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An International, Peer Reviewed, Open access, Monthly E-Journal
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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
Ayurveda – Review Article
CRITICAL APPRAISAL SKILL FOR THE ARTICLE PUBLISHED IN AYURVEDIC HEALTH
DOMAIN
Vaikos C D, Waghchoure Ashok 460–464
Unani – Review Article
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
COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – INFLORESCENCE OF ARAGVADHA (CASSIA FISTULA. L. ),
OF THE FAMILY CAESALPINACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT, KARNATAKA, INDIA
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: nataraj_2001us@yahoo.com; 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
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 ||
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.
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 ||
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).
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 ||
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.
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 ||
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
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 ||
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
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 ||
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.
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 ||
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Source of Support: Nil Conflict of Interest: None Declared
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
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: salahmartin@yahoo.fr
Received: 07/04/2013; Revised: 31/05/2013; Accepted: 07/06/2013
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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 410–417
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 410–417
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 410–417
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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
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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
No growth from 1st –
7th
day
Aspergillus
fumingatus
No growth from 1st –
7th
day
No growth from 1st –
7th
day
No growth from 1st –
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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 410–417
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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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Source of Support: Nil Conflict of Interest: None Declared
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
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: rajashekhar@utu.ac.in; rajshekhar.blr@gmail.com; Mob: +919448371037
Received: 20/04/2013; Revised: 23/05/2013; Accepted: 27/05/2013
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,
Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 418–427
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 418–427
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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,
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Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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
EGb 761 standardized
extract
Treatment for
dementia
196 Not
effective
Kanowski et al.,
1996
EGb 761 standardized
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
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Table-2: Selected Clinical Trials Employing Ginkgo on Alzheimer's disease
Reference Plant part /
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
Reference Plant part /
herbal product
Purpose of study Number of
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
Reference Plant part /
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)
Treatment for cerebral
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)
Treatment for cerebral
insufficiency
58 Effective
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Table-5: Selected Clinical Trials Employing Ginkgo on Tinnitus
Reference Plant part /
herbal product
Purpose of study Number of
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
Reference Plant part /
herbal product
Purpose of study Number of
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
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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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Wang, C.W. (1961) The Forests of China,
Maria Moors Cabot Found., publ. 5.
Harvard Univ., Cambridge, Mass.
Woerdenbag H J and De Smet P A G M. (2000)
Adverse effects and toxicity of Ginkgo
biloba extract Med Aromat plants Ind
Profiles; 12: 443–451.
Source of Support: Nil Conflict of Interest: None Declared
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Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
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: drchethankumar@gmail.com; Mob: +919986550745
Received: 07/04/2013; Revised: 21/05/2013; Accepted: 24/05/2013
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,
Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 428–434
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 428–434
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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.
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Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 428–434
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 428–434
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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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 428–434
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
(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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 428–434
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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
Source of Support: Nil Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 435–440
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
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: monika.harmony@gmail.com ; Mob: +91 7838241559
Received: 06/04/2013; Revised: 30/05/2013; Accepted: 04/06/2013
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,
Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 435–440
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 435–440
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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.
MATERIAL AND METHODS
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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 435–440
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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.
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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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 435–440
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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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G and Vanin A (2003) . The influence
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on nitric oxide level and lipid
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therapy of epilepsy, chapter 21 In:
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Source of Support: Nil Conflict of Interest: None Declared
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Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
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: krunaldoshi760@gmail.com; Mob : 09898907572
Received: 04/04/2013; Revised: 23/05/2013; Accepted: 28/05/2013
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,
Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 441–447
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 441–447
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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.
MATERIAL AND METHODS
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
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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
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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
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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
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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.
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Source of Support: Nil Conflict of Interest: None Declared
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
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: drdeeyashri2011@rediffmail.com; Mob +919414893921, +919414458895
Received: 10/05/2013; Revised: 26/05/2013; Accepted: 30/05/2013
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,
Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 448–459
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 448–459
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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).
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Flow chart 1: Probable Etiopathogenesis of Autism
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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
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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-
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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
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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).
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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).
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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
Chaturvedi G. & Shastry
K., 1996
5. Increased connectivity of NE
pathway
Impairment of
pranavayu
Chaturvedi G. & Shastry
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
Chaturvedi G. & Shastry
K., 1996
8. Excessive storage of all information
Based on principle of
“ samanyam vriddhi
karanam”
Chaturvedi G. & Shastry
K., 1996
9. Hyper memory Increased Udanvayu Arunadutta, 2002
10. Hyper preferential pathway Excessive intake of
smritihetu
Chaturvedi G. & Shastry
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
Chaturvedi G. & Shastry
K., 1996
14. Lack of communication and social
Impairment
Impairment of Udan vayu
and Sadhak pitta
Arunadutta, 2002,
Chaturvedi G. & Shastry
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
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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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(2001). Unusual brain growth patterns
in early life in patients with
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Goldsmith, H.H., Alexander, A.L. and
Davidson, R.J. (2005). Gaze fixation
and the neural circuitry of face
processing in autism. Nat. Neurosci., 8,
519–526
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intact global processing among
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B. and Burack, J. (2006). Enhanced
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Rinaldi, T., Perrodin, C. and Markram, H.
(2008). Hyper-connectivity and hyper-
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Source of Support: Nil Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 460–464
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
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 - vaikosc@gmail.com
Received: 10/05/2013; Revised: 26/05/2013; Accepted: 02/06/2013
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,
Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 460–464
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 460–464
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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
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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
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 460–464
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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.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 460–464
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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
Source of Support: Nil Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 465–474
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
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: drarifmd@gmail.com
Received: 20/04/2013; Revised: 23/05/2013; Accepted: 27/05/2013
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,
Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 465–474
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 465–474
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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
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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.
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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
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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; -
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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)
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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.
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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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