Antioxidant Activity and Infrared Spectroscopy Analysis of Alcoholic Extracts Obtained from Paecilomyces hepiali (Ascomycetes).

This research studies the influence of substrate on the antioxidant activity of alcohol extracts of Paecilomyces hepiali. We used corn, rice, millet, and peas as substrates. Antioxidant activity was measured with the DPPH radical scavenging method. Concentrations of extracts (6.25, 3.12, 1.56, 0.78, and 0.39 mg/mL) were applied in all evaluations. Overall antioxidant activity was expressed as the concentration of substrate that decreased DPPH radical levels by 50% (IC50DPPH) for 7 methanol and 7 ethanol extracts. A comparison of IC50DPPH allowed us to conclude that the methanol extracts are more active in scavenging stable DPPH radicals than are the ethanol extracts. The substrate with antioxidant properties most suitable for cultivation of P. hepiali was rice supplemented with non-defatted soy flour. The extract most effective in scavenging stable radicals was the methanol extract of sample 4 (IC50DPPH = 2.33 mg/mL) cultivated on rice with nondefatted soy flour. The methanol extract of sample 7 cultivated on peas was less effective (IC50DPPH = 11.50 mg/mL). By crystallizing these extracts, we managed to obtain sufficient quantities of 6 samples in a solid state, for which infrared spectra were measured and confirmed the presence of amino acids in the extracts.

Transcriptome analysis of Paecilomyces hepiali at different growth stages and culture additives to reveal putative genes in cordycepin biosynthesis.

Paecilomyces hepialid (PH) is an endoparasitic fungus of Cordyceps sinensis (CS) and has become a substitute for CS due to their similar pharmacological activities. Because the market demand for CS is difficult to satisfy, and cordycepin, the effective compound of CS, is difficult to industrially produce, we produced 5 samples of PH by culturing for different durations and adding different additives to the culture broth, and detected their cordycepin content with UPLC ESI MS/MS. Then we grouped these cultures into five transcriptome comparisons containing 3 time variable groups and 2 additive variable groups. We used next-generation (NG) sequencing methods to acquire transcriptomic information and investigated the response of gene expression to the additives and the influence of different growth stages. This work will contribute to a better understanding of purine metabolism in PH, and possibly in other Cordyceps species. It will provide a useful resource to further advance transcriptomics studies in Cordyceps species.

Total Phenolic Content and Antioxidant Activity of Mycelial Extracts from the Medicinal Fungus Paecilomyces hepiali (Ascomycetes).

Paecilomyces hepialid, a fungus originating in the Tibetan Plateau, has been used as a substitute for Ophiocordyceps sinensis because the primary chemical compounds and pharmacological effects of P. hepialid are similar to those of O. sinensis. P. hepialid has been developed into a dietary supplement and pharmaceutical products. The antioxidant activity of extracts using 2 solvents (water and ethanol) from mycelia obtained from 2 cultivation modes (solid-state and submerged cultivation) were evaluated in this study. Four strains of P. hepialid obtained from Qinghai, Sichuan, and Yunnan Provinces were included; the total phenolic content and in vitro antioxidant activity of mycelial extracts were compared. The total phenolic content and antioxidant activity of strains were found to be affected by the cultivation mode and extraction solvent. The ethanol extracts of solid-state cultivation of strain 2138, obtained from Sichuan Province, exhibited the highest antioxidant activity. The results showed that different strains might require different cultivation modes and extraction solvents for better antioxidant activity. However, solid-state cultivation and ethanol extraction are generally recommended based on the analyses conducted. Strain 2138 may be a good candidate for the purpose of producing functional foods. The results suggest that strain selection is important when P. hepialid is used to manufacture pharmaceutical products.

White mold on cultivated morels caused by Paecilomyces penicillatus.

Morchella (morel) includes prized edible and medical mushrooms in the world. Since 2012, commercial cultivation of morels in the field has developed rapidly in China. However, coupled with the rapid expansion of morel cultivation, diseases have been become serious threats to morel production. White mold is one of the most serious diseases on cultivated morels. This study aimed to confirm this pathogen by following Koch's postulates, and to identify it using molecular evidence. Our results indicated that healthy Morchella fruiting bodies inoculated with Paecilomyces sp. isolates produced typical white mold symptoms, and the internal transcribed spacer sequences of the Paecilomyces sp. were 99% similar to that recovered from an epitype of Paecilomyces penicillatus. Therefore, P. penicillatus was considered to be the causative agent of white mold. White mold occurred from the initial harvest to the storage and preservation process, and it produced white mold-like symptoms on the caps and stripes of Morchella. This is the first time that white mold has been reported on cultivated Morchella.

Antiparasitic drugs: in vitro tests against nematophagous fungi / Antiparasitários: teste in vitro contra fungos nematófagos

Abstract The use of biological agents has been intensified in recent years against eggs and larvae of gastrointestinal nematodes as an alternative control method in pasture plant health management, with the concomitant use with antiparasitic drugs still occurring. The aim of this study was to test the in vitro activity of the following antiparasitic drugs: Ivermectin and albendazole against the following nematophagous fungi: Paecilomyces fumosoroseus, Paecilomyces lilacinus and Paecilomyces variotii. The agar diffusion test was performed using an initial concentration of 0.0016g/mL of each drug, after solidification of the culture medium containing the drug concentration each nematophagous fungi was inoculated. The results showed that in a concentration of 80μg/mL, the fungal growth decreased, however, with the concentration of 160μg/mL, there was no fungal growth in both drugs, compared to the control, which indicates an inhibition in the development of the nematophagous fungi studied when they come in contact with ivermectin and albendazole.

Resumo O uso de agentes biológicos que atuam em ovos e larvas de nematódeos gastrintestinais como uma alternativa para o manejo de pastagens de saúde tem se intensificado nos últimos anos, bem como o uso concomitante com outros medicamentos antiparasitários. O objetivo deste estudo foi testar o efeito in vitro dos fármacos Ivermectina e Albendazol em fungos nematófagos Paecilomyces fumosoroseus, Paecilomyces lilacinus e Paecilomyces variotii. Foi utilizada a técnica de difusão em agar, sendo preparado a partir de uma concentração inicial de 0,0016g/mL de cada uma das drogas e diluídas em meio de cultura, com posterior semeadura dos fungos nematófagos. Os resultados mostraram que na concentração de 80μg/mL, o crescimento diminuiu, no entanto, com a concentração de 160μg/mL de ambas as drogas, não houve crescimento de fungos durante o período de estudo, em comparação com o controle, indicando a inibição do desenvolvimento dos fungos nematófagos estudados quando em contato com a Ivermectina e Albendazol.

Antiparasitic drugs: in vitro tests against nematophagous fungi.

The use of biological agents has been intensified in recent years against eggs and larvae of gastrointestinal nematodes as an alternative control method in pasture plant health management, with the concomitant use with antiparasitic drugs still occurring. The aim of this study was to test the in vitro activity of the following antiparasitic drugs: Ivermectin and albendazole against the following nematophagous fungi: Paecilomyces fumosoroseus, Paecilomyces lilacinus and Paecilomyces variotii. The agar diffusion test was performed using an initial concentration of 0.0016g/mL of each drug, after solidification of the culture medium containing the drug concentration each nematophagous fungi was inoculated. The results showed that in a concentration of 80µg/mL, the fungal growth decreased, however, with the concentration of 160µg/mL, there was no fungal growth in both drugs, compared to the control, which indicates an inhibition in the development of the nematophagous fungi studied when they come in contact with ivermectin and albendazole.

Potential biodegradation of crude petroleum oil by newly isolated halotolerant microbial strains from polluted Red Sea area.

Two microbial isolates from oil polluted Red Sea water in Egypt, designated as RS-Y1 and RS-F3, were found capable of degrading Belayim mix (BX) crude oil. Strains RS-Y1 and RS-F3 were assigned to the genera Lipomyces tetrasporus and Paecilomyces variotii based on their morphological and physiological characteristics. Both isolates were compared for the biodegradation of crude petroleum-oil hydrocarbons in basal salt medium supplemented with 5% (w/v) of BX-crude oil. Gas chromatography profile showed that the biodegradation of total petroleum hydrocarbons (TPHs) inoculated with L. tetrasporus (68.3%) and P. variotii (58.15%) along with their consortium (66%) significantly reduced TPHs levels as compared to the control after 30days. L. tetrasporus (44.5%) was more effective than P. variotii strain (32.89%) in reducing the unresolved complex mixtures (UCM) content from the medium. Both isolates exhibited a strong growth over a wide range of salinity (5-45g/L NaCl).

Growth of Paecilomyces variotii in B0 (diesel), B100 (biodiesel) and B7 (blend), degradation and molecular detection.

The introduction of biodiesel to diesel may allow the fuel to be more susceptible to microorganism growth, especially during incorrect storage. To evaluate the effect of adding biodiesel in pure diesel on the growth of Paecilomyces variotii, microcosms containing pure diesel (B0), blend diesel/biodiesel (B7) and pure biodiesel (B100) were used. In microcosm with minimal mineral medium and B0, B7 or B100, after 60 days, the biomass (dry weight) formed at interface oil-water in B7 and B100 was significantly higher when compared to that of B0. Infrared analysis showed reduction of the carbonile fraction in B7 and B100 suggesting formation of intermediate compounds in B7. To monitor possible contamination of fuel storage tank by P. variotii samples were collected and analysed by specific-PCR assay for detection of P. variotii spores in the aqueous phase. This method was able to detect a minimum of 103 spores ml-1, corresponding to 0.0144 ng µl-1 of DNA. Specificity was tested against Aspergillus fumigatus and Pseudallescheria boydii.

Growth of Paecilomyces variotii in B0 (diesel), B100 (biodiesel) and B7 (blend), degradation and molecular detection / Crescimento de Paecilomyces variotii em B0 (diesel), B100 (biodiesel) e B7 (mistura), degradação e detecção molecular

AbstractThe introduction of biodiesel to diesel may allow the fuel to be more susceptible to microorganism growth, especially during incorrect storage. To evaluate the effect of adding biodiesel in pure diesel on the growth of Paecilomyces variotii, microcosms containing pure diesel (B0), blend diesel/biodiesel (B7) and pure biodiesel (B100) were used. In microcosm with minimal mineral medium and B0, B7 or B100, after 60 days, the biomass (dry weight) formed at interface oil-water in B7 and B100 was significantly higher when compared to that of B0. Infrared analysis showed reduction of the carbonile fraction in B7 and B100 suggesting formation of intermediate compounds in B7. To monitor possible contamination of fuel storage tank by P. variotii samples were collected and analysed by specific-PCR assay for detection of P. variotii spores in the aqueous phase. This method was able to detect a minimum of 103 spores ml–1, corresponding to 0.0144 ng µl–1 of DNA. Specificity was tested against Aspergillus fumigatus and Pseudallescheria boydii.

ResumoA introdução de biodiesel ao diesel pode permitir que o combustível se torne mais suscetível ao crescimento de microorganismos, especialmente durante o armazenamento incorreto. Para analisar o efeito da adição de biodiesel em diesel puro no crescimento de Paecilomyces variotii, avaliou-se seu desenvolvimento em microcosmos contendo diesel puro (B0), mistura diesel/biodiesel (B7) e biodiesel puro (B100). Em microcosmos com meio mineral mínimo e B0, B7 ou B100, após 60 dias, a biomassa (peso seco) formada na interface óleo-agua com B7 e B100 foi significativamente maior quando comparada com a de B0. A análise de infravermelho mostrou redução da fração carbonila em B7 e B100, sugerindo a formação de compostos intermediários em B7. Para monitorar uma possível contaminação de tanque de armazenamento de combustível por P. variotii, amostras foram colhidas e analisadas por um teste de PCR específico para detecção de esporos deste fungo em fase aquosa. Este método foi capaz de detectar um mínimo de 103 esporos ml–1, correspondente a 0.0144 ng µl–1 de DNA. Especificidade foi testada contra Aspergillus fumigatus e Pseudallescheria boydii.

Conversion of food waste into biofertilizer for the biocontrol of root knot nematode by Paecilomyces lilacinus.

The feasibility of converting food waste into nematocidal biofertilizer by nematophagous fungus Paecilomyces lilacinus (P. lilacinus) was investigated. The culture conditions of P. lilacinus were optimized through response surface methodology. Results showed that fermentation time, the amount of food waste, initial pH and temperature were most important factors for P. lilacinus production. The P. lilacinus production under optimized conditions was 10(9.6 ± 0.3) conidia mL⁻¹. After fermentation, the chemical oxygen demand concentration of food waste was efficiently decreased by 81.92%. Moreover, the property evaluation of the resultant food waste as biofertilizer indicates its high quality with reference to the standard released by the Chinese Ministry of Agriculture. The protease activity and nematocidal ability of P. lilacinus cultured by food waste were 10.8% and 27% higher than those by potato dextrose agar, respectively.

Nematicidal activity of Paecilomyces lilacinus 6029 cultured on Karanja cake medium.

Antagonistic fungi parasitize root-knot nematodes through secretion of extracellular hydrolytic enzymes and secondary metabolites. In present study, in vitro bioassay showed that Paecilomyces lilacinus 6029 culture filtrate from Karanja cake medium killed 100% Meloidogyne incognita larvae while only 78.28% mortality was recorded by Czapeck-Dox filtrate within 12 h of exposure. The filtrate, irrespective of culture medium, was found to be more nematotoxic when incubated for 15 days. Fourier Transform infrared spectroscopy predicted the presence of phenolic and alcoholic compounds in the filtrate. Furthermore, the active metabolites in fungal filtrate were partially characterized. pH stability test revealed that nematotoxicity of the filtrate appeared at all range of pH with low pH filtrate possessing more toxicity against M. incognita. Interestingly, buffers of same pH value did not show any nematicidal effect. No significant difference in nematicidal activity was observed between boiled (98.2% mortality) and unboiled culture filtrate (100% mortality). Ethyl acetate and lyophilized aqueous extracts produced higher nematicidal activity than a hexane extract indicating polar nature of active compounds produced by P. lilacinus 6029.

The entomopathogenic fungal endophytes Purpureocillium lilacinum (formerly Paecilomyces lilacinus) and Beauveria bassiana negatively affect cotton aphid reproduction under both greenhouse and field conditions.

The effects of two entomopathogenic fungal endophytes, Beauveria bassiana and Purpureocillium lilacinum (formerly Paecilomyces lilacinus), were assessed on the reproduction of cotton aphid, Aphis gossypii Glover (Homoptera:Aphididae), through in planta feeding trials. In replicate greenhouse and field trials, cotton plants (Gossypium hirsutum) were inoculated as seed treatments with two concentrations of B. bassiana or P. lilacinum conidia. Positive colonization of cotton by the endophytes was confirmed through potato dextrose agar (PDA) media plating and PCR analysis. Inoculation and colonization of cotton by either B. bassiana or P. lilacinum negatively affected aphid reproduction over periods of seven and 14 days in a series of greenhouse trials. Field trials were conducted in the summers of 2012 and 2013 in which cotton plants inoculated as seed treatments with B. bassiana and P. lilacinum were exposed to cotton aphids for 14 days. There was a significant overall effect of endophyte treatment on the number of cotton aphids per plant. Plants inoculated with B. bassiana had significantly lower numbers of aphids across both years. The number of aphids on plants inoculated with P. lilacinum exhibited a similar, but non-significant, reduction in numbers relative to control plants. We also tested the pathogenicity of both P. lilacinum and B. bassiana strains used in the experiments against cotton aphids in a survival experiment where 60% and 57% of treated aphids, respectively, died from infection over seven days versus 10% mortality among control insects. Our results demonstrate (i) the successful establishment of P. lilacinum and B. bassiana as endophytes in cotton via seed inoculation, (ii) subsequent negative effects of the presence of both target endophytes on cotton aphid reproduction using whole plant assays, and (iii) that the P. lilacinum strain used is both endophytic and pathogenic to cotton aphids. Our results illustrate the potential of using these endophytes for the biological control of aphids and other herbivores under greenhouse and field conditions.

Evaluation of Paecilomyces variotii potential in bioethanol production from lignocellulose through consolidated bioprocessing.

The ascomycete Paecillomyces variotii was evaluated for the first time as a candidate species for the production of bioethanol from lignocellulose through consolidated bioprocessing (CBP) approaches. The examined strain (ATHUM 8891) revealed all the necessary phenotypic characteristics required for 2nd generation biofuel production. The fungus is able to efficiently ferment glucose and xylose to ethanol, with yields close to the theoretical maximum. Nitrogen supplementation greatly affected ethanol production with nitrate-nitrogen presenting the best results. Notably, ethanol yield on xylose fermentation was higher than that of glucose, while in co-fermentation of glucose-xylose mixtures no distinguished diauxic behavior was observed. Furthermore, the fungus seems to possess the necessary enzyme factory for the degradation of lignocellulosic biomass, as it was able to grow and produce ethanol on common agro-industrial derivatives. Overall, the results of our study indicate that P. variotii is a new and possibly powerful candidate for CBP applications.

Is there a role for statins in fungal infections?

It has been hypothesized that statins, HMG-CoA reductase inhibitors, may be used to treat fungal infections. Here we review data on antifungal properties of statins, effects on the host inflammatory response as well as available clinical evidence. We conclude that: statins exhibit antifungal properties in vitro although at supraphysiological concentrations; statins appear to have anti-inflammatory effects on host cells in vitro; statins have effects on fungal physiology beyond direct growth inhibition; clinical studies are scarce (n = 5), and their design is retrospective and observational, which is associated with a high risk of bias. Given the limited evidence for a beneficial effect of statins in fungal infection, randomized and controlled trials are highly warranted in this field.

[The effect of low radiation doses on the growth of Aspergillus versicolor and Paecilomyces lilacinus].

Peculiarities of growth of the strains of two species of microscopic fungi Aspergillus versicolor and Paecilomyces lilacinus were analyzed under exposure to chronic irradiation. It was shown that the rate of radial growth increased nonuniformly in the range of doses from 0 to 250 mGy but with maximums at a certain dose of radiation. It was shown that the absorbed dose of radiation to 2 Gy did not influence the survival of investigated strains in comparison with the control ones without irradiation. It was shown that the doses of irradiation to 2 Gy (at capacity exposure dose 0.955 microC/kg) are small for these species of microscopic fungi.

Alachlor oxidation by the filamentous fungus Paecilomyces marquandii.

Alachlor, a popular chloroacetanilide herbicide, can be a potential health risk factor. Soil microorganisms are primarily responsible for conversion and migration of alachlor in natural environment, but knowledge concerning alachlor biodegradation is not complete. Therefore, we studied the ability of Paecilomyces marquandii, soil fungus tolerant to heavy metals, to eliminate alachlor and proposed a new pathway of its transformation. After 7 days of incubation only 3.3% of alachlor was detected from an initial concentration 50 mg L(-1) and 20.1% from a concentration 100 mg L(-1). The qualitative IDA LC-MS analysis showed the presence of ten metabolites. All of them were dechlorinated mainly through oxidation, but also reductive dechlorination was observed. The main route of alachlor conversion progressed via N-acetyl oxidation resulting in the formation of mono-, di- and trihydroxylated byproducts. N-acetyl oxidation as a dominant route of alachlor metabolism by fungi has not been described so far. The toxicity of alachlor tested with Artemia franciscana did not increase after treatment with P. marquandii cultures. Paecilomyces marquandii strain seems to be an interesting model for the research on alachlor conversion by soil microscopic fungi, due to its dechlorination and hydroxylation ability as well as high tolerance to heavy metals.

Culture conditions and medium components for the production of mycelial biomass and exo-polysaccharides with Paecilomyces japonica in liquid culture.

In this study, the liquid culture conditions were optimized for maximal production of mycelial biomass and exo-polysaccharide by Paecilomyces japonica. The effects of medium composition, C/N ratio and physical parameters were investigated. From these experiments, 30 g glucose, 20 g yeast extract, 0.5 g KH2PO4, and 0.1 g CuCl2 2H2O in 1-l distilled water were found to be the most suitable carbon, nitrogen, and mineral sources, respectively. The optimal temperature, initial pH, agitation, and aeration were determined to be 27°C, uncontrolled pH, 400 rpm, and 1.0 vvm, respectively. Under these optimal conditions, the maximum mycelial growth and polysaccharides production were 23.1 g/l and 2.5 g/l, respectively.

Abundance and dynamics of filamentous fungi in the complex ambrosia gardens of the primitively eusocial beetle Xyleborinus saxesenii Ratzeburg (Coleoptera: Curculionidae, Scolytinae).

Insect fungus gardens consist of a community of interacting microorganisms that can have either beneficial or detrimental effects to the farmers. In contrast to fungus-farming ants and termites, the fungal communities of ambrosia beetles and the effects of particular fungal species on the farmers are largely unknown. Here, we used a laboratory rearing technique for studying the filamentous fungal garden community of the ambrosia beetle, Xyleborinus saxesenii, which cultivates fungi in tunnels excavated within dead trees. Raffaelea sulfurea and Fusicolla acetilerea were transmitted in spore-carrying organs by gallery founding females and established first in new gardens. Raffaelea sulfurea had positive effects on egg-laying and larval numbers. Over time, four other fungal species emerged in the gardens. Prevalence of one of them, Paecilomyces variotii, correlated negatively with larval numbers and can be harmful to adults by forming biofilms on their bodies. It also comprised the main portion of garden material removed from galleries by adults. Our data suggest that two mutualistic, several commensalistic and one to two pathogenic filamentous fungi are associated with X. saxesenii. Fungal diversity in gardens of ambrosia beetles appears to be much lower than that in gardens of fungus-culturing ants, which seems to result from essential differences in substrates and behaviours.

Optimization of fermentation process for the production of intracellular polysaccharide from Paecilomyces cicadae and the immuno-stimulating activity of intracellular polysaccharide.

Optimization of fermentation process for the production of intracellular polysaccharide (IPS) from the fungus Paecilomyces cicadae and the immuno-stimulating activity of IPS were carried out. The quantitative effects of initial pH, fermentation temperature and time on the yield of IPS content produced by P. cicadae in submerged fermentation were investigated separately using response surface methodology (RSM). The three factors chosen for the present investigation were based on the results of a previous Plackett-Burman (PB) design. The experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis and also analyzed by appropriate statistical methods. RSM analysis showed good correspondence between experimental and predicted values. It was found that three parameters represented significant effect. Probability value (p < 0.0001) demonstrated a very high significance for the regression model. By solving the regression equation and analyzing the response surface contour plots, the optimal process parameters were determined, i.e. fermentation temperature 24.53 °C, initial pH 7.46 and fermentation time 73.9 h. The maximum predicted yield of IPS was 356.02 µg/ml under the optimal conditions. Meanwhile, IPS from P. cicadae was found to have direct immuno-stimulating activity in vitro on murine macrophage RAW264.7 proliferative response and to stimulate nitric oxide generation in a dose-dependent manner.

Protein increase and lysine production by a Paecilomyces variotii strain grown on two-phase olive mill waste.

Two-phase olive-mill waste, the so-called "ecological", has been treated with a Paecilomyces variotii isolate in solid state fermentation experiments. The growth of the microorganism was estimated by measuring the production of carbon dioxide, using gas chromatography. A 46% increase of the protein content was achieved at the fermented product, after molasses addition at the initial mixture. The amino acid profile of the produced protein, as far as the essential amino acids are concerned, was significantly improved, resulting in a product that has the potential to be used as animal feed. Furthermore, it contains lysine, one of the essential amino acids that did not exist at the original product and is produced during fermentation. This is the first report on solid state fermentation of the two-phase olive mill waste (TPOMW) as a substrate, using a Paecilomyces variotii strain.