Endophytic Fungal Assemblages from Inner Bark and Twig of Terminalia arjuna W. & A. (Combretaceae)

Summary Fungal endophytes reside in healthy tissues of all terrestrial plant taxa studied to date and are diverse and abundant in tropical woody angiosperms. Endophytic fungi were isolated from Terminalia arjuna, an important ethno pharmacological plant extensively used in ayurvedic medicines to treat heart ailments. Isolations were made from symptomless fresh inner bark as well as twig samples of five plants collected from three locations of riparian vegetation during two seasons (monsoon and winter) of 2003 and 2004. Two hundred and seventy eight isolates, representing 22 genera, were obtained from both seasons. Monsoon seasonal isolations representing 22 genera showed greater diversity. Coelomycetes were more numerous during the winter season than hyphomycetes and ascomycetes. Among the endophytes, the genus Pestalotiopsis dominated the endophyte assemblage of T. arjuna collected from different locations, dominance was greater during the winter season than the monsoon season. Endophytic colonization frequency was greater in inner bark (18.5%) than twigs (4.6%). The genera Pestalotiopsis (54.5%), Chaetomium (10.5%) and Myrothecium (9%) were the most predominant endophytes. Rarefaction indices indicated the highest expected number of species for bark samples, monsoon isolations and location 1 (Mysore).     

Three new Lasiodiplodia spp. from the tropics, recognized based on DNA sequence comparisons and morphology

Botryosphaeria rhodina (anamorph Lasiodiplodia theobromae) is a common endophyte and opportunistic pathogen on more than 500 tree species in the tropics and subtropics. During routine disease surveys of plantations in Australia and Venezuela several isolates differing from L. theobromae were identified and subsequently characterized based upon morphology and ITS and EF1-alpha nucleotide sequences. These isolates grouped into three strongly supported clades related to but different from the known taxa, B. rhodina and L. gonubiensis, These have been described here as three new species L. venezuelensis sp. nov., L. crassispora sp. nov. and L. rubropurpurea sp. nov. The three could be distinguished easily from each other and the two described species of Lasiodiplodia, thus confirming phylogenetic separations. Furthermore all five Lasiodiplodia spp. now recognized separated from Diplodia spp. and Dothiorella spp. with 100% bootstrap support.     

Study on the diversity of endophytic communities from rice (Oryza sativa L.) and their antagonistic activities in vitro

Endophytic populations were isolated from 2400 segments of Oryza sativa collected from Bhadra River Project Area, Southern India during December 2005 (Winter) and April 2006 (Summer). Overall colonization rates from surface sterilized tissues were 40.3% in roots and 25.83% in leaves during winter season, 20.15% in roots and 8.66% in leaves during summer season. Nineteen different fungal taxa, a Streptomyces sp. and bacterial species were isolated. Streptomyces sp., Chaetomium globosum, Penicillium chrysogenum, Fusarium oxysporum and Cladosporium cladosporioides were dominant endophytes in this study. Frequency of colonization between the sites, seasons and rice varieties were found to differ significantly. Dual culture studies revealed that C. globosum, P. chrysogenum and Streptomyces sp. are suitable candidates for extraction of biologically active compounds. Rice harbors many endophytic organisms and some of them have antagonistic properties against fungal pathogens.     

Endophytic fungi from Amomum siamense.

Endophytic fungi were isolated from apparently healthy organs of the wild ginger Amomum siamense Criab.. including leaves, pseudostems, and rhizomes, collected from two sites in Doi Suthep-Pui National Park, Thailand. Endophytes were relatively common with an isolate prevalence of 70%-83% at the two sites sampled in the wet and dry seasons. The endophyte assemblages from the two sites were diverse and comprised 7 Ascomycetes and 26 mitosporic fungi. Colletotrichum "gloeosporioides" (Penz.) Penz. & Sacc. in Penz., Glomerella spp., xylariaceous fungi, and Phomopsis spp. were consistently dominant as endophytes on Amomum siamense, but many rare species and mycelia sterilia were also recorded. There was no significant difference between the number of isolates recovered from leaves containing vein tissues and those containing intervein tissues, independent of leaf age. Most taxa showed a preference for either leaf tissue or pseudostems. Two new Ascomycetes species. Gaeumannomyces amomi and Leiosphaerella amomi, were discovered from leaves and rhizomes, respectively, and four species of Pyricularia, including three new species, were isolated from leaves.     

Endoglucanase production by paper-degrading mycoflora

Fourteen fungal species, namely Aspergillus flavus, A. fumigatus, A. niger, A. ustus, Penicillium islandicum, P. wortmannii, Memnoniella echinata, Cladosporium herbarum, Stachybotrys atra, Chaetomium globosum, Fusarium oxysporum, Torula herbarum, Alternaria alternata and Curvularia uncinata were isolated from different grades of paper. They differ in their distribution on various kinds of paper and also in relative occurrence. While seasonal influence on mycoflora was observed, most of the moulds were capable of growing in all three seasons examined (summer, winter, rainy season). The moulds were cellulolytic in nature and endoglucanase activity was greatest in Aspergillus flavus, A. niger, A. fumigatus, P. wortmannii and P. islandicum.     

Diversity of fungal endophytes in shrubby medicinal plants of Malnad region,Western Ghats,Southern India

A total of 6125 fungal endophytes were isolated from 9000 leaf segments of 15 medicinal shrubs growing in Malnad region of Western Ghats, Southern India, during winter, monsoon, and summer seasons. These fungal isolates belonged to Ascomycota (8.6 %), Coelomycetes (26.0 %), Hyphomycetes (28.0 %), Mucoromycotina (0.3 %) and sterile forms (4.9 %). Alternaria, Chaetomium, Fusarium, Colletotrichum, Cladosporium, Penicillium, Phyllosticta and Xylaria were the most frequently isolated. Significantly more isolates were obtained during the winter season than monsoon and summer seasons.     

Limnological studies on freshwater ponds of Hyderabad — India

Seasonal abundance of phytoplankton has been discussed in three tropical ponds of Hyderabad, India during two years of investigation. The observed patterns have been characterized as seasonal maxima and minima during summer, monsoon and winter. The tropical patterns observed in the study ponds were typical but different from those observed in temperate regions. Most of the maxima were observed in winter and the minima during the monsoon or rainy season. Diatomeae dominated the phytoplankton of all the ponds in all seasons. An inverse relationship between Diatomeae and Cyanophyta, and Diatomeae and Euglenineae was observed. The phytoplankters have been grouped as perennial, summer, monsoon and winter species.     

Diversity and antimicrobial activities of the fungal endophyte community associated with the traditional Brazilian medicinal plant Solanum cernuum Vell. (Solanaceae)

The diversity and antimicrobial activity of endophytic fungi associated with the Brazilian medicinal plant Solanum cernuum Vell. were studied during summer and winter seasons. A total of 246 fungal isolates were obtained, including 225 filamentous fungi and 21 yeasts. They were identified by morphological, physiological, and molecular methods. Fifty-five different taxa represented by the phyla Ascomycota (33 taxa), Basidiomycota (21 taxa), and Zygomycota (one taxon) were identified. The most abundant taxa were closely related to Arthrobotrys foliicola , Colletotrichum gloeosporioides , Coprinellus radians , Glomerella acutata , Diatrypella frostii , Phoma glomerata , Mucor sp., Phlebia subserialis , Phoma moricola , Phanerochaete sordida , and Colletotrichum sp. A total of 265 fungal extracts were screened and 64 (26.01%) displayed antimicrobial activities. Among these extracts, 18 (28.12%) presented antibacterial and antifungal activities, 42 (65.62%) displayed selective antibacterial activity, and four (6.25%) exhibited only antifungal activity. The best values of minimum inhibitory concentration were obtained from extracts of Cryptococcus rajasthanensis , Glomerella acutata, Leptosphaeria sp., and Phoma glomerata ranging from 7.8 to 15.62?μg/mL. This study is the first survey of the endophytic fungi community associated with S. cernuum, and our results show that they can represent a promising source of bioactive compounds.     

Comparative effect of live food and manured treatments on water quality and production of ornamental carp, Cyprinus carpio var. koi L., during winter, summer, monsoon and post-monsoon growout experiments in concrete tanks

Four growth trials (each of 11 weeks duration) were conducted during different seasons throughout 1 year, namely, winter, summer, monsoon and post‐monsoon to assess the seasonal influence on growth and production of koi carp, Cyprinus carpio var. koi L. larvae in concrete tanks maintained under different management regimes. Individual weight gain, survival rate, fish deformities and number of marketable fish in each season were compared among four management regimes: (i) carp larvae fed with live zooplankton (LF); (ii) direct fertilization with poultry manure (PM); (iii) direct fertilization with cow dung (CD); and (iv) a control system (C), where a commercial diet containing 32% crude protein was applied. There were three replicates for each treatment. The LF treatment produced significantly higher weight gain, survival rate and number of marketable fish in all seasons (P < 0.05), compared with the other treatments, through maintenance of better water quality and greater abundance of zooplankton in the system. Fish deformities were highest in the C treatment in every season (P < 0.05). Water temperatures averaged 18.6°C (winter), 29.7°C (summer), 28.3°C (monsoon), and 26.5°C in the post‐monsoon trial. Average weight gain of koi carp achieved during the winter trial (2.01 g in C to 4.44 g in LF) was considerably lower than that achieved in the summer, monsoon or post‐monsoon trials. Survival rates of carp ranged from 70.5% (C) to 95.5% (LF) in the winter trial, and were considerably lower than the other seasons. The number of marketable fish was also lowest during the winter trial.     

Assessment of diversity,distribution and antibacterial activity of endophytic fungi isolated from a medicinal plant <Emphasis Type="Italic">Adenocalymma alliaceum</Emphasis> Miers

A study was conducted for isolation, identification and antibacterial potential of fungal endophytes of Adenocalymma alliaceum Miers., (Bignoniaceae), a medicinal shrub vine plant which has long history for its usages in curing various disorders. A total of 149 isolates of endophytic fungi representing 17 fungal taxa were obtained from 270 segments (90 from each stem, leaf and petiole) of this plant. Hyphomycetes (77.85%) were the most prevalent, followed by Ascomycetes (8.05%) and Coelomycetes (4.03%) respectively. A considerable amount of fungal isolates was kept under (10.07%) Mycelia-Sterilia (MS). Leaf harboured maximum colonization of endophytic fungi (72.22%) which was greater than stem (67.78%) and petiole (25.54%). The Jc similarity index was maximum (0.619) between stem vs leaf followed by leaf vs petiole (0.571) and stem vs petiole (0.428). The dominant endophytic fungi were Alternaria alternata, Aspergillus niger, Stenella agalis, Fusarium oxysporum, Curvularia lunata and Fusarium roseum. Among twelve endophytic fungi tested for antibacterial activity, crude extracts of nine endophytic fungi (75%), showed antibacterial potential against one or more clinical human pathogens. Alternaria alternata, Curvularia lunata, Penicillium sp. and Chaetomium globosum exhibited significant antibacterial activity against 4 of 5 tested pathogens, showing broad spectrum activity. This investigation explains the value of sampling from different tissues of a host plant for the greater species diversity, and additionally, the antibacterial screening of some endophytic fungi from this specific medicinal plant may represent a unique source for many of the useful antibacterial compounds.     

Seabirds attending bottom long-line fishing off southeastern Brazil

Flocks of seabirds attending commercial bottom long-line fishing operations on the coastal shelf off southeastern Brazil show a greater species diversity during the summer than the winter (16 v 9 species), although the number of birds per flock tended to be greater during the cold season. During the summer, the Spectacled Petrel Procellaria aequinoctialis conspi-cillata was the commonest species, followed by the Great Shearwater Puffinus gravis, Cory's Shearwater Calonectris diomedea, skuas Stercorarius spp. and the Yellow-nosed Albatross Diomedea chlororhynchus. During the winter, the White-chinned Petrel Procellaria aequin-octialis, Yellow-nosed Albatross and Black-browed Albatross Diomedea melanophrys were jointly the commonest species. Marked differences in the relative abundance of species were observed between the different sampling periods, probably because of migratory movements but also because of seasonal shifts of the sea currents and the influence of cold fronts. Waters off southeastern Brazil are important feeding areas for some seabird populations nesting in the Tristan da Cunha and Gough group, especially for nonbreeding Spectacled Petrels and post-breeding Yellow-nosed Albatrosses.     

Investigations into the soil mycoecology of Chambal ravines of India

Summary The present investigation deals with the seasonal succession of fungal communities in wheat fields. The studies were performed during the year 1975–76 and involved four study seasons,viz. winter (1975), summer (1975), rainy (1975) and winter (1976). The observations during two winter seasons were done to find out whether or not fungal communities exhibit definite cyclic pattern. Of the 69 isolated species, 25 showed wider ecological amplitude whereas some species were specific of a particular season. Each season was marked by different fungal mycoflora. The population ofAspergillus terreus, a dominant member of the community, along with many other autochthonous species, exhibited a definite periodicity during various seasons. Seasonal variations in the fungal communities, variations in the extent of similarity among the fungal communities during various seasons and the impact of climatic conditions on the components of fungal communities are discussed.     

Antimicrobial endophytic fungal assemblages inhabiting bark of <Emphasis Type="Italic">Taxus baccata</Emphasis> L. of Indo-Burma mega biodiversity hotspot

Fungal endophytes were isolated from inner bark of Taxus baccata L., an important source of potent anticancer drug taxol. Bark samples were collected from two locations of Arunachal Pradesh, India, part of the Indo-Burma mega biodiversity hotspot, during two seasons i.e. monsoon and winter. Altogether 77 fungal strains representing 18 genera were isolated from T. baccata bark during the present investigation. The colonizing frequency was recorded as 38.5% and the fungal community comprised of 78% of Hyphomycetes, 5.2% of Coelomycetes, 2.6% of Zygomycetes and Ascomycetes and 9.1% of sterile mycelia. Most common and frequently isolated genera were Fusarium, Penicillium and Aspergillus. Simpson and Shannon diversity indices indicated higher species diversity during monsoon than during winter seasons irrespective of the locations. The two locations harbored 5 to 37 endophyte species and the similarity index was low during winter and high during monsoon. Ethyl acetate extract of fermentation cultures of these fungi were tested for their antimicrobial activity against a panel of human pathogenic Gram-positive and Gram-negative bacteria and fungi. Fifteen fungal isolates out of the isolated strains displayed antimicrobial activity. An endophytic fungus, identified as Fusarium sp. displayed significant antimicrobial activity against all the test pathogens.     

Effect of seasonal climatic changes on biomass yield and terpenoid composition of rose-scented geranium (Pelargonium species)

Variations in biomass yield, essential oil yield and terpenoid composition in rose-scented geranium (Pelargonium species) in response to seasonal climatic changes were investigated under semiarid tropical climatic conditions. A large number of essential oil samples were collected during different seasonal months (once a month) and daily during the peak summer season months of May and June. They were analysed for terpenoid composition by GC and GC-MS. The crop yielded the lowest values for biomass yield, essential oil yield and essential oil concentration in the summer months of April to June. Evaluation of terpenoid compositions showed minimum concentrations (% of essential oil) of linalool, geraniol and its esters and maximum concentrations of citronellol and its esters during summer months. The percentages of geraniol and its esters were highest during cool winter season months of December and January followed by rainy and autumn season months. Isomenthone, 10-epi-gamma-eudesmol and other minor terpenoid compounds (present in <1% amounts in the essential oil) did not exhibit any definite seasonal trends. Rainy/monsoon (August and September) and autumn (October and November) season months were characterised by high rainfall, cloudy days and short photoperiods. These favourable environmental conditions encouraged crop growth and produced highest biomass yields, essential oil yields and maximum concentration of essential oil in rose-scented geranium plants.     

Seasonal variation in the activity of the Leydig cells in Egyptian Nubian goat (Zaraibi) bucks

Ten Egyptian Nubian goat bucks were used to evaluate the effect of season on testicular hormonal activity and ultrastructure. Parameters were recorded for 7 consecutive weeks in the middle of the four seasons, with blood samples being collected weekly. At the end of each of these seasons, testicular biopsies were obtained surgically for histological and cytological studies. Season had a significant effect on plasma testosterone concentration, being at its lowest level (P < 0.01) during winter and spring (1.2 and 2.6 ng/ml, respectively), while at its highest during summer (10 ng/ml). The effect of season on plasma LH concentration was higher (P < 0.01) in autumn (2.9 mIU/ml) and less in spring and summer (0.4 mIU/ml). Season of the year influenced the percentage of sectional tissue area occupied by the seminiferous tubules and interstitial tissue. Seminiferous tubules occupied the majority of the testicular tissue during winter (76.6%), with the least being occupied during spring (49.8%). The thickness of the seminiferous tubules was maximal during autumn, followed by summer (53 and 36 μm, respectively). In summer the Leydig cells contained abundant smooth endoplasmic reticulum (sER), while some areas of the cytoplasm were occupied exclusively by tubular sER, arranged in parallel—indicating the highest activity of these cells. A characteristic multivesicular structure with numerous large lipid droplets and vacuoles was recorded in the Leydig cells during spring and winter, denoting low or even arrested activity of the cells. It could be concluded that season influences the activity of the Leydig cells of Egyptian Nubian bucks, and this is reflected by their ultrastructure and secretive activity.     

Endophytic fungal diversity of 2 sand dune wild legumes from the southwest coast of India

Endophytic fungi of 3 age classes (seeds, seedlings, and mature plants) and 5 tissue classes (cotyledons, seed coats, roots, stems, and leaves) of coastal sand dune legumes Canavalia cathartica and Canavalia maritima were assessed by plating surface-sterilized segments on malt extract agar. Forty-six fungal taxa comprising 6 ascomycetes, 33 mitosporic fungi, 2 zygomycetes, and 5 sterile morphospecies were recovered. There was no significant difference in the colonization frequency of endophytes between plant species (p = 0.4098, Student's t test). Among the age classes, endophytic fungi colonized over 90% of seedlings and mature plants. Similarly, among tissue classes, endophytic fungi colonized over 90% of root, stem, and leaf segments. Diversity and richness of endophytic fungi were higher in C. cathartica than in C. maritima. Rarefaction curves revealed a "higher expected number of species" in mature plants of C. cathartica and seedlings of C. maritima, whereas it was highest in leaves of both plant species. The most dominant endophyte, Chaetomium globosum, colonized over 50% of the root, stem, and leaf segments of C. maritima and over 50% of the root segments of C. cathartica. The colonization frequency of C. globosum was found to be 5%-12.5% in seeds and increased up to 40%-64.4% in seedlings or mature plants. Halosarpheia sp. was the only marine fungus recovered among the endophytes.     

Emerging new crown symptoms on Castanea sativa (Mill.): Attempting to model interactions among pests and fungal pathogens

In the 2015–2016 growing seasons, two novel symptoms were assessed on the crown of trees in orchards and coppices of chestnut groves in Central Italy. The first symptom was flagging of annual shoots with green leaves undergoing sudden wilt and turning brown later in the season. The second symptom consisted of leaves on annual shoots turning yellow before wilting in absence of flagging represented the second symptom. Samples were collected along transects in early summer, late summer and winter, and processed in the laboratory. The flagging symptom was associated in early summer with the presence of C. parasitica in cryptic dried buds on stems from the previous year's growth. The pathogen was also found in dormant buds in winter, suggesting that the infection could take place in summer during the Chinese gall wasp oviposition period. Cryphonectria parasitica was also isolated from abandoned galls in winter supporting the hypothesis that galls are a potential source of inoculum for crown infections. Aetiology of yellowing was not clarified and no fungal taxa were specifically associated with this symptom. Gnomoniopsis castanea, C. parasitica and, in early summer, Colletotrichum acutatum were the most abundant fungal taxa isolated from chestnut shoots and buds.     

Planktonic larval stages of Brachyura in the Gulf of Tigullio (Ligurian Sea,Italy)

Summary

Horizontal planktonic catches made in the Gulf of Tigullio (Genoa, Ligurian Sea) have been analyzed with the aim of identifying the zoeae of Brachyura, whose knowledge is poor in all the northern Mediterranean Sea. Forty-four taxa (42 species plus Liocarcinus spp. and Brachynotus spp.) have been identified, 12 of which were present with all the larval stages. The seasonal distribution of the taxa has been analyzed, and it was possible to recognize species present throughout the year as well as spring/summer, summer/autumn, and winter/summer species. Problems regarding zoeae of dubious identification are discussed.     

Colonization of experimentally disturbed patches by stream macroinvertebrates in the Acheron River,Victoria

Twenty-five patches (1 m²) of natural stream substratum in the Acheron River, Victoria, were physically disturbed by kicking and raking during winter 1986 and summer 1987. The macroinvertebrate composition of these disturbed patches was examined at various times over the following 71 days, and compared with adjacent undisturbed control patches sampled concurrently. The disturbance did not alter the particle-size distribution (> 150 μm) of the disturbed patches. Organic material was reduced in the disturbed patches by about 70% in each season, but returned to control levels within 21 days in winter and 8 days in summer. The total number of species, and the density of species and individuals were all significantly reduced by the disturbance. Recovery of species density was complete after 21 days during winter and 8 days during summer, and the density of individuals recovered after 71 days during winter and 8 days during summer. The differences were due to the slower colonization rate of Chironomidae in winter, either because of a lower drift rate, or a slower recovery of detritus in winter. Individual species showed variations in colonization patterns, most increasing steadily at various rates, with some declining after an initial rapid increase (e.g. Baetis pp.). In the latter case, the density changes were mirrored in the control patches, emphasizing the need to take control samples concurrently with experimental samples. In each season, the species remaining immediately following the disturbance, and those subsequently colonizing the disturbed patches were in the same rank order (Spearman Rank correlation) as their occurrence in the control patches, suggesting that no taxa were differentially affected by the treatment. No evidence was found to allow the application of the Intermediate Disturbance Hypothesis to explain species diversity at the scale of this study. It appears that current hypotheses developed to explain the relationship between diversity and disturbance in sessile communities do not apply to highly mobile communities in streams.     

Significance of Molecular Identification and Antifungal Susceptibility of Clinically Significant Yeasts and Moulds in a Global Antifungal Surveillance Programme

The increasing diversity of opportunistic fungi causing serious invasive fungal infections (IFI) has been documented. Accurate identification (ID) is important in guiding therapy, determining prognosis for IFIs and in epidemiological surveys. We assessed the utility of PCR-based methods for the ID of yeasts and moulds that either were uncommon, failed conventional ID, or represented unusual biochemical or phenotypic profiles of common species. Among 1,790 viable fungal clinical isolates received during the SENTRY Program in 2010, 322 strains from 40 study sites had ID confirmed by molecular methods. Isolates were previously identified in participant institutions. Yeasts that were not confirmed by morphology on CHROMagar, growth at 45?°C (Candida albicans/dubliniensis), or assimilation of trehalose (C. glabrata) as well as non-Candida yeasts and all moulds were amplified and sequenced using primers amplifying one or more of the following genes: ITS, 28S, β-tubulin (Aspergillus spp.), TEF (Fusarium spp.), IGS (Trichosporon spp.). The isolates selected for molecular ID included 149 isolates of Candida species, 77 of Aspergillus species, 73 non-Candida yeasts, and 23 other moulds (a total of 41 different species). Overall, the ID determined by the submitting site was confirmed for 189 isolates (58.7?%): Aspergillus spp. (64.1?% correct); Candida spp. (60.1?% correct); non-Candida yeasts (58.9?% correct); non-Aspergillus moulds (30.4?% correct). Species with high levels of concordance between conventional and molecular ID included A. fumigatus (95.0 %), C. lusitaniae (100?%), C. dubliniensis (92.3?%), C. kefyr (100?%), and C. neoformans (90.2?%). Only 50.0?% of isolates of C. albicans and 59.1?% of C. glabrata selected due to unusual phenotypic or biochemical features were found to be correctly identified by the submitting site. Molecular methods for the identification of fungal pathogens are an important adjunct to the conventional identification of many less common clinically relevant yeasts and moulds including species of Candida with unusual or erroneous phenotypic or biochemical profiles. Molecular confirmation of fungal identification is essential in epidemiological surveys such as SENTRY.