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Four groups of Swiss albino mice (male) were administered with vehicle (refined vegetable oil), 1/20 th , 1/10 th and 1/5 th of LD 50 doses of methanolic extract of Eupatorium adenophorum Spreng ; respectively for a period of 21 days. Mice fed with methanolic extract of Eupatorium adenophorum at a dose level of 752.2 mg/kg (i.e. 1/5 th LD 50) elicited hepatotoxicity and the animals had yellow discoloration of liver, subcutaneous tissue and musculature indicating jaundice. The sera samples revealed marked increase in bilirubin levels and activities of alkaline phos-phatase (ALP), alanine transaminase (ALT), aspartate transaminase (AST). Histopathology of the livers of the group IV animals had focal areas of necrosis and bile duct proliferation. Elevation in plasma bilirubin concomitant with alterations in enzyme profile and histopathological lesions are consistent with liver injury and cholestasis.
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Damodar Singh et al., Int. J. Res. Phytochem. Pharmacol., 1(3), 2011, 165-171
©JK Welfare & Pharmascope Foundation | International Journal of Research in Phytochemistry & Pharmacology 165
Short-term Toxicity Studies of Eupatorium adenophorum in Swiss Albino Mice
Y. Damodar Singh*1, S.K. Mukhopadhayay2, M. Ayub Ali3, T.C. Tolenkhomba4 and M.A. Ayub Shah5
1Department of Veterinary Pathology, 3Department of Physiology & Biochemistry; 4Department of Animal Breed-
ing & Genetics and 5Department of Pharmacology & Toxicology College of Veterinary Sciences & A.H., Central
Agricultural University, Selesih, Aizawl, Mizoram, India
2Department of Veterinary Pathology, F/O Veterinary & Animal Sciences, WBUAFS, Kolkata-37, India
ABSTRACT
Four groups of Swiss albino mice (male) were administered with vehicle (refined vegetable oil), 1/20 th, 1/10th and
1/5th of LD50 doses of methanolic extract of Eupatorium adenophorum Spreng ; respectively for a period of 21
days. Mice fed with methanolic extract of Eupatorium adenophorum at a dose level of 752.2 mg/kg (i.e. 1/5th LD50)
elicited hepatotoxicity and the animals had yellow discoloration of liver, subcutaneous tissue and musculature
indicating jaundice. The sera samples revealed marked increase in bilirubin levels and activities of alkaline pho s-
phatase (ALP), alanine transaminase (ALT), aspartate transaminase (AST). Histopathology of the livers of the group
IV animals had focal areas of necrosis and bile duct proliferation. Elevation in plasma bilirubin concomitant with
alterations in enzyme profile and histopathological lesions are consistent with liver injury and cholestasis.
Keywords: Eupatorium adenophorum; Hepatotoxicity; bilirubin; mice
INTRODUCTION
Eupatorium adenophorum Spreng (syn. Ageratina ade-
nophora, common name: Crofton weed; Sticky snake-
root), a native of Central America has appeared as a
major weed in several areas in different parts of the
world and has infested the grazing areas in the lower
and mid hills in the Himalayan region of India (Sharma
et al., 1998). E. adenophorum is widely growing shrub
in most parts of NE region, in general and Mizoram, in
particular. E. adenophorum is an important weedy co-
lonizer in early succession communities developing
after slash and jhum (shifting cultivation) at high eleva-
tions of North Eastern Hill Region of India (Kamakrish-
nan and Mishra, 2006 and Shah, 2007). It is a perennial
herb, nearly 1 meter high and erects (Mandal et al.,
2005). Grazing animals get accidentally exposed to the
plant under scarcity conditions. A considerable varia-
tion between the animal species exists in terms of sus-
ceptibility to toxicity due to E.adenophorum.
E. adenophorum has been investigated for its analges-
ic, anti-inflammatory and antipyretic properties (Bijar-
gi, 2009). The methanolic extract of E. adenophorum
exerted analgesic (Bijargi et al., 2009; Bijargi and Shah,
2010), anti-inflammatory (Bijargi and Shah, 2010, Bijar-
gi et al., 2010) and antipyretic (Bijargi and Shah, 2010)
effects in rats.
There is a lot of variation in the susceptibility of differ-
ent animal species to the noxiousness of E. adenopho-
rum (O’Sullivan, 1979 and Sharma, 1998). The content
of natural products in the plants is known to vary with
geographical region, soil and other environmental fac-
tors (Smith et al., 1994; Kaul and Vats, 1998). Con-
sumption of E.adenophorum by horses results in pul-
monary toxicity (Jones, 1954; O’Sullivan, 1979). Regular
ingestion of Eupatorium adenophorum [Ageratina ade-
nophora (Spreng.)] or Crofton weed causes chronic
pulmonary disease in horses mainly in Australia, New
Zealand, and the Himalayas (Oelrichs et al., 1995).
Toxicity due to consumption of this plant by other graz-
ing animals is not clear. However, no toxic effects were
seen in goats when E.adenophorum collected from
Nepal comprising up to 67% of their intake, was admi-
nistered (Neopane et al., 1992). Experimental feeding
of E.adenophorum plant growing in north-eastern India
to cattle caused anorexia, suspension of rumination
and photosensitization (Verma et al., 1987). In studies
with laboratory animals, mice were shown to be suita-
ble test animals, but in this species lesions occur in the
liver rather than the lungs. Exposure of mice to feed
containing E.adenophorum freeze-dried leaf powder
caused hepatotoxicity. In mice, hepatotoxicity involved
multiple areas of focal necrosis of the parenchyma a s-
sociated with degeneration and loss of epithelial cells
lining of the bile duct (Sani et al., 1992). E. adenopho-
rum leaf samples collected from Kangra Valley (India)
and partially purified extracts from leaf samples mixed
in the diet caused hepatotoxicity and cholestasis in rats
(Katoch et al., 2000 and Kaushal et al., 2001). We re-
port here the LD50 of E. adenophorum (methanolic ex-
tract) and its toxic effects in laboratory mice.
www.ijrpp.pharmascope.org
ISSN: 2231-010X
Research Article
* Corresponding Author
Email: drmaasvptcau@yahoo.co.in
Contact: +91-9436151974
Received on: 13-07-2011
Revised on: 27-07-2011
Accepted on: 28-07-2011
Damodar Singh et al., Int. J. Res. Phytochem. Pharmacol., 1(3), 2011, 165-171
166 ©JK Welfare & Pharmascope Foundation | International Journal of Research in Phytochemistry & Pharmacology
MATERIALS AND METHODS
Collection, identification and extraction of the plant
material
The entire plant of Eupatorium adenophorum Spreng
was collected in April 2010 from the campus of the
College of Veterinary & A.H., Central Agricultural Uni-
versity, Selesih, Aizawl, Mizoram (India) and submitted
as a herbarium specimen for authentication to the Re-
gional Office, Botanical Survey of India (BSI), Shillong.
The BSI, Shillong has authenticated the plant as such by
Letter Reference No.BSI/ERC/Tech/2010/052, dated
27.04.2010.
Figure 1: Leaves and flowers of Eupatorium
adenophorum
The fresh leaves of the plant Eupatorium adenophorum
was carefully collected from the College campus,
washed gently till the leaves looked clean, mopped by
blotting paper and weighed. Then the plant leaves
were air dried under shade, protecting them from di-
rect sunlight for a period of one week. On complete
drying, whole of the leaves were ground to powder
with the instrument Willey grinder and sifted through
sieve number 22. The dry powder of Eupatorium ade-
nophorum was subjected to cold maceration technique
using methanol as solvent as follows:
The cold methanolic extract of E. adenophorum was
prepared as per standard method (Manjunatha et al.,
2005; Harborne, 1998). One hundred (100) grams of
whole plant powder was soaked in 500 ml of methanol
(1: 5 w/v) in a conical flask for 3 days with intermittent
stirring and at the end of 3rd day the content was fil-
tered with muslin cloth followed by Whatman filter
paper No-1. For complete extraction of the active prin-
ciples, this process was repeated three times using
fresh solvent on each occasion or until the colour of
the methanol became light. The filtrate obtained was
pooled and further subjected to vacuum evaporation
at 300C in a Rotary Evaporator and lyophilized for suc-
cessive 24 hours. Lyophilization was stopped when the
extract appeared sufficiently dry. Further the material
was stored at -45oC in deep freezer in air tight contain-
ers until use. The yield of the extract was 29.53 g per-
cent.
Preparation of oral suspension
The methanolic extract was found insoluble in water;
therefore, for different dose levels, a stock suspension
was prepared in refined vegetable oil and was diluted
with the vehicle (refined vegetable oil) immediately
before use for oral administration.
Experimental animals
In the present study, 50 male Swiss albino mice (Mus
musculus) of 25-30 g were obtained from the Colony
Stock of Laboratory Animal House, College of Veteri-
nary Sciences & A.H., Central Agricultural University,
Selesih, Aizawl, Mizoram. They were given a standard
pelleted diet and water ad libitum throughout the ex-
perimental period. A twelve-hour day and night cycle
was maintained in the animal house. The ambient
temperature and relative humidity during the experi-
mental period were 22-24oC and 65-70%, respectively.
The experimental protocol met regulatory guidelines
on the proper care and use of animals in laboratory
research.
Acute toxicity study
Thirty (30) male mice were randomly selected and di-
vided into six groups of five animals each. The animals
were fasted overnight. Group-I animals were orally
administered the vehicle (refined vegetable oil), while
the animals of Groups II-VI were given single doses of
methanol extract of E. adenophorum in progressively
increased manner (1350, 2025, 3050, 4575 and 6900
mg/Kg respectively) for determination of the acute
lethal dose (LD50). However, food and water were pro-
vided throughout the experiment. Immediately after
dosing, the animals were observed continuously for
the first 72 hours for mortality and any signs of overt
toxicity. The surviving animals were also observed up
to 14 days for signs of toxicity. The number of mice
that died within the period of study was noted for each
group, and subsequently the LD50 value calculated (Mil-
ler and Tainter, 1944). All animals that died during the
observation period and euthanatized mice were sub-
jected to necropsy.
Sub-acute toxicity study
Twenty (20) male mice were randomly divided into
four groups of five animals each. Animals of Group-I
served as vehicle (refined vegetable oil) treated con-
trols, while animals of Groups II, III and IV were admi-
nistered orally with the E. adenophorum extract at dai-
ly doses of 188 mg/kg (1/20th LD50), 376 mg/kg (1/10th
LD50) and 752 mg/kg (1/5th LD50) respectively for 21
days. Food and water were freely available during the
experiment. The animals in treated groups were ob-
served daily for physical and behavioural changes as
signs of toxicity. On termination of the experiment, all
the animals were weighed and then euthanized using
ether anesthesia. The blood samples were collected by
retro-orbital plexus into test tubes with the aid of a
capillary tube and then centrifuged at 4000 rpm for 10
Damodar Singh et al., Int. J. Res. Phytochem. Pharmacol., 1(3), 2011, 165-171
©JK Welfare & Pharmascope Foundation | International Journal of Research in Phytochemistry & Pharmacology 167
minutes to separate the serum for estimation of blood
biochemical parameters. Gross lesions present in the
liver, lungs, kidneys, heart and spleen were recorded
and tissues showing lesions were fixed in 10% neutral
buffered formalin immediately for further histopatho-
logical examination.
Biochemical profile
Biochemical parameters; viz. blood glucose, serum
cholesterol, triglycerides, total protein, albumin, total
bilirubin, direct (Conjugated) bilirubin, alkaline phos-
phatase (ALP), alanine transaminase (ALT), aspartate
transaminase (AST), blood urea nitrogen (BUN) and
creatinine were estimated spectrophotometrically
(Spectroscan 2600 Chemito) using standard commer-
cial kits (Crest Biosystems, Goa, India).
Histopathological examination
The formalin-fixed tissues (2-3 mm thick) were taken,
washed overnight in running tap water and then dehy-
drated in ascending grades of alcohol starting from
50%, 70%, 90% and absolute alcohol I, alcohol II, alco-
hol III and finally cleared in cederwood oil or xylene.
These dehydrated tissue pieces were then embedded
in molten paraffin. Sections were cut at 3-5 μm thick
and stained with Mayer’s hematoxylin and eosin for
histopathological examinations (Bancroft and Stevens,
1980).
Statistical analysis
All the values were expressed as mean±SEM. Statistical
analysis was done using SYSTAT 6.0.1 version. The sta-
tistical significance of differences between the two
means was assessed by unpaired Student’s ‘t’ test. A
difference at P<0.05 was considered statistically signifi-
cant.
RESULTS AND DISCUSSION
Acute toxicity
Mice administered with E. adenophorum extract at the
dose level of 1350 mg/kg body weight showed no mor-
tality, while those at dose levels of 2025 and 3050
mg/kg body weight showed partial loss of appetite,
dullness and depression with 20% and 40% mortality
respectively in 24 hours. The dose level of 4575 mg/kg
body weight produced tremor, hypo-activity and 60%
mortality. However, the dose level of 6900 mg/kg body
weight had severe clinical signs and all animals died
within 4-6 hours.
The doses of LD50 study thus obtained were then plot-
ted on semi-logarithmic paper against the probit and a
best fitted linear scale was drawn. The oral acute LD50
of E. adenophorum extract was then determined from
the straight line and was found to be 3761 mg/kg body
weight (3364 ≤ 3761 ≥ 4157 mg/Kg with 95% confi-
dence). This LD50 value was lower than 5000 mg/kg
reported by other workers in mice with alcoholic ex-
tract of E. adenophorum Sprengel (Gao Ping et
al.,2005). However, methanolic extract of E. adeno-
phorum at 2000 mg/kg, die not produce any signs of
overt toxicity in rats (Bijargi et al., 2009). The differ-
ence in the LD50 values might be due to using of differ-
ent vehicle and also due to variation of geographical
region, soil and other environmental factors. This sug-
gests that the E. adenophorum plant growing in the
region is apparently more toxic.
Sub-acute toxicity
Clinical signs
The Group-I (vehicle control) mice remained normal
throughout the experimental period, while the animals
in Group-II (1/20th LD50 i.e. 188 mg/kg) showed a partial
Table 1: Changes in biochemical parameters of mice intoxicated with E. adenophorum methanolic
extract (21 days exposure).
Parameters
Group-I
(Vehicle
control)
Group-II
(188 mg/kg)
Group-III
(376 mg/kg)
Group-IV
(752 mg/kg)
Glucose (mg/dl)
100.43±3.44b
95.65±4.73b
113.29±3.57a
112.99±3.30a
Cholesterol (mg/dl)
181.01±6.51
192.64±8.15
186.63±4.72
185.21±3.43
Triglycerides (mg/dl)
107.68±3.17
107.21±4.00
101.73±3.62
116.68±6.70
Total protein (g/dl)
6.28±0.43
6.53±0.53
6.82±0.39
6.62±0.35
Albumin (g/dl)
4.33±0.36
4.71±0.31
4.44±0.21
4.12±0.20
Total bilirubin (mg/dl)
0.958±0.031d
1.27±0.058c
1.57±0.082b
3.47±0.078a
Conjugated bilirubin (mg/dl)
0.778±0.016d
1.17±0.038c
1.44±0.06b
3.25±0.06a
AST (IU/L)
33.80±1.49c
40.32±1.75c
68.40±2.08b
146.63±6.60a
ALT (IU/L)
43.65±2.45d
56.40±1.86c
91.29±4.25b
265.13±6.58a
Alkaline phosphatase (IU/L)
28.50±1.74c
37.05±1.86b
42.20±1.36b
112.14±3.06a
LDH (IU/L)
114.36±4.61c
103.13±4.75c
175.30±6.57b
453.94±9.56a
Urea (mg/dl)
43.05±1.43
44.17±1.91
42.32±1.06
40.89±1.32
Creatinine (mg/dl)
0.486±0.035
0.534±0.013
0.496±0.025
0.530±0.029
Values are Mean±SE. ** Significance at p ≤ 0.01; * p ≤ 0.05 and NS = Not significant
Damodar Singh et al., Int. J. Res. Phytochem. Pharmacol., 1(3), 2011, 165-171
168 ©JK Welfare & Pharmascope Foundation | International Journal of Research in Phytochemistry & Pharmacology
loss of appetite, dullness and slight depression. The
Group-III animals (1/10th LD50 i.e. 376 mg/kg) became
dull, depressed and had rough hair coat after 10 days
of treatment. However, the animals in Group-IV (1/5th
LD50 i.e. 752 mg/kg) became dull, depressed and ano-
rectic within 10 days and leading to body weight loss
(data not shown). The mice from this group had rough
hair coat and appeared jaundiced when observed after
7 days of exposure. The ear pinnae and paws became
yellowish. The plants belonging to the genus Eupato-
rium are reported to be hepatotoxic and pneumotoxic
in horse (O’Sullivan, 1979)
Changes in the biochemical parameters
The effect of methanolic extract of E. adenophorum
(Spreng) on the biochemical parameters is summarized
in Table 1. The increase in serum glucose levels of
Groups III (376 mg/kg) and IV (752 mg/kg) animals
were highly significant (P <0.01) as compared to those
in Group-I (vehicle control) and Group-II (188 mg/kg).
No significant changes were observed in the levels of
cholesterol, triglycerides, total protein and albumin in
all other experimental groups. The total bilirubin levels
as well the conjugated bilirubin were much higher (P
<0.01) in the Group IV as compared to those in the
Groups II, III and the control. These observations pro-
vide the biochemical basis for the observed yellowish
discolouration of liver and the epidermis. The increase
in the bilirubin levels was more marked in the conju-
gated form which is characteristic of obstructive jaun-
dice and cholestasis (Cornelius, 1980). There was
marked increase (P<0.01) in the activities of aspartate
aminotransferase (AST), alanine aminotransferase
(ALT), alkaline phosphatase (ALP) and lactate dehydro-
genase (LDH) in the animals in Group III and IV as
compared to those in Group-I (control) and Group-II.
The increase in the activity of transaminases is known
to be the indicators of degenerative changes in organs
or tissues like liver and myocardium (Kaneko, 1980;
Cornelius, 1989 and Neiger and Osweiler, 1989). In-
creased levels of transaminases and ALP activities are
also known to occur in a wide range of diseases of liver
like cholestasis, biliary obstruction and hepatic necrosis
(Tennant, 1997). The elevation of serum enzymatic
activities in the present study is attributable to E. ade-
nophorum -induced hepatic damage and/or necrosis as
confirmed from histopathological observations.
There were no changes in the levels of blood urea and
creatinine in all the E. adenophorum (methanolic ex-
tract)- treated groups as compared to the control indi-
cating that there was no nephrotoxic effect of metha-
nolic extract of E. adenophorum in mice when exposed
for 21 days.
Similar biochemical changes have been observed in the
plasma of rats exposed to leaf powder, methanolic
extract and partially purified fraction of E. adenopho-
rum (Sharma et al., 1998; Katoch et al., 1999; Kaushal
et al., 2000 & 2001 and Bhardwaj et al., 2001).
Figure 2: A) Livers of mice from groups I, II and III showing no appreciable cross changes and that of group
IV showing marked enlargement with yellowish discoloration. B) Postmartum examination of a mouse
from group IV showing yellowish discoloration of liver, subcutaneous tissue and musculature indicating
severe jaundice.
Damodar Singh et al., Int. J. Res. Phytochem. Pharmacol., 1(3), 2011, 165-171
©JK Welfare & Pharmascope Foundation | International Journal of Research in Phytochemistry & Pharmacology 169
Gross and histopathological changes
Postmortem examination of the animals in the groups
I, II and III revealed no appreciable gross changes of the
liver and other visceral organs, while the animals of the
group IV which received 1/5th LD50 (752.2 mg/kg) of E.
adenophorum extract had yellowish coloration of liver,
subcutaneous tissue and musculature (Fig. 2 and 3) and
the urinary bladder was full of urine.
Histopathological studies also provided supportive evi-
dence for the biochemical analysis depicted by the
following photomicrographs. Figs. 3a and 3b showed
the normal architecture and mild degenerative
changes of liver with narrow sinusoidal spaces in
Groups I (control) and II (188 mg/kg) animals respec-
tively. Liver sections of Group III (376 mg/kg) animals
revealed mild to moderate bile duct proliferation and
focal areas of necrosis (Fig. 3c). No histopathological
lesions were observed in other tissues collected.
In Group-IV (752 mg/kg), the liver sections showed
dilatation of the bile ducts and proliferative changes
with infiltration of mononuclear cells (Fig.3d). The he-
patocytes around the bile ducts showed necrotic
changes as well as some focal areas of necrosis (Fig.
3d). Similar changes have been observed toxicity in-
duced by the whole leaf powder, methanolic extract
and partially purified fraction of E. adenophorum in
rats (Sharma et al., 1998; Katoch et al., 1999; Kaushal
et al., 2000 & 2001 and Bhardwaj et al., 2001). Bile
canalicular plasma membrane has been proposed as
the primary site of biochemical lesions in hepatotoxici-
ty induced by lantana toxins (Pass et al., 1978; Sharma
and Dawra, 1984). Other workers also observed dege-
neration of intrahepatic bile ducts and hepatocellular
necrosis following of administration of E. adenophorum
leaf powder to mice (Sani et al., 1989).
The present study shows that the toxicity of E. adeno-
phorum (methanolic extract) is dose dependant and
the dose level of 1/5th LD50 (752 mg/kg body wt) is
highly hepatotoxic in mice. Therefore, E. adenophorum
should be avoided from feeding to livestock as it could
be potentially toxic to higher animals too. Further long
term toxicity studies and on other toxicological aspects
on E. adenophorum are advocated.
ACKNOWLEDGEMENTS
The authors are thankful to the Dean, College of Vete-
rinary Sciences & A.H., Central Agricultural University,
Selesih, Aizawl, Mizoram for providing the facilities of
the College to conduct the experiment.
Fig. 3d
Fig. 3a
Fig. 3b
Fig. 3c
Fig. (3a) Control liver showing normal architecture (H&E x 200); (3b) Group-II liver showing mild dege-
nerative changes and narrowing of sinusoidal spaces of hepatocytes (H&E x 200); (3c) Group-III liver
showing mild to moderate bile duct proliferation
Damodar Singh et al., Int. J. Res. Phytochem. Pharmacol., 1(3), 2011, 165-171
170 ©JK Welfare & Pharmascope Foundation | International Journal of Research in Phytochemistry & Pharmacology
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... Scientific RepoRts | 5:15967 | DOi: 10.1038/srep15967 methanolic extract of E. adenophorum leaf samples induces hepatotoxicity in albino mice 10 . ...
... Scientific RepoRts | 5:15967 | DOi: 10.1038/srep15967 methanolic extract of E. adenophorum leaf samples induces hepatotoxicity in albino mice 10 . Furthermore, feeding rats a diet mixed with purified extracts from E. adenophorum leaf samples causes hepatotoxicity and cholestasis 11,12 , and previous studies have shown that the active Euptox A isolated from E. adenophorum represents an important toxin showing hepatotoxicity 5,13 . ...
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The precise cytotoxicity of E. Adenophorum in relation to the cell cycle and apoptosis of splenocytes in Saanen goats remains unclear. In the present study, 16 Saanen goats were randomly divided into four groups, which were fed on 0%, 40%, 60% and 80% E. adenophorum diets. The results of TUNEL, DAPI and AO/EB staining, flow cytometry analysis and DNA fragmentation assays showed that E. adenophorum induced typical apoptotic features in splenocytes, suppressed splenocyte viability, and caused cell cycle arrest in a dose-dependent manner. However, westernblot, ELISA, qRT-PCR and caspase activity analyses showed that E. adenophoruminhibited Bcl-2 expression, promoted Bax translocation to the mitochondria, triggered the release of Cytc from the mitochondria into the cytosol, and activated caspase-9 and -3 and the subsequent cleavage of PARP. Moreover, in E. adenophorum-induced apoptosis, the protein levels of Fas, Bid, FasL and caspase-8 showed no significant changes. E. adenophorum treatment induced the collapse of δψ m. Moreover, these data suggested that E. adenophorum induces splenocyte apoptosis via the activation of the mitochondrial apoptosis pathway in splenocytes. These findings provide new insights into the mechanisms underlying the effects of E. adenophorum cytotoxicity on splenocytes.
... E. adenophorum has pneumotoxic and hepatotoxic effects on animals, especially horses being more susceptible to its toxicity. Consumption of leaves of this plant by horses causes a chronic pulmonary disease known as Numinbah Horse Sickness, whereas in goats no effect has been observed after ingestion in the Nepal Himalayas [66,67]. It causes anorexia and photosensitization in cattle. ...
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Indian Himalayan region (IHR) supports a wide diversity of plants and most of them are known for their medicinal value. Humankind has been using medicinal plants since the inception of civilization. Various types of bioactive compounds are found in plants, which are directly and indirectly beneficial for plants as well as humans. These bioactive compounds are highly useful and being used as a strong source of medicines, pharmaceuticals, agrochemicals, food additives, fragrances, and flavoring agents. Apart from this, several plant species contain some toxic compounds that affect the health of many forms of life as well as cause their death. These plants are known as poisonous plants, because of their toxicity to both humans and animals. Therefore, it is necessary to know in what quantity they should be taken so that it does not have a negative impact on health. Recent studies on poisonous plants have raised awareness among people who are at risk of plant toxicity in different parts of the world. The main aim of this review article is to explore the current knowledge about the poisonous plants of the Indian Himalayas along with the importance of these poisonous plants to treat different ailments. The findings of the present review will be helpful to different pharmaceutical industries, the scientific community and researchers around the world.
... E. adenophorum has pneumotoxic and hepatotoxic effects on animals, especially horses being more susceptible to its toxicity. Consumption of leaves of this plant by horses causes a chronic pulmonary disease known as Numinbah Horse Sickness, whereas in goats no effect has been observed after ingestion in the Nepal Himalayas [66,67]. It causes anorexia and photosensitization in cattle. ...
Article
Full-text available
Indian Himalayan region (IHR) supports a wide diversity of plants and most of them are known for their medicinal value. Humankind has been using medicinal plants since the inception of civilization. Various types of bioactive compounds are found in plants, which are directly and indirectly beneficial for plants as well as humans. These bioactive compounds are highly useful and being used as a strong source of medicines, pharmaceuticals, agrochemicals, food additives, fragrances, and flavoring agents. Apart from this, several plant species contain some toxic compounds that affect the health of many forms of life as well as cause their death. These plants are known as poisonous plants, because of their toxicity to both humans and animals. Therefore, it is necessary to know in what quantity they should be taken so that it does not have a negative impact on health. Recent studies on poisonous plants have raised awareness among people who are at risk of plant toxicity in different parts of the world. The main aim of this review article is to explore the current knowledge about the poisonous plants of the Indian Himalayas along with the importance of these poisonous plants to treat different ailments. The findings of the present review will be helpful to different pharmaceutical industries, the scientific community and researchers around the world.
... According to Liu et al. [14], A. adenophora synthesizes numerous bioactive compounds. A number of studies have been conducted to identify, isolate and study these phytochemicals and the responses of plants and animals to these allelochemicals [15][16][17][18][19][20][21][22][23][24] According to Bhardwaj et al. [25] and Thapa et al. [26], maximum quantities of phytotoxins are synthesized and present in the leaves of A. adenophora. These chemicals are released by the plants in the form of either leachates, root secretion or via decomposition of plant materials and provide them the competitive advantage over the co-occurring vegetation leading to the removal of previously growing species. ...
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Ageratina adenophora is among the most appalling invasive alien weeds spreading over the majority of habitats in tropical and sub-tropical countries. Agricultural lands are one of the major invaded areas by this weed and its luxuriant growth has been reported along the croplands. To lessen the harmful impacts posed by this weed, farmers and land managers usually slash the aerial parts of this weed. But the piled residues are not treated and are left which could have the impacts on the crops growing in its vicinity. In this study, impacts of fresh, dry and composted leaf tissues of A. adenophora in pot experiment against two food crops viz. Triticum aestivum (wheat, commonly known as gehu) and Lens culinaris (lentil, commonly known as masoor) were investigated. Five concentration treatments i.e. C0 (control-0 g leaves/kg soil)), C1 (10g leaves/kg soil), C2 (20g leaves/kg soil), C3 (40g leaves/kg soil) and C4 (80g leaves/kg soil) were prepared and the effects on germination, growth (shoot and root), dry matter accumulation and yield of test plant species were recorded. Fresh and dry leaf treatments inhibited the performance of lentil as well as wheat while the composted leaves promoted seed germination, plant height, biomass and crop yield in concentration dependent manner. As compared to fresh and composted leaves, test crops were severely affected by dry leaf treatments and failed to produce pods and seeds towards higher concentrations. The results revealed that fresh and dry leaves of A. adenophora have inhibitory effect on germination and growth of test plants while composting of A. adenophora leaves reduced the inhibitory effect of weed residues incorporated into the soil.
... Regular consumption of crofton weed by horses leads to chronic lung disease, known as Numinbah Horse Sickness or Tallebudgera Horse Disease in northern New South Wales and Queensland. However, no toxic effects were seen in goats with ingestion of plant from Nepal [95]. Experimental feeding of E. adenophorum plant growing in north-eastern India to cattle caused anorexia, suspension of rumination and photosensitization [96]. ...
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Eupatorium adenophorum Spreng belonging to the family Asteraceae have traditionally been used as folklore medicine across the world. In traditional system of medicine, it is regarded as anti- inflammatory, antimicrobial, antiseptic, analgesic, antipyretic, blood and coagulant. The present review summarizes the updated information concerning the ethnomedicinal, phytochemical, pharmacological and toxicological aspects of E. adenophorum. A thorough bibliographic investigation was carried out by analyzing worldwide accepted scientific data base (Pub Med, Google Scholar, Scopus SciFinder, and Web of Science), thesis, recognized books and other accessible literature from 1980 to 2017. The phytochemical and pharmacological studies demonstrated that E. adenophorum possess a wide spectrum of pharmacological activities, such as anti-inflammatory, analgesic, antipyretic, antioxidant, antibacterial, antifungal, antitumor, antioxidant, antiseptic and cytotoxic activities which could be attributed to the presence of array of phytochemicals of various groups including terpenoids, phytosterols, alkaloids, flavonoids, phenolic acids, coumarins, phenylpropanoids, sesquiterpene lactones, polysaccharides, and essential oil. Modern phytochemical and pharmacological studies have led to the isolation and characterization of a number of bioactive compounds from different parts of the plant as well as validation of its traditional medicinal uses. However, certain known toxic effects of E. adenophorum demand a thorough study of long-term toxicity and other toxicological aspects. Furthermore, the relationship of molecular structures of compounds of E. adenophorum with its various pharmacological activities needs further study and confirmation.
... Experimental feeding of E. adenophorum plant growing in north-eastern India to cattle caused anorexia, suspension of rumination and photosensitization [96]. Methanolic extract of E. adenophorum leaf samples collected from Mizoram (India) has also been reported to induce hepatotoxicity in albino mice [97]. Pathological findings include pulmonary interstitial fibrosis and alveolar epithelisation. ...
Preprint
Full-text available
Eupatorium adenophorum Spreng belonging to the family Asteraceae have traditionally been used as folklore medicine in different parts of the world. In traditional system of medicine, it is regarded as anti-inflammatory, antimicrobial, antiseptic, analgesic, antipyretic, blood and coagulant. The present review summarizes the updated information concerning the ethnomedicinal, phytochemical, pharmacological and toxicological aspects of E. adenophorum. A thorough bibliographic investigation was carried out by analyzing worldwide accepted scientific data base (Pub Med, Google Scholar, Scopus SciFinder, and Web of Science), thesis, recognized books and other accessible literature from 1980 to 2017. The phytochemical and pharmacological studies demonstrated that E. adenophorum possess a wide spectrum of pharmacological activities, such as anti-inflammatory, analgesic, antipyretic, antioxidant, antibacterial, antifungal, antitumor, antioxidant, antiseptic and cytotoxic activities which could be explained by the presence of bioactive chemical constituents including terpenoids, phytosterols, alkaloids, flavonoids, phenolic acids, coumarins, phenylpropanoids, sesquiterpene lactones, polysaccharides, and essential oil. Modern phytochemical and pharmacological studies have led to the isolation and characterization of a number of bioactive compounds from different parts of the plant as well as validation of its traditional medicinal uses. However, certain known toxic effects of E. adenophorum demand a thorough study of long-term toxicity and other toxicological aspects. Furthermore, the relationship of molecular structures of compounds of E. adenophorum with its various pharmacological activities needs further study and confirmation.
... Furthermore, the rats administrated with purified extracts from E. adenophorum leaves as a diet supplement exhibited hepatotoxicity and cholestasis [18]. The leaves of E. adenophorum contain abundant sesquiterpenes [18][19][20][21]. Among them, 9-oxo-10,11-dehydroageraphorone is considered as the main toxin of E. adenophorum. ...
Article
Full-text available
Eupatorium adenophorum is widely distributed throughout the world’s tropical and temperate regions. It has become a harmful weed of crops and natural environments. Its leaves contain bioactive compounds such as chlorogenic acid and may be used as feed additives. In this study, chlorogenic acid was extracted and separated from leaves of E. adenophorum. Three chlorogenic acid products were prepared with different purities of 6.11%, 22.17%, and 96.03%. Phytochemical analysis demonstrated that the main toxins of sesquiterpenes were almost completely removed in sample preparation procedure. The three products were evaluated for safety via in vitro and in vivo toxicological studies. All the products exhibited no cytotoxic effects at a dose of 400 μg/mL in an in vitro cell viability assay. When administered in vivo at a single dose up to 1.5 g/kg bw, all three products caused no signs or symptoms of toxicity in mice. These results encourage further exploration of extracts from E. adenophorum in feed additive application.
... Previous study indicated that E. adenophorum has the acaricidal activity [6][7][8], antitumor activity [9,10], potential anti-inflammatory and other biological activities [11]. The previous reports showed that purified extracts from E. adenophorum could cause hepatotoxicity of mice and livestock [8,[12][13][14][15], especially the 9-oxo-10,11-dehydro-agerophorone which is the main poisonous components of E. adenophorum [16,17]. Also, when administrated with E. adenophorum freeze-dried leaf powder, the mice were caused hepatotoxicity. ...
Article
Full-text available
E. adenophorum has reported to cause hepatotoxicity. But, the precise effects of E. adenophorum on hepatocytes is unclear. Saanen goats were fed on E. adenophorum to detect the cytotoxicity effects of E. adenophorum on hepatocytes. Our study has shown that the typical apoptotic features, the increasing apoptotic hepatocytes and activated caspase-9, -3 and the subsequent cleavage of PARP indicated the potent pro-apoptotic effects of E. adenophorum. Moreover, the translocation of Bax and Cyt c between mitochondria and cytosol triggering the forming of apoptosome proved that the mitochondria-mediated apoptosis was triggered by E. adenophorum. Furthermore, E. adenophorum increased the MDC-positive autophagic vacuoles and the subcellular localization of punctate LC3, the ratio of LC3-II/LC3-I and the protein levels of Beclin 1, but decreased that of P62, indicating the potent pro-autophagic effects of E. adenophorum. In addition, E. adenophorum significantly inhibited the protein leves of p-PI3K, p-Akt and p-mTORC1, but increased PTEN and p-AMPK. Also, the p-mTORC2 and p-Akt Ser473 were inhibited, indicating that the supression of mTORC2/Akt pathway could induce the autophagy of hepatocytes. The autophagy-realted results indicated that the inhibition of PI3K/Akt/mTORC1- and mTORC2/Akt-mediated pathways contributed to the pro-autophagic activity of E. adenophorum. These findings provide new insights to understand the mechanisms involved in E. adenophorum-caused hepatotoxicity of Saanen goat.
... E. adenophorum leaf samples collected from Kangra Valley (India) and partially purified extracts from leaf samples mixed in the diet caused hepatotoxicity and cholestasis in rats 11,12 . Methanolic extract of E. adenophorum leaf samples collected from Mizoram (India) has also been reported to induce hepatotoxicity in albino mice 13 Keeping the above information in view, the present study was undertaken to investigate the status of antioxidant enzymes in mice subjected to oral E. adenophorum extract administration and also to find out whether consumption of the plant is safe or not. ...
Article
Full-text available
Twenty Swiss albino mice (male) randomly divided into four groups were administered orally with vehicle (5% tween 80), 1/20 th (i.e. 175 mg/kg), 1/10 th (i.e. 350 mg/kg) and 1/5 th (i.e. 750 mg/kg) LD 50 doses of methanolic leaf extract of Eupatorium adenophorum respectively; for a period of 30 days and the levels of various antioxidant enzymes such as lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) were studied. Treatment of the mice with methanolic extract of E. adenophorum at the dose levels of 350 and 750 mg/kg revealed marked increase in lipid peroxidation (LPO) levels and decreased activities of superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) as compared to control. The present findings suggest that methanolic leaf extract of E. adenophorum may be having dose dependant hepatotoxic effect on mice as it markedly induced lipid peroxidation and reversed the activities of the antioxidant enzymes.
Article
Full-text available
The cytotoxicity effects of E. adenophorum on cell cycle and apoptosis of renal cells in Saanen goat was evaluated by TUNEL, DAPI, AO/EB staining, DNA fragmentation assay, Caspase activity, Western-blot, qRT-PCR and flow cytometry analysis. 16 saanen goats randomly divided into four groups were fed on 0%, 40%, 60% and 80% E. adenophorum diets. The Results showed that E. adenophorum induced typical apoptotic features of renal cells. E. adenophorum significantly suppressed renal cells viability, caused cell cycle activity arrest and induced typical apoptotic features in a dose-dependent manner. However, the protein levels of Fas/FasL, Bid and caspase-8 did not appear significant changes in the process of E. adenophorum-induced apoptosis. Moreover, E. adenophorum administration slightly decreased Bcl-2 expression, promoted Bax translocation to mitochondria, triggered the release of Cyt c from mitochondria into cytosol and activated caspase-9, -3, and cleaved PARP. The mitochondrial p53 translocation was significantly activated, accompanied by a significant increase in the loss of ΔΨm, Cyt c release and caspase-9 activation. Above all, these data suggest that E. adenophorum induces renal cells apoptosis via the activation of mitochondria-mediated apoptosis pathway in renal cells. These findings may provide new insights to understand the mechanisms involved in E. adenophorum-caused cytotoxicity of renal cells.
Article
Oral administration of the hepatotoxin lantadene A (LA) to guinea pigs elicited a significant rise in the level of bilirubin in blood plasma of the intoxicated animals. Similarly, there were significant increases in the activities of glutamate oxaloacetate transaminase, alkaline phosphatase, acid phosphatase, lactate dehydrogenase, glutamate dehydrogenase, sorbitol dehydrogenase and gamma-glutamyltranspeptidase in the plasma of the intoxicated animals. The alterations in the level of bilirubin and in the blood plasma enzyme profile were consistent with hepatotoxicity and cholestasis. Med Sci Res 27:157-158 (C) 1999 Lippincott Williams & Wilkins.
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A log-probit graph paper is described by which a simple graphic estimation of the ED50 and its standard error may be quickly made.