Effect of pepstatin A on the virulence factors of Candida albicans strains isolated from vaginal environment of patients in three different clinical conditions

The aspartate proteinase inhibitor pepstatin A was used to study a possible correlation among proteinase activity and other virulence factors of Candida albicans strains isolated from the vaginal environment of patients in three different clinical conditions: asympthomatic, vulvovaginal candidiasis (VVC) and recurrent vulvovaginal candidiasis (RVVC). The addition of 1.0 μM pepstatin A did not have any significant effect on hyphae formation, biofilm production and in the cell surface hydrofobicity of isolates in the three different clinical conditions. However, pepstatin A reduced the adherence of C. albicans to vaginal mucosa epithelial cells (53.1, 48.7 and 59.9%, respectively to isolates from asymptomatic, VVC and RVVC patients). This result suggests that the secreted aspartate proteinases (Saps) of this fungal pathogen may have auxiliary roles in cellular adhesion.     

Vulvovaginal candidiasis is associated with the production of germ tubes by <Emphasis Type="Italic">Candida albicans</Emphasis>

Twenty Candida albicans strains isolated from women attended at the Teaching and Research in the Laboratory of Teaching and Research in Clinical Analysis of the State University of Maringa, Paraná, Brazil, have been analyzed. Yeasts were identified by classical methods and patients subdivided into asymptomatic, vulvovaginal candidiasis(VVC) and recurrent vulvovaginal candidiasis (RVVC) groups. Yeasts were incubated in RPMI + fetal calf serum to analyze germ tubes every two hours, up to 10 h. In vitro sensitivity to fluconazole, itraconazole, ketoconazole, amphotericin B and nystatin was analyzed according to NCCLS-M27-A microdilution assay. Yeast isolated from symptomatic women produced significantly more germ tubes than asymptomatic women (P < 0.05). However, no significant difference between yeasts from VVC and RVVC occurred (P > 0.05). Variation between MIC₅₀ and MIC₉₀ of tested antifungal agents was slight among isolated yeasts, while no resistant yeasts were detected. Nevertheless, VVC yeasts were more DDS (reduced dose-dependent susceptibility) for nystatin and RVVC were more DDS for ketoconazole. Results suggest that colonization by yeast in the vagina and lack of symptoms may be partially explained by the yeast’s sparse capacity to form germ tubes, On the other hand, RVVC was not associated with antimicrobial resistance. DDS high frequency for nystatin and ketoconazole indicates that identification, and susceptibility of antifungals tests are important to management of VVC.     

A New Model of Vaginal Infection by <Emphasis Type="Italic">Candida albicans</Emphasis> in Rats

Vulvovaginal candidiasis (VVC) is regarded as an important public health issue, and several aspects of its pathogenesis are not yet sufficiently clear. Experimental in vivo models of vaginal infection with Candida albicans have been extremely useful in the identification of factors concerning hormonal influences on the infection, the virulence of the yeasts, the susceptibility, and the treatment of the infection. The development of easily manageable, reproducible, and economically viable animal models of VVC is highly important. We describe a simple experimental model of VVC in rats, using a pharmaceutical brand of estradiol hexa-hydrobenzoate for human treatment. All the steps of this model were standardized; and after the experiments, the rats were euthanized for further examination of their tissues by scanning and transmission electron microscopy. Standardized features included the use of non-ovariectomized rats, sterile distilled water as the hormone vehicle, estradiol hexa-hydrobenzoate administered at 0.20 mg/week/rat fractionated three times/week, and a yeast suspension of 5 × 10⁸ yeasts/ml in a single vaginal administration 1 week after hormone induction. In this way, 100% of the rats were in pseudo-estrus and developed and maintained the infection until the third week of the experiment. Electron microscopy observation of the vagina of the rats confirmed the presence of both pseudo-estrus and vaginal infection. The standardized experimental model proved inexpensive, reproducible, and easily feasible for the induction of vaginal infection with C. albicans and may help to clarify important aspects of the pathogenesis and treatment of VVC.     

Vaginal microbiota in healthy pregnant women and prenatal screening of group B streptococci (GBS)

The microbiota of the lower female genital tract was evaluated from vaginal swabs obtained from 623 healthy pregnant women at gestation periods of 35–40 weeks. Isolated and identified microorganisms were expressed as percentages of total samples. As expected, lactobacilli made up the dominant vaginal microbiota (70%). Enterobacteriaceae, mainly Escherichia coli, Klebsiella spp. and Proteus, were present in 38% of the samples, which might reflect the possible contamination of vaginal tract with rectal microorganisms. Candida albicans was present in 10% of healthy pregnant woman assayed. Streptoccocci (Streptococcus sp. and Enterococcus faecalis with 3% and 4%, respectively) and other gram-positive cocci (Staphylococcus sp., 5%), along with other microorgansisms such as Gardnerella vaginalis (5%) and Pseudomonas aeruginosa (2%) may represent a potential infection risk. Streptococcus agalactiae (group B streptococci β-hemolytic, GBS) was detected in 7% of the samples. GBS infection is a leading cause of neonatal morbidity and mortality in the developed world. Furthermore, GBS was often co-isolated with C. albicans (54.5%) in the samples. A complete and detailed evaluation of the vaginal biota swab, with particular attention to the presence of potential pathogens such as GBS, is a preventive strategy that can provide useful information to obstetricians and gynecologist in managing the last days of pregnancy and delivery. Electronic Publication     

The Role of 17β-Estrogen in Candida albicans Adhesion on Human Vaginal Epithelial Cells via FAK Phosphorylation

Purposes

To investigate the role of 17β-estrogen in Candida albicans (C. albicans) adhesion on human vaginal epithelial cells in vulvovaginal candidiasis (VVC).

Methods

The vaginal epithelial cell line, VK2/E6E7, was used to study the estrogen-induced molecular events between C. albicans and cells. An adhesion study was performed to evaluate the involvement of the estrogen-dependent focal adhesion kinase (FAK) activation in cell adhesion. The phosphorylation status of FAK and estrogen receptor α (ERα) upon estrogen challenge was assessed by western blotting. Specific inhibitors for ERα were used to validate the involvement of ERα–FAK signaling cascade.

Results

A transient activation of ERα and FAK was observed following the stimulation with 1000 nM estrogen for 48 h, as well as the increased average number of C. albicans adhering to each vaginal epithelial cell. Estrogen-induced activation of ERa and FAK was inhibited by the specific inhibitor of ERα, especially when the inhibitor reached a 10 μM concentration and allowed to act for 12 h. Simultaneously, a decrease in the number of adherent C. albicans was observed. However, this inhibitory effect diminished as the concentration of estrogen increased.

Conclusion

FAK and ERα signaling cascades were involved in the early interaction between the vaginal epithelial cells and C. albicans, which appeared to be linked with the enhanced cell adhesion leading to VVC and promoted by a certain concentration of estrogen.

     

Genotyping and Persistence of <Emphasis Type="Italic">Candida albicans</Emphasis> from Pregnant Women with Vulvovaginal Candidiasis

Objective

To study Candida albicans genotypes using RAPD and their susceptibility to fluconazole in healthy pregnant women and in vulvovaginal candidiasis (VVC) patients after topical treatment with clotrimazole.

Methods

Vaginal swabs were collected at t = 0 and t = 1 (1 month later) in pregnant women (control group, n = 33), and before (t = 0), at 1 month (t = 1) and at 2 months (t = 2) after clotrimazole treatment in pregnant women with VVC.

Results

Candida albicans was isolated in 30% of healthy pregnant women and 80% of patients with VVC. A high genetic heterogeneity was observed in C. albicans genotypes between individuals. In patients with VVC, topical antifungal treatment with clotrimazole was clinically effective, but only in a 62% C. albicans was eradicated. In patients in which C. albicans was not eradicated, this microorganism persisted for 1 or 2 months after the antifungal treatment. The persistent colonies were not associated with a specific genotype, but they were associated with higher MICs in comparison with colonies isolated from the control group.

Conclusions

Therapy with topical clotrimazole, despite a good clinical outcome, could not eradicate completely C. albicans allowing the persistence of genotypes, with higher MICs to fluconazole. More studies with higher number of patients are needed to validate this preliminary finding.

     

IL-22 and IDO1 Affect Immunity and Tolerance to Murine and Human Vaginal Candidiasis

The ability to tolerate Candida albicans, a human commensal of the gastrointestinal tract and vagina, implicates that host defense mechanisms of resistance and tolerance cooperate to limit fungal burden and inflammation at the different body sites. We evaluated resistance and tolerance to the fungus in experimental and human vulvovaginal candidiasis (VVC) as well as in recurrent VVC (RVVC). Resistance and tolerance mechanisms were both activated in murine VVC, involving IL-22 and IL-10-producing regulatory T cells, respectively, with a major contribution by the enzyme indoleamine 2,3-dioxygenase 1 (IDO1). IDO1 was responsible for the production of tolerogenic kynurenines, such that replacement therapy with kynurenines restored immunoprotection to VVC. In humans, two functional genetic variants in IL22 and IDO1 genes were found to be associated with heightened resistance to RVVC, and they correlated with increased local expression of IL-22, IDO1 and kynurenines. Thus, IL-22 and IDO1 are crucial in balancing resistance with tolerance to Candida, their deficiencies are risk factors for RVVC, and targeting tolerance via therapeutic kynurenines may benefit patients with RVVC.     

The liquid Kangfuxin (KFX) has efficient antifungal activity and can be used in the treatment of vulvovaginal candidiasis in mice

Vulvovaginal candidiasis (VVC) is an infectious disease caused mainly by Candida albicans. Kangfuxin (KFX) is a traditional Chinese medicine preparation made from Periplaneta americana extracts, which promotes wound healing and enhances body immunity and also acts as an antifungal agent. Here, we evaluated the effect of KFX in the treatment of VVC in vitro and in vivo. The minimum inhibitory concentration (MIC₅₀) of KFX against C. albicans ranged from 7·65 to 20·57%. In addition, KFX was more efficient than fluconazole (FLC) in inhibiting the drug-resistant C. albicans, and the effect was more intense after 8 h. The KFX treatment also exhibited good activity in vivo. It restored the body weight and reduced the vulvovaginal symptoms in mice induced with VVC. It downregulated the expression of the hyphae-related gene, HWP1, thus inhibiting the growth and development of C. albicans hyphae. It also increased the number of neutrophils and promoted the secretion of interleukin-17A (IL-17A); however, the levels of interleukin-8 (IL-8) and interleukin-1β (IL-1β) decreased in mice with VVC. We deduce that KFX effectively treats vaginal candidiasis in two ways: by inhibiting the growth and development of mycelia to reduce colonization of C. albicans and by promoting the secretion and release of IL-17A and neutrophils in high numbers to fight C. albicans infection. This study provides a theoretical basis for the use of KFX for the clinical treatment of VVC.     

Vertical Transmission of <Emphasis Type="Italic">Chlamydia trachomatis</Emphasis> in Chongqing China

This is the first study to investigate vertical transmission of Chlamydia trachomatis in Chongqing China. For this study, 300 cervical swab samples from pregnant women and 305 nasopharygeal swab samples from their babies (605 specimens) were collected for nest polymerase chain reaction (nPCR) of the ompl gene, which encodes the major outer membrane protein (MOMP) and typed C. trachomatis using Cleavase fragment-length polymorphism (CFLP) labeled with digoxin. From these samples, 11% (33/300) of pregnant women samples were successfully amplified. The vertical transmission rate of C. trachomatis from mother to baby was 24% (8/33). The vertical transmission rates were 66.7% (6/9) for mothers with vaginal delivery and 8.3% (2/24) for those with cesarean section. The incidence of premature membrane rupture among C. trachomatis-positive pregnant women was 30.3% (10/33), which was greater than among those who were C. trachomatis-negative (13.5%, 36/267; χ² = 4.2; p < 0.05). Four genotypes including type E (3 pairs), type F (2 pairs), type H (2 pairs), and type D (1 pair) were observed by CFLP assay labeled with digoxin and confirmed by DNA sequencing in the 16 C. trachomatis-positive samples from eight pregnant women and their eight infants. Each pair of matched maternal–infantile samples showed identical CFLP. This study showed the incidence of C. trachomatis infection in pregnant women, the vertical transmission rate for C. trachomatis, and the genotypes of C. trachomatis in Chongqing, China. The CFLP assay labeled at the 5′ end of the forward primer with digoxin was first used successfully to genotype of C. trachomatis. As a promising method for C. trachomatis genotyping, CFLP had good sensitivity, reproducibility, and simplicity and no radioactive contamination.     

复发性外阴阴道念珠菌病的致病菌种鉴定及药敏分析

目的:探究复发性外阴阴道念珠菌病致病菌的菌种分布及耐药情况,指导临床治疗。方法:收集210例念珠菌性外阴阴道炎患者阴道分泌物标本,其中复发性外阴阴道念珠菌病(RVVC)组76例,外阴阴道念珠菌病(VVC)组134例。运用科玛嘉显色培养基法进行菌种鉴定和药物敏感试验。结果:在210例病例中,RVVC组中白色念珠菌54株(71.05%),光滑念珠菌15株(19.74%),热带念珠菌4株(5.26%),克柔念珠菌2株(2.63%),近平滑念珠菌1株(1.32%)。非白色念珠菌中以光滑念珠菌为主,在RVVC组中的比例明显高于VVC组,有显著性差异(P<0.05)。药敏试验显示,RVVC的念珠菌株中仅8株对7种药物全部敏感,23株敏感药物<5种,其中1株仅对制霉菌素敏感;对药物的敏感率为制霉菌素(97.37%)>克霉唑(60.52%)>酮康唑(51.32%)>伊曲康唑(36.84%)>咪康唑(35.53%)>氟康唑(23.68%)>特比奈芬(10.53%)。结论:白色念珠菌仍是RVVC的主要致病菌,非白色念珠菌比例在RVVC组中显著高于VVC组,其中以光滑念珠菌为主。念珠菌对制霉菌素敏感率最高,对唑类药物耐药性有增加趋势。     

复发性外阴阴道念珠菌病的致病菌种鉴定及药敏分析

目的：探究复发性外阴阴道念珠菌病致病菌的菌种分布及耐药情况,指导临床治疗。方法：收集210例念珠菌性外阴阴道炎患者阴道分泌物标本,其中复发性外阴阴道念珠菌病（RVVC）组76例,外阴阴道念珠菌病（VVC）组134例。运用科玛嘉显色培养基法进行菌种鉴定和药物敏感试验。结果：在210例病例中,RVVC组中白色念珠菌54株（71.05%）,光滑念珠菌15株（19.74%）,热带念珠菌4株（5.26%）,克柔念珠菌2株（2.63%）,近平滑念珠菌1株（1.32%）。非白色念珠菌中以光滑念珠菌为主,在RVVC组中的比例明显高于VVC组,有显著性差异（P〈0.05）。药敏试验显示,RVVC的念珠菌株中仅8株对7种药物全部敏感,23株敏感药物〈5种,其中1株仅对制霉菌素敏感;对药物的敏感率为制霉菌素（97.37%）〉克霉唑（60.52%）〉酮康唑（51.32%）〉伊曲康唑（36.84%）〉咪康唑（35.53%）〉氟康唑（23.68%）〉特比奈芬（10.53%）。结论：白色念珠菌仍是RVVC的主要致病菌,非白色念珠菌比例在RVVC组中显著高于VVC组,其中以光滑念珠菌为主。念珠菌对制霉菌素敏感率最高,对唑类药物耐药性有增加趋势。     

Efficacy of CIM 1166, a combination of compounds derived from Mentha spp. in alleviating experimental vulvovaginal candidiasis in mice

Candida albicans is yeast that is most often associated with serious fungal infections and can cause fungal diseases in immuno-compromised patients especially patients suffering from AIDS, cancer and cases of organ transplant. Amongst women, candidal vaginitis is predominantly caused by strains of Candida albicans and also remains to be a common problem in immuno-competent or healthy women. A study was undertaken to assess the efficacy of a compound CIM 1166 obtained from plant source which was found to possess promising antimicrobial property under in vitro conditions especially against C. albicans. Taking the lead further, a small animal model utilizing aged Swiss albino females that had parturated at least three times were taken up for model development. Infection (7 × 10⁶ cfu/ml) was instilled into the vagina in a volume of 20 μl for 3 days. Vaginal washings were aseptically collected on day 4th to confirm the establishment of infection following which the treatment was started which continued for the next 5 days through vaginal route. Vaginal washings were collected on 6th day and the colony forming units were enumerated on chloramphenicol incorporated SDA plates. The results indicated that there was a significant decrease in the colony forming units in vaginal washings (8.0 × 10² cfu/ml) of the treated animals as compared to blank control group (6.0 × 10⁴ cfu/ml). The positive control group administered with clotrimazole also showed a recovery from infection with a fungal load of 8.78 × 10² cfu/ml. The study proves the efficacy of CIM 1166 in curing vaginal candidiasis in mice, which can be taken up for formulation development and further studies.     

β-Glucan Induces Reactive Oxygen Species Production in Human Neutrophils to Improve the Killing of Candida albicans and Candida glabrata Isolates from Vulvovaginal Candidiasis

Vulvovaginal candidiasis (VVC) is among the most prevalent vaginal diseases. Candida albicans is still the most prevalent species associated with this pathology, however, the prevalence of other Candida species, such as C. glabrata, is increasing. The pathogenesis of these infections has been intensely studied, nevertheless, no consensus has been reached on the pathogenicity of VVC. In addition, inappropriate treatment or the presence of resistant strains can lead to RVVC (vulvovaginal candidiasis recurrent). Immunomodulation therapy studies have become increasingly promising, including with the β-glucans. Thus, in the present study, we evaluated microbicidal activity, phagocytosis, intracellular oxidant species production, oxygen consumption, myeloperoxidase (MPO) activity, and the release of tumor necrosis factor α (TNF-α), interleukin-8 (IL-8), IL-1β, and IL-1Ra in neutrophils previously treated or not with β-glucan. In all of the assays, human neutrophils were challenged with C. albicans and C. glabrata isolated from vulvovaginal candidiasis. β-glucan significantly increased oxidant species production, suggesting that β-glucan may be an efficient immunomodulator that triggers an increase in the microbicidal response of neutrophils for both of the species isolated from vulvovaginal candidiasis. The effects of β-glucan appeared to be mainly related to the activation of reactive oxygen species and modulation of cytokine release.     

外阴阴道念珠菌病、复发性外阴阴道念珠菌病及健康人群甘露糖结合凝集素水平的分析研究

目的观察健康人群、外阴阴道念珠菌病(VVC)和复发性外阴阴道念珠菌病(RVVC)的甘露糖结合凝集素水平。方法收集健康人群、VVC和RVVC病例,给予规范化治疗,观察治疗前、后各组MBL水平的变化,并进行随访。结果发现各组的MBL水平VVC组>RVVC组>健康组,但各组之间及各组本身治疗前后的MBL差异无显著性意义(P>0.05)。治疗后VVC组复发者5例,占14%,RVVC组复发者13例,达到48%,两组复发者与未复发者的MBL水平,经t检验无显著性差异(P>0.05)。结论 MBL的水平在VVC患者中的水平高于RVVC患者,两者的复发率基本与既往文献报道一致,对于VVC患者来说,MBL水平高于0.8 ng/mL似乎患RVVC的风险稍小些。     

Pulmonary Aspergillosis and Aflatoxins in Chronic Lung Diseases

Recent studies have clearly defined the vaginopathic Candida albicans strains that cause severe vulvovaginal candidiasis (VVC). Therefore, genotyping C. albicans isolates may predict the success of and assist in choosing the appropriate antifungal therapy. The purpose of this study was to compare the genotypes of C. albicans isolates causing VVC with those found in asymptomatic healthy pregnant and non-pregnant women in Adana, Turkey, as well as the antifungal susceptibility profiles of these isolates. A total of 216 independent C. albicans isolates were genotyped by allelic combination based on the microsatellite marker analysis of one such microsatellite, present in the promoter region of the elongation factor 3-encoding gene (CEF3) of C. albicans. The susceptibility testing profiles of all of the isolates against five antifungals and boric acid were obtained retrospectively from our laboratory records. We identified 20 genotypes on the basis of different allelic combinations at the CEF3 locus with a discriminatory power of 0.85. Genotypes 136–144 and 126–135 were present in 50 % of the isolates. No differences existed in the genotypic profiles of fungal isolates between pregnant and non-pregnant women. Remarkably, we did not find a single vaginopathic genotype. All of the isolates were susceptible to amphotericin B and 5-fluorocytosine, and the fluconazole and ketoconazole resistance rates were 0.9 and 3.7 %, respectively. Therefore, we did not find any correlation between genotype, severity of VVC, and antifungal resistance (P > 0.05). Even so, additional molecular data may provide new insights into the management of VVC.     

The Acute Neutrophil Response Mediated by S100 Alarmins during Vaginal Candida Infections Is Independent of the Th17-Pathway

Vulvovaginal candidiasis (VVC) caused by Candida albicans affects a significant number of women during their reproductive ages. Clinical observations revealed that a robust vaginal polymorphonuclear neutrophil (PMN) migration occurs in susceptible women, promoting pathological inflammation without affecting fungal burden. Evidence to date in the mouse model suggests that a similar acute PMN migration into the vagina is mediated by chemotactic S100A8 and S100A9 alarmins produced by vaginal epithelial cells in response to Candida. Based on the putative role for the Th17 response in mucosal candidiasis as well as S100 alarmin induction, this study aimed to determine whether the Th17 pathway plays a role in the S100 alarmin-mediated acute inflammation during VVC using the experimental mouse model. For this, IL-23p19^−/−, IL-17RA^−/− and IL-22^−/− mice were intravaginally inoculated with Candida, and vaginal lavage fluids were evaluated for fungal burden, PMN infiltration, the presence of S100 alarmins and inflammatory cytokines and chemokines. Compared to wild-type mice, the cytokine-deficient mice showed comparative levels of vaginal fungal burden and PMN infiltration following inoculation. Likewise, inoculated mice of all strains with substantial PMN infiltration exhibited elevated levels of vaginal S100 alarmins in both vaginal epithelia and secretions in the vaginal lumen. Finally, cytokine analyses of vaginal lavage fluid from inoculated mice revealed equivalent expression profiles irrespective of the Th17 cytokine status or PMN response. These data suggest that the vaginal S100 alarmin response to Candida does not require the cells or cytokines of the Th17 lineage, and therefore, the immunopathogenic inflammatory response during VVC occurs independently of the Th17-pathway.     

Proteinase detection,DNA typing and antimycotic susceptibility of Candida isolates from Colombian women with vulvovaginal candidiasis

Forty non-pregnant Colombian women (ages 18-45) with vulvovaginal candidiasis diagnosis (VVC) were enrolled in a blinded study to compare the efficacy of Itraconazole (ITRA) 400 mg vs. Fluconazole (FLU) 150 mg. Sexual partners received similar therapy. Proteinase detection by the Staib method and minimal inhibitory concentration (MIC) for FLU and ITRA by Etest method were performed in all Candida isolates. Patients were followed one year to determine clinical evolution and recurrence of VVC (RVVC). The strain identity of the RVVC isolates was determined by contour-clamped homogeneous electric field (CHEF) gel electrophoresis karyotyping and restriction fragment length polymorphism (RFLP). Thirty patients (75%) had one or two episodes of VVC/year, 83% of these were due to Candida albicans, while ten patients (25%) developed RVVC (three or more episodes/year); seven of them were treated with FLU. Non-C. albicans Candida species were detected in five of 30 (17%) of the patients with VVC and in seven of ten (70%) patients with RVVC (p=0.003). Isolates from nineteen patients were proteinase positive. Proteinase production and type of treatment were not related to recurrence of VVC (p>0.05). DNA typing revealed that in this population RVVC might be due to the same strain, substrain shuffling or different strains and species.     

Antifungal Susceptibility Profile of Candida Albicans Isolated from Vulvovaginal Candidiasis in Xinjiang Province of China

We investigated the antifungal susceptibility profiles of 207 independent Candida albicans strains isolated from patients with vulvovaginal candidiasis (VVC) in Xinjiang Province of China. Using CLSI M27-A3 and M27-S4 guidelines, anidulafungin and micafungin were the most active drugs against C. albicans showing an MIC₅₀/MIC₉₀ corresponding to 0.016/0.0313 µg/mL, followed by caspofungin (0.25/0.25 µg/mL), posaconazole (0.125/0.5 µg/mL), ravuconazole (0.063/1 µg/mL), itraconazole (0.125/1 µg/mL), amphotericine B (0.5/1 µg/mL), isavuconazole (0.063/2 µg/mL), 5-flucytosine (1/2 µg/mL), voriconazole (0.125/4 µg/mL), and fluconazole (0.5/4 µg/mL). 96.1% (199)–100.0% (207) isolates were sensitive to the three echinocandins tested, amphotericine B and 5-flucytosine. The in vitro activity of triazoles against all isolates tested was variable; itraconazole and voriconazole had reduced the activity to almost half of the isolates (55.1% (114) and 51.2% (106) susceptible, respectively). Fluconazole was active against 76.3% (158) isolates tested. The new triazoles ravuconazole, isavuconazole and posaconazole showed good in vitro potency against 89.9% (186)–95.2% (197) of isolates with the geometric mean MIC (µg/mL) of 0.10, 0.12 and 0.14 µg/mL, respectively. In conclusion, our study indicates that for effective management of systemic candidiasis in Xinjiang Province of China, it is important to determine the susceptibility profiles of isolated C. albicans from patients with VVC.

     

复发性外阴阴道假丝酵母菌病菌种分型及药敏

目的对RVVC、VVC患者阴道分泌物进行培养、菌种分型及药物敏感试验,探讨RVVC发生的原因。方法用沙保弱琼脂培养基培养阴道分泌物,分离纯化菌株;VITEK 2全自动微生物分析仪鉴定菌种;同时进行药物敏感试验。结果共分离出101株假丝酵母菌,其中RVVC组45株,VVC组51株,健康组5株。101株中白色假丝酵母菌86株,占85.1%(86/101),RVVC组45株中35株为白色假丝酵母菌,占77.8%(35/45);VVC组51株中47株为白色假丝酵母菌,占92.2%(47/51),健康组5株中4株为白色假丝酵母菌,占80%(4/5)。RV-VC组非白色假丝酵母菌比例高,与VVC组比较差异有显著性(P<0.05)。RVVC组假丝酵母菌对唑类药物敏感性低于VVC组。结论VVC、RVVC的主要致病菌仍是白色假丝酵母菌,RVVC组非白色假丝酵母菌比例高于VVC组。RVVC组假丝酵母菌对氟康唑的敏感率明显低于VVC组。     

A polymerase chain reaction for detection of Brucella canis in vaginal swabs of naturally infected bitches

A PCR assay for the detection of Brucella canis in canine vaginal swab samples was evaluated, comparing its performance with that of bacterial isolation, serological tests, and a blood PCR assay. One hundred and forty-four female dogs were clinically examined to detect reproductive problems and they were tested by the rapid slide agglutination test, with and without 2-mercaptoethanol (2ME-RSAT and RSAT, respectively). In addition, microbiological culture and PCR were performed on blood and vaginal swab samples. The results of the vaginal swab PCR were compared to those of the other tests using the Kappa coefficient and McNemar test. Of the 144 females that were examined, 66 (45.8%) were RSAT positive, 23 (15.9%) were 2ME-RSAT positive, 49 (34.02%) were blood culture positive, 6 (4.1%) were vaginal swab culture positive, 54 (37.5%) were blood PCR positive, 52 (36.2%) were vaginal swab PCR positive, and 50.69% (73/144) were positive by the combined PCR. The PCR was able to detect as few as 3.8 fg of B. canis DNA experimentally diluted in 54 ng of canine DNA, extracted from vaginal swab samples of non-infected bitches. In addition, the PCR assay amplified B. canis genetic sequences from vaginal swab samples containing 1.0 × 10⁰ cfu/mL. In conclusion, vaginal swab PCR was a good candidate as a confirmatory test for brucellosis diagnosis in bitches suspected to be infected, especially those negative on blood culture or blood PCR; these animals may be important reservoirs of infection and could complicate attempts to eradicate the disease in confined populations.