Culturability, Cultivation Potential, and Element Analysis of the Culinary-Medicinal Cauliflower Mushroom Sparassis latifolia (Agaricomycetes) from Pakistan.

Sparassis Fr. is a recognized nutritionally and therapeutically significant genus of mushrooms. Its species has the potential to meet the nutritious and healthy food needs of the growing population but unfortunately due to their tough, laborious and undefined cultivation conditions, their domestication is very rare and not properly described before. In this study, first time the detailed culturabilty and cultivation potential of a wild S. latifolia species from Pakistan was evaluated. Among the different media used, maximum cultural growth was observed on pine needles extract agar (PEA) medium at 24°C. Cultured strains on PEA medium were used to spawn on wheat, sorghum and barley grains. Sorghum grains at 24°C were found as the best combination for spawn production of this mushroom. Total yield efficacy was investigated on a variety of substrates. A mixed substrate of Morus sawdust and pine sawdust at 20°C showed the optimum yield. Element analysis of wild and cultivated strain was carried out and revealed that this mushroom is enriched with macronutrients. These results showed that S. latifolia has great artificial growth potential. Its domestication can compete with commonly growing mushrooms in nutritional and pharmaceutical attributes.

Appraisal of Antioxidant Potential and Natural Bioactive Compound Determination through HPLC from Two Saprotrophic Amanita (Agaricomycetes) Species from Pakistan.

The present study was carried out to investigate and identify bioactive compounds along with antioxidant capacity, total flavonoids and total phenolic contents from two saprophytic Amanita species, i.e., mushrooms A. manicata (Berk. & Broome) Pegler and A. nana Singer. Antioxidant potential was assessed by DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging and ABTS (2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) assay, total phenolics, and flavonoids. Both mushrooms were found to possess antioxidants and wide range of phenolics and bioactive compounds. There was maximum percent inhibition (83.2 ± 0.120%) on DPPH by A. manicata. However, maximum percent inhibition using ABTS was found to be 79.5 ± 0.251% by A. nana. Similarly, A. nana possesses maximum amount of both flavonoids and phenolics i.e., 0.3473 ± 0.0088 mg/100 g of catechin and 0.097 ± 0.0011 mg/100 g of GAE equivalent, respectively. Both mushrooms exhibited a variety of natural compounds such as P-coumaric acid, M-coumaric acid, benzoic acid, ferulic acid etc. Ferulic acid, chlorogenic acid, and cinnamic acid were also detected in A. manicata. A. manicata exhibited best and greater antiradical potential than A. nana due the presence of excessive natural bioactive compounds. From the reported results, it was revealed that both inedible mushrooms could be a potential source of antioxidants and secondary metabolites and might be used for making novel drugs in future by pharmaceutical industries.

Antioxidant Potential and Stability of Oxidative Sunflower Oil by Using the Ethanolic Extracts of Saprophytic Edible Cultivable Mushrooms from Pakistan.

During this study, ethanol extracts of four edible mushrooms i.e., Macrocybe gigantea, Podaxis pistillaris, Lentinus tigrinus, and Pleurotus cystidiosus were prepared and investigated by TFC, TPC, FRAP, ABTS and DPPH for their antioxidant potential. The extracts were also used to check the oxidative stabilization of sunflower oil for a period of 45 d using standard methods i.e., peroxide value, free fatty acid contents and iodine value. M. gigantea extract showed the maximum flavonoids and phenolic contents and exhibited the maximum antioxidant potential among the four edible mushrooms. BHA was observed to have maximum oil stabilization potential followed by the ethanol extract of M. gigantea of 1000 ppm concentration, BHT and extract of 1000 ppm concentration of P. pistillaris. Thus, it is concluded from this study that ethanolic extracts of these mushrooms can be used as a healthy supplement to stabilize oil by replacing synthetic antioxidants that are harmful to humans.

Domestication of a magic therapeutical wine glass fungus (Podoscypha petalodes) from Pakistan.

The Podoscypha is a recognized therapeutically significant genus of mushrooms. A Podoscypha species under a Morus sp. Was found from the largest man-made forest Changa-Manga, Kasur during the exploration of fungal flora of Pakistan. Detailed morpho-anatomical and phylogenetic analysis identified it as P. petalodes, a part of common native flora of many regions of the world like Pakistan. The culturabilty and cultivation potential of this fungus was assessed for the first time using different media and substrates. Maximum cultural growth was observed on the Compost Extract Agar (CEA) medium at 28 °C. Mycelium of cultured strains on CEA medium was used for the spawn production on wheat, sorghum and barley grains. Cultivation potential in the form of spawn running period, harvesting time duration and yield was investigated on variety of substrates. A mixed substrate of sawdust and tea waste at 28 °C showed the optimum yield. Tea-waste was used as the casing material in all substrates and proved very effective. These results depicted that Podoscypha petalodes possesses the cultivation potential. Its cultivation on large scale can solve the major health concerns of the growing population. It would provide the people easy accessibility of economical, effective and natural medicine throughout the year that restrict in case of only natural production at specific time of the year.

Extrapolating Qualitative Mycochemical Profiling and Therapeutic Potential of Two Mycorrhizal Amanita Species (Agaricomycetes) from Pakistan.

The present study was an attempt to evaluate the antimicrobial and anthelmintic potential of two Amanita species, i.e., A. orsonii and A. glarea, along with their qualitative mycochemical screening. Maceration technique was adopted to make crude extracts in non-polar (petroleum ether and chloroform) and polar (ethanol and distilled water) solvents. Qualitative mycochemical screening revealed the presence of a variety of secondary metabolites like terpenoids, flavonoids, tannins, alkaloids, saponins, and cardiac glycosides. Antimicrobial activities were carried out by using agar well diffusing method against four bacterial and one fungal strain. The antibacterial potential ranged between 4.86 ± 0.088 mm and 34.83 ± 0.166 mm with maximum inhibition zone exhibited by petroleum ether extract against Pseudomonas fluorescens and least potential by distilled water macerate of A. orsonii against the Escherichia coli. The antifungal activity ranged between 14.5 ± 0.288 and 24.76 ± 0.145 mm, with the highest potential provided by chloroform extract of A. orsonii and least capability put forward by A. glarea by petroleum ether extract against Fusarium solanii. Antibiotics and antimycotic discs were used as standard and some of the crude extracts showed bigger zone of inhibition as compared to standard. Different concentrations of ethanolic extracts of both mushrooms were subjected to anthelmintic potential against parasite Haemonchus contortus. All the crude extracts were more potent than standard oxfandazole used. Anthelmintic potential ranged between 9.9 ± 0.057 and 54.93 ± 0.033 minutes for death of parasite, while the paralyzed time ranged between 4.86 ± 0.088 and 24.86 ± 0.088 minutes. From the results obtained it was concluded that both mushrooms can be used as potential source of curative antibacterial, antifungal, and anthelmintic agents against several diseases that might be used in pharmaceutical industries for making medicines and to screen out secondary metabolites in future.

Mycelial Growth, Basidiocarp Production and Mineral Analysis of the Edible Lentinus squarrosulus (Agaricomycetes) from the Subtropical Part of Pakistan.

Lentinus squarrosulus (Mont.) Singer, a wild edible macrofungus, possesses confirmed organoleptic, nutritional, and medicinal attributes. During the exploration of fungal biota of Punjab, Pakistan, L. squarrosulus was collected under the Morus alba from University of the Punjab, Lahore. For the domestication of this wild significant species, its culturability and cultivation potential was assessed by using different synthetic culture media and substrates. Among these different media used, maximum cultural growth was observed on the malt-extract agar (MEA) medium at 35°C followed by the potato-dextrose agar (PDA), compost extract agar (CEA), glucose-peptone agar (GPA), and Sabouraud-dextrose agar (SDA). Cultured strains on MEA medium were used to produce spawning material on wheat, sorghum, and barley grains. Sorghum grains at 35°C were found to be the best combination for spawn production of this fungus. Yield and biological efficiency were investigated on a variety of substrates. A mixed substrate of wheat straw and tea waste at 30°C showed the optimum yield. Tea-waste medium was used as the casing material and proved very effective. Mineral analysis of the wild and cultivated strain was carried out and revealed that both forms were enriched with macronutrients. These results showed that the L. squarrosulus strain from Pakistan possesses good nutrient and growth potential and its domestication can compete with nutritional and medicinal properties of commonly growing mushrooms.

Evaluation of Mycochemical Analysis and In Vitro Biological Activities of Some Russula Species (Agaricomycetes) from Pakistan.

In this study, mycochemical screening was performed to estimate the in vitro biological activities and antimicrobial and antioxidant potential of Russula aeruginea and R. brevipes. Crude extracts of these mushroom species were obtained by using maceration techniques. Qualitative mycochemical analysis showed the presence of a variety of secondary metabolites such as terpenoids, alkaloids, flavonoids, reducing sugars, and anthraquinones. Antimicrobial activity of crude extracts was assessed by using agar well diffusion methods against virulent bacterial and fungal strains. Ethanolic extract of R. brevipes displayed maximum antibacterial activity, with a 28.4 ± 0.953 mm zone of inhibition against Bacillus subtilis. Likewise, ethanolic extract of R. aeruginea exhibited enhanced antifungal activity, with a 24.6 ± 0.333 mm zone of inhibition against Fusarium equiseti. The antioxidant potential of the ethanolic extracts was assessed using five assays: 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power assay (FRAP), total flavonoid content (TFC), and total phenolic content (TPC). The maximum percent inhibition for R. aeruginea was 87.069 ± 0.007 mg/mL on ABTS radicals. In contrast, the maximum percent inhibition for R. brevipes was 42.166% ± 0.023% and 0.571 ± 0.005 mmol/L of FeSO4 using the DPPH and FRAP methods, respectively. TFC was the same for both mushrooms (0.113 ± 0.002 mg/100 g of catechin), and the maximum TPC was 0.066 ± 0.00 mg/100 g of gallic acid equivalents. From these results, it was concluded that R. aeruginea and R. brevipes could be a source of new antimicrobial and antioxidant agents and may be useful for the development of novel drugs in the future.

Mycochemical Screening and Study of Medicinal Properties of Two Amanita Species (Agaricomycetes) from Pakistan.

Fresh basidiocarps of Amanita cinnamomescens and A. pakistanica were collected from Ayubia-Khanspur, Pakistan, during the 2018 monsoon season. Basidiocarps of A. cinnamomescens and A. pakistanica were evaluated for their mycochemicals, mineral composition, and antioxidant and antimicrobial activities. The percentage yield of extracts ranged from 4.13% to 18.20%. The extracts contained noticeable total phenolic contents (0.043 ± 0.02 to 0.046 ± 0.01 mg/g) and total flavonoid contents (0.090 ± 0.004 to 0.0935 ± 0.003 mg/g) and good radical scavenging ability according to the ABTS assay (79.74% ± 0.03% to 85.34% ± 0.02%) and the DPPH radical assay (35.77% ± 0.01% to 44.77% ± 0.003%). In addition, the tested extracts showed substantial antimicrobial activity, which ranged from 10 ± 0.33 to 32.66 ± 0.33 mm. Both mushrooms were also analyzed for their mineral content (sodium, potassium, calcium, nickel, cobalt, copper, zinc, manganese, chromium, and iron). It was concluded that A. cinnamomescens and A. pakistanica can be used as a potential source for formulation of dietary functional foods and pharmaceuticals with antioxidant and antimicrobial effects. To our knowledge, this is the first report on in vitro biological activities and mycochemical analysis of A. pakistanica and A. cinnamomescens from Pakistan.

Proximate Analysis and In Vitro Biological Activities of Cauliflower Mushroom, Sparassis crispa (Agaricomycetes), from Pakistan.

This study evaluates the proximate analysis and antibacterial and antioxidant potential of the edible mushroom, Sparassis crispa. The preliminary mycochemical analysis revealed the presence of secondary metabolites such as saponins, terpenoids, flavonoids, tannins, and cardiac glycosides. Proximate analysis was performed to estimate the presence of some essential and nonessential metals in the sample. Among the minerals analyzed, zinc was in the maximum range (1.156 mg/g) compared to the other elements. The antioxidant potential of S. crispa ethanolic extract was assessed by using five assays: 1) 2,2'-azino-bis-3-ethylbenzothiazoline-6-suphonic acid, 2) ferric reducing antioxidant power, 3) 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), 4) total flavonoid content, and 5) total phenolic content. The DPPH radical scavenging assay confirmed the highest percent inhibition of the extract (56.43% ± 0.21%). Antibacterial activity of the mushroom sample was tested against the selected Gram-negative and Gram-positive bacterial strains by using the agar well diffusion method. S. crispa ethanolic extract exhibited maximum antibacterial activity with an inhibition zone of 19.66 ± 0.88 mm against Escherichia coli compared to other bacterial strains. From these results, it could be assessed that S. crispa is a promising source of new antibacterial and antioxidant agents.