Bipolaris Spicifera Research Articles

First report of Curvularia spicifera (≡ Bipolaris spicifera) causing spathe blight and leaf spot disease of pearl millet (Cenchrus americanus) in India.

Cenchrus americanus (L.) Morrone (Poaceae), is an important millet crop cultivated mainly in arid and semiarid regions and is a staple food grain for millions of people. During 2021 July surveys in the pearl millet fields in Mysore (12°30'55" N; 76°56'54" E), Karnataka, India, plants showed spathe blight and leaf spot disease with an overall incidence ranging from 5 - 8% in the 15 hectares surveyed. Infected leaves appeared brown, and lesions extended to the sheath. Some spathes were also found infected with similar symptoms. Diseased leaves and spathes were collected (n = 5 each) for pathogen identification. Samples were cut into small pieces (0.5 cm2), sterilized with sodium hypochlorite (2%, v/v), and blotted dried. The associated fungal pathogen was isolated on potato dextrose agar (PDA) medium amended with Streptomycin (40 mg/L) and incubated at 28 ºC for 1 week. Colonies were grey, fluffy, cottony with an irregular margin, undulate and dark brown in the back of the plate. Conidiophores were pale brown, erect, slightly curved, septate, unbranched, verruculose and measured 27.1 - 94 µm in length × 2.3 - 4.5 µm in width (n = 20). Conidiogenous cells were brown, subcylindrical, irregularly shaped, and conidia were straight, mainly elliptical, dark brown smooth, with two to three septa, with measurements of 11.1 - 26.4 µm by 5.7 - 14.3 µm (n = 50). Based on morphological characters, the pathogen was identified as Curvularia sp. Two representative isolates (UOMPM1 & UOMPM2) were molecularly identified. The total genomic DNA was extracted with a CTAB method, and ITS, GAPDH and tef-1α loci were amplified using primers ITS1/ITS4 (White et al., 1990), GPD1/GPD2 (Berbee et al., 1999) and EF1983F/EF-2218R (Schoch et al., 2009) respectively. ITS sequence had 100% similarity (706/706bp) with reference sequence C. spicifera (MH863648; HF934915 & HF934916); tef-1α sequence had 100% (933/933bp) identity with C. spicifera (KM062878, KJ939505), and the GAPDH sequence was 99.8% identical to that of Curvularia sp. (MG979055), and C. spicifera (MH809681). Combined dataset of concatenated sequence (ITS-GAPDH-tef-1α) was used in a phylogenetic analysis and revealed that the isolates were in a common clade with the isolate of Curvularia spicifera (CBS 274.52) thus, confirming the identity of the isolated pathogen as C. spicifera. The sequences obtained in the present study were deposited in the GenBank (ITS: OQ253406, OQ253407; LSU: OQ253429, OQ253430; GAPDH: OQ263372, OQ263373 & TEF: OQ263374, OQ263375). Pathogenicity test was carried out by inoculating (foliar /whole plant spray) 60 healthy pearl millet plants (45-days old), grown in field plot with spore suspension (105 conidia/ml). Control plants (n=20) were treated with sterile water. The experiments were conducted in triplicates and repeated twice. Development of disease symptoms was recorded on 41 plants, and all control plants remained healthy. The identity was confirmed after re-isolation as C. spicifera based on cultural and molecular sequence analysis. To our knowledge, this is the first report of C. spicifera causing a leaf spot and spathe blight disease of pearl millet in India. This disease seriously affects grain production, and effective disease management strategies need to be investigated.

Secondary metabolites with fungicide potentials from the deep-sea seamount-derived fungus Talaromyces scorteus AS-242

Marine natural products play an important role in biopesticides. Seven new secondary metabolites with different structural classes, including two cycloheptapeptides, scortide A (1) and scortide B (2), two 19-nor-diterpenoids, talascortene H (3) and talascortene I (4), two diterpenoid acids, talascortene J (5) and talascortene K (6), and one triterpenoid, talascortene L (7) were isolated and identified from the sea-anemone-derived endozoic fungus Talaromyces scorteus AS-242. Their structures were comprehensively assigned by spectroscopic data analysis, single-crystal X-ray diffraction, tandem mass spectrometry, and electronic circular dichroism (ECD) calculations. The result of the antimicrobial assay demonstrated that compounds 1 − 6 have inhibitory activity against several human, aquatic, and plant pathogens with minimum inhibitory concentration (MIC) values ranging from 1 to 64 μg/mL. Specially, compounds 2 and 4 showed significant activities against the pathogenic fungus Curvularia spicifera with the MIC value of 1 μg/mL, providing an experimental basis of 2 and 4 with the potential as lead compounds to be developed into biopesticides.

Biofilm containing the Thymus serpyllum essential oil for rice and cherry tomato conservation.

Fungal pathogens cause major yield losses in agriculture and reduce food quality and production worldwide. To evaluate new safer alternatives to chemicals for disease management and preserve the shelf life of food, this research was conducted to: determine the chemical composition of the essential oils (EOs) of Thymus serpyllum and Thymus piperella chemotypes 1 and 2; investigate the antifungal potential of EOs in vitro against: Alternaria alternata, Bipolaris spicifera, Curvularia hawaiiensis, Fusarium oxysporum f. sp. lycopersici, Penicillium italicum, Botryotinia fuckeliana; evaluate a natural T. serpyllum extract biofilm to conserve rice grain and cherry tomatoes. EOs were analyzed by GC-MS+GC-FID. EOs' antifungal activity was evaluated by dissolving Thymus extracts in PDA. Petri dishes were inoculated with disks of each fungus and incubated at 25°C for 7 days. The T. serpyllum EO displayed the best Mycelial Growth Inhibition. The antifungal effect of the T. serpyllum EO biofilm was evaluated on rice caryopsis. Disinfected grains were dipped in a conidial suspension of each fungus and sprayed with EO (300 and 600 μg/mL) prepared in Tween 20. Grains were stored. The percentage of infected grains was recorded for 30 days. The T. serpyllum EO effect on cherry tomato conservation was evaluated in vivo. Wounded fruit were immersed in the T. serpyllum EO (300 and 400 μg/mL) and inoculated with Fusarium oxysporum f. sp. lycopersici. Fruit were evaluated for 7 and 14 days. Chemical profiles thymol/carvacrol for T. serpyllum, carvacrol for T. piperella Tp1 and thymol for T. piperella Tp2 were defined. The three evaluated EOs reduced all the studied phytopathogens' fungal growth. The T. serpyllum biofilm was effective with rice storage and against Fusarium oxysporum f. sp. lycopersici for extending the shelf life of tomatoes in warehouses and storing postharvest cherry tomatoes. We suggest applying these EOs as biofilms for safe food conservation to replace synthetic products.

Antifungal efficiency of amla leaves against Alternaria alternata and Bipolaris spicifera

In the present study, have been investigated for antifungal potential against two phytopathogens namely and . Inhibitory potential of methanolic extract of the plant was analyzed by poisoned food technique in which different concentrations of test material were prepared in sterilized potato dextrose agar. Results of the investigation revealed that has potential activity against both the test pathogens with IC value of 1±0.02 mg/mL and 0.95±0.019 mg/mL against and respectively. To conclude, leaves of amla possess potential antifungal activity and may be used as a resource of biocides in pesticide and food industry.

The Effect of Fungi Accompanying cuscuta on the Germination Rates of Ocimum and cuscuta Seeds and Their Molecular Diagnosis

This study was conducted at the Faculty of Agricultural Engineering Sciences / Plant Protection Department / University of Baghdad-Jadriya, with the aim of isolating fungal pathogens from samples of the parasite on different plant families (Solanum, Astragalus, Conocarpus, chenopodium, Alhagi, Ocimum) from several areas in Baghdad and Diyala Governorate and diagnosing them phenotypically and testing their pathogenicity. The results of isolation and phenotypic diagnostics from the stems and seeds of the cuscuta showed the presence of 12 genera belonging to nine species : Alternaria, sp. (F2), Alternaria sp. (F4), Aspergillus sp. (F6), Aspergillus sp. (F12), Chaetomium sp. (Amesia sp.) (F7), Cladosporium sp. (F8), Curvularia sp. (F5), Helminthosporium sp. (F10), Penicillium sp. (F9), Rhizoctonia solani (F1), Stemphylium sp. (F3), Stemphylium sp. (F11). The results showed that the fungus Aspergillus sp., Curvularia sp., Chaetomium sp. Helminthosporium sp. and SP. Stemphylium was first recorded on the Cuscuta plant in Iraq. The results of the pathogenetic assessment showed that all the tested isolates (F1-F12) were pathogenic to Porter and Ocimum seeds, as they caused a significant reduction in seed germination rates compared to the control treatment (without inoculation with fungal vaccine). it was found that there was a difference between fungal isolates in reducing the percentage of seed germination, as some fungal isolates achieved the highest reduction in the percentage of germination of carrier seeds. the effect of their fungal filters on the germination of Porter and Ocimum seeds was tested and diagnosed in part. The results showed that the leached fungus Chaetomium sp. With concentrations of 20, 40 and 60%, it significantly reduced the percentage of germination of Porter seeds and differed significantly with the coefficients of Aspergillus SP.(F6) fungal filters, Stemphylium sp.(F3), Alternaria sp.(F4), Curvularia sp.(F5) by analogy with the comparison treatment, while there are no significant differences between the fungal filtrate at the three concentrations. The results also showed that the fungus leached Chaetomium sp.(F7). And Aspergillus sp. sp.(F6) in concentration, it had a significant effect in reducing the percentage of germination of Ocimum seeds compared to the comparative treatment. She showed that the isolates belong to fungi (F2 Aspergillus Flavus and(F4) Alternaria alstroemeria and that F5 Curvularia spicifera and F3 Stemphylium vesicarium and F7 belong to Amesia atrobrunneum and the name was changed instead of the old name Chaetomium atrobrunneum and deposited in the gene bank Gene Bank and NCBI under the accession numbers oq746923, Oq746924, Oq746925 and Oq74692.

Whole-genome sequences of Bipolaris bicolor, Curvularia hawaiiensis, Curvularia spicifera, and Exserohilum rostratum isolated from rice in Burkina Faso, France, Mali, and Pakistan.

Different fungal species of the Pleosporaceae family infect rice, causing similar symptoms. Reference genomic sequences are useful tools to study the evolution of these species and to develop accurate molecular diagnostic tools. Here, we report the complete genome sequences of Bipolaris bicolor, Curvularia hawaiiensis, Curvularia spicifera, and Exserohilum rostratum.

Fungal microbiota isolated from native stingless bee species inhibited pathogens of Apis mellifera

Social bees can establish interactions with microorganisms to keep their colonies free of pathogens and parasites by developing different protection strategies. We explored the fungal microbiota isolated from three species of stingless bees, Tetragonisca fiebrigi, Plebeias sp., and Scaptotrigona jujuyensis, and its potential ability to suppress pathogenic microorganisms of A. mellifera, namely Paenibacillus larvae, Ascosphaera apis and Aspergillus flavus, which were tested and evaluated. Six filamentous fungal strains, Trametes hirsuta, Alternaria alternata, Curvularia spicifera, Skeletocutis sp., Alternaria tenuissima, Monascus spp., as well as the yeast Wickerhamomyces anomalus, were selected for trials and isolated from the heads of foraging bees. The fungal strains were identified by macroscopic and microscopic taxonomic characteristics and by sequencing of the ITS1–5.8S–ITS2 region of ribosomal DNA. All fungal strains inhibited these pathogens of A. mellifera. We also evaluated the effect of the secondary metabolites extracted with and without ethanol. Both metabolites showed antimicrobial properties, and our results suggest that fungi isolated from stingless bees produce bioactive compounds with antibacterial and antifungal effects that could be used to treat Apis mellifera colony diseases and maintain colony health.

Detection of Extracellular Enzymes Produced by Fungi Isolated from Dried Fruits

Screening of 57 fungal isolates isolated from dried fruits for the production of lipase, cellulase, invertase, and protease showed that there is a variation in enzyme production not only among the different genera and species, but also among the different isolates in the same species. Thirty two of tested fungi (66%) belonging to A. flavus, A. fumigatus, A. niger, A. terreus, A. tamarii, Cochliobolus spicifer, Humicola insolens, F. oxysporium, P. glaprum, P. oxalicum, P. stekii, P. variabile and Phoma sp. had a high ability to produce lipase. Ten of tested isolates representing 24.39% of tested fungi had a moderate ability to produce cellulase enzyme. These isolates were belonging to Aspergillus flavus, A. niger, A. parasiticus, A. versicolor, Curvularia 
 lunata, Penicillium griseofulvum, P. oxalicum, P. stekii, P. variabile and Ulocladium atrum. Twenty seven isolates representing 37.64 % of 
 tested isolates belonging to A. flavus, A. fumigatus, A. niger, A. parasiticus, A. terreus, A. versicolor, Fusarium oxysporium, Mucor 
 fuscus, P. glaprum, P. griseofulvum, P. oxalicum, P. variabile, Phoma sp., and U. atrum were moderate invertase producers. Sixteen fungal 
 isolates representing 28.07 % of tested isolates were moderate protease producers, these isolates were belonging to A. flavus, A. 
 niger, A. terreus, A. tamarii, A. versicolor, C. lunata, F. oxysporium, H. insolens, P. corylophilum, P. griseofulvum, P. oxalicum, P. stekii, 
 P. vinaceum, Rhizopus stolonifer. 

The Antifungal Properties of Tamarix aphylla Extract against Some Plant Pathogenic Fungi.

Tamarix aphylla is a Saudi herb, which possesses antimicrobial properties and potentially introduces a solution to the subsequent dilemma caused by agrochemicals and antifungal misuse. The current study aimed to assess the fungicidal properties of water and ethanolic extracts of T. aphylla leaves against Macrophomina phaseolina, Curvularia spicifera, and Fusarium spp. The chemical composition of T. aphylla was evaluated by gas chromatography/mass spectrometry technique (GC−MS) and Fourier-transform infrared spectroscopy (FTIR). The antifungal assay assessed the fungal growth inhibition using the poisoned food technique. Scanning and transmission electron microscopy (SEM and TEM) were used to evaluate the structural changes induced in the fungal species post-treatment by T. aphylla. FTIR and GC−MS analysis revealed that T. aphylla extracts were rich in aromatic and volatile compounds, such as Benzeneselenol, Gibberellic acid, and Triaziquone, which proved multiple antifungal properties. The results showed significant inhibition in the growth of all species (p < 0.05) except for F. moniliforme, where the water extract induced the highest mycelial growth inhibition at the dose of 30%. The highest inhibition was for M. phaseolina treated with the water extract (36.25 ± 1.06 mm, p < 0.001) and C. spicifera, treated with the ethanolic extract (27.25 ± 1.77 mm, p < 0.05), as compared to the untreated control and the positive control of Ridomol. SEM and TEM revealed some ultrastructural changes within the fungal growth of treated M. phaseolina, which included the thickening and mild rupture of mycelia. Those findings suggested the robust antifungal properties of T. aphylla against some filamentous fungi. The phenolic composition illustrated the potential fungicidal properties of T. aphylla. Additional studies are required to focus on more antimicrobial properties of T. aphylla against other species, particularly those that might benefit the medical field.

A Serbian endemic plant „Nepeta rtanjensis“ Diklić et Milojević – chemical composition and biological activity

Nepeta rtanjensis Diklić et Milojević as an endemic and critically endangered plant in Serbia is the object of great scientific interest. N. rtanjensis is a rare plant of a limited distribution its application in traditional medicine is practically unknown. However, bioactive potential of 4aα,7α,7aβ-nepetalactone, the main compound from volatile fraction, as well as rosmarinic and 3-O-caffeoylquinic acid from non-volatile fraction is great. Scientists found that this plant possesses good antimicrobial activities against Aspergillus sp., Bacillus cereus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Listeria monocytogenes, Micrococcus flavus, Penicillium sp., Pseudomonas aeruginosa, Salmonella sp. and Staphylococcus aureus. N. rtanjensis can be used preserving foods and beverages as well as for prevention and treatment of diseases caused by common and emerging food borne pathogens. Furthermore, antioxidant, anticancer (cervix, breast and colon carcinoma, lung adenocarcinoma and myelogenous leukemia cells) and immunomodulating activity are also noted. However, toxicity in zebrafish model was not observed, so N. rtanjensis can be classified as safe for human health. Furthermore, phytotoxic potential against Ambrosia artemisiifolia as well as activity against plant pathogens (Alternaria sp., Cladosporium cladosporoides, Bipolaris spicifera and Trichoderma viridae) are also noted. There is potential fordeveloping novel biopesticide formulations.

Phylogeny and mycotoxin profiles of pathogenic Alternaria and Curvularia species isolated from date palm in southern Tunisia.

Date palm (Phoenix dactylifera L.), is a widely cultivated crop across North Africa, with about 300 thousand tons of fruits produced per year, in Tunisia. A wide range of fungal pathogens has been associated with leaf spots of date palm, Alternaria species being the most frequently reported. Symptomatic leaves of Deglet Nour variety were randomly collected in six localities in Tunisia. We used a polyphasic approach to identify 45 Alternaria and five Curvularia strains isolated from date palm, confirming their pathogenicity. Sequencing of allergen Alt-a1, glyceraldehyde-3-phosphate dehydrogenase (gpd) and calmodulin genes allowed us to group 35 strains in Alternaria Section, and 10 strains in Ulocladioides section. Based on sequencing analyses of Internal Transcribed Spacer, gpd and elongation factor genomic regions, all Curvularia strains were identified as Curvularia spicifera. All Alternaria and Curvularia species tested on date palm plantlets proved to be pathogenic, fulfilling Koch's postulates. Although no significant differences were observed among the species, the highest mean disease severity index was observed in A. arborescens, while the lowest corresponded to C. spicifera. The capability of these strains to produce mycotoxins in vitro was evaluated. None of the A. consortialis strains produced any known Alternaria mycotoxin, whereas more than 80% of the strains included in Alternaria section Alternaria produced variable amounts of multiple mycotoxins such as alternariol, alternariol monomethyl ether, altenuene, tenuazonic acid and tentoxin. Curvularia spicifera strains produced detectable traces of fumonisins B. This work reports a first comprehensive multidisciplinary study of mycotoxigenic Alternaria species and C. spicifera associated with leaf spot disease on date palm.

Biodegradation of Petroleum Hydrocarbons by Drechsleraspicifera Isolated from Contaminated Soil in Riyadh, Saudi Arabia.

Currently, the bioremediation of petroleum hydrocarbons employs microbial biosurfactants because of their public acceptability, biological safety, and low cost. These organisms can degrade or detoxify organic-contaminated areas, such as marine ecosystems. The current study aimed to test the oil-biodegradation ability of the fungus Drechslera spicifera, which was isolated from contaminated soil samples in Riyadh, Saudi Arabia. We used hydrocarbon tolerance, scanning electron microscopy, DCPIP, drop-collapse, emulsification activity, recovery of biosurfactants, and germination assays to assess the biodegradation characteristics of the D. spicifera against kerosene, crude, diesel, used, and mixed oils. The results of DCPIP show that the highest oxidation (0.736 a.u.) was induced by crude oil on the 15th day. In contrast, kerosene and used oil had the highest measurements in emulsification activity and drop-collapse assays, respectively. Meanwhile, crude and used oils produced the highest amounts of biosurfactants through acid precipitation and solvent extraction assays. Furthermore, the biosurfactants stimulated the germination of tomato seeds by more than 50% compared to the control. These findings highlight the biodegradation ability of D. spicifera, which has been proven in the use of petroleum oils as the sole source of carbon. That might encourage further research to demonstrate its application in the cleaning of large, contaminated areas.

P332 Evaluation of the efficacy of fumigation practices on the mycological flora in the Orthopedic Operation theatre environment of a tertiary care hospital

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM ObjectivesTo determine the pre and post-fumigation prevalence of fungi in the Orthopedic Operation Theatre of a tertiary care hospital and characterization of fungal isolates.MethodThis is a prospective environmental, analytical study conducted from July 2021 to January 2022. Pre- and post-fumigation samples were taken from Ultra and Modular OT of Orthopedic department every week by passive methods of air sampling using Gravity sedimentation method (1/1/1 scheme) on SDA and also by surface sampling using swabs. The fungal load of air was measured by calculating the number of CFU per cubic meter (CFU/m3) of air by Omeliansky formula. Fumigation of OT was done with a complex formulation of stabilized hydrogen peroxide 11% w/v with silver nitrate solution 0.01% (Baccishield®).All surface samples were inoculated on SDA with chloramphenicol and all plates were incubated at 22°C and 37°C. The isolates were identified by using standard mycological procedures. Statistical analysis was done using a T-test.ResultsOut of 19 surface sampling, fumigation was found to be 100% effective only on 3 occasions (15.79%) in Ultra OT and on 7 occasions (36.84%) in Modular OT. In air sampling ≥50% reduction was found in only 4 samplings (21.05%) in Ultra OT and 10 samplings (52.63%) in Modular OT. The counts were much more than the WHO OT permissible limits. A total of 16 species of fungi were isolated belonging to 11 genera.The most common isolate was Aspergillus niger, followed by sterile hyalohyphomycetes, A. flavus, Cladosporium spp., Curvularia spp., Bipolaris spicifera, A. fumigatus, etc. in both Ultra and Modular OT.Additionally, Exophiala spp. and Fonsecaea spp. were isolated in Ultra OT.The concentrations of fungi in Ultra OT before and after fumigation were in the range 22.11-58.97 CFU/m3 and 7.37-51.59 CFU/m3, respectively. Whereas, in Modular OT, the range was 14.74-36.86 CFU/m3 and 7.37-29.48 CFU/m3, respectively. Percentage reduction of fungi following fumigation with Baccishield varied from 0% to 75% both in Ultra OT and Modular OT. The statistically correlated P-value from pre- and post-fungal concentrations in Ultra and Modular OT were found to be .002 and .0002 respectively which was found significant by T-test, albeit ineffective as per standards.ConclusionIn corroboration with our findings, Baccishield has been reported to be less effective even by other researchers. Hence, this needs to be replaced by more effective fumigants. The ineffectiveness of fumigation in Ultra OT was most probably due to the lack of HEPA filters and not strictly following up of aseptic protocols. In Modular OT improper maintenance of ACs and lack of periodic cleaning up of HEPA filters may be contributing factors.Hence, implementing more stringent, frequent, and comprehensive disinfection and cleaning procedures, educating and motivating the health care personnel can help to improve the air quality of OTs that may aid in reducing post-operative infections.

Biocontrol of okra-rot-causing Cochliobolus spicifer-CSN-20 using secondary metabolites of endophytic fungi associated with Solenostemma arghel

Rot disease is responsible for serious economic losses related to okra (Abelmoschus esculentus) crops cultivated in Upper Egypt. Colonies with a consistent morphology were isolated from the infected okra stems and leaves and subjected to morphological and molecular examinations. The causal pathogen was identified as Cochliobolus spicifer based on morphological fungus descriptions, as well as on the amplified 28S rDNA and internal transcribed spacer region (ITS) sequences, which showed 99%–100% similarity to the sequences of C. spicifer-CSN-20 strains. The volatile and non-volatile organic compounds (VOCs and n-VOCs, respectively) produced by the endophytic fungi that are associated with the medicinal plant Solenostemma arghel, namely Fusarium solani-F4-1007, Penicillium verrucosum-F2-1006, and Aspergillus terreus-F5-1008, inhibited the growth of the C. spicifer pathogen by 34.2%,31.4%, and 30.5%, respectively. In total,27 VOCs were identified by GC/MS, among which eight were specific to A. terreus-F5-1008, eight to P. verrucosum-F2-1006, and three to F. solani-F4-1007, whereas nine VOCs were commonly produced by the three endophytic fungi. Moreover, F. solani-F4-1007-produced VOCs and n-VOCs exhibited the highest antifungal activity, with 37.27% and 37.1% inhibition against C. spicifer colony growth, respectively. The potent antifungal VOCs produced by F. solani-F4-1007 were identified as 3,4-dihydro-2 h-1,5-(3″-t-butyl) benzodioxepine, 4-(2-hydroxyethyl) phenol, and phenylethyl alcohol using GC/MS. Therefore, F. solani-F4-1007 was tested as a potential biocontrol agent against C. spicifer-CSN-20 using an in-planta assay. Okra plants treated with endophytic F. solani-F4-1007did not show any disease symptoms, whereas those that were not treated with F. solani-F4-1007 exhibited severe disease symptoms when challenged with inoculation of the C. spicifer pathogen. Our results demonstrated the contribution of the endophytic fungus F. solani-F4-1007 as a potential biocontrol agent against the C. spicifer pathogen, to improve okra growth.

Soil mycoflora of Thi-Qar marshes and their enzymatic activities

A survey on soil mycoflora in Thi-Qar marshes have been studied. A total of 22 fungal species belonging to 13 genera were isolated and identified. The average of the total fungal colony count ranged between 1550-4150 colonies/gm of soil. Aspergillus, Alternaria, Cladosporium, Stachybotrys and Phoma were of the mostly occurred genera in soil samples constituting 98.67%, 82.67%, 69.33%, 57.0% and 52.0%, respectively, and the mostly occurred species were; A. terreus, A. flavus, C. cladosporioides and S. atra; constituting 74.67%, 69.33%, 69.33% and 57%, respectively. Nine fungal species: Alternaria chlamydospora, Aspergillus flavus, A. fumigatus, A. terreus, Bipolaris spicifera, Fusarium oxysporum, Exserohilum rostratum, Trichoderma viride and Ulocladium botrytis were identified to produce the four investigated enzymes: cellulase, amylase, lipase and protease. All the tested isolates were identified as cellulase and amylase producers, A. candidus, A. flavus, B. spicifera and C. cladosporioides were an excellent producer of cellulase, while A. terreus, Stachybotrys atra and T. viride were the most active amylase producers. A. flavus, F. oxysporum and U. atrum showed the highest lipase production. The higher activity of protease was produced by A. candidus, A. flavus and T. viride.&#x0D; &#x0D;

Rare cases of systemic phaeohyphomycoses caused by Bipolaris species as a Post SARS-COV 2 sequelae

PurposeThe deadly second wave of COVID-19 brought an unprecedented rise of cases of associated invasive fungal infections. In this study we present two rare cases of systemic phaeohyphomycoses by Bipolaris species as a Post SARS-COV 2 sequelae, which may usually be neglected as contaminants.Methods & MaterialsRetrospective study of 1150 specimens received from patients with a high index of clinical suspicion of invasive fungal infections from April 2021 to June 2021 to identify cases of invasive phaehyphomycoses among post COVID patients. Diagnosis of phaeohyphomycoses was established on microbiological evidence including microscopy and culture, and radiological evidence along with supporting clinical features. The diagnosis of phaeohyphomycoses was confirmed when dark pigmented thin septate hyphae with or without spores were demonstrated in aseptically aspirated fluid or tissue specimen with pure culture isolate of velvety dark, brownish black flat colonies on Sabouraud dextrose agar at 25°C.ResultsDuring the study period, two cases were confirmed as invasive phaeohyphomycoses caused by Bipolaris spicifera phenotypically. The first case was of invasive fungal sinusitis and second of invasive pulmonary phaeohyphomycoses. Overall incidence rate was 0.55% (2/364). Both cases were post COVID, with history of COVID 2-3 weeks before current presentation and hospitalisation with oxygen support for the same, uncontrolled diabetes mellitus (HbA1C: 11 & 8 respectively), and hypothyroidism. First case was initially treated as of mucormycosis, considering her clinical presentation and high index of suspicion with several similar cases reported in our area. He was managed with Amphotericin B along with surgical debridement. Second case was suspected as case of invasive pulmonary aspergillosis and was managed with voriconazole initially and later started on itraconazole after laboratory confirmation. Due to initial false diagnosis in our cases, patients were subjected to long course of amphotericin B and voriconazole respectively, whereas it could have been managed with itraconazole which has far lesser side effects.ConclusionHigh index of suspicion is required to confirm invasive phaeohyphomycoses as dematiaceous fungi are often contaminants or commensals. Correct and timely diagnosis is necessary for adequate management of invasive phaeohyphomycoses cases.

Comparative study of antifungal activity of two preparations of green silver nanoparticles from Portulaca oleracea extract

The green silver nanoparticles (green AgNPs) exhibit an exceptional antimicrobial property against different microbes, including bacteria and fungi. The current study aimed to compare the antifungal activities of both the crude aqueous extract of Portulaca oleracea or different preparations of green AgNPs biosynthesized by mixing that aqueous extract with silver nitrate (AgNO3). Two preparations of the green AgNPs were synthesized either by mixing the aqueous extract of P. oleraceawith silver nitrate (AgNO3) (normal AgNPs) or either irradiation of the AgNPs, previously prepared, under 60Co γ-ray using chitosan (gamma-irradiated AgNPs). Characterization of different AgNPs were tested by Zeta potential analyzer, Ultraviolet (UV) Visible Spectroscopy, and Fourier-Transform Infrared (FTIR) spectrometry. Three different plant pathogenic fungi were tested, Curvularia spicifera, Macrophomina phaseolina, and Bipolaris sp. The antifungal activities were evaluated by Transmission Electron Microscope (TEM) for either the crude aqueous extract of P. oleracea at three doses (25%, 50%, and 100%) or the newly biosynthesized AgNPs, normal or gamma-irradiated. With a few exceptions, the comparative analysis revealed that the irradiated green AgNPs at all three concentrations showed a relatively stronger antifungal effect than the normal AgNPs against all the three selected fungal strains. UV–visible spectroscopy of both preparations showed surface plasmon resonance at 421 nm. TEM results showed that both AgNPs were aggregated and characterized by a unique spherical shape, however, the gamma-irradiated AgNPs were smaller than the non-irradiated AgNPs (0.007–0.026 µM vs. 0.009–0.086 µM). TEM photographs of the fungal strains treated with the two AgNPs preparations showed flaccid structures, condensed hyphae, and shrunken surface compared with control cells. The data suggested that the biosynthesized P. oleracea AgNPs have antifungal properties against C. spicifera, M. phaseolina, and Bipolaris sp. These AgNPs may be considered a fungicide to protect different plants against phytopathogenic fungi.

Isolation and molecular characterization of clinical and environmental dematiaceous fungi and relatives from Iran.

Background and Purpose: The frequency and genetic diversity of black fungi in environmental and clinical settings have not been fully studied in Iran. This study aimed to identify and evaluate intra- and inter-species DNA sequence variation and also understand the phylogenetic relationships of melanized fungi and relatives isolated from different geographical regions of Iran. Materials and Methods: In total, 111 clinical and environmental strains of dematiaceous fungi were isolated, and their internal transcribed spacer ribosomal DNA (rDNA) regions were sequenced and analyzed. Results: An inter-species nucleotide sequence diversity rate of 1 to 464 nucleotides was observed between the species. Intra-species differences were found in the strains of Alternaria alternata, Cladosporium cladosporioides, Alternaria tenuissima, Curvularia spicifera, Aureobasidium pullulans, Curvularia hawaiiensis, Neoscytalidium dimidiatum, Alternaria terricola, Alternaria chlamydospora, Didymella glomerata, and Drechslera dematioidea by 0–59, 0–22, 0–4, 0–4, 0–3, 0–2, 0–2, 0–2, 0–2, 0–1, and 0–1 nucleotide, respectively. Conclusion: The internal transcribed spacer rDNA is useful for the discrimination of several taxa of dematiaceous fungi. However, a better understanding of the taxonomy of species of Alternaria requires a larger rDNA region or a library of other gene sequences.

A case of cutaneous colonization of Bipolaris spicifera associated with radiation dermatitis.

A case of cutaneous colonization of Bipolaris spicifera associated with radiation dermatitis.

Endophytic Fungi, Drechslera spicifera, Mediated Synthesis of Biogenic Silver Nanoparticles

Abstract: Aim: In present communication, endophytic fungi Drechslera spicifera is isolated from leaves of Cardiospermum halicacabum and used as a reducing agent for silver nanoparticles (AgNPs) synthesis. Materials and Methods: Synthesized AgNPs were characterised using UV-vis spectroscopy, FTIR, scanning-electron microscopy (SEM) and particle size analysis. Total phenolic, flavonoid content and antioxidant potential of endophytic fungi (culture and aqueous extract), host plant Cardiospermum halicacabum (methanol and aqueous extracts) and silver nanoparticles (culture and aqueous extract mediated silver nanoparticles) was determined. Results and Conclusion: The endophytic fungi Drechslera spicifera forms stable AgNPs and also showed good antioxidant potential for pharmaceutical applications. Key words: Cardiospermum halicacabum, Drechslera spicifera, Endophytic fungi, Silver nanoparticle, Antioxidant.