Isolation of keratinolytic fungi from a coal mine dump

During the study of fungal succesion in the coal mine dump in Brzezinka (Poland), soil samples were examined for keratinolytic fungi. These micro-organisms were rather poorly represented in the area studied. Out of 300 soil samples examined, only 48 (16%) were positive for keratinolytic fungi.Trichophyton ajelloi andArthroderma curreyi were the prevailing species. These species occurred practically at two locations, i.e. on the naked carbon rocks inhabited by algae crops (chiefly byCyanophyta) and in the pine litter. It can be supposed that the occurrence of keratinolytic fungi was more dependent on the favourable general conditions such as increasing organic matter content, microflora, and humidity than on the presence of keratin remains in the soil. Because of the lack of potentially pathogenic fungi, the coal mine dump examined cannot be considered as an important source of fungal infection.     

New lipid‐producing,cold‐tolerant yellow‐green alga isolated from the rocky mountains of colorado

A new strain of yellow‐green algae (Xanthophyceae, Heterokonta), tentatively named Heterococcus sp. DN1 (UTEX accession number UTEX ZZ885), was discovered among snow fields in the Rocky Mountains. Axenic cultures of H. sp. DN1 were isolated and their cellular morphology, growth, and composition of lipids were characterized. H. sp. DN1 was found to grow at temperatures approaching freezing to accumulate large intracellular stores of lipids. H. sp. DN1 produces the highest quantity of lipids when grown undisturbed with high light in low temperatures_. Of particular interest was the accumulation of eicosapentaenoic acid, known to be important for human nutrition, and palmitoleic acid, known to improve biodiesel feedstock properties. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:853–861, 2013     

Identification and characterization of a novel salt‐tolerant esterase from a Tibetan glacier metagenomic library

A salt‐tolerant esterase, designated H9Est, was identified from a metagenomic library of the Karuola glacier. H9Est gene comprised 1071 bp and encoded a polypeptide of 357 amino acids with a molecular mass of 40 kDa. Sequence analysis revealed that H9Est belonged to the family IV of bacterial lypolitic enzyme. H9Est was overexpressed in Escherichia coli and the purified enzyme showed hydrolytic activity towards p‐nitrophenyl esters with carbon chain from 2 to 8. The optimal esterase activity was at 40°C and pH 8.0 and the enzyme retained its activity towards some miscible organic solvents such as polyethylene glycol. A three‐dimensional model of H9Est revealed that S200, D294, and H324 formed the H9Est catalytic triad. Circular Dichroism spectra and molecular dynamic simulation indicated that the esterase had a wide denaturation temperature range and flexible loops that would be beneficial for H9Est performance at low temperatures while retaining heat‐resistant features. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:890–899, 2015     

Isolation of fungi from soil using the keratin-baiting technique

Of 236 soil samples baited with wool, some 71.2% (168) produced fungal growth. Gliocladium (25.0%), Paecilomyces (14.8%), Trichophyton (11.9%) species were the most prevalent in soil moistened with sterile distilled water (SDW) containing antibiotics. On the other hand, in soil moistened with SDW only, Trichophyton (32.6%), Paecilomyces (27.5%), Diheterospora (16.5%), Gliocladium (13.6%) and Fusarium (13.1%) species were more common. Of the known potential pathogens, the Paecilomyces and Fusarium species were frequently isolated from soil collected in parks, cleared areas, paddocks, rivers and roadsides. All five sites are areas of human and animal activity. The keratinolytic species were Microsporum cookei, M. gypseum complex, Trichophyton ajelloi and T. terrestre, which are regarded as nonpathogens with the exception of M. cookei and M. gypseum complex.     

自然演替过程中铜尾矿土壤酶活性的变化

铜陵矿区堆放了大量不同时期的铜尾矿,形成大量铜尾矿废弃地,其中的大部分铜尾矿废弃地处于自然生态恢复状态。并形成典型的原生演替系列。为探讨废弃地自然生态恢复中不同演替阶段植物群落变化与土壤酶活性变化的关系,对铜陵3处不同时期弃置的尾矿废弃地进行了调查。随着自然生态系统演替的进行,尾矿中土壤酶活性逐步提高,维管植物群落下尾矿中土壤酶活性〉隐花植物结皮下尾矿中土壤酶活性〉裸地处土壤酶活性;位于植物群落下的尾矿废弃地中脲酶、碱性磷酸酶、过氧化氢酶的活性从表层至下层逐渐降低。不同植物群落对尾矿中土壤酶活性影响有很大差异,位于木贼群落下的尾矿中几种土壤酶的活性高于位于白茅群落和中华结缕草群落下的尾矿中酶的活性。相关分析表明,土壤酶活性与有机质、总N表现出显著的正相关。     

Characterization of an ethanol‐tolerant 1,4‐β‐xylosidase produced by Pichia membranifaciens

Aims: The purification and biochemical properties of the 1,4‐β‐xylosidase of an oenological yeast were investigated. Methods and Results: An ethanol‐tolerant 1,4‐β‐xylosidase was purified from cultures of a strain of Pichia membranifaciens grown on xylan at 28°C. The enzyme was purified by sequential chromatography on DEAE cellulose and Sephadex G‐100. The relative molecular mass of the enzyme was determined to be 50 kDa by SDS‐PAGE. The activity of 1,4‐β‐xylosidase was optimum at pH 6·0 and at 35°C. The activity had a K_m of 0·48 ± 0·06 mmol l^?1 and a V_max of 7·4 ± 0·1 μmol min^?1 mg^?1 protein for p‐nitrophenyl‐β‐d ‐xylopyranoside. Conclusions: The enzyme characteristics (pH and thermal stability, low inhibition rate by glucose and ethanol tolerance) make this enzyme a good candidate to be used in enzymatic production of xylose and improvement of hemicellulose saccharification for production of bioethanol. Significance and Impact of the Study: This study may be useful for assessing the ability of the 1,4‐β‐xylosidase from P. membranifaciens to be used in the bioethanol production process.     

湖北铜绿山古铜矿遗址区木本植物对重金属富集能力的分析

对湖北铜绿山古铜矿遗址区的木本植物种类组成及生长概况进行了调查,并采用原子吸收光谱法测定了木本植物地上部和地下部及其根际土壤中重金属含量;在此基础上,分析了各树种对Cu、Cd和Pb的富集作用,并对木本植物的生态修复意义进行了初步评价。结果表明:在该古铜矿遗址区共有木本植物13科14属14种,其中乔木13种、灌木1种;木本植物的树龄4~25 a、株高3.7~22.6 m、胸径5.3~29.0 cm,长势均较好。木本植物根际土壤中Cu、Cd、Pb、Cr、Mn和Zn的平均含量分别为3 166.73、3.66、137.06、31.32、1 774.43和208.32 mg·kg-1,其中,土壤中Cu、Cd和Pb的含量达到重度污染等级。各树种地上部和地下部的Cu、Cd和Pb的含量差异明显,且均为Cu含量最高、Cd含量最低;各树种的地上部和地下部对Cu、Cd和Pb的富集系数及综合富集系数也均有明显差异,但对Cd的富集系数均最大。法国冬青〔Viburnum odoratissimum var.awabuki(K.Koch)Zabel ex Rumpl.〕、梧桐〔Firmiana platanifolia(Linn.f.)Marsili〕、桂花〔Osmanthus fragrans(Thunb.)Lour.〕和刺槐(Robinia pseudoacacia Linn.)地上部的综合富集系数较高,苦楝(Melia azedarach Linn.)、女贞(Ligustrum lucidum Ait.)、法国冬青和樟树〔Cinnamomum camphora(Linn.)Presl〕地下部的综合富集系数较高;总体上看,法国冬青、苦楝、女贞、梧桐和桂花对Cu、Cd和Pb的平均综合富集系数均大于1,具有较高的重金属富集能力。根据研究结果,建议在Pb污染区域可选择栽植二球悬铃木〔Platanus×acerifolia(Ait.)Willd.〕和构树〔Broussonetia papyrifera(Linn.)L’Hert.ex Vent.〕等树种,在Cd污染区域可选择栽植法国冬青、梧桐、刺槐和苦楝等树种,而在Cu-Cd-Pb复合污染区域应栽植法国冬青、苦楝、女贞、梧桐、桂花和刺槐等树种。     

狮子山铜尾矿植物对铜的吸收及土壤特的影响

我国铜矿储藏丰富,铜矿开采带来巨大经济利益的同时,也对生态环境造成极大的破坏,这种恶劣的环境严重阻碍了植物的定居,但是自然界物种繁多,总有一些植物能适应这种环境而生存下来.本文通过对狮子山优势植物吸收和积累铜的分析,发现这些植物均能富集较多的铜,在土壤铜含量很高的情况下,依然生长旺盛,没有出现受害症状,成为尾矿上的优势种,并形成了单优群落或多优小群落.这些植物的存在改变了土壤的理化特性,降低了土壤中的重金属的含量,提高了土壤的全N、全P、全K和有机质含量,一定程度上改善了土壤的不良环境,在尾矿的植被恢复和土壤修复中起着非常重要的作用.     

Genotype‐by‐sequencing of the plant‐pathogenic fungi Pyrenophora teres and Sphaerulina musiva utilizing Ion Torrent sequence technology

Genetic and genomics tools to characterize host–pathogen interactions are disproportionately directed to the host because of the focus on resistance. However, understanding the genetics of pathogen virulence is equally important and has been limited by the high cost of de novo genotyping of species with limited marker data. Non‐resource‐prohibitive methods that overcome the limitation of genotyping are now available through genotype‐by‐sequencing (GBS). The use of a two‐enzyme restriction‐associated DNA (RAD)‐GBS method adapted for Ion Torrent sequencing technology provided robust and reproducible high‐density genotyping of several fungal species. A total of 5783 and 2373 unique loci, ‘sequence tags’, containing 16 441 and 9992 single nucleotide polymorphisms (SNPs) were identified and characterized from natural populations of Pyrenophora teres f. maculata and Sphaerulina musiva, respectively. The data generated from the P. teres f. maculata natural population were used in association mapping analysis to map the mating‐type gene to high resolution. To further validate the methodology, a biparental population of P. teres f. teres, previously used to develop a genetic map utilizing simple sequence repeat (SSR) and amplified fragment length polymorphism (AFLP) markers, was re‐analysed using the SNP markers generated from this protocol. A robust genetic map containing 1393 SNPs on 997 sequence tags spread across 15 linkage groups with anchored reference markers was generated from the P. teres f. teres biparental population. The robust high‐density markers generated using this protocol will allow positional cloning in biparental fungal populations, association mapping of natural fungal populations and population genetics studies.     

Effects of small increases in copper levels on culturable basidiomycetous yeasts in low‐nutrient soils

Aims: Investigating the effect of perturbations, with relatively low Cu concentrations, on yeast community composition in low‐nutrient virgin soil. Methods and Results: Culturable soil yeast populations were monitored at an experimental site treated with the fungicide copper oxychloride (10 mg Cu per kg soil). Yeast numbers were unaffected by additional Cu; however, a shift in yeast community composition from Hymenomycetes to Urediniomycetes species occurred. Subsequent growth experiments conducted with a synthetic liquid medium revealed that hymenomycetous and urediniomycetous yeasts were affected differently by 1 and 10 mg l^?1 Cu. Soil microcosm experiments then indicated that additional 10 mg kg^?1 Cu may improve the competitive ability of urediniomycetous yeasts in the presence of hymenomycetous yeasts. Conclusions: The shift from hymenomycetous to urediniomycetous yeasts, as a result of slightly increased soil Cu levels, may be because of hymenomycetous yeasts being more sensitive to elevated Cu levels and urediniomycetous yeasts having an improved competitive ability in the presence of elevated Cu levels. Significance and Impact of the Study: Yeast community composition of pristine low‐nutrient soils may change as a result of perturbations with relatively low concentrations of Cu. Urediniomycetous yeasts should be studied as potential bio‐indicators of Cu perturbations.     

Isolation and partial purification of fungal ligninolytic enzymes from the forest soil fungi isolated from Bhadra Wildlife Sanctuary

Screening was done for the isolation of effective lignin degraders from the forest soil samples, by providing lignin as a carbon source through the enrichment method, which leads to the isolation of 8 effective fungal isolates among 14 isolates. Submerged fermentation was done for the production of ligninolytic enzymes with the effective microorganisms by providing Guiaicol as a carbon source. The assay of laccase, lignin peroxidise activity and specific activity was done after the incubation intervals of 2, 4, 6, 7, 8, 10 and 12 days at 27±2℃ under shake culture condition. Partially purified protein content was estimated by using Lowry＇s method. Pleurotus sp. and Phanerochaetae chrysosporium are more effective at the 2nd and 7th days of incubation for the production of laccase and lignin peroxidases among the effective isolates.     

Isolation of opportunistic fungi from bronchoalveolar lavage of compromised hosts in Isfahan,Iran

In this study, bronchoalveolar lavage (BAL) specimens from 247 immunocompromised patients were investigated for the incidence of opportunistic fungi. In the direct examination and culture of the specimens, 5 (2.02%) of filamentous fungi and 55 (22.26%) yeasts were isolated and identified as follows: Aspergillus fumigatus (2), A. terreus (1), A. nidulans (1), Mucorsp. (1), Candida albicans (29), C. glabrata (3), C. Parapsilosis (1), Trichosporon beigelii (1), Candida sp. (13) and unknown yeasts (8). In addition, one strain of Nocardia asteroides and two strains of Streptomyces sp. were isolated.     

Intensification of biosorption of copper ions from solution by the yeast Saccharomyces cerevisiae in magnetic field

We studied the possibility of intensification of biosorption of copper ions by the yeast Saccharomyces cerevisiae 1968 from a copper sulfate solution by immersing a metal 80-rod headpiece into the solution and applying an external magnetic field. The field was parallel or perpendicular to the axes of headpiece rods. It was shown that intensification of extraction of copper ions at various geometries of the system differed insignificantly and that copper ions were extracted from the solution via biosoprtion and cementation at the metal headpiece.     

Copper‐resistant halophilic bacterium isolated from the polluted Maruit Lake,Egypt

Aims: To isolate and characterize copper‐resistant halophilic bacteria from the polluted Maruit Lake, Egypt and identify the role of plasmids in toxic metal resistance. Methods and Results: We isolated strain MA2, showing high copper resistance up to the 1·5 mmol l^?1 concentration; it was also resistant to other metals such as nickel, cobalt and zinc and a group of antibiotics. Partial 16S rRNA analysis revealed that strain MA2 belonged to the genus Halomonas. Copper uptake, measured by atomic absorption spectrophotometery, was higher in the absence of NaCl than in the presence of 0·5–1·0 mol l^?1 NaCl during 5–15 min of incubation. Cell fractionation and electron microscopic observation clarified that most of the copper accumulated in the outer membrane and periplasmic fractions of the cells. Plasmid screening yielded two plasmids: pMA21 (11 kb) and pMA22 (5 kb). Plasmid curing resulted in a strain that lost both the plasmids and was sensitive to cobalt and chromate but not copper, nickel and zinc. This cured strain also showed weak growth in the presence of 0·5–1·0 mol l^?1 NaCl. Partial sequencing of both plasmids led to the identification of different toxic metals transporters but copper transporters were not identified. Conclusions: The highest cell viability was found in the presence of 1·0 mol l^?1 NaCl at different copper concentrations, and copper uptake was optimal in the absence of NaCl. Plasmid pMA21 encoded chromate, cobalt, zinc and cadmium transporters, whereas pMA22 encoded specific zinc and RND (resistance, nodulation, cell division) efflux transporters as well as different kinds of metabolic enzymes. Copper resistance was mainly incorporated in the chromosome. Significance and Impact of the Study: Strain MA2 is a fast and efficient tool for copper bioremediation and the isolated plasmids show significant characteristics of both toxic metal and antibiotic resistance.     

Effects of mine drainage on the river hayle,cornwall a) factors affecting concentrations of copper,zinc and iron in water,sediments and dominant invertebrate fauna

Concentrations of copper, zinc and iron were measured in waters, sediments and invertebrates collected from the River Hayle. In river water at least 70% of copper and iron was associated with the particulate fraction whereas 80% of zinc was in the soluble form. Although total concentrations of zinc in water exceeded those of copper approximately ten fold, copper predominated over zinc in the sediments by a factor of approximately three. Iron was the most abundant metal recorded in both water and sediments.Seasonal differences in total metal content of waters suggested that concentrations of copper, zinc and iron increased during. periods of high flow and decreased during lower flows. Copper concentrations in the sediment, unlike zinc and iron, showed markedly higher values during the summer sampling period when flows were minimal.In the free-living Trichoptera larvae, concentrations of copper and zinc in the tissue appeared to follow copper and zinc levels in the water. Similar relationships in Odonata and Plecoptera larvae were not obtained. Factors affecting animal/metal relationships are discussed with particular reference to adaptation shown by organisms exposed to high concentrations of heavy metals in their environment.Department of Applied Biology, Cambridge     

Phylogenetic analysis of manganese-oxidizing fungi isolated from manganese-rich aquatic environments in Hokkaido, Japan

Three strains of Mn-oxidizing fungi were isolated from manganese-rich aquatic environments: sediment in a stream (Komanoyu) in Mori-machi and inflow to an artificial wetland in Kaminokuni-cho, Hokkaido, Japan. The characteristics of each strain were then established. Genetic analysis based on the ribosomal RNA (rRNA) gene was performed to clarify their classification. The sequences of the 18S rRNA and internal transcribed spacer (ITS1)-5.8S rRNA-ITS2 genes showed that all three strains are Ascomycetes. Based on its morphology, it seems probable that the KY-1 strain from Mori-machi belongs to the genus Phoma or Ampelomyces. The phylogenetic analysis indicates that this strain belongs to Phoma rather than Ampelomyces. Morphological identification of WL-1 and WL-2 strains from Kaminokuni-cho was impossible because of the lack of a sexual stage and specific organs. Phylogenetic analysis of the sequence in the ITS1-5.8S rRNA-ITS2 gene suggests that the WL-1 strain corresponds to Paraconyothyrium sporulosum and that WL-2 also belongs to the genus Paraconiothyrium. Because the ability to oxidize Mn has not been evaluated for most species of Phoma or Paraconiothyrium (Coniothyrium), further study is needed to confirm the status of these three strains.