DNA-based detection of the fungal pathogen Geomyces destructans in soils from bat hibernacula

White-nose syndrome (WNS) is an emerging disease causing unprecedented morbidity and mortality among bats in eastern North America. The disease is characterized by cutaneous infection of hibernating bats by the psychrophilic fungus Geomyces destructans. Detection of G. destructans in environments occupied by bats will be critical for WNS surveillance, management and characterization of the fungal lifecycle. We initiated an rRNA gene region-based molecular survey to characterize the distribution of G. destructans in soil samples collected from bat hibernacula in the eastern United States with an existing PCR test. Although this test did not specifically detect G. destructans in soil samples based on a presence/absence metric, it did favor amplification of DNA from putative Geomyces species. Cloning and sequencing of PCR products amplified from 24 soil samples revealed 74 unique sequence variants representing 12 clades. Clones with exact sequence matches to G. destructans were identified in three of 19 soil samples from hibernacula in states where WNS is known to occur. Geomyces destructans was not identified in an additional five samples collected outside the region where WNS has been documented. This study highlights the diversity of putative Geomyces spp. in soil from bat hibernacula and indicates that further research is needed to better define the taxonomy of this genus and to develop enhanced diagnostic tests for rapid and specific detection of G. destructans in environmental samples.     

Fungi isolated from Antarctic material

Summary Fungi isolated from samples of soil, penguin, skua and petrel dung and bird feathers in the Victoria Land, Antarctica, from Inexpressible Island to Cape King, were studied. All material was collected in December 1987–January 1988. Fungi occurred prevalently in bird dung and in soil, especially when mosses were present. The main species isolated were: the keratinophilic Chrysosporium verrucosum and Geomyces pannorum var. pannorum, Phoma herbarum and Thelebolus microsporus. A variety of filamentous fungi and yeasts were also encountered in soil, dung and bird feathers samples in different localities: Acremonium strictum, Cladosporium herbarum, Scolecobasidium salinum, Mortierella antarctica, Paecilomyces farinosus, Phialophora fastigiata, the thermophilic Scytalidium thermophile and Thermomyces lanuginosus, Verticillium sp., Mycelia sterilia and Cryptococcus albidus and Torulaspora delbrueckii. Most of the fungal isolates appeared to be cold-tolerant. Results from this study are discussed in conjuction with data from previous Antarctic studies in this area.     

嗜冷和冷育微生物的研究 Ⅲ.地丝霉属一新种,光滑地丝霉

在研究食品冷库中嗜冷微生物区系时,获得很多冷育型真菌。地丝霉和金孢菌类在这些菌中占优势。其中一菌在PYE上于18℃培养14天,菌落白色,厚毡状,中央常有淡黄色渗出液滴,菌落直径33mm,背面浅黄褐色,有辐射沟纹,老后仍为白色。菌丝分隔,直径1.0—2.0μm。该菌有明显的分生孢子梗,直立,宽度1.0—2.0μm,长10—25(—35)μm,无色或淡黄色,分生孢子梗具分枝,由分枝产分生孢子。分生孢子无色或淡黄色,顶端者为梨形、倒卵形或楔状,间生者矩椭圆形或桶状,1.5—2.5×2.5—3.5(—4.0)μm,光滑、1—3成链。该菌生长温度范围为0—25℃,最适温度为18℃,30℃不生长。此菌产生果胶酶,但是不分解角朊。该菌从冷库中生霉的蒜苗上分离得。其特征与Carmichael(1962),Domsch等(1980)所订正的金孢菌属(Chrysosporium Corda)中各种,以及Sigler和Carmichael(1976),Oorschot(1980)所订正的地丝霉属(Geomyces Traaen)中各种均有显著的差异,而其性状符合地丝霉属的特征集要。故命名为光滑地丝霉Geomyces laevis Z.Q.Li et Cui。模式:菌株AS 3.4605为模式菌株,保藏在中国科学院微生物研究所菌种保藏研究室;模式标本HMAS No.54875保存在该研究所真菌标本室。     

四川省土壤中的暗色丝孢菌

采用土壤平板法和稀释平板法,从采自四川省的24份土壤样品中,分离得到27个暗色丝孢菌分离物,隶属于16属24种。其中,浅色葡萄穗霉Stachybotrys pallescens为一新种,绿色暗梗单孢霉原变种Chloridium virescens var.virescens,大孢树粉孢原变种Oidiodendron maius var.maius和二色葡萄穗霉Stachybotrys dichroa为中国新记录。对新种及中国新记录作了较详细的描述并绘图。其余20个国内已报道种亦作了生境和分布地点的引证。所有研究菌株的干制培养物标本与活菌种均存放在山东农业大学植物病理学标本室(HSAUP)。     

Soil microfungi in three Swedish coniferous forests

Microfungi were isolated with the soil washing technique from five different soil horizons in three podzolic coniferous forests in Sweden. Totally 126 species were identified of which 45 were found at two and 24 were found at three sites. The most common genera were Mortierella and Penicillium . Other genera isolated in high frequences were Oidiodendron, Tolypocladium, Trichoderma and Verticillium . Pronounced differences in species composition were found between the soil layers, while great similarities were shown in identical soil layers at different sites.     

Aerial Transport of Keratinaceous Substrate and Distribution of the Fungus Geomyces pannorum in Antarctic Soils

Abstract Aerobiological sampling was conducted year-round at three sites on Signy Island, maritime Antarctic, between December 1992 and January 1994, and during the summer at a fourth coastal site. The concentrations of keratinaceous particles were monitored. Feather fragments were the most abundant of all particle types trapped. Seal hairs were also common, particularly on the coast. Numbers of both particle types were most common in the air in summer. The Antarctic soil fungus Geomyces pannorum, which is able to utilize keratin-based substrates, was also present in aerobiological samples on Signy Island. G. pannorum was only found during early winter on the year-round sampling sites. It was more abundant during summer in the air at the coastal site, which was more heavily influenced by seals, birds and humans. Skuas were swabbed when they returned to the island early in summer. G. pannorum was recovered in culture, suggesting that these birds may act as vectors for the transport of microorganisms between Antarctica and more northern landmasses. Organic material deposited on snow was concentrated on the soil at the edge of the ice cap by melt wash, making additional keratin and other organic substrates available to soil microorganisms. Received: 4 January 1998; Accepted: 25 February 1998     

Antarctic nematode communities: observed and predicted responses to climate change

The rapidly changing climate in Antarctica is impacting the ecosystems. Since records began, climate changes have varied considerably throughout Antarctica with both positive and negative trends in temperatures and precipitation observed locally. However, over the course of this century a more directional increase in both temperature and precipitation is expected to occur throughout Antarctica. The soil communities of Antarctica are considered simple with most organisms existing at the edge of their physiological capabilities. Therefore, Antarctic soil communities are expected to be particularly sensitive to climate changes. However, a review of the current literature reveals that studies investigating the impact of climate change on soil communities, and in particular nematode communities, in Antarctica are very limited. Of the few studies focusing on Antarctic nematode communities, long-term monitoring has shown that nematode communities respond to changes in local climate trends as well as extreme (or pulse) events. These results are supported by in situ experiments, which show that nematode communities respond to both temperature and soil moisture manipulations. We conclude that the predicted climate changes are likely to exert a strong influence on nematode communities throughout Antarctica and will generally lead to increasing abundance, species richness, and food web complexity, although the opposite may occur locally. The degree to which local communities respond will depend on current conditions, i.e., average temperatures, soil moisture availability, vegetation or more importantly the lack thereof, and the local species pool in combination with the potential for new species to colonize.     

Fungi are the predominant micro-organisms responsible for degradation of soil-buried polyester polyurethane over a range of soil water holding capacities

AIMS: To investigate the relationship between soil water holding capacity (WHC) and biodegradation of polyester polyurethane (PU) and to quantify and identify the predominant degrading micro-organisms in the biofilms on plastic buried in soil. METHODS AND RESULTS: High numbers of both fungi and bacteria were recovered from biofilms on soil-buried dumb-bell-shaped pieces of polyester PU after 44 days at 15-100% WHC. The tensile strength of the polyester PU was reduced by up to 60% over 20-80% soil WHC, but no reduction occurred at 15, 90 or 100% soil WHC. A PU agar clearance assay indicated that fungi, but not bacteria were, the major degrading organisms in the biofilms on polyester PU and 10-30% of all the isolated fungi were able to degrade polyester PU in this assay. A 5.8S rDNA sequencing identified 13 strains of fungi representing the three major colony morphology types responsible for PU degradation. Sequence homology matches identified these strains as Nectria gliocladioides (five strains), Penicillium ochrochloron (one strain) and Geomyces pannorum (seven strains). Geomyces pannorum was the predominant organism in the biofilms comprising 22-100% of the viable polyester PU degrading fungi. CONCLUSIONS: Polyester PU degradation was optimum under a wide range of soil WHC and the predominant degrading organisms were fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: By identifying the predominant degrading fungi in soil and studying the optimum WHC conditions for degradation of PU it allows us to better understand how plastics are broken down in the environment such as in landfill sites.     

Studies of pathogenic and antagonistic microfungal populations and their potential interactions in the mycorrhizoplane of Norway spruce (Picea abies (L.) Karst.) and beech (Fagus sylvatica L.) on acidified and limed plots

Recent tree decline was hypothesized to be connected to root damage caused by soil acidification and increased frequency of pathogenic root colonizing fungi. The rhizoplane is constituted by the mycorrhizal sheath and a high diversity of microfungi, some of which are known to behave antagonistically against pathogens. Disturbance of the balance between pathogens and antagonists by soil acidification may endanger the health of tree roots. Liming may stabilize the interactions. The microfungal populations connected to the mycorrhizoplane of Norway spruce (Picea abies) and beech (Fagus sylvatica) were, therefore, investigated on experimental Norway spruce plots that had been treated with acidified water or were limed. Beech presented the original forest and was left untreated. Eight microfungal species known as either pathogenic or antagonistic, Trichoderma viride, T. hamatum, T. polysporum, Cylindrocarpon destructans, Sesquicillium candelabrum, Mycelium radicis atrovirens, Tolyplocladium geodes and Oidiodendron maius, were isolated from the mycorrhizoplanes and their abundance in the five different plots compared. Acidification enhanced the frequency of Mycelium radicis atrovirens and Oidiodendron maius but reduced Trichoderma viride. Liming promoted Sesquicillium candelabrum and Cylindrocarpon destructans. Detailed analysis of the population patterns indicated that changes in the frequency of a particular fungal species may not only be caused by shift of chemical soil factors but also by antagonistic interactions between the microfungi, thus reducing pathogenic attacks on rootlets.     

Extracellular enzymes produced by microorganisms isolated from maritime Antarctica

Antarctic environments can sustain a great diversity of well-adapted microorganisms known as psychrophiles or psychrotrophs. The potential of these microorganisms as a resource of enzymes able to maintain their activity and stability at low temperature for technological applications has stimulated interest in exploration and isolation of microbes from this extreme environment. Enzymes produced by these organisms have a considerable potential for technological applications because they are known to have higher enzymatic activities at lower temperatures than their mesophilic and thermophilic counterparts. A total of 518 Antarctic microorganisms, were isolated during Antarctic expeditions organized by the Instituto Antártico Uruguayo. Samples of particules suspended in air, ice, sea and freshwater, soil, sediment, bird and marine animal faeces, dead animals, algae, plants, rocks and microbial mats were collected from different sites in maritime Antarctica. We report enzymatic activities present in 161 microorganisms (120 bacteria, 31 yeasts and 10 filamentous fungi) isolated from these locations. Enzymatic performance was evaluated at 4 and 20°C. Most of yeasts and bacteria grew better at 20°C than at 4°C, however the opposite was observed with the fungi. Amylase, lipase and protease activities were frequently found in bacterial strains. Yeasts and fungal isolates typically exhibited lipase, celullase and gelatinase activities. Bacterial isolates with highest enzymatic activities were identified by 16S rDNA sequence analysis as Pseudomonas spp., Psychrobacter sp., Arthrobacter spp., Bacillus sp. and Carnobacterium sp. Yeasts and fungal strains, with multiple enzymatic activities, belonged to Cryptococcus victoriae, Trichosporon pullulans and Geomyces pannorum.     

Potential for bioremediation of fuel‐contaminated soil in Antarctica

A preliminary investigation was conducted to identify the presence of bacteria in fuel‐contaminated Antarctic soil that could potentially be used to bioremediate the contaminated soil at McMurdo Station and other sites in Antarctica. The ability of soil microorganisms to metabolize fuels under the extreme climatic and oligotrophic conditions of Antarctica was of concern. Bacteria were isolated from fuel‐contaminated soil on site at McMurdo Station. Bacteria from noncontaminated soil near the station were also studied for comparison. The Antarctic soil microorganisms exhibited the ability to endure cold and oligotrophic environments. Experiments also showed that bacteria from the fuel spill site were active in their contaminated environment and that acclimation to xenobiotic compounds was necessary. Application of bioremediation in the extreme environmental conditions found at McMurdo Station, Antarctica, were also considered. The possibility of altering environmental factors necessary to adequately support in situ bioremediation in this extreme climate is discussed.     

Physiological diversity of bacterial communities from different soil locations on Livingston Island, South Shetland archipelago, Antarctica

Terrestrial food webs of Antarctica are simple and dominated by microorganisms. Soil bacteria play an important role in nutrient cycling, yet little is known about their capacity to utilize different carbon sources and to participate in site nutrient turnover. Biolog EcoPlate? was applied to study the catabolic activity and physiological diversity of bacteria inhabiting the soil of moss, vascular plants, and fell field habitats from Livingston Island, Antarctica. Additionally, the number of oligotrophic and copiotrophic bacteria was counted by the agar plate method. Results indicated a lack of site-specific distribution of bacterial abundance, in contrast to bacterial catabolic activity and community level physiological profiles. Community level physiological profiles revealed a common capacity of soil bacteria to intensively utilize polyols, which are cryoprotectants widely produced by Antarctic organisms, as well as site-specific phenolic compounds (vegetated habitats), amino acids/amines (moss habitats), carbohydrates and carboxylic acids (fell field habitat). It was concluded that the physiology of soil bacteria is habitat specific concerning both the rate of catabolic activity and pattern of carbon source utilization.     

Keratinophilic fungi in the antarctic environment

Results of a study on the diffusion of keratinophilic fungi in an Antarctic environment are given. Nine soil samples collected from as many sites along the coast of Ross Sea, and six dust samples inside the Italian scientific base were examined by direct inoculation and hair baiting methods for soil samples and plate dilution method for dust samples. As regards the variety of species isolated and the counting of the numbers of colonies, plate dilution method proved to be very effective.Aspergillus spp.,Cladosporium sp., Dematiaceae,Fusarium sp.,Geomyces pannorum v.pannorum, G. pannorum v.vinaceus, Mycelia sterilia,Penicillium spp. were isolated from soil. From the dust the following moulds were isolated:Aphanoascus fulvescens, Aspergillus sp.,Beauveria sp.,Chrysosporium carmichaelii, Dematiaceae, Geomyces pannorum v.pannorum, G. pannorum v.vinaceus, Malbranchea gypsea, Mycelia sterilia, Nectria inventa, Penicillium spp.,Scopulariopsis brevicaulis, Scopulariopsis sp. andTrichophyton mentagrophytes. The presence of four colonies ofTrichophyton mentagrophytes is emphasized and correlated with the anthropization process.     

Fungal community associated with marine macroalgae from Antarctica

Filamentous fungi and yeasts associated with the marine algae Adenocystis utricularis, Desmarestia anceps, and Palmaria decipiens from Antarctica were studied. A total of 75 fungal isolates, represented by 27 filamentous fungi and 48 yeasts, were isolated from the three algal species and identified by morphological, physiological, and sequence analyses of the internal transcribed spacer region and D1/D2 variable domains of the large-subunit rRNA gene. The filamentous fungi and yeasts obtained were identified as belonging to the genera Geomyces, Antarctomyces, Oidiodendron, Penicillium, Phaeosphaeria, Aureobasidium, Cryptococcus, Leucosporidium, Metschnikowia, and Rhodotorula. The prevalent species were the filamentous fungus Geomyces pannorum and the yeast Metschnikowia australis. Two fungal species isolated in our study, Antarctomyces psychrotrophicus and M. australis, are endemic to Antarctica. This work is the first study of fungi associated with Antarctic marine macroalgae, and contributes to the taxonomy and ecology of the marine fungi living in polar environments. These fungal species may have an important role in the ecosystem and in organic matter recycling.     

Halo- and psychrotolerant Geomyces fungi from arctic cryopegs and marine deposits

Comparative characterization of Geomyces isolates was performed. The isolates were obtained from Arctic cryopegs and the surrounding ancient marine deposits, from nonsaline permafrost soils, and from temperate environments. Microbiological (cultural and morphological) and molecular criteria were used to confirm the identification of the isolates as Geomyces pannorum. The isolates from cryopegs and surrounding marine deposits were shown to differ from those obtained from nonsaline soils and temperate environments in their ability to grow at negative temperatures (-2 degrees C) under increased salt concentration (10%). The results are discussed in relation to the possible inheritance of the adaptive characteristics acquired in specific environments.     

A research on the microfungal flora of somegreenhouse soils in the vicinity of LapsekiÇanakkale,Turkey

Qualitative and quantitative distributions of microfungal flora of the some greenhouse soils in the vicinity of Lapseki, Canakkale were studied. A total of 25 soil samples were collected from 10 cm depth in five greenhouses. Using soil dilution technique, 128 different microfungal isolates were obtained Isolates collected belonged to Oomycetes (1), Zygomycetes (7), Ascomycetes (9) and 96 belonging to the Deuteromycetes Fifteen isolates were classified as Mycelia Sterilia. The genera of microfungi most abundant in the greenhouse soils were Aspergillus, Penicillium, Geomyces, Exophiala and Fusarium. Qualitative and quantitative distributions of the Aspergillus were higher than those of the other genera. It was found that the maximum value was in greenhouse 5 and the minimum value was in greenhouse 4. The solarization method was the most effective in reducing fungal numbers.     

南极恩克斯堡岛土壤中可培养细菌多样性

【背景】南极地区环境苛刻植被稀少,无冰区面积约占其总面积的0.4%,但是土壤中存在着丰富的微生物群落,对于极地微生物资源仍需要进一步挖掘。【目的】获得南极恩克斯堡岛土壤中可培养细菌多样性信息。【方法】对来源于南极恩克斯堡岛的5个土壤样品采用直接涂布、微好氧富集和有氧富集后涂布3种方法进行细菌分离培养。【结果】共获得144株可培养细菌,分布于5门30个属。这些菌株来自变形菌门(Proteobacteria,38.9%)、放线菌门(Actinobacteria,34.0%)、厚壁菌门(Firmicutes,22.2%)、异常球菌-栖热菌门(Deinococcus-Thermus,3.5%)、拟杆菌门(Bacteroidetes,1.4%)等。不同土壤样品中可培养细菌多样性存在差异。企鹅粪土中Bacillus是主要属类;Pseudomonas、Streptomyces在植被覆盖区土壤中是优势菌属;Psychrobacter在湖水边土壤中是优势属类,Flavobacterium、Chryseobacterium仅从该样品中分离得到;放线菌门类在干燥土壤样中占优势,Pseudarthrobacter、Rhodococcus、Microbacterium等属仅从这类干燥样品中分离出。经16S rRNA基因序列对比发现,有2株菌为潜在新种。【结论】南极恩克斯堡岛土壤中存在可开发利用的菌种资源,本文为研究该地细菌多样性提供一定的基础数据。     

Diversity and bioprospecting of fungal communities associated with endemic and cold-adapted macroalgae in Antarctica

We surveyed the distribution and diversity of fungi associated with eight macroalgae from Antarctica and their capability to produce bioactive compounds. The collections yielded 148 fungal isolates, which were identified using molecular methods as belonging to 21 genera and 50 taxa. The most frequent taxa were Geomyces species (sp.), Penicillium sp. and Metschnikowia australis. Seven fungal isolates associated with the endemic Antarctic macroalgae Monostroma hariotii (Chlorophyte) displayed high internal transcribed spacer sequences similarities with the psychrophilic pathogenic fungus Geomyces destructans. Thirty-three fungal singletons (66%) were identified, representing rare components of the fungal communities. The fungal communities displayed high diversity, richness and dominance indices; however, rarefaction curves indicated that not all of the fungal diversity present was recovered. Penicillium sp. UFMGCB 6034 and Penicillium sp. UFMGCB 6120, recovered from the endemic species Palmaria decipiens (Rhodophyte) and M. hariotii, respectively, yielded extracts with high and selective antifungal and/or trypanocidal activities, in which a preliminary spectral analysis using proton nuclear magnetic resonance spectroscopy indicated the presence of highly functionalised aromatic compounds. These results suggest that the endemic and cold-adapted macroalgae of Antarctica shelter a rich, diversity and complex fungal communities consisting of a few dominant indigenous or mesophilic cold-adapted species, and a large number of rare and/or endemic taxa, which may provide an interesting model of algal–fungal interactions under extreme conditions as well as a potential source of bioactive compounds.     

Bacterial diversity of a soil sample from Schirmacher Oasis, Antarctica.

The bacterial diversity of a soil sample collected in the vicinity of Lake Zub, Schirmacher Oasis, Antarctica, was determined both by establishing pure colonies of culturable bacteria and by cloning the total 16S rDNA of the soil and establishing the phylogeny of the clones. Analysis of the 16S rRNA gene clones indicated that the bacteria belonged to the classes alpha-proteobacteria, beta-proteobacteria, gamma-proteobacteria, Gemmatimonas, Bacteriodetes, Actinobacteria, Chloroflexi and Chlamydiae. In addition, seven clones were categorized as unidentified and unculturable in the classes of beta-Proteobacteria, Actinobacteria, Chloroflexi and Chlamydiae. Further, the culturable bacteria from the same site were identified as belonging to the genera Pseudomonas, Sphingobacterium, Arthrobacter, Micrococcus, Brevondimonas, Rhodococcus and Microbacterium. These results identify for the first time the presence of bacteria belonging to the genera Brevundimonas, Microbacterium, Rhodococcus, Serratia, Enterobacter, Rhodopseudomonas, Sphingomonas, Acidovorax, Burkholderia, Nevskia, Gemmatimonas, Xanthomonas and Flexibacter in Antarctica. Further, comparison of the Antarctic soil bacterial diversity with other cold habitats of Antarctica like from sediments, ice and cyanobacterial mat samples indicated that the bacterial diversity in soil was similar to the diversity observed in the continental shelf sediment sample. The Antarctic soil clones also resembled the bacterial diversity of soils from other geographical regions, but were unique in that none of the clones from the soil belonged to the uncultured Y, O, G, A and B groups common to all soil samples.