American Chemical Science Journal

4(1): 74-81, 2014

SCIENCEDOMAIN international
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Effects of Aqueous Extract of Ocimum

gratissimum on Some Haematological
Parameters of Albino Rats
O. A. Ojo1*, O. I. Oloyede2, B. O. Ajiboye1 and O. I. Olarewaju2
1

Department of Chemical Sciences, Biochemistry Unit, Afe Babalola University, Nigeria.

2
Department of biochemistry, Ekiti State University, Ado-Ekiti, Nigeria.
Authors contributions

Author OAO carried out the experiment and performed the statistical analysis, author
Oloyede designed the study, author BOA managed the analyses of the study, and author
Olarewaju managed the literature searches. All authors read and approved the final
manuscript.

th

Original Research Article

Received 28 May 2013

st
Accepted 21 August 2013
st
Published 21 October 2013

ABSTRACT
Aims: To evaluate the effects of aqueous extracts of Ocimum gratissimum leaves on the
haematological responses of albino rats.
Study Design: Packed cell volume, hemoglobin level, white blood cells and differential cell
counts such as neutrophils, lymphocytes, monocytes and eosinophils.
Place and Duration of Study: Department of Chemical sciences, (Biochemistry
laboratory), Afe Babalola University and Department of Biochemistry, Ekiti State University,
Ado Ekiti, Nigeria between February 2013 and march 2013.
Methodology: Sixteen albino rats with weight ranging between 100-190g were used. The
rats were divided into four groups A, B, C and D; with group D as the control group.
-1
Increasing doses (100, 200 and 400mg kg body weight) of the extract were administered
orally to the other three treatment group A, B and C for a period of two weeks. Standard
chemical procedures were used to analyze for packed cell volume, hemoglobin level, white
blood cells and differential cell counts such as neutrophils, lymphocytes, monocytes and
eosinophils.
Results: The results show a significantly decrease in the hemoglobin level, packed cell
volume, white blood cells, neutrophils and lymphocytes respectively. Significant (P = .05)
____________________________________________________________________________________________
*Corresponding author: Email: oluwafemiadeleke08@gmail.com;

American Chemical Science Journal, 4(1): 74-81, 2014

decreases in the level of hemoglobin (Hb) and packed cell volume were observed,
whereas no significant (P > .05) differences were observed in percent neutrophils,
lymphocyte and monocyte counts of the Ocimum gratissimum treated animals.
Conclusion: The result suggests that Ocimum gratissimum leaf extracts administered at
the dosages used and for the duration of the experiment suppress the haemopoetic
system of albino rats.

Keywords: Aqueous extract; Ocimum gratissimum; haematological parameters; albino rats.

1. INTRODUCTION
Ocimum gratissimum is a perennial herb belonging to the family Lamiaceae. The plant which
is believed to originate from Asia and Africa [1] is also widely distributed in other regions,
occurring as different species. The leaves of the plant contain essential oils [2] and it is
cultivated for various purposes. In Nigeria and most parts of West Africa, it is used as a
spice and condiment in dishes, because of its high pungent flavor of clove. The whole plant
and the essential oils of the plant have many applications in folk medicine. The crude
extracts have been used in the treatment of respiratory tract infections [3]; diarrhea [4];
rheumatism and hemorrhoids [1,5]; dermatological conditions [6,7]; microbial infections [8];
epilepsy, high fever and mental illness [9,10]. Phytochemical screening of the leaf extract of
Ocimum gratissimum (OG) had shown the plant to contain alkaloids, saponins, tannins,
alkaloids, anthraquinone, flavonoids, steroids, terpenoids and cardiac glycosides [11,8,12].
Furthermore, OG had been shown to possess diverse pharmacological properties which
may be attributed to its usefulness in folk medicine. These properties include antioxidant
[12,13,14] chemotherapeutic [15]; antimutagenic [16]; antidiarrhoeal [6,17,18] antinociceptive
[19] insecticidal [20] hypotensive [21] and antihelmintic [22,23]. The chemical composition of
Ocimum gratissimum are pinene, -pinene, 1,8-cineole,Terpineole, Eugenol, (E)-Caryoplyllene, Muurolere, Sehirene and -selinerel [24]. Erythrocyte (red blood cell) which
are anucleate are packed with the oxygen carrying protein hemoglobin. Human erythrocytes
survive in the circulation for about 120 days, worn out erythrocytes are removed from the
circulation by macrophages of the spleen and bone marrows. The signal for removal seems
to be the appearance of defective complex oligosaccharides attached to integral membrane
protein of the plasmalemma [25]. Due to the widespread consumption of Ocimum
gratissimum, it is necessary to study its effect on blood, the tissue that transports substances
in the body [25]. This study was therefore designed to evaluate the effects of aqueous
extracts of O. gratissimum leaves on the haematological responses of albino rats.

2. MATERIALS AND METHODS

2.1 Plant Material
Fresh leaves of O. gratissimum were bought in the market at Ado, Nigeria. The plant was
identified and authenticated by a plant scientist in the Department of Plant Science, Ekiti
State University, Ado-Ekiti, Nigeria and a voucher specimen was deposited accordingly at
the herbarium of the Department of Plant Science, Ekiti State University, Ado-Ekiti, Nigeria.

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American Chemical Science Journal, 4(1): 74-81, 2014

2.1.1 Plant extraction

The fresh leaves of the plant were air-dried, pulverized and extracted exhaustively in distilled
water. The filtrate was concentrated and evaporated to dryness at 60C, using rotary
evaporator. The yield was calculated and the dry extract was stored in a refrigerator at -4C
until use for the experiments.

2.2 Animals
A total number of 16 inbred albino male rats weighing between 100-190 g were used in this
study. The animals were obtained from the animal house of the Department of Chemical
Sciences, Afe Babalola University, Ado-Ekiti, Nigeria. The animals were randomly distributed
into cages and allowed to acclimatize for 7 days in a well ventilated room at a room
temperature of 20.02.0C under natural lighting co ndition. The animals were allowed free
access to standard rat chow (Topfeeds Ltd., Ado-Ekiti, Nigeria) and distilled water ad libitum.
All animals used in this study were handled in accordance with the international, national
and institutional guidelines for Care and Use of Laboratory Animals as promulgated by the
[26].

2.3 Experimental Protocol

Animals were divided into four groups- A, B, C and D (control) with four experimental
animals in each group, respectively and were kept in a metabolic cage. Group A, B and C
-1
-1
-1
received daily doses of 100 mg kg , 200 mg kg and 400 mg kg of OG for 14 days. The
control group (group D), containing four animals, was given only distilled water daily for 14
days. OG was administered orally using a calibrated 1 mL syringe with attached polythene
cannula. There were changes in the physical appearance of the animals as they increase in
body weight.

2.4 Haematological Assays

Blood samples were collected from the animals through venipuncture and analyzed using
standard methods for packed cell volume (PCV), hemoglobin (Hb), WBC count and WBC
percentage composition using standard techniques. Total white blood cell counts (WBC)
were determined by the haemocytometer method, while the smears were prepared and
stained by the Leishman technique and enumerated by the longitudinal counting method to
determine differential count [27]. The packed cell volume (PCV) was determined by the
microhaematocrit method while haemoglobin (Hb) concentration was determined by the
cyanomethaemoglobin method [27,28].

2.5 Statistical Analysis

Data were expressed as Mean SE of mean. Comparisons between control values and
values of treated groups of albino rats were performed with one-way Analysis of Variance
(ANOVA) with the help of software SPSS 16.0 for windows. Statistical significance was set
at p = .05. Statistical significance was tested using students t-test, compared with control
value P = 0.05.

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3. RESULTS AND DISCUSSION

3.1 Haematological Parameters
The results of the effect of different doses of aqueous extract of Ocimum gratissimum on the
haematological parameters are shown in Table 1.
Table 1. Some haematological parameters as well as WBC count and WBC
composition in the tested animal groups
Parameters
Hb (g/dl)
PCV (%)
3
3
WBC (x 10 mm )
Neutrophils (%)
Lymphocytes (%)
Eosinophils (%)
Monocytes (%)
abc

Group A
ab
14.15 4.79
b
31.25 1.80
a
3.45 64.55
a
49.50 2.53
a
42.00 3.25
a
1.75 0.48
a
6.75 1.08

Group B
a
13.95 7.93
a
26.75 0.85
b
4.78 85.39
a
46.00 1.29
a
43.50 1.05
a
2.75 0.48
a
7.75 0.63

Group C
a
14.05 2.22
c
37.50 1.04
a
3.78 2.10
a
47.25 0.63
a
42.50 1.55
a
3.25 0.85
a
7.00 1.65

Group D
b
15.65 2.5
c
35.50 0.65
b
4.78 85.39
a
45.50 1.32
a
44.50 1.44
a
2.75 0.48
a
7.25 0.91

Means within a row with different superscripts are significantly (P = .05) different

This work tested the effect of aqueous extract of Ocimum gratissimum on Haematological
parameters of albino rats. The phytochemical constituents of the aqueous leaf extract of
Ocimum gratissimum contains Saponins, flavonoids, alkaloids, phenol, terpenes and
steroids, tannins and cardiac glycosides as reported by [29]. Almost all the photo
constituents of Ocimum gratissimum are known to influence biological system activities. The
results of the study show that, the leaf extract of Ocimum gratissimum administered at the
dosages used and for the duration of the experiment suppress the haemopoetic
system. The hemoglobin values at the end of the experiment were 15.65 2.5 for the
control group D and 14.15 4.79, 13.95 7.93, and 14.05 2.22 for the experimental
groups. This shows a reduction in the hemoglobin level, this is in consonance with the
finding of [30]. The haemoglobin (Hb) levels of the control group D animals (15.65 2.5)
were significantly (P = .05) higher than treatment group B (13.95 7.93) and group C (14.05
2.22), which were similar (P > .05) but no significant difference to group A (14.15 4.79).
The packed cell volume of the control group D (35.50 0.65) were significantly (P = .05)
higher than treatment group A (31.25 1.80) and group B (26.75 0.85) but similar (P > .05)
to group C (37.50 1.04). Treatment group A (31.25 1.80) is also significantly (P = .05)
higher than group B (26.75 0.85). The white blood cell count gives a mean value of (4.78
3
3
85.39) (x 10 mm ) for the control group D animals and were significantly (P = .05) higher
3
3
3
3
than treatment group A (3.45 64.55) (x 10 mm ) and group C (3.78 2.10) (x 10 mm ) but
3
3
no significant difference (P > .05) to group B (4.78 85.39) (x 10 mm ), while treatment
3
3
3
3
group A (3.45 64.55) (x 10 mm ) and group C (3.78 2.10) (x 10 mm ) were not
significantly different (P > .05). Percentage neutrophils counts of the animals for control
group D (45.50 1.32), group A (49.50 2.53), group B (46.00 1.29) and group C (47.25
0.63) were not significantly different (P > .05). The percentage lymphocytes of the animals
for control group D (44.50 1.44 %), group A (42.00 3.25), group B (43.50 1.05) and
group C (42.50 1.55) were not significantly different (P > .05). The lymphocyte level
decreased at the end of the experiment as compared to the control group D. This is
supported by the findings of Ephraim et al., 2000. The percentage eosinophils counts of the
animals for control group D (2.75 0.48), group A (1.75 0.48), group B (2.75 0.48) and
group C (3.75 0.48) were not significantly different (P > .05). Percentage monocytes for
control group D (7.25 0.91), group A (6.75 1.08), group B (7.75 0.63) and group C

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American Chemical Science Journal, 4(1): 74-81, 2014

(7.00 1.65) were not significantly different (P > .05). In addition, 2 weeks administration of
OG results in no significant differences in lymphocyte, eosinophils and monocytes counts
between the experimental groups and control group. Earlier studies had shown that the
aqueous leaf extract of O. gratissimum has different pharmacological actions including
antioxidative properties [19; 13; 2; 14]. Obianime et al. [31] reported decreased Hb and PCV
with O. gratissimum leaf extracts in haematological analysis of male mice. Furthermore, the
results of this study show that the hematopoietic system is sensitive to O. gratissimum
causing anemia and increase in leucocytes during the administration (2 weeks). These
effects may be due to oxidative compounds like saponnins and triterpenes which are
contained in the crude extract of O. gratissimum which are deleterious to blood cells [32].
The reduced hematological effects of O. gratissimum after administration in the present
study may be due to adaptation through stimulation of adaptive mechanisms by the body
against O. gratissimum-induced toxicity on the blood cells. It thus suggests that O.
gratissimum may have dual responses (i.e., oxidative or antioxidative), depending on the
tissue/organ system under investigation or the duration of administration. This may be
explained by the complex amount or nature of phytochemical compounds contained in the
crude extract of O. gratissimum which have both oxidative (e.g., saponnins, triterpenes,
alkaloids) and antioxidative (e.g., eugenol, flavonoids, citral, linalool) properties. This
decrease could be as a result of genetic differences the in animals of their experiment and
the ones used in this study. This result suggests that O. gratissimum leaf extracts
administered at the dosages used and for the duration of the experiment suppress the
haemopoetic system [32].

4. CONCLUSION
In conclusion, the study shows that the leaf extract of Ocimum gratissimum administered at
the dosages used and for the duration of the experiment suppress the haemopoetic system.
The reduction may have occurred due to lysis of blood cells and probably suppression of
blood cell synthesis by saponins found in the leaf extract [32]. Saponins are known to be
toxic to body systems [33]. Despite of the popularity of the plant as a condiment and herbal
medicine, the extract has been observed to suppress the haemopoetic system. It is therefore
suggested that chronic usage of the leaf is not advisable.

ACKNOWLEDGEMENTS
The Authors wish to acknowledge Mr. Omotayo of the Department of Plant Science, Ekiti
State University, Ado-Ekiti, Nigeria in helping to identify the plant and providing the voucher
number.

ETHICAL APPROVAL
All authors hereby declare the international, national and institutional guidelines for care and
use of laboratory animals as promulgated by the Canadian council on animal care, 2009, the
care and use of farm animals in research, teaching and testing. CCAC. OTTAWA, pp: 12-15
were followed, as well as specific national laws where applicable. All experiments have been
examined and approved by the appropriate ethics committee.

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COMPETING INTERESTS
Authors have declared that no competing interests exist.

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2014 Ojo et al.; This is an Open Access article distributed under the terms of the Creative Commons Attribution
License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction
in any medium, provided the original work is properly cited.

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