Indexing links of GJRMI

GJRMI has been indexed in the Following International Databases

Google Scholar, ProQuest, DHARA online; DOAJ; Index Copernicus; NewJour; ScienceCentral;

getCITED; RoMEO; Geneva Foundation for Medical Education & Research ; Catalog ebiblioteca;

Ayurbhishak; Medicinal plants (Dravya Guna); Indianscience.in; Necker; Hong Kong University

of Science and Technology Library; University of Zurich; University of Kansas; Western

Theological Seminary; CaRLO; Mercyhurst University; University Library of Regensberg; WZB;

Jadoun science; University of California, San Fransisco (UCSF Library); University of

Washington; University of Saskatchewan; University of Winnipeg; Universal Impact Factor;

Global Impact factor, Ulrich’s Periodicals Directory, New York Public Library, WISE, Cite factor,

DRJI, Miami University Libraries,

AYUSH RESEARCH PORTAL - Department of AYUSH, Ministry of Health & Family welfare,

Govt. of India

-

All types of Keraliya Ayurvedic treatments available for all the diseases)

Ayurvedic Treatments in the following diseases: Eye diseases, Asthma, Skin diseases, Joint

diseases, Diseases of the nervous system, Gynaecological & Obstetric diseases, Obesity, Asthma, Stress,

Anxiety, Insomnia, Depression, Loss of Memory & Concentration, Piles, digestive tract diseases,

Infertility etc.

Address: No. 40, IInd cross, KV Pai Layout, Konanakunte,

Near Silicon city school, Bangalore – 62, Karnataka, India.

Contact: Mobile: +919480748861

Chakradatta Ayurveda Chikitsalaya, Mysore. (Panchakarma & Netra Roga Chikitsa Kendra)

Consultant Physician: Dr. Ravi Kumar. M.

(Specialized in different types of Keraliya Ayurvedic treatments especially in ENT & Eye diseases)

Get treated through Ayurveda, at our Hospital. (Exclusive Panchakarma Therapy available with accommodation)

Address: Beside Vikram Jyothi Hospital, Temple Road, V V Mohalla,

Mysore – 12, Karnataka, India.

Contact: Mobile: +919980952358, +919035087999

E- mail: raviamrita.kumar9@gmail.com

Arudra Ayurveda, Bangalore

(A PANCHAKARMA TREATMENT CENTRE)

An International, Peer Reviewed, Open access, Monthly E-Journal

ISSN 2277 – 4289 www.gjrmi.com

Editor-in-chief

Dr Hari Venkatesh K Rajaraman

Managing Editor

Dr. Shwetha Hari

Administrator & Associate Editor

Miss. Shyamala Rupavahini

Advisory Board

Prof. Rabinarayan Acharya Dr. Dinesh Katoch

Dr. S.N.Murthy Dr. Mathew Dan Mr. Tanay Bose

Dr. Nagaraja T. M. Prof. Sanjaya. K. S. Dr. Narappa Reddy

Editorial board

Dr. Kumaraswamy Dr. Madhu .K.P

Dr. Sushrutha .C.K Dr. Ashok B.K.

Dr. Janardhana.V.Hebbar Dr. Vidhya Priya Dharshini. K. R.

Mr. R. Giridharan Mr. Sriram Sridharan

Honorary Members - Editorial Board

Dr Farhad Mirzaei Mr. Harshal Ashok Pawar

Dr. Sabarinath Subramaniam Dr. Yogitha Bali

INDEX – GJRMI - Volume 3, Issue 10, October 2014

MEDICINAL PLANTS RESEARCH

Biology

CULTIVATION OF OYSTER MUSHROOM (PLEUROTUS OSTREATUS) ON WASTE PAPER

WITH SUPPLEMENT OF WHEAT BRAN

Asefa Keneni, Geda Kebede 370–380

Pharmacology

WOUND HEALING ACTIVITY OF ALOCASIA MACRORRHIZOS (L.) G.Don PLANT – AN

EXPERIMENTAL STUDY

Santosh kumar singh, Sonia Thakur, Neha Shukla, Sanju Singh 381–388

COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – FRUIT OF ATI BALA – ABUTILON INDICUM (L.) SWEET. OF THE

FAMILY MALVACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT,

KARNATAKA, INDIA

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal

CULTIVATION OF OYSTER MUSHROOM (PLEUROTUS OSTREATUS) ON

WASTE PAPER WITH SUPPLEMENT OF WHEAT BRAN

Asefa Keneni1*, Geda Kebede

2

1,2Department of Biology, College of Natural and Computational Sciences, Ambo University, Ethiopia.

*Corresponding author: E-mail: asefa_keneni@yahoo.com

Received: 16/08/2014; Revised: 28/09/2014; Accepted: 30/10/2014

ABSTRACT

The present study was under taken to evaluate the usability of waste paper as a major substrate

with supplement of different ratio of wheat bran for cultivation of oyster mushroom. Five different

treatments (T1-T5) were used in the study and these treatments showed significant (P≤0.05)

variation. T2 showed the fastest mycelial extension (0.26 cm/day) and T4 and T5 showed slowest

mycelial extension (0.14 and 0.15 cm/day). T2 and T3 showed shortest incubation periods (85 days)

and T5 had longer (105 days) for overall cycle of the mushroom production. T4 showed shortest

mean periods from pinning to maturation in the 2nd

, 3rd

and 4th

harvests (8 to 5 days), while T1 took

longer incubation periods 10 to 8 days in all the three harvests. T3 showed highest fresh weight in 1st

flush (900g) and T5 gave least fresh weight (150g). In T2 and T3 the forth harvest was lowest (130

and 124g) as compared to the former harvest while in the rest of the treatments it was absent.

Maximum number (12) of bunches was recorded on T2 and the least on T5 (3). Pilus diameter was

maximum from T3 (14 cm) and the minimum (8cm) was noticed from T5. However the stipe length

of the mushroom from the different treatments did not vary considerably (2.5–3.0cm). The highest

numbers of fruiting bodies were collected from T2 and T3 (72) and the least from T5 (15). Higher

number of aborts was recorded on T2 (110) and the lowest on T5 (20). The highest total wet/fresh

weight of matures and biological efficiency were recorded in T2, and T3 (2090–2214g, 104–110%

respectively) and the least from T5 (370g, 6% respectively). The results obtained indicate that

supplement of different ratio of wheat bran significantly resulted in the variability of growth, yield

and biological efficiency of oyster mushroom and T3 and T2 may be used for commercial production

of oyster mushroom in the areas where other substrates may be a limiting factor.

KEY WORDS: Biological efficiency, Wheat bran, Oyster mushroom, Waste paper

Research Article

Cite this article:

Asefa Keneni, Geda Kebede (2014), CULTIVATION OF OYSTER MUSHROOM

(PLEUROTUS OSTREATUS) ON WASTE PAPER WITH SUPPLEMENT OF WHEAT BRAN,

Global J Res. Med. Plants & Indigen. Med., Volume 3(10): 370–380

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

INTRODUCTION

Mushrooms are the fungi that have been used as food since time immemorial. Nutritionally they are a valuable source of health food, which is low in calories, and rich in carbohydrates, essential amino acids, fibre, important vitamins and minerals. Mushrooms have also been used in medicine for centuries in the Orient but their potential as health potentiators and elicitors of immune system is recent (Chang and Miles, 1989; Synytsya et al., 2008).

In addition, mushroom cultivation is considered as a possible option to alleviate poverty and develop the life style of the vulnerable people. In addition mushroom cultivation offers benefit to market garden when it is integrated in to the existing production system by producing nutritious food at a profit, while using materials that would otherwise be considered “waste” (Beetz and Kustida 2004; Sharma et al., 2013). This is because mushroom contains many essentials nutrients and they are found to solve dietary related health problems (Synytsya et al., 2008).

Mushrooms are eaten as meat substitutes and flavoring agents. In addition to the nutritional and medicinal values, mushroom cultivation practices have a paramount importance in food self sufficiency; especially for low income country’s like Ethiopia. Mushroom can generate additional trade offering opportunities through processing enterprises. Mushroom cultivation is suitable for all job seeking groups including elders, disabled and youngsters. Besides, mushroom cultivation is labor intensive and creates job opportunities (Dawit, 1998). It also derives toward full uses of all materials in which nothing is left as waste, without any adverse impacts on the environment through sustainable utilization of lignocelluloses wastes available usually as byproducts form agriculture, forestry and households (Chang and Miles, 1989). Currently, mushroom are regarded as the most profitable and environment friendly method for recycling of the vast lignocelluloses waste substrates, which could otherwise be dropped

in to the environment and cause pollution (Atikpo et al., 2008).

Apart from their nutritional potentials, they are important medicinally for cholesterol reduction, immune enhancement, and cancer fighting, anti allergic activities, antimicrobial and cardiovascular treatments (Rajak et al., 2011). They also have a long history of use as traditional medicine in China. Their legendary effects on promoting good health and increasing adaptive abilities have been also supported by recent studies (Wasser, 2000). In addition to their edibility and health benefits, their mycelia can produce a group of complex extra cellular enzymes which can degrade and utilize the lignocelluloses wastes in order to reduce pollution (Atikpo et al., 2008).

The major problem associated with the transfer of technology from mushroom cultivation is lack of technical knowledge for its cultivation. Studies conducted with the relation of cultivation of mushroom indicated that agricultural residues: rice husk, sorghums Stover, saw dust, cotton seed waste, cocoa bean shell, and sawdust-Gliricidia mixture were found to be suitable substrates (Rajak et al., 2011).

Despite high diversity of wild edible mushrooms has been there in Africa especially in Ethiopia, its recognition very little. Cultivation and production of mushroom has not been practiced on commercial scale in most developing countries which has consequently affected commercial mushroom marketing which is yet to be embraced by most farmers (Abate, 1998; Atikpo et al., 2008).

In developing countries, governmental and non-governmental organizations have not given due attention to mushrooms as an important crop that can fetch farmers substantial income to alleviate poverty (Atikpo et al., 2008). Similarly it is accepted that mushroom are not a luxury food but a national necessity to combat poverty and malnutrition (Chang, 2008). However, there is no mushroom cultivation practice in the country to fill the demands of people in tested in the mushroom consumption. Those very few mushroom farmers in Ethiopia

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

are restricted to the capital city. Some research based practices in some parts of the country are still at the stage of trials.

From all edible cultivated mushrooms; oyster mushroom (Pleurotus ostreatus) is considered as versatile fungal strain in its ability to use various organic waste materials to grow and give releasable yield. Besides, this mushroom strain is easier to grow by the beginner and also in a wide range of environmental conditions. Large amount of used waste papers has been produced from different governmental and non-governmental institutions as well as a pack for different goods. The well known methods of removal or reduction of these waste papers from the given area was by burning it. Burning of solid organic wastes including waste paper increases the emission of green house gases and not environmental friendly. Cultivation of mushroom on waste paper with supplement of different proportion of wheat bran was not reported from Ethiopia. Mintesnot et al. (2013) evaluated biomass of some invasive weed species as substrate for oyster mushroom and observed maximum biological efficiency on Parthenium hysterophorus; Beje Gume et al. (2013) evaluated of eight locally available substrates and substrate combinations for their productivity and biological efficiency (BE) for cultivation of commercial mushroom strain (Pleurotus ostreatus) and observed that the fastest mean value (0.69 cm/day) of mycelial extension was recorded from sdZcCh (combination of sawdust of Cordia africana and Pouteria adolfi-friederici, corncobs and coffee bean husks). However, mycelial growth in coffee bean husks was completely ceased after 15 days. The present study was undertaken mainly to assess the growth, yield and biological efficiency of oyster mushroom on substrates composed of different proportion of waste paper and wheat bran mixed on dry weight basis.

MATERIALS AND METHODS

Organism and culture conditions

The fungal strain, Pleurotus ostreatus (Oyster mushroom) was obtained from

Mycology Laboratory, Department of Biology, Addis Ababa University, Addis Ababa, Ethiopia. The pure culture of Pleurotus ostreatus was transferred on to Potato Dextrose Agar (PDA) prepared in the laboratory using fresh potato 250 g; glucose (Dextrose) 20 g; agar 20 g and chloramphenicol 0.2 g in 1000 ml of water. The medium was poured into the Petri dishes and allowed to cool in under aseptic condition in laminar flow chamber. The cooled and solidified medium was inoculated by 1 cm×1 cm agar block of the fungal strain

and incubated at 25 C. The growth of the culture and presence of contamination were visually inspected at three days interval.

Grain Spawn production

In this study, the spawn (mushroom seed) of Pleurotus ostreatus was produced on yellow colored sorghum grain, wheat bran and calcium sulfate (gypsum) in the ratio of 88:10:2 respectively (Dawit, 1998). The required amount of sorghum grain was weighed and soaked over night in sufficient amount of water. The grains were washed and drained to remove the dead and floating seeds with excess of water. After removing the excess water from the grain, the required amount of wheat bran and gypsum (CaSO4 2H20) were added and transferred to 1000 ml glass bottles (75% level) leaving a head space over the grain and autoclaved at 121°C temperature for 45 minutes. After cooling, each bottle was inoculated with 20 agar blocks (1 cm × 1 cm) of 15day old mushroom culture from the Petri

dish and incubated for 21 days at 28 ± 2 C until the substrate were fully colonized and the mycelia invasion and contamination were inspected at five days interval.

Waste paper was collected from different departments of the University and wheat bran was collected from the local market.

Treatments

Five treatments (T1–T5) comprising different proportions of waste paper and wheat bran (2000 g) along with lime stone (Calcium Carbonate 20 g) on dry weight basis were used as shown in Table 1.

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Table1: The composition of different treatments

Treatment waste paper

(g)

Wheat bran

(g)

Total (g)

T1 1800 200 2000

T2 1600 400 2000

T3 1400 600 2000

T4 1200 800 2000

T5 1000 1000 2000

Preparation of the substrate

The waste paper was cut into small pieces approximately (3–5 cm), weighed and soaked in sufficient amount of water immediately before use. Excess water present in the substrates was drained thoroughly and mixed with required amount of wheat bran and one percent calcium carbonate and filled in sterilizable yellow color polyethylene bags (Kurtu pestal). The substrates were autoclaved

at 15Psi pressure at 121 C temperatures for 1h. After sterilization the substrates were transferred to transparent polyethylene cultivation bags for easy supervision of the growth of the mycelia and presence of contamination. Each substrate (2,000 g) with 70% moisture was mixed with 10% spawn (dry weight/wet weight basis) and the inoculated polythene bags were then tightly tied with string made from polyester/cotton cloth. Pin holes were made through the bags (1/100 cm

2)

for drainage and aeration. It was kept in a spawn running room at room temperature in the dark until primordia were formed. After primordial formation, large holes were made in the polythene bag to allow normal development of fruiting bodies. Bags were transferred to mushroom house under normal environmental conditions and relative humidity (the room maintained at 85–90%) by keeping water in open containers at different corners of the room. The cultivation bags were irrigated using tap water every morning and evening until all flushes of Pleurotus ostreatus fruiting bodies were harvested. Adequate ventilation was provided to prevent increased CO2 concentration in the room by opening the door and windows of the room for half an hour in the morning and in the evening. The

mushrooms were manually harvested at maturity which was indicated by up ward curving of the edges of the cap.

Biological efficiency was calculated and defined as the ratio of weight (g) of fresh mushrooms harvested to dry weight (g) of the substrate. Biological Efficiency = Weight of fresh fruiting bodies (g) × 100 Weight of dry substrate (g)

Data analysis

The data were analyzed by comparing the mean weights and percent biological efficiency through one way ANOVA. The data groups were analyzed using Statistical Package for Social Sciences (SPSS) for windows 16.0. Treatment mean were compared using LSD.

RESULTS

Mycelia extension

There were significant (P≤0.05) differences in the mycelial extension of oyster mushroom grown on different substrates. T2 showed the fastest mycelial extension followed by T3 while, T4 and T5 exhibited slowest mycelial extension on 7

th and 14

th days of incubation

periods (Table 2). There were significant (P≤0.05) differences in the days required for complete invasion of the substrates receiving different treatments. The time required for complete invasion of the substrates was significantly (P≤0.05) less for T2 and T3 when compared to that of T1 and T5 (Table 2). Total days required to complete the production cycle was observed shortest for T2 and T3 while it took more days for T5.

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Table 2: Mycelial extension on the substrates at different treatments measured on 7th

and 14th

days of incubation

Treatments Mycelia extension in (cm) Mean values

(cm/day)

Number of days

required for

complete invasion

Total days

required to

complete the cycle 7

th day 14

th day

T1 1.9 5.5 0.16 26 90

T2 2.5 6.5 0.26 22 85

T3 2.3 6.25 0.24 23 85

T4 1.2 4.5 0.14 25 95

T5 1.1 4.3 0.15 27 105

Growth rate of mushroom (Flushes)

Mean incubation periods of mushroom

flushes showed highly significant differences

(P≤0.05). T1 showed relatively shorter

incubation to 1st flush while 1

st–2

nd flush and

2nd

–3rd

flush took more days than T2 and T3.

T2 and T3 took moderate days from

incubation to 1st flush and shortest for the

remaining three consecutive harvests as

compared to the other treatments. T5 took

longer incubation periods at all harvest.

Besides number of harvest significantly varied

for different treatments; T2, T3 and T4 gave

four harvests; T1 three harvests and T5 only

two harvests (Table 3).

Pinning to maturation duration of oyster

mushroom

The mean periods taken from pinning to

maturation of each treatment showed

significant (P≤0.05) variation. T1 and T5

relatively took longer periods from pinning to

maturation and T2, T3 and T4 took shorter

periods from pinning to maturation in all

flushes as compared to other treatments (Table

4).

Yield of mushroom per flushes

Yield of mushroom per flush (wet weight)

showed significant variation between

treatments (P≤0.05) (Table 5) as well as

between flushes. T3 showed highest fresh

weight in grams in 1st

and 2nd

flushes

followed by T2; the result observed with T1

and T4 were not comparable with the highest

yielding treatments, while T5 was poor in this

regard. In 3rd

flush, T2 and T3 gave relatively

higher fresh weight of mushroom while all the

remaining treatments showed least. In all the

treatments the 4th

flush not comparable with

other harvest cycle (Table 5).

Number of bunches, matures and aborts

More number of bunches were recorded on

T2 followed by T3 while all the remaining

treatments showed least in number of bunches.

The highest and equal numbers of fruiting

bodies were collected from T2 and T3. T5 and

T4 gave the least number of fruiting bodies.

Higher number of aborts were recorded with

treatments T2 followed by T3 and the least

number of aborts were recorded in T1 (Fig1).

Table 3: Incubation periods of different harvests

Treatments Incubation -1st flush 1

st–2

nd flush 2

nd–3

rd flush 3

rd–4

th flush

T1 40 18 16 −

T2 45 15 13 12

T3 45 15 23 12

T4 50 17 15 13

T5 55 18 − −

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Table 4: Pinning to maturation of the oyster mushroom under different treatments regimes

Treatments Mean duration (days)

1stFlush 2

nd Flush 3

nd Flush 4

nd Flush

T1 10 9 9 −

T2 8 7 6 6

T3 8 7 6 6

T4 8 7 6 6

T5 9 8 − −

Mean values with in a column sharing the same superscript letter(s) are not significantly different by using LSD test at

P≤0.05

Table 5: Mean yield per flush in the different treatments

Treatments Mean fresh- weight of mushroom (g)

1stFlush 2

nd Flush 3

nd Flush 4

nd Flush Total

T1 413 251 150 − 814

T2 880 560 520 130 2090

T3 900 765 515 125 2215

T4 290 210 150 75 560

T5 150 130 90 − 370 Mean values with in a column sharing the same superscript letter(s) are not significantly different by using LSD test at

P≤0.05

Fig 1: The different stages of mushroom production in this experiment

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Pilus Diameter and Stipe length

Pilus diameter was found to be the largest

for the samples collected from T3 followed by

T2, T4 and T1 respectively, while it was

smallest for the sample collected from T5. The

stipe length of the samples collected from

different treatments did not show significant

variation (Fig2).

Total yield and Biological efficiency

The highest total wet/fresh weight of

matures was recorded in T3, followed by T2.

The least total fresh/wet weight was recorded in

T5 (Table 5). The effect of different treatments

on biological efficiency of oyster mushroom

showed significant (P≤0.05) differences. The

highest biological efficiency was recorded with

T3 followed with T2. The least was recorded

with the treatment T5 (Fig 3).

Fig 2: Number of bunches, matures and aborts of different treatments

Fig 3: Pilus diamter and stipe lengths of different treatmnets

0

20

40

60

80

100

120

1 2 3 4 5

Nu

mb

er

of

bu

nch

es,

mat

ure

s an

d a

bo

rts

Treatments

Number of bunches

Number of matures

Number of Aborts

0

2

4

6

8

10

12

14

16

1 2 3 4 5

Pilu

s D

iam

ete

r an

d S

tip

e le

ngt

h(c

m)

Treatments

Pilus Diameter

Stipe length

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Fig4: The biological efficiency of different treatments

DISCUSSION

Selection and optimization of available

substrates in order to obtain reasonable yield of

mushroom could be considered as a priority

target in mushroom research. The use of waste

paper as a major substrate for the cultivation of

oyster mushroom was not yet practiced in

Ethiopia. In this experiment the complete

colonization of the substrate took 22 days in the

faster and 27 in the slower and then another 14

days were taken after complete invasion of the

mycelium to first harvest in the fastest

treatment and 28 days in the slower. In this

study, the periods taken for spawn running on

the different treatments showed relatively

longer time as compared to results reported in

the literature. Ashraf et al. (2013) reported

shorter periods for the substrate completely

colonized by mycelium of the different oyster

species which also indicates differences among

the different species and the substrates.

According to these authors the minimum

number of days 16.20 took by P. ostreatus

16.20 ± 0.59 while species P. sajor-caju and P.

djmor showed same level of significance with

18.07 ± 0.69 and 18.67 ± 0.61. Oseni et al.

(2012) reported periods of colonization to first

harvest from 33 to 43 days on fermented saw

dust supplemented with different proportions of

wheat bran. In this study the different

treatments showed significant variation periods

taken for primordial formation after the

complete colonization of the substrate by the

fungal strain. T1 took only four days for

initiation for the primordial formation while T5

took another 19 days for initiation for the

primordial formation. These longer days of

initiation of primordial formatyion after

mycelia running may be due to slow releasing

of nutrients from waste paper as compared to

other substrates, for example, wheat straw and

rice straw on which much of research work has

been done on this mushroom species. Ashraf et

al.(2013) reported that all the treatments they

tested showed 3.73 to 5.13 days for primordial

initiation after mycelia running.

In this study, in all the treatments, the

successive pinning to harvest duration was

shortened by at least a day. The shortest mean

duration of pinning to maturation was 8 in the

1st, 7 in the 2

nd, 6 in 3

rd and 4

th. And the longest

mean duration of pinning to maturation was 10,

in the 1st, 9 in the 2

nd and 3

rd. The duration

observed in the present study was longer when

compared with the reports in the literature

(Gume et al. (2013) which was 3.3 in the

shortest and 6.0 in the longest. Studies

indicated that environmental factor affects the

incubation periods of oyster mushroom.

According to Zadrazil (1976) and Daba et al.

(2008) longer period of incubation for oyster

0

20

40

60

80

100

120

1 2 3 4 5

Bio

logi

cal E

ffic

ien

cy(%

)

Treatments

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

mushrooms was at lower temperatures and low

relative humidity.

In this study, in all the treatments the yield

(fresh/wet weight) of the mushroom harvested

in the first cycle was greater than the remaining

successive harvests. In addition to the

minimized yield in the next consecutive

harvest, treatment T1 and T5 did not give

harvest at the fourth cycle. Our observation on

the different harvest is in line with reports in

the literature. Ashraf et al. (2013) reported that

the different treatments vary in the amount of

mushroom yield harvest at different flushes and

at each successive harvest, the amount of the

yield declined. Number of bunches formed on

different treatments were significantly

different. T2 produced highest number of

bunches, followed by T3 and all the remaining

treatments gave least number of bunches. The

highest and equal numbers of fruiting bodies

were collected from T2 and T3 (72). T5 (15)

and T4 (22) gave the least number of fruiting

bodies. Higher number of aborts observed in

treatments T2 (110) and T3 (92) and the least

number of aborts T1(16), this may be due to

the optimal proportion of waste paper and

wheat bran which allowed maximum

primordial formation from which some of them

aborted. In this study more number of bunches

result in more number of fruiting bodies. This

observation was in line with the results reported

by Gume et al. (2013) who reported that

substrates that gave higher yield also contained

higher number of propagating fruit bodies per

bunch and highest variability among different

treatments on the mean number of mature fruit

bodies and aborts. In majority of the substrates,

the number of pinhead abortions exceeded

number of matures. Kimenju et al. (2009)

reported that more than 50% of pinheads

emerged did not grow into marketable

products. Gume et al. (2013) observed high rate

of pinhead abortion from low-yield substrates

such as sd1C and ZcCh. The largest pilus

diameter was measured with T (14 cm) and the

smallest with T5 (8 cm); the rest of the

treatments gave pilus diameter between the

largest and the smallest. Largest pilus diameter

significantly increased the total fresh/wet

weight of oyster mushroom. Oseni et al. (2012)

reported highest mean pilus diameter 57.9 to

62.3 mm on sawdust supplemented with

different levels of wheat bran. The largest pilus

was obtained from sawdust substrate

supplemented with 15% wheat bran (62.3 mm)

and the smallest obtained on sawdust substrate

supplemented with 5% wheat bran (57.9 mm).

The pilus diameter obtained in the present

study was greater than all those reported

earlier, may be due to the varied proportions of

the major substrate (waste paper) which can

supplement the necessary nutrients for

mushroom growth.

The stipe length of all the 5 treatments did

not vary significantly (2.5–3.0 cm), which is in

agreement with the results of Gume et al.

(2013). 1.4–1.9 cm. Oseni et al. (2012)

observed stipe length of oyster mushrooms

ranging from 39.4–59.5 mm (3.94–5.95cm) on

fermented sawdust substrate supplemented with

different wheat bran levels and highest stipe

length (59.5 mm) (5.95 cm) was observed on

substratum supplemented with 15% wheat

bran. The mean number of mature fruit body

and aborts were greatly varied among the

different treatments in the study. The total fresh

weight of the mushroom was highest in T3

followed by T2 (2214–2090 g per 2000 g of the

dry substrate and their biological efficiency

(110–104%). In all the parameter tested T3 and

T2 (1400 g waste paper 600 g wheat bran and

1600 g waste paper 400g wheat bran) found to

superior this may be due the proportion of the

waste paper and wheat bran for bio-availability

of the various nutrients contained in the

substrates mixture. In this study the least total

fresh/wet weight of the mushroom and

biological efficiency was recorded in T5 (370 g

per 2000 g dry substrate and 6% BE. In all the

treatments yield of mushroom declined

successively throughout the four cropping

periods. Kimenju et al. (2009) reported that

yields of mushroom in different substrates

slightly declined from the first flush to the

successive harvests. The crops of oyster

mushroom were harvested in four flushes and

the maximum yield was obtained in the first

flush than the 2nd

, 3rd

and 4th

flushes,

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

respectively as observed by Oseni, et al.

(2012).

CONCLUSION

Production of edible mushroom has been

considered as diversification of food production

and also contribute in the struggle for food self

sufficiency and attaining food security

particularly in the developing world like

Ethiopia. Testing the usability of waste paper

as a major substrate with the supplement of

different ratio of wheat bran was not yet tried

for mushroom production in Ethiopia. The

different treatments resulted in significant

variation on growth, yield, yield parameters

and biological efficiency of oyster mushroom.

From all the treatments T3 and T2 observed to

be considered as highest yielding with all the

parameters tested and recommended for

commercial production of oyster mushroom.

While the rest of the treatments performed

below the acceptable yield and biological

efficiency. The result of this study shows the

possibility of mixing waste paper with wheat

bran in different proportion and obtaining high

yield and good quality mushroom fruiting

bodies. The future research direction should

focus on developing a substrate that will give

highest yield of mushroom by mixing waste

paper with other locally available organic

wastes.

ACKNOWLEDGEMENT

The authors are greatly acknowledging

Ambo University, Ethiopia for extending the

financial support for this research work.

REFERENCES

Ashraf J, Asif Ali M, Ahmad W,Muhammad

Ayyub C, Shafi J (2013). Effect of

Different Substrate Supplements on

Oyster Mushroom (Pleurotus spp.)

Production Food Science and

Technology 1(3): 44–51,

Atikpo M, Onokpise O, Abazinge M, Louime

C, Dzomeku M, Boateng L Awumbilla

(2008). Sustainable mushroom

production in Africa A case study in

Ghana. Afr. J. Biotechnol., 7: 249–253.

Beetz, A. and M. Kustida, (2004). Mushroom

Cultivation and Marketing. ATTRA

Publication # IP 087, Retrieved from:

http://attra.ncat.org/attrapub/

mushroom.html.

Beje Gume, Diriba Mulata and Dawit Abate

(2013). Evaluation of locally available

substrates for cultivation of oyster

mushroom (Pleurotus ostreatus) in

Jimma, Ethiopia. African Journal of

Microbiology Research.,7(20): 2228–

2237

Chang, S.T. AND P.G. Miles, 1989, Edible

mushrooms and their cultivation vol.1,

CRC Press Boca Raton, FL; USA,

ISBN-13: 9780849367588, PAGES:

345.

Chang, S.T., 2008. Overview of Mushroom

Cultivation and Utilization as

Functional Foods. Retrieved from:

http:// media. wiley. com/ product_

data/

excerpt/69/04700540/0470054069. pdf,

(Accessed on: January 19, 2009).

Daba AS, Kabeil SS, Botros WA, El-Saadani

MA (2008). Production of mushroom

(Pleurotus ostreatus) in Egypt as a

source of nutritional and medicinal

food. World J. Agric. Sci., 4:630–634.

Dawit A (1998). Mushroom Cultivation: A

practical approach, Berhanena Selam

Printing Enterprise, Addis Ababa

Ethiopia.

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 370–380

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Kimenju JW, Odero OM, Mutitu EW, Wachra

PM, Narla RD, Muiru WM (2009).

Suitability of locally available

substrates for oyster mushroom

(Pleurotus ostreatus) cultivation in

Kenya. Asian J. Plant Sci., 8:510–514.

Lqbal SM, Rauf CA,Sheikh M (2005). Yield

performance of oyster mushroom on

different substrates. Int.J.Agric bio.7

:900–903.

Mintesnot B, Ayalew A, Kebede A (2013).

Evaluation of Some invasive weed

species as substrate for oyster

mushroom (Pleurotus spp.) cultivation.

Pak. J. Biol. Sci. _: 1-7 DOI 10.3923

Oseni T O, Dube S S, Wahome, P K,

Masarirambi, M T, and Earnshaw D M

(2012). Effect of Wheat Bran

Supplement on Growth and Yield of

Oyster Mushroom (Pleurotus

Ostreatus) on Fermented Pine Sawdust

Substrate. Experimental Agriculture &

Horticulture :V-30–40

Park, G. and Kwang, H.O. (2001).Nutritional

value of a variety of

Mushrooms.www.Mushworld.com/sub-

en.html.

Pathmashini L, Arulnandhy V, Wijeratnam SW

(2008) cultivation of oyster mushroom

(Pleurotus ostreatus ) on saw dust J.

Biol. Sci., 37: 177–182.

Rajak S, Mahapatra S.C. and Basu M 2011.

Yield , Fruit body diameter and

cropping duration of oyster mushroom (

Pleurotus sajor caju ) grown on

different grasses and paddy straw as

substrate. European Journal of

Medicinal plants. 1(1): 10–17.

S.R mondal, M.J. Rehana, M.S. noman and

S.K.Adhikary 2010. Comparative study

on yield performance of oyster

mushroom (Pleurotus florida) on

different substrates Agrotechnology

discipline, Khulna University, Khulna.

9208, Bangladesh.

Sharma S, Kailash R, Yadav P., Pokhre C P

(2013). Growth and Yield of Oyster

mushroom (Pleurotus ostreatus) on

differentSubstrates. J. on New Bol.l

Rep. 2(1): 03–08

Synytsya A., Mickova, K, Jablonsky I, Slukova

M, Copikova J (2008). Mushrooms of

genus Pleurotus as a source of dietary

fibers and glucans for food

supplements. Czech. J. Food Sci.,

26:441–446.

Wasser, S. P. (2002). Nutraceuticals and bio

pharmaceuticals from edible and

medicinal mushrooms’. Int. Med.

Mushrooms. 8: 1–17.

Zadrazil F (1976). The ecology and industrial

production of Pleurotus ostreatus, P.

florida, P. cornucopiae and P. eryngii.

Mush. Sci., 9:621–652.

Source of Support: Ambo University, Ethiopia Conflict of Interest: None Declared

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 381–388

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal

WOUND HEALING ACTIVITY OF ALOCASIA MACRORRHIZOS (L.) G.Don

PLANT – AN EXPERIMENTAL STUDY

Santosh kumar singh1, Sonia Thakur

2*, Neha Shukla

3, Sanju Singh

4

1,2,3,4Assistant Professor, Mittal Institute of Pharmacy, Opp. Bhopal Memorial Hospital and Research Centre,

By Pass, Nabibagh, Bhopal- 462038, Madhya Pradesh, India.

*Corresponding Author: Email: soniathakur92@gmail.com; Mobile: +91-9425452815

Received: 18/07/2014; Revised: 28/08/2014; Accepted: 20/09/2014

ABSTRACT

A tissue injury is invariably followed by varying degree of inflammatory changes in lipid

peroxidation levels during different stages of cutaneous wound repair have been investigated earlier.

It is observed that any drug that reduces the generation of free radicals by a drug (herbal or

otherwise) during proliferative phase is a wound healing agent. The wound healing potency of

methanolic extract of leaf of Alocasia macrorrhizos was evaluated by excision, incision and

histopathological wound model on albino mice the wound healing activity was assessed for wound

contraction, period of epithelialization and skin breaking strength of granulation tissue. The result

obtained in the study revealed that methanolic leaf extract has significant wound healing potency as

compared to standard.

KEYWORDS: Wound healing, Alocasia macrorrhizos, Excision, Incision, Histopathological, etc.

Research Article

Cite this article:

Santosh kumar singh, Sonia Thakur, Neha Shukla, Sanju Singh (2014), WOUND HEALING

ACTIVITY OF ALOCASIA MACRORRHIZOS (L.) G. Don PLANT – AN EXPERIMENTAL

STUDY, Global J Res. Med. Plants & Indigen. Med., Volume 3(10): 381–388

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 381–388

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

INTRODUCTION

India has a rich flora that is widely

distributed throughout the country. Herbal

medicines have been the basis of treatment and

cure for various diseases and physiological

conditions in traditional methods practiced such

as Ayurved, Unani and Siddha. Medicinal

components from plants play an important role

in conventional as well as western medicine.

(Perumal S.R., et al., 2008, Fabricant D.S., et

al., 2001, Priya K.S. et al., 2002, Steenkamp V.

et al., 2004, Principe P., 2005).

A wound may be defined as a break in the

epithelial integrity of the skin or may also be

defined as a loss or breaking of cellular and

anatomic or functional continuity of living

tissue. According to the Wound Healing

Society, wounds are physical injuries that result

in an opening or break of the skin that cause

disturbance in the normal skin anatomy and

function. They result in the loss of continuity of

epithelium with or without the loss of

underlying connective tissue (Ramzi S.C. et al.,

1994, Strodtbeck F., 2001).

Wounds are classified as open and closed

wound on the underlying cause of wound

creation and acute and chronic wounds on the

basis of physiology of wound healing. In this

case blood escapes the body and bleeding is

clearly visible. It is further classified as: Incised

wound, Laceration or tear wound, Abrasions or

superficial wounds, Puncture wounds,

Penetration wounds and gunshot wounds

(Schultz G.S. 1999).

In closed wounds blood escapes the

circulatory system but remains in the body. It

includes Contusion or bruises, hematomas or

blood tumor, Crush injury etc.

Acute wound is a tissue injury that

normally precedes through an orderly and

timely reparative process those results in

sustained restoration of anatomic and

functional integrity. Acute wounds are usually

caused by cuts or surgical incisions and

complete the wound healing process within the

expected time frame (Lazarus G.S. et al., 1998).

Chronic wounds are wounds that have

failed to progress require a prolonged time to

heal or recur frequently. Local infection,

hypoxia, trauma, foreign bodies and systemic

problems such as diabetes mellitus,

malnutrition, immunodeficiency or medications

are the most frequent causes of chronic wounds

(Menke N.B. et al., 2007, Krishnan P., 2006).

The wound healing activities of plants have

since been explored in folklore. Many

Ayurvedic herbal plants have a very important

role in the process of wound healing. Plants are

more potent healers because they promote the

repair mechanisms in the natural way.

Extensive research has been carried out in the

area of wound healing management through

medicinal plants. Herbal medicines in wound

management involve disinfection, debridement

and providing a moist environment to

encourage the establishment of the suitable

environment for natural healing process (Purna

S.K.et al., 2000).

Alocasia macrorrhizos, (L.) G. Don.,

elephant ear Taro plant is a large evergreen,

mainly rhizomatous, sometime tuberous rooted

perennials. The plant, which belongs to the

family Araceae, is found in tropical forests and

sunny open or shaded, usually damp sites and

marshes in south East Asia. This plant is known

for many medicinal properties & hence the

present study was undertaken to study the

wound healing activity of Alocasia

macrorrhizos on excision, incision &

histopathological wound models on Albino

mice.

MATERIALS AND METHODS

Plant Aunthentication:

The leaf part of A. macrorrhizos was

collected in the month of October in Sanjivini

Ayurvedic Nursery in Bhopal, Madhya

Pradesh, India. Plants were identified by Dr.

Pradeep Tiwari, Botanist in the department of

Botany Dr. Hari Sing Gour, Vishwavidyalaya

(Sagar, Madhya Pradesh, India). A voucher

specimen has been deposited in our library for

further reference (no.1166).

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 381–388

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Preparation of leaf extract:

Fresh leaf of A. macrorrhizos were

separated from the plant and allowed to shade

dry for 15 days and then homogenized to get a

coarse powder. Powder (250 g) was extracted

with hydroalcoholic mixture (ethanol 45% and

water in 1:1 proportion) at room temperature by

cold maceration method. It was also extracted

with methanol by soxhlet method. The filterate

was collected and concentrated on a heating

mantle at 45°c till a syrupy mass was obtained.

The percentage yield was found to be 48% and

38.5% with respect to the initial dried plant

material.

Wound healing model

Selection of model:

Excision, incision and histopathological

wound model, using albino mice was selected

for assessing the wound healing activity. This

model was employed to study the rate of

wound contraction, time required for full

epithelization and tensile strength. These

parameters were selected because of easy

availability of albino mice and simplicity in

handling then.

Selection and procurement of animals:

After taking permission for animal studies,

albino mice were procured and mice of either

sex weighting 150–200gm were selected,

maintained at 24–28°C, housed individually

with free access to food and water. The animals

were left for 48hr. to acclimatize to the animal

room conditions. They were fed with standard

diet.

To perform the experiment, the mice were

divided in to three groups (n=6)

Group I – kept as control group which

received simple vehicles.

Group II – kept as which received extract of

leaf part of A. macrorrhizos formulation.

Group III – kept as standard group which

received betadine ointment.

Excision wound model:

In the excision wound model, rats were

depilated by removing hairs at the dorsal

thoracic region before wounding. Rats were

anaesthetized by diethyl ether prior to excision.

Circular wound of about 2.5 cm diameter was

made on depilated dorsal thoracic region of rats

under aseptic conditions and were observed

throughout the study. The areas of the wound

were measured (in mm2) immediately by

placing a transparent polythene graph paper

over the wound and then tracing the area of the

wound on it (approx. area 100 MM2) this was

taken an initial wound area reading.

The mice are categorized in to four groups

(n=6). The animal of group I treated as control

and only ointment base applied topically. The

animal of group II treated as standard drug and

III group treated as polyhedral applied

topically, respected. All the samples were

applied once daily for 16 days, starting from

the day of wounding. The observations of

percentage wound closure were made on 4th

,

7th

, 10th

, and 13

th, post wounding days. The

wound area of each animal was measured by

using tracing paper methods. The percentage of

wound contraction was calculated from the

days of measurements of wound area

(Shirwaikar A. et al., 2003).

Wound contraction:

The wound contraction was calculated as

percentage reduction in wound area with

respect to initial wound area while the

epithelization time was noted as the number of

days after wounding required for scar to fall off

leaving no raw wound behind.

Incision wound model:

In the incision wound model, mice

depilated by removing hairs at the dorsal

thoracic region before wounding. Mice were

anaesthetized by diethyl ether prior to incision.

Six centimeter long paravertebral incisions

were made through full thickness of skin on

either side of vertebral column of the mice. The

wounds were closed with interrupted sutures of

one centimeter apart.

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 381–388

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

The mice are categorized in to four groups

(n=6).the animals of group l treated as control

and only ointment base applied topically. The

animal of group II treated as standard drug and

third group treated as polyherbal applied

topically, respectively. All the samples were

applied once daily for 13 days, starting from

the day of wounding. The sutures were

removed on 8th

post wounding day. The tensile

strength of wounds was measured on 10th

day

following continuous water flow technique

(Shirwaikar A. et al., 2003).

Tensile strength in incision wound model:

The tensile strength was calculated in

incision wound model. On 10th

day the mice

were again anesthetized and each mice is

placed on a stack of paper towel on the middle

of the board. The amount of the towel could be

adjusted in such a way so that the board. The

amount of the towel could be adjusted in such a

way so that the wound is on the same level of

tips of the arms. The clamps are then carefully

clamped on the skin of the opposite side of the

skin of wound at a distance of 0.5 cm away

from the wound. The longer pieces of the

finishing line are placed on the pulley and

finally to the polyethylene bottle and the

position of the board is adjusted so that the

bottle receive a rapid and constant mice of

water from the large reservoir,until the wound

began to open. The amount of water in

polyethylene bag is weighted and consider as

tensile strength of the wound (Kamano et al.,

1994).

Histopathology study

On 13th

day some of animals under each

group were sacrificed and wounds were excised

together with surrounding skin. The 1µ thin

paraffin section of wounds bed material were

fixed in 10% neutral buffer formalin and

histological evaluation was performed on

heamotoxylin and eosin stain.

After complete staining the slides,

microscopic photographs of collagen tissue

were taken as were shown in figure for control,

standard and treated.

Histological studies of granulation tissue of

the methanolic extract treated animals showed

significant increase in collagen deposition with

macrophages, fibroblast, and blood vessels as

compare to control.

Statistical analysis

All the data are expressed as mean ± SD.

The values obtained for the extracts were

compared with control group using one way

ANOVA followed by tukey’s test. The values

of P≤0.001 were considered to indicate a

significant difference between the groups.

RESULTS

The methanolic extracts obtained by soxhlet

solvent extraction were subjected to various

qualitative tests to detect the presence of plant

constituents. Alkaloids, Glycosides,

Carbohydrates, Phytosterols, Saponins, and

Tannins, phenolic compounds, Proteins, free

amino acids and Flavonoids are shown in table

1. Excision wound heal by contraction process

closure and epithelization the percentage of

wound closure or closure rate include by

recording the changes in wound area at fixed

intervals of time 4th

, 7th

, 10th

and 13th

days in

fig.1 after treating with methanolic extract the

percentage of wound closure on the 13th

day

was 1.67 ± 0.573 mm2

(table 2). Incision

wounds heals by granulation and collagenation

the mean wound breaking strength or tensile

strength of wound in control group was 172.2

± 1.7049 gm while in the case of methanolic

extract treated group it was 381.22 ± 0.572 gm.

The granuloma tissue dissected out was

subjected to histopathological examination in

control group which revealed presence of

chronic inflammatory cells, edema cells. and

blood vessels were under developed. Collagen

appeared to be incomplete and improper in

growth. In methanolic extract treated group a

bulk of collagen was seen with fewer amounts

of inflammatory cells. Collagen maturity was

better than the control group blood vessels were

developed (Figure 2).

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 381–388

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

TABLE 1. Phytochemical screening of various extract of A. macrorrhizos

S.No Test Alocasia macrorrhizos

(Methanolic Extract)

1 Test for steroids (1. Salkowaski test) +

2. Test for glycosides (1.bontrager 2.kellar-killiani test

3.legal’s test

+++

3. Test for saponins (foam test) +

4. Test for carbohydrate (1.molisch’s test 2. Barfoard’s

test 3. Fehling test 4.tollen’s test

−

5. Test for alkaloids(1.mayer’s test 2.wagner test

3.dragondroff’s test 4.hagger’s test)

−

6. Test for flavonoids (ferric chloride) +

7. Test for tannins (1.ferric chloride 2. Gelatin test) ++

8. Test for protein (1.precipitation 2.xanthoproteic) +

TABLE 2. Percentage wound contraction in excision wound model

Area of wound closure (sq mm ± S.D)

Group 4th

day 7th

day 10th

day 13th

day Epithelization

period (days)

I (control) 78.5 ± 0.54

(10.75%)

69.2 ± 0.469

(15.4%)

38.2 ± 1.97

(30.9%)

26.3 ± 1.97

(36.85%)

22

II(standard) 54.6 ± 1.502

(22.7%)

28.8 ± 0.59

(35.6%)

10.1 ± 1.068

(44.95%)

0.62 ± 0.51

(49.69%)

12

III(treated) 56.04 ± 1.50

(21.98%)

39.8 ± 1.03

(30.1%)

21.9 ± 1.00

(39.05%)

1.67 ± 0.573

(49.16%)

13

p≤ 0.01 v5 control

p≤ 0.01 indicates significant when compared with control.

¥ Figure in parenthesis indicates percent wound contraction.

TABLE 3. Tensile strength in incision model

Group Tensile strength (in gram)

Control (I) 172.2 ±1.704

Standard (II) 405.32 ± 134.94

Treated (III) 381.22 ± 0.572 P≤0.05v5 control

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 381–388

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Figure 1. Photography of the wound contraction of 7 days old wound

Wound contraction of 7 days old wound in Control group (A), 7 days old wound in Standard group (B), 7 days old

wound in Treated group (C), 13 days old wound in Control group (D), 13 days old wound in Standard group (E), 13 days

old wound in Treated group (F).

Figure 2.: Histopathological photography

Histopathological photography of Standard group (A), Treated group (B), Control group (C)

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 381–388

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

DISCUSSION

Wound healing process consists of different

phases such as granulation, collagenation,

collagen maturation and scar maturation which

are concurrent but independent to each other.

Hence in the study three different models were

used to assess the effect of herbal ointment on

various phases.

The result showed that methanolic extract

possesses a definite prohealing action. This was

demonstrated by significant increase in the rate

of wound contraction and by enhanced

epithelialization. Significant increase (P<0.001)

in tensile strength and collagen levels were

observed, which was further supported by

histopathological studies and gain in granuloma

breaking strength.

The effect of methanolic extract of A.

macrorrhizos were screened on excision,

incision and histopathological wound models

concurrently with the control and reference

standard. betadine treated animals showed

more pronounced wound healing activity than

methanolic extract treated animals. The rate of

wound contraction was faster in these and

complete epithelialization of the excision

wound was observed on 13th

day. In standard

betadine treated animals complete

epithelialization was noticed on 12th

day. The

result of percentage wound contraction and

period of complete epithelialization has been

depicted in table 2. In incision model

significant increase in tensile strength of healed

wounds was observed in methanolic extract

treated group (381.22 ± 0.572). Histological

studies of granulation tissue on control group of

animals showed accumulation of more

macrophages and very few collagen fibers. In

methanolic extract treated group, histological

section of granulation tissue showed very few

macrophages and showed complete

epithelialization and collagen where in standard

group showed complete collagen.

CONCLUSION

In conclusion, the significant increase in

tensile strength and the prominent haemostatic

activity exhibited by the methanolic extract of

A. macrorrhizos shows the potent wound

healing property of plant. Further more detailed

investigation on the actives responsible for the

activity are to be identified and further more

clinical researches should be conducted to

bring the effects of the drug out to the scientific

world.

REFERENCES

Fabricant D.S., Farnsworth N.R. (2001). The

value of plants used in traditional

medicine for drugdiscovery, Environ

Health Pers, 109 (Suppl 1), p. 69–75.

Krishnan P. (2006). The scientific study of

herbal wound healing therapies: Current

state of play. Curr.Anaesthesia Crit.

Care.17, p.21–27.

Lazarus G.S. Cooper D.M. KInghton D.R.

Margolis D.J. Pecoraro R.E.

Rodeheaver G. Robson M.C. (1998).

Defination and guidelines for

assessment of wounds and evaluation of

healing.Arch. Dermatol.; 130, p. 49–

493.

Li J. Chen J. Kirsener R. (2007).

Pathophysiology of acute wound

healing.Clin. Dermatol. 25, p. 9–18.

Madden J.W. Peacock E.E. (1968). Studies on

the biology of collagen during wound

healing. I. Rate of collagen synthesis

and deposition in cutaneous wounds of

the rat. Surgery, 64, p. 288–294.

Menke N.B. Ward K.R. Witten T.M. Bonchev

D.G. Diegelmann R.F. (2007). Impaired

wound healing Clin.Dermatol. 25, p.

19–25.

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 10 | October 2014 | 381–388

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Perumal S.R., Ignacimuthu S., Patric R.D.

(2008). Preliminary screening of

ethnomedicinal plants from India. Eur

Rev Med Pharmacol Sci.12, p. 1–7.

Principe P. (2005).Monetising the

pharmacological benefits of plants. US

Environmental protection

Agency,Washington, D.C. 1991.

Priya K.S. Gnanamani A., Radhakrishnan N.,

Babu M. (2002). Healing potential of

Datura Alba on burn wounds in albino

rats. J. Ethnopharmacol. 83, p.193–199.

Prockop D.J. Kivirikko K.I. Tuderman L.

Guzman N.A. (1979). The biosynthesis

of collagen and its disorders.N.Engl. J.

Med. 301, p. 13–23.

Purna S.K. Babu M. (2000). Collagen based

dressings/a review. Burns. 26, p. 54–62.

Ramzi S.C., Vinay K., Stanley R.

(1994).Pathologic Basis of Diseases.5th

edition, WB Saunders Company,

Philadelphia, p. 86.

Schultz G.S. (1999). Molecular Regulation of

Wound Healing In: Acute and Chronic

Wounds: Nursing management. Bryant

R.A., 2nd Edition, WB Saunders

Publisher, USA, p.413–429.

Shirwaikar A., Shenoy R., Udupa A.L., Udupa

S.L., Shetty S. (2003). Activity of

wound healing. Indian journal of

Experimental Bilogy, 41, p. 238−241.

Stadelmalmann W.K. Digenis A.G. Tobin G.R.

(1998).Physiology and healing

dynamics of chronic cutaneous

wounds.Am. J. Surg. 176, p. 26S–38S.

Steenkamp V., Mathivha E., Gouws M.C.,

Rensburg C.E.J. (2004). Studies on

antibacterial, antioxidant and fibroblast

growth stimulation of wound healing

remedies from South AfricaJ.

Ethnopharmacol.95, p.353–357.

Strodtbeck F. 9 (2001).Physiology of wound

healing.Newborn Infant Nurs. Rev. 1, p.

43–45.

Source of Support: NIL Conflict of Interest: None Declared

Call for Papers – Vol. 3, Issue 12, December 2014

Submit your manuscripts (Research articles, Review

articles, Short Communications, Letters to the Editor,

Book Reviews) to Global Journal of Research on

Medicinal plants & Indigenous medicine – GJRMI

Submit it online through www.gjrmi.com or mail it to

submitarticle@gjrmi.com on or before

November 10th

2014.

To advertise on the Flip book Cover page freely,

write to

chiefeditor@gjrmi.com or editorinchief@gjrmi.com

Or

Call - +919590574495