Potential Inhibitory Effect of Miltefosine against Terbinafine-Resistant Trichophyton indotineae.

Several prolonged and significant outbreaks of dermatophytosis caused by Trichophyton indotineae, a new emerging terbinafine-resistant species, have been ongoing in India in recent years, and have since spread to various countries outside Asia. Miltefosine, an alkylphosphocholine, is the most recently approved drug for the treatment of both visceral and cutaneous leishmaniasis. Miltefosine in vitro activity against terbinafine-resistant and susceptible T. mentagrophytes/T. interdigitale species complex, including T. indotineae, is limited. The current study aimed to assess miltefosine's in vitro activity against dermatophyte isolates, which are the most common causes of dermatophytosis. Miltefosine, terbinafine, butenafine, tolnaftate, and itraconazole susceptibility testing was performed using Clinical and Laboratory Standards Institute broth microdilution methods (CLSI M38-A3) against 40 terbinafine-resistant T. indotineae isolates and 40 terbinafine-susceptible T. mentagrophytes/T. interdigitale species complex isolates. Miltefosine had MIC ranges of 0.063-0.5 µg/mL and 0.125-0.25 µg/mL against both terbinafine-resistant and susceptible isolates. In terbinafine-resistant isolates, the MIC50 and MIC90 were 0.125 µg/mL and 0.25 µg/mL, respectively, and 0.25 µg/mL in susceptible isolates. Miltefosine had statistically significant differences in MIC results when compared to other antifungal agents (p-value 0.05) in terbinafine-resistant strains. Accordingly, the findings suggest that miltefosine has a potential activity for treating infections caused by terbinafine-resistant T. indotineae. However, further studies are needed to determine how well this in vitro activity translates into in vivo efficacy.

Antifungal activity of miltefosine against both azole-susceptible and -resistant Aspergillus strains.

Miltefosine, an alkylphosphocholine, has been approved recently for the treatment of visceral leishmaniasis. Miltefosine has shown promise as a treatment for paracoccidioidomycosis, and has mixed activity against other fungi and yeast. There are limited data on the in-vitro activity of miltefosine against azole-resistant and -susceptible Aspergillus spp. As such, the aim of this study was to determine the in-vitro activity of miltefosine against Aspergillus strains. Miltefosine was tested against 108 azole-susceptible and -resistant Aspergillus strains isolated from Iran and other countries using the broth microdilution method. Miltefosine was found to be effective against azole-resistant Aspergillus isolates, with minimum inhibitory concentrations (MICs) ranging from 1.562 to 6.25 µg/mL. MIC50 and MIC90 were 1.562 and 3.125 µg/mL, respectively. Miltefosine had a higher geometric mean MIC (2.459 µg/mL) for wild-type Aspergillus isolates than itraconazole (0.220 µg/mL) and voriconazole (0.298 µg/mL). No significant difference was found between miltefosine MICs for azole-resistant Aspergillus isolates and azole-susceptible Aspergillus isolates (P>0.05). Miltefosine appears to have good in-vitro activity against azole-resistant Aspergillus strains, according to these findings. Furthermore, the findings suggest that miltefosine could be used to treat infections caused by azole-resistant Aspergillus spp.

Iranian National Survey on Tinea Capitis: Antifungal Susceptibility Profile, Epidemiological Characteristics, and Report of Two Strains with a Novel Mutation in SQLE Gene with Homology Modeling.

BACKGROUND: The data on the epidemiological and antifungal susceptibility profile of tinea capitis (TC) in Iran has not been updated in recent decades. This report presents the Iranian epidemiological and drug susceptibility data regarding the distribution of dermatophytes species isolated by six national mycology centers for a period of one year (2020-2021). MATERIAL AND METHODS: A total of 2100 clinical samples from individuals suspeted to TC were subjected to mycological analysis of direct microscopy and culture. For definite species identification, the culture isolates were additionally subjected to PCR-RFLP and PCR-sequencing of the ITS ribosomal DNA (ITS-rDNA) region. Antifungal susceptibility profiles for eight common antifungal drugs were determined by CLSI M38-A3 guidelines. The SQLE gene was partially amplified and sequenced in two terbinafine-resistant and two susceptible T. mentagrophytes isolates to elucidate probable substitutions involved in resistance. RESULTS: TC (n = 94) was diagnosed in 75 children (79.8%) and 19 adults (20.2%) by direct microscopy and culture. Frequency of TC was significantly more among males (66 males = 70.2% vs 28 females = 29.8%). The prevalent age group affected was 5-9 years (39.36%). Thirty-two (34.04%) T. mentagrophytes, 27 (28.7%) T. tonsurans, 14 (14.9%) M. canis, 13 (13.8%) T. violaceum, 5 (5.32%) T. indotineae, 2 (2.1%) T. benhamiae, and 1 (1.1%) T. schoenleinii were identified as the causative agents. MIC values of isolates showed susceptibility to all antifungal agents, except for fluconazole and griseofulvin with GM MIC of 11.91 µg/ml and 2.01 µg/ml, respectively. Terbinafine exhibited more activity against isolates, with GM MIC 0.084 µg/ml followed by ketoconazole (0.100 µg/ml), econazole (0.107 µg/ml), itraconazole (0.133 µg/ml), butenafine (0.142 µg/ml), and miconazole (0.325 µg/ml). Two resistant T. mentagrophytes isolates harbored missense mutations in SQLE gene, corresponding to amino acid substitution F397L. Remarkably, one unique mutation, C1255T, in the SQLE sequence of two terbinafine-susceptible T. mentagrophytes strains leading to a change of leucine at the 419th position to phenylalanine (L419F) was detected. CONCLUSIONS: T. mentagrophytes, T. tonsurans, and M. canis remained the main agents of TC in Iran, however less known species such as T. indotinea and T. benhamiae are emerging as new ones. Terbinafine could still be the appropriate choice for the treatment of diverse forms of TC.

In Vitro Combination of Terbinafine with Ketoconazole Against Aspergillus Species with Terbinafine High MIC Values Isolated From Otomycosis.

Otomycosis is a common mycotic infection of the external auditory canal, and Aspergillus species are one of the most frequent causative agents worldwide. The limited antifungal arsenal, the high toxicity and side effects of antifungal agents, and the growing resistance to the currently available antifungals underscore the need for new therapeutic strategies. The present study aimed to evaluate the combined in vitro efficacy of terbinafine and ketoconazole against Aspergillus species with terbinafine high MIC values isolated from patients with otomycosis.84 Aspergillus species with high MIC values to terbinafine (≥ 4 µg/ml), consisting of A. flavus, A. tubingensis, A. niger, and A. terreus, were included in this study. The checkerboard microdilution method evaluated the in vitro interactions using the CLSI reference technique. Synergistic effects were observed for 66.67% (56/84) of all isolates (FICI ranging from 0.19 to 0.5). However, the interactions of terbinafine and ketoconazole exhibited indifference in 33.33% (28/84) of the isolates, and no antagonism was observed for any combination. The interaction of terbinafine and ketoconazole showed synergistic activity against Aspergillus species with high MIC values, suggesting that this is an alternative and promising approach for treating otomycosis.

In vitro antifungal susceptibility profile of Iranian Fusarium isolates: Emphasising on the potent inhibitory effect of efinaconazole compared to other drugs.

BACKGROUND: Fusarium species are opportunistic human pathogens that remarkably cause fungal infections ranging from superficial to fatal invasive disseminated infections. Fusarium species are notoriously resistant to the majority of antifungal agents. OBJECTIVES: Therefore, detailed studies regarding in vitro susceptibility are required and may lead to a better prognosis of severe infections. METHODS: We evaluated 25 antifungal drugs in vitro against 282 clinical and environmental Fusarium isolates. RESULTS: Fusarium species demonstrated high MICs/MECs values to the most commonly used antifungal drugs in clinical practice. The geometric mean (GM) MICs for luliconazole (0.004 µg/ml) and lanoconazole (0.012 µg/ml) were the lowest, followed by efinaconazole (0.98 µg/ml) and amphotericin B (1.04 µg/ml). CONCLUSIONS: Efinaconazole, a novel triazole, may be a promising candidate for the treatment of superficial Fusarium infections. Furthermore, the development of systemic formulations of these drugs as well as further in vitro and in vivo investigations could aid in the treatment of systemic fusariosis.

In Vitro Antifungal Susceptibility Profile of Miltefosine against a Collection of Azole and Echinocandins Resistant Fusarium Strains.

Fusarium species are filamentous fungi that cause a variety of infections in humans. Because they are commonly resistant to many antifungal drugs currently available in clinical settings, research into alternative targets in fungal cells and therapeutic approaches is required. The antifungal activity of miltefosine and four comparators, amphotericin B, voriconazole, itraconazole, and caspofungin, were tested in vitro against a collection of susceptible and resistant clinical (n = 68) and environmental (n = 42) Fusarium isolates. Amphotericin B (0.8 µg/mL) had the lowest geometric mean (GM) MICs/MECs values followed by miltefosine (1.44 µg/mL), voriconazole (2.15 µg/mL), caspofungin (7.23 µg/mL), and itraconazole (14.19 µg/mL). Miltefosine was the most effective agent against Fusarium isolates after amphotericin B indicating that miltefosine has the potential to be studied as a novel treatment for Fusarium infections.

In vitro activity of 23 antifungal drugs against 54 clinical and environmental Aspergillus oryzae isolates.

The treatment of invasive aspergillosis caused by cryptic species remains a challenge due to the lack of randomised clinical trials and investigation of the efficacy and safety of different therapeutic strategies. We aimed to evaluate the in vitro activity of 23 conventional and new antifungal drugs against 54 clinical and environmental Aspergillus oryzae isolates by using the Clinical and Laboratory Standards Institute (CLSI) standard M38-A3. The lowest geometric mean MIC values were found for luliconazole and lanoconazole (0.001 µg/ml), followed by anidulafungin (0.104 µg/ml), posaconazole (0.15 µg/ml), itraconazole (0.37 µg/ml), efinaconazole (0.5 µg/ml), voriconazole (0.51 µg/ml), tavaborole (0.72 µg/ml), and amphotericin B (0.79 µg/ml). In contrast, ketoconazole, terbinafine, econazole, tioconazole, ravuconazole, miconazole, nystatin, clotrimazole, griseofulvin, sertaconazole, natamycin, tolnaftate, and fluconazole had no or low activity. Further studies are required to determine how well this in vitro activity translates into in vivo efficacy.

First report of tinea corporis caused by Trichophyton quinckeanum in Iran and its antifungal susceptibility profile.

Background and Purpose: Trichophyton quinckeanum, a known zoophilic dermatophyte responsible for favus form in rodents and camels, is occasionally reported to cause human infections. Case Report: This study aimed to report a case of tinea corporis caused by T. quinckeanum that experienced annular erythematous pruritic plaque with abundant purulent secretions. In June 2021, a 15-year-old girl with an erythematous cup shape lesion on the right wrist bigger than 3 cm in diameter was examined for tinea corporis. Since March, 2016 her family has kept several camels at home. Direct examination of skin scraping and purulent exudates revealed branching septal hyaline hyphae and arthrospore. Morphological evaluation of the recovered isolate from the culture and sequencing of ITS1-5.8S rDNA-ITS2 region resulted in the identification of T. quinckeanum. Antifungal susceptibility testing showed that this isolate had low minimum inhibitory concentration (MIC) values for luliconazole, terbinafine, and tolnaftate, but high MICs to itraconazole, fluconazole, posaconazole, miconazole, isavuconazole, ketoconazole, clotrimazole, and griseofulvin. However, the patient was successfully treated with oral terbinafine and topical ketoconazole. Conclusion: It can be said that T. quinckeanum is often missed or misidentified due to its morphological similarity to T. mentagrophytes/T. interdigitale or other similar species. This dermatophyte species is first reported as the cause of tinea corporis in Iran. As expected, a few months after our study, T. quinckeanum was detected in other areas of Iran, in a few cases.

Phylogenetic Analysis of Clinically Relevant Fusarium Species in Iran.

Fungi of the genus Fusarium are well known as major plant pathogens but also cause a broad spectrum of human infections. Sixty-three clinical isolates, collected during 2014-2017, were identified using a part of the TEF1 gene as barcoding marker. Fusarium fujikuroi species complex (FFSC, n = 41, 65%) showed to be the dominant etiological agent, followed by F. solani species complex (FSSC, n = 14, 22%) and F. oxysporum species complex (FOSC, n = 7, 11%). There was one strain belonging to F. lateritium species complex (FLSC, n = 1, 1.5%). For final identification, a phylogenetic tree was constructed including the type strains of each species complex. Most cases of fusariosis were due to nail infection (n = 38, 60.3%), followed by keratitis (n = 22, 34%). Fusarium infections are difficult to be treated due to their intrinsic resistance to different azoles; however, accurate and fast identification of etiological agents may enhance management of the infection. We present the first phylogenetic study on clinical Fusarium spp. from Iran.

Molecular epidemiology of Tinea gladiatorum in contact sports in northern Iran.

BACKGROUND: Tinea gladiatorum as the major health concern among combat athletes and the most common cause of a decrease in the quality of their sports activities. OBJECTIVES: This study aimed to determine the molecular epidemiology of tinea gladiatorum in contact sports in Mazandaran Province, Northern Iran. MATERIALS/METHODS: In total, 631 suspected athletes out of 4240 ones were screened for tinea gladiatorum in this cross-sectional study from May 2018 to February 2019. The infection was confirmed by direct microscopy and culture tests. The causative agents were identified by polymerase chain reaction-restriction fragment length polymorphism method on ITS rDNA region. The indoor environmental factors were evaluated in 81 sports halls in terms of temperature (°C), humidity (%), density (m2 /athletes) and the number of ventilators. RESULTS: Tinea gladiatorum was confirmed in 6.5% (278/4240) of the athletes. Tinea corporis was the most common type of tinea (192/278; 69.0%), and 175 cases (27.7%) had more than one type of tinea. Moreover, Trichophyton tonsurans (192/203; 94.5%) was the most aetiological agent in all athletes followed by Microsporum canis (6/203; 2.9%), Trichophyton rubrum (2/203; 0.9%) and Trichophyton interdigitale (1/203; 0.4%). There was no significant association between the prevalence of tinea gladiatorum and the indoor environmental factors. CONCLUSION: T. tonsurans is the most common agent of tinea gladiatorum in Northern Iran. Since no positive culture of dermatophyte was obtained from mats, it seems that skin-to-skin contact would be the most probable mode of transmission among wrestlers. Accordingly, periodical checkups should be considered by health care providers.

High prevalence of asymptomatic nosocomial candiduria due to Candida glabrata among hospitalized patients with heart failure: a matter of some concern?

BACKGROUND AND PURPOSE: Heart failure is a leading cause of hospitalization, and asymptomatic candiduria is common in hospitalized patients with low morbidity. However, in most patients, it is resolved spontaneously on the removal of the catheter. Despite the publication of guidelines, there are still controversies over the diagnosis and management of candiduria. However, in hospitalized patients with heart failure, the decision to treat candiduria is especially important since the nosocomial infections are associated with an increase in morbidity, mortality, length of hospital stay, and healthcare costs. Some species of Candida, such as Candida glabrata, are increasingly resistant to the first-line and second-line antifungal medications. The present study aimed to investigate the incidence of asymptomatic Candida urinary tract infection due to C. glabrata and antifungal susceptibility of Candida isolates in hospitalized patients with heart failure. MATERIALS AND METHODS: In total, 305 hospitalized patients with heart failure were studied to identify asymptomatic nosocomial candiduria during 2016-17 in one private hospital in the north of Iran. The Sabouraud's dextrose agar culture plates with a colony count of >104 colony-forming unit/ml of urine sample were considered as Candida urinary tract infection. Candida species were identified based on the morphology of CHROMagar Candida (manufactured by CHROMagar, France) and PCR-RFLP method with MspI restriction enzyme. Antifungal susceptibility testing of the isolates was performed using five mediations, including itraconazole, voriconazole, fluconazole, amphotericin B, and caspofungin by broth microdilution method according to CLSI M27-S4. RESULTS: In this study, the rate of asymptomatic Candida urinary tract infection was 18.8%, which was more common in people above 51 years old and females (70%). In addition to the urinary and intravascular catheter, the occurrence of candiduria in hospitalized patients had significant relationships with a history of surgical intervention, diastolic heart failure, and use of systemic antibiotics (P>0.05). Among Candida spp., non-albicans Candida species was the most common infectious agent (59.7%). Moreover, C. glabrata (n=27, 40.3%) (alone or with other species) and Candida albicans (n=27, 40.3%) were the most common agents isolated in Candida urinary tract infection. Based on the results of the in vitro susceptibility test, the C. glabrata isolates were 15%, 59%, 70%, 74%, and 85% susceptible to caspofungin, amphotericin B, itraconazole, voriconazole, and fluconazole, respectively. CONCLUSION: According to the findings, there was a high prevalence of asymptomatic Candida urinary tract infection in hospitalized patients with heart failure. Besides, it was suggested that there was a shift towards non-albicans Candida, especially C. glabrata, in these patients. Therefore, asymptomatic candiduria in hospitalized patients with heart failure should be considered significant. Furthermore, the identification of Candida species along with antifungal susceptibility is essential and helps the clinicians to select the appropriate antifungal agent for better management of such cases.

Fatal rhinocerebral mucormycosis in a patient with ulcerative colitis receiving azathioprine and corticosteroid.

BACKGROUND AND PURPOSE: Rhinocerebral mucormycosis is a rare fatal fungal infection which is on a growing trend, particularly among immunocompromised patients. Immunosuppressive drugs, including corticosteroids and antimetabolites, increase the risk of this infection. Herein, we reported the case of fulminant rhinocerebral mucormycosis in a patient with ulcerative colitis receiving azathioprine and corticosteroid. CASE REPORT: A 58-year-old woman was admitted to the hospital in a state of coma with an extensive necrosis in her nose. She was afflicted with intestinal bleeding after 1 month of fasting and was treated with azathioprine and a high dose of prednisolone for ulcerative colitis 2 months prior to hospital admission. The direct microscopic examination of the necrotic tissues of the paranasal sinuses showed several non-septate hyphae consistent with Mucorales. Culture media yielded Rhizopus species, which was identified as Rhizopus oryzae by internal transcribed spacer polymerase chain reaction sequencing. Despite the implementation of surgical and pharmaceutical (liposomal amphotericin B) treatments, the patient expired after 2 weeks of admission. CONCLUSION: The gastroenterologists should be aware of the adverse effect of immunosuppressive drugs they prescribe for the treatment of inflammatory bowel disease.

Catheter-related candidemia and identification of causative Candida species in patients with cardiovascular disorder.

BACKGROUND AND PURPOSE: Catheter-related blood circulation infection is the most dangerous and serious side-effects of vascular catheters, which leads to the enhancement of the costs, mortality, and hospital stay duration, especially in the Intensive Care Unit. Regarding this, the aim of the current study was to identify the prevalence of catheter-induced candidemia in the Tehran Heart Center, a heart hospital in Tehran, Iran. MATERIALS AND METHODS: This study was conducted on patients admitted to Tehran Heart Center for a minimum of 7 days during 18 months. To detect the fungal elements, blood culture and catheter culture were performed in the patients receiving central or peripheral venous catheter. Then, the polymerase chain reaction (PCR) was applied to determine the possible diagnosis. RESULTS: The investigation of 223 samples led to the identification of a total of 15 (6.7%) yeast isolates obtained from 9 (60%), 4 (26.6 %), and 2 (13.4%) catheter, blood, and skin (of the catheter insertion areas) cultures, respectively. Out of nine Candida isolates obtained from the catheter samples, 1 (11.1%), 1 (11.1%), 2 (22.2%), and 5 (55.6%) cases were identified as C. tropicalis, C. membranifaciens, C. glabrata, and C. albicans, respectively, using the internal transcribed spacer region sequencing. Furthermore, the four yeasts isolated from the blood culture included C. tropicalis, C. carpophila, C. membranifaciens, and Cryptococcus albidus. Additionally, one case of C. glabrata and one case of C. albicans were isolated from the skin culture of the catheter insertion areas in patients with positive catheter culture. We reported two cases of catheter-related candidemia caused by C. membranifaciens and C. tropicalis on the basis of the genetic similarity of the species isolated from the blood and catheter. These cases were treated successfully with intravenous fluconazole and catheter removal. CONCLUSION: There is some evidence indicating the growing prevalence of non-albicans Candida infections. Many risk factors, including prior antibiotic therapy, use of a central venous catheter, surgery, and parenteral nutrition, are considered to be associated with candidemia in hospitalized heart failure patients. The identification of the route of infection in candidemia is difficult. In the current study, the positive blood and catheter cultures for Candida isolates and the similarity of the ITS region of ribosomal DNA sequence of Candida isolated from two patients confirmed the diagnosis of intravenous catheter-related candidemia.

Low In Vitro Antifungal Activity of Tavaborole against Yeasts and Molds from Onychomycosis.

The in vitro activity of tavaborole, an FDA-approved antifungal drug, was compared to that of four antifungal agents against 170 clinical fungal isolates originating from patients with onychomycosis. Tavaborole had low activity against all isolates compared to itraconazole, terbinafine, and fluconazole, the principal choices for treatment of onychomycosis. Thus, it appears that tavaborole is not a candidate for the treatment of onychomycosis due to Candida species, Aspergillus species, and dermatophytes.

Identification of Mucorales in patients with proven invasive mucormycosis by polymerase chain reaction in tissue samples.

BACKGROUND: Accurate diagnosis of mucormycosis, a life-threatening fungal infection, remains a challenge for physicians. OBJECTIVES: To identify the causative Mucorales in fresh clinical samples and formalin-fixed paraffin-embedded (FFPE) samples of patients with proven mucormycosis by molecular method. PATIENTS/METHODS: Fresh clinical samples of patients with proven mucormycosis according to the EORTC/MSG criteria admitted between 2015 and 2017 and histopathologically proven FFPE archives collected during 2004-2007 and 2015-2017 from Mazandaran University-affiliated hospitals of northern Iran were included. Seminested PCR targeting the 18S rDNA of Mucorales and ITS region was performed, and PCR products were then sequenced. RESULTS: While culture was positive only in 5 of 9 (56%) of fresh specimen cases, PCR was positive in all 9 (100%) histologically proven mucormycosis. Ten of 18 (56%) FFPE samples were PCR-positive. Overall, Mucorales PCR was positive in 19 of 27 (70%) samples. Mucorales species were Rhizopus arrhizus in 16 (84%) cases, R. arrhizus/Amylomyces rouxii in 2 (10.5%) cases and Rhizopus stolonifer in one case (5.5%). Among 27 mucormycosis cases, 25 (93%) cases were rhinocerebral, and 2 (7%) cases were disseminated. Diabetes mellitus (74%) and neutropaenia (63%) were the main risk factors. CONCLUSIONS: Seminested PCR targeting 18S rDNA region of Mucorales is useful for identification of the causative agents of mucormycosis.

Potent Activities of Luliconazole, Lanoconazole, and Eight Comparators against Molecularly Characterized Fusarium Species.

A collection of clinical (n = 47) and environmental (n = 79) Fusarium isolates were tested against 10 antifungal drugs, including 2 novel imidazoles. Luliconazole and lanoconazole demonstrated very low geometric mean MIC values of 0.005 and 0.013 µg/ml, respectively, compared with 0.51 µg/ml for micafungin, 0.85 µg/ml for efinaconazole, 1.12 µg/ml for natamycin, 1.18 µg/ml for anidulafungin, 1.31 µg/ml for voriconazole, 1.35 µg/ml for caspofungin, 1.9 µg/ml for amphotericin B, and 4.08 µg/ml for itraconazole. Results show that these drugs are potential candidates for (topical) treatment of skin and nail infections due to Fusarium species.

Invasive forms of Candida and Aspergillus in sputum samples of pulmonary tuberculosis patients attending the tuberculosis reference laboratory in Ghaemshahr, Northern Iran: An analysis of samples collected during the past 10years.

OBJECTIVE/BACKGROUND: Tuberculosis (TB) continues to be a major public health problem worldwide. This is especially true in Northern Iran, which has high TB prevalence. The chronic nature of this disease is further exacerbated if it is accompanied by fungal infection, which usually remains undiagnosed and thus untreated. Thus, mycotic infections add fatal dimensions to pulmonary TB. Our objective was to determine the prevalence of invasive forms of fungal elements in sputum samples collected from patients with pulmonary TB at a reference laboratory in Ghaemshahr, Northern Iran, during the past 10years. METHODS: In this retrospective study, sputum samples collected from 430 patients were examined. Pulmonary TB in patients was confirmed in our laboratory, and samples obtained during the period from March 2006 to February 2016 were analyzed. The sputum samples were subjected to biological (bacterial) staining (Ziehl-Neelsen and fluorochrome) and mycological investigation using KOH+ Calcofluor White (Sigma-Aldrich, India) by fluorescent microscopy and fungal culture on Sabouraud dextrose agar (Sigma-Aldrich, India) and CHROMagar (Paris, France). RESULTS: Invasive forms of fungal pathogens were observed as co-infection with Mycobacterium tuberculosis in 28/430 cases (6.51%). The frequency of Aspergillus, both branching and dichotomous infection, accounted for 3.72% (16/430): Aspergillus flavus, 1.63%; Aspergillus fumigatus, 1.16%; Aspergillus niger, 0.69%; and Aspergillus oryzae, 0.23%, respectively. Blastoconidia and pseudohyphae forms of yeast were observed as co-infection with M. tuberculosis in 2.79% (12/430) of the cases: Candida albicans, 1.86%; Candida krusei, 0.46%; and other Candida species, 0.46%, respectively. CONCLUSION: Northern Iran is a critical region in the TB world and multidrug-resistant TB is a serious problem in this region. Although it is believed that there exists a commensal relationship between fungus and TB infections, the invasive forms of fungal pathogens and their co-infection can be caused by increasing disability and failure of treatment. Diagnosis of secondary or co-existing fungal infections in TB is most important for reducing the mortality and morbidity of these patients.

Intravenous Catheter-Associated Candidemia due to Candida membranaefaciens: The First Iranian Case.

The incidence of candidemia due to the uncommon non-albicans Candida species appears to be increasing, and certain species such as Candida (C.) membranaefaciens have been reported in some clinical researches. Vascular catheters are considered the likely culprit for the sudden emergence of hospital-acquired candidemia. The identification of C. membranaefaciens can be problematic in clinical practice owing to its phenotypic resemblance to C. guilliermondii. We report the first case of C. membranaefaciens in Iran, which occurred in a 70-year-old woman, who had coronary artery bypass grafting (CABG). We isolated germ-tube negative yeast from both blood culture and central venous catheter (CVC) tip culture on brain-heart infusion agar, Sabouraud dextrose agar plates, and biphasic brain-heart infusion media bottle; it developed smooth, pink colonies on CHROMagar Candida. By using the polymerase chain reaction and sequencing of theinternal transcribed spacer region of rDNA, we identified C. membranaefaciens. After the removal of the CVC and initiation of Fluconazole treatment, the patient's condition gradually improved and she was discharged from the hospital. The early detection of organisms in the catheter, removal of the catheter, and treatment with anti-fungal antibiotics have an important role in controlling disease and preventing septicemia after CABG. As C. membranaefaciens is an opportunistic Candida species, both clinicians and microbiologists should be aware of the factors that confer fast diagnosis and appropriate treatment.

Isolation of different species of Candida in patients with vulvovaginal candidiasis from sari, iran.

BACKGROUND: Vulvovaginal Candidiasis (VVC) is a frequent, complex and cumbersome condition that can cause physical and psychological distress for the involved individual. Candida albicans was reported as the most common agent of VVC yet it seems that we are recently encountering changes in the pattern of Candida species in VVC. OBJECTIVES: In this study we assessed different species of Candida isolated from patients with VVC, residing in Sari, Iran. PATIENTS AND METHODS: Two hundred and thirty-four patients with vulvovaginitis were enrolled in this study. Samples were collected by a wet swab. Each vaginal swab was examined microscopically and processed for fungal culture. The identification of Candida species was done by morphological and physiological methods such as culture on CHROMagar Candida media and sugar assimilation test with the HiCandida identification kit (HiMedia, Mumbai, India). RESULTS: Out of 234 patients with vulvovaginitis, 66 (28.2%) patients showed VVC. Of these patients, 16 (24.2%) had recurrent VVC (RVVC). The age group of 20 - 29 year-olds had the highest frequency of VVC (48.5%). Erythema concomitant with itching (40.9%) was the most prevalent sign in VVC patients. Fifty-seven (86.4%) of the collected samples had positive results from both microscopic examination and culture. In total, 73 colonies of Candida spp. were isolated from 66 patients with VVC. The most common identified species of Candida were C. albicans (42.5%), C. glabrata (21.9%) and C. dubliniensis (16.4%). In patients with RVVC and patients without recurrence, C. albicans and non-albicans species of Candida were frequent species, respectively. CONCLUSIONS: The results of our study showed that non-albicans species of Candida are more frequent than C. albicans in patients with VVC. This result is in line with some recent studies indicating that non-albicans species of Candida must be considered in gynecology clinics due to the reported azole resistance in these species.

Detection of invasive aspergillosis in bone marrow transplant recipients using real-time PCR.

OBJECTIVE: The invasive aspergillosis (IA) is a serious opportunistic infection caused by various species of Aspergillus in immunocompromised individuals. Basically, rapid and early diagnosis prevents IA progression. In this study we performed a Real Time PCR/ Fluorescence Resonance Energy Transfer (FRET) for diagnosis of IA in hematologic malignancies and bone marrow transplant recipients. MATERIALS AND METHODS: Sixty two patients with hematologic malignancies and marrow transplant recipients were evaluated for IA in Sari and Tehran from 2009 to 2010. The primer and hybridization probe were designed to amplify the specific sequence of 18S rRNA genes using Light Cycler system and FRET. Galactomannan (GM) assay was performed on serums which obtained from selected patients using the Platelia Aspergillus kit. RESULTS: According to the criteria defined by the European Organization for Research and Treatment of Cancer and Mycoses Study Group (EORTC/MSG) for IA, 18 (29%) patients out of 62 patients were stratified into probable and possible groups. The female-to-male ratio was 1:2; the mean age of the patients was 36 years. The most common malignancies in these patients were acute lymphoblastic leukemia (38.9%). The minimum detection limit was 10 conidia (10(1) CFU/ml) equivalents (100 fg) per PCR reaction. GM assay was positive in 20.9% and real-time PCR probe set assay were positive in 17.7% patients who had clinical signs and host factor according to the mentioned criteria. CONCLUSION: Using the Real-Time PCR/FRET assay in whole blood specimens seems to be a promising method for diagnosis of IA, especially when used in combination with the GM detection test.