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Bacterial diversity in spent mushroom compost assessed by amplified rDNA restriction analysis and sequencing of cultivated isolates.

Spent mushroom compost (SMC) is the residual by-product of commercial Agaricus spp. cultivation, and it is mainly composed of a thermally treated cereal straw/animal manure mixture colonized by the fungal biomass. Research on the valorization of this material is mainly focusing on its use as soil conditioner and plant fertilizer. An investigation of the bacterial diversity in SMC was performed using molecular techniques in order to reveal the origin of SMC microflora and its potential effect on soil microbial communities after incorporation into agricultural soils. The bacterial population was estimated by the plate count method to a mean of 2.7 10(9) colony forming units (cfu) per g of dry weight, while the numbers of Gram-positive and Gram-negative bacteria were 1.9 10(9) and 4.9 10(8) cfu per g dw respectively as estimated by enumeration on semi-selective media. Fifty bacterial isolates were classified into 14 operational taxonomic units (OTUs) following ARDRA-PCR of the 16S rDNA gene. Sequencing of the 16S rDNA amplicon assigned 12 of the 14 OTUs to Gram-positive bacteria, associated with the genera Bacillus, Paenibacillus, Exiguobacterium, Staphylococcus, Desemzia, Carnobacterium, Brevibacterium, Arthrobacter and Microbacterium of the bacterial divisions Firmicutes and Actinobacteria. Two bacterial groups have phylogenetic links with the genera Comamonas and Sphingobacterium, which belong to beta-Proteobacteria and Bacteroidetes respectively. Two potentially novel bacteria are reported, which are associated with the genera Bacillus and Microbacterium. Most of the bacteria identified are of environmental origin, while strains related to species usually isolated from insects, animal and clinical sources were also detected. It appears that bacterial diversity in SMC is greatly affected by the origin of the initial material, its thermal pasteurization treatment and the potential unintended colonization of the mushroom substrate during the cultivation process.

Agaricus↗

The white-line-in-agar test is not specific for the two cultivated mushroom associated pseudomonads, Pseudomonas tolaasii and Pseudomonas "reactans".

A sharply defined white line in vitro forms between the pathogenic form of Pseudomonas tolaasii and another Pseudomonas bacterium, referred to as "reactans". This interaction has been considered as highly specific. However, results presented in this study rise doubt about the strict specificity of this interaction, as some other pseudomonads, associated with the cultivated mushroom Agaricus bisporus, also yielded a white line precipitate when they were streaked towards Pseudomonas tolaasii LMG 2342T. Moreover, some Finnish isolates inducing brown blotch symptoms on mushrooms like P. tolaasii(T), produced a typical white precipitate when streaked towards P. "reactans" LMG5329, even though phenotypical and genotypical features exclude these isolates from the species P. tolaasii. We propose that the white-line-in-agar (WLA) test should no longer be considered as an unequivocal diagnostic trait of P. tolaasii.

Agaricales↗

Evidence for genotypic differences between the two siderovars of Pseudomonas tolaasii, cause of brown blotch disease of the cultivated mushroom Agaricus bisporus.

Sixteen representative isolates of Pseudomonas tolaasii, the causal agent of brown blotch of the cultivated mushroom Agaricus bisporus, were previously assigned to two siderovars (sv1 and sv2) on the basis of pyoverdines synthesized. Each isolate was pathogenic and produced a typical white line precipitate when cultured adjacent to Pseudomonas "reactans" strain LMG 5329. These 16 isolates of P. tolaasii, representing sv1 and sv2, were further characterized using genotypic methods to examine the relationships between the isolates. Rep-PCR studies revealed two distinct patterns from these isolates, which were consistent with the siderovar grouping. Ribotyping differentiated P. tolaasii LMG 2342T (sv1) and PS 3a (sv2) into two distinct ribotypes. A pair of primers, targeted to a 2.1-kb fragment of tl1 (encoding a tolaasin peptide synthetase), yielded the same PCR product from P. tolaasii LMG 2342T (sv1) and PS 22.2 (sv1), but not from PS 3a (sv2). Southern blot analysis indicated that homologues of tl1 are present in PS 3a, but the pattern of hybridization differed from PS 22.2 and LMG 2342T. Sequence determination and analysis of the internally transcribed spacer region ITSI for P. tolaasii LMG 2342T, LMG 6641, and PS 3a strains further supported the presence of the two siderovars. It is concluded that considerable genotypic differences exist among Finnish isolates of P. tolaasii causing brown blotch disease on the cultivated mushroom, which is in agreement with the phenotypic diversity highlighted through previous siderotyping studies.

Agaricus↗

Cyclooxygenase inhibitory and antioxidant compounds from the fruiting body of an edible mushroom, Agrocybe aegerita.

In the search for bioactive natural products from edible mushrooms, we have investigated the fruiting body of Agrocybe aegerita. The methanol extract of this mushroom yielded a fatty acid fraction (FAF), along with palmitic acid (1), ergosterol (2), 5,8-epidioxy-ergosta-6,22-dien-3beta-ol (3), mannitol (4) and trehalose (5). The composition of FAF was confirmed by GC-MS and by comparison to the retention values of authentic samples of palmitic, stearic, oleic and linoleic acids. The structures of 1-5 were established using spectroscopic methods. FAF and compounds 1-3 showed cyclooxygenase (COX) enzyme inhibitory and antioxidant activities. The inhibition values of liposome peroxidation by FAF, compounds 1 and 2 at 100 microg/ml were 75, 45, and 43%, respectively. The inhibition values of COX-I enzyme by FAF and 1-3 at 100 microg/ml were 80, 39, 19, and 57%, respectively. Similarly, COX-II enzyme activity was reduced by FAF and 1-3 at 100 microg/ml with values of 88, 45, 28, and 22%, respectively. Compounds 1, 3 and fatty acids were isolated here for the first time from the fruiting body of A. aegerita.

Agaricales↗

Tyrosinase extract from Agaricus bisporus mushroom and its in natura tissue for specific phenol removal.

Phenols are toxic pollutants found in industrial wastes imposing several risks to human health. Tyrosinase (EC 1.14.18.1) is an oxygenase oxyreductase found in several life forms, like the mushroom Agaricus bisporus. This enzyme is readily available from this fungal tissue leading to high activity extracts without extensive purification, thus suggesting its potential as a biocatalyst for applications involving biomodification of phenols or bioremediation of phenol-polluted waters. The purpose of this work was to employ a crude extract from the Agaricus bisporus mushroom and its biomass for the removal of phenol from polluted water. Experiments were carried out without pH control. The initial phenol concentration in all solutions was 100 mg l(-1). Four enzymatic concentrations (50, 100, 200 and 400 U ml(-1)) were tested. Reactions, with 200 U ml(-1) and 400 U ml(-1) enzymatic activity, led to 90% of phenol removal. Chitosan was used as a coagulant, but no significant improvement was observed. The in natura fungi was also able to remove 90% of phenol, demostrating its viability as a biocatalyst in bioremediation process.

Agaricus↗

Food for two seasons: culinary uses of non-cultivated local vegetables and mushrooms in a south Italian village.

The use of non-cultivated plants in a daily diet based on local cuisines is potentially of considerable interest to nutritional scientists, because of the plants' role as local products and their potential as sources of novel nutraceuticals. In many Mediterranean regions these traditions are at risk of disappearing, hence the urgent need to study such knowledge systems. Accordingly, an ethnobotanical survey was carried out among the 850 inhabitants of the village of Castelmezzano, in central Lucania, which is located in the inland southern Italy. Seventy-five taxa of non-cultivated and semi-cultivated local food plants and mushrooms were documented, and uncommon food uses of a few species were reported for the first time. These include Bellavalia romana, Lepista nebularis and Onopordum illyricum. Most of the recorded non-cultivated food plants and mushrooms are cooked in oil or fat. Very few are consumed raw. This article discusses in detail the traditional culinary uses of these plants, their seasonality, ethnoecology, and their economic and nutritional potentials. The article also demonstrates how food agro-biodiversity is inextricably connected with cultural heritage.

Adult↗

Radiation preservation of foods of plant origin. Part VI. Mushrooms, tomatoes, minor fruits and vegetables, dried fruits, and nuts.

In this concluding article in the series on the technological feasibility of ionizing radiation treatment for shelf life improvement of fruits and vegetables, the present status of research on several commodities that have not been dealt with earlier is discussed. The commodities include mushrooms, tomatoes, pineapples, lychees, longans, rambutans, mangostenes, guavas, sapotas, loquats, ber, soursops, passion fruits, persimmons, figs, melons, cucumbers, aubergines, globe artichokes, endives, lettuce, ginger, carrots, beet roots, turnips, olives, dates, chestnuts, almonds, pistachios, and other dried fruits and nuts. Changes induced by irradiation on metabolism, chemical constituents, and organoleptic qualities are considered while evaluating the shelf life. The commodities have been grouped into those showing potential benefits and those not showing any clear advantages from radiation treatment. Shelf life improvement of mushrooms and insect disinfestation in dried fruits, nuts, and certain fresh fruits appears to have immediate potential for commercial application.

Ascorbic Acid↗

Multiple sex pheromones and receptors of a mushroom-producing fungus elicit mating in yeast.

The mushroom-producing fungus Schizophyllum commune has thousands of mating types defined, in part, by numerous lipopeptide pheromones and their G protein-linked receptors. Compatible combinations of pheromones and receptors encoded by different mating types regulate a pathway of sexual development leading to mushroom formation and meiosis. A complex set of pheromone-receptor interactions maximizes the likelihood of outbreeding; for example, a single pheromone can activate more than one receptor and a single receptor can be activated by more than one pheromone. The current study demonstrates that the sex pheromones and receptors of Schizophyllum, when expressed in Saccharomyces cerevisiae, can substitute for endogenous pheromone and receptor and induce the yeast pheromone response pathway through the yeast G protein. Secretion of active Schizophyllum pheromone requires some, but not all, of the biosynthetic machinery used by the yeast lipopeptide pheromone a-factor. The specificity of interaction among pheromone-receptor pairs in Schizophyllum was reproduced in yeast, thus providing a powerful system for exploring molecular aspects of pheromone-receptor interactions for a class of seven-transmembrane-domain receptors common to a wide range of organisms.

ATP-Binding Cassette Transporters↗

Foraging behaviour in Drosophila larvae: mushroom body ablation.

Drosophila larvae and adults exhibit a naturally occurring genetically based behavioural polymorphism in locomotor activity while foraging. Larvae of the rover morph exhibit longer foraging trails than sitters and forage between food patches, while sitters have shorter foraging trails and forage within patches. This behaviour is influenced by levels of cGMP-dependent protein kinase (PGK) encoded by the foraging (for) gene. Rover larvae have higher expression levels and higher PGK activities than do sitters. Here we discuss the importance of the for gene for studies of the mechanistic and evolutionary significance of individual differences in behaviour. We also show how structure-function analysis can be used to investigate a role for mushroom bodies in larval behaviour both in the presence and in the absence of food. Hydroxyurea fed to newly hatched larvae prevents the development of all post-embryonically derived mushroom body (MB) neuropil. This method was used to ablate MBs in rover and sitter genetic variants of foraging to test whether these structures mediate expression of the foraging behavioural polymorphism. We found that locomotor activity levels during foraging of both the rover and sitter larval morphs were not significantly influenced by MB ablation. Alternative hypotheses that may explain how variation in foraging behaviour is generated are discussed.

Animals↗

Chronic maxillary sinusitis associated with the mushroom Schizophyllum commune in a patient with AIDS.

Invasive infection with fungi of the Basidiomycota (rusts, smuts, toadstools, mushrooms, and puffballs) is extremely rare. We report such an infection in a patient with human immunodeficiency virus disease who presented with chronic maxillary sinusitis associated with the mushroom Schizophyllum commune. The organism was isolated from the surgical drainage material, and septate hyphae were seen invading the maxillary submucosa. The limited literature on this subject is reviewed.

Acquired Immunodeficiency Syndrome↗

Mushroom worker's lung resulting from indoor cultivation of Pleurotus osteatus.

Indoor cultivation of oyster mushroom Pleurotus osteatus lead to an outbreak of extrinsic allergic alveolitis in two workers. High titer of indirect fluorescent antibody and positive precipitins against basidiospores of P. osteatus were demonstrated in sera of the patients. Mushroom workers should protect themselves from the basidiospores, being aware of their pathogenicity.

Adult↗

Effects of mushroom and herb polysaccharides, as alternatives for an antibiotic, on the cecal microbial ecosystem in broiler chickens.

An in vivo experiment was conducted to study the potential prebiotic effects of mushroom and herb polysaccharide extracts, Lentinus edodes extract (LenE), Tremella fuciformis extract, and Astragalus membranaceus Radix extract, on chicken growth and the cecal microbial ecosystem, as compared with the antibiotic Apramycin (APR). This investigation was carried out in terms of a dose-response study. The chickens were naturally infected with avian Mycoplasma gallisepticum prior to the experiment. The BW gain, cecal pH, viscosity, and predominant microbial populations were measured 1 wk after the extract and APR treatments. The extracts and APR significantly stimulated growth of the chickens infected with avian Mycoplasma gallisepticum. The average BW gain of the groups fed with the extracts was significantly lower than that of the antibiotic group. The extracts had no significant effect on cecal pH. However, cecal viscosity and microbial populations were significantly affected by feeding extracts and antibiotic. In contrast to APR, the extracts stimulated the number of the potentially beneficial bacteria (bifidobacteria and lactobacilli), while reducing the number of the potentially harmful bacteria (Bacteroides spp. and Escherichia coli). Of the 3 extracts, LenE was associated with the most cecal bifidobacteria and lactobacilli. With each increase in the LenE dose, birds tended to have higher BW gain and total aerobe and anaerobe counts. Numbers of predominant cecal bacteria, in particular, E. coli, bifidobacteria, and lactobacilli, were significantly increased with increases in the LenE dose. It would seem that these specific mushroom and herb polysaccharide extracts hold some promise as potential modifiers of intestinal microbiota in diseased chickens.

Agaricales↗

Radiocaesium in mushrooms from northeast Italy, 1986-2002.

Late in the summer of 1986, the Health Physics Departments of Pordenone, Udine and Trieste, entrusted with monitoring radioactivity in the environment and food as a consequence of the Chernobyl accident, started noticing high concentrations of radionuclides--especially radiocaesium--in mushroom samples coming from different areas of the Friuli Venezia Giulia region (northeast Italy). To date, the authors have conducted 14 annual rounds of sampling and gamma spectrometry measurements on mushrooms, generating a total of over 2250 samples belonging to more than 300 species, which were picked in about 30 stations in the region. This surveys the main results from 15 years of macromycetes radiocontamination analysis in the region, the still unsolved problems, and hypotheses for future work.

Agaricales↗

Abnormal mushroom body plasticity in the Drosophila memory mutant amnesiac.

In Drosophila melanogaster, adult or larval rearing conditions influence brain structure. In particular, larval density affects the number of fibers forming the mushroom bodies, a neuropil structure involved in olfactory learning. The mushroom bodies receive chemosensory inputs from the antennal lobes at the level of the calyx. In this study we report that larval density affects calyx volume measured shortly after eclosion from the pupal case. We observe that in the memory mutant amnesiac this form of experience-dependent structural plasticity is missing, whereas it is not affected in the learning mutant rutabaga and in the memory mutant radish. Independent of the plasticity effect, the calyces are on average slightly bigger than wild type in amnesiac and smaller in rutabaga flies.

Amnesia↗

Hazard of a ruptured exhalation mushroom diaphragm.

In most breathing circuits, ventilation of a patient requires some device to occlude the expiratory limb during inflation. Usually one or another modification of a mushroom exhalation valve is employed. Such valves are found in the Bourns Bear 1 ventilator, the Bennett MA-1 and MA-2 ventilators, and the Emerson postoperative ventilator, as well as in the Frumen valve for manual ventilation. Recently, the authors observed a defect that might cause permanent occlusion, raising the patient's airway pressure to the level of the limiting safety pop-off and effectively applying a prolonged Valsalva maneuver without ventilation. The defect responsible for this malfunction was a half-centimeter laceration in the mushroom valve. Although the defect occurred on a Bourns Bear 1 ventilator, the authors were able to reproduce it on other ventilators of different manufacturers.

Cough↗

The evolution of non-reciprocal nuclear exchange in mushrooms as a consequence of genomic conflict.

Heterothallic mushrooms accomplish sex by exchanging nuclei without cytoplasm. Hyphal fusions occur between haploid mycelia resulting from germinated spores and subsequent reciprocal nuclear exchange without cytoplasmic mixing. The resulting dikaryon is therefore a cytoplasmic mosaic with uniformly distributed nuclei (two in each cell). Cytoplasmic inheritance is doubly uniparental: both mated monokaryons can potentially transmit their cytoplasm to the sexual spores, but normally only a single type per spore is found. Intracellular competition between mitochondria is thus limited, but at the dikaryon level, the two types of mitochondria compete over transmission. This creates the conditions for genomic conflict: within the dikaryon, a selfish mitochondrial mutant with increased relative transmission can be favoured, but selection between dikaryons will act against such a mitochondrial mutant. Moreover, because nuclear fitness is directly dependent on dikaryon fitness, a reduction in dikaryon fitness directly conflicts with nuclear interests. We propose that genomic conflict explains the frequent occurrence of non-reciprocal nuclear exchange in mushrooms. With non-reciprocal exchange, one monokaryon donates a nucleus and the other accepts it, but not vice versa as in the typical life cycle. We propose a model where non-reciprocal nuclear exchange is primarily driven by mitochondria inducing male sterility and the evolution of nuclear suppressors.

Agaricales↗

Pseudomonas costantinii sp. nov., another causal agent of brown blotch disease, isolated from cultivated mushroom sporophores in Finland.

Pathogenic bacteria are frequently associated with mushroom sporophores exhibiting brown blotch disease symptoms. These bacteria belong mainly to Pseudomonas tolaasii or occasionally to 'Pseudomonas reactans'. Although a group of isolates originating from some Finnish mushroom farms satisfied the two characteristic criteria for diagnosis of infection with P. tolaasii (i.e. yielding a typical brown blotch symptom on Agaricus bisporus sporophores and producing a typical white line in agar when streaked towards the 'P. reactans' LMG 5329 inducing strain), results based on numerical taxonomy, siderotyping, DNA-DNA hybridizations and 16S rDNA phylogenetic analyses supported the view that these isolates constituted a novel species within the genus Pseudomonas, Pseudomonas costantinii. The type strain is PS 3a(T) (= CFBP 5705(T) = HAMBI 2444(T)).

Agaricus↗

Improved molecular identification of Thermoactinomyces spp. associated with mushroom worker's lung by 16S rDNA sequence typing.

Mushroom worker's lung (MWL) is a hypersensitivity pneumonitis or allergic alveolitis caused by a type III IgG-mediated immunopathogenic inflammatory reaction in the host due to the inhalation of several thermophilic organisms, including Thermoactinomyces spp. It is difficult to distinguish phenotypically the eight species of this genus; therefore, this study sought to develop an improved molecular means of identifying Thermoactinomyces spp. associated with MWL by partial 16S rDNA PCR amplification and direct sequencing. Hypervariable regions within the 16S rRNA gene, which could be employed as signature sequences of the eight individual species, were identified and employed with highly conserved flanking primers to allow initial PCR amplification, before direct DNA sequencing of the 16S rDNA amplicons. A novel 24-mer 16S rDNA oligonucleotide upstream primer was designed from in silico alignments of all Thermoactinomyces spp. and was employed in combination with downstream (reverse) 16S rDNA primers. This permitted the successful identification of all four isolates associated with mushroom workers' lung. The method may be useful in the identification of Thermoactinomyces spp. associated with allergic alveolitis or pneumonitis associated with occupational exposure in agricultural and horticultural environments.

Agricultural Workers' Diseases↗