后基因组时代的真菌天然产物发现

真菌产生的次级代谢产物是新药发现的重要来源之一,然而近年来传统的真菌天然产物发现方法在大量真菌基因组测序完成的时代遇到了很大的挑战。如何利用这些基因组数据来发现真菌中新的天然产物已成为后基因组时代天然产物发现的研究重点和热点。本综述先后介绍了真菌天然产物的类型及其相应基因簇和骨架酶的特征,基于基因组挖掘技术发展而来的天然产物发现新策略,以及利用合成生物学理念和技术在真菌天然产物发现中的应用现状,最后展望了后基因组时代中的天然产物发现研究前沿及基因组数据在后基因组时代对真菌天然产物发现的应用前景。     

Marine microorganisms as biocontrol agents against fungal phytopathogens and mycotoxins

Mycotoxins are a serious food safety concern for human and animal health. Much attention should be paid to the dietary exposure to mycotoxins in order to minimise the risk of mycotoxin contamination in the food chain. Among the reported strategies to manage the mycotoxin contamination into food and feed, biological control seems a promising approach, depending on their biological origins, and on the use of living organisms or their derivatives. Marine microorganisms have developed unique metabolic and physiological capabilities to thrive in extreme habitats and produce novel metabolites which are not often present in microbes of terrestrial origin. Some marine bacteria and fungi have a good potential for the control of fungal phytopathogens and mycotoxins. Biologists and chemists are needed to work together to explore the storehouse of marine microorganisms and marine active metabolites, because marine bacteria and fungi have a huge potential for practical application in biocontrol of fungal phytopathogens and preventing mycotoxin contamination.     

Evolving challenges and strategies for fungal control in the food supply chain

Fungi that spoil foods or infect crops can have major socioeconomic impacts, posing threats to food security. The strategies needed to manage these fungi are evolving, given the growing incidence of fungicide resistance, tightening regulations of chemicals use and market trends imposing new food-preservation challenges. For example, alternative methods for crop protection such as RNA-based fungicides, biocontrol, or stimulation of natural plant defences may lessen concerns like environmental toxicity of chemical fungicides. There is renewed focus on natural product preservatives and fungicides, which can bypass regulations for ‘clean label’ food products. These require investment to find effective, safe activities within complex mixtures such as plant extracts. Alternatively, physical measures may be one key for fungal control, such as polymer materials which passively resist attachment and colonization by fungi. Reducing or replacing traditional chlorine treatments (e.g. of post-harvest produce) is desirable to limit formation of disinfection by-products. In addition, the current growth in lower sugar food products can alter metabolic routing of carbon utilization in spoilage yeasts, with implications for efficacy of food preservatives acting via metabolism. The use of preservative or fungicide combinations, while involving more than one chemical, can reduce total chemicals usage where these act synergistically. Such approaches might also help target different subpopulations within heteroresistant fungal populations. These approaches are discussed in the context of current challenges for food preservation, focussing on pre-harvest fungal control, fresh produce and stored food preservation. Several strategies show growing potential for mitigating or reversing the risks posed by fungi in the food supply chain.     

Perspectives on the potential of entomopathogenic fungi in biological control of ticks

Ticks are serious health threats for humans, and both domestic and wild animals. Ticks are controlled mostly by application of chemical products; but these acaricides have several negative side effects, including toxicity to animals, environmental contamination, and induction of chemical resistance in some tick populations. Entomopathogenic fungi infect arthropods in nature and can occur at enzootic or epizootic levels in their host populations. Laboratory studies clearly demonstrate that these fungi can cause high mortality in all developmental stages of several tick species, and also reduce oviposition of infected engorged females. Tick mortality following application of fungi in the field, however, often is less than that suggested by laboratory tests. This is due to many negative biotic and climatic factors. To increase efficacy of fungal agents for biological control of ticks under natural conditions, several points need consideration: (1) select effective isolates (viz., high virulence; and tolerance to high temperature, ultraviolet radiation and desiccation); (2) understand the main factors that affect virulence of fungal isolates to their target arthropods including the role of toxic metabolites of the fungal isolates; and (3) define with more precision the immune response of ticks to infection by entomopathogenic fungi. The current study reviews recent literature on biological control of ticks, and comments on the relevance of these results to advancing the development of fungal biocontrol agents, including improving formulation of fungal spores for use in tick control, and using entomopathogenic fungi in integrated pest (tick) management programs.     

农田土壤中铜的主要输入途径及其污染风险控制

综述农田土壤中畜禽粪便、杀菌剂、再生水和化肥等途径对铜的输入及其污染风险,以土壤理化性质为切入点概述外源铜进入土壤后的吸附-解吸和移动性特点,探讨不同作物类型和土地利用方式对农产品中铜含量和对通过膳食途径铜健康风险的影响,并针对实际生产提出规避农田土壤铜污染风险的有效措施.     

Assessment of mycoflora and infestation of insects,vector of <Emphasis Type="Italic">Aspergillus</Emphasis> section <Emphasis Type="Italic">Flavi</Emphasis>, in stored peanut from Argentina

The occurrence of spoilage fungi and Aspergillus section Flavi populations, the aflatoxins incidence, the role of insects as vectors of mycotoxin-producing fungi and the AFs-producing ability of the isolated species throughout the peanut (Arachis hypogaea L.) storage period were evaluated. Analyses of fungal populations from 95 peanut seed samples did not demonstrate significant differences between the incidences in each sampling period. Aspergillus section Flavi were isolated during all incubation periods. Cryptolestes spp. (Coleoptera: Cucujidae) were collected in August, September and October with 18, 16 and 28% of peanut samples contaminated, respectively. Insects isolated during August showed 69% of Aspergillus section Flavi contamination. A. flavus was the most frequently isolated (79%) from peanut seeds and from insect (59%). The greater levels of AFB₁ were detected in September and October with a mean of 68.86 μg/kg and 69.12 μg/kg respectively. The highest proportion of A. flavus toxigenic strains (87.5%) was obtained in June. The presence of Aspergillus section Flavi and insect vectors of aflatoxigenic fungi presented a potential risk for aflatoxin production during the peanut storage period. Integrated management of fungi and insect vectors is in progress.     

STUDIES ON SEED HEALTH

An account is given of the survey made in 1940-55 to determine the incidence of the seed-borne fungal parasites of flax. It was shown that fibre flax seed produced in eastern and south-eastern England was relatively free from contamination with Colletotrichum linicola, Polyspora lini and Phoma sp. and that there was no build-up of infection in seed stocks in these areas. In the wetter and cooler regions west and north of the Pennines, conditions were more favourable for these fungi, and seed contamination tended to increase.
Seed contamination with Botrytis cinerea was general throughout the United Kingdom, with a tendency to be higher in coastal and upland areas. The incidence of Fusarium lini and Alternaria linicola in fibre flax seed produced in the United Kingdom was negligible.
The examination of seed samples from other countries indicated the possible importance of P. lini in Denmark, of B. cinerea and C. linicola in the Netherlands, of A. linicola as a contaminant of oil flax seed in Sweden, and of C. linicola in parts of Canada.
Seed disinfection, using a thiram product, proved useful for the control of flax diseases in general, but should be used as an ancillary to the method of avoiding seed infection by limiting the production of seed flax to areas where the risk of contamination is slight or absent. Where there is any risk of a build-up of contamination, seed disinfection should be carried out as a routine practice each season.     

食药用真菌多糖抗氧化作用研究进展

食药用真菌多糖由于其独特的生理活性,目前正成为国内外众多学科领域研究的热点之一。综述了食药用真菌多糖抗氧化作用研究现状、作用机理及其构效与量效关系,并对其发展前景进行了展望,旨在为其深入研究和开发利用提供参考。     

Entomopathogenic fungi against South American tick species

Ticks are parasites of serious concern for humans, domesticated animals and wild animals. Despite scientific advances, in South America the principal control method for ticks is the use of chemical acaricides. Indiscriminate use of these products causes environmental pollution, food contamination and development of tick resistance to acaricides. In vitro studies and field trials have demonstrated that pathogenic fungal isolates not only cause mortality of many tick species, but also reduce subsequent generations due to effects on their reproductive efficacy. Accordingly, this review presents results of several studies which were conducted in South America. Furthermore, it outlines current information on fungal pathogens of ticks and discusses the need to develop and implement effective strategies for use of entomopathogenic fungi to control ticks in the near future.     

Case Report: A Case of Chromoblastomycosis Caused by Fonsecaea pedrosoi in Vietnam

Mycopathologia - Chromoblastomycosis is a chronic fungal infection of the skin and subcutaneous tissues caused by different melanized fungi. The disease occurs worldwide, particularly in tropical...     

Protection of melanized Cryptococcus neoformans from lethal dose gamma irradiation involves changes in melanin's chemical structure and paramagnetism

Certain fungi thrive in highly radioactive environments including the defunct Chernobyl nuclear reactor. Cryptococcus neoformans (C. neoformans), which uses L-3,4-dihydroxyphenylalanine (L-DOPA) to produce melanin, was used here to investigate how gamma radiation under aqueous aerobic conditions affects the properties of melanin, with the aim of gaining insight into its radioprotective role. Exposure of melanized fungal cell in aqueous suspensions to doses of γ-radiation capable of killing 50 to 80% of the cells did not lead to a detectable loss of melanin integrity according to EPR spectra of melanin radicals. Moreover, upon UV-visible (Xe-lamp) illumination of melanized cells, the increase in radical population was unchanged after γ-irradiation. Gamma-irradiation of frozen cell suspensions and storage of samples for several days at 77 K however, produced melanin modification noted by a reduced radical population and reduced photoresponse. More direct evidence for structural modification of melanin came from the detection of soluble products with absorbance maxima near 260 nm in supernatants collected after γ-irradiation of cells and cell-free melanin. These products, which include thiobarbituric acid (TBA)-reactive aldehydes, were also generated by Fenton reagent treatment of cells and cell-free melanin. In an assay of melanin integrity based on the metal (Bi(+3)) binding capacity of cells, no detectable loss in binding was detected after γ-irradiation. Our results show that melanin in C. neoformans cells is susceptible to some damage by hydroxyl radical formed in lethal radioactive aqueous environments and serves a protective role in melanized fungi that involves sacrificial breakdown.     

Microbiological control in stem cell banks: approaches to standardisation

The transplant of cells of human origin is an increasingly complex sector of medicine which entails great opportunities for the treatment of a range of diseases. Stem cell banks should assure the quality, traceability and safety of cultures for transplantation and must implement an effective programme to prevent contamination of the final product. In donors, the presence of infectious micro-organisms, like human immunodeficiency virus, hepatitis B virus, hepatitis C virus and human T cell lymphotrophic virus, should be evaluated in addition to the possibility of other new infectious agents (e.g. transmissible spongiform encephalopathies and severe acute respiratory syndrome). The introduction of the nucleic acid amplification can avoid the window period of these viral infections. Contamination from the laboratory environment can be achieved by routine screening for bacteria, fungi, yeast and mycoplasma by European pharmacopoeia tests. Fastidious micro-organisms, and an adventitious or endogenous virus, is a well-known fact that will also have to be considered for processes involving in vitro culture of stem cells. It is also a standard part of current good practice in stem cell banks to carry out routine environmental microbiological monitoring of the cleanrooms where the cell cultures and their products are prepared. The risk of viral contamination from products of animal origin, like bovine serum and mouse fibroblasts as a “feeder layer” for the development of embryonic cell lines, should also be considered. Stem cell lines should be tested for prion particles and a virus of animal origin that assure an acceptable quality.     

Determining Fungi rRNA Copy Number by PCR

The goal of this project is to improve the quantification of indoor fungal pollutants via the specific application of quantitative PCR (qPCR). Improvement will be made in the controls used in current qPCR applications. This work focuses on the use of two separate controls within a standard qPCR reaction. The first control developed was the internal standard control gene, benA. This gene encodes for β-tubulin and was selected based on its single-copy nature. The second control developed was the standard control plasmid, which contained a fragment of the ribosomal RNA (rRNA) gene and produced a specific PCR product. The results confirm the multicopy nature of the rRNA region in several filamentous fungi and show that we can quantify fungi of unknown genome size over a range of spore extractions by inclusion of these two standard controls. Advances in qPCR have led to extremely sensitive and quantitative methods for single-copy genes; however, it has not been well established that the rRNA can be used to quantitate fungal contamination. We report on the use of qPCR, combined with two controls, to identify and quantify indoor fungal contaminants with a greater degree of confidence than has been achieved previously. Advances in indoor environmental health have demonstrated that contamination of the built environment by the filamentous fungi has adverse impacts on the health of building occupants. This study meets the need for more accurate and reliable methods for fungal identification and quantitation in the indoor environment.     

Competing Risks and Time‐Dependent Covariates

Time‐dependent covariates are frequently encountered in regression analysis for event history data and competing risks. They are often essential predictors, which cannot be substituted by time‐fixed covariates. This study briefly recalls the different types of time‐dependent covariates, as classified by Kalbfleisch and Prentice [The Statistical Analysis of Failure Time Data, Wiley, New York, 2002] with the intent of clarifying their role and emphasizing the limitations in standard survival models and in the competing risks setting. If random (internal) time‐dependent covariates are to be included in the modeling process, then it is still possible to estimate cause‐specific hazards but prediction of the cumulative incidences and survival probabilities based on these is no longer feasible. This article aims at providing some possible strategies for dealing with these prediction problems. In a multi‐state framework, a first approach uses internal covariates to define additional (intermediate) transient states in the competing risks model. Another approach is to apply the landmark analysis as described by van Houwelingen [Scandinavian Journal of Statistics 2007, 34 , 70–85] in order to study cumulative incidences at different subintervals of the entire study period. The final strategy is to extend the competing risks model by considering all the possible combinations between internal covariate levels and cause‐specific events as final states. In all of those proposals, it is possible to estimate the changes/differences of the cumulative risks associated with simple internal covariates. An illustrative example based on bone marrow transplant data is presented in order to compare the different methods.     

Quality Control of Fungal and Viral Biocontrol Agents - Assurance of Product Performance

An essential feature of the production of all microbial control agents is an effective quality control system. Well-defined product specifications with accompanying quality control procedures help to maximize product performance, ensure product safety, standardize manufacturing costs and reduce the risks of supply failure, thus building user confidence. A production system that does not have a quality control system is one whose output is uncontrolled and a lack of thorough quality feedback can result in batches of product with variable concentrations of active agent. This results in products with variable performance leading to control failures by users and serious loss of user confidence. Strict quality control procedures are not only essential for product consistency, but also for safety. Where quality control is inadequate, microbial contamination of the final product is inevitable. In most of such cases this will merely lead to a loss of efficacy due to dilution of the active ingredient by competing microorganisms, but also the potential of producing human pathogens must be ruled out. Recognition of contaminants and quantification of the degree of contamination are therefore important in determining any possible risk to human health. Many low technology production systems in use around the world have minimal or no quality control procedures. This is unacceptable and can damage the reputation of microbial control in addition to possibly posing health risks to those that produce or are exposed to the product. Two case studies from developing countries, are used to illustrate how the lack of quality control procedures can lead to the production of low viability, highly contaminated products with low or negligible concentrations of the active ingredient. However, it is also demonstrated that low technology production systems in developing countries can produce high quality products, provided appropriate quality control procedures are firmly implemented. It must be recognized that quality control procedures can be more complex and technologically demanding than the production procedures themselves, but it is largely on the effectiveness of these control procedures that the long-term acceptability of fungal and viral products depends. This paper details the quality control procedures considered necessary in the mass production of fungi and viruses for use as biocontrol agents, and attempts to suggest reasonable standards that can be achieved by all producers.     

Fungi in bottled water: a case study of a production plant

A one-year fungal survey of a water bottling plant was conducted in order to evaluate the incidence and fluctuations of the mycobiota. The dominant fungal genera in order of highest numbers isolated were Penicillium, Cladosporium and Trichoderma followed by Aspergillus, Paecilomyces, and others. As expected, the highest number of isolates were collected during the warmer months, particularly May and June. Indeed during these two months there were more fungi present in the water, indicating that during those times of the year when fungal contamination is high, 0.4 mm filters should be changed on a more regular basis. In order to assess whether contamination was single or multi-loci, molecular methods based on the PCR were used for Penicillium brevicompactum. Overall, fungal contamination arose from multiple sources. Some P. brevicompactum strains were very "alike" and were detected during different sampling times, indicating that they were endemic to the plant. There was no evidence to suggest that fungi detected in the source water passed through to other parts of the plant. However, there was evidence that fungal strains isolated from the water filter were detected elsewhere in the factory, confirming the need to change filters more regularly during periods of high fungal contamination. In order to improve quality control a HACCP programme was implemented and Best Practice Guidelines introduced.     

Strategies for mining fungal natural products

Fungi are well known for their ability to produce a multitude of natural products. On the one hand their potential to provide beneficial antibiotics and immunosuppressants has been maximized by the pharmaceutical industry to service the market with cost-efficient drugs. On the other hand identification of trace amounts of known mycotoxins in food and feed samples is of major importance to ensure consumer health and safety. Although several fungal natural products, their biosynthesis and regulation are known today, recent genome sequences of hundreds of fungal species illustrate that the secondary metabolite potential of fungi has been substantially underestimated. Since expression of genes and subsequent production of the encoded metabolites are frequently cryptic or silent under standard laboratory conditions, strategies for activating these hidden new compounds are essential. This review will cover the latest advances in fungal genome mining undertaken to unlock novel products.     

Atypical morphology of dark septate fungal root endophytes of<Emphasis Type="Italic"> Bouteloua</Emphasis> in arid southwestern USA rangelands

Native grasses of semi-arid rangelands of the southwestern USA are more extensively colonized by dark septate endophytes (DSE) than by traditional mycorrhizal fungi. Roots of dominant grasses ( Bouteloua sp.) native to arid southwestern USA rangelands were prepared and stained using stains specific for fungi (trypan blue) and for lipids (sudan IV). This revealed extensive internal colonization of physiologically active roots by atypical fungal structures that appear to function as protoplasts, without a distinguishable wall or with very thin hyaline walls that escape detection by methods staining specifically for fungal chitin. These structures were presumed to be active fungal stages that progressed to form stained or melanized septate hyphae and microsclerotia characteristic of DSE fungi within dormant roots. The most conspicuous characteristic of these fungi were the unique associations that formed within sieve elements and the accumulation of massive quantities of lipids. This interface suggests a biologically significant location for carbon transfer between the plant and fungus. The continuous intimate association with all sieve elements, cortical and epidermal cells as well as external extension on the root surface and into the soil indicates that they are systemic and considerably more prevalent than previously thought. A fungal network associated with a mucilaginous complex observed on the root surface and its potential role in root function in dry soil is discussed. It is suggested that those fungi that non-pathogenically and totally colonize plant cells be classed as systemic endophytic fungi (SEF). This would refine the broad designation of DSE fungi. The potential mutualistic benefit of SEF for native plants in arid ecosystems based on the extent of lipid accumulation and its apparent distribution is discussed.     

Survey of bioaerosol in different indoor working environments in central India

Aerobiological studies at three different indoor sitesviz., food grain godowns, library building and bakerywere carried out. The main objective of the study wasto find out the fungal flora at these places and itsimpact on the organic materials which are stored /processed there. The study reveals that over 40fungal types prevail in such organic matter richenvirons. The incidence of fungi was found to befluctuating according to months and seasons. Studieswere carried out by Andersen two stage sampler. Maximum spore concentration was observed in thelibrary followed by bakery and food grain godowns. Common spore types belonged to the species of Aspergillus. Alternaria, Cladosporium,Helminthosporium, Curvularia, Rhizopus etc. Suchfungi are mainly responsible for the deterioration ofpaper materials in library and food grains in godowns. Bakery products also get contaminated with a goodnumber of fungi types. The people working at suchplaces also get allergic due to these fungalcomponents.     

Thermophilic fungi in air samples in surroundings of compost piles of municipal,agricultural and horticultural origin

In this study, the effect of the thermophilic fungi of composts was analysed on the fungal composition of the air above. Air samples were collected with an Andersen air sampler at 1.5 m height in three large industrial composting facilities treating different waste types. Repetition was collected on three calm and rain-free days of three consecutive weeks in October 2011, in January, April and July 2012; five plates were exposed successively per sampling day. Compost samples were also collected (averaging 1 kg/compost piles). Air and compost samples were cultured at 50 °C. The thermophilic fungal composition of the air near the compost piles of different waste types differed significantly (p < 0.05) from that of the control site above a grassland ecosystem at each sampling time. Seasonal differences could be detected regarding the total number of thermophilic fungi in the air near the agricultural and horticultural compost types, but smaller differences were found near the municipal compost type. A total of 13 and 11 fungal species were detected in the compost and air samples where the dominant species were Thermomyces lanuginosus and Rasamsonia emersonii, respectively. The concentration of airborne thermophilic fungi was higher near the horticultural compost type and lower near the municipal compost. The results suggest that the differences between the incidences of some species in composts and associated aerosols refer to spore ontogeny and biological mechanisms of spore liberation.