Microscopic evidence of sandstone deterioration and damage by fungi isolated from the Angkor monuments in simulation experiments.

The Angkor monuments have been registered on the World Cultural Heritage List of UNESCO, while the buildings built mostly of sandstone are suffering from serious deterioration and damage. Microorganisms are one of the leading causes for the sandstone deterioration. Identification of the mechanisms underlying the biodeterioration is of significance because it reveals the biochemical reaction involved so that effective conservation and restoration of cultural properties can be achieved. In this study, the fungal colonization and biodeterioration of sandstone in simulation experiments were examined using confocal reflection microscopy (CRM) and scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS). Aspergillus sp. strain AW1 and Paecilomyces sp. strain BY8 isolated from the deteriorated sandstone of Angkor Wat and Bayon of Angkor Thom, respectively, were inoculated and incubated with the sandstone used for construction of Angkor Wat. With CRM, we could visualize that strain AW1 tightly attached to and broke in the sandstone with extension of the hyphae. Quantitative imaging analyses showed that the sandstone surface roughness increased and the cavities formed under the fungal hyphae deepened during the incubation of strains AW1 and BY8. These highlighted that the massive growth of fungi even under the culture conditions was associated with the cavity formation of the sandstone and its expansion. Furthermore, SEM-EDS indicated the flat and Si-rich materials, presumably quartz and feldspar, were found frequently at the intact sandstone surface. But the flatness was lost during the incubation, possibly due to the detachment of the Si-rich mineral particles by the fungal deterioration. Consequently, this study proposed a biodeterioration model of the sandstone in that the hyphae of fungi elongated on the surface of the sandstone to penetrate into the soft and porous sandstone matrix, damaging the matrix and gradually destabilize the hard and Si-rich minerals, such as quartz and feldspar, to the collapse and cavities.

Bioactive sorbicillinoids from a rhizospheric soil-derived Paecilomyces sp. KMU21009.

A new hybrid sorbicillinoid named paeciureallin (1) and a new monomeric sorbicillinoid named paecillyketide (2), along with six known analogues (3-8), were isolated from the rhizospheric soil-derived fungus Paecilomyces sp. KMU21009 associated with Delphinium yunnanense. Their structures were elucidated by extensive spectroscopic analysis and comparison with literature values. Paeciureallin (1) is the first example of hybrid sorbicillinoids possessing a rare sorbicillinoid urea unit and containing a ß-D-ribofuranose functionality. In pharmacological studies, compounds 1 and 2 were evaluated for in vitro anti-inflammatory and cytotoxic activities. Paeciureallin (1) exhibited moderate cytotoxicity against SW480 and A549 cell lines, and the IC50 values were 32.0 ± 0.1 and 34.4 ± 2.0 µM, respectively.

Evaluation of antileishmanial activity of harzialactone a isolated from the marine-derived fungus Paecilomyces sp.

Fractionation of extracts from the culture broth of the marine-derived fungus, Paecilomyces sp. 7A22, resulted in the isolation of the harzialactone A (HA), a known compound previously isolated from fungi of marine environments. The chemical structure of HA was determined by spectroscopic analyses. Upon evaluation of HA on antileishmanial assays against Leishmania amazonensis, HA exhibited significant activity against promastigotes forms with IC50 of 5.25 µg mL-1 and moderate activity against intracellular amastigotes with IC50 of 18.18 µg mL-1. This is the first report on the antileishmanial activity of HA, and the effects of HA presented in this work suggest that this class of compounds are suitable for future biological in vitro and in vivo studies for the search of natural products with activity against Leishmania spp. Furthermore, the present results corroborate marine-derived fungi as a promising source of natural products with antiparasitic activity.

Trichosporon fermentans biomass flocculation from soybean oil refinery wastewater using bioflocculant produced from Paecilomyces sp. M2-1.

The soybean oil refinery (SOR) wastewater contains a high concentration of chemical oxygen demand (COD) and lipid, so the direct emissions of SOR wastewater will result in environmental pollution and waste of resources. Oleaginous yeast Trichosporon fermentans can consume organic materials in SOR wastewater to synthesize microbial oil, which achieves the purpose of SOR wastewater resource utilization. The effective harvesting technology of oleaginous yeasts can improve the utilization efficiency. In this study, Paecilomyces sp. M2-1 with high flocculating activity was isolated. The flocculants produced by M2-1 (MBF2-1) include 75% (w/w) polysaccharides, rely on cations, and display the flocculation percentage of above 77% in the range of pH 2-11. Especially under alkaline conditions, the flocculation percentage can be kept above 97%. The results of scanning electron microscope observation and zeta potential measurements suggested that the bridging, net trapping, and sweeping were the main flocculation mechanism of MBF2-1. MBF2-1 could flocculate T. fermentans that was used to reduce the organic matter in SOR wastewater and to produce microbial oil. Under the optimum conditions, the flocculation percentage of MBF2-1 against T. fermentans from SOR wastewater can reach 95%. Fatty acid content percent in microbial oil from T. fermentans was not almost affected by flocculation of MBF2-1. Moreover, MBF2-1 can further remove 55% and 53% of COD and oil content in the fermented SOR wastewater, respectively. The properties and high flocculating percentage displayed by MBF2-1 indicated its potential application prospect in oleaginous yeast harvest and food industry wastewater treatment.

Virulence evaluation of entomopathogenic fungi to subterranean termites, Globitermes sulphureus (INSECTA: ISOPTERA)

Aims: Subterranean termites, Globitermes sulphureus is one of the significant pests for agricultural crops such as coconut and oil palm, and occasionally attacks building structure in Malaysia. Efforts to control subterranean termite infestations depended heavily on liquid termiticide applications. Natural pathogen of termites such as entomopathogenic fungi (EPF) is a promising alternative to chemical control. The objective of this study was to determine the most virulent EPF such as Metarhizium anisopliae, Beauveria bassiana and Paecilomyces sp. against the subterranean termites, G. sulphureus for the development of bio-insecticide for future use of termite control. Methodology and results: Subterranean termites were collected from mound-building termites found in Universiti Malaysia Terengganu (UMT). Cultivating, harvesting and counting of conidia of EPF were carried out to prepare the desired concentrations for screening test and bioassays, which were 1×103 , 1x105 , 1×107 and control. The pathogenicity testing was observed daily within 1 week under laboratory conditions. Screening showed that M. anisopliae was found to be the most virulent compared to B. bassiana and Paecilomyces sp., achieving 100% mortality within 3-4 days. M. anisopliae was then further tested on termites and it was found that concentration of 1×107 showed the lowest LT50 value, while LC50 of M. anisopliae in 1 day was 2.0151×106 . Conclusion, significance and impact of study: It could be concluded that M. anisopliae is the most virulent EPF against termites and the most effective concentration was ±107 followed by strains of B. bassiana and Paecilomyces sp. However, there is limited field evaluation of EPF against termites in Malaysia. Evaluation on the efficiency of M. anisopliae in the field should be conducted so that its efficacy could be proven and marketed.

Composting of food waste subjected to hydrothermal pretreatment and inoculated with Paecilomyces sp. FA13.

Food waste collected from restaurants, convenience stores, and food-processing factories was mixed with sawdust and subjected to hydrothermal pretreatment at 180°C for 30min to prepare compost raw material. Furan compounds such as 5-HMF (5-hydroxymethyl furfural) and furfural were produced at concentration levels of approximately 8 and 0.5mg/g-ds, respectively, through hydrothermal pretreatment. The furan compounds inhibited the activity of composting microorganisms, thus delaying the start of organic matter degradation during composting. A newly identified fungus, Paecilomyces sp. FA13, which possesses the ability to degrade furan compounds, was isolated and used as an inoculum for the composting of the raw material prepared by hydrothermal pretreatment. By inoculating the FA13 into the compost raw material at 10(5)CFU/g-ds, the degradation of furan compounds was accelerated. As a result, bacterial activity, which contributed to composting, was enhanced, significantly promoting the start of vigorous degradation of organic materials.

Absolute configurations of four resorcylic acid lactones, paecilomycins J-M, by CD/TDDFT calculations.

The absolute configurations of four resorcylic acid lactones (RALs), paecilomycins J-M (1-3 and 5), were assigned by Time-Dependent Density-Functional Theory (TDDFT) calculations of their electronic circular dichroism (CD) spectra. The previously reported structure 4 for paecilomycin M was found to be incorrect and should be changed to structure 5. Analysis of structure-spectrum relationship for this group of RALs suggested that V'-shape conformations give type I CD spectra (two negative Cotton effects around 300 and 260 nm, a positive Cotton effect around 220 nm) while V-shape conformations yield type II spectra (signs of three Cotton effects were opposite to those in type I).

Variability in the production of extracellular enzymes by entomopathogenic fungi grown on different substrates

Entomopathogenic fungi are important controllers of pest-insects populations in agricultural production systems and in natural environment. These fungi have enzymatic machinery which involve since the recognition and adherence of spores in their hosts culminating with infection and death of these insects. The main objective of this study was to analyzed extracellular enzyme production of the fungi strains Beauveria bassiana, Metarhizium anisopliae and Paecilomyces sp when cultured on substrates. These fungi were grown in minimal media containing specific substrates for the analysis of different enzymes such as amylases, cellulases, esterases, lipases, proteases (gelatin and caseinase), pectinases and cuticles of Musca domestica larvae and adults. All the assays were performed with and without the presence of dextrose in the culture media. The quantification of enzyme activity was performed by the ratio of halo / colony (H/C) and the results subjected to variance analysis level of 5% (ANOVA) followed by post-Tukey test. All strains were positive for lipase and also they showed a high significant enzyme production for gelatin at concentrations of 4 and 1%. B. bassiana and Paecilomyces sp. were positive for amylase, pectinase and caseinase, and only Paecilomyces sp. showed cellulase activity.

Variability in the production of extracellular enzymes by entomopathogenic fungi grown on different substrates.

Entomopathogenic fungi are important controllers of pest-insects populations in agricultural production systems and in natural environment. These fungi have enzymatic machinery which involve since the recognition and adherence of spores in their hosts culminating with infection and death of these insects. The main objective of this study was to analyzed extracellular enzyme production of the fungi strains Beauveria bassiana, Metarhizium anisopliae and Paecilomyces sp when cultured on substrates. These fungi were grown in minimal media containing specific substrates for the analysis of different enzymes such as amylases, cellulases, esterases, lipases, proteases (gelatin and caseinase), pectinases and cuticles of Musca domestica larvae and adults. All the assays were performed with and without the presence of dextrose in the culture media. The quantification of enzyme activity was performed by the ratio of halo / colony (H/C) and the results subjected to variance analysis level of 5% (ANOVA) followed by post-Tukey test. All strains were positive for lipase and also they showed a high significant enzyme production for gelatin at concentrations of 4 and 1%. B. bassiana and Paecilomyces sp. were positive for amylase, pectinase and caseinase, and only Paecilomyces sp. showed cellulase activity.

Compatibility of entomopathogenic fungi with neonicotinoid insecticides / Compatibilidade de fungos entomopatogênicos com inseticidas nicotinóides

O efeito fungitóxico, in vitro, de três inseticidas neonicotinóides, acetamiprid (Saurus 200 PS), imidacloprid (Confidor 700 GrDA) e thiamethoxam (Actara 250 WG) em três concentrações (RM = recomendação média para campo, 0,7 RM e 1,3 RM), foi avaliado sobre os fungos entomopatogênicos Beauveria bassiana, Metarhizium anisopliae e Paecilomyces sp. O efeito dos inseticidas sobre a germinação dos conídios, o crescimento vegetativo e a produção de conídios também foi avaliado. Os resultados mostraram que os produtos não afetaram a germinação dos conídios, com exceção do acetamiprid na maior concentração (1,3 RM) onde ocorreu uma inibição (Pú0,05) de M. anisopliae. O crescimento vegetativo foi inibido nos tratamentos acetamiprid, nas três concentrações, para B. bassiana e M. anisopliae. Também no tratamento thiamethoxam, na concentração RM e 1,3 RM o crescimento vegetativo foi inferior à testemunha para B. bassiana. Para Paecilomyces sp. o crescimento vegetativo foi superior ao da testemunha para os tratamentos 0,7 RM e RM de acetamiprid, todas as concentrações de imidacloprid e as duas menores de thiamethoxam. A produção de conídios foi menor para o tratamento de acetamiprid na maior concentração (1,3 RM) para os fungos Paecilomyces sp. e M. anisopliae e para imidacloprid na maior concentração (1,3 RM) para Paecilomyces sp. Já thiamethoxam inibiu a produção de conídios somente na menor concentração (0,7 RM) para M. anisopliae. Aumento de produção de conídios foi observado somente nos tratamentos acetamiprid nas duas menores concentrações (0,7 RM) e RM e thiamethoxam nas duas maiores (RM e 1,3 RM) para o fungo Paecilomyces sp. Assim, os inseticidas testados nas concentrações e formulações utilizadas mostraram, na maioria dos casos, compatibilidade com os entomopatógenos B. bassiana, M. anisopliae e Paecilomyces sp. Portanto, estes produtos poderão ser recomendados em programas de MIP, nas formulações e concentrações testadas, para o controle...

The in vitro fungitoxic effect of the neonicotinoid insecticides acetamiprid (Saurus 200 SP), imidacloprid (Confidor 700 WDGr) and thiamethoxam (Actara 250 WG) in three concentrations (AR= average field recommendation; 0.7 AR and 1.3 AR) to the entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae and Paecilomyces sp. was studied. The effect of the insecticides on conidia germination, vegetative growth and conidiogenesis was compared. The insecticides had no effect on conidia germination, except under the highest concentration (1.3 AR) of acetamiprid, in which significant inhibition of M. anisopliae occurred. Vegetative growth of B. bassiana and M. anisopliae was significantly inhibited only by the three concentrations of acetamiprid treatment. Thiamethoxam treatment, in the AR and 1.3 AR concentrations, was significantly lower for B. bassiana vegetative growth. Paecilomyces sp. vegetative growth was higher than the control in the following treatments: 0.7 AR and AR of acetamiprid; all concentrations of imidacloprid; and in the two smallest concentrations, 0.7 AR and AR, of thiamethoxam. Conidia production was significantly smaller for Paecilomyces sp. and M. anisopliae, in the highest concentration (1.3 AR) of acetamiprid treatment and, for Paecilomyces sp., in the highest concentration (1.3 AR) of imidacloprid. Thiamethoxam significantly inhibited M. anisopliae conidia production, only in the smallest concentration (0.7 AR).Significant increase in the conidia production of Paecilomyces sp. fungus was observed in the following treatments: two concentrations, 0.7 AR and AR of acetamiprid; and in the two highest concentrations, AR and 1.3 AR, of thiamethoxam treatments. The use of insecticides, in the recommended formulations and other concentrations tested, in most cases, had no negative effect on conidia germination, conidia production and vegetative growth of B. bassiana, M. anisopliae and Paecilomyces sp. Consequently...