{"id":57784,"date":"2024-06-25T10:18:09","date_gmt":"2024-06-25T10:18:09","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=57784"},"modified":"2024-07-03T17:58:00","modified_gmt":"2024-07-03T17:58:00","slug":"evaluation-of-the-antioxidant-and-antimicrobial-activity-of-the-nutritionally-rich-plant-dioscorea-alata-l","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol17no2\/evaluation-of-the-antioxidant-and-antimicrobial-activity-of-the-nutritionally-rich-plant-dioscorea-alata-l\/","title":{"rendered":"Evaluation of the antioxidant and antimicrobial activity of the nutritionally rich plant, Dioscorea alata L."},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

Tubers\nare the storage organ of plants that store edible starch material as well as\nother nutrients and play a significant role in the contribution of dietary energy. Dioscorea<\/em>, <\/em>commonly\nknown as yam, is a vital tuber-yielding crop that produces about 10% of the\ntotal global production of roots and tubers.1<\/sup>Genus Dioscorea of the family\nDioscorea constitutes about 600 species throughout the world.2<\/sup> There are about 12 species found\nin Odisha, among which Dioscorea alata is cultivated and the other eleven are\nwild species.2<\/sup> Dioscorea alata <\/em>produces two types of\ntuber. One type produces under\nthe ground, known as an underground tuber while another type produces above the\nground on the axils of the stem, known as an aerial tuber. These are found in Tropical areas, North America,\nAfrica, Nepal, Indonesia, India, Japan, China, Mexico, Australia, South pacific islands, South\nAmerica, West Africa, and East Africa.2-4<\/sup> In India, D. alata<\/em> is largely\nfound the in the Eastern Ghats, Northeastern Himalayas and Western Ghats.2<\/sup> In Odisha, it is distributed all over the\nstate. It is rich in Similipal Biosphere Reserve, Phulbani district, Koraput, and\nMalkangiri districts of Odisha state.5<\/sup> D. alata<\/em> is the\nthird most major tuberous crop after cassava and sweet potato.4<\/sup> D.\nalata<\/em> L. is documented as Greater yam, Water yam, purple yam and Winged yam.3,6 <\/sup>In\ndifferent local languages identified as Kath also, Banra, Bandrara, Maati Aalu, Desia Aalu, Mate\nalu, Raja ala, Bebaru.2,3,6-10<\/sup> D. alata<\/em> is an annual and\nperennial climber plant of about 20-30 feet in height.3<\/sup> Its purple\ncolour stem has long petioles, bright green colour leaves and yellow-white\ncolour flowers.3<\/sup> The tuber of D. alata<\/em> is white in colour and\nwatery in texture.3<\/sup> The tubers of D. alata<\/em> are sources of many essential nutrients such as\ncarbohydrates, protein, vitamins and other nutrients.<\/strong>1,4,10<\/sup>This nutritional richer is\nused as a chief ingredient\nof traditional Odia food dalma2<\/sup>. Besides nutrient components,Dioscorea alata<\/em> also contains secondary\nmetabolites such as phenolic\nacid, flavonoids, coumarins, quinines, alkaloids, amines, terpenoids,\nphytosterols, tannin, diosgenin, and saponins.2, 3, 8-11<\/sup> D. alata<\/em> has been reported to exhibit\nantifungal, antidiabetic, antibacterial\nand antioxidant activities.2, 9,12-15<\/sup> Further D. alata<\/em> helps to cure piles, and stomach worm and demonstrate\nanti-diarrhoea, anti-inflammatory, antihypertensive, hypolipidemic and hypocholesteric\nactivity3<\/sup>. Juice\nof D. alata<\/em> is used as a cooling agent during summer.2<\/sup>\nTuber pastes of D. alata<\/em>\nare applied on cancerous wounds, leprosy, gonorrhoea and skin disease.8<\/sup>\nIn this present study nutritional values, DPPH scavenging potential and\nantimicrobial activity of both underground and aerial tuber of D. alata <\/em>were investigated.<\/em><\/strong><\/p>\n\n\n\n

Materials and Methods<\/strong><\/p>\n\n\n\n

Collection and\npreparation of sample<\/strong><\/p>\n\n\n\n

Freshly harvested elongated spherical\nshape, D. alata<\/em> tuber of good quality purchased\nfrom the local market of Burla, Odisha, India, in November 2018. The tubers\nwere thoroughly washed three to four times to remove adhering soil. Cleaned\ntubers were peeled and sliced about 1-2mm in thickness. Thinly sliced tubers\nwere oven dried at 80\u00b0<\/sup>C until a steady weight\nwas obtained. Then pulverized by a food processor (Usha FP 3811 Food Processor) and screened\nthrough a 1mm sieve to get the powder form of the sample. Tuber powder was kept\nin an airtight glass bottle for further analysis.<\/p>\n\n\n\n

Methods<\/strong><\/p>\n\n\n\n

Determination\nof moisture<\/strong><\/p>\n\n\n\n

The\nA.O.A.C method was used to determine the moisture content (1970).16<\/sup>\nThe samples were weighed carefully and dried at 80\u00b0C until they attained a\nconsistent weight. The moisture content was calculated using the following\nrelationship after the estimation was done in triplicate, and the mean values\nof both were recorded.<\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

Ash<\/strong><\/p>\n\n\n\n

The\nash content was determined by heating the food sample in a muffle furnace at 5500<\/sup>C.17<\/sup><\/p>\n\n\n\n

Carbohydrate<\/strong><\/p>\n\n\n\n

 The Anthrone reagent method was used to estimate carbohydrates. 100mg\nof material was hydrolyzed in 5 ml of 2.5N HCL for three hours. Then sodium carbonate\nwas added to this sample solution until the formation of effervescences ceased for neutralization\nand the volume was made up to 100ml before centrifugation. Anthrone reagent of 4ml was added to the\n0.5ml of supernatant. Afterwards, the reaction mixture was heated for eight\nminutes in a hot water bath and the intensity of the developed green colour was\nmeasured spectrophotometrically\nat 630nm.18<\/sup><\/p>\n\n\n\n

Starch<\/strong><\/p>\n\n\n\n

100mg\nof the sample was washed with hot 80% ethanol till the washing didn’t give in green\ncolour with Anthrone reagent to eliminate sugar from the sample. The residue\nwas dried over a water bath before being mixed with 5 mL of distilled water and\n52 % perchloric acid. Anthrone reagent of 4ml was added to 0.2 ml of supernatant and the\nreaction mixture was heated for eight minutes in a boiling water bath. After\nthat, the intensity of colour was measured at 630nm. The sample’s glucose\nconcentration was then multiplied by 0.9 to get the starch content.18<\/sup> <\/p>\n\n\n\n

Protein estimation<\/strong><\/p>\n\n\n\n

 Protein content was estimated by the Lowery et\nal., 1951 method. 19<\/sup>\nReagent C was a mixture of 50ml of reagent A (2 % sodium carbonate in 0.1N\nsodium hydroxide) and 1ml of reagent B (0.5 % copper sulphate in 1 % potassium\nsodium tartrate).  Reagent D contained 1\nmL Folin-Ciocalteau reagent and 1 mL distilled water. 1gm of dry powder sample\nwas homogenized with 10ml cold phosphate buffer (Ph 7.5, 0.1M). After\ncentrifugation, 5 ml of reagent C was added to 0.2 ml of sample extract,\nbrought up to 1 ml with water, and left for 10 minutes. Then 0.5ml of reagent D\nwas added to the reaction mixture and incubated for 30 minutes at room\ntemperature. At 660nm, the developed blue colour was measured.<\/p>\n\n\n\n

Fat <\/strong><\/p>\n\n\n\n

A powder sample of 5gm was transferred to a thimble plugged with a wad of fat-free cotton and dropped into the bottom of the extraction tube. The bottom of the extraction tube was connected to the Soxhlet flask and the top to the condenser. Before joining the flask weight of the empty flask was taken. In the extraction flask, 200ml of petroleum ether (Boiling point 400<\/sup>-500<\/sup>C) was poured. The extractions were continued for up to 16 hours. At the end of the extraction period, all the petroleum ether was evaporated and dried at 100\u00b0<\/sup>C for 1 hour, and it was cooled in desiccators and weighed.18<\/sup><\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

Ascorbic acid<\/strong><\/p>\n\n\n\n

The ascorbic acid solution was prepared\nby dissolving 5mg of ascorbic acid in 50 ml of4 % oxalic acid. The ascorbic acid solution was again mixed with\n10 mL of 4 % oxalic acid. The prepared solution was titrated against 2,\n6-dichlorophenolindophenol dye in sodium bicarbonate until the pink colour\ndeveloped, which lasted a few minutes (V1). A sample of 5gm was extracted in 4 %\noxalic acid and diluted to a volume of 100 mL before centrifugation. The\nsupernatant of 5 ml was mixed with 10 mL of 4 % oxalic acid and titrated\nagainst the dye (V2 ml). 18<\/sup> <\/p>\n\n\n\n

Calculation<\/p>\n\n\n\n

Amount\nof ascorbic acid (mg\/100g of sample)<\/p>\n\n\n\n

=0.5mg\/V1\nml \u00d7V2 ml\/5ml\u00d7100ml\/ weight.\nof sample \u00d7100<\/p>\n\n\n\n

Sodium and potassium\nestimation<\/strong><\/p>\n\n\n\n

Flame\nphotometry was used to determine sodium and potassium levels. KCL and NaCl were\nused to make standard solutions at different levels (0,5, and 10 ppm) of K and\nNa. A sample obtained by dry ashing was used to determine the total K and Na.\nFirst, the instrument was calibrated using a standard solution, and a standard\ncurve was created. The digest was diluted to the appropriate concentration\nrange, resulting in a final concentration of 0 to 5 mg\/kg. The samples were\nthen examined at 768 nm in a flame photometer. 17<\/sup><\/p>\n\n\n\n

Phosphorus <\/strong><\/p>\n\n\n\n

The\nphosphomolybdate technique was used to calculate phosphorus. 1ml of molybdate\nreagent (6.0g of ammonium molybdate was dissolved in 40ml of water, then 50ml\nof 10N H2SO4 was added, bringing the total amount to 100 ml) was added to 1ml of ash solution\nprepared by dry ashing. After that, 0.4ml of amino naphthol sulphonic acid solution was added, increasing\nthe volume to 10ml. A blank was made the same way but using water instead of\nthe sample. It was allowed to stand for 10 minutes before being tested at 650nm.17<\/sup><\/p>\n\n\n\n

Iron<\/strong><\/p>\n\n\n\n

The amount of iron in\nthe sample was assessed by oxidizing it with potassium persulphate and then\ntreating it with potassium thiocyanate to produce red ferric thiocyanate, which\nwas quantified calorimetrically at 480nm.17<\/sup><\/p>\n\n\n\n

Phytochemicals estimation<\/strong><\/p>\n\n\n\n

Total phenolic content<\/strong><\/p>\n\n\n\n

Total phenolic content was determined\nby a slightly modified method of Oueslati et al., 2012.20<\/sup> Distilled water (0.5ml) and\nFolin-Ciocalteu reagent (0.2ml) were added to the 0.5ml sample extract. The\nmixture was mixed properly and left for 6min. After that 0.2ml of 7% Na2<\/sub>CO3   <\/sub>was\nadded to it. The final volume was made up to 3ml and incubated for 90 minutes\nin a dark place. Absorbance was measured at 760nm. The total phenolic content\nwas calculated as mg of gallic\nacid equivalents per 100g of dry mass through a calibration curve with gallic acid.<\/p>\n\n\n\n

Flavonoid estimation<\/strong><\/p>\n\n\n\n

The\nflavonoid content of the sample was determined using the method of Kamtekar et al., 2014\nwith slight changes.21<\/sup>\nIn a test tube, 1 mL of aliquots and 1 ml of quercetin solution received 4 mL of distilled\nwater and 0.3 mL of 5% sodium nitrite solution. After 5mintues 0.3 mL of 10%\naluminium chloride was added. Further 2 mL of 1M sodium hydroxide was added at 6\nminutes.  At 510 nm, the intensity of the\nyellowish-orange colour was measured. The flavonoid content was calculated as\nmg of quercetin\nequivalents per 100g of dry mass.<\/p>\n\n\n\n

Tannin\nestimation<\/strong><\/p>\n\n\n\n

Tannin\nwas calculated using Schanderl’s\ntechniques (1970).22<\/sup>\nThe powder sample (0.5g)\nwas boiled in 75ml water for 30 minutes and centrifuged at 2,000rpm for 20\nminutes, the supernatant was collected, and a final volume of 100ml was\nobtained. From this, 0.1ml extract was taken with 7.5ml water, 0.5ml\nFolin-Denis reagent, and 1ml sodium carbonate solution was added, and a final\nvolume of 10ml was obtained. After 30 minutes, the absorbance was measured at\n700nm. Tannic acid equivalents are used to express the sample’s tannin content.<\/p>\n\n\n\n

Diosgenin estimation<\/strong><\/p>\n\n\n\n

 One gram of material was mixed with 30 ml of methanol\nand agitated overnight. The supernatant was obtained after centrifuging the\nextract for 18 minutes at 3500 rpm. The supernatants were obtained after two\nfurther extractions. The ultimate capacity was set at 100 milliliters. A tube\nwas filled with 0.1ml of the methanol extract, evaporating under decreased\npressure. The residue was diluted in 2 mL ethyl acetate, 1 mL each of reagent A\n(0.5 mL p-anisaldehyde in 99.5 mL ethyl acetate) and B (50 mL concentrated H2SO4\n+ 50 mL ethyl acetate) was added and thoroughly mixed. The test tube was\ninserted into a water bath and kept at 600 degrees for 10 minutes to generate a\ncolour. After that, it was allowed to cool in a 250\u00b0F water bath for 10\nminutes. A spectrophotometer was used to measure the absorbance at 430nm. 2ml\nethyl acetate was put in a tube and tested in the same way as the reagent blank.23, 24<\/sup><\/p>\n\n\n\n

DPPH\nradical scavenging assay<\/strong><\/p>\n\n\n\n

DPPH free radical scavenging activity of the sample extract was evaluated using the method of Sakthidevi and Mohan, 2013 with slight modification.25<\/sup> DPPH (1ml of 0.1mm) solution in methanol was added to 3ml of sample extract and ascorbic acid solution in methanol at different concentrations (100, 200, 400, 800 \u00b5g\/ml). Ascorbic acid was taken as a reference. Absorbance was measured at 517nm. A lower absorbance value refers to higher DPPH scavenging activity. The following formula was used to calculate DPPH scavenging activity.<\/p>\n\n\n\n

DPPH scavenging activity (% of inhibition) = (A0-A1)\/A0 \u00d7100<\/p>\n\n\n\n

Where A0= Absorbance of the control, A1= Absorbance of sample and reference<\/p>\n\n\n\n

Antimicrobial\nactivity<\/strong><\/p>\n\n\n\n

Following the conventional procedure\noutlined, the powder sample was extracted sequentially using six organic\nsolvents (methanol, chloroform, ethyl acetate, acetone, water, and petroleum\nether).26<\/sup> The\nextracts were centrifuged, filtered, and concentrated further in a vacuum rotary evaporator\n(model with evaporation\ncondition). The antibacterial activity of the sticky layers was tested\nby dissolving them in 100 mg\/ml DMSO. The antibacterial activities of different\nsolvents and aqueous extracts of D. alata<\/em>\nwere determined using the agar well diffusion method, slightly modified from\nNavarro et al. (1966). Each Petri plate received about 25 mL of nutritional\nagar. Pathogenic multidrug-resistant bacteria (MDR) cultures such as A. baumannii<\/em>, E. faecalis<\/em>, K. pneumonia<\/em>,\nand P. mirabilis<\/em> were added to the\nagar once it had solidified. The antibacterial activity was measured in\ntriplicate and expressed as the mean of inhibition. The sample’s minimal\ninhibitory concentration (MIC) and minimal bactericidal concentration (MBC)\nwere determined a using previously published method.27<\/sup><\/p>\n\n\n\n

Toxicity evaluation<\/strong><\/p>\n\n\n\n

Methanolic\nextract of D. alata<\/em> was evaluated for\nits acute and sub-acute toxicity following the OECD guidelines 423 and 407,\nrespectively. The animal\nexperiment was conducted at the School of Pharmacy, Siksha \u2018O\u2019 Aunsandhan\nUniversity, and the protocol used was approved by the Animal Ethics Committee\n(Protocol IAEC\/SPS\/SOA\/17\/2018). Twenty-four male albino Wister rats\n(180-230g), aged 7-8 weeks, were divided into six rats, one control group, and\nthree treated groups. Animals were kept in a temperature-controlled environment\n(23 \u00b1 20 o<\/sup>C) with a 12-hour light-dark cycle. The control group\nreceived water only, and each treated group received a single oral dose of\nextract. Methanolic extract of D. alata<\/em>\nwas given in 2000 mg\/kg, 4000 mg\/kg and 8000 mg\/kg body weight. After\nadministering the extract, the animals were sectioned for changes in their\ngeneral behaviour, physiological activities and survival for 72 hours in acute\ntoxicity evaluation. In subacute toxicity analysis, the t, related group received\nextracts for 45 days and\nthe animals were anaesthetised with formalin. The animals were sacrificed to\ncollect their blood and organs (liver, kidney) for biochemical and histological\nanalysis. On day 46, the control and treated groups o were given an overdose (0.2 ml) of 3.5% formaldehyde. Then blood was\ntaken from the heart for analysis. The biochemical parameters analysed\nfrom serum were glucose (G), total cholesterol (TC), triglycerides (TG),\naspartate aminotransferase (AST), alanine aminotransferase (ALT), urea (Ur),\ncreatinine (Cr) and total protein. The\norgans, liver, and kidney were removed and embedded in paraffin, sectioned and\nstained with hematoxylin and eosin. The tissues were observed under the\nmicroscope for histopathological toxicity evaluation.<\/p>\n\n\n\n

Statistical\nAnalysis<\/strong><\/p>\n\n\n\n

The results obtained were subjected to\nstatistical analysis as mean and standard deviation.28<\/sup> The mean values and standard\ndeviations were calculated from the data obtained from three different\nexperiments. The statistical difference at p < 0.05 was considered to be significant.<\/strong> Analysis\nof variance (ANOVA) was subjected within the animal groups for obtained data of\neach biomedical parameter in toxicity analysis.<\/p>\n\n\n\n

Result <\/strong><\/p>\n\n\n\n

Nutritional components<\/strong><\/p>\n\n\n\n

Nutritional components analysis included moisture, ash, carbohydrate, starch, fat, protein, ascorbic acid and minerals (Figures 1 & 2; Table 1). The underground tuber\u2019s moisture content was high (66.22%) than the areal tuber (58.94%). Areal tuber was significantly higher at p<0.05 ash (4.64%) than underground tuber (2.68%). The Carbohydrate Content of the areal tuber was estimated to be reasonably low (42.08%) compared to the underground tuber (58.96%). The starch content of the underground tuber was higher (5.61%) than the areal tuber (3.26%). The Underground and areal tuber contained the nearly same amount of fat (0.30%-0.33%). The protein content was higher (1.39%) in the aerial tuber compared to the underground tuber (0.78%). The amino acid content of the underground tuber was estimated to be quite high (2.49%) compared to the areal tuber (0.8%). The ascorbic acid content of the areal tuber on a dry weight basis was significantly low at p<0.05 (45mg\/100gm) compared to the underground tuber (87.34mg\/100gm). Analysis of minerals on a dry weight basis included sodium, potassium, iron and phosphorus. Sodium, potassium, iron and phosphorus contents of underground tuber were found to be 51.38mg\/100gm, 206.33mg\/100gm, 129.5mg\/100gm and 20.21mg\/100gm respectively while areal tuber contained 39.08mg\/100gm, 195.23mg\/100gm, 118.4mg\/100gm and 22.02mg\/100gm of sodium, potassium, iron and phosphorus respectively. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 1: Nutrients contents of the underground and areal tuber of D. alata<\/em><\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Table 1: Nutritional compositions of the underground and aerial tuber of D. alata<\/em><\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Parameter<\/strong><\/p>\n<\/td>\n

\n

DA(Underground)<\/strong><\/p>\n<\/td>\n

\n

DA(Aerial)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Moisture<\/strong><\/p>\n<\/td>\n

\n

59.73\u00b10.88<\/p>\n<\/td>\n

\n

68.51\u00b11.5<\/p>\n<\/td>\n<\/tr>\n

\n

Ash<\/strong><\/p>\n<\/td>\n

\n

2.68\u00b11.53<\/p>\n<\/td>\n

\n

4.64\u00b12.4<\/p>\n<\/td>\n<\/tr>\n

\n

Carbohydrate<\/strong><\/p>\n<\/td>\n

\n

58.96\u00b10.02<\/p>\n<\/td>\n

\n

42.08\u00b10.05<\/p>\n<\/td>\n<\/tr>\n

\n

Starch<\/strong><\/p>\n<\/td>\n

\n

3.26\u00b10.01<\/p>\n<\/td>\n

\n

5.61\u00b10.007<\/p>\n<\/td>\n<\/tr>\n

\n

Reducing sugar<\/strong><\/p>\n<\/td>\n

\n

0.014\u00b10.005<\/p>\n<\/td>\n

\n

0.029\u00b10.007<\/p>\n<\/td>\n<\/tr>\n

\n

Fat<\/strong><\/p>\n<\/td>\n

\n

0.30\u00b10.7<\/p>\n<\/td>\n

\n

0.33\u00b10.28<\/p>\n<\/td>\n<\/tr>\n

\n

Protein<\/strong><\/p>\n<\/td>\n

\n

0.78\u00b10.01<\/p>\n<\/td>\n

\n

1.39\u00b10.22<\/p>\n<\/td>\n<\/tr>\n

\n

Free amino acid<\/strong><\/p>\n<\/td>\n

\n

2.49\u00b10.48<\/p>\n<\/td>\n

\n

0.8\u00b10.9<\/p>\n<\/td>\n<\/tr>\n

\n

Ascorbic acid (mg\/100gm)<\/strong><\/p>\n<\/td>\n

\n

87.43\u00b11.2<\/p>\n<\/td>\n

\n

45\u00b10.05<\/p>\n<\/td>\n<\/tr>\n

\n

Sodium(mg\/100gm)<\/strong><\/p>\n<\/td>\n

\n

51.38\u00b12.56<\/p>\n<\/td>\n

\n

39.08\u00b11.67<\/p>\n<\/td>\n<\/tr>\n

\n

Potassium(mg\/100gm)<\/strong><\/p>\n<\/td>\n

\n

206.33\u00b12.51<\/p>\n<\/td>\n

\n

195.23\u00b12.02<\/p>\n<\/td>\n<\/tr>\n

\n

Iron(mg\/100gm)<\/strong><\/p>\n<\/td>\n

\n

129.5\u00b13.11<\/p>\n<\/td>\n

\n

118.4\u00b12.13<\/p>\n<\/td>\n<\/tr>\n

\n

Phosphorus(mg\/100gm)<\/strong><\/p>\n<\/td>\n

\n

20.21\u00b12.12<\/p>\n<\/td>\n

\n

22.02\u00b11.95<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Note: Each value is the average of three analyses \u00b1 standard deviation.<\/p>\n\n\n\n
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Figure 2: Vitamins and Minerals contents of the underground and areal tuber  of D. alata <\/em>on a dry weight basis<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Bioactive components:<\/strong><\/p>\n\n\n\n

The\nFlavonoid content of underground and areal tuber of D. alata<\/em> was estimated to be 390mg\/100gm and 273mg\/100gm, respectively\non a dry weight basis\n(Table 2; Figure 3). The total phenolic content of the underground tuber was\nfound to be significantly\nhigh at p<0.05 (248.30mg\/100gm) compared to the areal tuber (130mg\/100gm)\n(Figure 3). The Diosgenin content of the underground tuber was found to be relatively\nlow (50.87mg\/100gm) compared to the areal tuber (89.67mg\/100gm) (Figure 3).\nTannin content was 451.23mg\/100gm and 721.06mg\/100gm for an underground and\naerial tuber of D. alata<\/em>, respectively (Figure 3).<\/p>\n\n\n\n

Table 2: Bioactive components of D. alata<\/em> tubers<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\n

Parameters<\/strong><\/p>\n<\/td>\n

\n

Underground<\/strong><\/p>\n<\/td>\n

\n

Aerial<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Flavonoid(mg\/100gm)<\/strong><\/p>\n<\/td>\n

\n

390 \u00b12.1<\/p>\n<\/td>\n

\n

273\u00b10.8<\/p>\n<\/td>\n<\/tr>\n

\n

Phenol(mg\/100gm)<\/strong><\/p>\n<\/td>\n

\n

248.30\u00b11.9<\/p>\n<\/td>\n

\n

130\u00b11.1<\/p>\n<\/td>\n<\/tr>\n

\n

Tannin(mg\/100gm)<\/strong><\/p>\n<\/td>\n

\n

451.23\u00b11.7<\/p>\n<\/td>\n

\n

721.06\u00b11.2<\/p>\n<\/td>\n<\/tr>\n

\n

Diosgenin(mg\/100gm)<\/strong><\/p>\n<\/td>\n

\n

50.87\u00b12.2<\/p>\n<\/td>\n

\n

89.67\u00b11.3<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Each value is the average of three analyses \u00b1 standard deviation.<\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 3: Bioactive components of the underground and areal tuber of D. alata <\/em>in dry weight basis.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Determination\nof antioxidant activity by DPPH scavenging activity<\/strong><\/p>\n\n\n\n

The IC50 value is defined as the amount of sample necessary to decrease the absorbance of DPPH by 50 %. The IC50 value of methanolic extract of the underground and areal tuber of D. alata<\/em> was 121.81 \u00b5g\/ml and 324.28 \u00b5g\/ml, respectively. The underground tuber possessed a lower value of IC50 than the areal tuber, which indicated the underground tuber exhibited a higher potential for DPPH scavenging activity than the areal tuber. Figure 4 represents the DPPH scavenging activity of the underground and areal tuber of D. alata<\/em>.<\/p>\n\n\n\n

Table 3: IC50 values of D. alata <\/em>tubers<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

Concentration<\/strong><\/p>\n

(\u00b5g\/ml)<\/strong><\/p>\n<\/td>\n

\n

Underground<\/strong><\/p>\n<\/td>\n

\n

Aerial<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

100<\/strong><\/p>\n<\/td>\n

\n

33.8<\/p>\n<\/td>\n

\n

29.6<\/p>\n<\/td>\n<\/tr>\n

\n

200<\/strong><\/p>\n<\/td>\n

\n

58.2<\/p>\n<\/td>\n

\n

39.8<\/p>\n<\/td>\n<\/tr>\n

\n

400<\/strong><\/p>\n<\/td>\n

\n

95.2<\/p>\n<\/td>\n

\n

58.9<\/p>\n<\/td>\n<\/tr>\n

\n

600<\/strong><\/p>\n<\/td>\n

\n

95.7<\/p>\n<\/td>\n

\n

75.5<\/p>\n<\/td>\n<\/tr>\n

\n

800<\/strong><\/p>\n<\/td>\n

\n

97.1<\/p>\n<\/td>\n

\n

86.5<\/p>\n<\/td>\n<\/tr>\n

\n

IC50<\/strong><\/p>\n<\/td>\n

\n

121.81<\/p>\n<\/td>\n

\n

324.28<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 4: DPPH scavenging activity of underground and areal tuber of D. alata<\/em><\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Antimicrobial property<\/strong><\/strong><\/p>\n\n\n\n

The antibacterial activity of six\nsolvent extracts was tested using the agar well diffusion method on independent\nlawn cultures of seven bacterial strains (2 GPs and 5 GNs). Acetone extracts\nhad the most significant inhibitory zones against MRSA (29 mm) and P. mirabilis\n(29 mm). Similarly, the inhibitory zone against VRE was the largest in\nmethanolic extract (29 mm). The petroleum-ether extract and aqueous tuber\nextract demonstrated deficient antibacterial activity than the other four\nsolvent extracts. Antibacterial activity was evaluated on all other solvent\nextracts (Table 4).<\/p>\n\n\n\n

The maximal\nantibacterial activity was evaluated by determining acetone and methanolic\nextracts’ MIC and MBC values. A MIC value of 3.0 mg\/mL of acetone extract was\nregistered against A. baumannii<\/em>, E. fecalis<\/em>, S. aureus,<\/em> S. pyogenes<\/em>, and\nP. mirabilis<\/em>; 6.0 mg\/mL as MIC\nagainst P. aeruginosa<\/em> and K. pneumonia<\/em> was<\/em> recorded; Similarly, the MIC value of 6.0 mg\/mL of methanolic\nextract was registered against A.\nbaumannii<\/em>, K.\npneumoniae<\/em> E. fecalis<\/em>, S. pyogenes<\/em>; 3.0 mg\/mL against S. aureus, P. aeruginosa<\/em> and P. mirabilis<\/em> (Table 5). Further, the MBC\nvalues of these two active extracts were determined. An MBC value of 15.0 mg\/mL\nof acetone extract wasregistered\nagainst A. baumannii and E. fecalis,<\/em> and\na discount of 25 mg\/mL against S. aureus,<\/em> P. mirabilis <\/em>and K. pneumonia<\/em> was recorded, and 50 mg\/mL against P. aeruginosa <\/em>was recorded.<\/em> Similarly, an MBC value of 15.0 mg\/mL\nof methanolic extract was registered against P. mirabilis<\/em>; a 25 mg\/mL discount against A. baumannii<\/em>, E. faecalis<\/em>, S. pyogenes<\/em>, and K.\npneumonia<\/em> was reported; and a value of 50 mg\/mL against S. aureus <\/em>was recorded.<\/p>\n\n\n\n

Table 4: Six hot solvents D. alata<\/em> antimicrobial assays using the agar well diffusion method against MDR bacterial strains (zone of inhibition in mm).<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Clinical<\/strong><\/p>\n

Strain<\/strong><\/p>\n<\/td>\n

\n

Petroleum ether<\/strong><\/p>\n<\/td>\n

\n

Chloro-<\/strong><\/p>\n

form<\/strong><\/p>\n<\/td>\n

\n

Ethyl acetate<\/strong><\/p>\n<\/td>\n

\n

Acetone<\/strong><\/p>\n<\/td>\n

\n

Methanol<\/strong><\/p>\n<\/td>\n

\n

Water<\/strong><\/p>\n<\/td>\n

\n

Linezolid\/imipenem<\/strong><\/p>\n

(30\/10 mg\/mL<\/strong>)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

A. baumannii<\/em><\/strong><\/p>\n<\/td>\n

\n

10\u00b1 1.33<\/p>\n<\/td>\n

\n

21\u00b11.37<\/p>\n<\/td>\n

\n

13\u00b11.21<\/p>\n<\/td>\n

\n

33\u00b11.13<\/p>\n<\/td>\n

\n

27\u00b11.21<\/p>\n<\/td>\n

\n

15\u00b11.31<\/p>\n<\/td>\n

\n

29\u00b10.89<\/p>\n<\/td>\n<\/tr>\n

\n

E. faecalis<\/em><\/strong><\/p>\n<\/td>\n

\n

15\u00b12.37<\/p>\n<\/td>\n

\n

22\u00b11.33<\/p>\n<\/td>\n

\n

23\u00b11.12<\/p>\n<\/td>\n

\n

28\u00b11.61<\/p>\n<\/td>\n

\n

26\u00b10.93<\/p>\n<\/td>\n

\n

22\u00b11.37<\/p>\n<\/td>\n

\n

29\u00b10.37<\/p>\n<\/td>\n<\/tr>\n

\n

K. pneumoniae<\/em><\/strong><\/p>\n<\/td>\n

\n

17\u00b11.33<\/p>\n<\/td>\n

\n

15\u00b11.24<\/p>\n<\/td>\n

\n

14\u00b11.59<\/p>\n<\/td>\n

\n

26\u00b10.53<\/p>\n<\/td>\n

\n

29\u00b10.51<\/p>\n<\/td>\n

\n

13\u00b11.15<\/p>\n<\/td>\n

\n

33\u00b11.23<\/p>\n<\/td>\n<\/tr>\n

\n

P. mirabilis<\/em><\/strong><\/p>\n<\/td>\n

\n

08\u00b11.78<\/p>\n<\/td>\n

\n

12\u00b11.15<\/p>\n<\/td>\n

\n

12\u00b11.04<\/p>\n<\/td>\n

\n

29\u00b11.87<\/p>\n<\/td>\n

\n

19\u00b11.15<\/p>\n<\/td>\n

\n

13.5\u00b11.83<\/p>\n<\/td>\n

\n

31\u00b11.73<\/p>\n<\/td>\n<\/tr>\n

\n

P. aeruginosa<\/em><\/strong><\/p>\n<\/td>\n

\n

11\u00b11.53<\/p>\n<\/td>\n

\n

18\u00b10.57<\/p>\n<\/td>\n

\n

14\u00b11.27<\/p>\n<\/td>\n

\n

27\u00b11.39<\/p>\n<\/td>\n

\n

21\u00b11.27<\/p>\n<\/td>\n

\n

13\u00b10.67<\/p>\n<\/td>\n

\n

26\u00b11.21<\/p>\n<\/td>\n<\/tr>\n

\n

S. aureus <\/em><\/strong>(MRSA)<\/strong><\/p>\n<\/td>\n

\n

10\u00b11.01<\/p>\n<\/td>\n

\n

19\u00b11.93<\/p>\n<\/td>\n

\n

14\u00b11.53<\/p>\n<\/td>\n

\n

26\u00b11.73<\/p>\n<\/td>\n

\n

23\u00b11.33<\/p>\n<\/td>\n

\n

14\u00b11.19<\/p>\n<\/td>\n

\n

29\u00b10.73<\/p>\n<\/td>\n<\/tr>\n

\n

S. pyogenes<\/em><\/strong><\/p>\n<\/td>\n

\n

18\u00b10.53<\/p>\n<\/td>\n

\n

18\u00b10.89<\/p>\n<\/td>\n

\n

15\u00b11.21<\/p>\n<\/td>\n

\n

29\u00b11.91<\/p>\n<\/td>\n

\n

22\u00b11.97<\/p>\n<\/td>\n

\n

17\u00b10.79<\/p>\n<\/td>\n

\n

26\u00b11.51<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Each value is the average of three analyses \u00b1 standard deviation.<\/p>\n\n\n

Table 5: MIC and MBC of two bioactive fractions of D. alata <\/em>against MDR bacterial strains (mg\/ml).<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Strain<\/strong><\/p>\n<\/td>\n

\n

Acetone<\/strong><\/p>\n<\/td>\n

\n

Methanol<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

MIC<\/strong><\/p>\n<\/td>\n

\n

MBC<\/strong><\/p>\n<\/td>\n

\n

MIC<\/strong><\/p>\n<\/td>\n

\n

MBC<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

A. baumannii<\/em><\/strong><\/p>\n<\/td>\n

\n

3.0<\/p>\n<\/td>\n

\n

12<\/p>\n<\/td>\n

\n

6.0<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n<\/tr>\n

\n

E. faecalis<\/em> (VRE)<\/strong><\/p>\n<\/td>\n

\n

3.0<\/p>\n<\/td>\n

\n

15.0<\/p>\n<\/td>\n

\n

6.0<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n<\/tr>\n

\n

K. pneumoniae<\/em><\/strong><\/p>\n<\/td>\n

\n

6.0<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n

\n

6.0<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n<\/tr>\n

\n

P. mirabilis<\/em><\/strong><\/p>\n<\/td>\n

\n

3.0<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n

\n

1.5<\/p>\n<\/td>\n

\n

15.0<\/p>\n<\/td>\n<\/tr>\n

\n

P. aeruginosa<\/em><\/strong><\/p>\n<\/td>\n

\n

6.0<\/p>\n<\/td>\n

\n

50<\/p>\n<\/td>\n

\n

3.0<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n<\/tr>\n

\n

S. aureus <\/em>(MRSA)<\/strong><\/p>\n<\/td>\n

\n

3.0<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n

\n

6.0<\/p>\n<\/td>\n

\n

50<\/p>\n<\/td>\n<\/tr>\n

\n

S. pyogenes<\/em><\/strong><\/p>\n<\/td>\n

\n

3.0<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n

\n

6.0<\/p>\n<\/td>\n

\n

25<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

MIC: Minimal inhibitory concentration, MBC: Minimal bactericidal concentration<\/p>\n\n\n

Acute toxicity study <\/strong><\/p>\n\n\n\n

Oral\nadministration of the D. alata<\/em>\nmethanolic extract (2000 to 8000 mg\/kg body weight) neither caused any death\nnor produced significant changes in the spontaneous type, alertness, awareness,\ngood response, touch response, pain response, righting reflex, pinna reflex,\ngrip strength in the experimental rats, during 72 hours of the testing period.\nAll groups of animals showed neither any toxic effect nor any lethal effect.\nAdministration of doses up to 8000 mg\/kg body weight of D. alata methanolic<\/em> extract did not reveal any toxicity or\nmortality in rats during the entire observation period. Therefore, the LD50\n<\/sub>of D. alata<\/em> methanolic extract\nmay be greater than 8000 mg\/kg.<\/p>\n\n\n\n

Sub-acute toxicity<\/strong><\/p>\n\n\n\n

Blood\nbiochemical parameters and histopathology of the kidney\nand liver of the control and experimental\nanimals were observed to investigate any side effects on the animal. <\/p>\n\n\n\n

Biochemical\nparameters<\/strong><\/p>\n\n\n\n

The\nblood biochemical parameters between treated and untreated animals were analysed\nto examine whether the D. alata<\/em>\nmethanolic extract has any side effects on animals. The data were collated in\nTable 6 as well as Figure 5.  Briefly,\nthe parameters examined include glucose, urea, creatinine, protein,\ncholesterol, triglycerides, aspartate aminotransferase (AST) and alanine aminotransferase\n(ALT). After 45 days of daily doses of methanolic extract of D. alata<\/em> failed to reveal any\nsignificant difference (using a one-way ANOVA test at p\u22640.05) in various blood\nbiochemical parameters between treated and untreated groups, indicating no side\neffects to animals. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 5: Blood biochemical parameters between treated and untreated groups of animals<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Table 6: Blood biochemical parameters of the control and treated groups of animals with boiled methanolic extracts of D. alata<\/em> tuber. <\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Parameters<\/strong><\/p>\n<\/td>\n

\n

Group-I<\/strong><\/p>\n<\/td>\n

\n

Group-II<\/strong><\/p>\n<\/td>\n

\n

Group-III<\/strong><\/p>\n<\/td>\n

\n

Group-IV<\/strong><\/p>\n<\/td>\n

\n

Normal range<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Glucose(mg\/dl)<\/strong><\/p>\n<\/td>\n

\n

78.46\u00b10.46<\/p>\n<\/td>\n

\n

75.24\u00b10.12<\/p>\n<\/td>\n

\n

76.10\u00b10.05<\/p>\n<\/td>\n

\n

75.24\u00b10.08<\/p>\n<\/td>\n

\n

70-110<\/p>\n<\/td>\n<\/tr>\n

\n

Urea(mg\/dl)<\/strong><\/p>\n<\/td>\n

\n

35.60\u00b10.62<\/p>\n<\/td>\n

\n

32.37\u00b10.18<\/p>\n<\/td>\n

\n

31.84\u00b10.29<\/p>\n<\/td>\n

\n

34.21\u00b10.28<\/p>\n<\/td>\n

\n

15-45<\/p>\n<\/td>\n<\/tr>\n

\n

Creatinine(mg\/dl)<\/strong><\/p>\n<\/td>\n

\n

0.86\u00b10.05<\/p>\n<\/td>\n

\n

0.85\u00b10.03<\/p>\n<\/td>\n

\n

0.83\u00b10.01<\/p>\n<\/td>\n

\n

0.83\u00b10.007<\/p>\n<\/td>\n

\n

0.5-1.5<\/p>\n<\/td>\n<\/tr>\n

\n

Total protein(mg\/dl) <\/strong><\/p>\n<\/td>\n

\n

6.56\u00b10.11<\/p>\n<\/td>\n

\n

6.81\u00b10.099<\/p>\n<\/td>\n

\n

6.83\u00b10.05<\/p>\n<\/td>\n

\n

6.92\u00b10.10<\/p>\n<\/td>\n

\n

6.0-8.0<\/p>\n<\/td>\n<\/tr>\n

\n

Total cholesterol (mg\/dl)<\/strong><\/p>\n<\/td>\n

\n

165.1\u00b10.99<\/p>\n<\/td>\n

\n

167.5\u00b10.53<\/p>\n<\/td>\n

\n

158.4\u00b10.74<\/p>\n<\/td>\n

\n

161.2\u00b10.88<\/p>\n<\/td>\n

\n

140-250<\/p>\n<\/td>\n<\/tr>\n

\n

Tri glycerides(mg\/dl)<\/strong><\/p>\n<\/td>\n

\n

92.12\u00b10.83<\/p>\n<\/td>\n

\n

113.5\u00b11.77<\/p>\n<\/td>\n

\n

113.1\u00b10.83<\/p>\n<\/td>\n

\n

115\u00b11.06<\/p>\n<\/td>\n

\n

25-160<\/p>\n<\/td>\n<\/tr>\n

\n

(AST)(IU\/L)<\/strong><\/p>\n<\/td>\n

\n

32.37\u00b10.51<\/p>\n<\/td>\n

\n

37.24\u00b10.13<\/p>\n<\/td>\n

\n

39.08\u00b10.22<\/p>\n<\/td>\n

\n

42.05\u00b10.02<\/p>\n<\/td>\n

\n

Up to 46<\/p>\n<\/td>\n<\/tr>\n

\n

(ALT)(IU\/L)<\/strong><\/p>\n<\/td>\n

\n

27.46\u00b10.41<\/p>\n<\/td>\n

\n

26.58\u00b11.42<\/p>\n<\/td>\n

\n

31.53\u00b10.67<\/p>\n<\/td>\n

\n

34.05\u00b10.03<\/p>\n<\/td>\n

\n

Up to 40<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

The values are mean \u00b1 standard deviation. The\ndifferences in various blood biochemical parameters are statistically\ninsignificant using one away ANOVA test among control and treated groups of the\nanimal at p\u22640.05). Group-I (Control), Group-II (2000 mg\/kg\nbody weight), Group-III (4000 mg\/kg body weight), Group-IV (8000 mg\/Kg\nbody weight), AST–<\/strong>Asparateamino transferase,\nALT- Alanine aminotransferase.<\/p>\n\n\n\n

Histopathological\nstudies<\/strong><\/p>\n\n\n\n

The\neffect of the methanolic\nextract of D.\nalata<\/em> tuber on the histological changes of kidney and liver tissues after\n45 days of the treatment has been shown below. The therapy with daily doses of\n2000, 4000 and 8000 mg\/kg body weight for 45 days failed to reveal any significant\nchanges compared to the control group. Necrosis, infiltration, oedema and\nconjunction, which are signs of hepatotoxicity, were not observed in the liver\ncells of the experimental group.  The liver showed standard hepatic lobular\narchitecture. The kidneys revealed normal glomeruli, proximal and distal\ntubules, interstitium, and blood vessels. The\nhistopathological images of the kidney and liver of control and treated with\ndifferent doses are shown in Figure 6. <\/p>\n\n\n

<\/p>\n\n\n\n
\"\"<\/td>\nFigure 6: Panels represent H&E staining of paraffin-embedded five-micron-thick sections of the kidney and liver at magnifications 200x of control and treated animals with an increasing dose of D. alata <\/em>tuber methanolic extract. <\/strong>

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Table 7: Effect of methanolic extract of Dioscorea alata<\/em> on acute toxicity.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Behaviour <\/strong><\/p>\n

Type<\/strong><\/p>\n<\/td>\n

\n

                                           Treatments<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Control<\/strong><\/p>\n<\/td>\n

\n

2000 mg\/kg body weight<\/strong><\/p>\n<\/td>\n

\n

4000 mg\/kg body weight<\/strong><\/p>\n<\/td>\n

\n

8000 mg\/kg body weight<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Spontaneous type<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Alertness<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Awareness<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Sound response<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Touch response<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Pain response<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Righting reflex<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Pinna reflex<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Grip strength<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Food intake<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Water intake<\/strong><\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n

\n

N<\/p>\n<\/td>\n<\/tr>\n

\n

Mortality<\/strong><\/p>\n<\/td>\n

\n

Ab<\/p>\n<\/td>\n

\n

Ab<\/p>\n<\/td>\n

\n

Ab<\/p>\n<\/td>\n

\n

Ab<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Discussion<\/strong><\/p>\n\n\n\n

Moisture\ncontent represents the water present in the tuber. Water physically interacts\nwith protein, polysaccharides, and lipids and influences texture, appearance,\nand flavour. But high moisture content affects the keeping quality of tubers.29<\/sup> The\nmoisture content of D. alata<\/em> was\nsignificantly higher in the present study than the values recorded in the available\nliterature.4,30,31\n<\/sup>However, another study reported a meagre amount of moisture content for D. alata.<\/em>32<\/sup>This variation in moisture\ncontent between the present study and other reports might be due to the level\nof maturity of tubers, geographical regions of cultivation, and methods used\nfor estimation. Ash is the inorganic food residue remaining after\nheating destroys organic matter. Inorganic components within a food represent\nminerals such as Na, K, Ca, Mg, Mn, P, Fe, Zn, Cu etc. Similar ash content has\nbeen reported in a previous study.31<\/sup> At the same time, the ash value reported in other studies\ndiffers remarkably from the present study. 4,33<\/sup> The ash content of tubers varied due to\nsoil, harvesting time and moisture content.31<\/sup> Carbohydrates are the primary energy source\nin the body. Fauziah et al., 2020 reported a low amount (17.10-29.37%) of\ncarbohydrates compared to the present findings4<\/sup>. D. alata<\/em> tubers are considered energy-giving\nfood crops due to their appreciable amount of carbohydrates. Starch is the most\nabundant form of carbohydrates which store energy in plants. Another study reported\na much higher value of starch (62.94%) than the present study.34<\/sup> Fat supplies more\nthan twice the energy furnished by carbohydrates or protein per unit weight.\nFat in the diet helps with the absorption of fat-soluble vitamins, and it also\ncontributes to the palatability of food.29<\/sup> Fat is an essential component of the diet. An\nearlier study reported 1.62% of fat and 8.40% of protein for Dioscorea alata<\/em>.32<\/sup> Both areal and underground tuber\ncontained appreciable amounts of vitamin C, which indicates consumption of this\ntuber could help in the absorption of iron, and decrease atherosclerosis and\nsome kind of cancer.35<\/sup> Minerals are inorganic nutrients only needed in small\nquantities.36<\/sup>\nMinerals are necessary for the bulk of the body’s metabolic processes\u2014electrolytes\nlike potassium and salt help keep fluid and blood volume in check. Blood\npressure rises when people consume too little potassium and too much sodium.\nPhosphorus is needed for various functions, including ATP generation, signal\ntransmission, and bone mineralisation. Iron is a component of cytochromes and electron\ntransport. Available literature shows that D.\nalata<\/em> is a good source of minerals. 4,31,32<\/sup><\/p>\n\n\n\n

Bioactive compounds<\/strong><\/p>\n\n\n\n

Bioactive compounds are biologically\nactive substances which have positive or negative effects on living\norganisms.37<\/sup>\nFlavonoids are a class of polyphenolic chemicals with several benzene rings.\nAllergies, inflammation, free radicals, platelet aggregation, bacteria, ulcers,\nhepatotoxins, viruses, and cancers are all protected by flavonoids.38<\/sup> Phenolic compounds\nare potent antioxidant and scavenging agents. The present study suggested that D. alata<\/em> is a rich source of phenols and flavonoid content.\nThis report was corroborated by other studies 9,14,25. Dioscorea species are an\nessential source of Diosgenin, a commercially vital bioactive sapogenin.39<\/sup> It is used to\nmanufacture crucial pharmaceutical steroidal drugs, such as precursors, to\nproduce sex hormones and oral contraceptives.39<\/sup> Tannin is one of the phenolic compounds\nwhich give an astringent and bitter taste.38<\/sup> A previous study reported 0.58mg\/100gm of\ntannin for D. alata<\/em>.34<\/sup>    <\/p>\n\n\n\n

DPPH scavenging activity<\/strong><\/p>\n\n\n\n

Antioxidant\nmolecules in food are a vital protective factor for one’s health. Antioxidants\nprotect by these defence methods: the first line of defence prevents excessive\nformation of reactive oxygen species by inactivating endogenous cations like\nFe+ and Cu+. The second line of defence comprises tocopherols, tocotrienols,\ncarotenoids, ascorbic acid, and other phytochemicals that can scavenge reactive\noxygen species. The result\nof the present study showed that the underground tuber has more DPPH scavenging\npotential than the areal tuber. Similarly, a study from India also recorded\nunderground tuber of D. alata<\/em> is a more potent DPPH scavenger than the aerial\ntuber.14<\/sup> D. alata<\/em> tuber exhibited\nDPPH scavenging potential might be due to the presence of phenols, flavonoid,\ntannin, diosgenin and ascorbic acid in tuber 6, 9, 35, 40<\/sup>.            <\/p>\n\n\n\n

Antimicrobial activity <\/strong><\/p>\n\n\n\n

Based on susceptibility tests that produce MIC in 100-1000 mg\/mL, phytochemicals are classified as to whether they have antimicrobials.41<\/sup> If the MIC values are observed below 100 \u00b5g\/mL, the activity is considered significant and moderate when 100<MIC<625 \u03bcg\/mL.42,43<\/sup> The agar healthy diffusion test against all tested clinical strains revealed that the crude extracts from D. alata<\/em> showed significant to moderate antibacterial activity. Therefore, the activity recorded with the natural section on the clinical stress of A. baumannii, E. faecalis, K. pneumonia, P. mirabilis, P. aeruginosa, S. aureus <\/em>(MRSA), S. pyogenes<\/em> has similar activity with the earlier report 2,8,9,44<\/sup>. A study reported an inhibition zone of 12mm against Salmonella paratyphi and Shigella dysenteriae<\/em> for chloroform soluble fraction of D. alata<\/em> at a concentration of 400\u00b5g\/disc8<\/sup>.  Another study recorded an inhibition zone of 1cm against S. pyogenes<\/em> at a concentration of 1mg\/ml for acetone extract of D. alata<\/em>.2<\/sup> At the same time, tuber extract of 500\u00b5g\/ disc showed a maximum inhibition zone of 17.16mm against Shigella dysenteria9<\/sup>. D. alata tuber exhibited antibacterial activity may be due to the presence of phytochemicalsPhenol, flavonoid and tannin in the tuber.25,45-47<\/sup> The tannin inhibits the synthesis of cell protein in bacteria because tannin forms irreversible complexes with proline-rich proteins.45<\/sup> Carbonyl group of flavonoids form complexes with extracellular and soluble proteins within the bacterial cell wall.46,47<\/sup> <\/p>\n\n\n\n

Toxicity\nanalysis <\/strong><\/p>\n\n\n\n

Administration\nof doses up to 8000 mg\/kg body weight of D.\nalata <\/em>methanolic extract did not reveal any behavioural changes or\nmortality in rats during the entire observation period. Therefore, LD50<\/sub>\nof D. alata <\/em>methanolic section may be\ngreater than 8000 mg\/kg. Daily doses of methanolic extract of D. alata<\/em> for up to 45 days failed to\nreveal any significant difference in various blood biochemical parameters\nbetween treated and untreated groups, indicating no side effects to animals.\nNecrosis, infiltration, oedema and conjunction, which are a sign of\nhepatotoxicity, were not observed in the liver cells of the experimental group.\nThe kidneys revealed normal glomeruli, proximal and distal tubules,\ninterstitium, and blood vessels. The result of the study indicated that the consumption\nof D. alata<\/em> is entirely safe. <\/p>\n\n\n\n

 Conclusion<\/strong><\/p>\n\n\n\n

The\nnutritional composition, antioxidant and antimicrobial activity of the\nunderground and aerial tuber of D. alata<\/em> were evaluated in this present\nstudy. Results of the analysis suggested that the nutritional composition of\nthe underground tuber is more vibrant than the aerial tubers. Further, the\nantioxidant activity of the underground tuber was found to be significantly\nvery high compared to the aerial tuber. The present study also emphasizes the\nphytochemical analysis and antimicrobial potential of D. alata<\/em> against\nclinical microbial cultures. The extracts of D. alata<\/em> tubers have shown\nexcellent activity against A.\nbaumannii, E. faecalis, K. pneumonia, P. mirabilis, P. aeruginosa, S. aureus <\/em>(MRSA) and S. pyogenes<\/em>. Hence,\nthe tuber D. alata<\/em> can be used for functional\nfood and as a potential antimicrobial agent against pathogenic microorganisms. All\nthe biochemical parameters were found to be within the normal range in all the\ntreated groups, and also there were no changes in the structure of the kidney\nand liver in all the treated groups. The raw and boiled D. alata does <\/em>not create any injury. <\/em>There was an absence of acute and sub-acute toxicity in mice.<\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

None<\/p>\n\n\n\n

Conflicts of Interests<\/strong><\/p>\n\n\n\n

The authors do not have any conflicts of interests<\/p>\n\n\n\n

Funding Sources<\/strong><\/p>\n\n\n\n

None<\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

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