[Downloaded free from http://www.ijnpnd.com on Friday, November 20, 2015, IP: 112.215.66.

77]

O r i g i n al A r t icle

Toxicological assessment of Pleurotus ostreatus in

Sprague Dawley rats
Krishnamoorthy Deepalakshmi, Sankaran Mirunalini
Department of Biochemistry
and Biotechnology, Faculty of
Science, Annamalai University,
Chidambaram, Tamil Nadu, India
Address for correspondence:
Dr. Sankaran Mirunalini,
Department of Biochemistry and
Biotechnology,
Annamalai University,
Annamalai Nagar,
Chidambaram 608 002,
Tamil Nadu, India.
Email: mirunasankar@gmail.com

ABSTRACT
Objective: To evaluate toxicological and histopathological assessment of Pleurotus
ostreatus, an oyster mushroom in Sprague Dawley rats. Materials and Methods: Toxicity
assessment was carried out by acute (72 h) and sub acute toxicity (28 days) studies
and also its effects on hepatic marker enzymes such as aspartate aminotransferase
(AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and
gammaglutamyl transferase (GGT) and renal markers enzymes (urea, uric acid,
and creatinine) in blood serum and liver histology of experimental Sprague Dawley
rats was performed. Results: The studies indicated that the LD50 value was found
to be >5,000 mg/kg body weight (b.wt). The body weight and general behaviors
of animals were observed throughout the experimental period and at end of the
study, organ weight, biochemical parameters of blood (serum) as well as liver
histology indicated that no toxic clinical symptoms or histopathological changes
were observed in experimental Sprague Dawley rats. Conclusion: The above
studies clearly indicated that the P. ostreatus extract had high margin of safety.

Key words: Acute toxicity, hepatic markers, Pleurotus ostreatus, renal markers, subacute
toxicity

INTRODUCTION
Mushrooms have received a remarkable interest in
recent decades as functional foods and as source
materials for drug development. [1] The major
contributory factors to this growing interest include
rising cost of orthodox medications, low therapeutic
index of synthetic compounds, and the growing
incidence of drug resistance among the pathogens
especially in developing countries with very weak
economic indices.[2] Moreover, the active principles
from natural sources have contributed significantly
to the development of new drugs from herbal
Access this article online
Quick Response Code:
Website:
www.ijnpnd.com
DOI:
10.4103/2231-0738.132665

plants and mushrooms for the treatment of various

diseases.[3,4] Unfortunately, there is limited scientific
evidence regarding safety and efficacy to back up the
continued therapeutic application of these remedies.
The rationale for their utilization has rested largely
on longterm clinical experience. [5] Conversely,
worldwide revolution for the improvement of patient
safety is gaining momentum; hence, the drug safety
for the subject become even more prominent in the
present day scenario.[6] Therefore, closely associated
with screening of plant extracts for their activities
against microorganism or disease conditions is the
need to know their toxic potentials.[7]
Macrofungi have been used as flavorful foods and as
health nutritional supplements since Greek and Roman
days.[8] Basidiomycetes have been widely studied over
the past 30 years in terms of their polysaccharide
composition and therapeutic applications.[9] Among
the various edible species Pleurotus ostreatus is now
rank second among the mushroom consumptioners

International Journal of Nutrition, Pharmacology, Neurological Diseases | July-September 2014 | Vol 4| Issue 3

139

[Downloaded free from http://www.ijnpnd.com on Friday, November 20, 2015, IP: 112.215.66.77]

Deepalakshmi and Mirunalini: Toxicological assessment of Pleurotus ostreatus

in the world.[10] Generally, the P. ostreatus contains

approximately 100 of different bioactive compounds of
each having their own outstanding medical effects.[11]
Furthermore, public is enjoying widespread use of
mushroom for treatment of several ailments, but still
little is known about their toxicity and safety issue
which are always a concern. Hence, evaluation of toxic
properties of a substance is crucial when considering
for public health protection because exposure to
chemicals can be hazardous and results to adverse
effects of human being. The toxicity assessments
includes acute, subacute, and chronic effects.[12]
Thus, the present study aims to determine the
toxicity assessments of ethanolic extract P. ostreatus
using an acute oral toxicity test in animal models.
The acute and subacute toxicity testing was carried
out on animals based on the Organization for
Economic Cooperation and Development (OECD)
guidelines.[13]

MATERIALS AND METHODS

Chemicals

Chemicals and acids were of certified analytical

grade and purchased from S D Fine Chemicals,
Mumbai or HiMedia Laboratories Pvt Ltd, Mumbai,
India.

Material

P. ostreatus mushrooms were collected in and around

areas of Udhagamandalam, Nilgiri district, Tamil
Nadu. The plant was taxonomically identified
and authenticated by Dr V Venkatesalu, Associate
Professor, Department of Botany, Annamalai
University. A voucher specimen (no: 233) was
deposited in the Herbarium of Botany, Department
of Botany, Annamalai University.

Ethanolic extract

The fresh fruiting bodies of P. ostreatus were dried

in shade conditions and the dried materials were
pulverized in a blender to get coarse powder. For
P. ostreatus fruiting bodies ethanolic extraction, 5 g of
the powder was extracted with 100 mL of 95% ethanol
using a Soxhlet apparatus. The solvent was evaporated
under reduced pressure and controlled temperature
(40-50C). The ethanolic extracts were redissolved
in ethanol for the antioxidant activity.[14] A dark,
semisolid material (yield 6 g) obtained was stored at
4C until use. A known amount of the residual extracts
were suspended in distilled water and was orally
administrated to the animals by gastric intubation.
140

Animals and diet

Sixweeksold, female Sprague Dawley rats, weighing

approximately 130 -150 g were obtained from National
Institute of Nutrition, Hyderabad and maintained in
the Central Animal House, Rajah Muthiah Medical
College and Hospital, Annamalai University. All the
rats were acclimatized for a week under standard
husbandry conditions. The rats were housed in
polypropylene cages (45 24 15 cm), maintained
at room temperature (27 2C), in 12 h light/12 h dark
conditions. The animals were fed on a standard pellet
diet (Amrut Laboratory Animal Feed, Mysore Feed
Limited, Bangalore, India) and water ad libitum was
available to the animals throughout the experimental
period and replenished daily. The standard pellet
diet comprised of 21% protein, 5% lipids, 4% crude
fiber, 8% ash, 1% calcium, 0.6% phosphorous, 3.4%
glucose, 2% vitamin, and 55% nitrogen free extract
(carbohydrate) and it provides metabolizable energy
of 3,600 kcal/kg.
Animal handling and experimental procedure
were approved by the Institutional Animal Ethical
Committee of Rajah Muthiah Medical College
(Reg. No: 160/1999, Proposal number: 947 CPCSEA),
and the experiments were performed in accordance
with the Guide for the case and use of laboratory
animal (National Institutes of Health (NIH), 1985) and
committee for the purpose of control and supervision
on experimental animals (Committee for the Purpose
of Control and Supervision of Experiments on Animals
(CPCSEA)).

EXPERIMENTAL DESIGN
Acute toxicity study

The acute toxicity of P. ostreatus was evaluated in

rats using the up and down procedure of OECD
guidelines 423 (adopted December, 2001) with
minor modifications (OECD, 2000). In accordance
with the limit test, single dose of P. ostreatus
(5,000 mg/kg body weight (b.wt)) was orally
administered to three female Sprague Dawley rats
through gastric intubation. The animals were observed
continuously for 72 h for any signs of behavioral
changes and mortality.

Subacute toxicity study

Female Sprague Dawley rates of (130-150 g) were

divided into five groups of six animals each and were
housed under the same conditions as described above.
The ethanolic extract of P. ostreatus was administrated for
28 days at doses of 250, 500, 750, and 1,000 mg/kg b.wt,

International Journal of Nutrition, Pharmacology, Neurological Diseases | July-September 2014 | Vol 4| Issue 3

[Downloaded free from http://www.ijnpnd.com on Friday, November 20, 2015, IP: 112.215.66.77]

Deepalakshmi and Mirunalini: Toxicological assessment of Pleurotus ostreatus

respectively. The control animals received 0.5 mL of

the vehicle alone. Toxic manifestations and mortality
were monitored daily till the end of the experimental
period; all the rats were kept overnight fasting and
anesthetized using ketamine chloride (24 mg/kg b.wt)
by intramuscular injection and sacrificed by cervical
decapitation between 8.00 am to 10.00 am. Blood was
collected in clean dry test tube and serum were used
for various biochemical estimations. Liver tissues were
removed, cleared off blood, and immediately transferred
to icecold containers containing 0.9% NaCl. The tissues
were homogenized in an appropriated buffer and used
for the estimation of various biochemical parameters.

Biochemical estimations

The activity of both serum aspartate aminotranferase

(AST) and alanine aminotransferase (ALT) were
assayed by using the diagnostic kit based on the
method of Reitman and Frankel, 1957.[15] Serum
alkaline phosphatase (ALP) was estimated using
Kind and King, 1954.[16] The serum gammaglutamyl
transferase (GGT) was assayed according to the
method of Rosalki and Rau 1972.[17] Estimation of
renal functional markers such as urea, uric acid, and
creatinine were determined by Fawcett and Scott 1960,
Caraway 1955, and Jeffe 1886.[1820]

Histopathological examination

For histopathological study, three rats from each

group were perfused with physiological saline,
followed by formalin (10% formaldehyde). The liver
tissues were excised immediately and fixed in 10%
formalin. The liver tissues were sliced and embedded
in paraffin wax, 3-5 m thick sections were cut in a
rotary microtome and were stained with hematoxylin
and eosin. The specimens were evaluated with a
light microscope. All histopathological changes were
examined by the pathologist.

Statistical analysis

Statistical analysis was performed using Statistical

Package for Social Sciences (SPSS) software, version
11.5. The values were analyzed by oneway analysis
of variance (ANOVA) followed by Duncans Multiple
Range Test (DMRT). All these results were expressed
as mean standard deviation (SD) for six rats in each
group: P < 0.05 were considered as significant.

RESULTS
Acute toxicity

The body weight and food and water consumption

of rats were found to be unaffected by the treatment
of P. ostreatus extract. No mortality and no significant

changes in general behavior of rats were observed to

the maximum dose level of 5,000 mg/kg b.wt of orally
administered P. ostreatus for 72 h treatment. Hence,
the LD50 was estimated to be > 5,000 mg/kg b.wt.

Subacute toxicity

In order to evaluate the adverse effect of repeated

daily exposure of P. ostreatus, subacute toxicity study
was carried out. To determine doserelated toxic
effects, doses of 250, 500, 750 and 1,000 mg/kg b.wt
of P. ostreatus extract were administered for the
experimental duration of 28 days. P. ostreatus extract
at the different doses did not produce any significant
changes in animals, as evidenced by the absence of
toxic syndromes without any changes in water/food
ingestion and general behaviors.

Effect of P. ostreatus on body weight and organ weight

changes

The body and organ weight of control and

experimental rats were shown in Table1. Oral
administration of P. ostreatus was found to change
the body weights of all the rats, but this was not
statistically significant. The observed weight gain
in the P. ostreatus treated animals shown that
the administrated P. ostreatus does not have any
untoward action that affect the growth of the
animals. Moreover, morphological observation in
vital organ like liver indicated that there was no
sign of any inflammation or toxicity in both control
as well as in the P. ostreatus treated groups. Thus,
the P. ostreatus treated group revealed no significant
difference in body weight or weight of liver organ.
The present result clearly showed that the P. ostreatus
does not produce any toxicological effects on the
body and organ weight.
Table1: Effect of Pleurotus ostreatus on body
weight and liver weight to body weight ratio of
control and experimental animals
Groups

Body weight(g)
Final
Net gain
(%)
Control
157.639.29 180.7011.29 12.76
P. ostreatus 150.568.74 173.499.40
13.22
(250 mg)
P. ostreatus 158.148.52 182.6410.40 13.41
(500 mg)
P. ostreatus 160.238.30 185.5811.71 13.65
(750 mg)
P. ostreatus 159.639.85 184.3011.93 13.38
(1,000 mg)
Initial

Liverwt/
body
wt100

2.320.17
2.460.19
2.490.19
2.500.20
2.560.19

Values are expressed as meanstandard deviation(SD) for six rats.

Comparisons were made between Group I with Groups II, III, IV, and V

International Journal of Nutrition, Pharmacology, Neurological Diseases | July-September 2014 | Vol 4| Issue 3

141

[Downloaded free from http://www.ijnpnd.com on Friday, November 20, 2015, IP: 112.215.66.77]

Deepalakshmi and Mirunalini: Toxicological assessment of Pleurotus ostreatus

Effect of P. ostreatus on biochemical analysis

The activities of hepatic marker enzymes in control and

experimental groups were shown in Table2. However,
no such deleterious changes were found in serum of
liver marker enzymes (ALT, AST, ALP, and GGT) of
P. ostreatus treated groups when compared with the
control rats. The current results clearly indicated that
treatment with P. ostreatus did not induce any harmful
biochemical effects on the animals [Table3] showed
the effect of P. ostreatus on renal functional markers.
The level of serum urea, uric acid, and creatinine
of P. ostreatus treated rats showed no significance
changes like control rats.

Effect of P. ostreatus on histopathology

Histopathological evaluation was carried out to

characterize the biological response factors. Figure1
shows the histopathological examination of liver tissue.
The assessment of histopathology of liver showed
normal architecture implied no detrimental changes
or morphological alterations in control and P. ostreatus
treated animals. This indicates that P. ostreatus did not
exert any toxic effect on the animals.

DISCUSSION
Phytotherapeutic products from medicinal plants have
become universally popular, particularly in developing
countries, and some have been mistakenly regarded as
safe just because they are a natural source. However,
there is a lack of proven scientific studies on the
toxicity and adverse effect of these remedies.[12] An
edible mushroom P. ostreatus has long been known
to be endowed with beneficial and diverse activity.
Therefore, the present study was carried out to
elucidate the acute and subacute toxicity profiles,
including histological evaluation of P. ostreatus in
female Sprague Dawley rats. The toxicity studies are
useful parameter to investigate the therapeutic index
of drug and xenobiotics.[21] At present, the following
chemical labeling and classification of acute systemic
toxicity based on oral LD50 values are recommended by
the OECD (Paris, France): Very toxic, 5 mg/kg; toxic,
>5 and 50 mg/kg; harmful, >50 and 500 mg/kg;
and no label, >500 and 2,000 mg/kg.[22] According to

Table2: Effect of Pleurotus ostreatus on hepatic

marker enzymes in serum of control and
experimental animals
Groups

Control
P. ostreatus
(250 mg)
P. ostreatus
(500 mg)
P. ostreatus
(750 mg)
P. ostreatus
(1,000 mg)

AST
72.846.89
74.125.91

IU/L
ALT
ALP
22.091.84 83.106.19
22.811.94 85.276.53

GGT
2.400.17
2.470.20

75.626.51

23.552.01

86.326.75

2.520.19

77.326.53

24.182.03

87.217.02

2.570.19

79.286.51

24.562.19

88.277.13

2.600.16

AST: Aspartate aminotransferase; ALT: Alanine aminotransferase;

ALP:Alkaline phosphatase; GGT: Gamma glutamyl transferase. Values are
expressed as meanstandard deviation(SD) for six rats. Comparisons were
made between Group I with Groups II, III, IV, and V

Table3: Effect of Pleurotus ostreatus on renal

marker enzymes in serum of control and
experimental animals
Groups

Control
P. ostreatus(250 mg)
P. ostreatus(500 mg)
P. ostreatus(750 mg)
P. ostreatus(1,000 mg)

Urea
25.661.84
26.552.05
27.592.13
28.072.41
28.532.63

mg/dL
Uric acid
1.360.08
1.380.08
1.420.09
1.490.12
1.500.13

Creatine
1.160.08
1.200.10
1.230.09
1.260.08
1.280.09

Values are expressed as meanstandard deviation(SD) for six rats.

Comparisons were made between Group I with Groups II, III, IV, and V

142

Figure1: Histopathological evaluation of liver tissue in control

and experimental animals of Pleurotus ostreatus subacute toxicity
study. (a) Control animals, (b) P. ostreatus treated with 250 mg/kg
body weight (b.wt)), (c) P. ostreatus treated with 500 mg/kg b.wt,
(d) P. ostreatus treated with 750 mg/kg b.wt, (e) P. ostreatus treated
with 1,000 mg/kg b.wt. All groups showing normal lobular architecture
with central vein and radiating hepatic cords. No other histological
changes were noted in all groups of specimens

International Journal of Nutrition, Pharmacology, Neurological Diseases | July-September 2014 | Vol 4| Issue 3

[Downloaded free from http://www.ijnpnd.com on Friday, November 20, 2015, IP: 112.215.66.77]

Deepalakshmi and Mirunalini: Toxicological assessment of Pleurotus ostreatus

the OECD guidelines, the P. ostreatus extract appraisal

no adverse effects were observed in female animals
administrated with up to 5,000 mg/kg b.wt. This
demonstrating the safety median acute toxicity value
(LD50) was estimated to be greater than 5,000 mg/kg
b.wt, respectively.[23] Earlier reports have shown that
if the median lethal dose of a test substance is three
times more than the minimum effective dose, the
substance is considered as good aspirant for the further
studies.[24] Therefore, P. ostreatus extract indicate that it
does not cause any toxicity and safe for oral use for the
management of several diseases.
Nevertheless such acute toxicity data are of limited
clinical application since cumulative toxic effects do
occur even at very low doses. Hence, subacute and
chronic toxicity studies are almost always invaluable in
evaluating the safety profile of phytomedicine.[25] This
probably explains why some authors have suggested
that subchronic toxicity data are inevitable to predict
the hazard of longterm, lowdose exposure to a
particular compound.[26]
In subacute toxicity studies, the body weight and
internal organ weight changes serve as an indicator
of adverse side effects since animals that survive
cannot lose more than 10% of the initial body
weight.[27] In general, toxic nature of the drug leads
to abnormalities in body weight.[28] Nevertheless,
the evaluation of the chronic toxicity at doses of
250, 500, 750, and 1,000 mg/kg b.wt of P. ostreatus
ethanolic extract showed increased in body weight.
Moreover, the increased in body weight was not
significantly different from that of the control. Hence,
we could substantiate that the P. ostreatus ethanolic
extract indicate the improvement in nutritional
state of animals. Earlier reports suggest that the
P. ostreatus had a rich content of protein and the
superior quality of this mushroom may be because
of this genus contain complete proteins with the
well distribution of essential amino acids, as well
as nonessential amino acids, this might be the key
factor for the improved body weight of the rats.[29]
The growth response effect could be a result of
increased food and water intake.[30] Organ weight
also is an important index of physiological and
pathological status in animals. The relative organ
weight was fundamental to diagnose whether the
organ was exposed to the injury or not.[31] However,
the observed results in vital organlike liver indicated
that there were no sign of any inflammation or no
significant differences in organ weight in both control
as well as in P. ostreatus extract treated animals.

Hence, it can be suggested that P. ostreatus ethanolic

extract is virtually nontoxic.
Generally, hepatic cells take part in a variety of
metabolic actions and restrain a host of enzymes.
The biological role of transaminase (AST and ALT),
ALP, and GGT concerned with the interconversion of
highly important metabolite. The enzyme serves as an
index of liver cell injury.[32] However, any elevation
pertaining to these enzymes indicate their outflow into
the blood stream due to damage in liver parenchymal
cells. Thus, liver cell damage is characterized by a rise
in plasma enzymes (AST, ALT, ALP, and GGT).[33] AST
is a liver function test (LFT) and is used to monitor
damage to liver parenchymal cells. Elevated level of
AST is a sign of serious liver damage. ALT is another
enzymes associated with liver parenchymal cells.
ALT is more specific indicator of liver damage than
the AST, as the AST may also be elevated in diseases
affecting other organs. ALT and AST is commonly
measured clinically as a part of diagnostic LFT, to
determine liver health.[34]
ALP is a hydrolase enzyme that eliminated in the bile.
It hydrolyses monophosphate at an alkaline pH. ALP
is particularly present in cells, which line the biliary
ducts of the liver. Generally, heptotoxicity leads to
elevation of normal values due to the bodys inability
to excrete it through bile due to the congestion or
obstruction of the biliary tract.[35] Upgrade in level of
ALP with little or no increase in ALT is primarily a
biomarker of hepatobiliary effects and cholestasis.[36]
GGT or transpeptidase (GGTP) is an enzyme which
is found in liver, kidney, and pancreatic tissues, the
enzyme concentration being low in liver as compared
to kidney. [37] It catalyzes transfer of glutamyl
groups to amino acids and short peptides. It is more
useful clinically when compared to ALP. ALP is
more sensitive, but much less specific than GGT. The
comparison of the two enzymes helps in determining
the occurrence of bone or liver injury. Hence, GGT is
a specific indicator of bile duct lesions in rat liver.[38]
Owing to our results, there were no significant
differences in the serum AST, ALT, ALP, and GGT
levels, which reveal that P. ostreatus did not affect
liver functions or metabolism. It is now established
that excess lipid accumulation in the liver cause
fatty changes and ultimately contains stains such
as lovastatin, which works to reduce cholesterol.
In addition, isolated glucan from P. ostreatus
lowered the serum cholesterol concentration in
hypercholesterolemic rats.[39] Thus, from the above

International Journal of Nutrition, Pharmacology, Neurological Diseases | July-September 2014 | Vol 4| Issue 3

143

[Downloaded free from http://www.ijnpnd.com on Friday, November 20, 2015, IP: 112.215.66.77]

Deepalakshmi and Mirunalini: Toxicological assessment of Pleurotus ostreatus

finding it is evident that usage of P. ostreatus did

not induce any harmful biochemical effects on the
animals.
Kidney is an important organ actively involved in
maintaining homeostasis of the body by reabsorbing
important material and excreting waste products.[40]
Furthermore, kidney functional markers such as
urea, uric acid, and creatinine are the main indicators
of renal dysfunction. Indeed, creatinine is known
as a good indicator of renal function. Any rise in
creatine levels observation is only which marked
damage to functional nephrons.[41] In our study,
there was an insignificant difference in urea, uric
acid, and creatine levels between the treated and
the control group probably indicate that the extract
did not interfere with the renal capacity to excrete
the metabolites. Moreover, the above study is in line
with Jaganathan et al., that the Tridham, a siddha
medicine does not induce any harmful and adverse
effects on the biochemical parameters of Wistar
albino rats.[42]
In general, the histopathology analysis collaborated
with the results of body weight and organ weight.
The P. ostreatus ethanolic extract did not cause toxicity
towards the organs as there was no structural damage
to the organs of liver, kidney, and lungs of the rat. The
liver is the main target organ of acute toxicity, which
was exposed to foreign substances, being absorbed
in intestines, and metabolized to other compounds
which may or may not be hepatotoxic to the rats.[43] In
our study, the liver histopathology analysis showed
the normal hepatocyte architecture and did not cause
any alteration to the structure of the liver cells between
the controls and treated. The above results were in
line with the study by Akanmu et al., 2004 revealed
no necrosis, inflammatory reaction, fibrosis, or local
fatty degradation observed in liver and arrangements
of cell structure almost similar to that of the rats in
control groups.[44] The observation of the current study
portrays that the oral administration of the ethanolic
extract of P. ostreatus did not cause any transience nor
altered the biochemical and histopathological indices
was not harmful at the level tested and can be safely
used as a therapeutic agents.

ACKNOWLEDGMENTS
The authors would like to acknowledge for the Financial Support
from UGC Major Research Project (F. NO: G7/17342/2012 (SR))
to carry out the work successfully.

REFERENCES
1.
2.
3.

4.

5.
6.
7.
8.

9.
10.
11.
12.
13.
14.
15.

CONCLUSION
In conclusion, the present investigation demonstrates
the nontoxic nature of the ethanolic extract of P. ostreatus
was evident from the acute and subacute toxicity
assessments conducted as per OECD guidelines.
Based on 28 days repeated dose toxicity study suggests
144

the P. ostreatus as relatively safe, as it did not cause

either mortality or produce severe toxicological effects
on selected body organs, biochemical indices, and
histological markers of rats. Consequently, P. ostreatus
could be safe up to the dose of 5,000 mg/kg b.wt of the
animals. Owing to this scientific appraisal, it can be
concluded that the ethanolic extracts of the P. ostreatus
had a high margin of safety as it did not induce any
toxicological effects.

16.
17.
18.

Yang JH, Lin HC, Mau JL. Antioxidant properties of several

commercial mushrooms. Food Chem 2002;77:22935.
Seed JR. Current status of African trypanosomiasis. ASM News
2000;66:395402.
Ruknuddin G, Biswajyothi P, Kumar PP, Krishnaiah AB, Basavaiah R.
Antiinflammatory and analagesic activities of Dashanga Ganga: An
ayurvedic compound formulation. Int J Nutr Pharmacol Neurol Dis
2013;3:3038.
Samundeeswari N, Rajadurai M, Ganapathy P, Shoiribha SR. Effecct
of vimliv on lipid profile and histopathology in ethanolinduced
hepatotoxicity in albino wistar rats. Int J Nutr Pharmacol Neurol Dis
2013;3:11420.
Zhu M, Lew KT, Leung PT. Protective effects of plant formula on
ethanolinduced gastric lesions in rats. Phytother Res 2002;16:27680.
Dey YN, Kumari S, Ota S, Srikanth N. Phytopharmacological review of
Andrographis paniculate (Burm. F) wall. Ex Nees. Int J Nutr Pharmacol
Neurol Dis 2013;3:310.
Bulus T, Atawodi SE, Mamman M. Acute toxicity effect of the aqueous
extract of Terminalia avicenniodes on white albino rats. Sci World J
2011;6:14.
Ng TB, Chan WY. Polysaccharopeptides from the mushroom Coriolus
versicolors possesses analgesic activity but does not produce adverse
effects on female reproductive or embryonic development in mice.
Gen Pharmacol 1997;29:26973.
Stamete P. Novel antimicrobials from mushrooms. Herbal Gram
2002;54:2933.
Volz PA. Early historic cancer chemotherapy work involving
basidiomycetous mushrooms. Int J Med Mushrooms 1999;1:1914.
Wang D, Sakoda A, Suzuki M. Biological efficiency and nutritional
values of Pleurotus ostreatus cultivated on spent beer grain. Biosoure
Technolo 2001;78:293300.
Jothy SL, Zakaria Z, Chen Y, Lau YL, Latha LY, Sasidharan S. Acute oral
toxicity of methanolic seed extract of Cassia fistula in mice. Molecules
2011;16:526882.
OECD [Organisation for Economic cooperation and Development]
OECD Guideline for the testing of chemicalsrevised draft guideline
425: Acute oral toxicity: Up and down procedure. Paris: OECD; 2000.
Jayakumar T, Thomas PA, Geraldine P. In vitro antioxidant activities
of an ethanolic extract of the oyster mushroom, Pleurotus ostreatus.
Innov Food Sci Emerg Technol 2009;10:22834.
Reitman S, Frankel S. A calorimetric method for the determination of
serum glutamate oxaloacetic and glutamate pyruvic transaminases.
Am J Clin Pathol 1957;28:5663.
Kind PR, King EJ. Estimation of plasma phosphatases by
determination of hydrolysed phenol with aminoantipyrine. J Clin
Path 1954;7:3226.
Rosalki SB, Rau D. Serum gammaglutamyl transpeptidase activity
in alcoholism. Clin Chim Acta 1972;39:417.
Fawcett JK, Scott JE. A rapid and precise method for the determination
of urea. J Clin Path 1960;3:1569.

International Journal of Nutrition, Pharmacology, Neurological Diseases | July-September 2014 | Vol 4| Issue 3

[Downloaded free from http://www.ijnpnd.com on Friday, November 20, 2015, IP: 112.215.66.77]

Deepalakshmi and Mirunalini: Toxicological assessment of Pleurotus ostreatus

19. Caraway WT. Determination of uric acid in serum by carborated
method. Am J Clin Pathol 1955;25:8405.
20. Jeffe M. Concerning the precipitate produced in normal urine by picric
acid and a new reaction of creatinine. Physiol Chem 1886;10:91400.
21. Rang HP, Dale M, Raitter J. Pharmacology. Vol. 13, 4th ed. New York:
Churchill Livingstone, USA; 2001.
22. Walum E. Acute oral toxicity. Environ Health Perspect 1998;106:497502.
23. Ghosh MN. In statistical analysis, Fundamentals of experimental
pharmacology, 2nd ed. Calcutta: Scientific Book Agency; 1984. p. 187-9.
24. Salawu OA, Chindo BA, Tijani AY, Obidike IC, Salawu TA, James
Akingbasote A. Acute and subacute toxicological evaluation of the
methanolic stem bark extract of Crossopteryx febrifuga in rats. Afr J
Pharm Pharmacol 2009;3:6216.
25. Aniagu SO, Nwinyi FC, Akumka DD, Ajoku GA. Toxicity studies in
rats fed nature cure bitters. Afr J Biotechnol 2005;4:728.
26. Mc Namara BP. Concept in health evaluation of commercial and
industrial chemicals. In: Mehlman MA. Shapiro RE, Blumental H,
editors. New Concepts in Safely Evaluation. Washington: Hemispher;
1976.
27. Raza M, Alshabanah OA, ElHadiyah TM, AlMajed AA. Effects of
prolonged vigabatrin treatment on haematological and biochemical
parameters in plasma, liver and kidney of swiss albino mice. Sci
Pharma 2002;701:13545.
28. Etuk EU, Muhammad AA. Safety evaluations of aqueous stem bark
extract of Lophira lanceolata in Sprague dawley rats. Int J Res Pharm Sci
2010;1:2833.
29. Akyuz M, Kirbag S. Nutritive values of wild edible and cultivated
mushrooms. Turk J Biol 2010;34:97102.
30. Uma M, Suresh M, Thulasiraman K, Lakshmidevi E, Kalaiselvi P.
Chronic toxicity studies of aqueous leaf extract of Indian traditional
medicinal plant Ocimum tenuiflorum (Linn.) in rats. Eur J Exp Biol
2013;3:2407.
31. Dybing E, Doe J, Groten J, Kleiner J, OBrien J, Renwick AG, et al.
Hazard characterization of chemicals in food and diet. Dose response,
mechanism and extrapolation issue. Food Chem Toxicol
2002;42:23782.
32. Hanley AJ, Williams K, Festa A, Wagenknecht LE, DAgrostino RB Jr,
Haffner SM. Liver marker and development of the metabolic syndrome:
The insulin resistance atherosclerosis study. Diabetes 2005;54:31407.
33. Mayne PD. Clinical chemistry in Diagnosis and treatment, 6th ed.
(International students Edition). New York: Arnold London/Oxford
University Press Inc; 1996.
34. Alam N, Hossain MS, Khair A, Amin SM, Khan A. Comparative effects

35.

36.
37.
38.
39.

40.
41.
42.

43.
44.

of mushroom on plasma lipid profile of hypercholestrolaemic rats.

Bangladesh J Mushroom 2007;1:1522.
Saukkonen JJ, Cohn DL, Jasmer RM, Schenker S, Jereb JA,
Nolan CM, et al. ATS (American Thoracic Society) Hepatotoxicity of
Antituberculosis Therapy Subcommittee. An official ATS statement:
Hepatotoxicity of antituberculosis therapy. Am J Respir Crit Care
Med 2006;174:93552.
Ramaiah SK. A toxicologist guied to the diagnostic interpretation of
hepatic biochemical parameters. Food Chem Toxicol 2007;45:15517.
Ozer J, Rather M, Shaw M, Bailey W, Schomaker S. The current state
of serum biomarkers of hepatotoxicity. Toxicology 2008;245:194205.
Leonard TB, Neptun DA, Popp JA. Serum gamma glutamyl transferase
as a specific indicator of bile duct lesions in the rat liver. Am J Pathol
1984;116:2629.
Hossain S, Hashimoto M, Choudhury EK, Alam N, Hussain S,
Hasan M, et al. Dietary mushroom (Pleurotus ostreatus) ameliorates
atherogenic lipid in hyper cholesterolaemic rats. Clin Exp Pharmacol
Physiol 2003;30:4705.
Arulmozhi V, Krishnaveni M, Mirunalini S. Protective effect of Solanum
nigrum fruit extract on the functional status of liver and kidney against
ethanol induced toxicity. J Biochem Tech 2012;3:33943.
Mukinda JT, Eagles PF. Acute and subchronic oral toxicity profile
of the aqueous extract of Polygala fruticose in female mice and rats. J
Ethanopharmacol 2010;128:23640.
Jaganathan R, Ravinayagam V, Panchanadham S, Palanivelu S.
Toxicological, biochemical and histopatological evaluation of
tridham, a siddha medicine in wistar albino rats. J Biochem Tech
2012;4:5418.
Rhiovania H, ElHilalya J, Israili ZH, Lyoussia B. Acute and subacute
toxicity of an aqueous extract of the leaves of Herniaria glabra in
rodents. J Ethnopharmacol 2008;118:37886.
Akanmu MA, Iwalewa EO, Elujoba AA, Adelusola KA. Toxicity
potentials of Cassia fistula fruits as laxative with reference to senna.
Afr J Biomed Res 2004;7:236.

How to cite this article: Deepalakshmi K, Mirunalini S. Toxicological

assessment of Pleurotus ostreatus in Sprague Dawley rats. Int J Nutr
Pharmacol Neurol Dis 2014;4:139-45.
Source of Support: UGC Major Research Project. Conflict of
Interest: None declared.
Received: 03-03-2014, Accepted: 23-03-2014

Author Help: Online submission of the manuscripts

Articles can be submitted online from http://www.journalonweb.com. For online submission, the articles should be prepared in two files (first
page file and article file). Images should be submitted separately.
1) First Page File:

Prepare the title page, covering letter, acknowledgement etc. using a word processor program. All information related to your identity should
be included here. Use text/rtf/doc/pdf files. Do not zip the files.
2) Article File:

The main text of the article, beginning with the Abstract to References (including tables) should be in this file. Do not include any information
(such as acknowledgement, your names in page headers etc.) in this file. Use text/rtf/doc/pdf files. Do not zip the files. Limit the file size
to 1024 kb. Do not incorporate images in the file. If file size is large, graphs can be submitted separately as images, without their being
incorporated in the article file. This will reduce the size of the file.
3) Images:

Submit good quality color images. Each image should be less than 4096 kb (4 MB) in size. The size of the image can be reduced by decreasing
the actual height and width of the images (keep up to about 6 inches and up to about 1800 x 1200 pixels). JPEG is the most suitable file
format. The image quality should be good enough to judge the scientific value of the image. For the purpose of printing, always retain a good
quality, high resolution image. This high resolution image should be sent to the editorial office at the time of sending a revised article.
4) Legends:

Legends for the figures/images should be included at the end of the article file.

International Journal of Nutrition, Pharmacology, Neurological Diseases | July-September 2014 | Vol 4| Issue 3

145