Green Synthesis of Silver Nanoparticles by Fusarium oxysporum and its Function Against Aspergillus and Fusarium Fungi.

(NPs) can be produced by various methods such as physical and chemical processes. However, environmentally friendly ways are increasingly requested. In this research, (Ag-NPs) were produced by Fusarium oxysporum, and its antifungal effect on Aspergillus and Fusarium was investigated. Nanoparticles were produced by silver nitrate salt and Fusarium oxysporum native to Isfahan city. In order to optimize the synthesis conditions, optimization of some factors such as volume, concentration, time, temperature, and pH of the extract was performed. The structural and physical properties of NPs were determined by spectrophotometer, XRD, FTIR FESEM, SEM, and TEM microscopy. For the study of the inhibitory effect of NPs on Fusarium and Aspergillus growth, the fungi were cultured in media containing various concentrations of NPs from 50 to 1500 ppm. Then, the colony diameter was measured for over 10 days and the growth inhibition percentage was estimated. For statistical analysis, the 600 Mann-Whitney tests have been applied.The NPs were produced after mixing the powdered fungal mass and silver nitrate salt in optimum conditions which were 2 mM of salt, triple fungal mass volume proportion relative to the salt, pH of 9, and temperature of 28 °C. The existence of a peak at 420 nm in FTIR was due to nanoparticle production. Based on the XRD, the synthesized NPs had suitable properties similar to the standard NPs reported in the studies. Images from TEM, SEM, and FESEM microscopes displayed uniform NPs in variable sizes between 25 and 100 nm. According to the results, the maximum growth inhibition percentage of Ag-NPs on Fusarium was approximately 60% at 1500 ppm, and 88% on Aspergillus at 800 ppm. Biosynthesized Ag-NPs with Fusarium oxysporum have desirable structural traits and can inhibit the growth of Fusarium and Aspergillus at significant levels.

Periplaneta americana (Blattodea: Blattidae) fungal pathogens in hospital sewer systems: molecular and phylogenetic approaches.

Cockroaches are known as mechanical vectors of some pathogens that can infect humans. The present study aims to rapidly identify Periplaneta americana fungal pathogens from sewer systems of public hospitals in Esfahan using the polymerase chain reaction (PCR) technique. A total of 55 P. americana cockroaches were randomly collected by direct trapping from sewer systems of seven hospitals and screened for fungal infectious agents using standard morphological methods and the PCR sequencing. From the American cockroach, we isolated 62 yeasts and 31 molds from the surface, hemocoel, and digestive tract of P. americana. Based on DNA sequence comparisons and other taxonomic characteristics, they were identified as more than four species of yeast and four species of mold. Yeast species including Pichia kudriavzevii, Candida glabrata, Pichia kluyveri, and Candida viswanathii, and molds such as Aspergillus niger, Penicillium italicum, Mucor plumbeus, and Rhizopus oryzae were isolated repeatedly from the surface, hemocoel, and digestive tract of P. americana. Our results show that the use of a combination of morphological, molecular techniques, and phylogenetic analysis can lead to the identification of pathogenic fungal agents in American cockroaches and also knowledge of fungal pathogens-arthropod host relationships.

Isolation of Bacillus amyloliquefaciens M13-RW0 from Soil and Evaluation of its Antifungal Activity.

BACKGROUND: The frequency of observed invasive Aspergillosis has increased in recent years. Infection with other molds happens but does not lead to a large proportion of invasive infections. The present study aims to isolate Bacillus amyloliquefaciens M13-RW0 from soil and evaluate its antifungal effects against some saprophytic fungi, such as Aspergillus niger, Aspergillus flavus, and Mucor hiemalis. MATERIALS AND METHODS: In this research, a total of 150 samples (from the soil, air, and surfaces) were prepared from different parts of Isfahan, Iran. Isolation and purification of growing bacteria were conducted using the nutrient agar medium. The inhibitory effects of 100 isolated bacteria were evaluated against the growth of A. niger, A. flavus, and M. hiemalis, 4 bacteria were isolated with inhibitory effects against the selected fungi, and consequently, one of the bacteria isolated from the soil samples was found to show the highest inhibition of fungal growth. Quantitative evaluation of the growth inhibitory effect was performed using linear culturing of fungal suspension (104 spore/ml) at distances of 5, 10, 15, 20, 25, and 30mm from bacterial isolate (0.5 McFarland) on Sabouraud Dextrose Agar (SDA) medium. Results were checked 24, 48, 72, and 96 hours later. The bacterial isolate with the most inhibitory effect was identified by phenotypic and molecular tests. RESULTS: According to the results, among the 4 inhibitory bacterial isolates, Bacillus amyloliquefaciens strain M13-RW01, isolated from the soil samples, was identified as the bacterium with the most significant potential antifungal activity. The strong inhibitory effect was revealed after 48 hours for all distances of 15mm and more between the fungi and the bacterium. CONCLUSION: The identified bacterium could not only be considered an inhibitor bacterium against saprophytic fungi but could also be put forward to help produce new antifungal drugs for controlling fungal diseases.

Antifungal Effect of Carrot Carotenoids on Candida Species.

BACKGROUND: Candidiasis is a serious problem in women's health that is caused by Candida species, especially Candida albicans. In this study, the effect of carotenoids in carrot extracts on Candida species including Candida albicans ATCC1677, Candida glabrata CBS2175, Candida parapsilosis ATCC2195, and Candida tropicalis CBS94 was investigated. METHODS: In this descriptive study, the carrot plant was prepared from a carrot planting site in December 2012, and then the characteristics of the plant were determined. After extracting carotenoids from carrots, the susceptibility of different Candida species to carotenoids in carrot extract was determined. Also, the minimum inhibitory concentration and the minimum lethal concentration of the extracts were measured by the macro-dilution method. Finally, the data were analyzed by SPSS software using the Kruskal-Wallis test and Mann-Whitney post-hoc test with Bonferroni adjustment. RESULTS: The highest growth inhibition zone was obtained for carrot extract at a concentration of 500 mg/ml for C. glabrata and C. tropicalis. The MFC of carrot extract on Candida species was 62.5 mg/ml for C. albicans, C. glabrata, and C. parapsilosis, and 125 mg/ml for C. tropicalis. The MFC of carrot extract on Candida species was 125 mg/ml for C. albicans, C. glabrata, and C. parapsilosis, and 250 mg/ml for C. tropicalis. CONCLUSION: The present study can be the starting point for research activities in this direction and promises new therapies based on the use of carotenoids.

Immunomodulatory drug fingolimod (FTY720) restricts the growth of opportunistic yeast Candida albicans in vitro and in a mouse candidiasis model.

Fingolimod (FTY720) is a drug derived from the fungicidal compound myriocin. As it was unclear whether FTY720 has antifungal effects as well, we aimed to characterize its effect on Candida albicans in vitro and in a mouse candidiasis model. First, antifungal susceptibility testing was performed in vitro. Then, a randomized, six-arm, parallel, open-label trial was conducted on 48 mice receiving oral FTY720 (0.3 mg/kg/day), intraperitoneal C. albicans inoculation, or placebo with different combinations and chorological patterns. The outcome measures of the trial included serum concentrations of interleukin-10 and interferon-gamma, absolute lymphocyte counts, and fungal burden values in the mice's livers, kidneys, and vaginas. Broth microdilution assay revealed FTY720's minimum inhibitory concentration (MIC99) to be 0.25 mg/mL for C. albicans. The infected mice treated with FTY720 showed lower fungal burden values than the ones not treated with FTY720 (p<0.05). As expected, the mice treated with FTY720 showed a less-inflammatory immune profile compared to the ones not treated with FTY720. We hypothesize that FTY720 synergizes the host's innate immune functions by inducing the production of reactive oxygen species. Further studies are warranted to unveil the mechanistic explanations of our observations and clarify further aspects of repurposing FTY720 for clinical antifungal usage.

Comparative assessment of the mouse immune responses to colistin-resistant and colistin-sensitive isolates of Acinetobacter baumannii.

Acinetobacter baumannii is known as the most frequent species in clinical samples that is responsible for a large number of nosocomial infection outbreaks. The colistin-resistance of this bacterium has been found to be increasing. This study aimed to evaluate the immune response to colistin-susceptible and colistin-resistant A. baumannii isolates in a mouse model. Samples were prepared from the wounds of patients suspected of A. baumannii admitted to the intensive care unit of Namazi Hospital in Shiraz. Antibiotic susceptibility was studied by disk diffusion and broth microdilution according to CLSI and EUCAST criteria. Among the isolates, one colistin-sensitive isolate and one colistin-resistant isolate were injected intraperitoneally to BALB/C mice. Blood samples were collected after 4 h and cytokines (IL1-ß, IL-12, IFN γ, and IL-10) and surface markers (CD4, CD8, CD3, and CD45) were determined by ELISA and flow cytometry. Then, hematologic and histopathological factors were analyzed, and colony count in lung, liver, kidney, and spleen tissues were also performed. The results showed that levels of cytokines (IL1-ß, IL-12, IFN γ, and IL-10) and markers (CD4, CD8, CD3 and CD45) were higher in mice receiving colistin-resistant A. baumannii isolates than in mice receiving colistin-sensitive A. baumannii isolates, indicating that colistin-resistant isolates 4 h following the intraperitoneal injection stimulated host innate immune system better and produced a stronger immune response. On the other hand, histopathological findings showed inflammatory cell infiltration, hyperemia, and tissue damage. In addition, the bacterial load and tissue damage in the lung was higher than other tissues. The results of this study can have promising potential for the development of a prevention and treatment strategy based on cellular immune response for infection caused by colistin-sensitive and colistin-resistant A. baumannii.

Molecular investigation of the incidence of Candida auris infections at selected hospitals in Iran.

BACKGROUND: The accurate occurrence rate of C. auris infections is still not clear, mainly due to the defects in detection and identification tools routinely used. In this study, we used conventional PCR and real-time PCR assays for sensitive and specific detection/identification of C. auris from either yeast isolates or clinical specimens collected from various patients in different parts of Iran. Our survey is the first large-scale study rating the incidence of C. auris infections in Iran. METHODS: A total of 439 yeast isolates and 590 clinical specimens were screened by specific C. auris-PCR, targeting the ITS region. The validity of positive samples was assessed by sequencing. RESULTS: Four out of 590 clinical specimens (0.68%) were positive by conventional PCR, while in real-time PCR performed on 100 clinical samples, including those four samples positive in conventional samples, 6 samples were positive. A complete agreement of the identification of positive cases with sequencing results was documented. Among 439 culture isolates, none was positive for C. auris. After following up and resampling of the patients with positive PCR, only one specimen showed positive culture for C. auris, which was confirmed by sequencing. CONCLUSION: C. auris is not a common cause of systemic or superficial fungal infections in Iran, and a few detected positive cases can be considered as a commensal, coloniser or infecting yeast which may potentially emerge in some clinical and therapeutical conditions. Mycological and phenotypical assays are not sensitive approaches for isolation/identification of C. auris, unless a specific and sensitive molecular-based method is applied.

WITHDRAWN: Evaluating the Antifungal Activity of Rumex acetosella, Teucrium polium, and Glycyrrihize globra var. violacca on Pathogenic Dermatophytes and Determining Phenolic Compounds

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A Chronic Autochthonous Fifth Clade Case of Candida auris Otomycosis in Iran.

Candida auris, a multidrug-resistant nosocomial pathogen, has emerged globally with high morbidity and mortality among immunocompromised individuals and COVID19 hospitalized patients. Five major clades of C. auris have been previously described. The fifth clade is exclusively found in Iran where C. auris isolates are genetically distinct from other clades by > 200,000 single-nucleotide polymorphisms. The origin of C. auris remains unclear, and limited clinical data are available at present regarding clade V infection or colonization. Herein, another case of otomycosis in Iran caused by an isolate of C. auris belonging to the fifth clade is reported. Genotyping revealed that the obtained C. auris isolate from Isfahan clustered with earlier clade V isolates from Babol, cities around 600 km separated, which indicates that C. auris clade V is established in Iran. C. auris is thought to exist more commonly in Iran, given that limited diagnostic capacity in the country has probably curbed the identification of more C. auris cases. Therefore, surveillance of the environment, patients and healthcare facilities in different geographical regions in Iran is urgently required.

Evaluation antibiotic resistance and presence of blaOXA-51, blaOXA-58 and blaOXA-23 genes in Acinetobacter baumannii strains via multiplex PCR.

The resistance of Acinetobacter baumannii to most antibiotics is increasing. The presence of metallo-beta-lactamase and carbapenemase enzymes has led to the resistance of these bacteria to carbapenems as one of the major classes of broad-spectrum antibiotics and has raised concerns in human societies. This research evaluated the presence of blaOXA-51, blaOXA-58 and blaOXA-23 genes in A. baumannii strains during a 12 months period. One hundred strains were isolated from the patients hospitalized in ICU of Ali Asghar and Shahid Rajaee trauma hospitals in Shiraz. Bacterial identity was determined by biochemical tests and antibiotic resistance was determined by disk diffusion method. The isolated strains were then evaluated in terms of carrying blaOXA-23, blaOXA-51 and blaOXA-58 genes, using the multiplex PCR method. The results showed that A. baumannii was resistant to carbapenems but most strains were susceptible to tigecyclin and colistin. The majority of strains carried the blaOXA-23 and blaOXA-51 genes, but very few carried the bla OXA-58 gene. The results revealed that the antibiotic resistance of A. baumannii is increasing, which causes a more outbreak of this organism.

Detection of fungal and bacterial contamination of hazelnut and determination of aflatoxin B by HPLC method in Isfahan, Iran.

Background and Purpose: Due to the fact that fungal species, such as Aspergillus flavus and Aspergillus parasiticus produce carcinogenic and mutagenic aflatoxins and have the potential to produce fungal secondary metabolites, fungal contamination should be avoided. This study was conducted using the HPLC method and aimed to examine the fungal contamination of Isfahan hazelnuts in order to identify the presence of Aflatoxins. Materials and Methods: In total, 100 samples of hazelnuts were randomly collected from supermarkets in Isfahan. The samples were then cultured on Sabouraud dextrose agar media and analyzed to determine fungal contaminations. The aflatoxin analysis was carried out using the HPLC method. Results: It was discovered that nine genera of fungi, namely Aspergillus, Penicillium, Rhizopus, Ulocladium, Alternaria, Drechselera, Trichothecium, Scopulariopsis, and Mucor were identified in 78% of the samples. Samples contaminated with Aspergillus flavus (22 samples) were studied to determine the presence of aflatoxin. The results showed that 16 (72.72%) of the samples were contaminated with AFB1, AFB2, and AFG2 and the mean concentrations were 0.926, 0.563, and 0.155 ng/g, respectively. Conclusion: Some parameters that affect mycotoxin production are temperature, food substrate, the strain of the mold, and other environmental factors. Due to the toxigenic quality of some of these fungi and their hazard to human health, it is crucial that fungal contamination and aflatoxin identification tests are carried out before certain products are made available to the mass market.

Sonication alkaline-assisted preparation of Rhizopus oryzae biomass for facile bio-elimination of tetracycline antibiotic from an aqueous matrix.

The present study aimed to remove tetracycline (TET) antibiotic molecule from an aqueous medium using adsorbents prepared from Rhizopus oryzae biomass. The TET adsorption process was discontinuous and the adsorbent biomass was crude and NaOH-sonication-modified Rhizopus oryzae fungi. Specific active surface area for crude and modified Rhizopus oryzae was 10.38 m2/g and 20.32 m2/g, respectively. The results showed that the maximum TET adsorption efficiency was determined at pH 4, temperature 25 °C, initial TET concentration 10 mg/L, contact time 80 min, and biomass quantity 2 g/L. The equilibrium behavior showed that the Langmuir model suitably described the process. The maximum TET adsorption capacity was determined to be 38.02 mg/g and 67.93 mg/g, respectively, indicating that the method of biomass modification promoted the bio-adsorption capacity. A higher correlation coefficient (R2) and lower RMSE for the pseudo-first-order kinetic than other models showed its ability to describe the behavior of TET bio-adsorption. The enthalpy thermodynamic parameter (ΔH°) for the TET adsorption process was determined - 63.847 kJ/mol and - 85.226 kJ/mol for the raw and modified Rhizopus oryzae, respectively. Therefore, it can be suggested that the biomass of Rhizopus oryzae especially the modified version can be effectively used for the TET removal from aqueous environments.

Two-step optimization process for grass hydrolysate application as biodiesel feedstock with novel quality characteristics.

A major obstacle to biodiesel commercialization is supplying feedback which increases production costs. The potential of some oleaginous yeast for conversion of waste materials to biodiesel feedstock can overcome this problem. In this study, a potential oleaginous yeast strain was used for single-cell oil (SCO) production. Two sets of experiments were designed for the optimization process. According to the results obtained from the first experiment, lipid production and lipid content of this strain increased from 1.96 g/L and 22.6% to 3.85 g/L and 35.18% by optimization of grass hydrolysis, respectively. The results of the second experiment indicate an increase in SCO production and lipid content to 7.28 g/L and 56.39%, respectively. These results were obtained when HNO3 was used for substrate pre-treatment. Lipid analysis by gas chromatography-mass spectrometry showed a suitable and high potential of fatty acid profile for biodiesel production, which was then confirmed by evaluating the physicochemical properties of the biodiesel obtained in compliance with the US and EU standards. Consumption of microbial oil and low-cost substrate can compensate the high costs of feedstock in biodiesel production.

Aberrant expression of miR-9 in benign and malignant breast tumors.

PURPOSE: MicroRNAs (miRNAs) are involved in the progression of breast cancer (BC). miR-9 has been reported to be correlated with either favorable or unfavorable events in BC. This study was aimed to evaluate the expression level of miR-9 in human breast tissues, including benign and malignant tumor samples and also healthy tissue. MATERIALS AND METHODS: The expression level of miR-9 was analyzed in 10 normal breast tissues, 30 malignant, and 30 benign breast tumor tissue samples using RT-PCR and qPCR. In addition, bioinformatics assessment upon miR-9 functionality in BC cells was performed. RESULTS AND DISCUSSION: The miR-9 expression level was downregulated in tumor tissues, including benign and malignant compared to the healthy tissue was observed (P value, < 0.0001; fold change, -1.37). In addition, miR-9 expression level was reduced in benign tumors compared with malignant tumors (P value, < 0.0001; fold change, -1.35). Moreover, according to the AUCs (area under curve) of receiver operating characteristic (ROC) curves, miR-9 showed significant capability for distinguishing benign from healthy, malignant from healthy, benign from malignant, and tumor from health tissues. Furthermore, pathways in cancer, p53 signaling pathway, and focal adhesion were manifested by computational analysis as miR-9 related signaling pathways which have logical association with experimental observations. CONCLUSION: In conclusion, downregulation of miR-9 in benign tumors vs healthy tissue and its overexpression in malignant tumors vs benign tumors suggest paradoxical functionality for this miRNA. Our results shed additional information on controversial expression pattern of miR-9 depending on different progression level of BC.

Characterization of Diarrheagenic Antimicrobial Resistant Escherichia coli Isolated From Pediatric Patients in Tehran, Iran.

BACKGROUND: Acute infectious diarrhea is one of the most important causes of morbidity and mortality worldwide. OBJECTIVES: The objective of this study was to characterize antimicrobial resistant diarrheagenic Escherichia coli strains isolated from diarrheic children in Tehran, IR Iran. PATIENTS AND METHODS: In total, 550 stool samples from diarrheic pediatric patients, aged less than 60 months, were collected and immediately transferred to the laboratory. Isolation and identification of E. coli strains was done using bacteriological methods. Antimicrobial susceptibility testing was performed using the disk diffusion technique. Multiplex PCR was used to detect aadA1, tetA, tetB, dfrA1, qnr, aac (3)-IV, sul1, blaSHV, CITM, cat1, and cmlA antibiotic resistance genes. RESULTS: From the total of 550 fecal samples examined, 154 samples (28%) were positive for diarraheagenic E. coli. High rates of antibiotic resistance were seen against penicillin ï´¾100%), ampicillin ï´¾89.6%ï´¿ and tetracycline ï´¾83.1%ï´¿. Resistance against ciprofloxacin was low ï´¾28.6%ï´¿. The prevalence of different resistance genes in the studied strains varied from 96.10% for aadA1 gene to 40.25% for sul1 gene. The frequencies of aadA1, tetA, tetB, dfrA1, qnr, aac(3)-IV, sul1, blaSHV, CITM, cat1, and cmlA genes were 96.10%, 85.06%, 84.41%, 51.94%, 72.07%, 54.54%, 40.25%, 57.79%, 90.25%, 59.74% and 60.38%, respectively. CONCLUSIONS: Our results indicated that antibiotic resistance is increasing in diarraheagenic E. coli strains in Iran. It is imperative to develop strategies for prevention and control of resistant organisms. Changes in patterns of resistance against commonly used antibiotics in Iran indicate that an applied surveillance system and introduction of guidelines for appropriate antibiotic prescription are necessary.

Uropathogenic Escherichia coli in Iran: serogroup distributions, virulence factors and antimicrobial resistance properties.

BACKGROUND: Urinary tract infections (UTIs) are one of the most common bacterial infections with global expansion. These infections are predominantly caused by uropathogenic Escherichia coli (UPEC). METHODS: Totally, 123 strains of Escherichia coli isolated from UTIs patients, using bacterial culture method were subjected to polymerase chain reactions for detection of various O- serogroups, some urovirulence factors, antibiotic resistance genes and resistance to 13 different antibiotics. RESULTS: According to data, the distribution of O1, O2, O6, O7 and O16 serogroups were 2.43%, besides O22, O75 and O83 serogroups were 1.62%. Furthermore, the distribution of O4, O8, O15, O21 and O25 serogroups were 5.69%, 3.25%, 21.13%, 4.06% and 26.01%, respectively. Overall, the fim virulence gene had the highest (86.17%) while the usp virulence gene had the lowest distributions of virulence genes in UPEC strains isolated from UTIs patients. The vat and sen virulence genes were not detected in any UPEC strains. Totally, aadA1 (52.84%), and qnr (46.34%) were the most prevalent antibiotic resistance genes while the distribution of cat1 (15.44%), cmlA (15.44%) and dfrA1 (21.95%) were the least. Resistance to penicillin (100%) and tetracycline (73.98%) had the highest while resistance to nitrofurantoin (5.69%) and trimethoprim (16.26%) had the lowest frequencies. CONCLUSIONS: This study indicated that the UPEC strains which harbored the high numbers of virulence and antibiotic resistance genes had the high ability to cause diseases that are resistant to most antibiotics. In the current situation, it seems that the administration of penicillin and tetracycline for the treatment of UTIs is vain.