μL vitamin B complex solution). After 4 weeks of treatment, the vaginal secretion was obtained for microscopic smear examination. HE stainning was performed to observe histopathological changes of vaginal tissues. The expressions of inflammatory factors were detected by ELISA. Meanwhile, VVC model of vaginal epithelial cells was established. The effects of different concentrations of vitamin B complex on anti-fungal effect of fluconazole were detected in vitro. After the treatment, complicated patients in the group C had significantly higher effective rates than those in the group A and group B. After the intra-gastric administration, the microscopic smear examination found that obvious pseudohypha in cluster with a lot of blastospores can be seen in the vaginal secretions of mice in the V1 group under the microscope. There was significant difference between mice treated with different dosages of vitamin B complex. The inflammatory response of mice in the V1 group was significantly higher than those in other groups and the inflammation response reduced with the increase of vitamin B complex dosage. The vitamin B complex elevated the curative effects of fluconazole on VVC model of vaginal epithelial cells and significantly increased the anti-fungal effect of fluconazole. Our findings suggest that vitamin B complex could be an effective adjuvant therapy for complicated VVC.

Relationship of HPV infection and BV, VVC, TV: a clinical study based on 1 261 cases of gynecologic outpatients

Objective: To Explore the relationship between HPV infection and bacterial vaginosis(BV), vulvovaginal candidiasis(VVC), and trichomonal vaginitis(TV). Methods: Clinical data from 1 261 gynecologic outpatients who underwent the vaginal microecology and HPV type detection during June 2015 to December 2015 were collected and analyzed in the First Affiliated Hospital of Xi'an Jiaotong University. Results: In 1 261 patients, 328 cases infected with HPV, infection rate was 26.01%(328/1 261); vaginal infectious disease in 328 cases of HPV infections were 219 cases(66.8%, 219/328), vaginal infectious diseases in 933 cases of HPV uninfected were 503 cases(53.9%, 503/933), incidence of vaginal infectious disease in HPV infected patients was higher than that in HPV uninfected patients(χ2=5.87, P=0.01). 142 cases of BV had 54 cases infected with HPV(38.0%, 54/142), 296 cases of intermediate type BV had 88 cases infected with HPV(29.7%, 88/296), 231 cases of normal vaginal microecology had 51 cases infected with HPV(22.1%, 51/231), 99 cases of VVC had 15 cases infected with HPV(15.2%, 15/99), 2 patients with TV had 0 cases infected with HPV(0/2), HPV infection rate in BV, intermediate type BV patients were significantly higher than normal patients(P<0.05), while there were no statistical differences among VVC, TV and normal patients(P>0.05). The intensity of HPV infection were positively correlated with BV, intermediate type BV(OR=2.17, 95% CI: 1.37-3.43, P<0.01; OR=1.49, 95% CI: 1.00-2.22, P= 0.04); while, VVC, TV were uncorrelated with HPV infection(all P>0.05). Conclusions: BV, intermediate type BV are positively correlated with HPV infection, especially for the high-risk HPV. VVC and TV are not correlated with HPV infection.

Identification of Candida species Isolated From Vulvovaginal Candidiasis Patients by Chromgen agar and PCR-RFLP Method

This study focuses on diagnosis of Candida species causing Vulvovaginal Candidiasis using phenotype and genotype analyzing methods, and frequencies of candida species also using Vulvovaginal Candidiasis patients. 130 samples (100 from patients and 30 from non infected women) were collected and cultured on biological media. Identifying the yeasts, initially some phenotypic experiments were carried out such as germ tube, from motion of pseudohyphae and clamydospores in CMA+TW80 medium, API20 candida and CHROMagar Candida. Genomic DNA of all species were extracted and analyzed with PCR and subsequent Polymerase Chain Reaction - Restriction Fragments Length Polymorphism (PCR-RFLP) methods. Frequency of C. albicans, C. krusei, C. tropicalis , C. parapsilosis and C. glabrata were 46.4%, 31%, 18%, 7.2%, and 1.8%, respectively.The ITS1-ITS4 region was amplified and the Restriction enzyme Msp1 digests this region and was used to identify of candida species .Electrophoretically ribosomal DNA of C. albicans, C. krusei, C. tropicalis and C. glabrata produced two bands whereas the C. parapsilosis gave one band.