Diversity of filamentous fungi associated with coral and sponges in coastal reefs of northeast Brazil

Fungi are known to form associations with various marine organisms and substrata such as sponges and corals, both as potential symbionts or pathogens. These microorganisms occupy an ecological niche that has recently attracted great attention due to their potential in either ecological or pharmaceutical advances. However, the interaction between marine invertebrates and fungi is still poorly understood, including how they are affected by anthropogenic actions. Here, we identified 89 fungal isolates through sequencing of the ITS rDNA region obtained from the various sponge and coral species collected at two northeast Brazilian reefs. We found 43 species of fungi from 16 genera, all belonging to phylum Ascomycota. The sponges and coral shared four genera: Aspergillus, Penicillium, Trichoderma, and Cladosporium, all commonly found in terrestrial habitats and associated with marine invertebrates. We observed some unusual species in relation to the marine environment, such as Clonostachys rosea and Neopestalotiopsis clavispora, most of them related to plants, either as saprophytic or pathogenic, suggesting that these species were transported from the surrounding terrestrial environment to the reefs. In addition, some isolates represent possible undescribed species, reinforcing the importance of studying the marine environment in relation to its ecological and biotechnological importance.     

Keratinophilic fungi from the vicinity of salt pan soils of Sambhar lake Rajasthan (India)

Forty soil samples were collected from seven sites in the vicinity of Sambhar lake Rajasthan, India and screened for the presence of keratinophilic fungi using hair baiting techniques for isolation. Seventeen isolates were recovered and identified. The cultures were identified by recognition of their macro- and micro- morphological features. Their identification was confirmed by BLAST using ITS1-5.8S-ITS2 rDNA region and sequences have been deposited in NCBI data base. A total of 34 species belonging to 29 genera were isolated. Among the dermatophytes and related keratinophilic fungi Chrysosporium indicum was predominant followed by Ctenomyces serratus, C. tropicum, Keratinophyton durus, Auxarthron conjugatum, Gymnascella dankaliensis, Gymnoascoideus petalosporus and Uncinocarpus reesii. Twenty-six species belonging to 22 genera represented other species. Our study indicates that keratinophilic fungi and species are found in the soils at the vicinity of the Sambhar Lake, and human activities can be the potential source of pathogenic fungi.     

Fungal endophytes in five medicinal plant species from Kudremukh Range, Western Ghats of India

Eighteen species of endophytic fungi were isolated from bark, stem and leaf segments of five medicinal plant species growing within the Kudremukh range in the Western Ghats of India. The dominant endophytic fungal species isolated from these plant species were Curvularia clavata, C. lunata, C. pallescens and Fusarium oxysporum. The highest species richness as well as frequency of colonization of endophytic fungi was found in the leaf segments, rather than the stem and bark segments, of the host plant species. The greatest number of endophytic fungal species were found within Callicarpa tomentosa (11 species), whereas Lobelia nicotinifolia harbored the lowest number of fungal endophytes (5 species). This study provides evidence that fungal endophytes are host and tissue specific.     

Generic and functional diversity in endophytic actinomycetes from wild Compositae plant species at South Sinai – Egypt

The diversity of culturable endophytic actinomycetes associated with wild Compositae plants is scantily explored. In this study, one hundred and thirty one endophytic actinobacteria were isolated from ten Compositae plant species collected from South Sinai in Egypt. Microscopic and chemotaxonomic investigation of the isolates indicated fourteen genera. Rare genera, such as Microtetraspora, and Intrasporangium, which have never been previously reported to be endophytic, were identified. Each plant species accommodated between three to eight genera of actinobacteria and unidentified strains were recovered from seven plant species.The generic diversity analysis of endophytic assemblages grouped the plant species into three main clusters, representing high, moderate and low endophytic diversity. The endophytes showed high functional diversity, based on forty four catabolic and plant growth promotion traits; providing some evidence that such traits could represent key criteria for successful residence of endophytes in the endosphere. Stress-tolerance traits were more predictive measure of functional diversity differences between the endophyte assemblages (Shannon's index, p = 0.01). The results indicate a potential prominent role of endophytes for their hosts and emphasize the potency of plant endosphere as a habitat for actinobacteria with promising future applications.     

Unusual causes of fungal rhinosinusitis: A study from a tertiary care centre in South India

Purpose: The frequency of mycotic infections of the nose and paranasal sinuses has been increasing over the past three decades. Apart from the common causes of fungal rhinosinusitis such as Aspergillus species and Penicillium species, there have been reports of rare and unusual fungi isolated from India and other countries. Objective: The objective of this study is to find out the prevalence of fungal infections of the nose and paranasal sinuses caused by unusual fungal isolates at a tertiary care teaching hospital in South India. Materials and Methods: Duration of the study period was from April 2009 to March 2010. Specimens were collected from the nose and paranasal sinuses of all clinically and radiologically diagnosed cases of rhinosinusitis. All the clinical specimens were processed by standard methods for fungal culture. This included initial screening by 10% potassium hydroxide, inoculation of the specimen onto Sabouraud dextrose agar and incubation at 25°C and 37°C, followed by slide culture and other special techniques wherever necessary. Histopathological examination was also performed for the specimens. Results: A total of 60 specimens were received for fungal culture from cases of rhinosinusitis during the period, out of which 45 showed no growth. There were nine cases of Aspergillus flavus, 1 each of Aspergillus fumigatus and Penicillium species. The rest four specimens grew rare fungal isolates, i.e. Acremonium sp., Scedosporium apiospermun, Cladosporium cladosporioides and Lasiodiplodia theobromae. Histopathological findings were also positive for these four cases. Conclusion: Apart from the common causes, unusual fungal pathogens were isolated from cases of rhinosinusitis during the study period, which is in accordance with similar reports from other parts of India and some other countries.     

Species diversity of culturable endophytic fungi from Brazilian mangrove forests

This study aimed to perform a comparative analysis of the diversity of endophytic fungal communities isolated from the leaves and branches of Rhizophora mangle, Avicennia schaueriana and Laguncularia racemosa trees inhabiting two mangroves in the state of São Paulo, Brazil [Cananeia and Bertioga (oil spill-affected and unaffected)] in the summer and winter. Three hundred and forty-three fungi were identified by sequencing the ITS1-5.8S-ITS2 region of rDNA. Differences were observed in the frequencies of fungi isolated from the leaves and branches of these three different plant species sampled from the Bertioga oil spill-affected and the oil-unaffected mangrove sites in the summer and winter; these differences indicate a potential impact on fungal diversity in the study area due to the oil spill. The molecular identification of the fungi showed that the fungal community associated with these mangroves is composed of at least 34 different genera, the most frequent of which were Diaporthe, Colletotrichum, Fusarium, Trichoderma and Xylaria. The Shannon and the Chao1 indices [H′_(95 %) = 4.00, H′_(97 %) = 4.22, Chao1_(95 %) = 204 and Chao1_(97 %) = 603] indicated that the mangrove fungal community possesses a vast diversity and richness of endophytic fungi. The data generated in this study revealed a large reservoir of fungal genetic diversity inhabiting these Brazilian mangrove forests and highlighted substantial differences between the fungal communities associated with distinct plant tissues, plant species, impacted sites and sampling seasons.     

Diversity of microfungi in the sandy soil of Ipanema Beach,Rio de Janeiro,Brazil

The mycobiota of the sandy soil of Ipanema Beach, Rio de Janeiro, Brazil, was investigated in 144 sand samples collected at four different sites along the sea coast, divided into three subsites. for a period of one year. A total of 4285 yeast colonies and of 6956 of colonies filamentous fungi were isolated using conventional media and techniques. Representatives of the filamentous fungi corresponding to a total of 1334 colonies were identified and assigned to 34 genera and 170 species. The genera of highest incidence and their respective numbers of species were as follows: Aspergillus, 30.4%, 32 spp.; Penicillium, 16.2%, 35 spp.; Fusarium, 12,6%, 33 spp.: Trichoderma, 6.4%, 7 spp.; Paecilomyces, 3.7%, 10 spp.: Cladosporium, 3.1 %, 8 spp. and Acremonium, 1.0%, 8 spp. Several other genera and species were detected at quite low occurrences. Non-sporulating fungi (18.3%) and Coelomycetes (Sphaeropsidales) (1.9%) were also detected. Most of the genera detected belonged to the Deuteroycotina, with fewer proportions belonging to the Ascomycotina and Zygomycotina. Moniliaceae was represented by the largest number of species and Dematiaceae was represented by the largest number of genera. In terms of seasonal distribution, the largest number of filamentous fungi was isolated during the winter and the smallest number the Summer.     

Characterization and genetic diversity of sugarcane streak mosaic virus causing mosaic in sugarcane

Sixty-three sugarcane leaf samples were collected from fifty-eight sugarcane varieties, evolved from eleven major sugarcane growing states in India, Australia, South Africa and USA. In RT-PCR, using gene specific primers for sugarcane streak mosaic virus (SCSMV)-CP, 58 of 63 sugarcane samples were found positive to the virus infection and rest of the five samples were negative. Partial CP gene sequences of 42 SCSMV isolates including an isolate from aphid colony (Melanaphis indosacchari) infested on sugarcane variety from this study were characterized after cloning and sequencing for selective isolates represented by at least one isolate from each location. The new sequences identified in the study were named as SCSMV-CB isolates. Fifty two sequences including the 10 database sequences (complete CP cds) deposited earlier from this institute were compared with each other as well as GenBank database sequences of Potyviridae members viz., Rymovirus, Potyvirus, Ipomovirus, Tritimovirus and eight sequences of SCSMV reported from elsewhere. Among the SCSMV-CB isolates sequenced in the study, 85.7–100% (nucleotide) and 89.9–100% (amino acid) sequence identities were observed and with the other data base sequences of SCSMV, the respective identities were 82.2–97.5 and 89.7–98.6%. Grouping of the isolates by the maximum likelihood with molecular clock model, distributed 60 SCSMV sequences including the eight database sequences deposited by other SCSMV working groups from India and USA in 16 different phylogenetic groups. Although the isolates of SCSMV were relatively close to Ipomovirus and Tritimovirus, they were sandwiched between Rymovirus and Ipomovirus. The sequence comparison and phylogenetic studies revealed that the relatedness of SCSMV with the potyviral related genera was comparatively low to consider it as a member of earlier described potyviral genera, hence the genus “Susmovirus” (sugarcane streak mosaic virus) has been proposed, with SCSMV as the sole species to be included. The 52 SCSMV-CB isolates from this institute were distributed in 14 phylogenetic groups and the grouping pattern revealed that the virus isolates could not be grouped based on geographical origin of the host varieties or longevity of the host variety.     

Mycoflora associated with some freshwater plants collected from Delta region (Egypt)

77 species and 3 species varieties belonging to 35 genera were isolated during this investigation. Five species namely: Chytriomyces annulus, Rhinocladiella schulzeri, Dactylella bemicodes, D. alaskana and D. rhompospora are new records to Egypt. The composition of water-borne fungi was basically similar to those of the washwater of collected plants (phyllosphere) and plant part samples (phylloplane). Of the 20 zoosporic fungal species, Saprolegnia ferax and Dictyuchus sterilis were common in water samples; and Achlya oligacantha and S. ferax in washwater (phyllosphere). On glucose and cellulose-CZAPEK's agar the most frequent of the three substrates were the members of Fusarium, Aspergillus, Cladosporium, Alternaria and Penicillium. Also many more species such as Alternaria alternata, Cladosporium cladosporioides, Fusarium moniliforme and Stachybotrys atra increased their frequency of occurrence and total counts on cellulose agar than was the case on glucose. Some fungal species were isolated only from water samples and not from phyllosphere or phylloplane and vice versa. Also, some fungal species were isolated on glucose and not on cellulose CZAPEK's agar medium and vice versa. The plant Ceratophyllum demersum exhibited the highest fungal counts while Phragmites communis exhibited the lowest counts. The water samples richer in fungal genera and species were those which were characterized by relatively high content of dissolved oxygen. pH, temperature and conductivity of collected water samples did not show any regular pattern and did not exhibit any considerable influence on fungal population.     

Fungi from water,soil and air polluted by the industrial effluents of Manquabad superphosphate factory (Assiut,Egypt)

One hundred and one species in addition to three varieties appertaining to 53 genera of zoosporic (25 species and 15 genera) and terrestrial (76 species + 3 varieties and 38 genera) fungi were isolated during this investigation. The water and submerged mud directly exposed to industrial effluents were the poorest in terrestrial fungi whereas zoosporic fungi were completely missed. These samples were highly acidic, low content of oxygen and relatively high contents of total soluble salts, phosphate, sulphate, calcium and magnesium. Dictyuchus sterilis and Aphanomyces laevis were the most common zoosporic species using baiting technique whereas Aspergillus fumigatus, A. niger, A. flavus, Alternaria alternata, Gibberella fujikuroi and Stachybotrys chartarum were the most prevalent terrestrial fungal species using glucose- and cellulose-CZAPEK 's agar media. Tripospermum sp. (aquatic hyphomycetes) was isolated from the phyllosphere of Myriophyllum spicatum and is new record to Egypt. There are some variations concerning the fungal diversity frequencies and populations with regard to the test sources and media used.     

Seasonal fluctuations of aquatic fungi recovered from Egyptian soil (Delta Region)

One hundred and eleven identified and seven unidentified species in addition to 5 varieties belonging to forty-three aquatic fungal genera were recovered from 452 soil samples collected randomly from different localities in Nile Delta and Suez canal regions including eleven governorates. The investigated soil samples were collected seasonally during the period from autumn 1990 to summer 1991 (113 samples each season). The richest season (85 identified and 5 unidentified species in addition to 2 varieties) in aquatic fungi was the winter season (10.0-15.5°C) whereas the poorest (32 identified and 5 unidentified species in addition to one variety) was summer (24.0-27.5°C). The richest soil samples in aquatic fungi were those of low or moderate temperature seasons, high contents of organic matter and considerably poor in total soluble salts. The pH value did not show any regular seasonal variation and did not exhibit any considerable influence on fungal populations. Some aquatic fungal genera and species were disappeared completely in certain seasons and vice versa. The most dominant aquatic fungal genera were Pythium, Allomyces, Aqualinderella and Nowa-kowskiella. The commonest aquatic fungal species were Allomyces anomalus, Aqualinderella fermentans, Nowakowskiella elegans, Blastocladiopsis parva and Dictyuchus sterilis.     

Effects of Hericium erinaceus Hedgehog mushroom on the endophytic microbial community of the host plant

Hedgehog mushroom is a Hericium erinaceus associated with fagaceae and pinaceae trees in the northern hemisphere. It is still unknown whether this symbiotic relationship will affect the endophytic microbial community of the host plants. In this study, the endophytic microbial communities of different Quercus aliena tissues (root, stem, and leaf) with or without H. erinaceus partner were analyzed by bar-coded pyrosequencing. About 29,000 clean reads were obtained per sample representing 28 phyla of bacteria and 6 phyla of fungi. A total of 26,838 operational taxonomic units (OTUs) of bacteria and 4323 OTUs of fungi were observed at a 97% similarity level. Three bacterial phyla, Proteobacteria, Cyanobacteria and Bacteroidetes, and fungal phylum Ascomycota were dominant in all tissues. The relative abundance of these dominant communities showed significantly differences between Q. aliena tissues with or without H. erinaceus. Bacterial genus Pseudomonas and fungal genus Cryptosporiopsis were species-rich in Q. aliena root infected by H. erinaceus hyphae. This study demonstrated that the endophytic microbial community structure and dominant species varied in Q. aliena mycorrhized with H. erinaceus.     

Keratinophilic fungi from the feathers of Emu (Dromaius novaehollandiae) in Maharastra India

A total of 70 feathers samples of Emu (Dromaius novaehollandiae) were collected from 7 Emu farms situated at two districts (Raigad and Thane) of Maharashtra (India) and screened for resident keratinophilic fungi. Among them, 44 isolates were recovered and identified by evaluating characteristic macro- and micro-morphological features. Further gene products corresponding to the ITS1-5.8S-ITS2 rDNA region from all isolates were amplified and sequenced. Homology search was performed using BLAST program against non-redundant nucleotide database, and significantly matched DNA sequences deposited to the NCBI Gene Bank for reference purposes. Eight identified fungal species belongs to 7 different genera named as Aphanoascus terreus Ac_MW577456 (21.43%), Microsporum gypseum Ac_MW580920 (14.29%), Ctenomyces serratus Ac_MW577459 (10.0%), Uncinocarpus orissi Ac_MW577461 (5.17%), Aphanoascus verrucosus Ac_MW577458 (4.29%), Gymnascella dankaliensis Ac_MW577460 (2.86%), Gymnoascoideus petalosporus Ac_MW577462 (2.86%) and Arthroderma tuberculatum Ac_MW577457 (1.43%).     

Pathogenicity of some hypocrealean fungi to adult Aedes aegypti (Diptera: Culicidae)

The pathogenicity of 19 hypocrealean entomopathogenic fungi from seven different genera in adult Aedes aegypti was tested. All fungi proved to be pathogenic, and Isaria fumosorosea, Lecanicillium muscarium, Lecanicillium psalliotae, Metarhizium anisopliae, Metarhizium lepidiotae, Metarhizium majus, Metarhizium frigidum, Paecilomyces carneus, and Paecilomyces lilacinus caused total mortality within 15 days of exposure of mosquitoes to the fungal culture. All fungi developed on dead individuals. The high susceptibility of adults to most tested strains underlines the interest of entomopathogenic fungi—especially those of the genera Metarhizium, Isaria, Paecilomyces and Lecanicillium—for biological control of A. aegypti.     

Immunobiochemical Analysis of Cross-Reactive Glutathione-S-Transferase Allergen from Different Fungal Sources

Clinical observations suggest the presence of cross-reactive allergens. There is a need to identify these cross-reactive allergens to improve the treatment used for allergic disorders. The present study was aimed to identify and characterize a cross-reactive allergenic protein from fungi. Allergen extracts of various fungi viz. Alternaria alternata, Aspergillus fumigatus, Cladosporium herbarum, Curvularia lunata, and Epicoccum purpurascens showed GST enzymatic activity ranging from 0.765 to 1.004 Δ340 nm/min/µg where as activity of rGST was 1.123 Δ340 nm/min/µg. Immunoblot with GST antibodies showed a band of approximately 26 kDa in all these fungal extracts. Sera of fungal allergy patients showed the presence of IgE antibodies to GST. Rabbit antibodies raised against the fungal extracts reacted with rGST confirming the presence of GST-like protein in these extract. ELISA inhibition using GST antibodies revealed inhibition with C. herbarum, A. alternata, C. lunata, A. fumigatus, and E. purpurascens demonstrating that fungal GST competes for binding to anti-GST. In summary, a GST-like protein was recognized as cross-reactive allergen in these fungal extracts.     

Endophytic fungi with antitumor activities: Their occurrence and anticancer compounds

Abstract

Plant endophytic fungi have been recognized as an important and novel resource of natural bioactive products, especially in anticancer application. This review mainly deals with the research progress on the production of anticancer compounds by endophytic fungi between 1990 and 2013. Anticancer activity is generally associated with the cytotoxicity of the compounds present in the endophytic fungi. All strains of endophytes producing antitumor chemicals were classified taxonomically and the genera of Pestalotiopsis and Aspergillus as well as the taxol producing endophytes were focused on. Classification of endophytic fungi producing antitumor compounds has received more attention from mycologists, and it can also lead to the discovery of novel compounds with antitumor activity due to phylogenetic relationships. In this review, the structures of the anticancer compounds isolated from the newly reported endophytes between 2010 and 2013 are discussed including strategies for the efficient production of the desired compounds. The purpose of this review is to provide new directions in endophytic fungi research including integrated information relating to its anticancer compounds.     

Genetic transformation of Diaporthe phaseolorum, an endophytic fungus found in mangrove forests, mediated by Agrobacterium tumefaciens

We describe the genetic transformation of the mycelial tissue of Diaporthe phaseolorum, an endophytic fungus isolated from the mangrove species Laguncularia racemosa, using Agrobacterium tumefaciens-mediated transformation (ATMT). ATMT uses both the hygromycin B resistant (hph) gene and green fluorescent protein as the selection agents. The T-DNA integration into the fungal genome was assessed by both PCR and Southern blotting. All transformants examined were mitotically stable. An analysis of the T-DNA flanking sequences by thermal asymmetric interlaced PCR (TAIL-PCR) demonstrated that the disrupted genes in the transformants had similarities with conserved domains in proteins involved in antibiotic biosynthesis pathways. A library of 520 transformants was generated, and 31 of these transformants had no antibiotic activity against Staphylococcus aureus, an important human pathogen. The protocol described here, using ATMT in D. phaseolorum, will be useful for the identification and analysis of fungal genes controlling pathogenicity and antibiotic pathways. Moreover, this protocol may be used as a reference for other species in the Diaporthe genus. This is the first report to describe Agrobacterium-mediated transformation of D. phaseolorum as a tool for insertional mutagenesis.     

Studies on air-borne fungi at Qena I. Seasonal fluctuations

73 species which belong to 24 genera were collected in 200 and 35 esposures made during the period May 1976–October 1977 at each of two levels (2 m and 20 m). The air-borne fungi showed seasonal periodicities and the highest incidence was recovered in spring and autumn and the least in summer. Aspergillus was the dominating genus. 17 species were collected at the two levels of which A. niger and A. flavus were the most common. Other common genera were Cladosporium which was represented by C. herbarum, C. cladosporioides, C. sphaerospermum, and C. macrocarpum. 7 species of Curvularia were identified of which C. pallescens was the most frequent at the low and C. spicifera at the high level. Drechslera was represented by 6 species of which D. halodes was the most common at the two levels. Only one Alternaria species, A. alternata was isolated at both levels. 10 Penicillium were recovered, P. notatum was the most frequently one isolated at the two levels. Many fungal spore showers of Cladosporium, Aspergillus, Curvularia, and Alternaria were recorded during the experimental period.     

Increased prevalence and altered species composition of filamentous fungi in respiratory specimens from cystic fibrosis patients

Filamentous fungi cultured from respiratory tract specimens submitted to the department of clinical microbiology, Aarhus University Hospital, during 2010 were identified by morphology and by internal transcribed spacer (ITS) sequencing. Of 343 fungal isolates, discrepancies between identification methods were observed for four isolates (1.2%), while identification to species was achieved only with ITS sequencing for 16 isolates (4.7%). Filamentous fungi were isolated from 15% of cystic fibrosis (CF) respiratory samples in contrast to 2% of non‐CF samples. From CF patients, a total of nine different species were found in 188 samples from 48 patients, whereas from non‐CF patients, 24 different species were found in 155 samples from 111 patients. CF was associated with a significant overrepresentation of Aspergillus fumigatus and Scedosporium species; in contrast, the frequency of Penicillium spp. and other putative contaminants were significantly increased in non‐CF patients. The altered species variation of filamentous fungi in CF respiratory specimens is contradictory to a scenario of incidentally inhaled spores, trapped in the viscous airway mucus of these patients and subsequently expectorated; rather, our data most likely reflect both an increased prevalence and an increased proportion of truly colonizing fungi in this patient group.     

Naphthalene-anthracene-utilizing microorganisms

Oil-polluted freshwater samples collected from Assiut oil company, Egypt, were screened for hydrocarbon-utilizing microorganisms (bacteria and fungi). Two hydrocarbons (naphthalene and anthracene) were used as a sole source of carbon. Most Probable Number (MPN) technique was used to determine naphthalene and anthraceneutilizing bacteria. Micrococcus, Corynebacterium, Nocardia and Pseudomonas were recovered on basal medium supplemented with the hydrocarbons. Numbers of bacteria decreased as the pollution in water increased. Eleven species and 1 variety belonging to 8 terrestrial fungal genera were collected from oil polluted water on naphthalene or anthracene modified CZAPEK agar medium. Aspergillus (A. flavus var. columnaris) and Scopulariopsis (S. brumptii) were the most prevalent genera. 75.4 and 24.6% of total bacterial isolates utilized naphthalene and anthracene with different ranges of turbidity, respectively.