Tropical forest plants are known to be pollinated by a diverse array of animals. Here we report on the pollination of a woody climber species, Uvaria elmeri (Annonaceae), by cockroaches in a lowland mixed-dipterocarp forest in Sarawak, Malaysia. To the best of our knowledge, this is the first report of pollination by cockroaches. The cauliflorous flowers in the understory are protogynous and bloomed for 50 h. An odor similar to decayed wood or a mushroom was secreted by flowers and was stronger during the male stage. Pollinators were cockroaches (Blattellidae) and drosophilid flies (Drosophilidae). Cockroaches, the main pollinators, visited flowers during both female and male stages at night, feeding on stigmatic exudate and pollen. Drosophilids, the secondary pollinators. mainly visited female-stage flowers during daytime, fed on stigmatic exudate. and laid eggs on stigmas. Neither autogamy nor self-compatibility was observed. Fruit production appeared to be pollen-limited. The fruit set, which was 2% of flowers in natural condition, was significantly lower than the 30% fruit set obtained by artificial cross-pollination. We discuss the traits of cockroaches as pollinators and the breeding system of U. elmeri.
DNA barcoding involves the use of one or more short, standardized DNA fragments for the rapid identification of species. A 648-bp segment near the 5' terminus of the mitochondrial cytochrome c oxidase subunit I (COI) gene has been adopted as the universal DNA barcode for members of the animal kingdom, but its utility in mushrooms is complicated by the frequent occurrence of large introns. As a consequence, ITS has been adopted as the standard DNA barcode marker for mushrooms despite several shortcomings. This study employed newly designed primers coupled with cDNA analysis to examine COI sequence diversity in six species of Pleurotus and compared these results with those for ITS. The ability of the COI gene to discriminate six species of Pleurotus, the commonly cultivated oyster mushroom, was examined by analysis of cDNA. The amplification success, sequence variation within and among species, and the ability to design effective primers was tested. We compared ITS sequences to their COI cDNA counterparts for all isolates. ITS discriminated between all six species, but some sequence results were uninterpretable, because of length variation among ITS copies. By comparison, a complete COI sequences were recovered from all but three individuals of Pleurotus giganteus where only the 5' region was obtained. The COI sequences permitted the resolution of all species when partial data was excluded for P. giganteus. Our results suggest that COI can be a useful barcode marker for mushrooms when cDNA analysis is adopted, permitting identifications in cases where ITS cannot be recovered or where it offers higher resolution when fresh tissue is. The suitability of this approach remains to be confirmed for other mushrooms.
Intensive land expansion of commercial oil palm agricultural lands results in reducing the size of peat swamp forests, particularly in Southeast Asia. The effect of this land conversion on macrofungal biodiversity is, however, understudied. We quantified macrofungal biodiversity by identifying mushroom sporocarps throughout four different habitats; logged peat swamp forest, large-scale oil palm plantation, monoculture, and polyculture smallholdings. We recorded a total of 757 clusters of macrofungi belonging to 127 morphospecies and found that substrates for growing macrofungi were abundant in peat swamp forest; hence, morphospecies richness and macrofungal clusters were significantly greater in logged peat swamp forest than converted oil palm agriculture lands. Environmental factors that influence macrofungi in logged peat swamp forests such as air temperature, humidity, wind speed, soil pH, and soil moisture were different from those in oil palm plantations and smallholdings. We conclude that peat swamp forests are irreplaceable with respect to macrofungal biodiversity. They host much greater macrofungal biodiversity than any of the oil palm agricultural lands. It is imperative that further expansion of oil palm plantation into remaining peat swamp forests should be prohibited in palm oil producing countries. These results imply that macrofungal distribution reflects changes in microclimate between habitats and reduced macrofungal biodiversity may adversely affect decomposition in human-modified landscapes.
Mushrooms have been highly regarded as possessing enormous nutritive and medicinal values. In the present study, we evaluated the anti-oxidative and anti-atherosclerotic potential of shiitake mushroom (Lentinula edodes) using its solvent-solvent partitioned fractions that consisted of methanol:dichloromethane (M:DCM), hexane (HEX), dichloromethane (DCM), ethyl acetate (EA) and aqueous residue (AQ). The hexane fraction (1 mg/mL) mostly scavenged (67.38%, IC50 0.55 mg/mL) the 2,2-diphenyl-1-picryl hydrazyl (DPPH) free radical, contained the highest reducing capacity (60.16 mg gallic acid equivalents/g fraction), and most potently inhibited lipid peroxidation (67.07%), low density lipo-protein oxidation and the activity of 3-hydroxy 3-methyl glutaryl co-enzyme A reductase (HMGR). GC-MS analyses of the hexane fraction identified α-tocopherol (vitamin E), oleic acid, linoleic acid, ergosterol and butyric acid as the bio-functional components present in L. edodes. Our findings suggest that L. edodes possesses anti-atherosclerotic bio-functionality that can be applied as functional food-based therapeutics against cardiovascular diseases.
The aim of this study was to determine the effect of different ratios of low protein flour to oyster mushroom (Pleurotus sajor-caju) powder on the physicochemical properties and sensory acceptability of edible tablespoon. Fresh grey oyster mushroom was dried in a convection oven at temperature of 55.0˚C ± 2.0˚C for 20 h prior to the grinding process. The low protein flour (LPF) was then incorporated with oyster mushroom powder (OMP) at different ratios of 100:0, 96:4, 92:8, 88:12 and 84:16, before being with vegetable oil, sugar, egg white and water in formulating the edible tablespoon. The proximate analyses were carried out in triplicate for calorie content, colour profile, hardness value and morphological structure of edible tablespoon. This study revealed that with decreasing LPF and increasing OMP in the formulation, the ash content (1.24% to 1.92%), crude fat content (8.98% to 10.40%) and fiber content (0.13% to 1.24%) were observed to have increased as well as the hardness value (2042.03g to 2844.57g) and pore’s size of the morphological structure of edible tablespoon. However, the carbohydrate content (78.64% to 75.56%) significantly decreased (p>0.05) together with L* value (from 68.47 to 61.71) when the decrease was in the the percentage of LPF and an increase the percentage of OMP. The calorie content, moisture content and protein content of edible tablespoon were not significantly (p>0.05) affected by different ratios of LPF to OMP. The edible tablespoon formulated with up to 8% of OMP was accepted by the sensory panelists but further increase in OMP addition significantly decreased the degree of likeness in terms of colour, odour, taste and overall acceptability of edible tablespoon. This study suggested that oyster mushroom edible tablespoon could be potential alternative disposable cutlery which will help to reduce the use of huge amount of non-biodegradable materials for environmental conservation.
Introduction: Hygrocybe conica (HC), a wild mushroom commonly consumed by the indigenous people (Orang Asli) in Peninsular Malaysia, was assessed for its antioxidant content. Methods: The HC mushroom was extracted using distilled water and the crude extract partitioned using different solvents and open column chromatography to evaluate its potential antioxidant properties. The mushroom extract was partitioned using liquid-liquid extraction into the hexane (Fl), chloroform (F2), butanol (F3) and formic acid (F4) fractions. Based on solvent polarity, the water extract of the mushroom was fractionated into non-polar (FI), semi-polar (Fii), and polar fractions (Fiii) using open column chromato graphy. Antioxidant capacities were determined using DPPH, ABTS, and ferric reducing antioxidant power (FRAP) assays while Folin-Ciocalteu reagent assay was used to determine total phenolic content (TPC). Results: The HC extract had the highest TPC and DPPH scavenging capacity compared to its extract fractions. TE values (ABTS assay) of F2 and F4 were not significantly higher than the HC extract. Among the extract fractions of different polarities, Fiii had the highest antioxidant capacities (DPPH and FRAP) compared to FI and Fii while FRAP values of these fractions were not significantly lower than the FRAP value of HC extract. The HC extract had significantly lower antioxidant capacity than antioxidant standards (ascorbic acid and BHA). Tannie acid as the main bioactive component in HC mushroom was detected using HPLC method. The presence of phenolics in HC extract was also confirmed using TLC. Conclusion: Due to the presence of potent phenolic components, the mycelia of HC could be consumed for potential antioxidative benefits.
Over the years, natural products have shown success as antidiabetics in in vitro, in vivo studies and clinical trials. Because natural product-derived drugs are more affordable and effective with fewer side-effects compared to conventional therapies, pharmaceutical research is increasingly leaning towards the discovery of new antidiabetic drugs from natural products targeting pathways or components associated with type 2 diabetes mellitus (T2DM) pathophysiology. However, the drug discovery process is very lengthy and costly with significant challenges. Therefore, various techniques are currently being developed for the preclinical research phase of drug discovery with the aim of drug development with less time and efforts from natural products. In this review, we have provided an update on natural products including fruits, vegetables, spices, nuts, beverages and mushrooms with potential antidiabetic activities from in vivo, in vitro and clinical studies. Synergistic interactions between natural products and antidiabetic drugs, and potential antidiabetic active compounds from natural products are also documented to pave the way for combination treatment and new drug discovery, respectively. Additionally, a brief idea of the drug discovery process along with the challenges that arise during drug development from natural products and the methods to conquer those challenges are discussed to create a more convenient future drug discovery process.
Tiger’s Milk mushroom has been used for medicinal purposes by local aborigines to treat asthma, breast cancer, cough, fever and food poisoning. Molecular phylogenetic analysis utilizing RNA polymerase II, second largest subunit (RPB2) gene, identified the wild Tiger’s Milk mushrooms collected from the state of Pahang in Malaysia for this study as Lignosus rhinocerus in the order Polyporales. The tuber, stipe and pileus of this mushroom were analyzed for their basic nutritional composition (fat, protein, and carbohydrate) and toxic metal content profile (Cadmium, Lead and Mercury). The moisture content of these mushroom parts varied from 32.22% (pileus) – 46.31% (stipe). The dry matter of the mushrooms contained 2.76% (stipe) – 6.60% (pileus) proteins, 0.21% (pileus) – 0.30% (tuber) fat, 1.76% (stipe) – 4.38% (tuber) ash and 38.47% (stipe) – 56.30% (pileus) carbohydrates. The toxic metal content of the mushroom samples ranged from 0.03–0.12 mg/kg for Cd, 0.80–1.94 mg/kg for Pb and 0.05–0.10 mg/kg for Hg. The present study demonstrated that L. rhinocerus is a potential source of food due to its high carbohydrate content. In addition, the trace levels of toxic metals in this mushroom are within the safe level for consumption.
Lignosus also known as "Tiger Milk Mushroom," is classified in the family Polyporaceae and mainly consumed for its medicinal properties in Southeast Asia and China. The sclerotium is known as the part with medicinal value and often used by the natives to treat a variety of ailments. Lignosus tigris Chon S. Tan, one of the species of the Malaysia Tiger Milk mushroom, has recently been successfully cultivated in laboratory. Earlier studies have demonstrated the L. tigris cultivar E sclerotia exhibited beneficial biomedicinal properties. This study evaluated the potential toxicity of L. tigris E sclerotia in a 28-day sub-acute oral administration in Sprague Dawley (SD) rats. L. tigris E sclerotial powder was administered orally at three different doses of 250, 500, and 1000 mg/kg to the SD rats once daily, consecutively for 28-days. Body weight of the rats was recorded and general behavior, adverse effects, and mortality were observed daily throughout the experimental period. At the end of the experiment, blood hematology and biochemistry, relative organ weights, and histopathological analysis were performed. Results showed that there were no mortality nor signs of toxicity throughout the 28-day sub-acute toxicity study. Oral administration of the L. tigris E sclerotial powder at daily dose up to 1000 mg/kg had no significant effects in body weight, relative organ weight, blood hematological and biochemistry, gross pathology, and histopathology of the organs. L. tigris E sclerotial powder did not cause any treatment-related adverse effect in the rats at different treatment dosages up to 1000 mg/kg. As the lethal dose for the rats is above 1000 mg/kg, the no-observed-adverse-effect level (NOAEL) dose is more than 1000 mg/kg.
Termitomyces are delicious edible mushrooms found in Africa and South-East Asia including Malaysia. These mushrooms were found to grow symbiotically with termites around termite nests. Numerous efforts have been made worldwide to develop a cultivation method for these mushrooms. Unfortunately, none of those attempts were successful. The main obstacles encountered were the difficulty to identify and isolate pure termitomyces culture. The problem became prevalent as the culture gets contaminated by other fungi. Termitomyces can easily be identified by its mushroom fruiting body eventually but certainly not at the mycelium and hyphea stages. In this study a simple PCR-based genetic marker detection method for confirmation of termitomyces at any culture stage was developed. Using this method, four distinctive PCR assays
were developed using specific PCR primers designed based on the DNA sequence of the termitomyces mushroom. The PCR results showed that the PCR assays using intact termitomyces
DNA as template was not suitable for this purpose. However, PCR using BamHI and EcoRI predigested termitomyces DNA as template showed identical polymorphism pattern for both
termitomyces mushroom DNA and termitomyces culture DNA. Thus, the method reported here can be used for the identific.
Discovery of novel dietary fibre (DF)-rich food ingredient is of great interest to meet the rising consumer demand for healthy food. Intake of DF-enriched food has been positively associated with a decreased risk of chronic diseases. Pleurotus sajor-caju (PSC), one of the prominent edible mushrooms widely cultivated in Malaysia, is useful for its culinary and medicinal purpose. This study aimed to determine the nutritional, physical, colour and sensorial properties of cinnamon biscuit formulated with PSC powder as partial replacement (0, 4, 8 and 12%) for wheat flour. Results indicated that elevated incorporation levels of PSC powder significantly (P
A 65-year-old lady complained of occasional flashes of light over her left eye for 2 months. She was referred for ocular assessment after she underwent refractive assessment at the optometrist. On examination, best corrected visual acuity in both eyes was 6/9. Both eyes anterior segments were normal, with normal intraocular pressure. The pupils were equal with no relative afferent pupillary defect. Left eye fundus examination was normal. On the contrary, the "normal" right eye fundus examination revealed a huge, well-circumscribed hypo-pigmented elevated choroidal mass. B scan ultrasonography of the right eye showed a mushroom-shaped intraocular mass. Magnetic resonance imaging showed an intraocular lesion. Comprehensive systemic examination and investigations to rule out distant primary malignancy were unremarkable. A provisional diagnosis of right eye primary choroidal melanoma was made. Despite good vision in the affected eye, she underwent right eye enucleation in view of the large intraocular mass. The diagnosis was confirmed by histopathological examination.
In the current research work, different N-(substituted-phenyl)-4-{(4-[(E)-3-phenyl-2-propenyl]-1-piperazinyl}butanamides have been synthesized according to the protocol described in scheme 1. The synthesis was initiated by reacting various substituted anilines (1a-e) with 4-chlorobutanoyl chloride (2) in aqueous basic medium to give various electrophiles, 4-chloro-N-(substituted-phenyl)butanamides (3a-e). These electrophiles were then coupled with 1-[(E)-3-phenyl-2-propenyl]piperazine (4) in polar aprotic medium to attain the targeted N-(substituted-phenyl)-4-{(4-[(E)-3-phenyl-2-propenyl]-1-piperazinyl}butanamides (5a-e). The structures of all derivatives were identified and characterized by proton-nuclear magnetic resonance (1H NMR), carbon-nuclear magnetic resonance (13C NMR) and Infra-Red (IR) spectral data along with CHN analysis. The in vitro inhibitory potential of these butanamides was evaluated against Mushroom tyrosinase, whereby all compounds were found to be biologically active. Among them, 5b exhibited highest inhibitory potential with IC50 value of 0.013 ± 0.001 µM. The same compound 5b was also assayed through in vivo approach, and it was explored that it significantly reduced the pigments in zebrafish. The in silico studies were also in agreement with aforesaid results. Moreover, these molecules were profiled for their cytotoxicity through hemolytic activity, and it was found that except 5e, all other compounds showed minimal toxicity. The compound 5a also exhibited comparable results. Hence, some of these compounds might be worthy candidates for the formulation and development of depigmentation drugs with minimum side effects.
The effect of replacement of ricebran by mushroom stalk (Pleurotus florida), an agriculture waste on growth performance of African catfish (Clarias gariepinus) fingerlings was investigated for 42 days. Three isonitrogenous (32% crude protein) experimental diets containing 100% (Diet 1), 50% (Diet 2) and 0% (Diet 3) mushroom meal were formulated. Each diet was randomly allocated to triplicate groups of fingerlings in experimental tanks, with each tank stocking 12 fishes. The fishes were fed twice daily to apparent satiation for 6 weeks. Good survival rates were obtained from fish fed with Diet 1 (94%) and Diet 3 (94%). There was no significant differences in body weight gain (BWG), specific growth rate (SGR) and feed conversion ratio (FCR) among all diets (p>0.05). The fish fed with Diet 1 showed the best BWG, SGR, FCR and PER compared with Diet 2 and Diet 3. An economic evaluation indicated that Diet 1 gave the lowest production cost at RM2.03/kg followed by Diet 2 and Diet 3 at RM2.18/kg and RM2.38/kg, respectively. This study showed that the best diet was Diet 1 with 100% replacement of rice bran with mushroom stalks costing RM2.03/kg, the cheapest among the three diets.
Mushrooms are basically fungi, which have fleshy and spore-bearing fruiting body. This family of fungi literally has thousands of varieties of mushroom throughout the world. Oyster mushrooms are uniquely distinctive and they do look like oysters. Drying these mushroom confer a stabilizing property to it and then can be stored for a longer period. The nutritional values of the dried oyster mushroom with different drying techniques were thus determined. There were three different drying techniques used. These include low heat air blow (LHAB, AnjaadTM), sun drying (SD) and gas laboratory oven (LO) drying. All three samples were analyzed for beta-glucan content, water activity, colour, proximate analysis and dietary fibre concentration. The result showrd that LHAB method confers the lowest water activity compared with the other two drying methods. It also has the lowest colour measurement for brightness. Mushroom samples dried by LHAB techniques contain the highest concentration of both fat and carbohydrate compared with the other two methods. Besides, SD method confers the highest beta-glucan content. On the other hand, dietary fibres observed in LO dried samples contain the highest fibre content among the three drying treatments. In conclusion, LHAB method is recommended in reducing water activity and increasing proximate contents while both SD and LO are good in preserving beta-glucan and dietary fibre contents, respectively.
Twenty-eight days subacute toxicity studies performed in rats using sclerotial powder of Lignosus cameronensis cultivar was conducted to assess its safety for consumption prior to other scientific investigations on its medicinal benefits, nutraceutical or pharmaceutical application of the mushroom. The study was conducted at 250, 500, and 1000 mg/kg sclerotial powder of L. cameronensis cultivar (n = 5 for each respective dose, on both male and female groups) while control groups received only distilled water. At the end of the study (29th day), the animals were sacrificed followed by blood and organs collection for analysis. Subacute toxicity studies done shows that sclerotial powder of L. cameronensis cultivar at 250, 500, and 1000 mg/kg did not induce treatment related changes on behavioral patterns, gross physical appearance, growth pattern, body weight gain, values of hematological and clinical biochemical panels as well as histopathological findings on kidney, spleen, heart, lung and liver of the experimental rats. The no-observed-adverse-effect level dose for sclerotial powder of L. cameronensis cultivar in 28-days sub-acute toxicity study is determined to be 1000 mg/kg.
Edible and medicinal mushrooms are regularly used in natural medicines and home remedies since antiquity for ailments like fever, inflammation, and respiratory disorders. Lignosus rhinocerotis (Cooke) Ryvarden is a polypore found in Malaysia and other regions in South East Asia. It can be located on a spot where a tigress drips milk while feeding, hence the name "tiger's milk mushroom." The sclerotium of L. rhinocerotis is highly sought after by the native communities in Malaysia to stave off hunger, relieve cough and asthma, and provide stamina. The genomic features of L. rhinocerotis have been described. The pharmacological and toxicity effects, if any, of L. rhinocerotis sclerotium have been scientifically verified in recent years. In this review, the validated investigations including the cognitive function, neuroprotection, immune modulation, anti-asthmatic, anti-coagulation, anti-inflammatory, anti-microbial/ anti-viral, anti-obesity, anti-cancer/ anti-tumor, and antioxidant properties are highlighted. These findings suggest that L. rhinocerotis can be considered as an alternative and natural medicine in the management of non-communicable diseases. However, there is a paucity of validation studies including human clinical trials of the mycochemicals of L. rhinocerotis.
A national treasure mushroom, Lignosus rhinocerus, has been used to treat variety of ailments by local and indigenous communities in Malaysia. The aim of this study was to investigate the potential of the most valuable part of L. rhinocerus, the sclerotium, on neurite outgrowth activity by using PC-12Adh cell line. Differentiated cells with one thin extension at least double the length of the cell diameter were scored positive. Our results showed that aqueous sclerotium L. rhinocerus extract induced neurite outgrowths of 24.4% and 42.1% at 20 μg/mL (w/v) of aqueous extract alone and a combination of 20 μg/mL (w/v) aqueous extract and 30 ng/mL (w/v) of NGF, respectively. Combination of NGF and sclerotium extract had additive effects and enhanced neurite outgrowth. Neuronal differentiation was demonstrated by indirect immunofluorescence of neurofilament protein. Aqueous sclerotium extract contained neuroactive compounds that stimulated neurite outgrowth in vitro. To our knowledge this is the first report on neurite-stimulating activities of L. rhinocerus.
Lignosus rhinocerus, the tiger milk mushroom, is one of the most important medicinal mushrooms used by the indigenous people of Southeast Asia and China. It has been used to treat breast cancer. A cold water extract (LR-CW) prepared from the sclerotia of L. rhinocerus cultivar was found to exhibit antiproliferative activity against human breast carcinoma (MCF-7) and human lung carcinoma (A549), with IC(50) of 96.7 μg/mL and 466.7 μg/mL, respectively. In comparison, LR-CW did not show significant cytotoxicity against the two corresponding human normal cells, 184B5 (human breast cell) and NL 20 (human lung cell). DNA fragmentation studies suggested that the cytotoxic action of LR-CW against cancer cells is mediated by apoptosis. Sephadex G-50 gel filtration fractionation of LR-CW yielded a high-molecular-weight and a low-molecular-weight fraction. The high-molecular-weight fraction contains mainly carbohydrate (68.7%) and small amount of protein (3.6%), whereas the low-molecular-weight fraction contains 31% carbohydrate and was devoid of protein. Only the high-molecular-weight fraction exhibited antiproliferative activity against cancer cells, with IC(50) of 70.0 μg/mL and 76.7 μg/mL, respectively. Thus, the cytotoxic action of the LR-CW is due to the high-molecular-weight fraction, either the proteins or protein-carbohydrate complex.
Ganoderma lucidum (M.A. Curtis:Fr.) P. Karst is a popular medicinal mushroom. Scientific reports had shown that the wound healing effects of G. lucidum were partly attributed to its rich polysaccharides. However, little attention has been paid to its potential effects on wounds associated with diabetes mellitus. In this study, we evaluated the wound healing activity of the hot aqueous extract of G. lucidum in streptozotocin-induced diabetic rats. The extract of G. lucidum was standardised based on chemical contents (w/w) of total polysaccharides (25.1%), ganoderic acid A (0.45%), and adenosine (0.069%). Six groups of six rats were experimentally wounded in the posterior neck region. Intrasite gel was used as a positive control and aqueous cream as the placebo. Topical application with 10% (w/w) of mushroom extract-incorporated aqueous cream was more effective than that with Intrasite gel in terms of wound closure. The antioxidant activity in serum of rats treated with aqueous extract of G. lucidum was significantly higher; whereas the oxidative protein products and lipid damage were lower when compared to those of the controls. These findings strongly support the beneficial effects of standardised aqueous extract of G. lucidum in accelerating wound healing in streptozotocin-induced diabetic rats.