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 nonalbicans 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

[Comparision of antifungal effect of Nystatin alone and in combination with nanosilver particles against candida species isolated from chronic candidal vaginitis]

Nystatin is a polyene with antifungal effect which is used in treatment of cutaneous and mucocutaneous candidiasis in diverse forms. Increasing its utilization in recent years has led to predominant increasing of resistances. Nowadays for increasing of antifungal effect and decreasing of resistance and side effects of drugs they are used in combination with each other. So we decided to investigate antifungal effect of Nystatin in combination with nanosilver particles. This was an experimental study which has been accomplished on 30 samples of isolated candida species from patients effected to chronic candidal vaginitis. In this study the antifungal effects of Nystatin and silver nanoparticles each of them alone and in combination with each other by microdilution broth, were examined. Findings were described on the base of logestic regression and man-vitni exam. Findings suggested that Nystatin was able to inhibit the growth of candida species at an expanded range of concentration between 16-128 microgeram per milliliter. As well antifungal activity of Nystatin with silver nanoparticles was increased in comparision with using Nystatin alone. Introduce of nanosilver in drug formation of Nystatin can be useful in treatment of chronic vaginal candidiasis

[Investigation of prevalence of onychomycosis due to yeast fungi in dystrophic nails of patients who referred to Razi hospital [2010 - 2011]]

Yeasts are one of the most common causes of onychomycosis. In countries like Iran, Saudi Arabia, Italia and Spain, yeasts are reported as the most frequent causes of onychomycosis. Inhibition of the immune system such as chemotherapy, radiation therapy, the use of broad spectrum antibodies, aggressive treatment with corticosteroids, HIV and diabetes melitus predispose the body with these fungal infections. Onychomycosis due to yeast has a higher prevalence in finger nails and its incidence in women is two or three times more than men. In some occupations such as nurses, dish washers, confectionery and housewives it can be more observed. Among the yeasts, Candida albicans is the most common agent onychomycosis. This study has been designed for investigation of prevalence of onychomycosis due to yeast in patients who referred to Razi hospital. This was a cross sectional study and 700 dystrophic nail samples were examined by both direct examination and culturing. In direct examination KOH [hydroxide potassium] 20% and for culturing saboroud dexterose agar [S] media were used. For identification yeasts complementary examination were done such as: Reynold braude test, API test and culturing on candida chrom agar media and corn meal agar media were used. For investigation of relevance between variables, Chisquare test and Fisher exact test were used. Of 700 dystrophic nail samples [15.71%], 110 samples were yeast positive by both direct examination and culturing. Thirty one patients were males and 79 patients were females and in both sexes those most infected were between 50-59 years of age [27.3%]. Eighty patients had fungal infection of finger nails and 16 patients had fungal infection on toe nails; 14 patients had both infections on finger and toe nails. The most frequent detected yeast species was Candida albicans [42.7%] which was followed by Candida parapsilosis [20.9%], Candida tropicalis [14.5%], Candida krusei [12.7%], Candida glabrata [3.6]%, Candida gillermondi [2.7%], and Candida lousitani, famatata, rodotroula [each 0.9%] .The most common clinical type noted was distal subungual onychomycosis in 50% of cases. In this study 52 patients [48.1%] had diabetes which was the most common disease between patients with onychomycosis due to yeasts. Diagnosis of onychomycosis due to yeast is very important because it shows the person's immune response. Identification of pathogenic yeast species in terms of epidemiology and selecting appropriate and effective treatment is important

[Evaluation of Antifungal activity of new derivatives of Imidazole using Colorimetric method]

The incidence of invasive fungal infections has been increasing for two decades. A matter of concern in the treatment of fungal infections is the limited number of efficacious antifungal drugs. Many available drugs lead to the development of resistance, In order to seek new antifungal agents we assessed the antifungal activity of newly synthesized Imidazol compounds by a colorometric method. In this experimental study antifungal activity of the new Imidazol compounds against Candida albicans, Saccharomyces cervisiae, Aspergillus niger, Microsporum gypseom was investigated by colorometric method and results were compared to microdilution ones. 2-hydroxyphenacyl-azole and 2- hydroxyphenacyl-azolium compounds have been identified as a new class of azole antifungal agents with a good spectrum of activity. The colorimetric method is a simple microtiter method for determining the susceptibility of species of fungal against antifungal agents. Most derivatives showed significant in vitro antifungal activities against tested fungi with low MIC [minimum inhibitory concentration] values included in the range of 0.25-32 micro g/mL comparable to the reference drug Fluconazole

[Report of 34 cases of saprophytic fungi isolated from dystrophic nails of patients referred to Razi Hospital [2010 - 2011]]

Saprophytes are one of the agents causing nail dystrophy. Saprophytes can invade healthy nail or may invade nails previously damaged in the course of other diseases and grow with suitable conditions. The reported incidence of saprophytic nails is between 1.43-17.6%. Saprophytes preferably invade the nails on the big toes, especially in individuals above 60 years. The most etiologic agents of saprophyte nail are Aspergillus spp, Acremonium spp, Scopulariopsis spp, Penicillium spp, and fusarium. The purpose of this study was to determine the prevalence agents of saprophytic nails in patient that had referred to Razi hospital. This was a cross sectional study and nail samples were analyzed by direct microscopy and culture. Microscopic examination of these specimens was carried out in potassium hydroxide solution [20%]. These specimens were cultured on two media of sabourad dextrose Agar [S]. Czapek-Dox Agar [CZA] medium was used for identification of Aspergillus species. For investigation of relevance between the variables, Chi-square test and Fisher exact tests were used. In this study, 34 cases were positive by both direct microscopy and culture. Of those, 17 patients were females and 17 patients were males. The most frequently isolated saprophytes from nails was Aspergilus flavous [35.3%]. Meanwhile 58.8% of saprophytes were isolated from toe nails. In this study the distal subungual onychomycosis was the most frequent [% 64.7%]. The age group 50-59 years [29.4%] had the highest prevalence of saprophytic nail infections.In this study the prevalence of saprophytic nails infections was 17.2%. A proper diagnosis, consisting of both clinical and mycological examinations, may aid the clinician in selecting the most appropriate therapy. Knowledge of epidemiology and mycology characteristics of nail infections has been noted by many authors as being an important tool for control of these fungal infections

[Comparision of antifungal effect of fluconazole alone and in combination with nanosilver particles against candida species isolated from chronic candidal vulvovaginitis]

Candidal vulvovaginitis is a female genital infection that is occured by the over growth of candida species and specially Candida - albicans and occasionally it appears as recurrent and chronic and resist to therapy. Fluconazole is one of the current drugs that is used in treatment of this disease and sometimes resistance is observed to this. Therefore indirection of making the therapy better we decided to investigate the activitiy of fluconazole in combination with silver nanoparticles against candida species isolated from chronic and recurrent candidal vaginitis. This was an experimental study with convenient sampling that was performed on 30 patients. All specimens were examined in direct microscopy, culturingand differential tests to identify different candida species from each other such as culture on candida chrom agar, germ tube, temperature test and sugar assimilation with API. Then the antifungal effects of fluconazole and silver nanoparticles, each of them alone and in combination with each other, were examined .Findings were described on the base of logestic regression and man-vitni exam. In our study 30 specimens of chronic genital candidiasis were diagnosed with isolated agents Candida albicans, glabrata, krusei, tropicalis, parapsilosis and Candida famata respectively. Also findings suggested that fluconazole was able to inhibit the growth of candida species at an expanded range of concentration between 4-128 microgeram per milliliter.As well antifungal activity of Fluconazole with silver nanoparticles was increased in comparision with using Fluconazole alone. For the prevention of recurrent cases and to stabilish correct diagnosis it is essential to carry out sensitivity and diagnostic tests in laboratory and also administration of silver nanoparticle in combination with fluconazole in drug formulation for topical uses in treatment of chronic vaginal candidiasis and inhibition of recurrent cases, can useful

Anticandidal effects of shallot extracts against chronic candidiasis agents

Shallots are an important part of the diet of many population and there is long-held belief in their health enhancing properties. At least one episode of vaginal candidiasis is reported in up to 75 percent of women that Candida albicans is responsible for 80 to 92 percent of episodes of vaginal candidiasis in the world. The aim of this study was to determine anticandidal activity of shallot against chronic candidiasis agents. This study is an experimental study. With use of microdilution method, antifungal activities of alcoholic and aqueous extracts of shallot [allium hirtifolium] were tested against 33 Candida species in vitro that isolated from patient with chronic candidiasis that referred to Mirza Koochak khan and Lolagar hospitals. The results showed antifungal activity of Allium hirtifolium against all the Candida species tested and anticandidal activity of the alcoholic extract was much better than aqueous one. The results indicated that crude juice of shallot has anticandidal activity and might be promising in treatment of candidiasis

[Evaluation of serological diagnosis indirect immunofluorescence and ELISA in comparison to direct examination and culture in vulvovaginal candidiasis]

Vulvovaginal candidiasis occurs due to the overgrowth of candida in genital system mucosa of females. Symptoms and signs of vulvovaginal candidiasis are unspecific, therefore diagnosis is not certain. The aim of this research was comparison of the result of indirect immunofluorescence and ELISA with culture and direct microscopy examination in patients with vulvovaginal candidiasis. This was a comparative-descriptive study performed on 87 patients and 50 normal cases as controls. All specimens were examined using direct microscopy, culturing and complimentary test to differentiate the candida species from each other. Serological tests such as indirect immunofluorescence and ELISA were performed on sera of the patients. To compare the quantitative and qualitative data, t-student test, chi-square and exact fisher test were used, if necessary. Out of 87 specimens, 50 cases were diagnosed as vulvovaginal candidiasis. The most frequent isolated pathogens were C.albicans, C.glabrata, C.kefyr, C.inconspicua and C.famata, respectively. Also, in control group, the most frequent pathogens were C.albicans, C.glabrata and C.kefyr, respectively. In this study all of normal cases were negative in indirect immunofluorescence test and in patients group 42 person [84%] were positive and 8 [16%] negative. Control participants were negative in ELISA and in patient group 40 person [80%] were positive and 10 person [20%] negative. It seems in cases that direct microscopic and culturing methods is impossible, ELISA and indirect immunofluorescence can be used as an alternative method

[ comparison of the bayer acryl and acropars acryl effect on the adhesion of candida albicans]

Candidiasis is one of the most common fungal infections in oral cavity. About 85% of this infection is caused by Candida albicans a flora of mouth. Since one of the problems in denture users is it's contamination with C.albicans and discoloration, and lack of information about adhesion of C.albicans to different resins, we designed this study to compare the effect of two types of acrylic resin on adhesion of C.albicans. This was an experimental study within that 36 samples from Bayer and Acropars acrylic resin were added in tubes containing suspension of 1-106 [CFU/ml] of C.albicans. Then tubes were incubated at 37? c for 40 and 120 minutes. After incubation, the samples were transferred in 1ml of sterile saline and after resuspension by shaking a known quantity of each was inoculated on sabouraud dextrose agar and incubated at 37? c for 48 hours and after that, formed colonies were counted and were analyzed by Mann-u-Whitney test. These two acrylic resins did not reveal statistical significant differences in two 40? c and 120? c different lengths of the time from the point of adhesion. Also Bayer acrylic resin did not show any significant differences from the point of adhesion in to different lengths of times. But dissimilarly was significant for Acropars acrylic resin which indicated adhesion with the passage of the time. [p<0.1] These two Acrylic resins do not have any priority to each other from the view point of non-adhesion to Candida albicans, but Bayer acryl is more appropriate particularly for those who are with weak hygiene and are more susceptible to fungal infections. Surface roughness of acrylic resins and their components, characteristic and surface factors have essential role in adhesion of C.albicans

[Investigation of serum C-reactive protein and mannose binding lectin levels in patients with dermatophytosis]

Dermatophytosis is common cutaneous fungal disease with worldwide distribution. Interleukin8 [IL-8] realized from keratinocytes in the presence of dermatophytic antigens causes induction of acute responses in dermatophyte infection and subsequently production of acute phase proteins occurs in hepatocytes. C-reactive protein [CRP] and Mannose binding lectin [MBL] are acute phase proteins. Since few researches in the case of acute phase proteins in dermatophytic infections has been accomplished, this study has been designed for determining serum CRP and MBL levels in patients affected to dermatophytosis. This was a cross sectional study and samples were carried out on 96 healthy individuals and 105 patients affected to dermatophytosis with non probable and in access procedure. For isolation and identification of dermatophyte direct microscopic examination, culturing and complementary examinations were done and for determination of serum CRP and MBL levels in healthy individuals and in patients ELISA test were used. For investigation of relevance between variables, Chi-square, Fisher exact, Mann-Whitney and Roc curve analysis were used and p<0.05 was considered as meaningful level. The median serum CRP level in healthy individuals and in patients group was 3.31 +/- 3.32 micro g/ml and 16.60 +/- 35.96 micro g/ml [p<0.001] respectively and the median serum MBL level was 1.53 +/- 1.87 micro g/ml and 1.97 +/- 2.03 micro g/ml [p=0.039] respectively. CRP [p<0.001] and MBL [p=0.042] were determined meaningful parameters for dermatophytosis. MBL deficiency [MBL concentrations<1 micro g/ml] was higher in control subjects [56.2%] than in patients [41.0%]. Findings of this study indicate increased concentrations of CRP and MBL in patients affected to dermatophytosis and their role in this infection. Probably observation of high frequency of MBL deficiency in healthy individuals in compare with patients group indicates that it is not predisposing factor in affecting to dermatophytosis