hossein zarrinfar

Candida parapsilosis is the second most common species causing infectious diseases and can lead to biofilm resistance. This study aims to adjust and synthesize a liposomal compound of Nigella sativa and evaluate its antifungal properties against C. parapsilosis isolates.

The liposomal formulation of N. sativa was optimized through the utilization of transmission elec- tron microscopy (TEM), particle size analysis, zeta potential measurement, and UV-visible spectrophotometry. Furthermore, an MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay was conducted on peripheral blood mononu- clear cells (PBMCs). The antifungal efficacy was evaluated in accordance with the M27-A3 guideline.

The minimum inhibitory concentrations (MICs) of N. sativa oil and the liposomal formulation on C. parapsilosis isolates ranged from 128 to 8 μg/mL and from 250 to 31.25 μg/mL, respectively. The MIC and MIC values of N. sativa oil and the liposomal formulation were 125, 187, and 32, 96 μg/mL, respectively. The viability percentage of cells treated with the liposomal formulation and free N. sativa oil was 91% and 85%, respectively.

The cytotoxicity of free N. sativa was significantly reduced when using nanoliposomes. The liposomal form of N. sativa showed greater antifungal properties compared to the free N. sativa extract against C. parapsilosis isolates.

Rhinosinusitis mucormycosis (RM) is an invasive opportunistic fungal infection, especially among COVID-19 patients. The current study aimed to assess the peripheral blood hematological disorders of COVID-19 patients-associated RM.
During ten month, in two COVID-19 centers in Mashhad, Iran, from June 2021 to March 2022, eighty-three patients suspected of COVID-19 with rhinosinusitis or rhino-orbital mucormycosis participated in this study. The hematological indices of these patients were measured by independent sample T‐test or Mann-Whitney test for quantitative data, and the qualitative variables were analyzed using Chi-square or Fisher’s exact test in SPSS version 20 at a significance level of 0.05.
Of the COVID-19 patients, 40 (48.2%) were affected by RM, and leukocytosis due to neutrophilia was observed in 30% of them. Leukocyte counts were normal in 10 (25%) patients, but 1 (2.5%) and 3 (7.5%) had leukopenia and lymphopenia, respectively. Leukocytosis plus lymphopenia was observed in 7 (17.5%) patients. Also, the synchronicity of leukopenia and lymphopenia was seen in 5 (12.5%) patients. Leukopenia, lymphopenia, and neutropenia have occurred concurrently in 2 (5%) patients. The complete blood count (CBC) showed that RBCs, hemoglobin (Hb), hematocrit (HCT), MCH, MCHC, platelet (PLT), and lymphocytes decreased while neutrophils increased.
Among the hematological parameters, leukocytosis due to neutrophilia and reduction in Hb, HCT, and PLT are more dominant factors in COVID-19 patients-associated RM.

 Candida parapsilosis complex, as an opportunistic pathogen, can cause various infections, especially in immunocompromised patients.

 This study investigated a novel thymoquinone-liposomal nanoparticle to increase the stability of thymoquinone with antifungal effects against C. parapsilosis isolates.

 The thymoquinone was encapsulated in liposomal nanoparticles using a thin-film hydration technique and then analyzed by transmission electron microscopy (TEM), particle size, zeta potential, and UV-visible spectrophotometer. Also, the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay was used on peripheral blood mononuclear cells (PBMCs) for cell metabolic activity. The antifungal activity of thymoquinone-liposomal nanoparticle against 15 clinical isolates of C. parapsilosis and the reference strain was examined based on the M27-A3 guideline.

 The synthesized thymoquinone-liposomal nanoparticle was approved by the TEM, particle size, zeta potential, and UV-Vis. The thymoquinone-liposomal nanoparticle showed no toxic effect on PBMCs following the MTT assay. The minimum inhibitory concentration (MIC) range of free thymoquinone and liposomal formulations with inhibitory effects on Candida isolates was 50 to 6.25 and 150 to 18.75 µg/mL, respectively. The MIC50 and MIC90 values of free thymoquinone and thymoquinone-liposomal nanoparticle were 25 and 50, and 75 and 150 µg/mL.

 The synthesized thymoquinone-liposomal nanoparticle shows significant antifungal activity against C. parapsilosis isolates compared to free thymoquinone. However, because of its hydrophilicity and hydrophobicity, biocompatibility, particle size, non-toxic effect, and higher cell viability, the thymoquinone-liposomal nanoparticle is a more effective method for treating fungal infections.

Helicobacter pylori is a gram-negative curved bacillus that assumes a significant role in colon cancer and children´s diseases. This study aimed to examine the association between H. pylori infection with colon cancer and children´s diseases in order to achieve a a comprehensive understanding of these associations and for future works.

Three main databases (i.e., Cochrane Central Register of Controlled Trials [CENTRAL], Scopus, and MEDLINE) were systematically searched by two reviewers from their inception date to 2022 in order to determine the association between the H. pylori infection with the colon cancer and children’s diseases.

The findings of two meta-analyses were similar regarding the positive association between the risks of colorectal neoplasm (pooled OR=0.18; 95% CI of 0.99–1.40; P>0.05) and colon neoplasia (pooled OR=0.41; 95% confidence interval 1.24–1.60; P=0.000). H. pylori was associated with an increased risk of colorectal adenoma, adenocarcinoma, and advanced adenoma. Also H. pylori infection was correlated with a high risk of iron deficiency anemia (IDA), otitis media with effusion (OME), HenochSchonlein purpura, and growth disorders in children.

In sum, the H. pylori infection may have been associated with an increased risk of colorectal cancer and children’s diseases.

Aspergillus species are found as opportunistic agents to cause a wide variety of clinical manifestations. Regarding the drug resistance emergence against Aspergillus species, new aspects of using nanoparticles (NPs) as antifungal agents are considerable. This study takes a new approach to biosynthesized NPs of zinc oxide, copper oxide, cerium oxide, silver, gold, and selenium influence on the clinical isolates of Aspergillus species.

The antifungal activities of six NPs were examined against a total of 12 clinical isolates of Aspergillus species, including A. flavus (n=4), A. welwitschiae (n= 4), and A. fumigatus (n=4) based on the M38-A2 guideline.

According to minimum inhibitory concentration (MIC) values, NPs of ZnO, Ag, Au, and Se showed a significant antifungal effect. CuO-NPs and CeO2-NPs didn’t show an inhibitory effect against Aspergillus isolates. The MIC ranges of ZnO-NPs, Ag-NPs, Au-NPs, and Se-NPs were 128-512, 26-53, 21-85, and 6-26 µg⁄mL for A. fumigatus; and 512->512, 26-53, 85, and 1-13 µg⁄mL for A. welwitschiae, respectively. In addition, the MIC ranges of Ag-NPs and Se-NPs were 26-53 and 106-425 µg⁄mL for A. flavus, respectively. However, A. flavus were not inhibited by NPs of ZnO and Au.

Among the examined NPs, ZnO, Ag, Au, and Se showed a significant effect against Aspergillus isolates except for CuO and CeO2. However, Ag-NPs seemed to be the most effective nanoparticle against the Aspergillus species. Compared to other Aspergillus species, A. flavus was not inhibited by NPs of ZnO and Au.

Green synthesis as a new method of synthesis of nanoparticles with a simple, biocompatible, safe, and economical approach can be an alternative to chemical and physical processes. Fungi can convert some toxic ions into less harmful forms, including nanoparticles. Nanoparticles with a size of 1 to 100 nanometers have unique quantum properties. Today, the problems of drug resistance have been seen in different species of fungi. Selenium nanoparticles (SeNPs) are substances that have been reported to have antifungal properties. The present study aimed to investigate the antifungal effect of biosynthesized SeNPs using Aspergillus fumigatus.

For this purpose, SeNPs were biosynthesized with a specific concentration using A. fumigatus. The presence of nanoparticles was confirmed by various methods, including UV-Vis, FT-IR, FE-SEM, EDX, XRD, DLS, and Zeta potential. Then, susceptibility determination based on the Minimum Inhibitory Concentration (MIC) testwas performed on standard fungal strains treated with SeNPs.

After confirming the results of nanoparticle biosynthesis, the MICs for Itraconazole and Amphotericin B against the standard fungal strains were 8 and 4 μg/mL respectively. In comparison, MIC values for SeNPs-treated samples were reduced to 1 μg/mL and below.

Due to the increasing resistance of opportunistic fungi to target antifungal drugs, the use of biosafety SeNPs even at low concentrations can have favorable inhibitory effects on the growth of fungal pathogens

Aspergillus rhinosinusitis is a potentially lethal complication in patients with underlying immunodeficiencies. Critical laboratory findings, such as lymphopenia and leukocytosis, were highly reported in coronavirus disease 2019 (COVID-19) patients. Based on the correlation between COVID-19 and fungal infections, this study was designed to evaluate the hematologic parameters in COVID-19 patients associated with Aspergillus rhinosinusitis in northeaster Iran. During eight months and among 80 COVID-19 patients suspected of fungal rhinosinusitis, in two tertiary referral hospitals of Mashhad, hematological parameters, such as white blood cell (WBC) count, of 14 patients affected to COVID-19 with Aspergillus rhinosinusitis were precisely evaluated to check leukopenia and leukocytosis. The patients showed a range of 42 to 67 years old and a median age of 59. Of the 14 patients, 8 (57.4%) had diabetes mellitus, 9 (64.2%) died, and two patients has normal leukocyte count. The three, two, and one patients showed leukocytosis, lymphopenia, and leukopenia, respectively. The combination of leukocytosis and lymphopenia was significant in four patients. However, leukopenia and lymphopenia were observed in just one subject. Moreover, leukopenia, lymphopenia, and neutropenia were detected together in one case.Aspergillus rhinosinusitis had high mortality among COVID-19 patients. Moreover, the high rate of diabetes mellitus was a severe predisposing factor for COVID-19-associated Aspergillus rhinosinusitis. Leukocytosis (neutrophilia) and lymphopenia were the most common hematological abnormalities among COVID-19 patients with Aspergillus rhinosinusitis.

Resistance to antifungal drugs is increasing among Candida isolates from patients with vulvovaginal candidiasis (VVC). Lack of correct diagnosis of Candida causing VVC and the experimental use of antifungal drugs are the main causes of this resistance. This study aimed to determine the susceptibility of antifungal drugs against Candida species isolated from VVC in Northeastern Iran.

Among women suspected of VVC, 189 vaginal dischargespecimens were evaluated. Candida isolates detected by polymerase chain reactionrestriction fragment length polymorphism were examined by standard antifungal diskdiffusion susceptibility testing method for voriconazole, itraconazole, fluconazole, andketoconazole. The susceptibility pattern of these antifungals was reported as sensitive,susceptible dose-dependent, and resistant. The results were evaluated by SPSS software and analyzed by Pearson chi-squared test.

Among the vaginal specimens, 108 out of 189 Candida isolates were identifiedas C. albicans, C. glabrata, C. kefyr, C. parapsilosis, and C. tropicalis. The susceptibilityrates of Candida isolates to voriconazole, ketoconazole, itraconazole, and fluconazolewere 92.6%, 90.7%, 68.5%, and 63.9%, respectively. Moreover, the resistance rates tofluconazole, itraconazole, ketoconazole, and itraconazole were 15.7%, 8.3%, 1.9%, and1.9%, respectively. The C. glabrata and C. albicans isolates were resistant to antifungaldiscs among 93% and 20% of the specimens, respectively.

The C. glabrata and C. albicans species showed the highest resistance toantifungal drugs. Furthermore, Candida isolates showed the highest sensitivity tovoriconazole and ketoconazole and the lowest sensitivity to fluconazole.

Candida vulvovaginitis is one of the most common vaginal infections in women and in women with diabetes. Although C. albicans has been reported to be the most common cause of this infection, but there have been recent changes in the pattern of Candida causing Candida vulvovaginitis. This study was performed with aim to investigate the frequency of dominant Candida isolates in patients with Candida vulvovaginitis in Torbat-e Jam and its relationship with diabetes.

In this cross-sectional study, 428 women with suspected Candida vulvovaginitis referred to Torbat-e-Jam medical centers were assessed in 2019. Two specimens of vaginal secretions were taken from each patient. The first specimen was cultured on sabouraud dextrose agar (SDA) medium, and the second one was directly examined using wet mount microscopy. The Candida colonies were passaged on CHROMagar Candida medium to identify the species. Demographic and diabetes characteristics of the patients were evaluated. Data were analyzed by SPSS software (version 16) and Chi-square test. p<0.05 was considered statistically significant.

309 positive yeast cultures were obtained from the patients with Candida vulvovaginitis. Identified Candida species included: 145 (46.9%) C. albicans, 53 (17.2%) C. glabrata, 45 (14.5%) C. parapsilosis, 33 (10.7%) C. krusei, and 33 (10.7%) C. tropicalis. Forty-one specimens (13.3%) also had two types of Candida species. Forty-one patients (13.3%) affected to Candida vulvovaginitis also had diabetes that no significant relationship was observed.

The frequency of non-albicans Candida is relatively higher than C. albicans in patients with Candida vulvovaginitis in Torbat-e Jam, and C. glabrata is the most common among non-albicans species. The incidence of Candida vulvovaginitis was not significant in patients with diabetes. C. parapsilosis was most common among non-albicans species in diabetics patients.

Keratinophilic fungi play an important role in the decomposition of keratinous substances in nature. This capacity induces dermatomycosis in both humans and livestock. The soil of livestock stables can be a reservoir of keratinophilic fungi. Therefore, the present study was conducted to isolate and identify keratinophilic fungi in the soil of the livestock stables located in Qayen, South Khorasan Province, Iran.

This study was conducted on 62 soil samples collected from livestock stables. The samples were cultured by means of hair bait technique (HBT). The identification of the isolates was performed based on their morphological characteristics and then confirmed by polymerase chain reaction and sequencing of the ITS regions of ribosomal DNA.

A total of 118 isolates of 7 species from 5 genera were identified. Aphanoascus verrucosu (n=70, 59.36%) was detected as the dominant keratinophilic fungus, followed by Arthroderma quadrifidum (n=13, 11.01%), Aphanoascus terreus (n=12, 10.16%), Acremonium (n=12, 10.16%), Arthroderma gertleri (n=5, 4.23%), Fusarium equiseti (n=3, 2.54%), and Uncinocarpus reesii (n=3, 2.54%).

Different keratinophilic fungi were isolated from the soil of livestock stables; however, Aphanoascus verrucosu was found to be the dominant species.

Phaeohyphomycosis is a rare opportunistic fungal infection that can be caused by dematiaceous fungi. The clinical manifestations of this disease are highly diverse and include cutaneous, subcutaneous, and systemic forms. However, the mortality rate of this infection depends on the immune status of patients. Although infections caused by black fungi have been on an increasing trend during the past decade, infections with dematiaceous fungi have rarely been reported in the Middle East. However, some fungal species are associated with a disseminated disease without the known clinical and pathologic risk factors. The analysis of DNA sequence is a highly effective method for differentiating various species among some genera, due to the variable cultural and morphological characteristics of DNA. Despite the presence of therapeutic methods, there are numerous reports regarding the high mortality rate of this infection. Published studies indicate no specific risk factors for phaeohyphomycosis and report the incidence of this disease in immunocompetent individuals. Moreover, selective treatment for these rare infections has not even been defined in clinical studies. However, the clinical syndromes associated with dematiaceous fungi should be considered in the Middle East, Iran.

Dermatophytes are a group of fungi specialized in invading humans and other vertebrate keratinized tissues. These fungi cause a variety of skin, nail, and hair disorders, called dermatophytosis (tinea). In some cases, drug resistance to antifungals necessitates special treatment. Among the antifungal agents, sertaconazole (i.e., a third-generation imidazole) has a broad-spectrum against dermatophyte species. Regarding this, the present study was conducted to investigate the antifungal susceptibility of dermatophytes obtained from patients with dermatophytosis in Mashhad located in northeastern Iran.

A total of 75 clinical dermatophyte isolates, including Trichophyton mentagrophytes (n=21), T. interdigital (n=18), T. tonsurans (n=16), Epidermophyton floccosum (n=11), Microsporum canis (n=5), Nannizzia fulvum (n=2), T. benhamiae (n=1), and T. verrucosum (n=1), were evaluated against five antifungal agents of sertaconazole, itraconazole, clotrimazole, terbinafine, and griseofulvin based on the CLSI M38-A2 guideline.

According to the results, the minimum inhibitory concentration (MIC) ranges of sertaconazole, terbinafine, griseofulvin, itraconazole, and clotrimazole were estimated at 0.125-16, 0.002-1, 0.5-4, 0.031-4, and 0.016-4 μg/ml, respectively, for dermatophyte species. In addition, the geometric mean (GM) values of the MIC of sertaconazole, terbinafine, griseofulvin, itraconazole, and clotrimazole were obtained as 3.39, 1, 1.44, 1.52, and 1.93, respectively.

Among the tested antifungals, terbinafine and griseofulvin were the most effective agents against dermatophyte isolates. However, sertaconazole, a third-generation imidazole, did not show any significant effect. Furthermore, M. canis and E. floccosum showed the best response to the antifungal agents.

Dermatophytes as the causative agents of dermatophytosis (ringworm) are widely spread around the world. Accurate identification of dermatophytes in one area can be particularly important for epidemiological studies. Regarding this, the aim of the present study was to describe the species spectrum of dermatophytes, isolated from patients in Mashhad city, Iran, using the molecular-based method.

This study was conducted on 79 dermatophyte isolates obtained from the human skin, hair, and nail specimens. Species identification was performed by the polymerase chain reaction-restriction fragment length polymorphism analysis of ribosomal DNA internal transcribed spacer regions using MvaI restriction enzyme.

The identified species included Trichophyton mentagrophytes/T. interdigitale species complex (n=37, 46.8%), Epidermophyton floccosum (n=12, 15.2%), T. rubrum (n=8, 10.1%), Microsporum canis (n=8, 10.1%), T. violaceum (n=5, 6.3%), T. tonsurans (n=4, 5.1%), Nannizzia gypsea (n=3, 3.8%), T. benhamiae (n=1, 1.3%), and T. verrucosum (n=1, 1.3%). The clinical forms of infection were tinea corporis (n=26, 32.8%), tinea cruris (n=22, 27.8%), tinea capitis (n=10, 12.6%), tinea unguium (n=7, 9%), tinea manuum (n=6, 8%), tinea pedis (n=5, 6.3%), and tinea faciei (n=3, 3.5%).

As the findings indicated, T. mentagrophytes/T. interdigitale species complex had the highest prevalence, and T. benhamiae appeared to be a new emerging agent of dermatophytosis in Mashhad, northeastern Iran.

Dermatophytes are a group of fungi that attack keratinous tissues of the skin, hair, and nail in humans and animals, and cause infections called dermatophytosis (tinea). Since identification of pathogenic fungi at the species level is essential for the detection of the source, control and prevention, and identifying epidemiology of infection, it is necessary to use specific and sensitive diagnostic methods to identify the causes of dermatophytosis. The clinical samples (skin, nail, and hair) of patients with dermatophytosis in Mashhad City, Iran, were cultured in Mycosyl Agar culture media, and the DNA of obtained dermatophyte colonies were extracted by specific kit. The internal transcribed spacer (ITS) gene was amplified and sequenced by ITS1, ITS4 primers. Finally, the sequencing results were analyzed using SeqMan software, and were compared with the data of the global genebank. In this study, 80 dermatophyte isolates were sequenced, which included 9 dermatophyte species as 23 (28.8%) Trichophyton (T.) interdigital, 18 (22.5%) T. tunsorans, 10 (12.5%) Epidermophyton fluccosum, 10 (12.5%) of T. mentagrophytes, 8 (10%) Microsporum canis, 4 (5%) T. rubrum, 4 (5%) T. benhamiae, 2 (2.5%) Nannizzia (N.) fulvum, 1 (1.2%) N. persicolor. According to report the rare species of dermatophytes in this study, the use of molecular methods such as sequencing of the ITS gene can determine the diversity of dermatophytes in a region more precisely than morphological methods.

Candidiasis is referred to a group of superficial and deep-tissue fungal infections often caused by Candida albicans. The superficial infections affect the oral, oropharynx, esophagus, and vaginal mucosa. The treatment of choice for these infections is the use of azoles, such as fluconazole. However, the increased use of these antifungal agents has led to the emergence of azole-resistant isolates of C. albicans. Different mechanisms have been suggested for the development of drug resistance, such as mutations in the encoding gene ERG11. Mutations in ERG11 result in changes in the ERG11p spatial construction and reduce the affinity between the protein and azole. This study aimed to determine the susceptibility profile of C. albicans clinical isolates to fluconazole using microdilution method. The present research was also targeted toward the detection of mutations that might be related to fluconazole resistance by the amplification and sequencing of ERG11 gene. This study was conducted on a total of 216 clinical isolates obtained from Mashhad, Isfahan, and Tehran cities in Iran, during 2016-2018. The clinical isolates were identified using molecular techniques. Furthermore, minimum inhibitory concentration (MICs) was determined according to the clinical and laboratory standards institute M27-A3 and M27-S4 documents. The concentration range for fluconazole was obtained as 0.063-64 μg/ml. In the resistant strains, ERG11 genes were amplified by specific primers. Subsequently, cycle sequencing reactions were performed on purified polymerase chain reaction (PCR) products in forward and reverse directions. Finally, the results were analyzed by MEGA (version 7) and Gene Runner software (version 6.5.30). Out of 216 strains, 100 (46.3%) species were identified as C. albicans. The MIC values for fluconazole had a range of 0.125-16 μg/ml with the MIC50 and MIC90 values of 0.5 and 1 μg/ml, respectively. Totally, 41 nucleotide changes were detected among 4 resistant isolates. In this regard, 4 out of 41 mutations in codons caused changes in ERG11p; however, these mutations did not lead to fluconazole resistance. Fluconazole resistance among clinical isolates is not merely due to the changes in ERG11p. This resistance may be also related to some other mechanisms, such as the prevention of the intracellular accumulation of the antifungal agent and alteration of the target enzyme to diminish drug binding.

Background and Candida-associated denture stomatitis is one of the most common forms of oral candidiasis among denture wearers. Regarding this, the aim of the present study was to evaluate the antifungal effects of home-generated ozonated water on the adhesion of the C. albicans attached to the surface of the denture base acrylic resins. For the purpose of the study, different concentrations of C. albicans were added to the tubes containing acrylic resin blocks, and then incubated for 2 h at 35°C. The samples were assigned into three groups, each of which contained 42 samples, including normal saline (NS) solution as the negative control, nystatin (N) solution as the positive control, and ozonated water as the test group. The samples were washed and placed in an ultrasonic bath. Subsequently, the saline solution was cultured on Sabouraud dextrose agar. The concentrations of Candida were evaluated during the contact times. The test group (i.e., ozonated water) with 114 colony-forming units (CFU) showed a significant reduction of Candida colonies, compared to the NS group with 2,172 CFU. The 120- and 1-minute incubation with ozonated water showed the highest and lowest effects on the viability of Candida adhered to the acrylic resin, respectively. Based on the findings, home-generated ozonated water can be applied to remove the Candida attached to the surface of the denture plates

Background and PurposeVulvovaginal candidiasis (VVC) is a common problem in women. The purpose of this study was to identify of Candida species isolated from vulvovaginitis woman suffering vulvovaginitis refered to Ghaem Hospital, Mashhad, Iran, by use of MALDI-TOF mass spectrometry.
Materials and MethodsThe 65 clinical samples isolated from Vulvovaginitis women were collected in Ghaem Hospital. All specimens were identified using phenotypic techniques such as microscopy and culture on Sabouraud dextrose agar and corn meal agar medium,Then, All isolates were detected and were processed for MALDI TOF MS identification.
Results Of the 65 isolates analyzed, 61 (93.8%) were recognised by MALDI-TOF mass spectrometry and for four isolates (6.1%) only not relabile identifications were achieved. In this study, the most frequently isolated species were Candida albicans (58.5%), followed by Candida tropicalis (16.9%), Candida glabrata (7.7%), Candida parapsilosis (7.7%) and Candida guillermondii (3.1%).
Conclusionpresented results demonstrate that the MALDI TOF mass spectrometry is a fast and reliable technique, and has the potential to replace conventional phenotypic identification of Candida species and other yeast strains routinely isolated in clinical microbiology laboratories.

Background and Rhinosinusitis is a common disorder, influencing approximately 20% of the population at some time of their lives. It was recognized and reported with expanding recurrence over the past two decades worldwide. Undoubtedly, correct diagnosis of fungi in patients with fungal rhinosinusitis affects the treatment planning and prognosis of the patients. Identification of the causative agents using the standard mycological procedures remains difficult and time-consuming.
Based on clinical and radiological parameters, 106 patients suspected of fungal rhinosinusitis were investigated in this cross-sectional prospective study from April 2012 to March 2016 at an otorhinolaryngology department. In this study, internal transcribed spacer (ITS) and calmodulin (CaM) sequencing were respectively validated as reliable techniques for the identification of Mucorales and Aspergillus to species level (both agents of fungal rhinosinusitis).
Of these, 63 (59.4%) patients were suspected of allergic fungal rhinosinusitis (AFRS), 40 (37.7%) patients suspected of acute invasive fungal rhinosinusitis (AIFRS), and 3 (2.8%) patients suspected of fungus ball. In patients suspected of AFRS, AIFRS, and fungus ball only 7, 29, and 1 had positive fungal culture, respectively. After ITS and CaM sequencing, Aspergillus flavus was the most common species isolated from non-invasive forms, and A. flavus and Rhizopus oryzae were more frequently isolated from invasive forms.
Aspergillus flavus is the most common agent of fungal rhinosinusitis in Iran, unlike most other reports from throughout the world stating that A. fumigatus is the most frequent causative agent of this disease.

Malassezia yeast can become pathogenic under certain conditions in humans. Malassezia has various species which can cause a number of skin diseases, such as pityriasis versicolor (tinea versicolor), seborrheic dermatitis and folliculitis and even systemic infections. In the present study, Malassezia species isolated from the patients with pityriasis versicolor were identified by PCR-PFLP molecular method.
Material and In this study, the scraping specimens of the trunk and scalp of the patients with pityriasis versicolor were cultured on Dixon agar medium. Finally, one-hundred Malassezia colonies were obtained. The genomic DNA was extracted by phenol–chloroform method and then was studied by use of PCR-PFLP molecular method. D1-D2 segment in the area of 26srDNA of ITS gene was proliferated by specific primers and then the PCR products were exposed to CfoI restrictive enzyme.
Among 100 Malassezia colonies, the most common Malassezia species were; M. globosa (44%), M. globosa/M. restricta (27%), M. restricta (11%), M. sympodialis (7%), M. sympodialis/M. restricta (4%), M. globosa/M. restricta/M. furfur (1%), M. sympodialis/M. restricta/ M. globosa (1%) and unknown species (5%).
The dominant species isolated from patients with pityriasis versicolor were M. globosa, M. restricta and M. sympodialis, respectively. Thirty-three percent of the specimens had more than one Malassezia species. Therefore, rapid PCR-RFLP method is recommended for identification of Malassezia species in epidemiological studies and production of more effective medicines.

Vulvovaginal candidiasis (VVC) is one of the most common vaginal infection in women of childbearing age. In some cases, due to the intervention of resistant strains of Candida albicans and non-albicans species, recurrent vulvovaginal candidiasis (RVVC) occurs. This study was performed with aim to identify the Candida isolated from patients with vulvovaginal candidiasis by PCR-RFLP method.
This descriptive study was performed on 189 vaginal discharge obtained from women with clinical presentation of VVC referred to Ghaem Hospital, Mashhad University of Medical Sciences during 2015-2016. The vaginal discharge was examined by direct examination and culture on Sabouraud's dextrose agar medium for definitive diagnosis. The genomic DNA of Candida colonies have grown in culture were extracted by boiling lysis method. The Candida isolates were identified by PCR-RFLP technique. The results were analyzed by SPSS software (version 16) and using Pearson and Chi-square test. P
Among 189 sampled patients, 108 positive Candida cultures were acceptable. The Candida species which were identified included C. albicans 84 (77.8%), C. glabrata 10 (9.3%), C. kefyr 4 (3.7%), C. parapsilosis 1 (0.9%) and C. tropicalis 1 (0.9%). About 8 (7.4%) of the specimens contained two species of Candida. RVVC form was observed in 5 (4.6%) of the patients.
The most common cause of VVC was C. albicans; and C. glabrata had the highest frequency among the non-albicans species. Most of patients had non- RVVC.

The use of immune suppressive drugs for organ transplant recipients predisposes them to opportunistic infections, especially by fungal agents. Pneumocystis jiroveci, as an opportunistic pathogen, endangers the patients’ life in those with immune system disorders. Early detection of latent Pneumocystis infection in susceptible patients may help choose the optimal treatment for these patients.. The aim of this study was to identify and determine the colonization of latent P. jiroveci infection among lung transplant recipients..Patients and This cross-sectional descriptive study was conducted on lung transplant recipients. Bronchoalveolar lavage (BAL) specimens were collected from 32 patients undergoing bronchoscopy. The samples were aseptically homogenized by 10 mM dithiothreitol, and their DNA was extracted. The mtLSUrRNA gene of P. jiroveci was amplified using nested PCR in two stages. Nested PCR was performed using external primers of pAZ-102-E and pAZ102-H followed by using the PCR product of the first stage and internal primers of pAZ-102-E and pAZ102-L2.. The genome of P. jiroveci was revealed by a 346 bp PCR product in the initial amplification and a 120 bp product in the nested PCR. The results showed that seven BAL specimens (21.9%) from lung transplant recipients were positive for P. jiroveci.. In molecular epidemiology studies, nested PCR has higher sensitivity than PCR. Results of this study support the colonization of P. jiroveci in patients receiving lung transplantation. Patients who are carriers of P. jiroveci are at a higher risk of P. jiroveci pneumonia..

Toxoplasma gondii is an opportunistic parasitic organism causing infection in many mammals, including immunosuppressed patients. Toxoplasmosis as an opportunistic infection is highly prevalent among patients receiving a kidney transplant.. The purpose of this study was to identify and determine the prevalence of Toxoplasma gondii in clinical samples collected from patients receiving renal transplants..Patients and A total of 50 blood samples and 40 lung lavage samples from transplanted patients admitted to the infectious wards and the patients undergoing bronchoscopy were collected. The B1 Gene of Toxoplasma gondii was amplified using PCR of the blood and bronoalveolar lavage BAL samples, and IgG and IgM antibodies against Toxoplasma were detected in serum samples using ELISA.. Our results indicated that anti-toxoplasma specific IgG and IgM antibodies were prevalent among transplant recipients with values of 54% and 4% respectively. PCR was performed to detect Toxoplasma gondii in 3 blood and lavage samples (3.3%) with 100% sensitivity and 97.9% specificity.. Toxoplasma gondii pulmonary infection is measured along with brain toxoplasmosis in patients receiving a kidney transplant. After serological methods, PCR is the second useful method for Toxoplasma gondii screening. Proper prophylaxis before and after receiving a kidney transplant together with Toxoplasma gondii screening of donor and transplant is recommended..

Malassezia species as skin microflora of humans and other warm-blooded vertebrates are the lipophilic yeasts associated with various human diseases, especially pityriasis versicolor (PV).. The objective of this study was to identify the Malassezia species of scraped skin of PV patients in Yazd, Iran, using morphological, biochemical and physiological methods. We also compared the obtained results of PV patients with normal healthy volunteers.. A total of 200 persons, including 100 patients (with skin lesion) referred to Yazd Central Laboratory and 100 healthy volunteers as controls, were evaluated for Malassezia infection by morphological and biochemical methods.. The most commonly isolated species from PV lesions, were M. globosa (38.3%), M. furfur (29.4%), M. sympodialis (14.9%), M. pachydermatis (9.6%) and M. slooffiae (5.3%). Also the most commonly isolated species from healthy skins were M. furfur (37.2%), M. globosa (25.6%), M. sympodialis (16.3%), M. pachydermatis (13.9%) and M. slooffiae (4.6%). Totally M. globosa and M. furfur were the most frequented isolated.. Highest prevalence of PV in our study was observed in the 20 – 39 years old group, suggesting that the peak of the infection is coincided with ages and increasing sebum production in the highest level. M. globosa was the most commonly isolated Malassezia species of the patients group and M. furfur is the most common isolated species obtained from normal individuals skin samples. We couldn’t find any significant differences between groups. The rate of isolated Malassezia species from patients was higher than normal individuals..

Pneumocystis jirovecii causes Pneumocystis pneumonia (PCP) in immunocompromised patients with a high rate of morbidity and mortality. Colonization with this fungus may stimulate pulmonary inflammation or lead to PCP in susceptible patients. The epidemiology of this infection and routs of its transmission has poorly studied in Iran. We examined Pneumosystis colonization in patients with various lung underlying diseases. Bronchoalveolar lavage (BAL) fluids of 458 patients with different underlying diseases or pulmonary signs were collected between August 2010 and January 2012. Patients were divided into four groups: transplant recipients, malignant patients, immunosuppressive drug recipients and patients with other different lung diseases. A sensitive nested-PCR method targeted 18S ribosomal RNA gene was used for investigating P. jirovecii in the specimens. P. jirovecii DNA was detected in 57 out of 458 (12.5%) BAL samples by nested-PCR. Colonization rate in malignant patients, transplant recipients, immunosuppressive therapy recipients and patients with other various lung diseases was 21.7%, 20.3%, 12.7% and 7.3%, respectively. The enzyme BanI cuts all PCR products producing fragments with the size of 228 and 104 base pair. This finding as well as sequencing of four random positive samples validated and reconfirmed the PCR results. P. jirovecii cysts were found in 5 out of 57 PCR positive samples. A significant number of patients with pulmonary diseases were colonized by P. jirovecii that can develop to PCP in these patients or they may transmit the fungus to other susceptible patients.