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Due to the increasing populations of anthelmintic-resistant gastrointestinal nematodes and as a consequence of the adverse effects of synthetic drugs, this study focuses on the search for secondary metabolites with nematocidal activity from the edible mushroom Pleurotus djamor using The proton nuclear magnetic resonance (1H-NMR) metabolomics. The highest activity was shown by the ethyl acetate fractions of mycelium (EC50 290.8 µg/mL) and basidiomes (EC50 282.7 µg/mL). Principal component analysis (PCA) and hierarchical data analysis (HCA) of the 1H-NMR metabolic profiles data showed that the ethanolic extracts, the ethyl acetate, butanol, and water fractions from mycelium have different metabolic profiles than those from basidiomes, while low polarity (hexane) fractions from both stages of fungal development show similar profiles. Orthogonal partial least squares discriminant analysis (OPLS-DA) allowed the identification of signals in the 1H-NMR metabolic profile associated with nematocidal activity. The signals yielded via OPLS-DA and bidimensional NMR analysis allowed the identification of uracil as a component in the ethyl acetate fraction from basidiomes, with an EC50 of 237.7 µg/mL. The results obtained showed that chemometric analyses of the 1H-NMR metabolic profiles represent a viable strategy for the identification of bioactive compounds from samples with complex chemical profiles.
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To explore the capability of soil mycobiota to degrade avocado peel waste and identify relevant successions and trophic guild shifts, fungal communities from three environments with different land uses were evaluated in a solid-state process. Soil samples used as inoculum were collected from a pristine mature tropical forest, a traditionally managed Mayan land, and an intensively managed monospecific avocado plantation. Soil-substrate mixes were evaluated for 52 weeks to evaluate organic matter decay and the carbon-to-nitrogen ratio. Amplicon-based high-throughput sequencing from internally transcribed spacer (ITS) analysis revealed significant differences in fungal communities widely dominated by Fusarium sp. and Clonostachys sp.; however, less represented taxa showed relevant shifts concomitantly with organic matter content drops. Trophic guild assignment revealed different behaviors in fungal communities between treatments over the 52 weeks, suggesting distinct preconditioning of fungal communities in these environments. Overall, the results lead to the identification of promising degradation moments and inoculum sources for further consortia enrichment or bioprospecting efforts.
Assuntos
Micobioma , Persea , Florestas , Fungos , Microbiologia do SoloRESUMO
The objective of this study was to evaluate and compare the in vitro lethal effect of the hydroalcoholic extract of the spent substrate of Pleurotus djamor ECS-123, obtained at 15 days of colonization (SPS) and at the first (SPS1) and second (SPS2) harvests, against infective larvae L3 of Haemonchus contortus. The in vitro lethal effect was evaluated by the L3 larval mortality test (LM) using six concentrations: 1.25, 2.5, 5, 10, 20, and 40 mg/mL, with ivermectin and thiabendazole (5 mg/mL) as controls. The first harvest extract (SPS1) of strain ECS-123 was subjected to liquid-liquid bipartition, which resulted in two fractions: aqueous (PdAcO) and ethyl acetate (PdAct). The chemical fractionation of PdAct with the highest mortality rate (80.11 %) was carried out with open-column chromatography, giving a total of 13 fractions, which were analyzed by thin-layer chromatography (TLC) and grouped into 5 mixtures (R1;1-3, R2;4-7, R3;8-9, R4;10-11 and R5;12-13). Subsequently, the mixtures were evaluated against H. contortus L3 larvae. Finally, the components of the mixtures with the highest nematocidal effects were evaluated by gas chromatography coupled to mass spectrometry (GC-MS). The data were analyzed with a completely randomized design through ANOVA using the generalized linear model (GLM) with the "R" program. The purification and characterization of R4 and R5 by GC-MS revealed the presence of the following compounds: veratryl alcohol, 4-hydroxy-3,5,5 trimethyl-4-[3-oxo-1-butenyl]-2- cyclohexen-1-one, caffeine and 5,6-dimethoxy-1(3 H) isobenzofuranone. This information allowed for the identification of nematocidal compounds in the degraded substrate of P. djamor, an activity that had not been reported previously.
Assuntos
Anti-Helmínticos , Haemonchus , Animais , Anti-Helmínticos/farmacologia , Antinematódeos/farmacologia , Antinematódeos/uso terapêutico , Larva , Extratos Vegetais/farmacologia , PleurotusRESUMO
During the previous decades, the indiscriminate use of anthelmintics for the control of gastrointestinal nematodes has generated anthelmintic resistance in different parts of the world. It is necessary to search for new sustainable control alternatives, such as the use of extracts from plants and edible mushrooms. Therefore, the aim of the present study was to evaluate the nematicidal activity of extracts and fractions of the edible mushroom Lentinula edodes against Haemonchus contortus eggs and infective larvae. The basidiomata of L. edodes ECS-401 were provided by the Tropical Fungi Laboratory of El Colegio de la Frontera Sur and were extracted with ethyl acetate, methanol, and water. Fractionation of the most active extract was carried out by open column chromatography. The bioassays were performed in 96-well microtiter plates using 100 eggs/larvae, a final volume of 100 µL, and different concentrations of extracts/fractions (n = 4). Bioassay readings were taken at 48 h for egg hatching inhibition (EHI) and at 24, 48 and 72 h for larval mortality (LM). The ethyl acetate extract exhibited the highest percentage of EHI (100%). For the LM bioassay, the aqueous extract was the most active (69%), but its fractions did not show larvicidal activity. The chemical profile of the aqueous extract was analyzed by high-performance liquid chromatography, which showed the presence of phenols, flavonoids, and terpenes. L. edodes extracts showed ovicidal and/or larvicidal activity.
Assuntos
Anti-Helmínticos , Haemonchus , Cogumelos Shiitake , Animais , Anti-Helmínticos/farmacologia , Flavonoides/farmacologia , Larva , Extratos Vegetais/farmacologiaRESUMO
Sheep haemonchosis is a disease that causes anemia, diarrhea, weight loss, low production, and even death of young animals. This represents a negative economic impact on the livestock sector. For decades, chemicals have been used to control this parasitic disease; however, cases of anthelmintic resistance have increased around the world. For this reason, the search for control alternatives is necessary. Several studies have shown that edible mushrooms of Pleurotus genus have different medicinal properties, including nematicidal activity. In this study, the chromatographic fractionation of the hydroalcoholic extract of fruiting bodies Pleurotus djamor was carried out to identify and isolate the metabolites responsible for nematicidal activity. From the fractionation of the extract, the fraction PdR2 and the subfraction PdB were obtained, which were evaluated against Haemonchus contortus under in vitro conditions. The nematicidal effect of the fraction PdB in gerbils infected artificially with H. contortus was evaluated. Finally, the analysis of the PdB fraction by gas chromatography-mass spectrometry and nuclear magnetic resonance was carried out. The results showed that the PdB had 100% of egg hatching inhibition from 5 mg/mL. Regarding larvicidal activity, it presented >97.2% from 24 h to 20 mg/mL. The in vivo evaluation of the PdB fraction showed a reduction of H. contortus larvae of 92.56%. The compounds present in this fraction were the mixture of allitol and an unidentified terpene in a ratio of 9:1. The PdB fraction is a potential alternative for the control of H. contortus, where allitol and a terpene could be responsible for nematicidal activity.
Assuntos
Anti-Helmínticos , Haemonchus , Pleurotus , Animais , Anti-Helmínticos/farmacologia , Larva , Extratos Vegetais/farmacologia , OvinosRESUMO
This study was aimed at evaluating the in vitro effect of the edible mushroom (EM) Pleurotus eryngii against the eggs and larvae (L3) of Haemonchus contortus. The evaluation included acetone (AE) and hydroalcoholic (HA) extracts of the following strains: ECS-1138, ECS-1156, ECS-1255, ECS-1258, ECS-1261, ECS-1282, and ECS-1292. The HA extract of the ECS-1255 strain showed the highest effect on mortality rates of L3 (18.83%) at 20 µg/mL. After subjecting this HA extract to a normal phase chromatography column, five fractions were obtained; fraction F5 (100% MeOH) was the most effective against eggs, with hatching inhibition percentages of 88.77 and 91.87% at 20 and 40 mg/mL, respectively. Gas chromatography-mass spectrometry (GC-MS) subjected this fraction to an acetylation reaction to determine the content of the secondary metabolites. The GC-MS analysis showed that the F5 fraction was composed of trehalose CAS: 6138-23-4, polyols (L-iditol CAS: 488-45-9, galactitol CAS: 608-66-2, D-mannitol CAS: 69-65-8, D-glucitol CAS: 50-70-4, and myoinositol CAS: 87-89-8), adipic acid CAS: 124-04-9, stearic acid CAS: 57-11-4, squalene CAS: 111-02-4, and ß-sitosterol CAS: 83-46-5.
Assuntos
Anti-Helmínticos/farmacologia , Ovos , Haemonchus/crescimento & desenvolvimento , Pleurotus/química , Animais , Anti-Helmínticos/química , Larva/crescimento & desenvolvimentoRESUMO
The goal of this study was to isolate, select and characterize bacteria with cellulolytic activity from two different coffee residue composting piles, one of which had an internal temperature of 57 -#9702;C and pH 5.5 and the other, a temperature of 61 -#9702;C, and pH 9.3. Culture media were manipulated with carboxymethylcellulose and crystalline cellulose as sole carbon sources. The enzyme activity was assessed by hydrolysis halo formation, reducing sugar production and zymograms. Three out of twenty isolated strains showed higher enzymatic activity and were identified as Bacillus subtilis according to their morphological, physiological, biochemical characteristics and based on the sequence analysis of 16S rDNA regions. The enzymatic extracts of the three selected strains showed exocellulase and endocellulase maximum activity of 0.254 and 0.519 U/ml, respectively; the activity of these enzymes was maintained even in acid pH (4.8) and basic (9.3) and at temperatures of up to 60°C. The enzymatic activities observed in this study are within the highest reported for cellulose produced by bacteria of the genus Bacillus. Endocellulase activity was shown in the zymograms from 24 h until 144 h of incubation. Furthermore, the pH effect on the endocellulase activity is reported for the first time by zymograms. The findings in this study entail the possibility to use these enzymes in the procurement of fermentable substrates for the production of energy from the large amount of residues generated by the coffee agroindustry.
El objetivo de este estudio fue aislar, seleccionary caracterizar bacterias con actividad celulolítica a partir de 2 diferentes pilas de compostaje de residuos de café, una con temperatura interna de 57°C y pH 5,5; la otra con temperatura interna de 61 °C y pH 9,3. Se utilizaron medios de cultivo con carboximetilcelulosa y celulosa cristalina como únicas fuentes de carbono. La actividad enzimàtica fue evaluada por formación de halos de hidrólisis, producción de azúcares reductores y zimogramas. De 20 cepas aisladas, 3 presentaron mayor actividad enzimàtica y fueron identificadas como Bacillus subtilis sobre la base de sus características morfológicas, fisiológicas y bioquímicas y del análisis de las secuencias de la región 16S del ADNr. Los extractos enzimáticos de las 3 cepas seleccionadas presentaron actividad de exocelulasa y de endocelulasa, con máximos de 0,254 y 0,519 U/ml, respectivamente; la actividad de estas enzimas se mantuvo incluso a pH ácido (4,8) o básico (9,3) y a temperaturas de hasta 60 °C. Las actividades enzimáticas halladas en este estudio se ubican dentro de las más altas reportadas para celulasas producidas por bacterias del género Bacillus. En los zimogramas se demostró actividad de endocelulasa desde las 24h hasta las 144h de incubación. Asimismo, se reporta por primera vez el efecto del pH sobre la actividad de endocelulasa observado por zimogramas. Los resultados de este estudio abren la posibilidad de hacer uso de estas enzimas en la obtención de sustratos fermentables para la producción de energía a partir de los residuos generados en grandes cantidades por la agroindustria del café.
Assuntos
Bacillus subtilis , Café , Celulases , Bacillus subtilis/isolamento & purificação , Bacillus subtilis/enzimologia , Compostagem , Celulose , Celulases/metabolismoRESUMO
In Older Adults (OAs), Electroencephalogram (EEG) slowing in frontal lobes and a diminished muscle atonia during Rapid Eye Movement sleep (REM) have each been effective tracers of Mild Cognitive Impairment (MCI), but this relationship remains to be explored by non-linear analysis. Likewise, data provided by EEG, EMG (Electromyogram) and EOG (Electrooculogram)-the three required sleep indicators-during the transition from REM to Non-REM (NREM) sleep have not been related jointly to MCI. Therefore, the main aim of the study was to explore, with results for Detrended Fluctuation Analysis (DFA) and multichannel DFA (mDFA), the Color of Noise (CN) at the NREM to REM transition in OAs with MCI vs. subjects with good performances. The comparisons for the transition from NREM to REM were made for each group at each cerebral area, taking bilateral derivations to evaluate interhemispheric coupling and anteroposterior and posterior networks. In addition, stationarity analysis was carried out to explore if the three markers distinguished between the groups. Neuropsi and the Mini-Mental State Examination (MMSE) were administered, as well as other geriatric tests. One night polysomnography was applied to 6 OAs with MCI (68.1 ± 3) and to 7 subjects without it (CTRL) (64.5 ± 9), and pre-REM and REM epochs were analyzed for each subject. Lower scores for attention, memory and executive funcions and a greater index of arousals during sleep were found for the MCI group. Results confirmed that EOGs constituted significant markers of MCI, increasing the CN for the MCI group in REM sleep. The CN of the EEG from the pre-REM to REM was higher for the MCI group vs. the opposite for the CTRL group at frontotemporal areas. Frontopolar interhemispheric scaling values also followed this trend as well as right anteroposterior networks. EMG Hurst values for both groups were lower than those for EEG and EOG. Stationarity analyses showed differences between stages in frontal areas and right and left EOGs for both groups. These results may demonstrate the breakdown of fractality of areas especially involved in executive functioning and the way weak stationarity analyses may help to distinguish between sleep stages in OAs.
RESUMO
The goal of this study was to isolate, select and characterize bacteria with cellulolytic activity from two different coffee residue composting piles, one of which had an internal temperature of 57°C and pH 5.5 and the other, a temperature of 61°C, and pH 9.3. Culture media were manipulated with carboxymethylcellulose and crystalline cellulose as sole carbon sources. The enzyme activity was assessed by hydrolysis halo formation, reducing sugar production and zymograms. Three out of twenty isolated strains showed higher enzymatic activity and were identified as Bacillus subtilis according to their morphological, physiological, biochemical characteristics and based on the sequence analysis of 16S rDNA regions. The enzymatic extracts of the three selected strains showed exocellulase and endocellulase maximum activity of 0.254 and 0.519 U/ml, respectively; the activity of these enzymes was maintained even in acid pH (4.8) and basic (9.3) and at temperatures of up to 60°C. The enzymatic activities observed in this study are within the highest reported for cellulose produced by bacteria of the genus Bacillus. Endocellulase activity was shown in the zymograms from 24h until 144h of incubation. Furthermore, the pH effect on the endocellulase activity is reported for the first time by zymograms. The findings in this study entail the possibility to use these enzymes in the procurement of fermentable substrates for the production of energy from the large amount of residues generated by the coffee agroindustry.
Assuntos
Bacillus subtilis , Celulases , Café , Bacillus subtilis/enzimologia , Bacillus subtilis/isolamento & purificação , Celulases/metabolismo , Celulose , CompostagemRESUMO
Fifty-four macromycetes, isolated from southeastern Mexico, were used in order to evaluate their capacity for degradation and tolerance to the herbicide paraquat. Ten of these strains were capable of growing in a solid culture medium in the presence of 200 ppm paraquat. Subsequently, assays to evaluate the degradation of the xenobiotic in a liquid medium were carried out. Of the ten strains evaluated, three presented the highest levels of degradation of the compound, which were Trametes pavonia (54.2%), Trametes versicolor (54.1%) and Hypholoma dispersum. They presented the highest overall degradation percentage (70.7%) after 12 days culture. The presence of ligninolytic enzymes in these strains was evaluated. H. dispersum only presented aryl alcohol oxidase activity; however, with the data obtained, it was not possible to conclude whether this specific enzyme is responsible for paraquat degradation. The level of degradation obtained is above the one reported for Pseudomonas putida, one of the few reports on paraquat degradation. This is the first report on the contaminant degradation capacity of H. dispersum.