Non-albicans Candida Isolates Research Articles

Copper IUD increases virulence of non-albicans Candida species isolated from women with vulvovaginal candidiasis

Objectives: The aim of the study was to investigate the effect of copper-bearing intrauterine device (IUD) use on the virulence of Candida species causing vulvovaginal candidiasis (VVC). The in vitro ability of Candida species to produce proteinase and phospholipase enzymes was studied, together with their antifungal susceptibility.Methods: Vaginal swabs from women with VVC were cultured and Candida species were identified. Participants comprised 132 women with culture-confirmed VVC, of whom 65 were using a copper-bearing IUD and 67 were not. Candida isolates were tested for their ability to produce proteinase and phospholipase as well as for their susceptibility to fluconazole and nystatin.Results: Proteinase production was higher in non-albicans Candida (NAC) isolates of IUD users compared with non-users (p = 0.017). IUD use was significantly associated with antifungal resistance of NAC isolates to fluconazole (p = 0.013) and nystatin (p = 0.018). By contrast, IUD use seemed to significantly reduce the production of proteinase by C. albicans (p = 0.041), with no effect on its antifungal susceptibility. There was a significant negative correlation between proteinase production in both C. albicans and NAC as well as sensitivity to fluconazole (r= −0.383, p < 0.05 and r= −0.497, p < 0.05, respectively).Conclusion: IUD use enhanced the virulence (proteinase production and antifungal resistance) of NAC but not C. albicans, indicating a variation in virulence between Candida species in response to IUD use. C. albicans responded better to fluconazole, whereas NAC isolates were more sensitive to nystatin.

In vitro antifungal activity and phytochemical characterization of Eugenia uniflora, Libidibia ferrea and Psidium guajava

The aim of this study was evaluate the in vitro antifungal activity of crude extracts from Eugenia uniflora, Libidibia ferrea and Psidium guajava. The extracts were obtained by turbo-extraction using water (AQ) or acetone-water (AC-W) (7:3, v/v) as solvents and lyophilized to obtain the crude extracts (CE). The CE were characterized by UV-Vis, TLC and HPLC. The activity of CEs was investigated against clinical isolates of Candida spp. and the Minimum Inhibitory Concentration (MIC), MIC50 and MIC90 were determinated. The analysis by TLC showed that all CEs presented polyphenols (flavonoids and tannins). The CEs from E. uniflora showed higher amount of polyphenols (30.35 ± 2.15%, AC-W) and the HPLC analysis revealed the tannins in all extracts. The CEs of E. uniflora showed MIC range from 1.9 to 500.0 µg/mL, and lower values of MIC50 and MIC90 against non-albicans Candida isolates. Regarding L. ferrea and P. guajava, the results showing MIC from 3.9 to 1000.0 µg/mL (CE-AQ) against C. albicans. The results demonstrate antifungal performance from CE against various species of Candida spp., suggesting that the herbal species may be use as new potential antifungal agents. Additionally, the polyphenol content can play a pivotal role in the antifungal properties of CE.

Fungal profile and antifungal susceptibility pattern of candidiasis in esophageal cancer patients.

This study was carried out to determine the fungal profile and antifungal susceptibility pattern in the brushing samples of candidiasis in patients with carcinoma of esophagus. The study was carried out in the Departments of Microbiology and Surgical Oncology of a regional cancer center from January 2017 to December 2017. Samples were collected under all aseptic precaution and Clinical Laboratory Standards Institute guidelines 2017 was followed for antifungal susceptibility testing. A total of 132 endoscopy brushing samples were collected from histological proven esophageal cancer patients and processed for fungal culture. Of which, 75 (56.81%) samples showed culture positivity and were recruited. Candida albicans in 40 (53.33%), Candida krusei in 25 (33.33%), Candida tropicalis in 7 (9.33%), and Candida glabrata in 3 (4%) patients were seen. Among the 40 C. albicans isolates, all were sensitive to caspofungin - 40 (100%), 34 (85%) showed sensitivity to fluconazole, and 32 (80%) showed sensitivity to flucytosine. C. krusei and C. tropicalis showed 100% sensitivity to caspofungin, and C. glabrata isolates showed 100% resistance to caspofungin and 80% resistance to Amphotericin B. The present study revealed the emergence of multidrug-resistant, nonalbicans Candida isolates in cancer esophagus patients with candidiasis in northeast India.

The emergence of non-albicans candidemia and evaluation of HiChrome Candida differential agar and VITEK2 YST\xae platform for differentiation of Candida bloodstream isolates in teaching hospital Kandy, Sri Lanka

BackgroundCandidemia is an emerging hospital-acquired bloodstream infection (BSI). It is common among severely ill and immunocompromised patients. Even following appropriate therapy in candidemia, recent studies reveal relative high mortality (40%). The global incidence of candidemia shows an incline. In Sri Lanka, candida speciation often difficult where basic facilities are less available. We have compared the risk factors, epidemiology, demography, and performance of HiChrome Candida differential agar (HiCA) characteristics with the VITEK2 YST platform for differentiation of Candida albicans (CA) and non-albicans candida (NAC) from blood culture isolate.MethodsThis is a laboratory-based cross-sectional study. Positive aerobic BACTEC blood cultures having yeast were identified using HiCA and VITEK2® platform. Epidemiology, risk factors, and clinical outcomes were compared between CA and NAC bloodstream isolates.ResultsOut of 120 positive yeast samples, VITEK2® has identified 110 (92%) as Candida sp. From that CA-34 (31%) and NAC-76 (69%) were isolated. Candidemia following NCA in neonates (p = 0.02), infants (p = 0.04) and adults (p = 0.02) in ICU and immunocompromised patients were significantly higher. Compared to CA, NAC bacteremia period prevalence (0.00041%) and incidence (0.23 per 100,000-person-years) was significantly high (p = 0.03). NAC 48 (63%) isolates were resistance to azoles. Exposure to antifungals (odds ratio (OR); p = 0.03), prolonged intensive care stay > 14 days (OR-3.3; p = 0.04), having a central venous line for > 8 days (OR-4.3; p = 0.03) and on immunosuppressive treatment (OR-2.4; p = 0.04) was significantly poses risk for NAC candidemia. Sen day mortality was significant among non-albicans cases (p = 0.03) while 30-day mortality was significant among albicans cases (p = 0.04). Compared to VITEK2®, the HiCA method was 93% sensitive and 93% specific in detecting CA.ConclusionCompared to CA, candidemia following NAC was high. NAC isolates were having a high percentage of fluconazole and voriconazole resistance. VITEK2 YST® platform provides antifungal susceptibility with minimal inhibitory concentration (MIC). Impact, this would highlight the use of rapid candida identification flat form with MIC to direct appropriate antifungals for candidemia. For that implementation of novel diagnostic facilities like the VITEK2 YST platform at a tertiary care facility is demanding.

Epidemiology and antifungal susceptibility testing of non-albicansCandida species colonizing mucosae of HIV-infected patients in Yaoundé (Cameroon)

Non-albicans Candida (NAC) species have emerged as potent pathogenic yeasts among HIV-infected patients. Authors evaluated the epidemiology and antifungal susceptibility testing of non-albicansCandida species colonizing Yaoundé (capital of the Republic of Cameroon, Central Africa) HIV-infected patients. The mucosal specimens were collected and submitted to the mycological diagnosis. Yeast isolates were identified by the Matrix Assisted Laser Desorption Ionisation - Time of Flight Mass Spectrometry (MALDI-TOF MS). The antifungal susceptibility testing was achieved by the CLSI-M27 protocols, and the interpretation of clinical break points (CBPs) and epidemiological cutoff values were in accordance with the CLSI-M60 and M59 recommendations. Four hundred and two patients were recruited and 1218 samples collected. The colonisation frequency was 24.1% and 304 yeasts isolated. Yeast isolates were 113 (37.2%) C. albicans, 2 (0.7%) C. africana and 172 (56.6%) NAC isolates. The NAC isolates were grouped into 13 species including C. krusei (18.1%), C. glabrata (10.9%), C. tropicalis (8.5%) and C. parapsilosis (5.9%) as the major ones. All the isolates appeared to be wild-type for amphotericin B and itraconazole. One (1/33) isolate of C. glabrata was resistant to fluconazole. C. arapsilosis isolates appeared all susceptible to fluconazole. C. tropicalis isolates presented 50% (13/26) resistance to fluconazole. The achieved results bring out new insights about epidemiology of NAC species in Cameroon. The results also highlight the resistance of NAC species to current antifungal drugs.

Characterization, Virulence Factors and Antifungal Susceptibility of Vulvovaginal Candida Isolated from Women at Qena, Egypt

ALTHOUGH the incidence of vaginitis caused by non-albicans Candida tends to be increased, C. albicans still the main causative agent of vaginitis Candida. Eighty-eight vaginal swab samples were collected from women with acute vaginitis in Qena, Egypt. Of 50 isolates, 39 admitted into C. albicans and 11 non-albicans Candida isolates (78% and 22% prevalence, respectively) were identified. Youths were more susceptible to infection with vulvovaginal Candida, the rate of infection decreased with increase education levels and the risk of infection was greater among douching use women. All isolates belonging to Candida taxa were positive to proteinase activity and 48 (96%) were lipase producers. Non-albicans Candida (C. glabrata, C. tropicalis and C. krusei) were more proteinase producers than C. albicans (P< 0.000). Compared with C. tropicalis, the other isolated Candida exhibited less lipase activity (P< 0.000). The higher lipase capacity of C. tropicalis may reflect their increased prevalence among non-albicans Candida group. Among five essential oils, cinnamon and clove oils showed strong efficacy against isolated Candida strains compared with miconazole antifungal.

In Vitro Antifungal Activity of a Medicinal Plant Extract Mixture Against Candida Species Isolated from Patients with Oral Stomatitis

Background: Ankaferd Blood Stopper® (ABS) is a hemostatic product comprising a standardized mixture of Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum, and Urtica dioica. It is used to control bleeding after extracorporeal injuries, traumatic cuts, dental operations, and surgical intervention. ABS was reported to exhibit antibacterial and germicidal activities. Objectives: This in vitro study aimed to determine the antifungal activity of ABS. Methods: In total, 114 Candida strains; 65 Candida albicans and 49 non-albicans isolated from the oral cavity of patients with oral stomatitis, as well as three reference strains of C. albicans ATCC 90028, C.parapsilosis ATCC 22019, and C.krusei ATCC 6258, were tested by agar well diffusion, disk diffusion, and time-kill curve methods in this study. The results obtained for ABS were compared with those obtained for amphotericin B (AMB). Results: ABS and AMB exhibited inhibitory zones with mean diameters of 18.2 ± 1.4 (12-20) mm, 20.6 ± 1.2 (18 - 23) mm by disk diffusion and 18.3 ± 1.3 (15 - 20) mm, 19.9 ± 2.6 (18 - 22) mm, respectively, by agar well diffusion methods for C. albicans. On the other hand, ABS and AMB showed inhibition zones with mean diameters of 19.4 ± 1.5 (18 - 24) mm, 19.1 ± 2.8 (13 - 30) mm by disk diffusion and 19.8 ± 2.1 (18 - 25) mm, 18.7 ± 2.3 (13 - 23) mm by agar well diffusion methods for non-albicans Candida isolates. ABS exhibited higher activity against non-albicans Candida species compared to C. albicans (P < 0.001). By the time kill-curve method, ABS achieved a 4 log10 cfu/mL decrease in C. albicans ATCC 90028, C. parapsilosis ATCC 22019, and C. krusei ATCC 6258, as well as seven different Candida spp. isolates of C. albicans, C. glabrata, C. tropicalis, C. guilliermondii, C. kefyr, C. krusei, and C. parapsilosis, respectively. Conclusions: ABS can be an alternative for treating superficial infections.

Missense mutation in CgPDR1 regulator associated with azole-resistant Candida glabrata recovered from Thai oral candidiasis patients.

Non-albicans Candida (NAC) species are increasingly identified as pathogens causing oral candidiasis. Incidence rates for azole resistance among NAC species have been continuously reported. This study aimed to evaluate the azole susceptibility profiles and to characterise the azole resistance mechanisms of oral clinical NAC isolates. In vitro susceptibility patterns of 85 NAC species isolates were determined by the broth microdilution method. Azole resistance-related genes (ERG3, ERG11 and PDR1) of Candida glabrata isolates were sequenced to determine the presence of nucleotide substitutions. Expression levels of various resistance-related genes were also evaluated by RT-qPCR in azole-susceptible, susceptible dose-dependent (SDD) and resistant Candida isolates. Two C. glabrata isolates (2.4% of all NAC isolates) were resistant to all three azoles tested (fluconazole, itraconazole and ketoconazole). All clinical isolates of Candida tropicalis and Candida kefyr were susceptible to azoles. Silent mutations were found in the CgERG11 and CgERG3 genes of clinical C. glabrata isolates. Interestingly, two missense mutations in CgPDR1 (N768D and E818K) were identified only in resistant C. glabrata isolates. The presence of a CgPDR1 missense mutation in resistant isolates is associated with overexpression of its own product as well as multidrug transporters including ABC and MFS transporters. A gain-of-function (GOF) mutation in CgPDR1 is associated with upregulation of various drug transporters, which appears to serve as a primary mechanism for azole resistance in the detected C. glabrata isolates. Therefore, analysis of GOF mutations in the PDR1 regulator provides a better understanding of the development of antifungal resistance.

Distribution and antifungal susceptibility profile of Candida species from candiduria cases at a tertiary care hospital

The present study was conducted to assess the distribution of Candida species in candiduria and to evaluate their antifungal susceptibility pattern. Urine samples were subjected to microscopy and screened for the growth. Candida isolates were identified and speciated by colour on HiCHROM Candida agar, germ tube test, growth in cornmeal agar to study micromorphology, carbohydrate assimilation and fermentation tests according to standard protocols. All isolates were investigated for antifungal susceptibility testing. During the study period, a total number of 2560 urine samples were reported. Out of them 94/2560 (3.7%) urine samples showed significant candiduria. 60 from catheterised patients admitted to the ICU and 34 from patients admitted in different wards. The species-wise distribution among Candida isolates are C.tropicalis53 (56.4%), C.albicans21(22.4%), C.glabrata8 (8.5%), C. krusei 7 (7.4%) and C.parapsilosis 5 (5.3%). Candida species exhibited resistance to Amphotericin B 6 (6.4%), Fluconazole 23 (24.5%) Voriconazole 8 (8.5%), Flucytosine 5 (5.3%) and 4 (4.3%) to Caspofungin.Candidapreviously presumed as non-pathogenic has attained an important clinical role in nowadays circumstances of growing risk factors and developing antifungal resistance. The frequency of isolation of non-albicans Candida was more when compared to Candida albicans and they showed more resistance to antifungal drugs when compared to C.albicans.Caution must be taken in reporting Candida from urine, but it is recommended not to mistreat candiduria. The Candida species identification along with their antifungal susceptibility profile can assist clinicians in choosing better antifungal drugs for the treatment of patients with candiduria.

Activity of Diosgenyl 2-amino-2-deoxy-β-D-glucopyranoside, its Hydrochloride, and N,N-dialkyl Derivatives Against Non-albicans Candida Isolates.

Candida albicans belongs to the most common fungal pathogens in humans, but recently an increased proliferation of strains called non-albicans Candida has been reported. Species belonging to this group are often characterised by a reduced susceptibility to antifungal agents. In view of the emergence of non-albicans Candida and their resistance to available antifungals, an attempt has been made to develop novel effective agents. Biological activities of the N,N-dialkyl diosgenyl glycosides, which were previously synthesized, were determined. Minimum inhibitory concentration (MIC) was determined for group of clinical nonalbicans Candida isolates by serial dilution method in Sabouraud liquid medium. In order to assess the toxicity towards human cells the minimum haemolytic concentration (MHC) was determined on human erythrocytes by serial dilution method in phosphoric buffer. The saponins exhibited a strong activity towards clinical isolates of C. glabrata and C. parapsilosis comparable or even stronger than that of conventional antimicrobials. A high rate of resistance to fluconazole was shown among C. glabrata isolates. Among clinical strains of C. krusei and C. tropicalis, isolates with a decreased susceptibility to saponins were identified. All the tested C. krusei isolates showed resistance to fluconazole, while among C. tropicalis numerous strains were resistant to all tested azoles. The saponins did not show haemolytic activities at their microbiologically active concentrations. Results of the present work encourage to continue the study on steroidal saponins and their potential application for the treatment of candidemia.

In Vitro Susceptibilities of Non-albicans Candida Species to Echinocandins, Azoles, and Amphotericin B in Tokat, Turkey

Background: Prophylactic and therapeutic uses of antifungal agents have given rise to a significant shift to more resistant non-albicans Candida species associated with fungal infections. Objectives: This study aimed at identifying the distribution and antifungal susceptibility patterns of non-albicans Candida spp. isolated from clinical specimens in Tokat, Turkey. Methods: The authors determined the susceptibility of 103 non-albicans Candida isolates to the following antifungal agents: amphotericin B, anidulafungin, caspofungin, fluconazole, ketoconazole, itraconazole, voriconazole, and posaconazole, using the Etest method. Interpretation of susceptibility was carried out using species specific breakpoints suggested by the Clinical and Laboratory Standards Institute (CLSI) M27-S4 document. Results: The most frequently isolated non-albicans Candida species were Candida kefyr (44 isolates, 42.8%) followed by C. tropicalis (36 isolates, 35%), C. parapsilosis (17 isolates, 16.5%), C. glabrata (four isolates, 3.8%) and C. famata (two isolates, 1.9%). None of the strains had MIC values of > 2 µg/mL for amphotericin B except three of the 44 C. kefyr isolates. Resistance to caspofungin and anidulafungin were not detected in C. tropicalis, C. parapsilosis, and C. glabrata isolates. Only two of the 36 C. tropicalis isolates were categorized as intermediate resistant to anidulafungin, according to the new CLSI criteria. None of the C. parapsilosis isolates were found to be resistant to azole drugs. Conclusions: Most of the non-albicans Candida species were found to be susceptible to tested antifungal drugs. Therefore, use of routine antifungal agents like amphotericin B and fluconazole, which are available in this region, are suggested.

Extracts From Hypericum hircinum subsp. majus Exert Antifungal Activity Against a Panel of Sensitive and Drug-Resistant Clinical Strains.

During the last two decades incidences of fungal infections dramatically increased and the often accompanying failure of available antifungal therapies represents a substantial clinical problem. The urgent need for novel antimycotics called particular attention to the study of natural products. The genus Hypericum includes many species that are used in the traditional medicine to treat pathological states like inflammations and infections caused by fungi. However, despite the diffused use of Hypericum-based products the antifungal potential of the genus is still poorly investigated. In this study five Hypericum species autochthonous of Central and Eastern Europe were evaluated regarding their polyphenolic content, their toxicological safety and their antifungal potential against a broad panel of clinical fungal isolates. LC-MS analysis led to the identification and quantification of 52 compounds, revealing that Hypericum extracts are rich sources of flavonols, benzoates and cinnamates, and of flavan-3-ols. An in-depth screen of the biological activity of crude extracts clearly unveiled H. hircinum subsp. majus as a promising candidate species for the search of novel antifungals. H. hircinum is diffused in the Mediterranean basin from Spain to Turkey where it is traditionally used to prepare a herbal tea indicated for the treatment of respiratory tract disorders, several of which are caused by fungi. Noteworthy, the infusion of H. hircinum subsp. majus excreted broad antifungal activity against Penicillium, Aspergillus and non-albicans Candida isolates comprising strains both sensitive and resistant to fluconazole. Additionally, it showed no cytotoxicity on human cells and the chemical characterization of the H. hircinum subsp. majus infusion revealed high amounts of the metabolite hyperoside. These results scientifically support the traditional use of H. hircinum extracts for the treatment of respiratory tract infections and suggest the presence of exploitable antifungal principles for further investigations aimed at developing novel antifungal therapies.

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.

Fluconazole Resistance among Oral Candida Isolates from People Living with HIV/AIDS in a Nigerian Tertiary Hospital.

Oropharyngeal candidiasis, a common fungal infection in people living with HIV/AIDS (PLWHA), arises from Candida species colonizing the oral cavity. Fluconazole is the preferred treatment and is often used empirically. Few studies have investigated the prevalence of fluconazole resistance in Nigeria. This study aimed at determining the burden of fluconazole resistance among Candida species in the oral cavities of PLWHA. We sampled the oral cavities of 350 HIV-infected adults and an equal number of HIV-negative controls. Candida isolates were identified using germ tube tests, CHROMagar Candida (CHROMagar, Paris, France), and API Candida yeast identification system (BioMérieux, Marcy-l’Étoile, France). Fluconazole susceptibility was determined using the Clinical and Laboratory Standards Institute disc diffusion method. Data were analysed using SPSS version 21 (IBM, New York, NY, USA). The significance level was set at p ≤ 0.05. The isolation rates for Candida amongst HIV-infected subjects and controls were 20.6% and 3.4%, respectively (p < 0.001). In PLWHA, Candida albicans was most frequently isolated (81.3%) and fluconazole resistance was present in 18 (24%) of the 75 Candida isolates. Resistance to fluconazole was present in half of the non-albicans Candida isolates. Fluconazole resistance is prevalent among oral Candida isolates in PLWHA in the study area with a significantly higher rate among non-albicans Candida spp.

Profile of fungal septicaemia in new born at a tertiary care hospital in North India

Background: Advances in neonatal management have led to considerable improvement in new-born survival. The objective of this study was to determine the predominant organisms, antifungal sensitivity patterns and clinical risk factors in neonatal fungal blood stream infection cases (BSI) admitted to our hospital.Methods: This is a retrospective study of all neonatal fungal BSI cases between January 2015 to December 2015.Results: Fungal sepsis was found in 50/360 (13.6%) of cases. Non Albicans candida (NAC)species were responsible for 88% of cases with Candida. glabrata (54%)as the most predominant species. Other species isolated were C. tropicalis 9 (18%). C. albicans 6 (12%). C. paraspinolosis 5 (10%), C. Krusei 2 (4%) and C. Kodo1 (2%). Antifungal sensitivity results revealed that most of the NAC isolates especially candida glabrata,candida paraspinolosis were resistant to flucanazole, than Candida albicans. Amphoterician B had greater sensitivity than FLK over NAC species Among the risk factor observed for candidemia were low birth weight (62%), prematurity (60%,), broad spectrum antibiotic use (60%), ventilater support (56%) and total parentral nutrition (50%).Conclusions: The increase in neonatal fungal BSI and resistant organisms highlights the need to review use of strict infection control strategies, appropriate preventive and therapeutic measures such as prophylactic antifungal use and a restrictive policy of antibiotic use.

In vitro synergism of a water insoluble fraction of Uncaria tomentosa combined with fluconazole and terbinafine against resistant non-Candida albicans isolates

Context: Uncaria tomentosa D.C. (Rubiaceae) has several biological activities, including activity against resistant Candida strains. The synergistic interaction with terbinafine or fluconazole can be an important alternative to overcome this resistance.Objectives: The potential synergy between a water insoluble fraction (WIF) from Uncaria tomentosa bark and the antifungals terbinafine (TRB) and fluconazole (FLZ) against non-Candida albicans resistant strains was investigated.Materials and methods: TRB and FLZ, alone and combined with WIF, were tested by the checkerboard procedure using the micro-dilution technique against seven isolates of Candida glabrata and C. krusei. The molecular interactions occurring outside the cell wall were evaluated by scanning electron microscopy, Fourier transform infrared (FT-IR) and differential scanning calorimetry (DSC) analysis.Results: The checkerboard inhibitory assay demonstrated synergy for WIF:TRB and WIF:FLZ combinations, respectively. The best synergistic cell damage was demonstrated unequivocally for the associations of WIF and TRB (1.95:4.0 μg/mL) and WIF and FLZ (1.95:8.0 μg/mL). The comparison of the FT-IR spectra of the antifungal alone, and in combination with WIF, allows recognizing clear differences in 3000, 1600, 1400, and 700–800 cm−1 bands. Additionally, modifications on TRB and FLZ thermograms were clearly noticed after their combination with WIF.Conclusions: DSC and infrared analysis demonstrated intermolecular interactions between WIF and either TRB or FLZ. Hence, quite likely the synergistic effect is related to interaction events occurring outside the cell wall between antifungal and cat’s claw proanthocyanidins. A direct action on the cell wall is suggested, without connection with the ABC efflux pump mechanism.

Candida species from oral cavity of HIV-infected children exhibit reduced virulence factors in the HAART era.

This study aimed to assess, invitro, the biofilm viability and the phospholipase and protease production of Candida spp. from the saliva of HIV infected children and healthy controls, and to correlate the results with the use of medical data. A total of 79 isolates were analyzed: 48 Candida albicans isolates (33/15) and 20 Candida parapsilosis sensu lato complex isolates (12/8) (from HIV/control patients, respectively), and 8 Candida krusei, 1 Candida tropicalis, 1 Candida dubliniensis and 1 Candida guilliermondii from HIV patients. The XTT (2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-Carboxanilide) reduction assay analyzed the biofilm viability. Phospholipase and protease assays were performed using the egg yolk and Bovine Serum Albumin agar plate methods, respectively. All isolates were able to form biofilm with cell viability. Quantitatively, Candida isolates from both groups presented a similar ability to form biofilm (p>0.05). The biofilm viability activity was higher in C.albicans isolates than in non-albicans Candida isolates (p<0.05) for both groups. Phospholipase activity was detected in 32 isolates (40.5%) and it was significantly higher in the HIV group (p=0.006). Protease activity was detected in 66 isolates (84.8%) and most of them were relatively/very strong producers. No statistical association with medical data was found in the HIV group. Although Candida spp. isolates from HIV-positive children presented higher phospholipase production, invitro they exhibited reduced virulence factors compared to isolates from healthy individuals. This finding may enlighten the role played by immunosuppression in the modulation of Candida virulence attributes.

Phenotypic variability and therapeutic implications of Candida species in patients with oral lichen planus

We investigated the prevalence and phenotypic variation of Candida species in oral lichen planus (OLP) and the therapeutic implications of our findings. Eighty patients with clinically and histopathologically confirmed cases of OLP (64 non-erosive, 16 erosive) and a control group of 80 healthy individuals with no predisposing factors for oral candidiasis were examined for evidence of Candida infection. Oral swabs and smears were obtained for cytology and culture. Identification, speciation and antifungal susceptibility tests of Candida isolates were performed using an automated microbial identification system. Fifty percent of erosive OLP cases, 28% of non-erosive cases and none of the controls showed evidence of Candida. Candida albicans was found predominantly in non-erosive OLP, while other Candida species were predominate in erosive OLP. Non-Candida albicans isolates (C. glabrata, C. krusei) were resistant to the commonly used antifungals, clotrimazole and fluconazole. Candida infection is common in cases of OLP. We recommend antifungal sensitivity testing prior to antifungal therapy for the erosive form of OLP.

Comparison Between Biofilm Production, Phospholipase and Haemolytic Activity of Different Species of Candida Isolated from Dental Caries Lesions in Children.

C.albicans is the most commonly isolated fungal pathogen in the oral cavity, but isolation of non-albicans Candida is increasing in recent years. We wish to demonstrate the virulence factors of Candida spp. isolated from the dental caries lesion of the children as presence of virulence factors determines the pathogenic potential of any microorganism. To compare biofilm production, phospholipase and haemolytic activity of C.albicans with that of non-albicans species of Candida isolated from dental caries lesions of children to evaluate the role of non- albicans species of Candida in formation of dental caries. Oral swabs were collected from caries lesion of 100 school children of age 5-10 years with dental caries. Candida isolates were tested for biofilm production, phospholipase and haemolytic activity. Statistical analysis was done by Chi-Square test and Mann-Whitney U test wherever applicable using SPSS version 11.5. Out of the 100 children with dental caries 37 were positive for Candida by smear or culture and 31 by culture. C.albicans was the most prevalent isolate followed by C.krusei, C.tropicalis and C.albicans. Out of 21 C.albicans isolates, 10 (47.6%) showed phospholipase activity and 18 (85.71%) produced biofilm. Of the 10 non-albicans strains, 5 (50%) showed phospholipase activity and 6 (60%) produced biofilm. All isolates of Candida produced haemolysin (100%). There was no statistically relevant difference between the virulence factor production by C.albicans and non-albicans species of Candida. In other words, our study shows that both C.albicans and non-albicans species of Candida isolated from caries lesions of the children, produce these virulence factors. So we can say that non-albicans species of Candida also are involved in caries formation.

Antifungal susceptibility pattern of Candida species isolated from suspected cases of tuberculosis

Background: Candida species are emerging as a potentially pathogenic fungus in patients with broncho-pulmonary diseases. The synergistic growth promoting association of Candida and Mycobacterium tuberculosis has raised increased concern for studying the various Candida spp. and its significance in tuberculosis patients during current years. Aim: Aim of our study was isolation and identification of Candida spp from tuberculosis patients; as well as testing sensitivity of these clinical isolates to commonly prescribed antifungal drugs. Method: Totally 178 sputum specimens were screened by RNTCP Unit were included in the study. In vitro susceptibility testing of C. albicans isolated from pulmonary tuberculosis patients to five antifungal drugs was carried by direct susceptibility method using CHROM agar. Results: We could able to obtain 20.2% of Candida strains from 178 RNTCP clinic attendees and Candida albicans, Candida parapsilosis, Candida dubliniensis and Candida krusei was identified. Overall based on our study report we could able to report 33.3%,19.4% and 8.3% antifungal resistance by Candida strains towards the antifungal drugs Itraconazole, Nystatin, and Amphotericin B, respectively. Conclusion: In this study we found a shifting pattern of epidemiology of Candida species from commensal to emerging pathogen. Therefore, screening of tuberculosis patients for Candida infection should be routinely practiced along with anti-fungal sensitivity testing for non-albicans Candida isolates. Keywords: Antifungal susceptibility, azoles, Candida, disk diffusion, tuberculosis, Non-albicans Candida(NAC)