9 research outputs found

    Fungal inoculation induces agarwood in young Aquilaria malaccensis trees in the nursery

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    Fungi are often used to induce agarwood in Aquilaria trees. This study was conducted to evaluate the effect of several fungi on agarwood formation over time in young Aquilaria malaccensis (Lam.) trees. Typical changes in the length and light intensity of the resulting discoloration were observed after three and six month periods following inoculation. Wood samples were observed microscopically and classified into several light intensity groups. The discoloration length was measured longitudinally. The duration after inoculation affected the mean of discoloration length: the 6-month old sample (1.70 cm) had a wider discoloration zone when compared to the 3-month old sample (1.17 cm). When measuring the discoloration intensity, a positive relationship with time was perceived. Digital images, captured using a camera-equipped microscope, revealed that wood samples collected after six months appeared to be 1.8-times darker than after three months. We concluded that time, not the species of any of the tested fungi, had significant effect on discoloration length and intensity. Gas chromatography/mass spectrometry (GCMS) analysis of the 6-month old sample yielded some important agarwood compounds such as benzylacetone, anisylacetone, guaiene and palustrol. This demonstrates that the tested fungi have the ability to induce agarwood formation in nursery A. malaccensis trees

    The Changing Landscape for Stroke\ua0Prevention in AF: Findings From the GLORIA-AF Registry Phase 2

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    Background GLORIA-AF (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients with Atrial Fibrillation) is a prospective, global registry program describing antithrombotic treatment patterns in patients with newly diagnosed nonvalvular atrial fibrillation at risk of stroke. Phase 2 began when dabigatran, the first non\u2013vitamin K antagonist oral anticoagulant (NOAC), became available. Objectives This study sought to describe phase 2 baseline data and compare these with the pre-NOAC era collected during phase 1. Methods During phase 2, 15,641 consenting patients were enrolled (November 2011 to December 2014); 15,092 were eligible. This pre-specified cross-sectional analysis describes eligible patients\u2019 baseline characteristics. Atrial fibrillation disease characteristics, medical outcomes, and concomitant diseases and medications were collected. Data were analyzed using descriptive statistics. Results Of the total patients, 45.5% were female; median age was 71 (interquartile range: 64, 78) years. Patients were from Europe (47.1%), North America (22.5%), Asia (20.3%), Latin America (6.0%), and the Middle East/Africa (4.0%). Most had high stroke risk (CHA2DS2-VASc [Congestive heart failure, Hypertension, Age  6575 years, Diabetes mellitus, previous Stroke, Vascular disease, Age 65 to 74 years, Sex category] score  652; 86.1%); 13.9% had moderate risk (CHA2DS2-VASc = 1). Overall, 79.9% received oral anticoagulants, of whom 47.6% received NOAC and 32.3% vitamin K antagonists (VKA); 12.1% received antiplatelet agents; 7.8% received no antithrombotic treatment. For comparison, the proportion of phase 1 patients (of N = 1,063 all eligible) prescribed VKA was 32.8%, acetylsalicylic acid 41.7%, and no therapy 20.2%. In Europe in phase 2, treatment with NOAC was more common than VKA (52.3% and 37.8%, respectively); 6.0% of patients received antiplatelet treatment; and 3.8% received no antithrombotic treatment. In North America, 52.1%, 26.2%, and 14.0% of patients received NOAC, VKA, and antiplatelet drugs, respectively; 7.5% received no antithrombotic treatment. NOAC use was less common in Asia (27.7%), where 27.5% of patients received VKA, 25.0% antiplatelet drugs, and 19.8% no antithrombotic treatment. Conclusions The baseline data from GLORIA-AF phase 2 demonstrate that in newly diagnosed nonvalvular atrial fibrillation patients, NOAC have been highly adopted into practice, becoming more frequently prescribed than VKA in Europe and North America. Worldwide, however, a large proportion of patients remain undertreated, particularly in Asia and North America. (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients With Atrial Fibrillation [GLORIA-AF]; NCT01468701

    Effects of mechanical wounding and infection patterns of fusarium solani on gaharu formation in Aquilaria malaccensis Lam

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    Gaharu is the oleoresin found in karas (Aquilaria malaccensis). Recently, many studies have been conducted in order to produce the most suitable and effective inocula in the market. Indeed, there is a need to produce more gaharu in karas tree especially in plantation field. Various fungi have been associated with gaharu formation, however their roles and patterns of colonization are not fully understood. In general, gaharu formation is associated with natural wounding or artificial inoculation by different fungi or chemicals. This study focused on the patterns of Fusarium solani colonization at different wounding time-point via real - time quantitative PCR (qPCR) technique and the effects of artificial inoculation on young karas trees as compared to naturally infected wounded karas in the wild. In addition, artificial inoculation technique with different fungal isolates were also aimed to determine their effects on producing gaharu. This was done by comparing their degree of discoloration formed over a period of 1, 3, 6 months respectively and the chemical constituents. Gaharu compounds produced from both induction methods were identified and compared to commercial samples via GCMS. When tracking the amount of F. solani inocula in two wounded tree over time in the wild, the fungus was found to colonize wounded karas most frequently at the early stage of wounding, often most abundantly during the first 2 weeks. The highest amount of target DNA copies was quantified at 2 weeks in both investigated trees. This indicates that the infection of fungi might take place at the early stage of the wounding process. The decreasing in gene copy number detected in the latter stages, from 6 to 12 months showed that there was no evidence to state that fungi colonized more abundantly in darker wood. In fact, more resins were produced in order to form gaharu. The high level of terpenes were maintained to control the pathogen attacked. The presence of F. solani consistently at every wounding time - point, except for two points in one of the trees, revealed the association of this microbe to gaharu formation. However, its direct role in gaharu formation has to be further investigated, perhaps in a more controlled environment. The study on fungal inoculation was carried out on young 4-year-old trees in a shade house conditions. Trees were harvested at 1, 3 and 6 months after inoculation. The discoloration on the wound and its intensity were measured. ANOVA analysis concluded that the five species of fungal inocula tested were not giving significant differences in terms of discoloration length and intensity. However, the length of discoloration and its intensity increased progressively from pale yellow to dark brown over the time period tested. This implies that time plays an important role for producing gaharu-impregnated wood. When analyzing gaharu compounds using GC-MS, a total of 39 compounds mainly from the group monoterpenes,sesquiterpenes, oxygenated sesquiterpenes, chromone derivatives and fatty acids were identified. Samples analyzed were from commercial gaharu, artificial inoculation and mechanically wounded wood. From GC-MS profiles, all five commercial gaharu, SA, A, B, C and D grades had similar important compounds. Among them were (-)-Aristolene, Palustrol (derivatives of guaiene), γ. Eudesmol, (-)-Aristolene, α-Elemol, Anisylacetone, Benzaldehyde, Benzylacetone, 6-(Benzyloxy)-4, 4-dimethyl-2- chromaone and Agarospirol. When compared to artificial inoculated wood,there were some important fragrant compounds found such as Guaiene, Benzenepropanoic acid,2,5-dimethoxy, Palustrol, Benzylacetone, Benzaldehyde, 6-(Benzyloxy)-4,4-dimethyl-2-chromaone. A new compound of 6-(Benzyloxy)-4,4-dimethyl-2- chromaone was detected in the wood sample and it was also known as 4,4-dimethyl - 6 - phenylmethoxy-3H-chromen-2-one. In overall, the existence of these compounds proved that gaharu was formed in the fungal inoculated wood. This study demonstrated that fungal inoculation could produce gaharu in young trees in just 6 months time. Fungal invasion could be a method to prolong agression since it could indirectly increase resin production. However, this should be tested on a larger scale under field trial conditions

    Gas Chromatography-Mass Spectrometry Analysis of Agarwood Extracts from Mature and Juvenile Aquilaria malaccensis

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    International audienceChemical composition of crude extracts from infected woods of Aquilaria malaccensis were compared to that of healthy wood and commercial agarwood. Infected woods were collected six months after drilling of wild mature trees or after fungal inoculation into the stem of 4-year-old trees. Agarwood substances were extracted in methanol and were subjected to GC-MS analyses. The major compounds were chromone derivative, aromatic compounds, sesquiterpenes, monoterpenes, sterols and fatty acid methyl ester. Aromatic compounds constituted of aldehyde, phenol, ether and ketone groups. In the agarwood extract of the juvenile fungal-elicited tree but not in the healthy wood, some major compounds found were 2-(2-phenylethyl) chromone derivative, 4-phenyl-2-butanone, (1S, 4S, 7R)-1,4-dimethyl-7-(prop-1-en-2-yl)-1,2,3,4,5,6,7,8-octahydroazulene [guaiene], 1,1,4,7-tetramethyl-2,3,4,5,6,7,7a, 7b-octahydro-1aH-cyclopropa[h]azulen-4a-ol [palustrol], and 4-(4-methoxyphenyl) butan-2-one [anisylacetone]. These were also found from agarwood of different grades and agarwood collected from the wild mature tree, in addition to agarospirol, alloaromadendre oxide (2), alpha-elemol, gamma-eudesmol, and guaiol. This work demonstrated that in young A. malaccensis trees, fungi may be associated to the formation of important agarwood compounds and can be detected as early as six months after inoculation

    Succession patterns of fungi associated to wound-induced agarwood in wild Aquilaria malaccensis revealed from quantitative PCR assay

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    Aquilaria malaccensis produces agarwood in response to wounding and fungal attack. However, information is limited regarding Aquilaria‘s interaction with its diverse fungal community. In this study, time-related changes of three natural fungal colonizers in two wounded wild A. malaccensis were tracked, beginning a few hours after wounding up to 12 months. Using species-specific primers derived from their nrITS sequences in quantitative real-time PCR (qPCR), we quantified the amount of Cunninghamella bainieri, Fusarium solani and Lasiodiplodia theobromae. Because time is a major factor affecting agarwood quantity and quality, 14 wood samples were collected at different time points, i.e., 0–18 h, 2–13 days, 2–18 weeks, and 6–12 months after wounding. qPCR data revealed that the abundance of the three species decreased over time. The fungi were detected in high numbers during the first few hours and days after wounding (40- to 25,000-fold higher levels compared with initial counts) and in low numbers (<1- to 3,200-fold higher than initially) many months later. Consistent with its role in defense response, the accumulation of secondary metabolites at the wounding site could have caused the decline in fungal abundance. Succession patterns of the two trees were not identical, indicating that fungal populations may have been affected by tree environment and wound microclimate. Our results are important for understanding the diversity of microbial community in wild Aquilaria species and their association to wound-induced agarwood formation. Fungi could be secondary triggers to agarwood production in situations where trees are wounded in attempt to induce agarwood

    Identification of Fungal Communities in "Agarwood' from wild Aquilaria malaccensis using rDNA Spacer Sequences

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    Aquilaria malaccensis is a medium-sized tropical tree from the family Thymelaeaceae, and is the main 'agarwood' producer in Peninsular Malaysia. 'Agarwood' is a valuable fragrant resin, produced in the tree trunks of A. ma/accensis, often associated to pathological infections by fungi. In our attempt to understand the roles of fungi in 'agarwood' production, we isolated fungi from 'agarwood', collected from a wild A. ma/accensis population in Peninsular Malaysia. The fungal isolates were from the genera Fusarium, Trichoderma, Curvularia and Cunninghamella (order Mucorales) based on culture characteristics on agar medium. To confirm the genera, we analyzed genomic DNA from pure cultures and wood samples from which they were first isolated., ' Using universal primers in PCR, we amplified the intemal transcribed spacer (ITS) within the ribosomal DNA (rONA) regions. Spacer sequences provided supporting evidence to the genera designation via the culture-based method. The method promised a rapid detection and identification of fungal communities directly from environmental and wood samples

    The Changing Landscape for Stroke\ua0Prevention in AF

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    The Changing Landscape for Stroke Prevention in AF

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