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INDEX – GJRMI, Vol.2, Iss. 6, June 2013

MEDICINAL PLANTS RESEARCH

Botany SUPPRESSION OF LEAF BLIGHT PATHOGEN ALTERNARIA LONGIPES OF MEDICAGO

SATIVA BY FLUORESCENT PSEUDOMONAS STRAIN BRL-1

Sen Surjit, Acharya Krishnendu 401–409

Animal Production Technology SENSITIVITY OF BUCK (MALE GOAT) SEMEN MICROBIAL ISOLATES TO THREE PLANT

EXTRACTS FROM CAMEROON

Salah A Martin, Yongabi K A 410–417

Review Article

A BRIEF REVIEW ON GINKGO BILOBA L. (MAIDENHAIR TREE) - A RARE MULTIPURPOSE

MEDICINAL PLANT

Patel Hemangi, Ingalhalli Rajashekhar 418–427

INDIGENOUS MEDICINE

Ayurveda – Kaumarabrithya EFFECT OF BADARA STEM BARK (ZIZIPHUS JUJUBA LAMB.) IN THE MANAGEMENT OF

VATAJA KASA IN CHILDREN

Chethan Kumar V K, Shailaja U

428–434

Ayurveda – Dravya Guna

EFFECT OF PARNAYAVANI (COLEUS AMBOINICUS LOUR.) ON MES INDUCED EPILEPSY IN

RATS

Sharma Monica, Khemani. N, Singh J 435–440

Ayurveda – Dravya Guna

ANTIMICROBIAL POTENTIAL OF THE WILD AND CULTIVATED VARIETY OF ERANDA

(RICINUS COMMUNIS LINN.) ROOT

Doshi Krunal A, Sagar Dhwani H, Acharya R N 441–447

Ayurveda – Review Article – Kaumarabhritya

PROBABLE ETIOPATHOGENESIS (SAMPRAPTI) OF AUTISM IN FRAME OF AYURVEDA IN

RELATION TO INTENSE WORLD THEORY

Yadav Deepmala, Behera Banshidhar, Kumar Abhimanyu 448–459

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DOMAIN

Vaikos C D, Waghchoure Ashok 460–464

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TRADITIONAL USE OF KAHU (LACTUCA SCARIOLA L.) - A REVIEW

Arif Mohammad 465–474

HIMALAYA HERBAL HEALTH CARE’S quarterly INFOLINE endorses & features Global Journal of

Research on Medicinal plants & Indigenous medicine (GJRMI) in their recent Vol. 9, Jan-mar 2013 issue.

The Editor-in-Chief, GJRMI, on behalf of the Board members & Referees, thank the scientific publications

division, The Himalaya Drug Company for the endorsement.

Click this Face book Link to read the below material

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 401–409

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

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

SUPPRESSION OF LEAF BLIGHT PATHOGEN ALTERNARIA LONGIPES

OF MEDICAGO SATIVA BY FLUORESCENT PSEUDOMONAS

STRAIN BRL-1

Sen Surjit1, Acharya Krishnendu

2*

1, 2Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata 700 019, INDIA

*Corresponding Author: [email protected]; Fax : +91 033 24764419; Phone : +91 8013167310

Received: 05/04/2013; Revised: 17/05/2013; Accepted: 26/05/2013

ABSTRACT

A leaf blight disease caused by the fungus Alternaria longipes was observed in Medicago sativa

Linn. commonly known as ‘Alfalfa’, a tonic plant cultivated at different areas of West Bengal, India.

A potential biocontrol agent viz. fluorescent Pseudomonas strain BRL-1 showed both in vitro and in

vivo antagonistic activity against the pathogen. In dual culture bioassay as circular and semicircular

patterns, the isolate quantitatively inhibits the growth of the pathogen by about 74% and 65%,

respectively. Foliar application of a talc-based formulation of the antagonist to field condition

revealed that the maximum mean disease index reached to 1.097 and 1.19 in 2010 and 2011,

indicating around 77% and 76% reduction in disease severity when compared to non-treated control.

Transferring the outcome of this work in the field would benefit the growers by facilitating

bioorganic production of this important medicinal plant and finally consumers can get hazard free

natural plant product.

KEYWORDS: Alfalfa, biocontrol, leaf blight, medicinal plant

Research article

Cite this article:

Sen. S., Acharya K., (2013), SUPPRESSION OF LEAF BLIGHT PATHOGEN ALTERNARIA

LONGIPES OF MEDICAGO SATIVA BY FLUORESCENT PSEUDOMONAS

STRAIN BRL-1, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 401–409

mailto:[email protected]



INTRODUCTION

Medicago sativa (Alfalfa) is a perennial

flowering plant in the family Fabaceae

cultivated as an important forage as well as

medicinal plant in many countries. It is used as

tonic due to presence of high percentage of

proteins (60.5%), minerals, enzymes, vitamins

etc. Alfalfa is a valuable source of vitamins A

and E, fresh is rich in Vitamin C (1.78 mg/g)

but it loses 80% of vitamins on drying (Rashmi

et al., 1997). The leaves are also rich in vitamin

K, which is used medicinally to encourage the

clotting of blood (Weiner, 1980). Alfalfa

leaves, either fresh or dried, have traditionally

been used as a nutritive tonic to stimulate the

appetite and promote weight gain (Foster and

Duke, 1990). The plant is antiscorbutic,

diuretic, oxytocic, haemostatic, nutritive,

stimulant (Duke and Ayensu, 1985). The

expressed juice is emetic and is anodyne in the

treatment of gravel (Duke and Ayensu, 1985).

The plant is taken internally for debility in

convalescence, anaemia, haemorrhage,

menopausal complaints, pre-menstrual tension,

fibroids etc (Bown, 1995; Chevallier, 1996). A

poultice of the heated leaves has been applied

to the ear in the treatment of earache

(Moerman, 1998). The plant is grown

commercially as a source of chlorophyll and

carotene, both of which have proven health

benefits (Foster and Duke, 1990). The leaves

also contain the anti-oxidant tricin (Foster and

Duke, 1990). The root is febrifuge and is also

prescribed in cases of highly coloured urine

(Duke and Ayensu, 1985). Extracts of the plant

has antibacterial activity (Duke and Ayensu,

1985).

Paul and Singh, (2002) reported fungal

phytopathogens attack medicinal and aromatic

plants leading to significant quantitative and

qualitative loss. Alternaria leaf blights were

very common in medicinal plants cultivated in

various districts of West Bengal, India (Maiti et

al., 2007a, b, c). Today biocontrol agents are

gaining importance in the field of disease

management of medicinal plants as they do not

have the adverse effects like that of fungicides

which eventually contaminate the purity of

plant drugs (Mathivanan et al., 2005).

Extensive research is underway globally to

exploit the potentiality of pseudomonads,

which help to protect crops from

phytopathogens and are metabolically and

functionally more diverse (Choudhury et al.,

2009). A wide range of fluorescent

pseudomonads have been reported for having

in vitro and in vivo biocontrol potentiality

against variety of phytopathogens (Kishore et

al,. 2005; Mansoor et al., 2007; Sen et al.,

2009; 2012; Maiti et al., 2012). The present

study demonstrates the efficiency of a potential

biocontrol agent fluorescent Pseudomonas

BRL-1 for controlling leaf blight disease in

Medicago sativa.

MATERIALS AND METHODS

Organisms

The pathogenic organism was isolated from

the diseased leaves of M. sativa as a pure

culture on potato dextrose agar medium (PDA),

identified as Alternaria longipes (Maiti et al.,

2007b). The culture was maintained in the

same medium and stored at 4°C for further

study. The biocontrol agent fluorescent

Pseudomonas BRL-1 was obtained from our

laboratory culture stock. The antagonist was

subcultured and maintained on tryptic soy agar

(TSA) medium for subsequent use.

Dual Culture Bioassay

Fluorescent Pseudomonas BRL-1 from 24 h

old culture (107 cells ml

-1) was streaked in the

peptone (1%) glucose (2%) agar (2%) (PGA)

plate as circular / O and semicircular / U

pattern. Then mycelial disc (5 mm diameter) of

3 days old culture of A. longipes was

subsequently inoculated at the center of O or U

shaped region on the PGA plates (Skidmore

and Dickinson, 1976). Inoculation only with

the pathogen served as control. The plates in

triplicate were incubated at 30°C for 5 days and

diameter of colony growth was measured at

every 24 h intervals. Light microscopic (Zeiss

AX 10) studies were also performed to detect

physical and / or morphological changes of

mycelia.



Talc based formulation and survival of

fluorescent Pseudomonas BRL-1

Talc-based formulation of the antagonist

was prepared using a method developed by

Vidyasekaran and Muthamilan (1995)

modified. Ten gram carboxy methyl cellulose

(CMC) per kg of sterile talc was used as

adhesive. The bacterial suspension (8 × 109

CFU ml-1

) was mixed with sterile talc (400 ml

kg-1

) and air dried (approximately to 35% w/w,

moisture content). The formulation was stored

at 4°C for up to 180 days. The antagonist in the

talc-based formulation was monitored in vitro

with respect to its shelf-life and viability under

this storage condition. Survival of the bacterial

population in the formulation was assayed at 30

days intervals using King’s medium B in a

dilution plate assay according to Vidyasekaran

and Muthamilan, (1995).

Survival of fluorescent Pseudomonas BRL-1

on phylloplane

Survival and multiplication of fluorescent

Pseudomonas BRL-1 on the phylloplane of

Medicago was determined following the

method of Kishore et al. (2005). The talc-based

formulation (4 g l-1

) of the antagonist was

applied as foliar sprays with an initial cell

density of 106 CFU g

-1 of fresh leaves. Leaves

were collected at 3 days intervals, washed in

sterile water, serially diluted and plated (in

triplicate) on King’s B agar medium. Colonies

of BRL-1 was determined after 48 h of

incubation at 30°C utilizing its colony

morphology, fluorescence characteristics. The

bacterial populations were expressed as log

CFU g-1

of leaf. The whole experiment was

performed thrice.

Field studies

The Medicinal Plant Garden, R.K. Mission

Ashrama, Narendrapur, India was used for the

field experiments during 2010 and 2011 when

the environmental conditions were conducive

(January to July) for the rapid spread of A.

logipes on M. sativa. The trial was conducted

as a randomized complete block design with

three replicate plots (3 × 4 m2) and forty plants

per plot. Well-rotted farmyard manure was

mixed well into the soil before planting the

seedlings. Four-week-old disease free seedlings

were transplanted to the random blocks on end

of January allowing A. longipes leaf blight to

develop naturally (Silva et al., 2004). The talc

based formulation the antagonist was prepared

by dissolving it in water (4 g l-1

) allowing it to

settle for 1 h, and filtering the solution through

muslin cloth. The filtrate was applied as a foliar

spray using a low volume sprayer beginning at

transplant and repeating every 15 days for up to

six months i.e. end of July 2010 and 2011.

Plots sprayed with the talc-based carrier

without the biocontrol agent was served as

control. Twenty-five plants from each plot were

rated for disease severity at 15 day intervals

starting at transplant for up-to six months using

a 0-5 rating scale (Kishore et al., 2005).

Statistical analysis

The disease severity data were statistical

analysed by using analysis of variance

(ANOVA) followed by Tukey’s Test to find

out significance level at 1% (p<0.01).

RESULTS

Dual culture of fluorescent Pseudomonas

BRL-1 and A. longipes

Growth inhibition of A. longipes by strain

BRL-1 was observed in dual culture. Mycelial

growth was restricted near bacterial streak and

continued away from it. The growth inhibition

of A. longipes remained proportionate with an

increased incubation period of up to 5 days.

Quantitatively fluorescent Pseudomonas BRL-

1 inhibited the growth of A. longipes by

73.76% and 64.66% in circular and

semicircular streaks after 120 h of incubation

respectively (Fig. 1). After 120 h of

confrontation, microscopic examination of the

mycelia at the inhibition zone showed

markedly damaged hyphae as evidenced by

disorganization of the cytoplasm, shriveling,

growth deformities, swelling, fragmentation,

short branching and lysis (Fig. 2).



Fig. 1 Inhibition of A. longipes by fluorescent Pseudomonas BRL-1 under dual plate culture

using circular (O) and semicircular (U) method. Each point represents the mean ± SE

(standard error) of three separate experiments, each in triplicate.

0

10

20

30

40

50

60

70

80

0h 24h 48h 72h 96h 120h

Incubation, h

% I

nh

ibit

ion

of

rad

ial

my

ce

lia

l g

row

th

O Shaped

U Shaped

Fig. 2 [A] Microscopic observation of mycelium of control plate showing normal hyphal

structure and spore [B] Microscopic observation of mycelium of treated plate showing hyphal

deformities, short branching, and disorganization of hyphal cytoplasmic contents.



Fig. 3 Survival of fluorescent Pseudomonas BRL-1 in talc based formulations stored at 4°C.

Each point represents the mean ± SE (standard error) of three separate experiments, each in

triplicate.

5

5.5

6

6.5

7

7.5

8

8.5

9

0 30 60 90 120 150 180

Storage at 40C

log

cfu

/g o

f ta

lc b

ased

fo

rmu

lati

on

Fig. 4 Changes in the populations of fluorescent Pseudomonas BRL-1 on M. Sativa phylloplane

growing under field condition. Each point represents the mean ± SE (standard error) of three

separate experiments, each in triplicate.

2

3

4

5

6

7

0 5 10 15

Days after inoculation

log

CF

U / g



Table 1 Field study for the control of leaf blight disease of M. sativa caused by A. longipes in

2010 and 2011. Foliar application of the talc based formulation on date of transplantation and

at an interval of 15 days until 180 days. Disease index was rated on a 0–5 scale. Values are

mean ± SE of twenty-five randomly selected plants per plot of three individual plot

experiments

Days after

transplantation

Disease index

2010

Disease index

2011

Control Treated Control Treated

0 0 0 0 0

15 0.159 ± 0.032 0.102 ± 0.016* 0.182 ± 0.009 0.162 ± 0.006*

30 0.254 ± 0.014 0.183 ± 0.007* 0.358 ± 0.024 0.213 ± 0.009*

45 0.678 ± 0.021 0.321 ± 0.012* 0.773 ± 0.014 0.424 ± 0.016*

60 0.947 ± 0.006 0.578 ± 0.014* 1.047 ± 0.019 0.612 ± 0.015*

75 1.542 ± 0.019 0.634 ± 0.012* 1.96 ± 0.017 0.69 ± 0.033*

90 2.126 ± 0.015 0.719 ± 0.012* 2.724 ± 0.044 0.794 ± 0.017*

105 3.241 ± 0.014 0.801 ± 0.018* 3.81 ± 0.017 0.912 ± 0.009*

120 3.734 ± 0.008 0.886 ± 0.005* 3.965 ± 0.026 0.98 ± 0.014*

135 4.225 ± 0.033 0.915 ± 0.018* 4.45 ± 0.014 1.03 ± 0.006*

150 4.412±0.011 0.988±0.024* 4.561±0.032 1.087 ± 0.024*

165 4.61 ± 0.009 1.029 ± 0.015* 4.73 ± 0.019 1.12 ± 0.016*

180 4.76 ± 0.012 1.097 ± 0.016* 4.88 ± 0.014 1.19 ± 0.015*

* Data in each row differ significantly with control according to Tukey’s test (P<0.01)

Survival of fluorescent Pseudomonas BRL-1

in talc-based formulation and on the M.

sativa phylloplane

Survival of the antagonist in talc-based

formulation stored at 4°C was monitored for

180 days (Fig. 3). With time, the initial

population of BRL-1 (8.7 log CFU g-1

) in talc

based formulation gradually decreased.

Approximately 30% decrease in the colony-

forming unit was estimated on 180th

day after

storage at 4°C. Foliar application of this

formulated antagonist showed the initial

population (within 1 h after application) to be

log 6.4 CFU g-1

of leaves. The initial

population steadily declined to log 3.3 CFU g-1

of leaves at 15th

day after treatment (Fig. 4). No

colonies of BRL-1 were recovered on the

controls.

Field studies

Talc based formulation of fluorescent

Pseudomonas BRL-1 was evaluated in the field

on M. sativa for two successive seasons. After

the 3rd

and 4th

spray of the formulation, new

symptoms of leaf blights were inhibited in

treated plots. The mean disease index after 180

days in control field reached to 4.76 and 4.88 in

2010 and 2011, respectively, where most of all

plants were severely affected and more than

60% leaves were defoliated (Table 1). On the



contrary, the disease index in BRL-1 treated

fields reached only 1.097 and 1.19 in 2010 and

2011, indicating around 77% and 76%

reduction in disease severity respectively.

DISCUSSION

Over the past twenty years, the control of

plant pathogenic fungi by antagonistic bacteria

and fungi has been the topic of numerous

studies. The majority of these studies dealt with

antagonists controlling soil borne pathogens

and to a less significant extent, foliar

pathogens. Pseudomonas strains were

evaluated for their ability to control Sclerotinia

homeocarpa and Bipolaris sorokiniana on the

phylloplane of Kentucky bluegrass (Hodges et

al., 1994). Pseudsomonas, Bacillus and

Stenotrophomonas showed in vitro antifungal

activity against Verticillium dahliae var.

longisporum (Berg et al., 1998). Rozsnyay et

al. (1992) showed that some strains of P.

fluorescens and some fungi inhibited canker

and dieback diseases of apricot. The strain

BRL-1 have been reported as a potential

biocontrol agent for controlling both soil borne

and aerial pathogen as evidenced through in

vivo and field experiments conducted earlier

(Sen et al., 2009; 2012).

An effort to characterize the mycoparasitic

action of BRL-1 in the present study revealed

that the bioagent had the potentiality to manage

A. longipes both in vitro and in vivo. Recently it

has been reported that the same antagonist had

the ability to control leaf spot disease of Stevia

rebaudiana, caused by Alternaria alternata

(Sen et al., 2012). Coagulation of cytoplasmic

contents and disorganization of fungal hyphae

upon dual-culture attributed by the strain BRL-

1 mainly by the release of secondary

metabolites like siderophore, Indole Acetic

Acid (IAA) and lytic enzymes (Sen et al.,

2012). Similar type of observations were made

earlier by Bano and Musarrat (2002), Kapsalis

et al. (2008), Ramyasmruthi et al. (2012) Maiti

et al. (2012) who demonstrated that the

exposure of different phytopathogenic fungi to

secondary metabolites and lytic enzymes such

as chitinase and protease could result in the

degradation of the fungal cell wall. IAA reduce

spore germination, mycelial dry weight and

protein content of the pathogenic fungi and thus

prevent significantly any chance for disease

induction (Sharaf and Farrag, 2004; Sen et al.,

2009). According to several studies,

siderophore, chitinase and protease produced

by fluorescent pseudomonads are known to

inhibit the growth of some fungal pathogens

(Lim and Kim 1995; Kapsalis et al. 2008; Maiti

et al. 2012).

CONCLUSION

In the present study, we showed that the

strain BRL-1 could live well in talc-based

formulation stored at 4°C for 6 months. It has

been reported that fluorescent pseudomonads

could stay alive in certain formulations

(Connick 1988; Mugilan et al. 2011).

Moreover, the success of any biocontrol

formulation not only depends on the ability of

the biocontrol agent to survive in the

formulation, but also its capacity to survive on

the host plant to which it is applied. Bunster et

al. (1989) and Lindow (1995) reported foliar

application of fluorescent pseudomonads are

able to survive well on leaf surfaces by

occupying particular surface sites. Our study

complements their findings by demonstrating

that an average of 53% cell population survived

for 15 days after application of talc-based

formulation of BRL-1 on M. sativa. The

formulation when applied in every 15 days, for

two consecutive seasons successfully reduced

symptoms of A. longipes leaf blight on M.

sativa by 77% and 76% respectively. This

ultimately helps the farmers to limit the use of

hazardous fungicides and simultaneously

saving their crops. Finally, the concerted effort

of academic, federal and private sector

scientists should lead to the development of

effective and consistent biocontrol of aerial

plant diseases based on an integrated biological

control strategy.



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Bano N, Musarrat J (2002). Characterization of

a new Pseudomonas aeruginosa NJ-15

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Source of Support: Nil Conflict of Interest: None Declared




SENSITIVITY OF BUCK (MALE GOAT) SEMEN MICROBIAL ISOLATES

TO THREE PLANT EXTRACTS FROM CAMEROON

Salah A Martin1*, Yongabi K A

2

1 Department of Animal Production Technology, College of Technology, University of Bamenda, Cameroon,

P.O Box 39 Bamenda 2 Phyto-biotechnology Research Foundation, P.O. Box 9221, Bamenda-Cameroon

*Corresponding Author: E-mail: [email protected]


ABSTRACT

A study was conducted to investigate the prevalent microorganisms in Buck semen in Bouchi.

Ten semen collections from three Bucks (BI, BII, BIII) of the Red Sokoto Breed were cultured

aerobically on nutrient agar and MacConkey agar (oxoid), and blood agar (oxoid) for bacteria on

potato destrose agar (oxoid) for fungi. Two bacterial species Staphylococcus aureus and Bacillus

cereus together with mould Aspergillus niger were used. Aspergillus fumingatus and Torulopsis spp

were isolated. The corresponding pH values and nitric contents were indicative of abnormality and

infection respectively. The isolates from the samples were tested against three different herbs

(Carica papaya L., Vernonia amygdalina Delile., and Jatropha curcas L. A cold ethyl acetate extract

of the leaf and stem bark of the herbs showed appreciable inhibition on all the isolates except

Jatropha curcas leaf that never showed an effect

KEY WORDS: Artificial insemination, semen, buck herbs, Carica papaya L., Vernonia amygdalina

Delile., and Jatropha curcas L. and Antimicrobials

Research article

Cite this article:

Salah. A. Martin, Yongabi. K. A. (2013), SENSITIVITY OF BUCK (MALE GOAT) SEMEN

MICROBIAL ISOLATES TO THREE PLANT EXTRACTS FROM CAMEROON,

Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 410–417



INTRODUCTION

Artificial insemination is a very important

practice by animal scientists, and remains true

in Nigeria. During this practice, semen

collected is normally preserved in specially

compounded diluents pending insemination in

the practice of artificial insemination in the

country. Animal scientist often includes

antibiotics and sometimes antimycotics in the

diluents before insemination.

This is done to protect the destruction of the

semen content before insemination, but

unfortunately; the inclusion of antibiotics has

often been carried out without knowledge of

the kind of bacteria present in the semen during

ejaculation (Jimenez et al., 2012). Above all,

some insemination do not bother to include

antimycotics even then their pathogenic role in

the semen with regard to the release of toxin

(aflatoxin and mycotoxins is crucial to the

reduction of the semen viability. Besides, some

bacteria in the semen may develop resistance

due to genetic adaptations (Colleuau et al.,

2011, Nordstoga et al., 2011) to some of the

antibiotics normally used in diluents.

Studies in Cameroonian medicinal plants

research is a very active domain in our

Universities (Salah et al., 2012, Ndip et al.,

2007) especially in the domain if antimicrobial

studies ( Asob et al., 2011, Noumedem et al.,

2013). The occurrence of microorganisms and

their pathogenic role in certain cases have been

exclusively reviewed (Akiynanju et al., 1986)

by blood and radotitls and Zgnorniak

Nowosienka et al., 1984). Pathogens and other

microflora have been shown to have adverse

effects on the fertility of semen by the

production of toxins, putrefaction of diluents,

components of biological origin, and the

utilization of metabolic substrate, lamming

(Tuncer et al., 2013). In the practice of artificial

insemination, the possible pathogenic effects

within the female reproductive tract as well as

semen viability during storage of these

microbes have been controlled considerably by

the use of antibiotics and antimycotics in

diluents (Butswat and Choji, 1974). However,

certain constraints have militated against the

adoption of artificial insemination in this zone

(Audu, 1989) and the farmers have had to

depend on natural mating which exposes the

female to contamination from males carrying

pathogenic organisms within their urino-genital

tract. The scarcity of veterinary services, the

high cost of drugs particularly antibiotics and

antimycotics for inclusion in the diluents as

well as for treating animals, and the lack of

knowledge for the microbial spectrum of semen

are some of the factors that have the space of

the adoption of the option of artificial

insemination.

This study investigates the prevalent

microorganisms in Buck semen of the Red

Sokoto breed and exploits indigenous means of

controlling them with medicinal herbs.

MATERIAL AND METHODS

Semen Collection

Semen was collected from July to

November 2006 using the artificial vagina

(AV) method after thorough feeding and

abstinence from three months. The AV was

normally washed with soap and hot water after

each collection and disinfected with 70%

alcohol after drying (Butswat and Choji, 1994)

Isolation and Identification of

Microorganisms

Bacteria

A loopful of the test semen was aseptically

streaked on nutrient agar MacConkey agar and

blood agar plates and incubated aerobically at

37oC for 24 h. This was repeated for ten

collections with the pH and nitrite values

determined, using diagnostic urinalysis test trip

(Combi-9 test strip) according to standard

medical diagnostic method. Bacterial colonies

were carefully picked and purified by repeated

sub-cultures on nutrient agar and blood agar

plates and their morphologies were studied.

Pure cultures were preserved on nutrient

agar slants and used for gram staining and other



biochemical test (Chesborough, 2008), and the

isolates were identified according to methods

of Buchana and Gibbons (1974) and Harringan

and McCance (1976)

Fungi

A loopful of the test semen was aseptically

inoculated onto potato dextrose agar plates

(Oxoid Ldt) in triplicates and incubated at 35oC

for 5 days. This was repeated for ten collections

for the three bucks.

Following incubation, the growth were

examined using an hand lens (10x) before

picking out specimens in lactophenol cotton

blue on a microscope slide and examined

according to the methods of Harrigance and

McCance (1976) and latter identified according

to methods of Barnett and Hunter (1972).

Preliminary Screening of herbs with

antimicrobial Property on Isolates

Samples Collection

Herbs were collected from villages around

Bouchi; Yelwa Tudu, Kagadamia, Doka and

Dumba in Gwallameji. Those described by

Audu were adopted from for the selection of

plants (2). Leaves were used and samples were

deposited in the herbarium of the institution.

Processing of Plant Materials

Plant parts were dried under shade for two

weeks and then carefully packed in large Khaki

enveloped and dried in an oven at 37oC for 12 h

before milling. Fine powders were obtained

through sieving. Crude extracts of the plant

powder were made by adopting a modified

existing method of Akinyanju et al., 1986 and

Audu 1987. Cold solvent extraction was used.

Ten grams each of the fine powder were

weighed and soaked in 50 ml of ethyl acetate in

the ratio 1:5 weight/volume and allowed to

stand and for 48 hours at 25oC. Gravity

filtration was done using filter paper (Whatman

No. 13) and evaporated to dryness in the sun.

Antimicrobial Screening Test on Isolates

The bacterial isolates were cultured in

peptone water for 18 hrs. 0.3 ml of each of the

bacterial suspension was mixed with 15 ml of

nutrient agar in sterile petri-plates and allowed

to solidify. A sterile steel borer was used to

punch wells into the agar and each well was

filled with 0.1 ml extract (at 2% concentration)

and with 0.1 ml of distilled water as control.

This was incubated at 37°C for 24 hours and

the diameter of the zone of inhibition in

measured to the nearest mm using vernier

caliper according to methods of Akinyanju et

al., (1986).

Spores of the isolated fungi were harvested

from stock and seeded onto dextrose agar plates

with 0%, 2%, 4% and 16% concentrations of

the herbal extracts. Spore germination and

growth rate were monitored for a week.

RESULTS

Bacteria isolated from the semen samples

for the three bucks were Staphylococcus aureus

and Bacillus cereus, while Aspergillus niger

(Var tieghem, 1867), Aspergillus fumigates

(Fresenius, 1863), and Toruopsis spp were

moulds of yeast respectively (Tables I and II).

All the semen from the bucks were acidic and

buck I revealed higher acidity (pH = 5.1)

The nitrite values for the three bucks were

high; with buck I revealed a higher value

(Table III).

The diameter of zone of inhibition of

ethylacetate extract of herbs on bacteria isolate

from buck semen is shown in Table IV.

The susceptibility of the bacterial isolates to

the ethyl acetate extract of the herbs (Carica

papaya, Vernonia amygdalina, and Japtropha

curcas) and the antimicrobial activity of the

combined ethyl acetate extract of the individual

herbs on both bacterial and fungal isolates is

shown on Table V.



TABLE I: Bacterial Isolates from the three buck seminal fluid in triplicate

REPLICATE I II III

BUCK I S. a. , B.c. S. a. , B.c. S. a. , B.c.

BUCK II S. a., S. a. S. a.

BUCK III S. a., B.c S. a. , B.c S. a., B.c

S. a. – Straphylococcus aureus

B.c. – Bacillus cereus.

TABLE II: Mould and Yeast isolates from the three-Buck seminal fluid in triplicate.

REPLICATE I II III

BUCK I T. s. T. s. T. s.

BUCK II A. f A. f A. f

BUCK III A. n. A. n. A. n.

T. s. = Torulopsis spp

A. f. = Aspergillus fumingatus

A. n. = Aspergillus niger.

TABLE III: The Biochemical content of the buck semen

REPLICATE BUCK I BUCK II BUCK III

Number of samples collected 10 10 10

pH 5.1 6.1 6.1

Nitrite +++ ++ ++

N/B +++ = Highly positive

++ = Positive



TABLE IV: The diameter of zone of inhibition of ethyl acetate extract of herbs on bacteria

isolate from buck semen

Average diameter of Zone of Inhibition 9 mm for 3 replicates at 2% concentration in mm

Herbs Staphylococcus Bacillus cereus

Carica papaya

Paw-paw (Leaf)

1.0

1.0

Carica papaya

Paw-paw (stem bark)

3.0

3.0

Vernonia amygdalina

Bitter leaf (stem bark)

3.0

3.0

Vernonia amygdalina

Bitter leaf (leaf)

3.0

4.5

Jatropha curcas

Physic nut (stem bark)

4.0

3.0

Jatropha curcas

Physic nut (leaf)

Mixture of all extracts 1:1

Resistant

3.5

Resistant

6.5

Table V: The combined Effects of Ethyl acetate Plant Extracts on both Bacterial and Fungal

Isolates

Concentration (%) 2% 4% 16%

Aspergillus niger No growth from 1st –

7th

day

No growth from 1st –

7th

day


7th

day

Aspergillus

fumingatus


7th

day


7th

day


7th

day

Control Profuse growth with

sporulation

Profuse growth with

sporulation

Profuse growth with

sporulation

Discussion

The presence of nitrite in the semen

samples and above all the higher nitrite nature

of the studied semen (pH = 5.1 and 6.1) not

only confirms an abnormality, but the degree of

abnormality in the semen. It has been reported

that pH values of Friesian bull 7.0 is needed for

excellent fertility (Jimenez et al., 2012).

The pH values of the semen (pH = 5.1 and

6.1) tally with the observation of Arthur an

Allen (1972) that fresh semen samples are

slightly acidic and that excessively high values

occur with inflammation of the accessory

glands.

In a study on bacterial contamination of

boar semen, (Dede, 1981), Staphylococci were

isolated along with ten other genera of bacteria

in the tropical environment. In this present



study, Bacillus cereus, Staphylococcus aureus

as bacterial isolates while Aspergillus niger

(Vartieghem, 1867), Aspergillus fumingatus

(Fresenius, 1863) and yeast, a Toralopsis spp as

fungal isolates. This study, however, slightly

differs from the result of Zgnorniak-

Nowosielska (1984) who isolated only

saprophytic flora from horse semen sample in

Poland.

Before restraining the buck for the research

period, the animal had been feeding wildly and

perhaps might have gotten Bacillus cereus

through this means or even through the feed

used during the period of this study, as there

were no quality controls on the feed. The

mould, Aspergillus spp, might also have been

gotten through this means or may have been in

the animals for some time.

Bacillus cereus is widely distributed in

nature and lives as saprophytes in the soil, dust,

water, and on vegetation and cereals as

reported by Cheesborough (2008). It is a major

pathogen in cattle, sheep and goat and the

bacilli are excreted in the faeces, urine and

saliva of infected animals as reported by

Cheesborough (2008).

Aspergillus niger (Vartieghem, 1867) and

Aspergillus fumigates (Fresenius, 1863) are

ubiquitous saprophytes in soil, on plants, men

and animals, while the yeast, Torulopsis spp,

appear on foodstuffs and genitals of all the

animals (Noumdem et al., 2013). Fungi have

been noted to exert more effects on the

hormonal activity within the reproductive

system of animals. For instance, Aspergillus

genus was known to be associated with the

secretion of a toxic fungal metabolite,

aflatoxin. Moreso ingestion of aflatoxin-

contaminated feed could lead to widespread

reproductive abnormality in male chicken,

including a reduction in circulating levels of

testosterone (Clarke and Ottinger, 1987).

From antimicrobial screening test, papaw

leaf and stem bark (Carica papaya, Bitter leaf

and stem bark (Vernonia amygdalina) and

Jatropha curcas stem bark showed appreciable

inhibitory property against the bacteria isolates

in vitro as against no inhibition on the water

control.

Similarly, the extract of herbs combined in

equal ratios showed a much more appreciable

diameter of zone of inhibition than the

individual extracts for the bacterial isolates and

completely inhibited the growth of the fungal

spores at all test concentrations except at 0%

concentration for the first three days.

This study shows that the knowledge of

micro organism before insemination and their

sensitivities to antibiotics and antimycotics in

compounding diluents is very vital and above

all proves the possibility of indigenous

knowledge of herbs in controlling these

pathogens, so as to ease and enhance artificial

insemination practice in this environment.

However, more studies should be carried

out to map out the specific bioactive

ingredients and ways of including the herbs

directly in animal feeds or otherwise.

ACKNOWLEDGEMENT

The Authors are grateful to Dr. M. O. Agho,

Director, FEPA/ZER Centre (A. T. B. U.

Bouchi), for his assistance.

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A BRIEF REVIEW ON GINKGO BILOBA L. (MAIDENHAIR TREE)

A RARE MULTIPURPOSE MEDICINAL PLANT

Patel Hemangi1, Ingalhalli Rajashekhar

2*

1C.G Bhakta Institute of Biotechnology, Uka Tarsadia University, Mahuva Road, Bardoli-394601, Surat

(Dist. Surat), Gujarat, India. 2Assistant Professor, C.G Bhakta Institute of Biotechnology, Uka Tarsadia University, Mahuva Road,

Bardoli-394601, Surat (Dist. Surat), Gujarat, India.

*Corresponding Author: Email: [email protected]; [email protected]; Mob: +919448371037


ABSTRACT

Ginkgo biloba, a native of China, was probably a member of the mixed-mesophytic forest

community that once covered the hill country. In developing countries a large proportion of the

population relied heavily on traditional practitioners and medicinal plants to meet health care needs.

Growing interest in the field of Complementary and Alternative medicine places new obligations on

the clinicians. To assist in this process, a comprehensive review of Ginkgo biloba, the well studied

botanical medicine, was provided. Studies so far indicated the efficacy of Ginkgo biloba in tinnitus,

schizophrenia, psychotic organic brain syndrome, vertigo of undetermined origin. Also double blind

research showed Ginkgo biloba’s efficacy in conditions including cerebro-vascular insufficiency,

memory impairment, Alzheimer’s disease, multi-infarct dementia, resistant depression, peripheral

arterial insufficiency, venous insufficiency and asthma was summarized.

KEY WORDS: Ginkgo biloba, ginkgolides, dementia, Ginkgo extract

Review article

Cite this article:

Patel. H., Ingalhalli. R. (2013), A BRIEF REVIEW ON GINKGO BILOBA L.

(MAIDENHAIR TREE) - A RARE MULTIPURPOSE MEDICINAL PLANT,






INTRODUCTION

In developing countries a large proportion

of the population relies heavily on traditional

practitioners and medicinal plants to meet

health care needs (Gartoulla, 1993). Ginkgo

biloba tree is commonly known as maidenhair

tree. Since it has no close living relatives, it is

also called a living fossil. Ginkgo biloba is

cultivated nowadays in many parts of the world

as an ornamental plant and commercially

grown for its leaves which contain several

medicinal properties (DeFeudis, 1998). The use

of Ginkgo leaf extract for pharmaceutical

purposes was originally developed in Germany

in 1965, and the first commercially available

Ginkgo leaf extract was registered for human

use in 1974 in France, under the code-name

"EGb 761" (DeFeudis, 1998). Scientists from

various countries have been conducting clinical

trials to establish the efficacy of leaf extract in

treating tinnitus, vertigo, Alzheimer’s disease,

dementia, anxiety, cerebral dysfunction (Ref;

Table 1–5). Clinical trials have also been

counducted to find out the efficacy of leaf

extract to treat various other diseases (Ref;

Table 6). The Ginkgo leaf extract EGb 761 has

shown to have low acute and chronic toxicities

(Woerdenbag and De Smet, 2000). Ginkgo

biloba has proved its efficacy in treating a

considerable number of diseases and still

provides ample opportunities to future

researchers to unravel the hidden medicinal

properties. Considering the critically

endangered status of Ginkgo biloba in the wild,

it has become inevitable to cultivate it in

different parts for its utilization.

Habit

Ginkgo biloba is a dioecious tree, wind-

pollinated, animal-dispersed gymnosperm with

evolutionary affinities to both cycads and the

conifers which has been used in traditional

Chinese medicine for about 5000 years

(Saquires, 1999; Norstog et al., 2004). Ginkgos

are large trees, normally reaching a height of

20–35 m (66–115 feet), the tree has an angular

crown and long, somewhat erratic branches,

and is usually deep rooted and resistant to wind

and snow damage. Young trees are often tall

and slender, and sparsely branched; the crown

becomes broader as the tree ages (Royer et al.,

2003). The leaves are unique among seed

plants, being fan-shaped with veins radiating

out into the leaf blade, sometimes bifurcating

(splitting), but never anastamosing to form a

network. Two veins enter the leaf blade at the

base and fork repeatedly in two; this is known

as dichotomous venation. The leaves are

usually 5–10 cm (2–4 in), but sometimes up to

15 cm (6 in) long. The old popular name

"maidenhair tree" is because the leaves

resemble some of the pinnae of the maidenhair

fern, Adiantum capillus-veneris (Wang, 1961).

Distribution and status

As a wild species, Ginkgo biloba is native

to China and was probably a member of the

mixed-mesophytic forest community that once

covered the hill country bordering the Yangtze

River valley (He et al., 1997). The species is

highly adaptable and grows well in most

regions with a distinct seasonality and

moderate rainfall, including areas with warm-

temperate, cold-temperate, or Mediterranean

climates (Shen et al., 2004). For centuries, it

was thought to be extinct in the wild, but is

now known to grow in at least two small areas

in Zhejiang province in eastern China, in the

Tian Mu Shan Reserve. However, recent

studies indicate high genetic uniformity among

Ginkgo trees from these areas, arguing against

a natural origin of these populations and

suggesting the Ginkgo trees in these areas may

have been planted and preserved by Chinese

monks over a period of about 1,000 years

(Tang et al., 2012). The greater genetic

diversity has been found in Southwestern China

populations, in mountains surrounding eastern

Qinghai-Tibet Plateau (Tang et al., 2012; Fu et

al., 1999). Where it occurs in the wild, it is

found infrequently in deciduous forests and

valleys on acidic loess (i.e. fine, silty soil) with

good drainage. The soil it inhabits is typically

in the pH range of 5.0 to 5.5 (Barrett, 2004).

Medicinal importance

Extract of Ginkgo leaves contain

bioflavonoids (amentoflavone, bilobetin,

http://en.wikipedia.org/wiki/Leaf

http://en.wikipedia.org/wiki/Anastomosis

http://en.wikipedia.org/wiki/Zhejiang

http://en.wikipedia.org/wiki/Tianmu_Mountain

http://en.wikipedia.org/wiki/Loess



ginkgetin), flavonoids (quercetin,

isorhamnetins, kaempferol), proanthocyanidins,

Ginkgolides A,B,C, bilobalide (Barrett, 2004;

Kraft and Hobbs, 2004; Deng and Zito, 2003;

Evans, 2002; McKenna et al., 2001; Bruneton

2000; Samuelsson, 1999). Most Ginkgo

extracts available in the European market are

standardized to 24% flavones glycosides and

6% terpene lactones (Mahady et al., 2001;

Ahlemeyer and Krieglstein, 1998; Cott, 1995).

Although some Ginkgo preparations have also

been applied parenterally, the great are ingested

as tablets, capsules or extracts (Koltringer et

al., 1989). Leaf extract of Ginkgo is made from

dried Ginkgo leaves and has a standardized

content of 22–27% flavonol glycosides and 5–

7% terpene trilactones. This extract is taken

internally for the treatment of cerebral and

peripheral vascular diseases, as well as to

alleviate some of the ailments associated with

ageing, including dizziness, ringing in the ears,

and short-term memory deterioration

(DeFeudis, 1998; Juretzek, 1997). Ginkgo

supplements are usually taken in the range of

40–200 mg per day. Clinical trials to date have

utilized the standardized products EGb761 and

to a lesser extent LI 1370 (Bluementhal et al.,

2000). Details of clinical trials using Ginkgo

biloba are given in Tables 1 to 6.

Table-1: Selected Clinical Trials Employing Ginkgo on Dementia

Reference Plant part /

herbal product

Purpose of study Number

of

Subjects

Results

Ihl R et al., 2010 Ginkgo extract single dose

of 240 mg daily

Treatment of mild to

moderate dementia

Not

available

Effective

Mazza, M et al.,

2006

160 mg of ginkgo extract Treatment of dementia Not

available

Effective

Kanowski and

Hoerr, 2003

EGb 761 standardized

extract

Use of ginkgo

biloba extract for

the treatment of

dementia

Not

available

Effective

Van Dongen et

al., 2003

EGb 761Standardized

extract

Treatment for

dementia

214 Not

effective

Le Bars, et al.,

2002

EGb 761 Standardized

extract

Treatment for

dementia

Not

available

Effective

Le Bars et al.,

2000

EGb 761 Standardized

extract

Treatment for

dementia

309 Effective

Van Dongen et

al., 2000


extract

Treatment for

dementia

196 Not

effective

Kanowski et al.,

1996


extract

Treatment for

dementia

156 Effective

Le Bars et al.,

1997

Ginkgold (EGb 761

tablets)

Treatment for

dementia

202 Effective

Haase et al., 1996

EGb 761standardized

extract

Treatment for

dementia

40 Effective

Hofferberth,

1989

Tebonin forte (EGb 761)

Standardized extract

Treatment for

dementia

40 Effective



Table-2: Selected Clinical Trials Employing Ginkgo on Alzheimer's disease


herbal product

Purpose of study Number of

subjects

Results

DeKosky et al,

2008

Twice-daily dose

120 mg extract

Alzheimer's disease in

adults 75 years or older

Not available Not

effective

Stough et al.,

2001

EGb 761

Standardized extract

Memory

enhancement

61 Effective

Rigney et al.,

1999 Kaveri Memory

enhancement 31 Not

effective

Kanowski et

al., 1996

Ginkgo extract 120 mg

daily

Treatment of Alzheimer 216 Effective

Table-3: Selected Clinical Trials Employing Ginkgo on Asthma


herbal product


subjects

Results

Li et al., 1997

Concentrated leaf

liquid (Chinese product)

Treatment for asthma

61 Effective

Anonymous,

1989

Ginkgo extract Inhibitor of platelet

activating factor Not available Effective

Table-4: Selected Clinical Trials Employing Ginkgo on Cerebral Insufficiency


herbal product

Purpose of study Number

Of subjects

Results

Brautigam et al.,

1998

Geriaforce

(liquid extract)

Treatment for cerebral

insufficiency

197 Effective

(improvement of

short term visual

memory)

Vesper and

Hansgen, 1994

Kaveri LI 1370 Treatment for cerebral

insufficiency

86 Effective

Grassel, 1992

Rokan ® (EGb

761)


insufficiency

53 Effective

Bruchert et al.,

1991

Kaveri LI 1370 Treatment for cerebral

insufficiency

209 Effective

Schmidt et al.,

1991

Kaveri® LI 1370 Treatment for Cerebral

insufficiency

99 Effective

Eckmann, 1990

LI 1370

(standardized

liquid extract)


insufficiency

58 Effective



Table-5: Selected Clinical Trials Employing Ginkgo on Tinnitus


herbal product


subjects

Results

Raja et al.,

2004

Ginkgo biloba

Tablets

To evaluate the

efficacy of ginkgo

for the treatment

of tinnitus

66 Not effective

Drew and

Davies, 2001

LI 1370 standardized

extract

Treatment for

tinnitus

956 Not effective

Coles R. 1988 Ginkgo biloba extract

(40 mg three times a

day for 12 weeks)

Treatment of

tinnitus

21 Slight improvement

but treatment was

ineffective

Table-6: Selected Clinical Trials Employing Ginkgo on other diseases


herbal product


subjects

Results

Muir et al.,

2002

Seredrin

(standardized

Ginkgo biloba extract)

Treatment for

Raynaud’s

syndrome

Not

available

Effective

Prasad et al.,

2003

Ginkocer (Ginkgo biloba

extract)

Treatment for vitiligo 47 Effective

Gertsch et al.,

2002

Ginkgo biloba extract

Treatment for acute

mountain sickness

26 Effective (as

pretreatment)

Kang et al.,

2002

Ginkgo biloba extract

Treatment for

Antidepressant

induced sexual

dysfunction

19 Not effective

Chen et al.,

2003

Ginkgo biloba exocarp

polysaccharides

(GBEP) capsule

preparation taken

orally

To observe the

clinical efficacy of

(GBEP capsule

preparation in

treating upper

digestive tract

malignant tumors of

middle and late stage

86 The GBEP

Capsule preparation

had positive

therapeutic effects on

upper digestive

tract malignant

tumors of middle

and late stages

Brochet et al.,

1995

Intravenoous

application of

ginkgolide B

Treatment for

exacerbations of

multiple sclerosis

104 Not effective

Sikora ,1989 Ginkgo biloba extract 6o

mg per day for 12–18

months

Treatment of

impotence

60 Improvement in penile

arterial blood flow

Haguenauer,

1986

Ginkgo biloba extract Treatment of

idiopathic vertigo

70 Asymptomatic in 47%

patients

Lagrue et

al.,1986

Ginkgo biloba extract Treatment of cyclic

edema

10 Out of ten 3 women

showed complete

elimination of edema

and six were improved



The Special Ginkgo biloba leaf extracts are

generally well tolerated with low incidence of

side effects. Less than two percent of the

patients develop side effects and no serious side

effects have been noted in any trials. The side

effects include gastrointestinal complaints,

headache, sleep disturbances, dizziness and

allergic skin reactions. The ginkgo leaf extract

EGb 761 has been shown to have low acute and

chronic toxicities (Woerdenbag and De Smet,

2000).

Cultivation

Ginkgo has long been cultivated in China;

some planted trees at temples are believed to be

over 1,500 years old. In China, Ginkgo is

cultivated across a broad range of moisture,

temperature, and topographic gradients

between 25° and 42°N latitude, where

minimum winter temperatures can reach –32°C

and maximum summer temperatures 42°C (He

et al., 1997). The first record of Europeans

encountering it is in 1690 in Japanese temple

gardens, where the tree was seen by the

German botanist Engelbert Kaempfer. Because

of its status in Buddhism and Confucianism,

the Ginkgo is also widely planted in Korea and

parts of Japan; in both areas, some

naturalization has occurred, with ginkgos

seeding into natural forests. In some areas,

most intentionally planted Ginkgos are male

cultivars grafted onto plants propagated from

seed, because the male trees will not produce

the malodorous seeds. The popular cultivar

'Autumn Gold' is a clone of a male plant.

Ginkgos adapt well to the urban environment,

tolerating pollution and confined soil spaces

(Gilman, 2008).

They rarely suffer disease problems, even

in urban conditions, and are attacked by few

insects (Boland et al., 2002; Minnesota, 2008).

For this reason, and for their general beauty,

ginkgos are excellent urban and shade trees,

and are widely planted along many streets.

Ginkgos are also popular subjects for growing

as penjing and bonsai; they can be kept

artificially small and tended over centuries.

Furthermore, the trees are easy to propagate

from seed. In cultivation in the United

Kingdom G. biloba has gained the Royal

Horticultural Society's Award of Garden Merit.

The Ginkgo leaf is the symbol of the Urasenke

school of Japanese tea ceremony. The tree is

the national tree of China, and is the official

tree of the Japanese capital of Tokyo, and the

symbol of Tokyo is a ginkgo leaf (Minnesota,

2008).

CONCLUSION

With the growing patient demand for

alternative, complementary, natural, or

integrated approaches in treating disease, it is

increasingly important for clinicians to develop

referral relationships with well-trained

Complementary and Alternative Medicine

providers. Ginkgo biloba extract (GBE), one of

the most commonly used and best-researched

phytomedicines, has documented efficacy for

many conditions. Use of GBE in cerebro-

vascular insufficiency, memory impairment in

the elderly, Alzheimer’s disease, multi-infarct

dementia, resistant depression, peripheral artery

insufficiency, venous insufficiency and asthma

is well supported by multiple studies. GBE for

tinnitus, schizophrenia, psychotic organic brain

syndrome, vertigo of undetermined origin, and

PMS, although less supported, still deserves

serious consideration because of GBE’s high

tolerability, and the limited or complete lack of

efficacy with conventional treatments for these

conditions.

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http://www.sustland.umn.edu/design/insect.htm






EFFECT OF BADARA STEM BARK (ZIZIPHUS JUJUBA LAMB.) IN THE

MANAGEMENT OF VATAJA KASA IN CHILDREN

Chethan Kumar V K1, Shailaja U

2

1PhD Scholar & Associate Professor, Dept of Kaumarabhritya, S.D.M.College of Ayurveda, Udupi,

Karanataka. India 2Professor & Head, Dept. of Kaumarabhritya, S.D.M.College of Ayurveda & Hospital, Hassan, Karnataka,

India

*Corresponding Author: Email: [email protected]; Mob: +919986550745


ABSTRACT

Kasa (Cough) is one of the most frequently encountered problems in the Balyavastha (Pediatric

age). Recurrent attacks makes the school going child suffer and may have its adverse effects on the

studies of the child. The present clinical study was carried out to evaluate the effect of Syrup Badara

in Vataja kasa (Dry cough) in children. The present study was conducted on children presenting with

Vataja kasa in between the age group of 2–10 years. The clinical trial was conducted on 73 patients.

They were divided into two groups; each group consisted of minimum 30 patients. The divided

Groups A and B were given syrup Badara and placebo concentrated sugar solution respectively. The

results were analyzed statistically. The analysis suggested that Syrup Badara is effective in reducing

the signs and symptoms of Vataja kasa.

KEY WORDS: Kasa, Cough, Shushka kasa, Badara, Ziziphus jujuba

Research article

Cite this article:

Chethan K V K, Shailaja U(2013), EFFECT OF BADARA STEM BARK

(ZIZIPHUS JUJUBA LAMB.) IN THE MANAGEMENT OF VATAJA KASA IN CHILDREN,





INTRODUCTION

Respiratory system is one system which is

in continuous contact with the external

environment since birth until one’s lifetime, so

it is most vulnerable to infections and

considered as the prime victim of hyper

sensitization in most of the circumstances

(Atkins D, 2008). Thus Respiratory Tract

Infections (RTI) accounts to about more than

50% of patients attending pediatric OPD in

developing and even developed countries

worldwide. (Eggenberger K, 1993).

Pediatric age group is more vulnerable to

respiratory tract infections because of

anatomical and physiological peculiarities (i.e.

small narrow airways (Drake Lee A B, 1987),

hypertrophied lymphoid tissues (Hibber J,

1987), underdeveloped paranasal sinuses

(Diane E P, 2008), mucous hyper secretion and

peculiarities of Eustachian tube (Maw A R,

1987) and social factors such as attending

school, improper food and eating habits. As

cough is the most frequent symptom of

respiratory diseases (Innes J A, 2006) majority

of the patients present recurrent cough as the

manifestation of recurrent respiratory disease.

In Ayurvedic point of view descriptions of

disease Kasa clearly correlate with cough.

Moreover the patho-physiology (Samprapti) of

Kasa almost exactly correlates the mechanism

of cough reflex (Trikamji Yadavji, 2009).

Early intervention is necessary in case of

Kasa(cough) as it is a potential Nidanarthakara

Vyadhi (Diseases as causative factors for other

diseases)to produce Kshaya(consumption)

(Trikamji Yadavji, 2009). Also it is important

to treat any Balaroga (Pediatric disorder)at the

earliest as it may hamper the proper Vriddhi

(Growth and development) of a child which is

clearly described by Acharya Charaka, that

Avighata(Absence of inhibiting factors) as

shareera vriddhikara bhava (factors

responsible for growth of body) (Trikamji

Yadavji, 2009). Thus in the present work Kasa

(Cough) was taken as the subject of

intervention.

The Shamana (Palliative treatment) line of

treatment that includes oral administration of

medicine is of utmost importance as the

administration of syrup is very easy and also

effective compared to Shodhana (Purificatory

treatment) in children. Many research works

have been carried out in relation to the

Shamana treatment as directed in Ayurveda and

their therapeutic effect is proved. After critical

analysis of the drug ie. Stem bark of Badara

had been selected as the trial drug to evaluate &

establish its efficacy in combating the signs &

symptoms of Vataja Kasa (Dry cough).

METHODOLOGY

Present study was carried out for the

scientific understanding of this trial drug stem

bark of Badara (Ziziphus jujuba Lam) in the

management of Vataja Kasa. The present study

was conducted after the approval of the

Institutional Ethics committee with reference

IEC no – SDM/IEC/09/2008-2009. The study

was conducted on children of Vataja Kasa in

between the age group of 2–10 years. Patients

were divided into 2 groups. Group A and B

were treated with Syp. Badara and Syp. Sugar

Solution respectively. For preparing the syrup,

the following methods were followed at SDP

Remedies and Research Centre, Puttur D.K.,

Karnataka.

After thorough cleaning of stem bark of

Badara, it was powdered into a coarse form.

One part of the powder of the drug was

soaked with 8 parts of water and kept for

one night.

Next day the contents were boiled and

reduced to ¼ th

part and filtered.

In the obtained mixture 2 parts of sugar was

dissolved and re boiled to obtain one thread

consistency of the syrup and cooled to room

temperature.

Lastly it was filtered and sealed in sterilized

100ml bottles.

For the purpose of controlled study a

Placebo in the form of concentrated sugar

syrup of the same consistency was

prepared.



Stem bark of Badara was collected from

Kasargod District and authenticated by

Smt.Vidya, Botanist, SDP Remedies and

Research Centre, Puttur D.K., Karnataka.

The observations of the cases were

recorded according to the research proforma.

Criteria for selection of patients

Patients with signs and symptoms of

Vataja Kasa – viz., Shushka Kasa (Non

productive cough), Shirashoola (Head ache),

Swarabheda (Hoarseness of voice), Parshwa

Shoola (Pain in the flanks), Ura Shoola (Chest

pain), Shushka vaktra (Dryness of mouth),

Kantodhvamsanam (Throat irritation) were

selected randomly irrespective of their sex,

religion etc. from Kaumarabhritya O.P.D of

S.D.M. Ayurvedic Hospital, Hassan, Karnataka,

India

Inclusion criteria

Patients presenting with Vataja Kasa (Dry

Cough).

Any of two or more symptoms described in

the context of Vataja Kasa (Dry Cough).

Patients of Vataja Kasa (Dry Cough) with

chronicity of less than 15 days duration.

Patients irrespective of sex, religion,

socioeconomic status and between the age

group of 2–10yrs are taken.

Exclusion criteria

Other than Vataja Kasa (Dry cough).

Kasa as anubhandha lakshana

(Complication) in other systemic diseases.

Study design

For diagnostic purpose the signs and symptoms

mentioned below were taken for the study.

Shushka Kasa (Non productive cough)

Shirashoola (Head ache)

Swarabheda (Hoarseness of voice)

Parshwa Shoola (Pain in the flanks)

Ura Shoola (Chest pain)

Shushka vaktra (Dryness of mouth)

Kantodhvamsanam (Throat irritation)

Dose, duration and mode of administration:

Duration of the treatment was 10 days.

Group - A - Syp. Badara 5 ml for 5–10 years of

age group & 2.5 ml for 2–5 years of age group

in QID (6th

hourly) after food.

Group – B - Syp.Sugar Solution 5 ml for 5–10

years age group & 2.5 ml for 2–5 years of age

group in QID (6th

hourly) after food.

Criteria for assessment

Children suffering from Vataja Kasa (Dry

Cough) were assessed and evaluated on the

basis of the following parameters.

KasaVega (Bouts of cough)

Duration of each bout

Anidra (Sleeplessness)

Throat congestion

Absolute Eosinophil count AEC

TABLE 1: Registration data

Groups No of patients

Total Registered Drop out Completed

Group A

(Syp.Badara)

31 1 30

Group B

(Syp.Sugar solution)

42 12 30

Total 73 13 60



RESULTS & DISCUSSION:

Table no 2 indicates that Syrup Badara

provided relief in all the clinical symptoms of

Vataja Kasa (Dry cough). It provided 56.11%

relief in Shushka Kasa (Non productive cough),

82.69% relief in Shirashoola (Head ache),

66.67% in Swarabheda (Hoarseness of

voice),86.36% in Parshwa Shoola (Pain in the

flanks), 91.07% in Ura Shoola(Chest pain),

63.33% in Shushka vaktra (Dryness of mouth),

66.67% in Kantodhvamsanam (Throat

irritation), 57.22% in Kasa Vega (Bouts of

cough), 56.67% in duration of each bout of

cough, 80% relief in Anidra (Sleeplessness).

All the changes were statistically significant.

Table no 3 indicates that Syrup Sugar

solution provided relief in some of the clinical

symptoms of Vataja Kasa. It provided 23.33%

relief in Shushka Kasa (Non productive cough),

89.47% relief in Shirashoola (Head ache),

13.33% in Swarabheda (Hoarseness of voice),

17.39% in Parshwa Shoola (Pain in the flanks),

16.67% in Shushka vaktra (Dryness of mouth)

19.44% in Kantodhvamsanam (Throat

irritation), 25% in Kasa Vega (Bouts of cough),

24.72% in duration of each bout of cough,

25.93% relief in Anidra (Sleeplessness). All the

changes were statistically significant except in

Parshwa Shoola.

TABLE 2: Effect on symptoms, signs and hematological parameters of Vataja Kasa in Group A

Features N BT AT % of

relief

SE T P

Shushka Kasa-(Non productive cough)

30 1.93 0.90 56.11 0.03 31.0 <0.001

Shirashoola- (Head ache) 26 1.0 0.2 82.69 0.09 9.05 <0.001

Swarabheda- (Hoarseness of voice) 30 1.60 0.6 66.67 0.07 14.75 <0.001

Parshwa Shoola- (Pain in the flanks) 22 0.90 0.17 86.36 0.10 7.71 <0.001

Ura Shoola- (Chest pain) 28 1.03 0.13 91.07 0.06 16.16 <0.001

Shushka vaktra(Dryness of mouth) 30 1.70 0.7 63.33 0.05 20.86 <0.001

Kantodhvamsanam(Throat irritation) 30 1.63 0.53 66.67 0.05 20.86 <0.001

KasaVega (Bouts of cough) 30 1.93 0.87 57.22 0.05 23.03 <0.001

Duration of each bout 30 1.93 0.90 56.67 0.03 31.00 <0.001

Anidra (Sleeplessness) 30 1.57 0.40 80 0.07 16.86 <0.001

Rhonchi 8 0.40 0.13 75.0 0.08 3.25 <0.01

Throat congestion 30 1.1 0.23 81.67 0.06 13.73 <0.001

Total WBC 30 8912.2 8861.7 0.51 ↓ 15.49 3.26 <0.01

Neutrophil (%)

30 67.3 66.13 1.58 ↓ 0.49 2.37 <0.05

Lymphocyte (%) 30 26.2 28.27 9.94 ↑ 0.55 3.77 <0.01

Eosinophil (%) 30 6.2 5.6 8.79 ↓ 0.10 5.83 <0.001

Monocyte (%) 9 0.3 0 100 ↓ 0.9 3.53 <0.01

AEC 30 519.4 479.2 6.94 ↓ 14.29 2.81 <0.01

ESR (mm/1st hr) 30 10.47 8.67 18.99 ↓ 0.14 12.95 <0.001

Hb (gm %) 30 13.37 13.42 0.42 ↑ 0.04 1.19 >0.05

Note: ↓decrease , ↑increase ; N = number of frequencies, BT = Before Treatment AT = After Treatment; % = Percentage

SE = Standard Error; T = Student ‘T’ Test; P = Probability



TABLE 3: Effect on symptoms, signs and hematological parameters of Vataja Kasa in Group B

Features N BT AT % of relief SE T P

Shushka Kasa-(Non productive cough)

30 2.00 1.53 23.33 0.09 5.04 <0.001

Shirashoola- (Head ache) 19 0.70 0.20 89.47 0.12 4.01 <0.01

Swarabheda- (Hoarseness of voice) 30 1.73 1.43 13.33 0.10 3.07 <0.01

Parshwa Shoola- (Pain in the flanks) 23 0.83 0.77 17.39 0.10 0.70 >0.05

Ura Shoola- (Chest pain) 23 0.80 0.97 2.17 ↑ 0.08 1.98 >0.05

Shushka vaktra (Dryness of mouth) 30 1.67 1.30 16.67 0.10 3.61 <0.01

Kantodhvamsanam (Throat irritation)

30 1.53 1.10 19.44 0.10 4.18 <0.01

KasaVega (Bouts of cough) 30 2.20 1.63 25 0.09 6.16 <0.001

Duration of each bout 30 2.17 1.57 24.72 0.09 6.60 <0.001

Anidra (Sleeplessness) 27 1.27 0.97 25.93 0.12 2.52 <0.05

Rhonchi 11 0.53 0.30 50 0.09 2.54 <0.05

Throat congestion 27 0.93 1.0 7.14↑ 0.05 1.44 >0.05

Total WBC 30 8938.9 8946.77 0.12↑ 9.38 0.84 >0.05

Neutrophil (%)

30 67.93 68.6 1.14↑ 0.48 1.40 >0.05

Lymphocyte (%) 30 25.53 25.10 0.46↓ 0.53 0.82 >0.05

Eosinophil (%) 30 6.37 6.40 0.83↑ 0.06 0.57 >0.05

Monocyte (%) 5 0.17 0 100 0.07 2.41 <0.05

AEC 30 549.10 550.87 0.75 ↑ 2.90 0.61 >0.05

ESR (mm/1st hr) 30 8.93 9.07 6.11↑ 0.29 0.45 >0.05

Hb (gm %) 30 12.83 12.62 1.63↓ 0.04 4.75 <0.001

Note: ↓decrease , ↑increase ; N = number of frequencies, BT = Before Treatment AT = After Treatment; % =

Percentage; SE = Standard Error; T = Student ‘T’ Test; P = Probability

Table no 4 gives a clear picture of

comparison between the two groups. Group A

showing highly significant result i.e p <0.001

in all symptoms of Vataja Kasa (Dry cough)

except Shirashoola (Headache).

Syrup Badara helps in reducing

Shirashoola (Headache), Parshwa Shoola (Pain

in the flanks) and Ura Shoola (Chest pain) due

to Ushna Veerya (Hot potency) of the drug

Badara which acts as Vedana sthapaka

(Analgesics).

Kasa vega (Bouts of cough), Shushka kasa

(Non productive cough), Shushka vaktra

(Dryness of mouth) and Swara bheda

(Hoarseness of voice) are due to Rooksha Guna

(Dry quality) of Vata and the Snigdha guna



(Unctous quality) of Badara might provide

better relief in those symptoms.

Kashaya Rasa (Astringent taste) of stem

bark of Badara in Syrup Badara might do the

mucosal modulation, which cuts off the contact

of antigen (allergen/ pathogen) and receptors

(such as Dendritic Cells, MHCs those are

embedded in the mucosa of nose, pharynx,

larynx etc. This drug action explains the

symptomatic relief from Kasa (Cough).

TABLE 4: Comparative effect on symptoms, signs and hematological parameters of Vataja

Kasa in Group A and Group B

Note: ↓decrease, ↑increase and N.S not significant; N = number of frequencies, % = Percentage; T = Student

‘T’ Test; P = Probability

Marked decrease in ESR and simultaneous

decrease in AEC clearly indicates the drug’s

action on allergic conditions. Decrease in Total

counts and polymorphs indicate the drug’s

effectiveness even in infectious conditions.

The present study results goes very well

with the study conducted by Rakesh Johri K,

(1992) which indicates that the Stem bark of

Badara (Ziziphus jujuba Lamb) contains

abundant tannins, which exhibits surface action

on the pharyngo-laryngeal mucosa and

modulate them to decrease exudation, provides

a shield against the contact of any antigen

(pathogen/allergen), in addition tannins also

have local antimicrobial action which destroys

the microbes coming in contact.

Measures Group A Group B T p value

N % Relief N % Relief

Shushka Kasa-(Non productive cough) 30 56.11 30 23.33 5.80 < 0.001

Shirashoola- (Head ache) 26 82.69 19 89.47 0.71 N.S

Swarabheda- (Hoarseness of voice) 30 66.67 30 13.33 6.63 < 0.001

Parshwa Shoola- (Pain in the flanks) 22 86.36 23 17.39 7.23 < 0.001

Ura Shoola- (Chest pain) 28 91.07 23 2.17 ↑ 16.64 < 0.001

Shushka vaktra(Dryness of mouth) 30 63.33 30 16.67 6.08 < 0.001

Kantodhvamsanam(Throat irritation) 30 66.67 30 19.44 6.0 < 0.001

KasaVega (Bouts of cough) 30 57.22 30 25.0 6.12 < 0.001

Duration of each bout 30 56.67 30 24.72 6.23 < 0.001

Anidra(Sleeplessness) 30 80 27 25.93 7.14 <0.001

Rhonchi 8 75 11 50 1.57 N.S

Throat congestion 30 81.67 27 7.14↑ 11.80 <0.001

Total WBC 30 0.51 30 0.12↑ 2.95 < 0.01

Neutrophil (%) 30 1.58 30 1.14↑ 2.64 < 0.01

Lymphocyte (%) 30 9.94↑ 30 0.46 3.16 < 0.01

Eosinophil (%) 30 8.79 30 0.83↑ 5.27 < 0.001

Monocyte (%) 9 100 5 100 0 N.S

AEC 30 6.94 30 0.75↑ 3.08 < 0.01

ESR (mm/1st hr) 30 18.99 30 6.11↑ 4.73 < 0.001

Hb (gm %) 30 0.42↑ 30 1.63↓ 4.28 < 0.001



CONCLUSION

Kasa (Cough) being a common childhood

ailment has been given more emphasis by the

medical people because it diverts parents

psyche from their routine work. All the changes

were statistically highly significant i.e p

< 0.001. Syrup Badara also showed statistically

highly significant relief in duration of each

bout, frequency of bouts, throat congestion,

AEC and also show improvement in Hb gram

%. The findings suggest that the Syrup Badara

is an effective and safe herbal formulation for

the Vataja Kasa (Dry cough).

REFERENCES

Atkins,D. (2008). Diagnosis of allergic

diseases. In: Behrman , RE.(ed.)

Nelson’s Textbook of Pediatrics.

Philadelphia, Saunders Elsevier. Ch

140, p.938

Diane, EP. (2008). Sinusitis. In : Behrman, RE.

(ed.) Nelson’s Textbook of Pediatrics.

Philadelphia, Saunders Elsevier. Ch

377, p.1749.

Drake Lee, AB. (1987). The catarrhal child. In

: Kerr, AG. (ed.) Scott. Brown’s

otolaryngology. Butterworths & Co.

Ltd, London. p. 280–290

Eggenberger, K. (1993). Respiratory tract

infections most frequently seen in

pediatric outpatient care. Ars Medicine.

1 May, p. 24–40.

Hibbert, J. (1987) Tonsils and Adenoids. In :

Kerr, AG. (ed.) Scott. Brown’s

Otolaryngology. Butterworths & Co.

Ltd, London. p. 368–383.

Innes, JA. (2006). Respiratory disease. In :

Davidson, S S. (ed.) Davidson’s

Principle and Practice of Medicines.

Churchill Livingstone Elsevier,

Philadelphia. p.657

Maw, AR. (1987). Otitis media with effusion.

In : Kerr, AG. (ed.) Scott.I,

Butterworths & Co. Ltd, London. p.

157–176.

Rakesh Johri, K. (1992). Piperine-mediated

changes in the permeability of rat

intestinal epithelial cells. The status of

γ-glutamyl transpeptidase activity,

uptake of amino acids and lipid

peroxidation. Biochemical

Pharmacology. 1 April, p1401–1407.

Trikamji Yadavji, (2009) Kasa Chikitsa

Adhyaya., Charaka Samhita,

Chaukhambha Sanskrit Sansthan,

Varanasi. p. 540, p. 227, p. 332





EFFECT OF PARNAYAVANI (COLEUS AMBOINICUS LOUR.) ON MES

INDUCED EPILEPSY IN RATS

Sharma Monica1*, Khemani Naresh

2, Singh Janardhan

3

1Lecturer, Department of Dravya Guna, CBPACS, Najafgarh, New Delhi, India

2Professor & HOD, Department of Dravya Guna, NIA, Jaipur, India

3Department of Pharmacology, PGIMS, Rohtak, Haryana, India

*Corresponding Author: Email: [email protected] ; Mob: +91 7838241559


ABSTRACT

Epilepsy is considered as the most dreadful disease & it’s prevalence has been estimated at 5–10

person per 1000.The present study was designed to investigate the antiepileptic potential of Coleus

amboinicus Lour. on Maximal electroshock (MES) induced seizure model in rats. Total 50 Albino

rats of wistar strain were used in the study. Animals were divided into 5 groups each having 10 rats.

Fresh juice of Parnayavani i.e. Coleus amboinicus (2 ml/rat) was given in one group while the

other group receives Aqueous & alcoholic extract of Parnayavani (500 mg/bw p.o.). Phenytoin

(25 mg/kg bw i.p.) served as standard drug for comparision where as control group receives distil

water as vehicle. In MES model delay in HLE was taken as end point. C. amboinicus leaf juice

(CALJ) & alcoholic extract (CAalE) was having highest efficacy as antiepileptic drug in comparison

to its aq. extract. in MES model . Thus we observed that leaf juice as well as alcoholic extracts

of Coleus amboinicus possess anticonvulsant and neuroprotective activity.

KEYWORDS: Epilepsy, MES, Parnayavani, seizures.

Research article

Cite this article:

Sharma Monica, Khemani Naresh, Singh Janardhan (2013), EFFECT OF PARNAYAVANI

(COLEUS AMBOINICUS LOUR.) ON MES INDUCED EPILEPSY IN RATS,





INTRODUCTION

Humanity is suffering from various

psychosomatic disorders like anxiety,

depression, epilepsy, dementia, Parkinson’s

(Nayak AP, 2010) and Alzheimer’s inspite of

great advances in the field of medical sciences.

Epilepsy is considered as the most dreadful

disease amongst all, which continues to effect

human beings since ancient time It’s

prevalence has been estimated at 5–10 person

per 1000 and it is considered as the second

most common neurological disorder next to

stroke (Katzung BG, 2007). The contemporary

system of medicine has ample of potent

antiepileptic remedies but they also have a risk

of developing vast array of side effects

including chronic toxicity, teratogenicity,

adverse effect on cognition and behaviour etc.

(Raza MF et al.,2001). Amid of all these, it is

the high time to search remedies from the

traditional treasure which may be proven as

safe & effective antiepileptic agent.

Coleus amboinicus Lour. Synonym C.

aromaticus Benth. or Plectranthus amboinicus

Lour. belonging to family Lamiaceae is

commonly known as Parnayavani in Sanskrit,

Patta ajwain in hindi, Karpurvalli in south

India & Country borage in English. It is a large,

succulent, aromatic herb found throughout in

India and is native of South and East Africa.

The leaves of this plant are traditionally used

for the treatment of severe bronchitis, asthma,

diarrhoea, epilepsy, renal & vesicle calculi &

fever (Bhakuni DS et al., 1969). It has been

reported to exhibit antilithotic,

chemopreventive & antioxidant properties

(Padma, P.R. et al., 1998). Although

Parnayavani have been in folklore use in

Epilepsy no scientific investigation is yet to be

done for establishing its antiepileptic activity.

Therefore the present study aimed to study the

effect of aqueous & alcoholic extract and leaf

juice of Coleus amboinicus Lour. (CAaqE),

(CAalE), (CALJ) which are known to have

anti-epileptic property on animal model.


The study was conducted in the P.G. deptt.

of Dravyaguna, NIA, Jaipur & deptt. of

Pharmacology, Pt. B.D.Sharma Post Graduate

Institute of Medical Sciences, Rohtak. Total 50

Rats were taken for the study.

Collection of Plant Material:

The leaves of Coleus amboinicus were

collected from the medicinal herbal garden of

NIA Jaipur where it grows. The Botanical

identification was carried out by BSI, Jodhpur

letter no.BSI/AZC/A.19014/SE-1/Estt./162

dt.23.6.2010. Leaves were dried under shade,

coarsely powdered & were packed separately in

air tight containers.

Extract Preparation :

For each extraction about 200 g powdered

drug was packed in Soxhlet apparatus. About

1.5 litres of solvent i.e. ethanol for alcoholic

extraction & distilled water for aqueous

extraction, was placed in a round bottom flask

and a reflux condenser was attached above the

soxhlet. The solvent was heated to boil on

heating mantle & was subjected to extraction

for 12 h. The filtrate was evaporated to dryness

by keeping it on a water bath at 50–60˚C. This

process is repeated thrice to get the required

amount of extracts of Coleus amboinicus

(ethanolic & aqueous extract).

Preparation of leaf juice:

About 500 g of fresh leaves of Coleus

amboinicus were cut into small pieces & juice

was prepared by crushing them in a mortar &

pestle & by adding 30 ml distil water to it.

EXPERIMENTAL ANIMALS

Adult Albino rats of Wistar strain

(weighing b/w 100–180 g) of either sex were

used in the study. Rats were procured from the

disease free animal house, Haryana

Agricultural University (HAU), Hissar. The

experimental rats were housed in poly

propylene cages under laboratory conditions of

28 ± 2˚C temp with 75% relative humidity and



photoperiod of 12 h light & dark cycle. The rats

were given standard pellet diet supplied by

Hafed, India Limited, Rohtak) and water ad

libitum, throughout the experiment.

The protocol of the study was approved by

Institutional Ethical Committee (IAEC,

PGIMS, Rohtak) & the experiments were

carried out as per ethical guidelines for animal

protection and welfare bearing the CPCSEA

438/01/a/cpcsea/dt 17.07.2006 in its resolution

No: 9/IAEC/SVU/2006/dt 04.03.2006.

Chemicals:

Phenytoin (Epsolin, Cadilla, India) was

used in the study. All other chemicals used

were of analytical grade.

Selection of Doses:

As per OECD (2002) guideline no. 425, the

LD 50 of Coleus amboinicus was estimated to

be >5000 mg/kg. Hence, 1/10 th

of the LD 50,

i.e. 500 mg/kg, dose was selected for the study.

Fresh leaf juice was administered at a rate of

2 ml/rat (Baskar R. et.al., 1992).

Administration of test substance:

The test drugs were prepared by dissolving

the required extracts in distilled water. The

volume of administration was kept at 1 ml/kg

bw. A gastric catheter was used for oral drug

administration. Phenytoin was dissolved in

normal saline before i.p. administration.

Induction of Epilepsy

Seizures were induced by MES methods.

Experimental design (Study Protocol)

In the present study Method of Tomar &

Everett (1974) & (Cashin et al., 1962) was

used. Electric shock seizures were produced by

delivering a current of 150 mA through corneal

electrode for a period of 0.2 sec. from an

electro-convulsiometer. Animals which showed

tonic extension of hind limb were selected &

given overnight rest. On the next day the

animals were divided into 5 groups of 10

animals each.

Group 1- Served as Control & receives normal

saline (10ml/kg) as Vehicle.

Group 2- Receives Fresh leaf juice of Coleus

amboinicus (CALJ) (2 ml/rat).

Group 3- Receives aqueous extract of Coleus

amboinicus (CAaqE) (500 mg/kg bw p.o.)

Group 4- Receives alcoholic extract of Coleus

amboinicus (CAalE) (500 mg/kg bw p.o.)

Group 5- Receives Phenytoin (25 mg/kg bw

i.p.) standard drug for comparison.

Animals of Group 2 were pretreated with

leaf juice of Parnayavani (Coleus amboinicus)

electroshocks were given after 60 min of drug

administration. Animals of Group 3 & 4

received aqueous and alcoholic extract of

Coleus amboinicus respectively & again

electroshocks were given after 60 min of drug

administration. Group 1 and 5 were control &

standard drug group respectively. Animals

were considered protected if the drug prevents

or delays the appearance of hind limb extension

component of the Seizures. Effect of extracts

on different stages of convulsions i.e. hind limb

extension, flexion & clonic convulsions were

also noted. Recovery & death of Rats were also

recorded in each group.

Statistical Analysis

All the results were expressed as mean ±

standard error of mean (SEM) and analyses of

variance (ANOVA) for the data were

calculated by using the SPSS (Statistical

Package for Social Sciences) software.

RESULTS

Phytochemical screening

The preliminary phytochemical analysis

showed presence of alkaloids, carbohydrates,

glycosides, proteins, amino acids, flavonoids,

quinine, tannins, phenolic compounds and

terpenoids.



Effect of CALJ, CAaqE and CAalE on MES

induced epilepsy

The duration of tonic HLE in animals

treated with vehicle i.e. in control group was

10.1 ± 0.53 seconds. The CAaqE treated group

showed significant reduction in duration of

HLE (2.3 ± 0.99) whereas CALJ, CAalE and

standard drug Phenytoin treated group exhibit

completely abolished HLE phase as compared

to control group.The CALJ, CAaqE,CAalE and

phenytoin treated group have shown 100%,

60%,100% and 100% protection respectively

(Table-1).

Table No. 1. Effect of Coleus amboinicus leaf juice (CALJ), aqueous extract (CAaqE) &

alcoholic extracts (CAalE) on MES induced Seizures in rats.

Grou

p

Design of

treatment

n= 10

Dose mg/kg

Duration (seconds) in various phases

of convulsions

Mortality

%

Protecti

on Tonic

flexion

HL

extension

Clonus

I. Control - 3.6 ±

0.34

10.1±0.53 5.6±0.58 2/10 -

II. CALJ 2ml/rat p.o. 4.0±0.49 NIL*** 10.5±0.84** 0/10 100%

III. CAaqE 500 mg/kg

p.o.

4.1±0.433 2.3±0.99*** 12.7±0.817*

**

0/10 60%

IV. CAalE 500mg/kg

p.o.

3.2±0.388 NIL*** 14.7±1.69**

*

0/10 100%

V. Phenytoin 25mg/kg

i.p.

1.9±0.233

*

NIL*** 2.3±0.335 0/10 100%

Data represented as mean+ SEM of 10 rats. . Data compared with one way ANOVA

P <0.05 non significant **

P <0.01 significant ***

P <0.0001 highly significant

DISCUSSION

The MES induced seizures test is the most

validated experimental method for assessment

of antiepileptic drugs effective in generalised

tonic clonic seizures (Loscher W, Schmidt D

1988) (Oliveira FA et.al., 2001). The MES

model is used to identify compounds which

prevents seizures spread (Kupferberg HJ 1989)

(Stables JP, Kupferberg HJ 1995) and most of

these compounds have the ability to inactivate

voltage dependant Na+ channels in a dose

dependant fashion. Such compounds suppress

sustained repetitive firing in cultured neurons

(Mcnamara J, 2001). In the present study

CALJ, CAaqE, CAalE significantly inhibited

the tonic HLE in MES test . It suggests the

presence of antiepileptic compounds and their

potential utility in the management of

generalized tonic clonic seizures. Further CALJ

and CAalE were equally potent antiepileptic

agents in comparison to CAaqE. Hence CALJ,

CAaqE, CAalE may be expected to have

similar type of mechanism.

Carvacrol & Thymol (main active

constituents of Coleus amboinicus (Haque IU,

1988), (Pino J et al., 1989) belongs to the class

of monoterpenes. Terpinoids, the oxygenated

derivatives of terpenes are reported to exhibit

antioxidant properties (J. Grabmann, 2005),

(Johanna Grassmann et al., 2002). There is

convincing evidence that in seizures, level of

secondary metebolites of Lipid peroxidation

(LPO) i.e. Thio barbituric acid reactive

substances (TBARS) was significantly

increased & antioxidant drugs facilitate the

suppression of LPO enhancement (Bashkatova



V et al., 2003). Coleus amboinicus is reported

to have antioxidant property (Rao BS et al.,

2006) (Kumaran A et al., 2006). Thus it might

be suggested that the suppression of LPO

enhancement may be involved in the

mechanism of action of antiepileptic potential.

CONCLUSION

Thus we conclude that leaf juice as well as

alcoholic extract of Coleus amboinicus possess

anticonvulsant and neuroprotective activity and

thus can be effectively used for the treatment of

epilepticseizures.

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Bashkatova V, Narkevich V, Galina Vitskova

G and Vanin A (2003) . The influence

of anticonvulsant and antioxidant drugs

on nitric oxide level and lipid

peroxidation in the rat brain during

penthylenetetrazole - induced

epileptiform model seizures.Progress in

Neuro - Psychopharmacology &

Biological Psychiatry. Vol. 27, (3): p-

487–92.

Baskar R, Varalaksmi P and Amsaveni (1992).

Changes in tissue enzymes produced by

Coleus aromaticus experimental

Urolithiasis. Indian Drugs 29 (6): p-

254–58

Bhakuni DS, Dhar ML, Dhar MM, Dhawan

BN, Mehrotra BN (1969). Screening of

Indian plants for Biological activity part

II. Indian J Exp Biol 7: p-250–62.

Cashin CH,Jackson H. (1962). An apparatus for

testing anticonvulsant drugs by

electroshock seizure in mice. J

Pharmacol ;14: p-445–75.

Haque IU (1988). Analysis of volatile

constituents of Coleus aromaticus.

Journal of the Chem. Society of

Pakistan. 10 (3): p-369–71.

J. Grabmann (2005) .Terpenoids as Plant

Antioxidants. Vitamins & Hormone.

Vol 72: p-505–535.

Johanna Grassmann, Susanne Hippeli and

Erich F. Elstner (2002). Plant’s defence

and its benefits for animals and

medicine: role of phenolics and

terpenoids in avoiding oxygen stress.

Plant physiology & biochemistry Vol

40 issue 6-8, p- 471–478.

Katzung BG (2007). In “Basic & Clinical

Pharmacology”, 10th

Ed. Mc-Graw Hill,

Boston, Toronto; p-374–94.

Kumaran A and Karunakaran Joel R (2006).

Antioxidant and free radical scavenging

activity of an aqueous extract of Coleus

aromaticus. Food Chemistry. 97: p-

109–14.

Kupferberg HJ (1989). Antiepileptic drug

development program: a cooperative

effort of government and industry.

Epilepsia,30 (1); p-S51–S56.

Loscher W, Schmidt D (1988). Which animal

models should be used in the search for

new antiepileptic drugs? A

proposalbased on experimental and

clinical consideration. Epilepsy Res., 2;

p-145–181.

Mcnamara J (2001). Drugs effective in the

therapy of epilepsy, chapter 21 In:

Goodmann and Gilmann’s the

pharmacological basis of therapeutics.

Hardman GJ et. Al. Mcgrew hill

company Inc. Newyork , p-521–47.



Nayak AP (2010). Parkinsonism in Ayurvedic

perspective, A bird’s eye view. Global

J. Res.Med.Plants & Indigen. Med. 1

(11): p-629–38.

OECD (2002). Acute oral toxicity. Acute oral

toxic class method guideline 425

adopted 23.03.1996. In: Eleventh

Addendum to the, OECD, guidelines

for the testing of chemicals organisation

for economical co-operation and

development, Paris, June, 2000.

Oliveira FA, Almeida RN, Sousa MFV,

Barbosa-Filho JM, Diniz SA, Medeiros

IA (2001).Anticonvulsant properties of

Nsalicyloyltryptamine in mice.

Pharmacol Biochem Behav. 68; p-199–

202.

Padma, P.R., Bhuvaneshwari, V and

Silambuchelvi, K (1998). The activities

of enzymatic antioxidants in selected

green leaves. Indian J Nutr Dietet 35;

:p-1–3.

Pino J, Rosado A and Borges P (1989).Volatile

components in the essential oil of wild

oregano (Coleus amboinicus Lour.).

Food / Nahrung. 34 (9): p-819–23.

Rao BS Shanbhoge R, Upadhya D, Iagetia GC,

Adiga SK, Kumar P, Guruprasad K, and

Gayathri P (2006). Antioxidant,

anticlastogenic and radioprotective

effect of Coleus aromaticus on Chinese

hamster fibroblast cells (V79) exposed

to gamma radiation. Mutagenesis. 21

(4): p-237–42.

Raza MF, Shaheen MI, Choudhary A, Suria

AU, Rahman S, Sombati and Delorenzo

RJ (2001). Anticonvulsant activities of

the FS-1 Sub-fraction isolated from

roots of Delphinium denudatum.

Phytother. Res., 15; p- 426–430.

Stables JP, Kupferberg HJ (1995). The NIH

Anticonvulsant Drug Development

(ADD) Program: Preclinical

Anticonvulsant Screening project. In:

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RH, Mattson RH, Meldrum BS, Raven

Press, New York.p- 4–17.

Toman JEP,Everett GM (1974). In Evaluation

of drug activit Pharmacometrics.

Laurence DR & Bachard AL,Eds.

Academic press, London; p-287.





ANTIMICROBIAL POTENTIAL OF THE WILD AND CULTIVATED

VARIETY OF ERANDA (RICINUS COMMUNIS LINN.) ROOT

Doshi Krunal A1*, Sagar Dhwani H

2, Acharya R N

3

1Ph D Scholar, Department of Dravyaguna, IPGT&RA, GAU, Jamnagar.Gujarat, India

2 Head, Department of Microbiology, IAPS, GAU, Jamnagar, Gujarat, India

3 Associate professor, Department of Dravyaguna, IPGT & RA, GAU, Jamnagar, Gujarat, India.

*Corresponding author: [email protected]; Mob : 09898907572


ABSTRACT

Methanolic extracts of wild and cultivated variety of Eranda (Ricinus communis) roots were

evaluated for their antimicrobial activity against pathogenic microorganisms such as Escherichia

coli, Salmonella paratyphi, Staphylococcus aureus, Staphylococcus epidermidis, and Aspergillus

niger using agar well diffusion method. Cultivated variety showed additional antimicrobial activities

than wild variety. Wild root extract at 0.282 mg/ml showed inhibiting growth of E.coli and S.

epidermidis and cultivated root extract at 0.224 mg/ml showed inhibiting growth of E. coli and

Salmonella paratyphi. These findings established the potential of the cultivated variety of Eranda

root as an effective antimicrobial agent against selected organisms. However, further studies are

needed to evaluate active compounds and probable medicinal benefits in humans by clinical trials.

KEYWORDS: Castor oil plant, Eranda root, Ricinus communis, Methanolic extracts,

Research article

Cite this article:

Doshi K. A., Sagar D. H., Acharya R. N., (2013), ANTIMICROBIAL POTENTIAL OF THE WILD

AND CULTIVATED VARIETY OF ERANDA (RICINUS COMMUNIS LINN.) ROOT,





INTRODUCTION

For the treatment of infections occurred by

bacteria a wide variety of antibiotics are

commonly used (Tumah H, 2005). Due to

excess use of existing antimicrobial drugs,

multiple drug resistance has been developed.

Antimicrobial resistance is an alarming

condition for mankind, because most of the

infectious bacteria have developed multiple

drug resistance (Saeed S et al., 2007). In

concern to drawbacks of conventional

medicine, the use of natural products as an

alternate to the conservative treatment in

healing and cure of various diseases has been

raised in the last few decades (Saeed S and

Tariq P, 2007).

Ricinus communis Linn (Euphorbiaceae),

commonly known as Eranda in Ayurveda

(Kirtikar KR & Basu, 1985), is a soft-wooded

small tree widespread throughout tropics and

warm temperate regions of the world. It is

cultivated widely in India, especially in the

region of Saurashtra of Gujarat for seed oil. In

Ayurveda, the leaf, root and seed oil of this

plant have been used for the treatment of

inflammation and liver disorders (Kirtikar KR

& Basu, 1985), roots are indicated in

rheumatism, inflammation, backache, diseases

of abdomen, fever etc (API, 1999). It is

reported that this plant possesses

hepatoprotective (R Yanfg LL et al., 1987 and

Visen P et al., 1992), antidiabetic (Shokeen P et

al.,2008), laxative (Capasso F, 1994), and

antifertility (Sandhy K & Bobby RG, 2003)

activities. Methanol extract of root showed

Anti- inflammatory and Free radical

Scavenging activity (Ilavarasan R et al., 2006).

Methanol extracts of root also reported for its

antimicrobial activity (Abhishek M. et al.,

2012).

Nowadays, due to lack of sufficient

quantity of wild variety, the cultivated variety

is mainly used to full-fill the market demand of

its root. T antimicrobial effect of Eranda root

extract have been published. But no research

has yet been carried out to evaluate the

comparative antimicrobial effect of wild and

cultivated root extracts. Hence, both the

varieties were compared for their antimicrobial

activity.


Collection of Drugs

Fresh roots of wild (more than six months)

and cultivated variety (six months old) were

collected after proper identification of the plant

as Ricinus communis Linn. (Euphorbiaceae),

from the adjacent area of Jamnagar town of

Gujarat, India, with the help of a taxonomist

and a specimen (no. 1491wild/cultivated) of the

two were preserved in the department, for

further reference. The obtained roots were

shade dried and made into coarse powder with

the help of a mechanical grinder (Mess no 20).

Preparation of methanol extracts

Both the samples were subjected for

extraction with methanol (Shukla V.J. and

Bhatt U.B., 2001). The concentration of both

extracts for antimicrobial activity was 15

mg/ml.

Test microorganisms

The antibacterial experimental organisms

were Escherichia coli, Salmonella paratyphi

(Gram negative), Staphylococcus aureus,

Staphylococcus epidermidis (Gram positive).

The antifungal experimental organism was

Aspergilus niger. Both bacterial and fungal

strains were provided by by IMTECH,

Chandigarh, India and collected from MBRC,

Jamnagar. Antimicrobial assay was performed

by inoculation of sub cultured pathogenic

strains in nutrient broth. The experimental work

was carried out at Dept. of Microbiology,

IAPS, GAU, Jamnagar.

Determination of Minimum Inhibitory

Concentration (MIC)

MIC of the extracts against the above

mentioned organisms were determined by using

broth dilution technique. Different sets

containing a range of serial dilution of extracts



were prepared for the selected organisms. 3 ml

of nutrient broth was filled in each test tube and

autoclaved. Test tubes were left to cool down at

room temperature. In each set 200 µl of

antimicrobial extract was serially diluted in 13

test tubes and one test tube was kept as blank.

Test tubes were inoculated with different

pathogenic strains except blank. The inoculated

sets were kept for incubation at 37ºC for 24

hours. Optical densities of each were measured

at 600 nm. (Phillips R and Martyn R., 1999)

Antimicrobial Assay

Antimicrobial assay was carried out by agar

(well) diffusion method (Jahir Alam Khan et

al., 2011). Sterile nutrient agar plates

(containing 20 ml N-agar) were prepared. 300

μl of the selected pathogenic strains were

spreaded on different N-agar plates with the

help of L-shape spreader. 5 wells of 9 mm

diameter were bore with the help of sterile

borer. 150 µl and 200 µl of each wild root and

cultivated root extract were taken for the assay.

The antibacterial drug Gentamycin (80 µg/ml)

and antifungal drug Fuconozocon DT (80

µg/ml) were taken as standards. Plates were

incubated at 37ºC for 24 hours and observed

for zone of inhibition on next day. All

susceptibility tests were performed in

triplicates.

RESULT AND DISCUSSION

Upon performing minimum inhibitory

concentration (MIC) of methanolic extract of

wild sample and cultivated sample against the

above mentioned two gram positive and gram

negative organisms, the minimum

concentration of the wild variety sample extract

required to inhibit growth of E.coli and S.

epidermidis was 0.282 mg/ml, while inhibition

of S. aureus and S. paratyphi were not

observed in the concentration range selected for

the assay. Minimum concentration of cultivated

root extract required to inhibit growth of E. coli

and Salmonella paratyphi was 0.224 mg/ml

while no remarkable inhibition of the other

bacteria was observed (Table 1, Graph 1, Table

2 and Graph 2).

Upon performing the Agar diffusion

method, 150 µl of wild root extract showed

inhibition against S. epidermidis (1.1 mm zone)

and A. niger (1 mm zone) where as zone

measuring 1 mm was obtained against E. coli

when 200 µl of the same extract was taken.

Similarly 150 µl cultivated variety root extract

showed inhibition against A. niger (1 mm zone)

and 200 µl of this extract showed zone of

inhibition against all the remaining organisms.

The zones of inhibition of the extracts were

compared to that of the standards where it was

found that the zone obtained with standards

were larger than both the extracts. (Table 3 and

figure 1).

Table 1: MIC of wild root extract on different bacteria

Sr

no.

concentration of wild root sample

extract

(µg/ml)

O.D. (Optical Density) at 660 nm on different

organism

E.coli Salmonella

paratyphi

S.

aureus

S.

epidermidis

1 0.00 0.20 0.16 0.38 0.30

2 14.10 0.18 0.15 0.37 0.29

3 56.40 0.17 0.14 0.35 0.25

4 84.60 0.16 0.14 0.34 0.17

5 126.90 0.16 0.13 0.30 0.14

6 169.20 0.15 0.12 0.29 0.07

7 282.00 0.12 0.10 0.27 0.05



0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0 100 200 300

Op

tica

l den

sity

at

66

0 n

m

Concentration of wild root extract

E.coli

Salmonella paratyphi

S. aureus

S. epidermidis

0

0.05

0.1

0.15

0.2

0.25

0 100 200 300

Op

tica

l den

sity

at

66

0 n

m

Concentration of cultivated root extract

E.coli

Salmonella paratyphi

S. aureus

S. epidermidis

Graph 1: Concentration of wild root extract v/s O.D. (Optical Density)

at 660 nm on different organisms

Table 2: MIC of cultivated root extract on different bacteria

Sr

no.

concentration of cultivated root

extract (µg/ml)

O.D. (Optical Density) at 660 nm on different

organism

E.coli Salmonella

paratyphi

S.

aureus

S.

epidermidis

1 0.00 0.2 0.21 0.21 0.18

2 11.20 0.18 0.2 0.19 0.17

3 56.00 0.16 0.19 0.18 0.16

4 100.80 0.15 0.18 0.17 0.15

5 168.00 0.12 0.16 0.15 0.12

6 224.00 0.07 0.1 0.11 0.1

Graph 2: Concentration of cultivated root extract v/s O.D. (Optical Density)

at 660 nm on different organisms



Table– 3: Antimicrobial susceptibility assay – wild and cultivated varieties of Eranda root.

Sr.

No.

Name of

organism

Zone of

inhibition by

wild variety

root

extract(mm)

Zone of inhibition

by cultivated

variety root

extract(mm)

Gentamycin

(concentr-

-ation 80

µg/ml)

(mm)

Fluconozocon

DT

(concentration

80 µg/ml) (mm)

150µl

(169.2

µg/ml)

200 µl

(282

µg/ml)

150 µl

(164

µg/ml)

200 µl

(224

µg/ml)

1 Escherichia coli 0 1.0 0 2.2 12 −

2 Salmonella

paratyphi

0 0 0 2.1 10 −

3 Staphylococcus

aureus

0 0 0 1.0 15 −

4 Staphylococcus

epidermidis

1.1 2.2 0 1.2 11 −

5 Aspergillus

niger

1.0 2.0 1.0 2.0 − 12



CONCLUSION

The methanol extract of (R. communis L.)

both the wild and cultivated root showed

considerable antimicrobial effect. The

cultivated variety showed inhibition towards

the growth of the gram +ve and gram –ve

organisms where as wild variety root extract

showed effect against the specific organisms S.

epidermidis and E. coli. Both the extracts

showed similar effect against the selected

fungus A. niger. The result of the study was

restricted to the selected concentration range of

the extracts. Further antimicrobial assay can be

done with a higher range of concentration.

ACKNOWLEDGEMENT

The authors wish to thank Miss Jigisha

Pancholi, Head dept. of Biochemestry, IAPS,

GAU, Jamnagar for her valuable suggestions

throughout the study. We are thankful to Mr.

Manish Vyas, Ph.D. Scholar, Dept. of RS &

BK, IPGT & RA, GAU, Jamnagar for his

support in arranging the Microbial and Fungal

strains.

REFERENCES

Abhishek M, Satish K, Sajad Y, Santosh K.

Singh, Gbks Prasad, V. K. Dua (2012).

Antimicrobial potential of Roots of

Ricinus communis against pathogenic

microorganisms, Jan-March; Issue – 1 ;

vol-2 ; 545–548.

Anonymous (1999), The Ayurvdic

Pharmacopoeia of India, edition 1st,

Govt. of India. Ministry of Health and

Family welfare, Department of I.S.M. &

H., New Delhi, Part-I, Volume –I. p.

34–35.

Capasso F, Mascolo N, Izzo AA, Gaginella TS.

(1994) Dissociation of castor oil

induced diarrhoea and intestinal

mucosal injury in rat: effect of NG-

nitro-l-arginine methyl ester. Bri J

Pharmacol; 113: 1127–30.

Ilavarasan R, Mallika M, Venkataraman S.

(2006). Anti-inflammatory and free

radical scavenging activity of Ricinus

communis root extract. J

Ethnopharmacol; 103: 478–80.

Jahir Alam Khan, Krishna Pratap Yadav

(2011), Assessment of Antifungal

property of Ricinus communis, JPBMS,

11(02).

Kirtikar KR & Basu (1985), BD. Indian

Medicinal Plants. 2nd ed, Dehradun:

International book distributor; pp.

2274–2277.

Phillips R and Martyn R. (1999) Annuals and

Biennials. London: Macmillan. P. 106.

R Yanfg LL, Yen KY, Kiso Y, Kikino H.,

(1987) Antihepatotoxic actions of

formosan plant drugs. J

Ethanopharmacology; 19: 103–10.

Saeed S and Tariq P (2007), Antimicrobial

activities of Emblica officinalis and

Coriandrum sativem against Gram-

positive bacteria and Candida albicans.

Pak. J.Bot., 39(3): 913–917.

Saeed S, Naim A and Tariq P (2007), A study

on prevalence of multi-drug- resistant

Gram-negative bacteria. Int. J. Biol.

Biotech., 4(1): 71–74.

Sandhyakumari K, Bobby RG (2003), Indira

M. Antifertility effects of Ricinus

communis Linn. on rats. Phytother Res;

17: 508–11.



Shokeen P, Anand P, Krishna Y M, Tandon V.

(2008) Antidiabetic activity of 50%

ethanolic extract of Ricinus communis

and its purified fractions. Food Chem

Toxicol; 46: 3458–66.

Shukla V.J. and Bhatt U.B. (2001), Methods of

Qualitative Testing of some Ayurvedic

Formulations, Gujarat Ayurvedic

University, Jamnagar.

Tumah H (2005); Fourth generation

cephalosporin: In-vitro activity against

nosocomial Gram-negative bacilli

compared with beta-lactam antibiotics

and ciprofloxacin. Chemotherapy, 51(2-

3): 80–85.

Visen P, Shukla B, Patnaik G, Tripathi S,

Kulshreshtha D, Srimal R, Dhawan B.

(1992) Hepatoprotective activity of

Ricinus communis leaves. Int J

Pharmacognosy; 30: 241–50.





PROBABLE ETIOPATHOGENESIS (SAMPRAPTI) OF AUTISM IN FRAME

OF AYURVEDA IN RELATION TO INTENSE WORLD THEORY

Yadav Deepmala1*

, Behera Banshidhar2, Kumar Abhimanyu

3

1Asst.Professor, Dept. of Kaumarbhritya, M.S.M. Institute of Ayurveda, Khanpur kalan, Haryana-131305,

India 2 Lecturer, Dept. of Dravyaguna, Gaur Brahman Ayurvedic College, Rohtak, Haryana – 124001, India

3 Director, All India Institute of Ayurveda, Gautampuri, Mathura road, Sarita Vihar, New Delhi-110076,

India

*Corresponding Author: E-mail: [email protected]; Mob +919414893921, +919414458895


ABSTRACT

Autism is one of the pervasive developmental disorders (PDDs). The disease affects the child's

interaction skill with the world by involving social responsiveness, communication ability and lack

of understanding for the other people showing heterogeneous clinical features of the disease. Hence,

to establish the etiopathogenesis (samprapti) of autism will help to interpret its heterogeneity and to

frame out its respective management. Hence, an effort has been made to explain the neurobiology of

autism on the basis of Ayurvedic fundamentals and „intense world theory‟ of autism. This knowledge

of etiopathogenesis may reveal the specific nature of brain dysfunction in autism and may help to

understand the development of symptomatology of the disorder and further its precise nature to

respond with various treatment strategies described in Ayurveda.

KEY WORDS: Autism, PDD, etiopathogenesis, intense world theory

Review article

Cite this article:

Yadav. D, Behera. B., Kumar. A., (2013), PROBABLE ETIOPATHOGENESIS (SAMPRAPTI) OF

AUTISM IN FRAME OF AYURVEDA IN RELATION TO INTENSE WORLD THEORY,




INTRODUCTION

According to DSM-IV, (Diagnostic and

Statistical Manual of Mental Disorders, 1994)

“Autism is a severe developmental disorder

characterized by abnormalities in social

functioning, language and communication and

unusual interests and behavior”. The key

features of autism are –

Deficient non-verbal behavior such as eye

to eye gaze, facial expression, body posture

and gestures.

Failure to develop peer relationship.

Failure to share enjoyments interests or

achievements with other people.

Impaired development of social language.

Echolalia/ idiosyncratic language.

Restricted repetitive and stereotyped

patterns of behavior, interests and activities.

Here, the process of the development of

symptomatology of autism is tried to explain

on the basis of „Intense World Theory‟

(Markram & Markram, 2010) of autism and

fundamental principles of Ayurveda

contributing in disease process.

AIMS AND OBJECTIVES

To validate the contribution of components

of Samprapti (etiopathogenesis) (i.e.

khavaigunya, tridoshas, trigunas etc.) in

pathophysiology of autism.

To establish the etiopathogenesis of autism

in frame of Ayurveda to understand the

development of symptomatology so as to

avail the multimodal treatment approaches

of Ayurveda in management of autism.

REVIEW OF CONCEPT

According to fundamentals of Ayurveda,

Samprapti (Etiopathogenesis) of any disease

comprises the vitiated dosha-dushya

sammurchhana (union). These vitiated

components are manifested in full flown

disease, when they merge with already existing

„khavaigunya‟(Shastri Kaviraj Ambikadutta,

1995) (i.e. structural and functional alteration

of body tissues/ system etc.). An etiopathology

of autism seems to be evolved with

contribution of khavaigunya in greater extent

(Flowchart- Step2). Few etiological factors

may be consequent as khavaigunya in

following way-

i. Genetic Factors: Various research studies

establish the direct link between genes and

their consequence into autism. Some of

them are briefly mentioned here (Flowchart

-step1)

Over expression of NMDA receptor gene,

particularly the receptor subunits NR2A

and NR2B as well as the CAM Kinase

linked second messenger pathway are

observed in autistic brains. (Rinaldi et al.,

2007).

Several studies indicate the involvement of

glutamatergic systems in autism. Single

nucleotide polymorphism (SNPs) in the

gene encoding "glutamate6 receptor"

(GLUR6 -Jamain et al., 2002) and

“glutamate8 receptor” (GLUR8 - Serojee et

al., 2003) were reported in autistic brains.

Also, several glutamatergic synapse gene

mutations on chromosome 22 were also

associated with autism (Jamain et al.,

2003).

Rett‟s Syndrome (RTT), a trait of autism, is

an X-linked dominant progressive

neurodevelopmental disorder which

exhibits all the three core characteristics of

autism. It is caused by mutations in the

gene encoding methyl CPG-binding protein

(MeCP2). (Amir et al., 1999)

Similarly fragile X-syndrome is another X-

linked disorder which displays the features

similar to autism such as abnormal speech

pattern, stereotypic movements and

abnormal social behavior, particularly

shyness and limited eye contact. It is caused

by the mutations in the FMR1 gene that

encodes for fragile X-mental retardation

protein (FMRP).



Flow chart 1: Probable Etiopathogenesis of Autism



ii. Epigenetic toxic insults: (Flowchart-

step2)

A recent study of autism provides a link

between hyper-activation of NMDA

receptor and deficiency of hypothalamic

inhibitory hormone i.e. digoxin. Lack of

digoxin may fail to avoid brain damage due

to excitotoxicity (Kurup & Kurup, 2003)

and consequent in autism due to toxicity

reasoning.

Imbalance of inhibition-excitation activities

of neuronal pathways due to altered levels

of serotonergic, GABAergic and NMDA

neurotransmitters level may result in

epigenetic toxic insults.

iii. Postnatal environmental factors:

(Flowchart-step3)

Several perinatal environmental factors

may cause autism. These factors include

obstetrical complications, prematurity,

hypoxic-ischemic encephalitis, jaundice etc.

MMR Vaccine and certain drug toxicity are

causally linked to the development of Autism.

Thus, above three etiological factors are

responsible for the structural and functional

alterations in brain (Khavaigunya). This will

further activates the molecular imbalance in

cellular and circuit level that sensitizes gene

expression pathways to respond excessively to

environmental stimulation. (Flowchart-step4)

Under normal condition, gene expression

pathways would enable for enriched

environments to nurture brain development but

if these pathways are sensitized then

environmental stimulation may cause

exaggerated and accelerated development of

brain. This will further affect the neuron

connectivity in autistic brain. Several MRI

studies suggest that in autistic brain long-range

connections (essential for complex information

processing of higher order functions) between

different brain areas are underdeveloped while

the short-range connections (essential for

primary information processing) are overly

developed. (Just et al., 2007; Mottran et al.,

2006)

According to Ayurvedic perspective to

maintain proper connection between body

tissues is a function of pranavayu (i.e.

sandhankar karma) (Chaturvedi G. & Shastry

K., 1996). Therefore, the hyper-connectivity

between short-range synapses and under-

connectivity between long-range synapses may

be caused by vitiated pranavayu. Similarly to

maintain the intactness between these

connections or of body tissues is a function of

kapha dosha (i.e.sneha, bandha & sthiratva)

(Chaturvedi G. & Shastry K., 1996). But the

impaired kapha dosha may lack to provide

nourishment (Snehana) to brain matter and

affects its compactness and stability which

ultimately makes brain tissues more vulnerable

to endogenous cytotoxicity.

Thus, these alterations in neural

connectivity leads to cascade of events which

comprises the symptomatology of autism in a

following way-

A. Hyper-connectivity to short-range

primary sensory pathway

Due to hyper-connectivity between short

range neurons, the flow of primary sensory

information speedily transfers via synapses

causing hyper reactivity across different brain

regions (Flowchart-step 4, 5, 6). In Ayurvedic

context, this process is carried out by enhanced

„chalatva guna‟ (quality) of vata dosha

(Chaturvedi G. & Shastry K., 1996).This

hyper-reactivity contributes central role in

pathophysiology of autism, which can be

interpreted as below–

i. Locus Coeruleus (LC):

Super-charged micro circuits in primary

sensory area may produce enhanced sensitivity

to sensory stimulation. This may consequently

over sensitize to locus coeruleus for upcoming

environmental stimulation and may result into

enhanced nor epinephrine secretion, which

further leads to over excitation of NE pathway

(Flowchart-step7).

Over excitatory NE pathway is cause for

hyper-perception and hyper-attention. As per a

scientific study, higher sustained attention is

reported in autism as compare to control group



i.e. autistic people have the ability to maintain

attention to repetitive stimuli over prolonged

periods of time (Johnson et al., 2007). This

may hamper the capacity of selective attention

for relevant sensory stimuli and thereby

sensory overload. (Flowchart-step8)

In an Ayurvedic context, vitiated pranavayu

and kapha dosha may lead to impairment of

function of manas (Chaturvedi G. & Shastry

K., 1996) and the function of dhee

(Chaturvedi G. & Shastry K., 1996) which

result into abnormal orientation and

engagement of all relevant and irrelevant

environmental stimulation, in turn causes

for sensory overload.

ii. Neocortex and Amygdala:

In pathophysiology of autism, neocortex

particularly prefrontal and somatosensory

cortex and amygdala are involved in great

extent. At neocortical region, the hyper-

excitatory NE pathway are inhibited by

increased GABAergic pathway indicating that

inhibition may able to recruit a constant

matching level of excitation without developing

an imbalance (Rinaldi et al., 2008). This will

confront the neocortex for excessive processing

of primary sensory information. (Flowchart-

step9)

This can be illustrated by an example of

sensory perceptions. Primary processing of

environmental stimuli of sensory origin (i.e.

visual, auditory, touch etc.) may be carried out

excessively by somatosensory cortex while

cognitive functions related to simple feature are

carried out excessively by prefrontal cortex.

Similarly, Amygdala may also become overly

reactive for processing emotionally relevant

information. It is widely established that the

amygdala mediates the formation and storage

of fear memories (Le Daux, 2003) and

enhances memory formation throughout the

other brain regions by acting as an emotional

amplifier (Cahill & Mc Gaugh, 1996). Hence in

autistic people, dysfunction of amygdala may

result into exaggerated and more persistent

processing and storage of aversive emotional

and fear related memories. This is supported by

the theory of imbalance of excitation inhibition

which would result into increased reactivity,

due to loss of inhibition at amygdala (Casonova

et al., 2003).

Thus, the hyper reactive neocortex and

amygdala may significantly consequences into

excessive perception, attention, learning and

emotionality (Flowchart-step10). This process

of hyper learning is stored in the form of simple

features of touch, sound, light, fear, emotions,

language etc. (Flowchart-step11).

Hippocampus, Basal ganglia and amygdala

are the sites, where memories of these simple

features are allocated (Flowchart-step12). Long

term potentiation (LTP) is the neuronal

mechanism which is widely assumed to

underlie memory formation. This LTP

mechanism is mediated by glutamatergic

neurotransmitters and receptors system

particularly NMDA (Nicoll & Malenka, 1999)

and alterations in this system may contribute to

the above observed hyper-plasticity leading to

hyper-memory and hyper-learning. (Flowchart-

step13)

With excessive learning and memory

processes, sensory regions may consolidate into

overspecialized modules and lead to hyper-

preference processing pathway. (Flowchart-

step14)

During early development (probably before

age 3 years), this may lead to excessive flow of

information from sensory areas to the higher

integration areas such as association cortices

and prefrontal lobe which may cause

prematurely accelerated growth of these higher

order brain areas as truly observed in autism.

This would be the reason of why autistic

children have certain unusual talents and older

autistics are excellent on task involving long

term memory like recall of train time-tables,

historical dates, chemical equations or recall of

the exact words of songs heard years before

(Carper et al., 2002)

Ultimately the hyper-preference processing

in the sensory domain, may lead to exaggerated

selectivity, sensitivity and specialization of

simple sensory features. As a consequence-



Autistic children would remain with

fragmented and amplified perception of bits

and pieces of the world.

Autistics may have abnormal and obsessive

attention to detail and hence can notice the

smallest change in their environment.

Autistics may become hyper-focused in

arbitrary subjects of interests and sustain

their attention on these subjects for

unusually long time periods.

On the whole, this hyper-connectivity

phenomenon may cause for enhanced

perception of sensory fragments; focus on

details and deficit in complex and more holistic

processing.

As per the Ayurvedic aspect, pranavayu

promotes and regulates the other

biophysical components and sense faculties

(by the virtue of prerana karma) to perform

their respective functions (Chaturvedi G. &

Shastry K., 1996). But as described earlier,

vitiated prana vayu may unable to perform

its function of synaptic connectivity

properly and so as to fail to regulate the

functions of other components, rather

consequently responsible for impairment of

manas, buddhi and other doshas.

(Flowchart-step7,8)

Due to impaired manas, the functions

related to motor control, abstract thinking

and thoughtful planning may upset and lead

to non-oriented information processing

(Chaturvedi G. & Shastry K., 1996)

(Flowchart-step9).

Due to impaired buddhi, particularly dhriti,

the function of selective attention and

further retention may disturb leading to

excessive storage of primary information

(Chaturvedi G. & Shastry K., 1996)

(Flowchart-step10).

Functions of sadhak pitta simply resemble

with the functions of neocortex and

amygdala. As cognitive functions

represented by sadhak pitta can be

represented in the form of buddhi and

medha

(Arunadutta, 2002) as well as

function of emotional & social cues are

represented as bhaya, shaurya (Chaturvedi

G. & Shastry K., 1996) etc. But due to

derangement of sadhak pitta, the

exaggerated processing of sensory,

cognitive and emotional cues take place

thereby leading to overflow of primary

knowledge (Flowchart-step11, 12).

Memorization i.e. smriti is a function

carried out by udan vayu (Arunadutta,

2002). But due to its derangement and

hyper preferential mnemonic pathway,

autistic child may show excessive retrieval

of working memory as well as long term

memory of only primary features

(Flowchart-step13). But in older autistics,

working memory functions are seen widely

upset, although long term memory may

remain intact due to repetitive maintenance

rehearsal. This may be suggestive of

ongoing degradation of Udan vayu due to

local background pathology (Flowchart-

step14).

Concept of hyper-memory can also be

explained with the help of an Ayurvedic

principle i.e. „Samanyam Vriddhikaranam‟

(Chaturvedi G. & Shastry K., 1996).

According to this theory, generic

concomitance may augment the same class

of characters, in turn may responsible for

their overloading.

Such an overspecialized hyper-memory

may tend to activate even by the mild

environmental stimuli of same class of

knowledge and further integrate with its

processing to develop hyper-preferential

pathway.This mechanism is also supported

by one of smriti-hetu (Factors responsible

for memory) i.e. „sadrishyat

smritirupajayate‟ (i.e. similarity between

current knowledge and previous

experience). (Chaturvedi G. & Shastry K.,

1996) (Flowchart -step14).

Back on continuing modern

pathophysiology of autism, in general, it is

observed that the autistic children may seem

normal, rather, gifted at the initial period of

development, the symptomatology of autism



initiates after the age of 18 months or before

the age of 3 years when child begins to learn

more complex task. The symptomatology may

be driven by under-connectivity of long range

of connections required for complex task.

B. Under-connectivity to long range

complex sensory pathway

As described earlier, long range neuronal

connections play a key role in complex

information processing (Minshew & Goldstein,

1993). But in autistic brain, under-connectivity

may observed between long range columns of

different brain regions. Hence there is an

increasing impairment in integrating

progressively more complex information across

different brain regions which in turn results into

memory deficits for complex and abstract

material (Flowchart- step15).

Also the background mechanism of hyper-

preferential sensory-mnemonic pathway of

short-range columns, also inhibit the higher

order information processing. As on exposure

to environmental stimuli for complex task

(Flowchart -step16), the hyper-preferential

pathway become activate and enhances its

sensitivity for selective simple features. This

may synergistically processed by the

consolidated mnemonic inputs (Hyper-

memory) of previous primary information and

results into dominance of the earliest features

and avoidance of processing of other features.

Such hyper-autonomous and overly selective

pathway leads to following symptoms with

respect to different brain regions (Flowchart -

step17).

i. Somatosensory cortex : (Flowchart-

step18)

Under-connectivity to long range circuits

and hyper-connectivity with microcircuits in

neocortex, particularly somatosensory cortex

causes exaggerated perception and attention of

fragments of sensory world which must be

holistically processed at normal circumstances.

This may enhance by hyper plasticity

component which drives exaggerated memories

to further amplify the processing of same

stimulus and also drive over generalization of

attention to all related forms of the stimulus.

Thereby the positive consequences are

exceptional capabilities for primary and

specific tasks while the negative consequences

are impairment of holistic processing and a

limited repertoire of behavioral routines, which

may further repeated obsessively i.e.

stereotyped behavior, a core characteristic of

autism (Flowchart -step19).

Similarly in the domains of auditory, visual

& touch stimuli, autistic people on positive

consequence may exhibit enhanced

discrimination capabilities for elementary

stimuli and on negative consequence, they may

exhibit the diminished global interference for

complex stimuli due to impaired pre-pulse

inhibition (Foxton et al., 2003 Mottorn et al.,

2003). This will lead to hypersensitivity to

environmental sound, light and touch stimuli,

which is observes as key characteristics of

autism (Flowchart -step 20).

In an Ayurvedic paradigm, stereotyped

behavior and hyper-sensitivity to touch,

light and sound are the features

demonstrated by impaired vyan vayu

(Arunadutta, 2002) (Flowchart -step19, 20).

ii. Prefrontal Cortex: (Flowchart -step21)

Downside of hyper-connectivity and under

connectivity of cognitive regions, particularly

PFC, indicates impairment in higher executive

functions. Excessive memory in low level

sensory and elementary cognitive regions may

lead to an early over-specialization of primary

feature processing, missed developmental

opportunities to acquire a full spectrum of

primary processing strategies and to build

higher order strategies. This might lead to a

fragmented alphabet of feature processing

capabilities in the vocabulary of sensory

processing and obstruct the development of

higher cognitive functions such as abstract

thinking and language processing. Thus, this

mechanism is responsible for language and

speech impairment, which is a characteristic of

autism (Flowchart -step22).

In an Ayurvedic paradigm, derangement of

udan vayu is responsible for speech

impairment (Arunadutta, 2002).



iii. Amygdala:

In autism the functional impairment of

amygdala is resulted into storage of aversive

emotional and fear related memories

(Flowchart- step23). Later on, exposure to fear

stimuli, this preformed memories may lead to

progressive generalization of fear which may

consequence into behavior and may account for

inappropriate reactions to the environment,

sudden & inexplicable anxiety attacks, loss of

fitness required for social interaction and

phobias. These are also core characteristics of

autism. (Flowchart-step24)

Impairment in social interactions &

communication are other characteristics of

autism, i.e. which are also mediated by

insufficient activation of amygdala (Pierce et

al., 2001). Due to hypo-activation of amygdala,

autistic people may become severely unable to

“read other people minds” (Flowchart-step25)

and empathizing with other people by affecting

the following two elements-

Ability to distinguish between oneself and

others and realize that other people have

independent minds and may pursue

different goals from one-self.

The ability to express an appropriate

emotional reaction to the other person‟s

mental state, thus to be unable to empathize

with others mind.

On the whole, the deficits in language &

speech, undue fear & social phobia, and

inability to read other people's mind may

altogether mediate impairment in social

interaction and communication. (Flowchart-

step26)

In Ayurvedic paradigms the features of

emotionality is connoted by Sadhak pitta

(Chaturvedi G. & Shastry K., 1996) i.e.

bhaya harsha, prasad, krodha, etc. Thus

vitiated sadhak pitta may responsible for

increased fear and anxiety which lead to

social withdrawal and avoidance.

iv. Fusiform face area (FFA):

Eye contact and watching the facial expressions

are one of the first signs of cognitively healthy

infants and serve to build the basis for

successful navigation through a social

environment. But deviant eye gaze is a core

characteristic observed in autistic child. This is

driven by the impaired cortical region, named

as fusiform face area. As suggested by its

name, in normal subjects this area is highly

reactive to face recognition. But in autistic

children, fusiform face area is observed to be

hypo-reactive which lead to abnormal face

perception and social avoidance. (Kanwishar et

al., 2000) (Flowchart-step27)

Due to hypo-reactivity of FFA, amygdala

may have to confront to response the face and

eye recognition stimuli. On exposure, the right

amygdala shows greater activation when

viewing familiar and unfamiliar faces while left

amygdala and left orbito-frontal cortex shows

greater activation on viewing emotional faces.

Both areas forms part of the emotion circuit of

the brain and shows heightened emotional

response to these stimuli. Amygdala,

simultaneously also makes quick and powerful

fear associations with fearful mnemonic-inputs.

In consequence to this, Autistic child may

spend less time fixating with the eye region

resulting into avoidance of eye contact which is

a characteristic of autism (Dalton et al., 2005)

(Flowchart -Step28 )

In an Ayurvedic paradigm, eye movement

is a function of vyan vayu. So deviant eye

gaze may also the result of deranged vyan

vayu (Arunadutta, 2002).



Table 1: Probable etiopathological markers of Autism as per Modern in relation to Ayurveda

Modern Ayurveda Reference

1.

Genetic factor, epigenetic toxic

insults, post natal environmental

factor

Khavaigunya Shastri Kaviraj

Ambikadutta ,1995

2. Dysregulation of gene expression

path way Khavaigunya

Shastri Kaviraj

Ambikadutta ,1995

3.

Hyperconnectivity to short-rang

pathway of primary features Impairment of

sandhankar karma of

pranavayu

Chaturvedi G. & Shastry

K., 1996 Underconnectivity to long-range

neuronal pathway

4. Information speedly transfer via

synapses

Impairment of chalatva

quality of Vata dosha


K., 1996

5. Increased connectivity of NE

pathway

Impairment of

pranavayu


K., 1996

6. Hyperperception and hyperattention Impairment of Dhee Chaturvedi G. & Shastry

K., 1996

7. Somatosensory cortex, PFC &

amygdala

Function similar to

manas and buddhi


K., 1996

8. Excessive storage of all information

Based on principle of

“ samanyam vriddhi

karanam”


K., 1996

9. Hyper memory Increased Udanvayu Arunadutta, 2002

10. Hyper preferential pathway Excessive intake of

smritihetu


K., 1996

11.

Stereotyped repetitive behavior

Impairment of Vyanvayu Arunadutta, 2002 Hyper sensitivity to light, touch,

sound

12. Impairment of language Impairment of Udan vayu Arunadutta, 2002

13. Fear, anxiety and phobia Impairment of Sadhak

pitta


K., 1996

14. Lack of communication and social

Impairment

Impairment of Udan vayu

and Sadhak pitta

Arunadutta, 2002,


K., 1996

15. Lack of eye communication Impairment of Vyana

vayu Arunadutta, 2002

CONCLUSION

Etiopathogenesis of autism may

predominantly evolve from khavaigunya as

consequence of various sahaja (genetic) &

agantuja (Epigenetic toxic insults & post natal

environmental factor) hetus. Vitiated doshas

(physical & mental) may exacerbate the

khavaigunya leading to various core features of

autism. Hyperconnectivity & underconnectivity

among short range & long range neuronal

pathways respectively may evolve from

impairment of pranavayu which in turn causes

for hyper-perception, hyper-attention & hyper-

memory (i.e. impairment of functions of manas

& buddhi). Impairment of vyana vayu may



responsible for stereotyped repetitive behavior,

hypersensitivity to light, touch & sound and

lack of eye communication. Deranged

Udanvayu may result into echolalia and

language impairment while deranged Sadhak

pitta may cause fear, anxiety and phobia. These

features, ultimatly contribute to lack of

communication & social impairment. On the

whole, this Ayurvedic framing of

etiopathogenesis of autism may help to

understand the contribution of pathological

markers in developing symptoms of disease.

This knowledge can be further used in

Ayurvedic management of autism by reducing

the effect of „khavaigunya‟ through restoring

the functions of other pathological markers and

normalizing the functions of manas, buddhi

(with dhee, dhriti and smriti), vitiated doshas

and trigunas, which in turn, may helpful in

reducing problematic behavior that would be

adaptive at lower rates as well as maintaining

this adaptive behavior for longer duration. In

addition, it may help in acquiring new skills of

leaning and communicating.

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CRITICAL APPRAISAL SKILL FOR THE ARTICLE PUBLISHED IN

AYURVEDIC HEALTH DOMAIN

Vaikos C D1*, Waghchoure Ashok

2

1Associate professor and Head of Department, Dept of Rachana Sharir, Govt. Ayurved College, Nanded,

Maharashtra, India 2Assistant professor, Dept of Rachana Sharir, Govt Ayurved College, Nanded, Maharashtra, India

*Corresponding Author: Email - [email protected]


ABSTRACT

It is the need of all Ayurvedic students, teachers and scientists to have complete knowledge of the

process of evaluation and appraisal of research, review or original articles published in peer reviewed

valid journals. Most of the students (and more or less teachers also) are ignorant on the appraisal of a

published article since this was not part of their designed curriculum. Central council of Indian

medicine have designed curriculum of research methodology recently, which includes publication

skills. Published articles on Ayurvedic subjects must be evidence based. This article will help to

develop an innovative approach on how to teach and learn critical appraisal skills to Ayurvedic

students and hence user could decide how much he/she can rely on the results of the article and,

therefore, shall or shall not use the results of that article in his/her practice. Research studies,

including qualitative studies, form the basis for evidence-based practice among health professionals.

However, many practicing Ayurvedic health educators do not feel fully confident to appraise

qualitative and quantitative research studies. This review provides guidelines for appraising the

strengths and weaknesses of published qualitative research articles and Health educators will be

better equipped to evaluate the quality of the evidence through critical appraisals of qualitative

research publications. Published research studies, including qualitative studies, provide the evidence

for the selection of evidence-based practices in health education and promotion. For health educators,

critical reading of research studies can be time consuming and challenging, however they can

increase their confidence in appraising research studies by using the guidelines described in this

article.

KEY WORDS: Evaluation, appraisal, central council of Indian medicine, evidence based medicine,

quantitative and qualitative study, Ayurveda

Review article

Cite this article:

Vaikos C. D., Waghchoure. A. (2013), CRITICAL APPRAISAL SKILL FOR THE ARTICLE

PUBLISHED IN AYURVEDIC HEALTH DOMAIN,


mailto:Email%20-%[email protected]



INTRODUCTION

Critical appraisal has been defined as The

application of rules of evidence to a study to

assess the validity of the published research

data (wikipaedia.org), completeness of

reporting, methods and procedures,

conclusions, compliance with ethical

standards,” etc. Critical appraisal is the skill

that the science of epidemiology offers to

scientists and specialists in different disciplines

including medicine (Timm DF, Banks DE,

McLarty et al., 2012, McAllister D, Wild S et

al., 2009, Green BN, Johnson CD, et al., 2007).

It is used to evaluate and determine the positive

and negative aspects of a published article

(Mohsen Rezaeian et al., 2006), in evidence-

based healthcare training, to assist in clinical

study in decision-making and in evidence-

based social care and education provision

(wikipaedia.org).

Due to the rapid - development, spread and

worldwide acceptance of Ayurveda, the expert

specialities, students, clinicians, health

professionals’, teachers, and research scientists

in Ayurvedic field needs to develop efficient

skills in critical appraisal of published articles

to use or not to use results of that article in

support of his/her articles. Ayurveda is an

evidence based science, selection of relevant

literature, understanding of research findings,

elicit patients own preferences etc (Jane M

Young, Michael J Solomon, 2009) and use of

journal club and letter-to-the-editor writing

project (Green BN, Johnson CD, 2007) may

create ability in clinician to appraise the

research articles. Journal club should be a part

of curriculum which will provide exposure to

Ayurvedic student to the world of frontline

research and safe way to enter in the world of

translational research (Tamoghna Biswas,

2011). The quantity of published articles in the

databases like Medline, Pubmed, Medknow etc

is increasing. According to Medline database

more than 12,000 new articles, including

papers on nearly 300 randomized controlled

trials (RCTs), are added to their database in

each week (Glasziou PP et al., 2008) but the

question regarding the validity of results and

their use in local help is merely doubtful.

Therefore every article should be critically

scrutinised to evaluate the robustness of the

findings. Many times the articles on

quantitative and qualitative research are

questioned by scientific scholars that they lack

scientific rigour, credibility, reliability,

trustworthiness and validity. In such cases it is

necessary to audit the process up to the end

product. This will help the user/reader to

determine whether the claim made by the

researcher is justifiable or not (Dorothy

Horsbugh, et al., 2003).

Qualitative research studies are particularly

helpful when looking at context, experiences or

meanings related to principals/laws of

diagnostics/therapeutic measures or an issue;

(for e. g. theory of Panchmahabhootas (five

basic constituent element), Prakriti (body

constitution), Samanya Vishesha Sidhdhanta,

Lokpurusha Sidhdhanta etc). Quantitative

research is useful to address the intervention or

therapy questions (Jeanfreau, S. & Jack, L.

2010) or drug discovery or safety and toxicity

study of herbo-mineral or mineral drugs. Both

kinds of research, evidence may be necessary

for a complete understanding of how and why

an issue occurs (qualitative research questions)

and what can be done to address the issue

(quantitative research).

DISCUSSION

Articles on various subjects of Ayurved -

viz. standardization of Ayurvedic drugs,

clinical trial of mineral, herbal and herbo-

mineral drugs mentioned in Ayurvedic texts,

conceptual/empirical research on the basic

principle like Tridosha Sidhanta, (Theory of

three humours of body) Prakriti (theory of

body constituent) etc are published in various

journals. There are no certain or specific

criteria in Ayurvedic texts to assess the strength

and weakness of articles published in various

national and international peer reviewed

journal. Hence criterion based on modern

science is considered for the appraisal of

articles in Ayurvedic subject. Following

discussion with some modification according to

the research question may help the Ayurvedic



research scholar to appraise their own and

others articles before publishing.

Appraisal scheme (strength and weakness of

published qualified articles)

How do we appraise? Appraisal can be

done using common sense without the help of a

statistical expert (Sarah Lawson et al., 2010).

Validity and relevancy of various articles can

be considered using different types of checklist.

Anyone can assess and evaluate the quality

of article using questionary of 10 questions in

two sets, one is primary question set and other

is main question set which will consider – the

results of research study, validity of review

study and its help to everyone or local one?

(Joan Durrance et al., 2009). These question

sets are more or less applicable to evaluate the

interventional and observational study.

Screening question/Primary question

1. Aims of the research is mentioned by author

or not? What Type of Research Question Does

the Study Pose? (Jane M Young, Michael J

Solomon, et al., 2009).

Consider: the goal of the research, its

important and its relevance.

2. Adoption of appropriate methodology.

Consider: If the research seeks to interpret

or illuminate the actions and/or subjective

experiences of research participants.

Detailed/Main questions

3. Research design is appropriate to address the

aims of the research and is the study's research

question relevant (Jane M Young, Michael J

Solomon et al., 2009).

Consider: If the researcher has justified the

research design (e.g. have they discussed how

they decided which method to use)?

4. Whether the recruitment strategy (as regards

to sample size, sample design, inclusion and

exclusion criteria etc.) was appropriate to the

aims of the research and did the study methods

address and eliminates the key potential

sources of bias? (Jane M Young, Michael J

Solomon et al., 2009)

Consider: if the researcher has explained

how and why the participants were selected. If

there are any discussions around recruitment

(e.g. why some people chose not to take part).

5. Were the data collected in a way that

addressed the research issue? Was the Study

Performed in Line with the Original Protocol?

(Jane M Young, Michael J Solomon et al.,

2009)

Consider: If the setting for data collection

was justified. If it is clear how data were

collected (e.g. focus group, semi-structured

interview etc.).If the researcher has justified the

methods chosen. If the researcher has made the

methods explicit (e.g. for interview method, is

there an indication of how interviews were

conducted, or did they use a topic guide)? If

methods were modified during the study. If so,

has the researcher explained how and why? If

the form of data is clear (e.g. tape recordings,

video material, notes etc.) If the researcher has

discussed saturation of data.

6. Has the relationship between researcher and

participants been adequately considered?

Consider: If the researcher critically

examined their own role, potential bias and

influence during: 1) Formulation of the

research questions 2) Data collection, including

sample recruitment and choice of location.

How the researcher responded to events during

the study and whether they considered the

implications of any changes in the research

design.

7. Have ethical issues been taken into

consideration?

Consider: If the research was explained in

sufficient details to participants. If the

researcher has discussed issues raised by the

study (e.g. issues about informed consent,

confidentiality and how they have handled the

effects of the study on the participants during



and after the study). If approval has been

sought from the ethics committee.

8 Was the data analysis and statistical analysis

is sufficiently rigorous and correct? Does the

Study Test a Stated Hypothesis? (Jane M

Young, Michael J Solomon et al., 2009)

Consider: if there is an in-depth description

of the analysis process. If thematic analysis is

used. If so, is it clear how the categories/themes

were derived from the data? Whether the

researcher explains how the data presented

were selected from the original sample to

demonstrate the analysis process. If sufficient

data are presented to support the findings. To

what extent contradictory data are taken into

account. Whether the researcher critically

examined their own role, potential bias and

influence during analysis and selection of data

for presentation.

9. Do the Data Justify the Conclusions? Is there

a clear statement of findings?

Consider: If the findings are explicit. If

there is adequate discussion of the evidence (in

favour and against the researcher’s arguments).

If the researcher has discussed the credibility of

their findings (e.g. triangulation, respondent

validation, more than one analyst). If the

findings are discussed in relation to the original

research question.

10. How valuable is the research? Does it add

anything new? (Jane M Young, Michael J

Solomon et al., 2009)

Consider: usefulness of research, if the

researcher discusses the contribution the study

makes to existing knowledge or understanding

e.g. do they consider the findings in relation to

current practice or policy, or relevant research-

based literature? If they identify new areas

where research is necessary. If the researchers

have discussed whether or how the findings can

be transferred to other populations or

considered other ways the research may be

used (Joan Durrance, 2009).

Software for appraisals

There are software tool which helps to

create Critically Appraised Topics, or CATs,

(CEBM oxford university) for the key articles

you encounter about Therapy, Diagnosis,

Prognosis, Aetiology/Harm and Systematic

Reviews of Therapy (McAllister D, Wild S et

al., 2009).

This software may help the

Ayurvedic researcher to appraise his or others

article before publishing. Though in Ayurvedic

fraternity, such software is not available but

one can use it by making some modification

with the help of software experts based on the

concept of Ayurvedic science.

CONCLUSION

The information in this article can provide

deeper understanding and appreciation for

published qualitative research. The appraisals

of published article using guidelines, questions,

and explanations provided in this article are not

intended to be all inclusive and may require

some modification to answer and assess the

research question. No study is perfect, nor does

any study answer all questions, It is

recommended that qualitative study and its

value of evidence must read and assessed

critically. It is concluded that rigorous criteria

is available to evaluate the quantitative and

qualitative research (Popay J., Rogers A. &

Williams G. et al., 1998). Software is also

available to assist the researcher to evaluate the

strength and weakness of their own and

published article. This software is designed for

modern research which requires some

modifications to meet the need of research

question. The use of quantitative criteria to

evaluate qualitative research may create the

impression that the latter is not academically

rigorous. Hence the evaluation criteria for

qualitative and quantitative research should be

different.



REFERENCES

CAT (critical appraisal tool) maker, centre for

evidence based medicine, oxford

university)

http://www.cebm.net/?o=1216

Dorothy Horsbugh, (2003) Evaluation of

qualitative research, Journal of Clinical

Nursing 12: 307–312

Glasziou PP (2008), Information overloads:

what's behind it, what's beyond it? Med

J Aust 189: 84–85

Green BN, Johnson CD (2007) use of a

modified journal club and letters to

editors to teach critical appraisal skills.

J Allied Health; 36:47–51

Jane M Young, Michael J Solomon, (2009)

How to Critically Appraise an Article,

Nat Clin Pract Gastroenterol

Hepatol;6(2):82–91.

Jeanfreau, S. & Jack, L. (2010) Qualitative

research appraisal tool

http://www.nccmt.ca/registry/view/eng/

148.html

Joan Durrance (2009) Creative appraisals skill

program

http://creativecommons.org/licenses/by-

nc-sa/3.0/

McAllister D, Wild S. (2009) Why should

clinicians understand epidemiology?

Post Grad Med J 85:31, Med J Aust

189: 84–85

Mohsen Rezaeian, Department of Social

Medicine, Rafsanjan Medical School,

Rafsanjan University of Medical

Sciences,Rafsanjan, How to teach

medical students to critically appraise a

published article in the public health

domain Iran, http://www.jehp.net

Popay J., Rogers A. & Williams G. (1998)

Rationale and standards for the

systematic review of qualitative

literature in health services research.

Qualitative Health Research 8(3), 341–

351.

Sarah Lawson, (2010) Critical appraisal of

qualitative research

Tamoghna Biswas, (2011) Indian Role of

journal clubs in undergraduate medical

education, Journal of Community

Medicine vol 36

Timm DF, Banks DE, McLarty J. (2012),

Critical appraisal process: Step-by-step.

South Med J; 105: 144–8.

Wikipedia.org

http://en.wikipedia.org/wiki/Critical_ap

praisal


http://www.ijcm.org.in/searchresult.asp?search=&author=Tamoghna+Biswas&journal=Y&but_search=Search&entries=10&pg=1&s=0




TRADITIONAL USE OF KAHU (LACTUCA SCARIOLA L.) - A REVIEW

Arif Mohammad1*

1HSZH Government Unani Medical College & Hospital (Barkatulla University), AYUSH Campus, Nehru

Nagar Kolar Bypass Road, Bhopal-462003

*Corresponding Author: E mail: [email protected]


ABSTRACT

Lactuca scariola Linn (prickly lettuce) is an important drug in Unani system of medicine.

Tukhme kahu (Prickly lettuce seed) has traditionally been used for the treatment of headache,

insomnia, nervousness, hypertension, palpitation, fever etc. Recently the discovery of active

components from the plant and their biological function in disease control has led to active interest in

the plant across the globe. It is easily available and cost effective drug which has drawn interest of

many researchers and is screened for various bioactive substances. The present article explores the

Unani classical and published scientific literature to compile the traditional and scientific data

comprising pharmacognostic description, pharmacological studies, therapeutic uses, and safety

profile of Lactuca scariola to date. The drug exhibits varied pharmacological activities such as

sedative, hypnotic, diuretic, deobstruent, antipyretic, anti inflammatory, blood purifier, demulcent,

refrigerant, anesthetic and antispasmodic, anti cancer, antibacterial, bronchodilator and vasorelaxant.

The present review will provide comprehensive information on phytochemical and therapeutic uses

with special reference to Unani medicine which will help to tap its unexplored potential with more

scientific approach.

KEY WORDS: Lactuca scariola, Tukhme Kahu, Unani medicine, Sedative.

Review article

Cite this article:

Arif Mohammad (2013), TRADITIONAL USE OF KAHU (LACTUCA SCARIOLA) - A REVIEW,





INTRODUCTION:

Tukhme Kahu is the seed of Lactuca

scariola Linn (Family - Asteraceae) which is

commonly known as “Prickly Lettuce” seeds or

“Wild Lettuce” seeds. L. virosa is a variety

closely related to L. scariola. The species

Lactuca sativa is the common or garden variety

which is cultivated in many parts of India as a

culinary vegetable (Nadkarni, K.M. 1954). The

generic name Lactuca and the common name

Lettuce derived from the Latin world lactus

(milk), a milky fluid that flows from the stems

when they break or are cut (Murray, J.A. 1983,

Bunney, S.1992). It has been described by

different closely related botanical names such

as Lactuca sativa, L. capitata, L. virosa, L.

bracteata and sativa wall. L. capitata, L. virosa

(Nadkarni, K.M. 1954, Anonymous 1962,

Watt, G. 1972, Khorey, R.N. et al.1985). Kahu

plant and Tukhme Kahu has been used for a

long time. According to Decandolle, 1895, it

has been cultivated for more than 2000 years.

Its medicinal properties were described by

Hippocrates (430 BC). It was praised by

Aristotle (356 BC). The species were described

by Theophrastus (322 BC) and Dioscorides (60

AD). Galen (164 AD) gave the idea of general

use. Arab physicians like Altabri (d. 861 CE) in

“Firdausul Hikmat”, Razi (865-925 CE) in

„Alhawi‟, Ibnesina (980-1031 CE) in Alqanoon

and Majusi (d. 1028) in “Kamil-us-Sana” have

described the pharmacological actions and uses

of Tukhme Kahu in detail under the heading of

Khas and Bazrul Khas (Kahu seeds). The

common reported pharmacological actions are

anxolytic, sedative, antipyretic, diuretic and

analgesic which have been identified and

proved by scientific studies. Apart from the

reported pharmacological actions, Lactuca

scariola also have Anti cancer, antibacterial,

Spasmolytic, bronchodilator and vaso-relaxant

Activities. Keeping in view the increase

incidence of cancer, increase resistance of

antibiotics and failure of management of

hypertension & asthma which are the leading

problem of the healthcare system, the present

review was conducted which aimed to provide

the current and ancient literary knowledge to

the researchers of the related fields.

Unani classical literature and ethno medical

literature on recent developments in research

on Tukhme Kahu (Lactuca scariola) including

original articles and papers were taken into

study for the report. All the reports of

phytochemical, pharmacological and clinical

studies from animal and human model system

were included in the review. Reported data was

analyzed and represented in the form of table

for the current review.

Vernaculars

Tukhme Kahu is popularly known in

different dialect and languages as follows.

Bazrul Khas (Arabic) Kahu, Salad Beej

(Bengali) Prickly Lettuce, Wild Lettuce

(English), Thridox (Greek), Guado, Lattuga

(Italian), Kahu (Punjabi), Sallattu Virai

(Tamil), Kavu Vitula (Telugu), Kahu ke beej

(Hindi), In Persian and Urdu, the drug is

commonly known as Tukhme Kahu

(Nadkarni, K.M. 1954).

Morphology

Ibne Baitar (1197–1248 H) described in

“Aljame-ul-Mufradat-ul-Advia wa Aghzia”

with reference to Dioscorides (60 AD), Galen

(121–200 AD), Ibne Masoya and Razi (865–

925 AD) as follows:–

Kahu (Khas) actually has two varieties based

on its occurrence.

1. Bustaani (Baaghi) “Lactuca Sativa” Garden

Lettuce. It is a cultivated variety.

2. Barri (Jungali) “Lactuca Scariola” Wild

Lettuce.

Cultivated variety could be differentiated

into two types. (1) This is 1½ meter high, soft,

smooth and sweetish stem with pinnate wide

leaves, delicately branched which has white

flowers. Seeds are small and whitish in color.

(2) The second harvested variety is English

(Firangi) which is again of two types. Amongst

which, one is well breakable and sweetish.

Harvested Kahu is used as vegetable (Salad).

Wild variety (Barri) of Lactuca Scariola has

longer and thinner leaves than cultivated

variety, the leaves are dark green and slightly

bitter in taste. The latex is sometimes used as



the substitute of opium which is not as potent

as of Khash Khash (Papaver Somniferum)

(Ghani, M.N.K. 1921, Ibne-Sina, 2007).

Description:

It is found as an erect glance sent Annual

or biennial, leafy, 60–150 cm high somewhat

prickly plant at the Western Himalaya from

Marri to Kunawar, at altitude of 6000–11000

feet. It is also found in Western Tibbet at

altitudes of 9000–12000 feet. Distributed to

Siberia and West ward to the British Islets and

canaries .Stem 3–10 cm high, sparingly prickly

below, Capitula is 8–15 m long. Achenes have

6–8 mm body elliptical Setose at apex, 5–9

ribbed, grayish beak as long as body. Leaves

are Pinnatifide or Lobed; achenes light colored,

rarely sinuate, tending to turn edgewise into a

vertical position. Flowers are Yellow, Achenes

striate dark brown or grayish brown. Seeds are

small whitish grey, about 1.0 cm long and 1–2

mm broad (Dymock, W. et al.,1890, Chopra,

R.N. et al.,1956, Kirtikar, K.R et al.,1987).

Unani description

Seeds are whitish or whitish grey, shiny,

elongated and smaller in size. They are

tasteless or have light bitter taste. According to

some Pharmacognostic experts, the seeds are

light and soft. Seeds, Oil, milky fluid that flows

from the stems (Lactucarium) and Dried

Leaves are used as a drug in Unani system of

medicine. The Mizaj (Temperament) of

Tukhme Kahu is described as Cold 2º & Dry 3º.

But there exists a difference of opinion

regarding the gradation in mizaj. A majority of

the authors described the mizaj as cold in 2nd

degree and dry in 3rd

degree (Ghani,

M.N.K.1921, Kareem, N.A.1765, Gulam,

H.1879, Ansari, A.B.H 1885, Haleem, M.A.

1948) while others consider it as cold and dry

in 2nd

degree (Said Mohd. 1973, Hakeem, M.A.

1953, Ibne-Sina 2007).

Phytochemical Studies

The plant contains alkaloids 0.02 %, sugar

and glycosides 6.5 %, volatile oil in traces; fat

2.2 %, Gums 2.16 %; Organic acids 1.06 %,

Carotene 16 mg %, Vitamin B1 22%, Vitamin

C 44 mg %, Vitamin E 32 and Vitamin K 0.2

mg %.The phytochemical investigations of

seeds revealed the presence of alkaloids, the

bitter substance lettuce, oxalic acid,

lactucopicrin (S. R. Baquar, 1989) and

sesquiterpene esters (J. Alberto et al.,

1992).The alkaloid, lactucin, isolated from the

seeds by Dolejs et al., (1958) and Michal et al.,

(1958), exhibited antipyretic activity (V. S.

Agarwal, 1997) and a triterpenoid saponin

isolated from stem possesses antibacterial

activity (R. N. Yadava et al., 2008).

Pharmacological studies

The methanolic extract of Lactuca

scariola was found to possess spasmogenic,

spasmolytic, bronchodilator, and vasorelaxant

activities. The spasmogenic activity may be

attributed to some cholinergic constituents,

whereas spasmolytic effect may be due to

Ca++

channel blocking components that may

cause relaxation of gastrointestinal, tracheal,

and aortic smooth muscles (Khalid hussain

janbaz et al., 2013). L. scariola exhibited dose

dependent potent analgesic activity. Methanolic

extract of L. scariola can produce significant

analgesic activity but failed to show anti-

inflammatory effect (Fayyaz ahmad et al.,

1992). The methanol extracts prepared from

leaves and stems of L. scariola showed

cytotoxic activity against A549, HePG, MCF7

and HCT116 (Eman Elsharkawy1 et al., 2013).

A new triterpenoid saponin has been isolated

from the seeds of Lactuca scariola. This

compound shows antimicrobial activity against

various bacteria and fungi (R. N. Yadava et al.,

2008). The antioxidant activity of Lactuca

scariola (Asteraceae) was investigated by

measuring the radical scavenging effect on

DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical

and found that the methanolic extract of the

aerial parts of Lactuca scariola showed strong

radical scavenging activity (D. K. Kim, 2001).

Lactucin and its derivatives lactucopicrin and

11beta, 13-dihydrolactucin, which are

characteristic bitter sesquiterpene lactones of

Lactuca virosa were evaluated for analgesic

and sedative properties in

mice. Lactucopicrin appeared to be the most

potent analgesic of the three tested compounds.



Lactucin and lactucopicrin, but not 11beta, 13-

dihydrolactucin, also showed sedative

properties in the spontaneous locomotor

activity test (Wesolowska A. et al., 2006). A

Randomized placebo controlled double-blind

trial of Tukhme Kahu (seeds of Lactuca

scariola Linn.) on mixed anxiety depressive

disorder has shown a significant effect in

reducing anxiety and depressive symptoms

(Ghazala javed, et al., 2009). The oil of

Tukhm-e-Kahu (Lactuca scariola seeds) is

used in Industries i.e. soap-making, paints and

varnish (Anonymous, 1962; Ambasta, S.P.

1986). These studies may provide a scientific

basis to validate the traditional use of Tukhme

Kahu in the management of some

gastrointestinal, respiratory, neurological and

vasospastic ailments. The following table

(Table -1) shows the Pharmacological studies

mentioning the Reported action with

references.

AFAL-O-KHAWAS (Pharmacological

actions)

The eminent Unani physician has described

the Afal-o-khawas (Pharmacological Actions)

in detail. It is commonly used as Musakkin

(analgesic), Munawwim (sedative), Mudir baul

(diuretic) and refrigerant. The following table

(Table -2) shows the Afal-o-khawas

(Pharmacological Actions) with Unani and

ethno-medical references.

Therapeutic uses

According to ancient Unani text, Tukhme

kahu and its compound formulations i.e.

roghan Kahu & roghan laboob saba,Mufarreh

shaikur rais etc has been recommended for the

treatment of headache, insomnia, nervousness,

fever, palpitation, burning micturition etc. The

following table (Table -3) shows the

therapeutic uses in different ailments with

Unani and ethno-medical literature references.

Table- 1: Pharmacological studies with references

S.no. Components used Reported action

1. L. sativa L. scariola (whole

plant extract)

CNS stimulant and depressant action respectively in

mice LD50 = 750 mg/kg, IP (Dhawan, B.N., et al.,

1977.)

2. Lactuca scariola

(seeds extract)

Anti-arrhythmic and cardiac function hemodynamic

effect LD50 = 79.05 g/kg, IP (Weizhi, W, et al., 1992)

3. Lactuca virosa

(isolated biologically)

Spontaneous locomotor activity and analgesic effect

active crude preparation P-1, P-2, P-3 and Lactucin)

(Gromek, D et al., 1992)

4. Lactuca sativa

(Seeds oil)

Sedative effects in loco-motor activity, potentiation of

the hypnotic effect of barbiturates, analgesic effect of

barbiturates,( Said Mohd. 1973)

5. Triterpenoid saponin,

isolated from the stems of

L.scariola

Antibacterial triterpenoid saponin (R. N. Yadava et

al., 2008)

6. Methanolic extract of

theaerial parts of Lactuca

scariola

antioxidant activity (D. K. Kim, 2001)

7. Lactucin & lactucopicrin Analgesic and sedative activities in mice

(Wesolowska A. et al., 2006)

8. Methanol extract of L.

scariola

Spasmolytic, bronchodilator, and vasorelaxant

Activities in rabbits (Khalid et al., 2013)

9. Methanolic extract of L. potent analgesic activity in mice (Fayyaz ahmad et



scariola al.,, 1992)

10. Methanolic extracts of leaves

and stems of L. scariola

Anticancer activity (Eman Elsharkawy1 et al., 2013)

11. The extract of Tukhm-e-

kahu

Diuretic, sedative (Ansari A.N, 2009)

12. Tukhm-e-kahu (Lactuca

scariola)

Anxiolytic & antidepressant (Ghazala javed, et al.,

2009)

Table 2: Afal-o-khawas (Pharmacological Actions) with references

S.no

.

Afal-o-khawas

(pharmacologi

cal actions)

Reference

Unani Literature

Reference

Ethno-Medical Literature

1. Hypnotic (Ghani, 1921, Kareem,1765, Gulam,

1879, Ansari, 1885, Haleem, 1948,

Ibne-Baitar (1197-1248 AD), Antaki,

(1597), Attar (1888), Azam, (1895),

Said (1969), Abid, (1907)

Nadkarni 1954, Anonymous 1962,

Dymock (1890), Bunney 1992, Watt,

G. 1972, Khorey, 1985, Chopra

1956, Said, H.M. 1969, Ambasta

1986.

2. Sedative Kareem, 1765; Azam, 1895, Kabir,

1951, Ibne-Sina 2007

Nadkarni, 1954; Waren, 1956,

Chopra, 1956; Watt, 1972 Mair,1973;

Khory, 1985; Bunney 1992;

3. Antipyretic Kabiruddin, H. (1951), Ibne-Sina

(2007)

Anonymous (1962), Khorey, 1985,

Chopra 1956, Kirtikar, 1987, Said,

1969, mbasta, 1986,

4. Diuretic Ghani, 1921; Hakeem, 1953;

Fazlullah, 1970. Ibne-Sina 2007.

Nadkarni, 1954, Chopra, 1956;

Anonymous, 1962, Watt, 1972;

Murray, 1983; Khory, 1985

5. Expectorant Kareem, 1765; Ahmad, 1887;

Ghani,1921; Hakeem, 1953;

Nadkarni, 1954; Chopra 1956;

Khory, 1985;

6. Blood purifier Kabir, 1951; Fazlullah 1970 Dymock, et al..1890

7. Aphrodisiac by

decreasing

anxiety level

Antaki, 1597; Kareem, 1765; Ansari,

1885; Attar, 1888; ; Abid, 1907;

Ghani, 1921; Ibne-e-Sina, 2007,

-

8. Anesthetic Kareem, 1765; Gulam, 1879;

Ansari, 1885; Ghani, 1921;

Haleem, 1948; Hakeem, 1953.

-

9. Antidote Antaki, 1597; Kareem, 1765; -



Attar,1888; Ghani, 1921;

10. Anodyne - Nadkarni,1954; Charles. 1974;

Murry, 1983; Khory, 1985;

11. Anti

inflammatory

Antaki, 1597; Ibn-e-Sina, 2007; -

12. Anti bacterial - R. N. Yadava et al.. 2008.

13. Anti spasmodic - Watt,1972; Chopra,1956;

14. Cooling Kareem, N.A. (1765); Azam, M.K.

(1895) Ibne-Sina (2007)

Dymock,1890; Nadkarni, 1954;

Chopra, 1956; Khory, 1985;

15. Demulcent - Nadkarni, 1954; Chopra, 1956;

Khory, 1985; Kiritikar 1987.

16. Deobstruent Ghani, 1921; Fazlullah, 1970; -

17. Diaphoretic - Chopra, 1956; Watt, 1972; Charles,

1974; Murray, 1983

18. Hair tonic Azam,1895; Kabir,1951; Hakeem

1933

-

19. Purgative - Watt, 1972; Murray, 1983

20. Soothing - Watt, 1972

21. Emmenogogue Kareem, N.A. 1765; Ibne-Sina 2007; -

Table 3: therapeutic uses with references

S.no. Therapeutic uses Reference (Unani & Ethno-Medical Literature)

1. Insomnia Antaki, 1597; Kareem, 1765; Attar, 1888; Ibne Baitar, 1197-1248 Ad;

Azam,1895; Ghani, 1921; Nadkarni, 1954;Anonymous, 1962;

2. Headache Ibne Baitar, 1197–1248 AD; Antaki, 1597; Kareem, 1765;Gulam, 1879;

Ansari, 1885; Ghani, 1921; Haleem, 1948; Hakeem, 1953; Kirtikar &

Basu, 1987.

3. Fever Kabir, 1951; Nadkarni, 1954; Said, 1969; Khory, 1985)



4. Nervousness Nadkarni, 1954; Khory, 1985; Ghazala,2009;

5. Palpitation Nadkarni, 1954; Khory, 1985.

6. Asthma Nadkarni, 1954; Chopra, 1956; Watt, 1972; Murray, 1983;

7. Chronic

bronchitis

(Nadkarni, 1954; Chopra, 1956; Watt, 1972; Kirtikar &

Basu,1987; Ibn Baitar, 1197-1248 AD)

8. Chest pain Kareem, 1765;

9. Acute

inflammation

(Antaki, 1597; Ibne-e-Sina, 1930; Nadkarni, 1954; Khory, 1985; Kirtikar

& Basu, 1987)

10. Burning

micturition

(Kareem, 1965; Gulam, 1889; Ansari, 1885; Ghani,1921.

11. Acute cold/

Coryza

(Kareem, 1765; Ghani, 1921; Hakeem, 1953;

12. Dropsy Murry, 1983;

13. Nocturnal

emission

Antaki, 1597; Ansari, 1885; Attar, 1888; Ghani, 1921; Ibne Sina, 2007;

Nadkarni, 1954; Murry, 1983.

14. Painful irritable

ulcer

Nadkarni, 1954; Ambasta, 1986; Kirtikar Basu, 1987; Ibne Sina, 2007.

15. Pertusis Nadkarni, 1954; Chopra, 1956; Watt, 1972; Murray, 1983.

16. Jaundice Ibne Sina, 2007.

17. Prevent hair fall Kabir, 1951; Anonymous, 1962; Kirtikar & Basu, 1987.

18. Relief of thirst Azam,1895; Ghani,1921; Hakeem,1953; Ibne Sina, 2007.

19. Scorpion sting Antaki,1597; Kareem,1765;Attar,1888;Ghani, 1921; Ibne Sina, 2007

20. Spermatorrhoea Antaki, 1597; Ansari, 1885; Attar, 1888; Ghani, 1921; Ibne Sina, 2007;

Nadkarni, 1954; Murray, 1983.

21. Sunstroke Antaki, 1597, Ibne Sina, 2007

22. Dysmenorrhoea Kareem, 1765; Ibne-e-Sina, 2007; Kirtikar & Basu, 1987; Murry, 1983.



Substitute:

The Majority of authors considered Khash-

khash (Poppy seeds, Papaver somniferum) as

substitute of Tukhm-e-Kahu (Lactuca scariola

seeds). (Kareem, 1765; Ghani, 1921; Hakeem,

1953; Fazulullah, 1970; Ibne Sina, 2007).

While others considered Dammul Akhwain

(Dracaena cinnabin) as its substitute (Ansari,

1885; Kareem, 1765; Ibne Sina, 2007)

Dose:

Tukhme Kahu is prescribed in adult dose of

6–12 gms daily orally (Ghani, 1921; Hakeem

1953). Another reported dose is 3–5 gm

(Ansari, 1888; Kabir, 1951).

Preparation:

Tukhme Kahu is used in form of Safoof

(powder), Joshanda (decoction), single or in

combination with other drugs. It‟s oil used as

Zamad (Liniment) topically.

Harmful Effects:

The herbal drugs may pose harmful affects

either in the form of adverse reactions or drug-

drug interactions due to numerous phyto-

constituents present in each part of a plant.

However no serious health risk is noted till date

but some authors have reported that continuous

use of Tukhme Kahu in high doses can cause

atony of body muscles, Dementia, Amnesia,

Loss of vision and sexual debility (Antaki,

1597; Kareem, 1765; Abid, 1907; Ghani, 1921;

Aziz, 1942; Hakeem, 1953; Ibne Sina, 2007).

Correctives:

Mastagi (Pistacia lentiscus) and Honey are

reported as correctives of Tukhme Kahu

(Kareem, 1765; Ansari, 1885; Haleem, 1948;

Hakeem, 1953, Ibne Sina, 2007).

CONCLUSION

Tukhme Kahu is an important drug in

Unani system of medicine used for ages in the

treatment of headache, insomnia, nervousness,

hypertension, palpitation, fever, Asthma,

Chronic bronchitis, acute cold/Coryza,

Scorpion sting etc. Lactuca scariola has its

own importance as it has Anti cancer

antibacterial, antifungal, Spasmolytic,

bronchodilator and vasorelaxant activities. The

scientific analysis of Tukhme Kahu proves

many of the activities mentioned in Unani

classical literature. Further investigations are

needed to find out the mechanism of action,

active principle(s) and utility of Tukhme Kahu

in clinical practice. Though the drug has been

found to be safe but the potent curative effects

of the drug need to be verified by more

controlled and exhaustive clinical trials,

especially in the field of cancer and

hypertension so that it can be established as a

standard drug.

ACKNOWLEDGEMENT

The author is grateful to Dr Zaki anwar

ansari, Principal, HSZH Govt. Unani medical

college, Bhopal (MP) for his cooperation in the

work. The technical help provided by Mr.

Javed ahmed, library attendant, A & U Tibbia

college (University of Delhi) Karol Bagh New

Delhi-5 and Mrs. Salma, librarian HSZH Govt.

Unani medical college, Bhopal (MP) are also

acknowledged.

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