INTRODUCTION:

Mushrooms are an essential product of forest ecosystems that grow on the most abundant nutrients, like cellulose. A mushroom is a macro-fungus that has a distinctive fruiting body. It is suggested that only 2000 are safe for edible to human beings, and about 650 of these possess medicinal value out of approximately 15,000 known species in the world¹. Mushrooms have been chiefly used as a portion of human food for centuries and have been famous for their texture, flavour, and medicinal properties.

However, mushrooms recently emerged as an essential source of biologically active material with therapeutic value. Wild-growing mushrooms contain several secondary metabolites, carbohydrates, minerals, proteins, fibres, vitamins and fats with antitumor, antifungal, antimicrobial and antioxidant properties^2,38,40.

It is suggested that mushrooms are also essential sources of compounds like beta-glucans, ascorbic acid, tocopherols, carboxylic acids, lectins, terpenoids and various dietary fibres^3,39.

The commercial production of mushrooms is growing worldwide, producing approximately more than two million tons annually⁴. Mushrooms are a poor source of amino acids like sulfur-containing methionine and cysteine but rich in beneficial minerals such as iron, copper, zinc, and manganese, essential for biological systems. Nowadays, mushrooms have become famous as a source for the development of drugs from a natural source, nutraceuticals, as well as functional food. Medicinal mushrooms are considered treatments for many diseases but remain unconfirmed in mainstream science and medicine, therefore not approved as a drug or medical treatment. Previous research has shown that some medicinally valued mushrooms are used for treating human diseases such as cardiovascular, anticancer, antiviral, antibacterial, antiparasitic, anti-inflammatory, and antidiabetic properties^{5,41,44,47,48}. Therefore, the mushroom is very interested in using dietary supplements because they contain a massive category of biological components. Mushrooms are a non-animal-based food containing vitamins for vegetarians⁶. The bioactive compounds in mushrooms capable of scavenging free radicals to protect living organisms from oxidative damage also play an essential role in defending and curing unwanted physiological effects. Therefore, scientists show interest in developing dietary materials from natural sources to consumer preference. Mushrooms have become famous as nutritionally beneficial foods and a drug development source⁷. Also, extracted polysaccharides in edible mushrooms are one of the most critical components correlated with multiple bioactive compounds, such as antioxidant, reducing blood lipid, antitumor and antimicrobial activities, etc.^8,42,43. The present study selected S. commune, which belongs to basidiomycete, and completed its life cycle in 8- 10 days. The ecotype nature is saprobic on dead wood, which grows alone or, more frequently, in clustered ways⁹. Schizophyllan is a critical bioactive content produced from S. commune; they are homopolysaccharide, which consists of a linear chain of β-d-(1-3)-glucopyranose groups and β-d-(1-6)-glucopyranosyl group, have homopolysaccharide, and non-ionic and water-soluble which is consisting of a linear chain of β-d-(1-3)-glucopyranose groups and β-d-(1-6)-glucopyranosyl groups produced by S. commune. Similarly, Daedaleopsis confragosa belongs to the family of Polyporaceae, which is a highly variable species; the presence of maze-like pores, commonly whitish to brownish cap with a zone of colour and its pore surface to bruise reddish, which is found in hardwood and stumps¹¹. Laetiporussulphureus (Bull.) Murrill belongs to a class of Aphyllophorales. It has widely dispersed in Asia, Europe and North America. It is commonly found on hardwood-dried tree trunks. The fruiting bodies are substantial and extend beyond to others, their form as bunches in 6–45. It has a leathery, spongy appearance and commonly appears bright or light orange and its lifespan is annual¹¹. This present study intended to investigate extract preparation using various polar solvents and then determine the in vitro antimicrobial capacity of the extract using minimum inhibitory concentration.

MATERIALS AND METHODS:

Sample Collection:

Fresh, dried polypore mushroom samples were collected from Charama forest, Kanker district and Thakurtola forest, Rajnandgaon district (Chhattisgarh). These collected samples were kept in polythene bags, packed loosely, and given a specific code. Identification was made based on critical observations of the specimens and examination of relevant literature^9,12,45. The D. confragosa sample, which was collected from the Charama forest region, was given to code (C); the D.confragosa sample, collected from Thakurtola forest, code to (F), L. sulphureus, obtained from Charama forest, given to (TS) and S.commune, was obtained from Charama forest, code to (SC) respectively. All the identified and unidentified mushroom samples were deposited to Dept. School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, for further analysis. All of the chemicals and other reagents were purchased from HiMedia Pvt. Ltd. Mumbai and Sigma Aldrich Bangalore, India.

Microorganisms Used:

In the present study, human and non-human pathogenic microorganisms were taken to assay the antimicrobial efficiency of polypore mushrooms' water, methanol and ethanol extracts. Microorganisms showed various pathogenicity properties for human beings like B. cereus McR-3 is (human pathogenic), B. subtilis BAB 2437 (non-human pathogen), K. oxytoca ATCC 13182 (human pathogenic), and fungus-like A. ustus MTCC-2200 (human pathogenic) and C. elegans MTCC 552 (non-human pathogen) nature, were used as an assessment of antimicrobial activities of mushroom extracts. Bacteria cultures were stored in slants containing nutrient agar medium (NAM), and fungal cultures were stored on potato dextrose agar (PDA), respectively. The bacterial suspension was enriched in NAM for 24 hours, fungi on PDA for 72 hours, and 1-1.5x106 cells/ml fresh inoculums were utilized for antimicrobial assay^2,13,48.

Extraction Methods:

a. Water:

This procedure was carried out according to method ¹⁴ with slight modification. Mushrooms were cut into small pieces, and then 25g of mushrooms were employed in a water bath at 100^◦C for 2 hrs after the water extract was filtered through Whatman filter paper number 1. The extracts of various mushroom samples were denoted to a particular code, shown in (Table 1).

Methanol:

Methanol extraction of mushrooms was done using the method¹⁵. All dried mushrooms were crushed with the help of a motor pestle to make a fine powder. 25g of fine powder was stirred in a shaking incubator with 250ml of methanol at 25⁰C at 150rpm for 24 hours and filtered through man filter paper number 1. Then, methanolic extract evaporated at 40⁰C for dryness. The extracts of various mushroom samples are denoted to a particular code, shown in (Table 1).

Ethanol:

According to methodology, the procedure was done with slight modification¹⁵; 25g of mushroom powder was stirred in a shaking incubator with 250ml of 99.9% ethanol at 25⁰C at 150rpm for 24hours. After being centrifuged at 3000 rpm for 15 min., the sample was filtered by man filter paper number 1. Further, ethanolic extract was evaporated at 40⁰C for dryness. The extracts of various mushroom samples are denoted to a particular code, shown in (Table 1). All the extracts of mushroom samples (including water, methanol and ethanol solvent medium) were dissolved in 75% DMSO solution and kept at 4⁰C.

Qualitative Screening of Bioactive Contents of Polypore Mushroom:

Potential qualitative bioactive contents screening according to methodology^16,17with slight modification, displayed in table 2. These were the following:

1. Reducing sugar (Fehling's Test):

5ml aqueous ethanol extract was taken in a test tube, and a few drops of Fehling's solution A and B were added, and the colour reaction showed the presence of reducing sugar.

2. Terpenoids (Salkowski Test):

The 5ml extract was added with 2ml chloroform and 3 ml concentrated sulphuric acid to form a layer. The brownish colour showed the presence of terpenoids.

3. Flavonoids Test:

Small quantities of filtrate extract were added with 5ml of dilute ammonia and 1ml of concentrated sulphuric acid. The appearance of yellow colour indicated the presence of flavonoids.

4. Saponins Test:

5 ml of water extract was shaken, and when it formed stable, persistent added three drops of olive oil, then again shacked vigorously. The formation of emulsion showed a positive result.

5. Tannins Test:

10 ml water extract was boiled and then filtered the solution. A few drops of 1% ferric chloride solution were added to the filtrate and observed for brownish green or blue-black precipitate.

6. Cardiac glycosides (Keller-Killiani Test):

2 ml glacial acetic acid, one drop ferric chloride and 1 ml concentrated sulphuric acid were taken and added to 5 ml of water extract. The formation of a brownish ring indicated a positive result.

7. Anthraquinones Test:

After filtering the solution, 5ml extract was boiled with 10 ml concentrated sulphuric acid. 5 ml chloroform was added to the filter; the chloroform layer was pipetted to another test tube, and 1ml was diluted with ammonia solution. Changes in the colour of the resolution showed the presence of anthraquinone.

8. Phlobatannins:

The water extract of the mushroom was boiled with 1% hydrochloric acid. The deposition of red precipitation indicated the presence of phlobatannins.

9. Carotenoids:

1 ml mushroom extract was shaken vigorously with 10 ml chloroform and filtered. After that, 85% concentrated sulphuric acid was added to the filter. The formation of a blue colour showed the presence of carotenoids.

10. Phenols (Ferric chloride test):

1 ml of ethanolic extract from the sample was added to 2 ml distilled water; a few drops of 10% ferric chloride were added. The appearance of blue or green colour indicated the presence of phenol.

Antimicrobial Assay:

The antimicrobial potency was analyzed according to the procedure with slight modifications^13,46. The typed microorganism's culture was obtained from the stock collection centre, Department of School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur (CG) India. The following strains of bacteria were used for the antibacterial assay: B. cereus McR-3, B. subtilis BAB 2437, and K. oxytoca ATCC 13182. All pathogenic bacteria were inoculated in nutrient broth medium at 35±2⁰C for 24 hours, used for testing the antibacterial activity of mushrooms, and the fungus strains A. ustus MTCC-2200 and C. elegans MTCC 552 were inoculated in potato dextrose broth medium and incubated at 25±2⁰C for 24 to 72hours. Antimicrobial activity was performed using the micro-dilution method using a microtitre plate. Initially, different concentration of the mushroom extract was poured into a microtitre plate and then made with autoclaved distilled water up to 200µl, then 100 sterile nutrient broth for bacteria and potato dextrose broth to provide a support medium, after adding 50µl of culture and bacterial plates were incubated for 24hrs. at 35±2⁰C and fungal plates were incubated for 48 to 72hours, at 25±2⁰C. After incubation, an ELISA reader determined absorbance at 600nm and 595nm sequentially.

Statistical analysis:

All the experiments were performed with three parallel measurements. Data were analyzed by using one-way variance analysis (SPSS 16.00). The significant levels were defined at P<0.05. SigmaPlot 12.0 created the graphical 2-D data. The results are here presented as mean values ±standard error (SE).

RESULTS:

In the present study, some specific polypore mushrooms have been collected from the mixed dense forest of Charama, located in District Kanker and other samples procured from Thakurtola forest, which belongs toRajnandgaon district, Chhattisgarh. The collected mushroom was morphologically identified using the standard procedure. There were three different polyporeace mushrooms collected. Among them, D. confragosa was one of two distinct geographical regions of Chhattisgarh. D.confragosa, which has characterized morphology with characteristic studies with available monographs and related literature, is called a blushing bracket because it has pink or purple shades on the upper surface and is a low-growing fungus. It is mainly found in temperate regions. Because of their hardness and woody structure, they are usually inedible, except for a few particular edible species with medicinal values¹⁸. Both collected samples of species were similar. S. commune was a whitish grey and had fan-shaped gilled fruit bodies. Its season of fruiting is a perennial, and it lives on living and dead hardwood trees. L. sulphureus is a multiple cluster of yellow-orange shelves growing on wood; it is very soft, fleshy when young and turning hard when mature. It's generally grown in summer and fall. It causes a brown cubical rot of living and dead hardwood and conifer trees. When it is in its early stage, this mushroom is usually edible. Macro fungi and human beings have intimate biological relationships in traditional medicinal systems. Many such polypore fungi are commonly used for potential therapeutic value product development.

Yield of Mushroom Extracts:

Based on the morphological appearance characterized, they were subjected to various solvents for extraction, as shown in Table 1. In the present study, three solvents, Hot Water [100⁰C], Methanol and Ethanol, were used in the extraction process.

D. confragosa, collected from the Charama forest, was the best-extracting solvent among the extracted medium water, with a percentage recovery of 50.00. This was followed by ethanol with a percentage recovery of 3.65, while methanol extracted only 2.7g/L. Similarly, D. confragosa was obtained from the Thakurtola forest; hot water extracted solvent was given 4.5 percentage of recovery, followed by ethanol with a yielding percentage of 1.50 and the lowest yield rate recorded for methanol, i.e. 0.59, respectively. For S. commune, hot water extract had the highest recovery percentage of 19.6, followed by methanol and ethanol, with a 14.00 recovery percentage recorded. In the same way, L. sulphurous, in methanol extract medium, gave the highest recovery percentage, i.e. 11.1, hot water had 9.8, and the lowest recovery percentage (8.3) was recorded for ethanol extraction medium, respectively. The extract solvent system is based on polarity, giving the diverse nature of efficient yields. Hence, selecting solvent and the defatted procedure is vital for obtaining qualitative and quantitative myco contents.

Phytochemical Screening:

The phytochemical screening of mushroom extracts is shown in table number 6. According to the present study, D. confragosa (CH) was collected from the Charama forest, and various polar solvents were extracted and analyzed phytochemical screened; a test revealed that the moderate amount of reducing sugar, terpenoids, flavonoids, saponins, tannins and other constituents showed in trace amounts such as Cardiac glycosides, Anthraquinones and Carotenoids. Similarly, the D. confragosa (F) from Thakurtola forest samples has shown a high amount of terpenoids and flavonoids, and other constituents such as reducing sugar, saponins, tannins, and cardiac glycosides were observed at moderate levels. The remaining bioactive contents, such as anthraquinones, phlobatamines and carotenoids, were the least and showed trace or reasonable amount; as compared to CH, F bioactive contents have less potent might be caused by geographical origin regions have varied qualitative phytochemical assayed. S. commune (SC) showed only saponins, tannins, and anthraquinones in trace amounts, whereas other constituents showed fewer amounts, and carotenoids were absent in this sample.

Among all samples of L. sulphureus (TS), a high amount of all constituents' only phlobatannins and carotenoids were present in moderate, and anthraquinones were present in fewer amounts. Geographically, regions also affected the nature of bioactive contents; these studies suggested that the D. confragosa polyporeace mushroom is a good alternative for medical application. The Thakurtola forest region selected more vigorous mushrooms due to their potent antimicrobial properties. The present study suggested that the constituents of mushrooms showed an efficient antimicrobial activity.

Antimicrobial Assay:

The antimicrobial, including antibacterial and antifungal activity, was determined by microdilution method using 96 wells micro-titre plate. The minimum inhibitory concentration (MIC) was the lowest; there was no macroscopically visible growth in the in vitro system. The microbial growth was recorded in the turbidity and presence of the pellet at the bottom of the well. Different concentrations of mushroom extracts were initially poured into the microtitre plate; after the incubation periods, microbial density was observed, and the MIC value of both antifungal and antibacterial activity of all mushroom extracts was represented in Tables 3, respectively.

The present study observed that the mushroom extracts from different solvents showed potent antimicrobial activity against human pathogenic and non-pathogenic microbes; it has recorded that the methanolic extracts of h D. confragosa from Charamaforest that the quantity of mushroom extract 20µg/µl concentration has minimal concentration for (MIC) against fungus A. ustus MTCC 2200 and C.ellaelegans MTCC 552, whereas D. confragosa from Charama forest has least potent, was 120µg/µl MIC against C. elegans MTCC 552. Similarly, the hot water and ethanolic extracts revealed the dose repose assayed of antifungal activity.

Table 3. Minimum inhibitory concentration (MIC) of extract of polypore mushrooms against pathogenic fungi.

SN	Mushroom	Extraction solvent	Antifungal activity MIC (µg/µl)
SN	Mushroom	Extraction solvent	*A. ustus* MTCC 2200	*C. elegans* MTCC 552
1	CHE	Water	20.00	20.00
2	FHE		20.00	100.00
3	SCHE		20.00	40.00
4	TSHE		20.00	20.00
5	CME	Methanol	40.00	120.00
6	FME		20.00	20.00
7	SCME		20.00	20.00
8	TSME		20.00	20.00
9	CEE	Ethanol	40.00	40.00
10	FEE		20.00	20.00
11	SCEE		20.00	100.00
12	TSEE		20.00	20.00

Growth inhibition was observed based on increasing doses with decreased growth of fungal biomass. Similarly, water extract of mushroom contents revealed antimicrobial activity, and it was observed that the Charama forest collected D. confragosa as a potent antimicrobial, compared to Thakurtola collected mushroom. Similarly, L. sulphureus has the 20µg/µl MIC against bacteria B. cereus McR-3 and K.oxytoca ATCC 13182. Antibacterial activity of water extracts from several polypore mushrooms against B.subtilis BAB 2437have characterized; the graph depicted that increasing concentration of extracts has shown potent antimicrobial activity. The highest growth inhibition was observed at120 µg/µl and the lowest growth inhibition was recorded at an initial concentration of 20 µg/µl. The diagrammatic presentation is shown in figure 1.

Figure 1. Antibacterial activity of water extracts from several polypore mushrooms against B.subtilis BAB 2437. Each value is expressed as mean ± standard error, with % inhibition of bacterial growth (n=3)

The antibacterial activity of methanolic extracts from several polypore mushrooms against B. subtilis BAB 2437 has been assayed; the graph depicts that the gradual increase in the concentration of extracts observed has shown potent antimicrobial activity. The highest growth inhibition was observed as follows in 120 µg/µl, and the lowest growth inhibition was recorded at an initial concentration of 20 µg/µl. Among the polyporeace mushroom species, S. commune and D. confragosa Thakurtola forest samples have potent antibacterial and other extracts have less powerful activity. Antibacterial activity against K.oxytocaATCC 13182 was recorded for different extracts. Ethanolic, methanolic and hot water extracts have shown potent activity with correlated on when dose increasing of extracts, microbial growths have declined; the graph depicted that gradually increasing the concentration of extracts progressively microbial growth has inhibited. The highest growth inhibition was observed as followed in 120 µg/µl, and the lowest growth inhibition was recorded at an initial concentration of 20 µg/µl. Among the polyporaceae mushroom species S. commune and D. confragosa, Thakurtola forest samples have potent antibacterial and other extracts such as L. sulphurous have less potent activity.

On the other hand, S. commune ethanolic extracts have shown potent antimicrobial properties, and L. sulphureus methanolic extracts (TSME) showed 120µg/µl MIC against K. oxytoca ATCC 13182. Similarly, S. commune (SCME) methanolic extract has shown less potent antibacterial activity against tested pathogenic microorganisms. The present study revealed that the polyporeace mushroom has significant potential for antimicrobial alternative bio resources, an opportunity for a new dimension for drug discovery.

DISCUSSION:

Macro fungi and human beings have had intimate biological relationships since ancient times. Mushroom has a potential medicinal value product; thus, the medicinal mushroom has long been an essential part of human life. The possible nutritive values or therapeutic properties of polypore mushrooms have recently been attracted to worldwide scientific communities for endearing research. The macrofungi are different from other fungi; they have spore-forming fruiting body structures that we know as mushrooms. They have been utilized for many years in Asian countries as a portion of very nourishing food and medicine. The highest recovery value for G. resinaceum in a methanol-ethanol combination extraction medium was recorded at 8.10, followed by ethanol at 7.85, and methanol gave the lowest recovery percentage, i.e., 6.40. They also reported that saponin was present, while anthraquinones were not in the trace amount level in the wild mushroom sample (Ganoderma species)¹⁴. Liew et al.¹⁵performed a phytochemical screening of seven species of wild mushrooms. They reported that triterpene and steroids were completely absent while recording only the highest saponins in L. sajor-caju. The phytochemical screening evaluated Ganoderma sp. and recorded active medicinal constituents, such as saponins, terpenoids, phlorotannins, tannins, etc. They suggested that active compounds are mainly responsible for their medicinal properties²⁰.

Further, the most outstanding amount of phyto-metabolites showed higher values for antioxidant, antimicrobial, and essential parts of drug discovery. They found 5mg/ml & 2mg/ml MIC against A. niger & B. subtilis of ethanolic extract of Momordica cymbalaria found that 22.6 µg/µl, 22.4 µg/µl and 33.720µg/µl MIC from ethanol, methanol and water extracts of Melia azedarachi against B.subtilis. They further record that the MIC of M. azedarachi from ethanol, methanol and water extracts for 51.6 µg/µl, 49.6 µg/µl and 55.2µg/µl respectively. This finding is similar to our present experimentation finding¹³. Simillilary worked on Agaricus bisporus and evaluated the antimicrobial activity of essential oils and their components against the three major pathogens of the cultivated button mushroom²¹. They reported antimicrobial properties of the ethanolic extract from Lepista nuda (Bull.)²². Another researcher⁷ worked on antimicrobial activity against some microbes and bioactive compounds of Portuguese wild edible mushrooms (Lactarius deliciosus, Sarcodon imbricatus and Tricholoma portentosum) methanolic extracts. According to the²³, organic solvents and supercritical fluids obtain the antioxidant and antimicrobial activities of shiitake (L.edodes) extracts. Theystudied²⁴ the antimicrobial and antineoplastic activity of P. ostreatus. The antimicrobial activity of Melaleuca alternifolia essential oil on an antagonistic potential of Pleurotus species against Trichoderma harzianum in dual culture²⁵.

Similarly, they worked on free radical scavenging and antimicrobial properties of extracts of wild mushrooms of T.clypeatus, T. srobustus, L. subnudus and Lenzites species²⁶. He suggested the antimicrobial properties of shiitake mushrooms (L.edodes)²⁷. They studied antioxidant and antimicrobial activity; they found that a good amount of total phenolic content reveals the potency of antimicrobial and potency of antioxidants of polypore family mushrooms. The present research showed that the bioactive and their significant contents indicated the biological effectivity²⁸. According to the³⁰, they extracted concentrate and successfully analyzed the antimicrobial and antioxidant activities of Russula delica. They studied the effects of gamma irradiation on the chemical composition and antioxidant activity of Lactarius deliciosus L. wild edible mushrooms³⁰.They reported nutritional value, chemical composition, antioxidant activity, and enrichment of cream cheese with chestnut mushroom Agrocybe aegerita (Brig.) Sing¹. They efficaciously screened the antimicrobial, antioxidant properties and bioactive compounds of edible mushrooms of P.ostreatus, L.edodes and Hypsizigus tessulatus cultivated in Bangladesh². They deliberated the function neutraceutical properties of the methanolic extract and effectively analyzed the antimicrobial properties of extracts^31,32. They evaluated the metal concentration and antioxidant, antimicrobial and anticancer potentials of two edible mushrooms L.deliciosus and Macrolepiota procera³³.

Similarly, the antibacterial and cytotoxic activities of wild mushroom Fomes fomentarius (L.) were studied³⁴. They evaluated the antimicrobial properties of different mushroom species of Phaeolus schweinitzii, Inonotus hispidus, Tricholoma columbetta, T. acaligatum, Xerocomus chrysenteron, Hydnellum ferrugineum, A. bosporus and P. ostreatus³⁵. Likely, work done³⁶ and evaluated the chemical constituents and antibacterial properties of medicinal mushrooms G. applanatum, G. lipsiense, G. chalceum and G. tsugae collected from similar. They studied the antibacterial activities of sulphated polysaccharides from P. eryngii and Streptococcus thermophilus ASCC 1275³⁷. According to the researcher²⁰, phytochemical screening quality directly correlates with the biological activity of medicinal mushrooms of some Ganoderma species. The interpretation is the same as that found in our experiment. The current study suggested that the taken polyporace family of mushrooms has a broad bioconstituetns category of active contents that would be good resources for nutritional production and drug development; it has also provided a prominent option for immune-modulatory related dietary supplements and antimicrobial drug agents.

CONCLUSION:

In the present work, we collected polypore macro fungus from the forest of Chhattisgarh and successfully analyzed phytochemical screening and potent antimicrobial. The mushroom extracts were prepared with different polar solvents, including methanol, ethanol, hot water, etc. The macerated extracts were successively collected and analyzed for their qualitative various bioactive components; all possible contests were successfully screened out. The antimicrobial assay was characterized by minimum inhibitory concentration (MIC), and minimum dose was described for individual solvent extracts mushroom contents. The study suggested selecting a suitable solvent medium for performing the extraction procedure and choosing the antimicrobial assay. The studies recommend that the D. confragosa, S. commune and L.sulphureus have enormous medicinal and nutritional values, which will be used for various human diseases and provide substitute sources for the pharmaceutical valued product development.

ACKNOWLEDGMENT:

The authors sincerely thank the Head of the Department, School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur (Chhattisgarh). The author also sincerely thanks the Chhattisgarh Council of Science and Technology, Raipur Chhattisgarh, India, for providing financial assistance (Endt. No. 722/CCOST) to successfully execute the studies.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

REFERENCES:

1. Petrovic J, Glamoclija J, Stojkovic D, Ćiric A, Barros L, Ferreira IC, Sokovic M. Nutritional value, chemical composition, antioxidant activity and enrichment of cream cheese with chestnut mushroom Agrocybe aegerita (Brig.) Sing. Journal of Food Science and Technology. 2015; 52: 6711-8.

2. Chowdhury MM, Kubra K, Ahmed SR. Screening of antimicrobial, antioxidant properties and bioactive compounds of some edible mushrooms cultivated in Bangladesh. Annals of Clinical Microbiology and Antimicrobials. 2015; 14(1): 1-6.

3. Babu DR, Rao GN. Antioxidant properties and electrochemical behavior of cultivated commercial Indian edible mushrooms. Journal of Food Science and Technology. 2013; 50: 301-8.

4. Gessler, N. N., Aver, A. A. & Belozerskaya, T. A. Reactive oxygen species in regulation of fungal development. Biochemistry (Moscow), 2007; 72(10): 1091-1109.

5. Cheung, L. M. & Cheung, P. C. K. Mushroom extracts with antioxidant activity against lipid peroxidation. Food Chemistry, 2005; 89: 403–409.

6. Jayakumar T, Thomas PA, Sheu JR, Geraldine P. In-vitro and in-vivo antioxidant effects of the oyster mushroom Pleurotus ostreatus. Food Research International. 2011; 44(4): 851-61.

7. Barros L, Calhelha RC, Vaz JA, Ferreira IC, Baptista P, Estevinho LM. Antimicrobial activity and bioactive compounds of Portuguese wild edible mushrooms methanolic extracts. European Food Research and Technology. 2007; 225: 151-6.

8. Liu J, Jia L, Kan J, Jin CH. In vitro and in vivo antioxidant activity of ethanolic extract of white button mushroom (Agaricus bisporus). Food and chemical toxicology. 2013; Jan 1; 51: 310-6.

9. Kuo, M., Daedaleopsis confragosa. Mushroom Expert.Com, 2005 Website:http://www.mushroomexpert.com/daedaleopsis_confragosa.html,12.21 PM, Date 24/06/2016.

10. Chandrawanshi NK, Tandia DK, Jadhav SK. Nutraceutical properties evaluation of Schizophyllum commune. Indian J. Sci. Res. 2017; 13(2): 57-62.

11. PetrovicJ, Glamoclija J, Stojkovic DS, Ciric A, Nikolic M, Bukvicki D, Guerzoni ME, Sokovic MD. Laetiporus sulphureus, edible mushroom from Serbia: Investigation on volatile compounds, in vitro antimicrobial activity and in situ control of Aspergillus flavus in tomato paste. Food and Chemical Toxicology. 2013; 59: 297-302.

12. Kuo, M. Agaricus arvensis: The horse mushroom. Retrieved from the Mushroom Expert. Com Web, 2007, http://www.mushroomexpert.com/agaricus_arvensis.html

13. Balkhande SV, Surwase BS. Antimicrobial activity of tuberous root extracts of Momordica cymbalaria hook. Asian J. Pharm. Clin. Res. 2013; 6(1): 201-3.

14. Akinyele BJ, Obameso JO, Oladunmoye MK. Phytochemical screening and antimicrobial potentials of three indigenous wild Ganoderma mushrooms from Ondo state, Nigeria. Nigerian Journal of Microbiology. 2011; 25: 2280-90.

15. Elmastas M, Isildak O, Turkekul I, Temur N. Determination of antioxidant activity and antioxidant compounds in wild edible mushrooms. Journal of Food Composition and Analysis. 2007; 20(3-4): 337-45.

16. Sofowora, A. Medicinal plants and Traditional Medicine in Africa; Spectrum Books, Ibadan. 150 Trease, G. E. and Evans, W.C. (1989) Pharmacognosy; (13th edition) Bailliere Tindall, London, 1993 176-180.

17. Trease, G. E. and Evans, W.C. Pharmacognosy; (13thedition) Bailliere Tindall, London, 1989, 176-180.

18. Vidovic, S., Zekovic, Z., Mujic, I., Lepojevic, Z., Radojkovic, M. & Zivkovic, J. The antioxidant properties of polypore mushroom Daedaleopsis confragosa. Central European Journal of Biology. 2011; 6(4): 575–582.

19. Liew GM, Khong HY, Kutoi CJ, Sayok AK. Phytochemical screening, antimicrobial and antioxidant activities of selected fungi from Mount Singai, Sarawak, Malaysia. Int J Res Stud Biosci. 2015; 3(1): 191-7.

20. Kandhasamy, R. Dharumadurai, D. Phytochemical screening and biological activity of medicinal mushroom Ganoderma species. Malaya Journal of Biosciences. 2014; 1(2): 67-75.

21. Sokovic M, Van Griensven LJ. Antimicrobial activity of essential oils and their components against the three major pathogens of the cultivated button mushroom, Agaricus bisporus. European Journal of Plant Pathology. 2006; Nov; 116: 211-24.

22. Mercan N, Duru ME, Turkoglu A, Gezer K, Kivrak I, Turkoglu H. Antioxidant and antimicrobial properties of ethanolic extract from Lepista nuda (Bull.) Cooke. Annals of Microbiology. 2006; 56: 339-44.

23. Kitzberger CS, Smânia Jr A, Pedrosa RC, Ferreira SR. Antioxidant and antimicrobial activities of shiitake (Lentinula edodes) extracts obtained by organic solvents and supercritical fluids. Journal of Food Engineering. 2007; 80(2): 631-8.

24. Wolff ER, Wisbeck E, Silveira ML, Gern RM, Pinho MS, Furlan SA. Antimicrobial and antineoplasic activity of Pleurotus ostreatus. Applied Biochemistry and Biotechnology. 2008; 151(2-3): 402-12.

25. Angelini P, Pagiotti R, Granetti B. Effect of antimicrobial activity of Melaleuca alternifolia essential oil on antagonistic potential of Pleurotus species against Trichoderma harzianum in dual culture. World Journal of Microbiology and Biotechnology. 2008; 24: 197-202.

26. Oyetayo VO. Free radical scavenging and antimicrobial properties of extracts of wild mushrooms. Brazilian Journal of Microbiology. 2009; 40: 380-6.

27. Ps, S. Antimicrobial properties of shiitake mushrooms (Lentinula edodes). International Journal of Antimicrobial Agents, 2009; 33: 591–592.

28. Chandrawanshi NK, Tandia DK, Jadhav SK. Determination of antioxidant and antidiabetic activities of polar solvent extracts of Daedaleopsis confragosa (Bolton) J. Schrot. Research Journal of Pharmacy and Technology. 2018; 11(12): 5623-30.

29. Yaltirak T, Aslim B, Ozturk S, Alli H. Antimicrobial and antioxidant activities of Russula delica Fr. Food and Chemical Toxicology. 2009; 47(8): 2052-6.

30. Fernandes A, Antonio AL, Barreira JC, Botelho ML, Oliveira MB, Martins A, Ferreira IC. Effects of gamma irradiation on the chemical composition and antioxidant activity of Lactarius deliciosus L. wild edible mushroom. Food and Bioprocess Technology. 2013; 6: 2895-903.

31. Kozarski M, Klaus A, Vunduk J, Zizak Z, Niksic M, Jakovljevic D, Vrvic MM, Van Griensven LJ. Nutraceutical properties of the methanolic extract of edible mushroom Cantharellus cibarius (Fries): Primary mechanisms. Food and Function. 2015; 6(6): 1875-86.

32. Chun S, Gopal J, Muthu M. Antioxidant Activity of Mushroom Extracts/Polysaccharides-Their Antiviral Properties and Plausible AntiCOVID-19 Properties. Antioxidants (Basel). 2021; 10(12): 1899. doi: 10.3390/antiox10121899.

33. Kosanic M, Rankovic B, Rancic A, Stanojkovic T. Evaluation of metal concentration and antioxidant, antimicrobial, and anticancer potentials of two edible mushrooms Lactarius deliciosus and Macrolepiota procera. Journal of Food and Drug Analysis. 2016; 24(3): 477-84.

34. Kolundzic M, Grozdanic NĐ, Dodevska M, Milenkovic M, Sisto F, Miani A, Farronato G, Kundakovic T. Antibacterial and cytotoxic activities of wild mushroom Fomes fomentarius (L.) Fr., Polyporaceae. Industrial Crops and Products. 2016; 79: 110-5.

35. Smolskaitė L, Venskutonis PR, Talou T. Comprehensive evaluation of antioxidant and antimicrobial properties of different mushroom species. LWT-Food Science and Technology. 2015; 60(1): 462-71.

36. Singdevsachan SK, Patra JK, Tayung K, Thatoi H. Chemical constituents, antioxidative and antibacterial properties of medicinal mushrooms collected from Similipal Biosphere Reserve, Odisha, India. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. 2017; 87: 559-70.

37. Li S, Shah NP. Antioxidant and antibacterial activities of sulphated polysaccharides from Pleurotus eryngii and Streptococcus thermophilus ASCC 1275. Food Chemistry. 2014; Dec 15; 165: 262-70.

38. Gupta V, Singh S, Gupta YK. Synthesis, characterization and antimicrobial activity of Co(II), Ni(II), Cu(II) and Zn(II) complexes N- O- S donor ligands. Asian J. Pharm. Ana. 2014; 4(4): 174-177.

39. Purohit MC, Kandwal A, Purohit R, A.R. Semwal, Shama Parveen, Arun K. Khajuria. Antimicrobial Activity of Synthesized Zinc Oxide Nanoparticles using Ajuga bracteosa Leaf Extract. Asian Journal of Pharmaceutical Analysis. 2021; 11(4): 275-0.

40. Hemalatha M, Arirudran B, Thenmozhi A, Mahadeva Rao US. Antimicrobial effect of separate extract of acetone, ethyl acetate, methanol and aqueous from leaf of milkweed (Calotropis gigantea L.). Asian J. Pharm. Res. 2011; 1(4): 102-107.

41. Killedar SG, More HN. Screening of antimicrobial potential and phytoconsituents for different extracts of Memecylon umbellatum Burm inflorescences. Asian J. Pharm. Res. 2011; Oct. - Dec. 1(4): 114-118.

42. Kumar SS, Melchias G, Ravikumar P, Chandrasekar R, Kumaravel P. Bioinspired synthesis of silver nanoparticles using Euphorbia hirta leaf extracts and their antibacterial activity. Asian J. Pharm. Res. 2014; 4(1): 39-43.

43. Shah SS, Gupta A, Karne S, Shinde B. Immunological evaluation of Artocarpus heterophyllus for determining its antimicrobial and anti-inflammatory activity. Asian J. Pharm. Res. 2017; 7(2): 106-110.

44. Mohite SA, Shah RR, Patel NR. Antimicrobial activity of leaves extracts of Jatropha curcas. Asian J. Pharm. Res. 2018; 8(1): 17-20.

45. Tabassum S, Ahmed W, Sharma PK, Pathan IK, Bhatia M, Khan M. In vivo and In vitro model for evaluation of antimicrobial activity: A Review. Asian Journal of Pharmaceutical Research. 2023; 13(3): 169-4.

46. Karthick K, Kumaravel P, Hemalatha P, Thamaraiselvi L. Mechanistic Aspects: Biosynthesis of Silver Nanoparticles from Proteus mirabilis and its antimicrobial study. Asian J. Res. Pharm. Sci. 2013; 3(3):-136.

47. Chandrawanshi NK, Tandia DK and Jadhav SK. Determination of antidiabetic property of organic and nonorganic solvent extracts of Schizophyllum commune. New Bio World A Journal of Alumni Association of Biotechnology. 2019; 1(1): 5-8.

48. Chandrawanshi NK, Tandia DK. Determination of antioxidant and antidiabetic potency of polar solvent extracts of Laetiporus sulphureus (Bull.) Murrill. Research Journal of Biotechnology. 2024; 19(7): 69-77.

Received on 22.01.2024 Revised on 17.05.2024

Accepted on 15.08.2024 Published on 24.12.2024

Available online from December 27, 2024

Research J. Pharmacy and Technology. 2024;17(12):5689-5696.

DOI: 10.52711/0974-360X.2024.00866

S. No	METHOD	*D. confragosa* (CH)	*D. confragosa* (FH)	*S. commune* (SCH)	*L. sulphurous* (TSH)
1	Reducing sugar (Fehling's test)	++	++	+	+++
2	Terpenoids (Salkowski test)	++	+++	+	+++
3	Flavonoids	++	+++	+	+++
4	Saponins	++	++	++	+++
5	Tannins	++	++	++	+++
6	Cardiac glycosides (Keller-Killiani test)	+	++	+	+++
7	Anthraquinones	+	+	++	+
8	Phlobatannins	+	+	+	++
9	Carotenoids	+	+	-	++
10	Phenols (Ferric chloride test)	+	++	++	+++

S.No	Mushroom samples	Habitat/ Collection site	Extraction solvent	Yield of Extract (% dry mushroom) 'g'
1	D. confragosa (C )	Parasitic growth on deciduous trees (Charama forest)	Water	50.00
			Methanol	2.7
			Ethanol	3.65
2	D. confragosa (F)	Parasitic growth on deciduous trees (Thakurtola forest)	Water	4.5
			Methanol	0.595
			Ethanol	1.505
3	S. commune (SC)	Parasitic growth on the trunk of Amla tree (Charama forest)	Water	19.6
			Methanol	14.00
			Ethanol	14.00
4	L.sulphureus (TS)	Parasitic growth on deciduous trees(Charama forest)	Water	9.8
			Methanol	11.1
			Ethanol	8.3