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.     

Secretion of β-galactosidases by Aureobasidium pullulans and Penicillium brevicompactum on polygalacturonate medium

An assay has been developed to detect production of extracellular β -galactosidases by fungi during growth on a defined medium containing polygalacturonate. Using this method strains of the yeast-like fungus Aureobasidium pullulans and the filamentous fungus Penicillium brevicompactum which secrete β -galactosidase have been isolated. Seventy strains of yeast were tested but none secrete detectable extracellular β -galactosidase during growth on polygalacturonate agar medium.     

Mycoflora and mycotoxins of Brazilian cashew kernels

Kernel samples of common and dwarf Brazilian cashew nuts were highly contaminated with field and storage fungi in comparison to healthy ones. In general, dwarf cashews were more contaminated than common. A total of 37 fungal species were identified. Aspergillus niger was the dominant species with more colonies being isolated from dwarf kernels. A. flavus was the next most frequently isolated species. Penicillium brevicompactum, and P. glabrum were the most frequently isolated penicillia, with higher contamination recorded from dwarf kernels. Chaetomium globosum was recorded at a high level. Nine species were recorded from cashew kernels for the first time. Multimycotoxin analysis by tlc and hplc were positive for mycotoxins and other secondary metabolites particularly from the infected samples. Hplc was only carried out on dwarf cashews. Aflatoxins were not detected by quantitative high performance thin layer chromatography. This revised version was published online in June 2006 with corrections to the Cover Date.     

A potential resource for bioconversion of domestic wastewater sludge

Twenty seven filamentous fungal strains representing five genera; Aspergillus, Penicillium, Trichoderma, Myriodontium and Pleurotus were isolated from four sources; domestic wastewater sludge cake (SC) from IWK (Indah Water Konsortium) wastewater treatment plant, palm oil mill effluent compost from Sri Ulu palm Oil Processing Mill, compost of plant debris, and fungal fruiting bodies from a rotten wood stump. Thirty-three strains/isolates were tested for their ability to convert domestic wastewater sludge into compost by assessing biomass production and growth rate on sludge enriched media. The strains/isolates Aspergillus niger, SS-T2008, WW-P1003 and RW-P1 512 produced the highest dry biomass at higher sludge supplemented culture media from their respective group (Aspergillus, Trichoderma, Penicillium and Basidiomycetes, respectively). This implied these strains are better adapted for growth at higher sludge rich substances, and subsequently may be efficient in bioconversion/biodegradation of sludge. The fungi isolated from ecological closely related sources were more amendable to adaptation in a sludge rich culture media.     

Screening and identification of a Penicillium brevicompactum strain isolated from the fruiting body of Inonotus obliquus and the fermentation production of mycophenolic acid

Mycophenolic acid (MPA) is a fungal metabolite with a variety of biological activities and widely applied in clinical practices. We herein aimed to isolate a new Penicillium brevicompactum strain from the fruiting body of Inonotus obliquus to improve the production of MPA. The fruiting body of Inonotus obliquus was used to isolate P. brevicompactum strains. Identification of the P. brevicompactum strain was performed by sequencing and phylogenetic tree analysis. HPLC assay was conducted to identify the production of MPA. Submerged liquid fermentation and bi-directional fermentation were applied to improve the yield of MPA. The candidates of the P. brevicompactum strain were isolated and screened from the fruiting body of Inonotus obliquus collected from Changbai mountains in China. Based on sequencing and phylogenetic tree analysis of 18S rDNA-ITS, the strain MC-4 was finally identified as P. brevicompactum. And HPLC assay indicated that the isolated P. brevicompactum strain could produce MPA in metabolic products. The optimized conditions of submerged liquid fermentation were as follows: 100 g/L Chinese yam in a liquid PDB medium, pH 6, fermentation temperature of 24 °C, shaker speed of 130 r/min, and fermentation time of 6 days. The maximum value of MPA production was 1.415 g/L after submerged liquid fermentation. Furthermore, the yield of MPA could be significantly increased to 1.537 g/L after bi-directional fermentation with the extractive from fructus Swietenia macrophylla (FSM). We demonstrated that a Penicillium brevicompactum strain isolated from the fruiting body of Inonotus obliquus can be used to improve the production of MPA by submerged liquid fermentation and bi-directional fermentation. This would provide a novel approach for more efficient and safer production of MPA.     

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.     

Antagonism between Bacteria and Fungi on Decomposing Aquatic Plant Litter

Bacterial and fungal decomposers of aquatic plant litter may exhibit either synergistic or antagonistic interactions, which are likely to influence microbial growth as well as the decomposition of litter and, eventually, the carbon metabolism of aquatic systems. To elucidate such interactions, we inoculated decomposing Phragmites culms in microcosms with fungal isolates and with natural communities of bacteria and fungi in different combinations. The development of fungal and bacterial biomass and the carbon dynamics were studied during several months of degradation. The results show a bilateral antagonistic relationship between bacteria and fungi. After 3 months, fungal biomass accumulation was approximately 12 times higher in the absence than in the presence of bacteria. Bacterial biomass accumulation was about double in the absence of fungi compared to when fungi were present. Similar interactions developed between a natural assemblage of bacteria and five different fungal strains isolated from Phragmites litter (three identified hyphomycetes and two unidentified strains). Despite the great difference in biomass development between the treatments, the carbon metabolism was similar regardless of whether fungi and/or bacteria were present alone or in coexistence. We suggest that the antagonism between bacteria and fungi is an important controlling factor for microbial colonization and growth on aquatic plant litter.     

A Gene Probe for the Patulin Metabolic Pathway with Potential for Use in Patulin and Novel Disease Control

The iso-epoxy dehydrogenase gene of the patulin metabolic pathway was detected in environmental samples, Penicillium expansum and P. brevicompactum isolated from an organic orchard. Patulin was not detected from P. brevicompactum . Both traits were negative for other penicillia. In general, control of disease and mycotoxin reduction will be optimized only if all sites of infection and contamination are targeted.     

Penicillium astrolabium and Penicillium neocrassum, two new species isolated from grapes and their phylogenetic placement in the P. olsonii and P. brevicompactum clade

We describe two new terverticillate Penicillium species isolated from grapes on the basis of phenotypic and phylogenetic differences from known species. The strains were isolated in the course of a study to establish the mycobiota of grapes in Portugal. Penicillium astrolabium is phenotypically similar to P. olsonii but differs from it by two cultural characters, growth rates and the colony reverse color. P. neocrassum is similar to P. brevicompactum but is readily distinguished by sclerotia production. Phylogenetically P. astrolabium and P. neocrassum are placed respectively in the P. olsonii and P. brevicompactum clade. Multilocus analysis confirmed the genetic distinctiveness of both species. The parsimony trees obtained for ITS-lsu rDNA region and two protein coding genes, calmodulin and beta-tubulin, show congruence for all the species in the Olsonii series: P. brevicompactum, P. bialowiezense, P. olsonii, P. astrolabium and P. neocrassum, indicating that these taxa are genetically well isolated.     

Dynamics in the microbiology of maize silage during whole‐season storage

Aims: To monitor seasonal variations in the microbiology of maize silage and to determine whether the risk of fungal spoilage varies during whole‐year storage. Methods and Results: A continuous survey of 20 maize silage stacks was conducted over a period from three to 11 months after ensiling. Filamentous fungi, yeasts and lactic acid bacteria (LAB) were enumerated at five time‐points, and cultivable species of filamentous fungi were identified. Significant differences in the numbers of filamentous fungi, yeast and LAB were detected. The highest numbers of fungi were five to seven and the lowest 11 months after ensiling, while the LAB decreased in numbers during the study. Filamentous fungi were isolated from all stacks at all time‐points. The most abundant toxigenic mould species were Penicillium roqueforti, Penicillium paneum and Aspergillus fumigatus. Conclusions: There are significant variations in the microbiology of maize silage over a whole storage season. The risk of fungal spoilage was highest 5–7 months after ensiling and lowest after 11 months. Significance and Impact of the Study: This information is valuable in the assessment of health risks connected with spoiled maize silage and may be useful in the management of maize silage stacks, when whole‐season storage is applied.     

Fungi isolated from asthmatic homes in the Taipei area.

Indoor fungi in Taipei were surveyed during 1969-1974 using Sabouraud's agar plates. No remarkable yearly variation of fungal incidence was observed. The fungal population was found to be different from home to home. The dominant members were Aspergillus, Penicillium, Mycelia sterila, Oospora, Rhodotorula, Hormodendrum and yeasts. The order of frequency of fungi found indoors was quite different from that of outdoor fungi. The fungal spores were more numerous during summer months, and this trend was more apparent for Aspergillus, Oospora, Rhodotorula, and Hormodendrum. The kind and the incidence of fungi found in asthmatic homes were not different from those detected in non-asthmatic homes, except that in asthmatic homes there were greater numbers of fungi.     

Predictive modelling approach applied to spoilage fungi: growth of Penicillium brevicompactum on solid media

Growth of Penicillium brevicompactum was examined on five solid media. Fungal growth was established by diameter measurements up to 50 days. Seventy experimental curves were fitted by Baranyi's primary predictive model. The growth rates were then analysed by non-parametric statistical methods. Penicillium brevicompactum could colonize the surface of solid media containing up to 700 g l-1 of sugar (50% glucose-50% fructose) with a growth rate of 0.9 mm day-1 (median values). Fitting curves by non-linear models followed by a non-parametric multiple comparison seems to be a convenient method for detecting differences in fungal growth on solid media. These two methods would be useful for studying fungal spoilage of bakery products with intermediate water activity.     

Fungi associated with black tea and tea quality in the Sultanate of Oman

Forty-eight samples of four popular commercial brands of black tea ( Camellia sinensis L.) were purchased from the local markets in Muscat area, Sultanate of Oman. Tea leaves were surveyed for mycoflora. Five fungal species were isolated with A. niger as the most dominant in all the brands having percentage contamination ranging between 0.66% and 30.34%. Other fungi isolated were Aspergillus flavus, Penicillium spp. and Pacelomyces spp. but having average percentages of 0.6%, 0.84% and 0.21% respectively. Significant differences were found among the batches contaminated by A. niger. None of the 25 A. flavus strains screened for aflatoxins were found aflatoxigenic. The total ash, water-soluble ash, and mineral concentration of the samples were within the British standards and were not affected by fungal contamination. The results showed that black tea is contaminated by fungi that might constitute health hazards for humans. The post harvest contamination of tea could be eliminated or reduced if processing is conducted under more hygienic conditions. This revised version was published online in June 2006 with corrections to the Cover Date.     

Isolation and evaluation of tannin-degrading fungal strains from the Mexican desert

Eleven fungal strains (4 Penicillium commune, 2 Aspergillus niger, 2 Aspergillus rugulosa, Aspergillus terricola, Aspergillus ornatus and Aspergillus fumigatus) were isolated, characterized morphologically and by their capacity to degrade tannins. Aspergillus niger Aa-20 was used as control strain. Several concentrations of hydrolysable tannin (tannic acid) were used as sole carbon source. All strains were able to degrade hydrolysable tannins. Aspergillus niger GH1 and PSH showed the highest tannin-degrading capacity (67 and 70%, respectively). Also, the fungal capacity to degrade condensed tannin (catechin) was tested. Aspergillus niger PSH and Penicillium commune EH2 degraded 79.33% and 76.35% of catechin. The results demonstrated the capacity of fungi to use hydrolysable and condensed tannins as carbon source.     

Activity of acid and alkaline phosphatases (intracellular and extracellular) in rhyzosphere fungi from Arachis hypogaea (Papiloneaceae)

The potential role of the fungi, isolated from the peanut rhizosphere, in the production of extracellular and intracellular acid and alkaline phosphatase, was evaluated in vitro. Acid and alkaline extracellular phosphatases showed the highest activities, and the Penicillium species were the most efficient in their production. The correlation analysis showed that extracellular acid and intracellular acid phosphatase produced by Aspergillus niger A. terreus, Penicillium sp. y P. brevicompactum were negatively correlated; while the extracellular and intracellular phosphatase activities, were positively correlated. The extracellular acid phosphatase activities produced in vitro by majority of fungi assayed, were not correlated with the acid phosphatase activity present in the peanut soil rhizosphere. Nevertheless, the extracellular alkaline phosphatase activities produced in vitro, were negatively correlated with the extracellular alkaline phosphatase activities present in the rhizosphere. The ability of phosphatase production by fungi isolated from peanut rhizosphere suggests they have great potential to contribute to the P mineralization in this zone.     

Plant and Fungal Diversity in Gut Microbiota as Revealed by Molecular and Culture Investigations

Background

Few studies describing eukaryotic communities in the human gut microbiota have been published. The objective of this study was to investigate comprehensively the repertoire of plant and fungal species in the gut microbiota of an obese patient.

Methodology/Principal Findings

A stool specimen was collected from a 27-year-old Caucasian woman with a body mass index of 48.9 who was living in Marseille, France. Plant and fungal species were identified using a PCR-based method incorporating 25 primer pairs specific for each eukaryotic phylum and universal eukaryotic primers targeting 18S rRNA, internal transcribed spacer (ITS) and a chloroplast gene. The PCR products amplified using these primers were cloned and sequenced. Three different culture media were used to isolate fungi, and these cultured fungi were further identified by ITS sequencing. A total of 37 eukaryotic species were identified, including a Diatoms (Blastocystis sp.) species, 18 plant species from the Streptophyta phylum and 18 fungal species from the Ascomycota, Basidiomycota and Chytridiocomycota phyla. Cultures yielded 16 fungal species, while PCR-sequencing identified 7 fungal species. Of these 7 species of fungi, 5 were also identified by culture. Twenty-one eukaryotic species were discovered for the first time in human gut microbiota, including 8 fungi (Aspergillus flavipes, Beauveria bassiana, Isaria farinosa, Penicillium brevicompactum, Penicillium dipodomyicola, Penicillium camemberti, Climacocystis sp. and Malassezia restricta). Many fungal species apparently originated from food, as did 11 plant species. However, four plant species (Atractylodes japonica, Fibraurea tinctoria, Angelica anomala, Mitella nuda) are used as medicinal plants.

Conclusions/Significance

Investigating the eukaryotic components of gut microbiota may help us to understand their role in human health.     

植物病原真菌产生ABA的初步研究

根据植物病理学和微生物生态学原理,建立了脱落酸（ＡＢＡ）产生菌的筛选模型。从不同的植物寄主上分离到３６株真菌,其中有十余株菌株能产生天然的脱落酸,这些真菌分别属于葡萄孢属（Ｂｏｔｒｙｔｉｓ）、青霉属（Ｐｅｎｉｃｉｌｉｕｍ）、根霉属（Ｒｈｉｚｏｐｕｓ）和镰孢属（Ｆｕｓａｒｉｕｍ）。其中一株ＡＢＡ的产量达到４６ｇ／ｇ培养基,经鉴定为灰葡萄孢（Ｂ．ｃｉｎｅｒｅａ）。在此基础上,对该菌株的培养条件进行了优化。     

Detection of fungal metabolites showing toxic activity through Artemia salina bioassay

The aim of this study was to detect toxic metabolites from fungi contaminating food and medicinal herbs by applying the toxicity assay to Artemia salina. According to toxicity percentages, the extracts were classified as nontoxic (NT), slightly toxic (ST), toxic (T) and highly toxic (HT). Those classified as T and HT were assayed for mycotoxins. Only 6 out of 71 strains were found to be T (8.5%) for A. salina. Penicillium brevicompactum Dierckx, isolated from sausages, was found to be HT, mainly due to the presence of ochratoxin A and two other unidentified metabolites.     

Isolation,Identification and Screening of Manganese Solubilizing Fungi From Low-Grade Manganese Ore Deposits

The present investigation reports the isolation, molecular identification and screening of manganese (Mn) solubilizing fungal strains from low-grade Mn mine tailings. Six morphologically distinct Mn solubilizing fungal strains were isolated on MnO₂-supplemented agar plates with Mn concentration of 0.1% (w/v). The biochemical characterization of the isolated fungal strains was carried out. The molecular identification by internal transcribed spacer (ITS) sequencing identified the strains as Aspergillus terreus, Aspergillus oryzae, Penicillium sp., Penicillium sp., Penicillium daleae and Penicillium sp. with GenBank accession numbers KP309809, KP309810, KP309811, KP309812, KP309813 and KP309814, respectively. The ability of the isolated fungal strains to tolerate and solubilize Mn was investigated by subculturing them on Mn-supplemented plates with concentration ranging from 0.1 to 0.5% (w/v). Mn solubilizing ability of the fungal isolates is possibly due to the mycelia production of biogenerated organic acids such as oxalic acid, citric acid, maleic acid and gluconic acid as revealed by ion chromatography. Our investigation signifies the role of fungi in biotransformation of insoluble Mn oxide.     

The relationship between a leaf-rolling moth (Dactylioglypha tonica) and fungi covering the cocoon.

To discover the relationship between a leaf-rolling moth and the fungi densely covering its cocoons, the rolled nest leaves were collected in two districts in Japan and antibacterial properties of the fungi were examined. Cocoons and fungi isolated from the nest were classified into 5 categories by the growth stages of the insects, and 7 categories based on taxonomic properties and pigment productivity, respectively. The dominant genus was Penicillium in each location. However, the composition of the fungal categories was different and seemed to depend on their circumstances. From all cocoons with larvae, the strains that belonged to the same fungal category and produced the same antibiotic (deoxyherqueinone) were isolated. From these results, the species-specific relationship between the insect and fungi or fungal products was considered to be not extremely tight, and it was suggested the period of the larval spinning of the cocoon is a key stage of this unique relationship.