Toxigenic fungi isolated from roquefort cheese

To evaluate the potential for mycotoxin production by fungi contaminating blue-veined cheese, as well as by the ripening fungus,Penicillium roqueforti, the fungal flora of six of local and imported brands was determined. A total of 19 fungi were isolated from the six brands tested. Fourteen of the isolates were toxic to chicken embryos. The toxigenic fungi produced the following mycotoxins:Aspergillus fumigatus, kojic acid;A. versicolor, sterigmatocystin;Penicillium roqueforti, penicillic acid and unidentified toxic metabolites.     

Penicillium glabrum cork-colonising isolates - preliminary analysis of their genomic similarity

The cork stopper manufacturing process includes an operation, known as stabilisation, by which humid cork slabs are extensively colonised by fungi. The effects of fungal growth on cork are not completely understood although they are considered to be involved in the so-called "cork taint" of wine. It is essential to (a) identify environmental constraints which define the appearance of the colonising fungal species and (b) trace their origin to the forest and/or the manufacturing space. The present article correlates two sets of data, from consecutive years and the same season, of systematic sampling of two manufacturing units, located in the North and South of Portugal. Chrysonilia sitophila dominance was confirmed, followed by a high diversity of Penicillium species. Penicillium glabrum, which was found in all samples, was the most frequently isolated species. P. glabrum intra-species variability was investigated using DNA fingerprinting techniques revealing highly discriminative polymorphic markers in the genome. Cluster analysis of P. glabrum data was discussed in relation to the geographical location of strains, and results suggest that P. glabrum arise from predominantly the manufacturing space, although cork specific fungi can contribute.     

Airborne Fungal Colony-Forming Units in Outdoor and Indoor Environments in Yokohama,Japan

The fungal concentration and flora in indoor and outdoor air in Yokohama, Japan were analyzed with a Reuter centrifugal air sampler and dichloran 18% glycerol agar (DG18), and compared with the levels assessed with potato dextrose agar (PDA). The number of fungal colony-forming units (CFU) in outdoor air was < 13–2750/m³; Cladosporium spp. predominated, followed by Alternaria spp. and Penicillium spp. The fungal concentration in outdoor air peaked in September. The concentrations of fungi in outdoor air (n = 288) were significantly correlated with the maximum temperature of the day, minimum temperature of the day, average temperature of the day, average velocity of wind of the day, average temperature of the month, average relative humidity of the month and precipitation of the month. In indoor air, the fungal CFU was < 13–3750/m³. Cladosporium spp. predominated, followed by the xerophilic fungi such as the Aspergillus restrictus group, Wallemia sebi, the A. glaucus group, and Penicillium spp. The fungal concentration in indoor air peaked in October. The concentrations of fungi in indoor air (n = 288) were significantly correlated with the indoor temperature, indoor relative humidity and the outdoor climatic factors mentioned above, except for the average velocity of wind of the day. This revised version was published online in June 2006 with corrections to the Cover Date.     

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.     

The biosynthesis of low-molecular-weight nitrogen-containing secondary metabolite-alkaloids by the resident strains of Penicillium chrysogenum and Penicillium expansum isolated on the board of the Mir space station

The analysis of the absorption spectra of the low-molecular-weight nitrogen-containing secondary metabolites--alkaloids--of 4 Penicillium chrysogenum strains and 6 Penicillium expansum strains isolated on board the Mir space station showed that all these strains synthesize metabolites of alkaloid origin (roquefortine, 3,12-dihydroroquefortine, meleagrin, viridicatin, viridicatol, isorugulosuvin, rugulosuvin B, N-acetyl-tryptamine, and a "yellow metabolite" containing the benzoquinone chromophore).     

Unveiling the fungal mycobiota present throughout the cork stopper manufacturing process

A particular fungal population is present in the main stages of the manufacturing process of cork discs. Its diversity was studied using both dependent (isolation) and independent culture methods (denaturing gel gradient electrophoresis and cloning of the ITS1-5.8S-ITS2 region). The mycobiota in the samples taken in the stages before and after the first boiling seems to be distinct from the population in the subsequent manufacturing stages. Most isolated fungi belong to the genera Penicillium, Eurotium and Cladosporium. The presence of uncultivable fungi, Ascomycota and endophytes in raw cork was confirmed by sequencing. The samples taken after the first boiling contained uncultivable fungi, but in a few samples some isolated fungi were also detected. The main taxa present in the following stages were Chrysonilia sitophila, Penicillium glabrum and Penicillium spp. All applied techniques had complementary outcomes. The main factors driving the shift in cork fungal colonization seem to be the high levels of humidity and temperature to which the slabs are subjected during the boiling process.     

Nasal fungal microbiota in allergic and healthy subjects

Environmental fungi, moulds and yeasts could reach the nasal cavity with the inhaled air causing respiratory symptoms in atopic subjects, but little is known about the fungal flora of this site. In the present study samples of the nasal cavities of 135 subjects aged 18-35 years (48 allergic patients to fungi, mites and/or cat fur and from 87 normal subjects--healthy, control group) were cultured. All of them lived in the metropolitan area of Barcelona. Fungi were isolated from 41.5% of healthy people and in 14.8% of allergy patients (p = 0.011). Morphologically, 50.4% of the isolates were located within 4 genera: Cladosporium, Penicillium, Aspergillus and Alternaria, fungi which are considered the most allergenic. The most prevalent species were: Cladosporium herbarum and C. cladosporioides (23.6%). Alternaria alternata was isolated only in 8.8% of samples from the allergic group, although most subjects were sensitive to this species. There were not differences in the isolation rate between genera and smoking-no-smoking groups. The lower prevalence of nasal fungi from allergic patients could be related to the nasal insufficiency, the hypersecretion and the larger use of handkerchiefs.     

The ability of filamentous fungi to produce acids on indoor building materials

Sixty two filamentous fungi isolated from paint coatings, wallpaper, carton-gypsum board, and indoor air in buildings were screened for acid activity. It was found that 64.5% of strains produce acids on medium with bromo-cresol purple, where 18% of the strains were distinguished by very high acid activity (acid activity coefficient Q = 1.32–2.83), including the species:Aspergillus niger, Aspergillus versicolor, Penicillium expansum, Penicillium brevicom pactum, Penicillium chrysogenum, Cladosporium cladosporioides, Stachybotrys chartarum, Mucor globosus, Ulocladium chartarum andAlternaria alternata. Research indicated that filamentous fungi considerably decrease the pH of the medium when that medium containing building material. The greatest acid production and pH decrease of the medium was observed during the growth of filamentous fungi in a medium with mortar, while the production of acids was less in a medium with cartongypsum board, gypsum, and wallpaper. Filamentous fungi produced succinic, oxalic, malic and fumaric acids in the medium with indoor building materials. It was stated that the type of building material affects the spectrum and quantity of organic acids produced by filamentous fungi.     

Bird feather fungi from Svalbard Arctic

Despite feather fungi being an important component of the Arctic fungal flora, their ecological role and diversity are not fully known. In the current study, fungal cultures were isolated from feathers (barnacle goose, common eider, and glaucous gull) collected in the Ny-Ålesund region, Svalbard. Isolates were identified by ITS region sequences, which include the ITS1, ITS2, and 5.8S rRNA. The result showed culturable yeast and filamentous fungi belonging to three classes: Ascomycota (Pyrenochaetopsis pratorum, Cladosporium herbarum, Thelebolus microsporus, Aspergillus versicolor, Penicillium commune, and Venturia sp.), Basidiomycota (Mrakia blollopis and Rhodotorula mucilaginosa), and Zygomycota (Mucor flavus). Most of the fungal isolates appeared to be cold-tolerant, and about 60 % of the isolates showed keratinase activity. The reasonably low fungal diversity colonizing feathers indicates that the birds of Svalbard are casual carriers of fungi which may result in a negligible impact on their health. To the best of our knowledge, this is the first record of fungal communities present on the feathers of birds in the high Arctic.     

Contamination of air systems: 1987–88 assessment and prospects of the Grenoble intervention group

Summary In order to evaluate bacterial and fungal contamination, the authors used a qualitative and quantitative procedure and investigated 47 air conditioning and humidifying units. Air samples were studied using Biotest RCS biocollector (160×6 liters on 6 different selective media), whilst dust samples were analysed after spreading 10 mg onto fungal media. Selective research ofLegionella and fungi were performed using water filtrates and specific media. Even though, the most frequently identified species were quite common environmental fungi, mainlyPenicillium, Alternaria andAspergillus fumigatus (36 out of 47 collections), the health effects of intense exposure to these common moulds are not really known.Staphylococcus aureus was identified 6 times and thermophilic actinomyces species (A. candidus) once. From the quantitative analysis a significant relationship was found between some risk factors and airborne contamination. Indeed bacteria and fungi number depends on the humidifying system (steam or washer), efficiency of secondary filters, relative humidity percentage (< or >60%), installation maintenance, industrial activity and complaints of people at risk. The presence of air recycling and biocide use seems only to have an influence on fungal flora. Having determined a global risk score for each air conditioning unit, the authors underline the fact that bacterial and fungal airborne contamination increases with score. Moreover, for a global, metrological, medical and technical evaluation, a multidisciplinary approach has proved itself to be indispensable.     

健康与患病刺梨植株可培养叶际真菌菌群差异比较

本研究比较了健康与患叶斑病刺梨植株叶际真菌群落特征差异,以期探索病原菌的潜在来源,为人工构建具拮抗功能群落和刺梨叶斑病的生物防控提供参考。通过可培养方法对不同健康状况的刺梨叶际真菌进行分离培养,结合形态学和分子系统学对菌株进行综合鉴定;利用FUNGuild平台对真菌进行功能注释;结合根际、根部真菌作拆分网络分析探索病原菌的潜在来源。本研究结果表明：1）刺梨叶际真菌具有丰富的多样性。从刺梨叶际8个样品中共分离到真菌266株,其隶属于3门、6纲、13目、30科、46属中的61个种。其中,健康植株叶际内生真菌（LHE）包括8属10种27株,附生真菌（LHS）包括33属37种77株。患病叶际内生真菌（LDE）分离到7属10种38株;附生真菌（LDS）分离到31属35种124株。2）不同样品的真菌优势属和特有类群有差异。不同健康状况下叶际附生真菌的优势属均为拟盘多毛孢属Pestalotiopsis,但二者的相对多度存在差异,LHS为11.49%,LDS为32.26%;内生真菌优势属二者均为链格孢属Alternaria,但相对多度各异,LHE为33.33%,LDE为63.16%。其中,LHE特有类群为盘长孢状刺盘孢 Colletotrichum gloeosporioides和果生刺盘孢Colletotrichum fructicola等8种;LDE特有类群为茄链格孢 Alternaria solani和Didymella sinensis 等8种;LHS特有类群是草酸青霉Penicillium oxalicum和Peniophora crassitunicata等21种;LDS特有类群是尖孢镰刀菌Fusarium oxysporum、赭绿青霉Penicillium ochrochloron和易脆毛霉Mucor fragilis等19种。3）不同样品叶际真菌功能不同。经FUNGuild解析表明,LHS、LHE和LDE的叶际真菌功能群主要以腐生型为主,LDS则主要以植物病原菌群为主。本研究结果初步揭示刺梨植株健康与患叶斑病叶际间真菌多样性、群落组成及营养功能群存在差异,植株健康状况与其真菌群落特征密切相关;叶斑病病原菌主要源于刺梨叶际的附生微生物群。     

肠道真菌研究进展

摘要：肠道真菌是肠道微生物的重要组成部分，在肠道正常菌群中所占比例极小，主要包括假丝酵母属（Candida）、隐球酵母属（Cryptococcus）、青霉属（Penicillium）、曲霉属（Aspergillus）、红酵母属（Rhodotorula）等，对维持肠道微生态稳态和机体健康具有重要作用，也与抗生素相关性腹泻（AAD）、乙型肝炎、炎症性肠病（IBD）等疾病的发病机制息息相关。本文主要总结了目前已发现的肠道真菌属及其对人体的影响机制以及肠道真菌相关的研究方法，为肠道真菌在疾病的诊断和治疗方面的进一步研究提供依据与思路。     

Abundance of airborne Penicillium CFU in relation to urbanization in Mexico City.

Air was sampled simultaneously at three localities in Mexico City differing in urbanization index and air pollution level on 22 days during a period covering both dry and rainy seasons. An Andersen two-stage microbial sampler was used for 15 min at 28 liters min-1 to isolate culturable fungi on malt extract agar. After exposure, plates were incubated at 25 degrees C for 48 to 72 h before colonies were counted and identified to give concentrations of total fungal spores and of Penicillium spp., expressed as CFU per cubic meter of air. Total fungi numbered 91 to 602 CFU m-3 in Tlalpan Borough (southern area), 40 to 264 CFU m-3 in Cuauhtémoc Borough (downtown), and 26 to 495 CFU m-3 in Gustavo A. Madero Borough (northern area). Although Penicillium spp. were the second most frequently isolated fungal genus, concentrations were small, with a maximum of only 133 CFU m-3. Twice as many colonies were isolated in the southern area, with an urbanization index of 0.25 (arithmetic mean, 41 CFU m-3), as at other sampling stations with greater urbanization indices (arithmetic means, 19 and 20 CFU m-3). In the downtown area, with an urbanization index of 1.0, Penicillium spp. were more numerous than any other genus and formed 25% of the total fungal count compared with 14 and 17% in the other areas. Concentrations of airborne Penicillium spp. did not differ significantly between rainy and dry seasons.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effectiveness of imazalil to control the effect of fungal deterioration on mummies at the Mexico City Museum "El Carmen"

We present a study on the control and elimination of the fungi affecting the mummies specifically at the museum "El Carmen", in San Angel, Mexico City. Twelve analysed mummies presented an important deterioration attributed to colonizing fungi. The degree of fungal contamination and the efficacy of imazalil were evaluated. Two samplings were performed in order to isolate and identify the fungal genera, one for control and the other after the treatment. Isolation was done by the carpet-square technique and identification was performed by morphological features. Each sampling gave a total of 100 samples as follows: 17 from the air, 23 from the walls and 60 from the mummies. Samples were cultured on Sabouraud dextrose agar. From the first sampling a total of 649 colonies corresponding to 24 genera were obtained being the most frequent Penicillium, Cladophialophora and Aspergillus. From the second sampling, after the imazalil treatment, which was applied by means of lit candles containing the antifungal drug, 57 colonies were recovered, representing a 91.2% fungal reduction; 18 genera were eliminated. In spite of resistance showed by many Penicillium strains, the imazalil is an alternative drug for the control of fungal colonization on these studied materials.     

Viable fungi in the air of indoor environments of the National Archive of the Republic of Cuba

The aim of this study was to analyze the viable fungi in the air of indoor environments of the National Archive of the Republic of Cuba (NARC). The samples were taken in fifteen areas (five points for the three floors) at 2 days (rainy and dry seasons) using a biocolector SAS in 3 h: 10:00 a.m., 1:00 p.m. and 4:00 p.m. The fungal concentration was indicated as colony forming units per cubic meter of air (cfu/m³ of air). It was concluded that indoor air of the NARC was not polluted due to the average fungal concentration that was obtained (59 and 327 cfu/m³ of air in rainy and dry seasons, respectively). In front of the electric transformers, the fungal concentration was significantly higher than those obtained along the corridors of the lower-ground floor. A trend to the increasing of fungal concentration was observed: when the floors were increased, in dry season was higher than rainy season, where also there was biggest fungal diversity. Among the fungal genera isolated, Aspergillus had the highest relative frequency of appearance, and in dry season, it was isolated Actinobacillus sp. (Actinomycetes). Six fungal species were isolated in both samplings: Aspergillus alliaceus, Aspergillus niger, Cladosporium cladosporioides, Fusarium oxysporum, Penicillium chrysogenum and Mucor racemosus. All of fungal species isolated have high biodeteriogenic capacity, and the 61 % showed pathogenic attributes.     

The system approach to the identification of aflatoxigenic fungi in foodstuffs and feedstuffs

Aspergillus flavus, A parasiticus, A nomius, A tamarii andA pseudotamarii are important microorganisms capable of producing aflatoxins and further mycotoxins. Aflatoxigenic Aspergillus species are morphologically similar species belonging to the Aspergillus section Flavi. The aflatoxigenic fungal strains were isolated from foods (cereals, pulses, oilseeds, dried fruit, spices), soil, air and water. Mycological analyses are based on valid standards and recommendations of the International Commission for Food Mycology (ICFM). The identification of isolated aflatoxigenic fungi in foodstuffs and feedstuffs can be proved by using classical mycological cultivation methods, diagnostic nutrient media, chemotaxonomy and molecular biological methods (PCR). The system approach to the identification of aflatoxigenic fungi combines these four methods. Thirty strains of the aflatoxigenic fungi were tested.     

Profiles of airborne fungi in buildings and outdoor environments in the United States

We examined 12,026 fungal air samples (9,619 indoor samples and 2,407 outdoor samples) from 1,717 buildings located across the United States; these samples were collected during indoor air quality investigations performed from 1996 to 1998. For all buildings, both indoor and outdoor air samples were collected with an Andersen N6 sampler. The culturable airborne fungal concentrations in indoor air were lower than those in outdoor air. The fungal levels were highest in the fall and summer and lowest in the winter and spring. Geographically, the highest fungal levels were found in the Southwest, Far West, and Southeast. The most common culturable airborne fungi, both indoors and outdoors and in all seasons and regions, were Cladosporium, Penicillium, nonsporulating fungi, and Aspergillus. Stachybotrys chartarum was identified in the indoor air in 6% of the buildings studied and in the outdoor air of 1% of the buildings studied. This study provides industrial hygienists, allergists, and other public health practitioners with comparative information on common culturable airborne fungi in the United States. This is the largest study of airborne indoor and outdoor fungal species and concentrations conducted with a standardized protocol to date.     

Mites and fungi in heavily infested stores in the Czech Republic

Toxigenic and allergen-producing fungi represent a serious hazard to human food and animal feed safety. Ninety-four fungal species were isolated from mite-infested samples of seeds taken from Czech seed stores. Fungi were isolated from the surface of four kinds of seeds (wheat, poppy, lettuce, and mustard) and from the gut and external surface of five species of mites (i.e., Acarus siro L., 1758, Caloglyphus rhizoglyphoides (Zachvatkin, 1973), Lepidoglyphus destructor (Schrank, 1781), Tyrophagus putrescentnae (Schrank, 1781) and Cheyletus malaccensis Oudemans 1903) separately. Multivariate analysis of fungi complex composition showed that the frequency of fungal was species significantly influenced by the kind of seed. Fungal frequencies differed between mites gut and exoskeleton surface and between the surfaces of mites and seeds. Three groups of fungal species were recognized: 1) mite surface-associated fungi: Penicillium brevicompactum, Alternaria alternata, and Aspergillus versicolor; 2) mite surface- and seed-associated fungi: Aspergillus niger, Penicillium crustosum, Penicillium aurantiogriseum, Penicillium chrysogenum, and Aspergillus flavus; and 3) seed-associated fungi: Cladosporium herbarum, Mucor dimorphosporus f. dimorphosporus, Botrytis cinerea, Penicillium griseofulvum, and Eurotium repens. Mite-carried species of microfungi are known to produce serious mycotoxins (e.g., aflatoxin B1, cyclopiazonic acid, sterigmatocystin, ochratoxin A, and nephrotoxic glycopeptides) as well as allergen producers (e.g., A. alternata and P. brevicompactum). Storage mites may play an important role in the spread of some medically hazardous micromycetes. In addition, these mite-fungi associations may heighten the risk of occurrence of mycotoxins in food and feed stuffs and cause mixed contamination by fungal and mite allergens.     

Osmophilous fungi in the salt marshes of Kuwait.

During an investigation of the mycoflora inhabiting saline soils in Kuwait, special attention was focused on osmophilous fungi. A total of 101 species belonging to 46 genera were encountered from 40 soil samples collected from salt marshes using Czapek's agar supplemented with 40% sucrose. Soil samples were collected from different habitats at different distances from the water edge. Soils near to the water edge were poor in their fungal content, while those taken from areas covered by Juncus arabicus contained highest fungal populations. The recorded genera were classified as follows: 7 were of high frequency of occurrence, 8 moderate, 17 low, and 13 were rare. The order of dominance was Aspergillus, Alternaria, Penicillium, Cephalosporium, Fusarium, Stachybotrys, and Drechslera. Comparison between our results and those in other studies showed that there is no fungal flora characteristic of saline soils.