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Immuno Modulatory Effects of Hot Aqueous Extract of Ocimum sanctum and Argemone mexicana Leaves in Chicken Model

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The present study was undertaken to study the immuno modulatrory effects of leaves of Ocimum santum (OS) and Argemone mexiana (AM) plants in chicken model. 250 mg/kg body weight oral dose of OS and AM was found ideal and nontoxic in chickens and experimental chickens were fed this dose. Humoral immune response to S. enterica serovar Typhimurium 'O' antigen was measured by quantitating the serum antibody level of immunized chickens by ELISA test. There was a significant rise in antibody titre of OS and AM fed chickens in comparison to control group. The antibody titre in the serum samples of HAE of OS and AM fed chickens were (5333.33±674.62) and (4266.67±674.62) respectively, whereas in control group it was 3733.33±533.33. This study indicated that the extract of both plants enhanced the antibody level and acted as a humoral immuno stimulant. Cell mediated immune response (CMI) was assayed by differential lymphocyte count (DLC) and DNCB sensitized hypersensitivity test. DLC revealed increase in lymphocyte count in OS fed group and decrease in lymphocyte count in AM fed group as compared to control group. DNCB test demonstrated 29.89% increase in skin thickness in OS fed group and 34.02% decrease in skin thickness in AM fed group as compared to control group at 24 hour interval. Present study indicated the T cell suppressive impact of hot aqueous extract of AM and stimulator effect of OS.
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Research Report Open Access
Immuno Modulatory Effects of Hot Aqueous Extract of Ocimum sanctum and
Argemone mexicana Leaves in Chicken Model
Puneet Varshney Sandeep Dash Anjana Goel Ashok Bhatia
Department of Microbiology and Immunology, College of Veterinary Science and Animal Husbandry, DUVASU, Mathura - 281001, UP, India
Corresponding author email: sandeepkumar.dash@gmail.com; Authors
Medicinal Plant Research, 2013, Vol.3, No.8 doi: 10.5376/mpr.2013.03.0008
Received: 15 May, 2013
Accepted: 23 May, 2013
Published: 27 May, 2013
Copyright © 2013 Varshney et al. This is an open access article published under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Varshney et al., 2013, Immuno Modulat Effects of Hot Aqueous Extract of Ocimum sanctum and Argemone mexicana Leaves in Chicken Model, Medicinal
Plant Research, Vol.3, No.8 57-62 (doi: 10.5376/mpr.2013.03.0008)
Abstract The present study was undertaken to study the immuno modulatrory effects of leaves of Ocimum santum (OS) and
Argemone mexiana (AM) plants in chicken model. 250 mg/kg body weight oral dose of OS and AM was found ideal and nontoxic in
chickens and experimental chickens were fed this dose. Humoral immune response to S. enterica serovar Typhimurium ‘O’ antigen
was measured by quantitating the serum antibody level of immunized chickens by ELISA test. There was a significant rise in
antibody titre of OS and AM fed chickens in comparison to control group. The antibody titre in the serum samples of HAE of OS and
AM fed chickens were (5333.33±674.62) and (4266.67±674.62) respectively, whereas in control group it was 3733.33±533.33. This
study indicated that the extract of both plants enhanced the antibody level and acted as a humoral immuno stimulant. Cell mediated
immune response (CMI) was assayed by differential lymphocyte count (DLC) and DNCB sensitized hypersensitivity test. DLC
revealed increase in lymphocyte count in OS fed group and decrease in lymphocyte count in AM fed group as compared to control
group. DNCB test demonstrated 29.89% increase in skin thickness in OS fed group and 34.02% decrease in skin thickness in AM fed
group as compared to control group at 24 hour interval. Present study indicated the T cell suppressive impact of hot aqueous extract
of AM and stimulator effect of OS.
Keywords Argemone mexicana; Ocimum sanctum; Hot aqueous extract; Humoral; CMI; DNCB
Background
There are several diseases which are the result of
immunosuppression of either humoral or cell mediated
or both type of responses. In such cases often chemical
drugs fail to restore the normal functions. Under such
circumstances modulation of immune system by
medicinal plants or their products could be a
possible alternative therapeutic approach. World health
organization (WHO) has started to give emphasis on
development and use of herbal products for the benefit of
world population in viewing the limitations and ill effects
of chemical drugs (Lee, 2004). The ecofriendly and
non-hazardous nature of herbal medicines for humans
and animals, absence of residual effects, minimum
problems of drug resistance and absence of side effects
further instill the interest in herbal medications.
Among these plants O. sanctum occupies significant
place in the indigenous system of medicine of many
Asian, African and South American countries. In India
the practitioner of traditional system of medicine have
used O. sanctum for curing various ailments. From
last two decades, to prove the scientific basis of
therapeutic use of O. sanctum in modern medicine,
several researchers (Sood et al., 2006; Bhartiya et al.,
2006; Goel et al., 2010) have begun the exploring the
pharmacological effects of this plant and observed that
these extracts had modulation effect on the immune
responses by regulating cytokines.
Besides these well-established medicinal herbs, there
are some weeds like A. mexicana known for their
toxicity may have biological activities beneficial to
animal kingdom including birds (Kumar, 2006). A.
mexicana could be attributed for various disorders
scabies, pruritis, eczema and psoriasis, a disease of
impaired or defective cell mediated immunity and
used in treatment of cataracts and opthalmia (Oudhia,
2001, Gupta, 2004, Kumar, 2006, Goel et al., 2008).
1 Results
1.1 Determination of nontoxic dose
It is well known that seeds of A. mexicana, contain
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toxic substance called sanguinarine which on
consumption produces dropsy in human and animals.
However in our study aqueous extract was prepared
from A. mexicana leaves, GC-MS & HPTLC of A.
mexicana leaves powder revealed the absence of
sanguinarine compound. All the three doses of OS and
AM were safe and nontoxic as no untoward clinical
sign was observed in any of the birds. All the birds
were apparently normal and healthy. No adverse effect
on hematological parameters was found (data not
shown). For humoral and CMI response study OS/AM
dose of 250 mg/kg was used as it was ideal.
1.2 Humoral immune responses against S. enterica
serovar Typhimurium “O” antigen
Antibody titre in chickens of experimental and control
groups against S. enterica serovar Typhimurium ‘O’
antigen was estimated by indirect ELISA. Before
immunization the antibody titer in all the three groups
was similar ranging from 933.33±133.33 to
933.33±458.12. 7th day after 1st immunization blood
sera collected from O. sanctum and A. mexicana
extract showed rise in antibody titer to 2800.00±400
and 2266.67±434.1 respectively. The antibody titer
further rose to 5333.33±674.62 and 4266.67±674.62
respectively in O. sanctum and A. mexicana fed
groups when measured at 7th day after 2nd
immunization. The rise in antibody titer was
significant in O. sanctum and A. mexicana chickens as
compared to control group. The mean values of
antibody titer are given in Table 1 and Figure 1.
Table 1 Humoral immune response in chicken against S. enterica serovar Typhimurium “O” antigen
Antibody titer
Control group O. sanctum fed group
A
. mexicana fed group
At 0 day (pre vaccination) 933.33 ±133.33 933.33±458.12 933.33±223.11
After 7 days of vaccination 2133.33±337.31 2800.00±400.00 2266.67±434.10
After 14 days of vaccination 3733.33±533.33 5333.33±674.62 4266.67±674.62
Note: All values are mean ± S.E. of 6 birds
Figure 1 Line diagram showing Humoral Immune response
using S. enterica serovar Typhimurium “O” antigen in fed and
unfed groups
1.3 Cell mediated immune (CMI) response in
chickens using DNCB
For studying the effect of HAE of OS and AM leaves
on Cell mediated immune response, in vivo assay was
employed to assess the delayed type hypersensitivity
(DTH) response in chickens using DNCB as an
allergen. Mean values of skin thickness of chickens of
experimental (OS and AM fed) and control groups at
24 h, 48 h and 72 h post challenge of DNCB are
presented in Table 2 and Figure 2. In relation to
control group, skin thickness increased markedly in
OS fed chickens which were 29.89% higher at 24 h. In
AM fed groups thickness was lower than that of
control group showing 34.02% inhibition at 24 h. Skin
thickness when measured at 48 h and 72 h, OS fed
chickens group remained higher 22.98% and 8.10 %
increase in comparison to control. In case of AM fed
group there was overall inhibition of skin thickness.
HAE of OS showed marked cell mediated immune
response but AM showed suppression of cell mediated
immune response.
2 Discussion
Immunomodulatory responses of HAE of OS and AM
leaves were evaluated by determining the humoral and
cell mediated immune responses. Many workers
(Godhwani et al., 1988 and Mediratta et al., 2002) also
studied OS effect on immunocytes and found it
immunocyte stimulator. Babu et al., (2001) reported
that immunomodulatory action is mostly concerned
with cellular involvement of haemopoetic and lymphoid
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Table 2 Mean values of skin thickness in chickens of control, OS and AM fed groups at different time intervals
Group Mean thickness
(mm) at 0 h
Mean thickness
(mm)
Difference in thickn-
ess (mm)
Percent change in
thickness
Percent change comp-
ared to control
24 h 48 h 72 h 24 h 48 h 72 h 24 h 48 h 72 h 24 h 48 h 72 h
Control 1.25 2.22 2.12 1.62 0.97 0.87 0.37 77.6 69.6 29.6 - - -
OS 1.33 2.59 2.40 1.73 1.26 1.07 0.4 94.73 80.45 30.07 29.89 22.98 8.1
AM 1.30 1.94 2.05 1.66 0.64 0.75 0.36 49.23 57.69 27.69 -34.02 -13.79 -2.7
Figure 2 Line diagram showing difference in skin thickness
(mm) in OS and AM fed groups
tissues. Humoral immune response was quantitated by
measuring antibodies by indirect ELISA using S.
enterica serovar Typhimurium ‘O’ antigen and results
were compared with control (unfed chickens). It was
observed that antibody titre in the serum samples of
HAE of OS and AM fed chickens were higher
(5333.33±674.62) and (4266.67±674.62) respectively,
whereas unfed control group exhibited 3733.33±533.33
antibody titre (Table 1 and Figure 1). This indicates
that the HAE of OS and AM enhanced the antibody
level 42.85% and 14.28% respectively in comparison
to control group and acted as humoral response
stimulant. These results were reproducible and
consistent. Earlier Kumar (2006) also observed similar
results using S. enterica serovar Typhimurium antigen
in rabbits. Sadekar et al (1998) also demonstrated
enhance antibody response in OS leaves fed birds
having infection of IBD virus. Maims (2004) reported
effective role of OS in enhancing the immune system
against bacterial and viral infections. Lombal et al
(2004) observed the specific and nonspecific immune
responses on account of OS leaves extract and found it
to stimulate antibody response. Our results are similar
to their findings.
To find the effect of HAE of these plants leaves on
cell mediated immune response skin hypersensitivity
reaction was done using DNCB as an allergen. In
Ocimum sanctum treated group there was 94.73%
increase in skin thickness whereas in Argemone
mexicana treated group 49.23% increase compared to
control group which showed 77.6% increase at 24 hrs.
The analysis revealed increase (29.82%, 22.98% and
8.10%) in OS fed group while as decrease (-34.02%,
-13.79% and -2.70%) in AM fed chickens as
compared to control group after 24, 48 and 72 hrs
respectively (Table 2 and Figure 2). Overall results
obtained in the present study showed that HAE of OS
has stimulatory influence and AM has inhibitory effect
on CMI in chickens. This study indicated the T cell
suppressive impact of aqueous extract of Argemone
mexicana. Recently Goel et al (2010) observed a
stimulatory effect of ocimum sanctum on humoral and
cell mediated immune response. Humoral immune
response observed by increased antibody titre against
S. enterica serovar Typhimurium O’ antigen and
significant increase in skin thickness by DNCB
hypersensitivity test. The stimulant effect of Argemone
mexicana on humoral immune response and inhibitory
effect on CMI is also reported by Goel et al (2008).
Mediratta et al (2002) investigated the effect of OS
seed oil on stressed and non-stressed animals and
found that OS modulated both humoral and cell
mediated immune response and suggested that this
may be due to mediation of GABAergic pathway. In
our study HAE of OS was found to have stimulatory
effect on both arms of immune system while HAE of
AM exhibited antibody stimulation response but
caused suppression in CMI response. This study
indicates that OS can be used against bacterial, viral
infection as well as chicken having immuno
suppression. While HAE of AM can be used to treat
diseases due to unusual elevated cell mediated
immune response (Gupta, 2004; Kumar, 2006; Goel
et al., 2008). Mukherjee et al (2005) reported that
OS leaves possess some biologically active
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immunostimulatory components. Goel et al (2010)
also found the similar results of Ocimum sanctum as,
antiviral and immunomodulator plant on rat, besides
reported the increased level of interferon gamma
cytokine after orally fed of OS to the experimental rat.
This study conclude that at the present time when we
are facing with the limitations of modern medical
science in responding to the diseases in chickens due
to immuno disorders, phytotherapy offered by the
Ayurveda, and other traditional systems of medicine
appears to be potential part of the solution. The results
conclude that Ocimum sanctum and Argemone
mexicana modulate immune response and show
promising therapeutic value.
3 Material and Methods
3.1 Preparation of Hot Aqueous Extract (HAE)
The HAE of OS/AM was prepared using Soxhlet
apparatus as per the protocol described by Goel et al
(2008). The suspension was filtered through muslin
cloth and then what man No. 1 filter paper and dried
in lyophilizer under vacuum. Percentage yield was
14%~16% (w/w) for OS and 16%~19% (w/w) for AM
in terms of dried starting material.
3.2 Experimental birds
Standard pathogen free one day old chicks (Av. Wt
30~35 gm) were purchased from Uday hatchery,
Mathura and reared at poultry farm, DUVASU,
Mathura. All the birds were housed and fed under
standard conditions. Precautions were taken to prevent
the environmental contamination. In each experi-
mental group, individual bird identification was made
by using wing tag. Seven day old chicks were used for
the experiments. All these experiments were approved
by “Institutional Animal Ethics Committee” (IAEC),
and research was conducted under their guidelines. 6
birds per experimental group/control group were used.
3.3 Determination of nontoxic Dose (NTD) of hot
aqueous extract (HAE) of O. sanctum and A.
mexicana leaves
Hot aqueous extracts (HAE) of O. sanctum/A. mexicana
leaves were given orally to chickens. There were three
groups for OS as well as for AM for doses, 250 mg/kg,
500 mg/kg and 1000 mg/kg body weight, fed orally
for 21 days and one control group which was fed
placebo (Triple distilled water) in place of HAE for
determination of the safe and nontoxic dose. All the
birds were observed for apparent heath condition,
toxic signs, hematological changes and change in
body weight.
3.4 Humoral immune response
3.4.1 Preparation of S. enterica serovar Typhimurium
'O' antigen
S. enterica serovar Typhimurium “O” antigen was
prepared as described by Bhatia et al (2003). Smooth
colonies of S. enterica serovar Typhimurium grown on
brain heart infusion (BHI) agar medium were selected
and inoculated in BHI broth. Inoculated broth was
incubated for 6~8 hours at 37ºC and centrifuged at
3000 rpm for 20 minutes and supernatant was
discarded and washed with the normal saline and
boiled at 100ºC for 2.30 h. Heat inactivated S. enterica
serovar Typhimurium culture was employed as ‘O’
antigen for determination of humoral immune
response (HIR) in chickens.
3.4.2 Immunization of chickens
Three groups viz; control, O. sanctum and A.
mexicana HAE fed, each consisting 6 chickens of age
of 7 days were made. OS and AM groups of chickens
were fed orally with 250 mg/kg body weight of HAE
of OS/AM leaves for 21 days respectively. Control
group chickens were given orally triple distilled water
(placebo) for 21 days. On 22nd day 1st dose and seven
days apart 2nd dose of S. enterica serovar
Typhimurium ‘O’ antigen was given subcutaneously
to chickens of all the groups as shown in Table 3.
Serum samples were collected from each chicken of
all the groups before immunization, 7th day after first
immunization and 7 days after 2nd immunization for
determination of antibody titre against Salmonella ‘O’
antigen by indirect ELISA test.
3.4.3 Humoral immune response by measurement
of antibody titer by indirect ELISA:
The Salmonella “O” antigen coated wells of
polystyrene micro titer plate (Nunc) was incubated
overnight at 4. Following three washings with PBS
(PBS: pH 7.2, 0.01M) containing 0.05% tween-20
(PBS-T), blocking was done by 1% bovine serum
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Table 3 Immunization schedule
Days of immunization Control group O. sanctum fed group
A
. mexicana fed group
Dose (mL) Route Dose (mL) Route Dose (mL) Route
28th day of age (Ist dose) 0.5 s/c 0.5 s/c 0.5 s/c
35th day of age (IInd dose) 0.25 s/c 0.25 s/c 0.25 s/c
albumin dissolved in PBS, for 1½ hours at 37.
Serially diluted serum samples (1:100 to 1:10,240)
from 1 to 11 well of micro titer plate respectively 12th
well was used as negative control. Rabbit anti chicken
immunoglobulin–Y(IgY) conjugated with HRP (Horse
radish peroxidase, 1:4000) and substrate solution
TMB (1:20) were serially added to all the wells of
micro plate, the colour development was stopped after
15 minutes by adding of 50 µL of 1M sulphuric acid
to each well. The optical density was measured at
450~570 nm and compared with control group.
3.5 Cell mediated immune response in chickens
using DNCB
This study was designed and conducted to assess the
effect of HAE of OS and AM leaves on 2, 4-di nitro
chloro benzene (DNCB) sensitized chickens by the
method as described by Tiwari and Goel (1985). In
this experiment DNCB was used as an antigen. Three
groups having 6 chickens (7 day old) in each group
were made. Group II and III were fed orally with
250 mg/kg body weight of HAE of OS and AM leaves
extract respectively for 21 days. Group I served as
control group and placebo (TDW) was given orally for
21 days.
On 22nd day post feeding an aliquot of 0.25 mL of
DNCB (10 mg/mL) was applied drop by drop on left
side of un-feathered area of abdomen to individual
bird in each group and dried by blowing. The vehicle
(acetone) alone was applied on right side of abdomen.
After 7 days a challenge dose of (1 mg/mL) of DNCB
was applied on the same test side and vehicle alone on
right side of abdomen (Renu et al., 2003). The skin
thickness was measured by Varnier caliper before
challenge and 24 h, 48 h, 72 h post-challenge.
Difference in the thickness was calculated by
subtracting the thickness measured before challenge
from that of post challenge.
Authors’ contribution
All the authors contributed equally for this study. All the
authors read and approved the final version of the manuscript.
Acknowledgements
Authors are thankful to the University authorities for providing
necessary facilities to carry out this research work.
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... Goel et al, 2010 [87] hot aqueous extract of O. sanctum enhanced the antibody level by 42.85% in comparison to control group and acted as stimulant of humoral response stimulant; stimulatory effect on both arms of immune system Goel et al., 2008; [88] Varshney et al.,2013 [89] hot aqueous extract of Argemone mexicana enhanced the antibody level by 14.28% in comparison to control group and acted as stimulant of humoral response stimulant; antibody stimulation response but suppression in cell mediated immune response Goel et al., 2008; [88] Varshney et al.,2013 [89] rosemary powder and ethanolic extract failed to show any significant impact on antibody titers against NDV, SRBC and influenza disease virus, but remarkably improve total serum antioxidant activity. ...
... Goel et al, 2010 [87] hot aqueous extract of O. sanctum enhanced the antibody level by 42.85% in comparison to control group and acted as stimulant of humoral response stimulant; stimulatory effect on both arms of immune system Goel et al., 2008; [88] Varshney et al.,2013 [89] hot aqueous extract of Argemone mexicana enhanced the antibody level by 14.28% in comparison to control group and acted as stimulant of humoral response stimulant; antibody stimulation response but suppression in cell mediated immune response Goel et al., 2008; [88] Varshney et al.,2013 [89] rosemary powder and ethanolic extract failed to show any significant impact on antibody titers against NDV, SRBC and influenza disease virus, but remarkably improve total serum antioxidant activity. ...
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... The non-toxic dose was determined by oral feeding of three doses 250 mg/kg, 500 mg/kg and 1000 mg/kg body weight of HAE of OS and AM leaves. For in vivo antiviral study oral feeding of 250 mg/kg HAE of OS and AM leaves was done as it was ideal (Varshney et al., 2013). ...
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The present study was undertaken to explore the in vitro antiviral potential of hot aqueous extract (HAE) of Ocimum sanctum (OS) and Argemone mexicana (AM) leaves against Newcastle disease virus (NDV) and Infectious bursal disease virus(IBDV) in chicken embryo fibroblast (CEF) cell culture. First the nontoxic dose of HAE of OS and AM was determined in CEF. Doses of OS and AM below 20 mg/mL and 5 mg/mL respectively were found to be nontoxic to CEF in RPMI 1640 media. Anti NDV activity was determined by absence/lower cytopathic effect (CPE) in CEF and lower HA titer of cell culture supernatant. Anti IBDV activity was determined by absence/lower CPE in CEF. Besides in vivo antiviral effects of HAE of OS and AM against NDV and IBDV were evaluated in chicken model. 250 mg/kg body weight oral dose of HAE of OS and AM was found ideal and nontoxic in chickens and experimental chickens were fed this dose for 21 days for determination of in vivo antiviral effect. On 22 nd day respective groups of chickens were challenged orally with ID 50 dose of NDV and IBDV. OS and AM fed chickens challenged with either of the virus were better protected as compared to unfed controls.
... Argemone mexicana was reported to decrease the cell mediated immune response in albino rats (Goel et al., 2008). But increased humoral response against Salmonella antigen was found in chicken model (Varshney et al., 2013). ...
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Argemone mexicana, a prickly plant commonly called as prickly poppy is found in sub tropical regions and is well known for its medicinal properties. Its potential as a medicinal plant has been practiced traditionally and been prescribed as medicines by Ayurvedic, Unani, Siddha and Homeopathic practices since several years. Each part of plant posses bio active compounds that help in curing ailments like HIV, malaria, ring worm infections, fungal infections, cancer etc. These activities have been studied in vivo and in vitro set up and results have been obtained in favor. Further, phytochemical evaluation has unveiled the presence of compounds like berberine, argemonine, protopine etc, which show curative actions and could be used for treatment of diseases with future preception. This review is a sum up of all literature available through the internet and was searched using keywords ‘Argemone mexicana’, ‘phytochemical importance of Argemone’ and many other exclusive words with respect to different activities scanned for reviewing. The references provided in the papers were also given a thorough look and retrieved the respected data from them too. ‘Scopus’, ‘Pubmed’, ‘Google Scholar’, ‘Research Gate’ were used to search for the relevant papers through different journals available. The literature has then been framed in a way with up gradation about Argemone mexicana and its promising affects seen with the potential of the plant which is still undiscovered but could be utilized as curative methods
... Hashemipour, Kermanshahi, Golian, Veldkamp, & Hashemipour (2016) found an improved immune response in broilers fed a diet supplemented with thymol and carvacrol, characterized by an enhancement in hypersensitivity and an increase in total IgG and IgG anti-sheep red blood cells with a decreased heterophil to lymphocyte ratio. However, higher antibody titres in animals supplemented with plant extracts were also reported by other authors both in chickens (Franciosini et al., 2016;Hashemipour et al., 2013;Varshney, Dash, Goe, & Bhatia, 2013) and in ruminants (Ozkaya, Erbas, Ozkan, Baydar, & Aksu , 2017). ...
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The effects of an oregano and/or rosemary (Origanum vulgare L. and/or Rosmarinus officinalis L.) dietary supplementation to the diet of fattening pigs were investigated. Thirty-two grower-finisher pigs (45 kg) were divided into four dietary groups identified as: control diet (CTR); CTR+ 0.2% oregano (O); CTR + 0.2% rosemary(R), and CTR+ 0.1% oregano + 0.1% rosemary (OR). During the finishing period, all groups received a further supplementation of 0.5% of conjugated linoleic acids (CLA). Blood samples were collected after an adaptation period of 15 days to the new diet (T1) and at the end of the finishing period (T2) to evaluate antioxidant status (total antioxidant power and reactive oxygen metabolites) and immune responses (lymphocytic phenotyping and IgG levels). Pork meat samples were evaluated for glutathione peroxidase activity (GSHPx), total phenolic content and preference rating. A significant increase in B lymphocytes (CD79+) and a higher IgG level was observed in the R and O groups (P<0.05). Furthermore, there were significant effects of dietary supplementation on meat GSHPx activity and total phenolic contents (P<0.001 and P<0.005, respectively). Preference rating showed that pork derived from group R was the most preferred by the consumers.
... SX extract show the antibacterial [4], antifungal [5],Hypoglycemic [6], antifilariasis [7] and antioxidant [8] activity.Hematological parameters such as total WBC, RBC,hemoglobin and neutrophil are important constitute of immune system. An alteration in the concentration of these cells profoundly affects the health and immune system as they are known to recognize the foreign antigen and mount immune response [9].Present work was done to estabilish the correlation between the hematological and biochemical parameter with different pharmacological activity [10]. ...
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This work has been performed to study the efficacy of hot whole plant aqueous extract of Solanum xanthocarpum Schrad and Wendl on hematological and Biochemical parameters of wistar albino rats.Doses hot aqueous extract of different concentration like 125 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg body weight was given orally to different group of rats. As the result of dose administration level of hemoglobin , Packed cell volume (PCV), RBCs decreased and WBCs significantaly (p<.05) increased .On the other hand efficacy of dose on Biochmemical parameter show the mixed effect , the level of urea, creatinin ,albumin, and bilirubin as comparison with control wistar albino rat were not significantly differ while level of Glucose, total cholesterol, Alanin amono transferas (ALT) and Aspartate amino transferase (AST) significantly (p<.05 and .01) decreased .
... Hashemipour et al. (2013) found an improved immune response in broilers fed with a diet supplemented with thymol and carvacrol, characterized by an enhancement of hypersensitivity response and an increase of total IgG and IgG anti-sheep red blood cells with decreased heterophil to lymphocyte ratio. Varshney et al. (2013) examined the effects of Ocimum sanctum and Argemone Mexicana AEs in chickens, reporting higher antibody titres and improved cell-mediated immune response compared to control. With respect to the microbiological investigations, this study was mainly undertaken to detect possible differences in some bacterial populations following the administration of diets supplemented with AEs. ...
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A 57-day study was performed to determine the effects of two aqueous extracts (AEs) on broiler performance, immune function and intestinal microflora. Four groups of 75 one-day-old female broilers (Ross308) received one of the following treatments: (1) a standard commercial feed (C); (2) C supplemented with 2 g/kg rosemary AE (R); (3) C supplemented with 2 g/kg oregano AE (O); (4) C supplemented with 1 g/kg oregano AE + 1 g/kg rosemary AE (OR). Individual body weight, average daily gain, average daily feed intake and feed conversion efficiency were determined at 1, 11, 22, 36 and 57 days. Sample collections for IgG titration and intestinal microflora examination were performed at 22 and 57 days. The addition of oregano AE alone or in combination with rosemary AEs improved body weight up to 36 days of age (P < .01). A time effect was recorded for total serum IgG in all groups (P < .001) and the percentage increase of the value was positively (P < .05) influenced by the AE supplementation. Lactobacilli raised (P < .001) in ileum and cecum of all groups supplemented with AEs. Staphylococcus spp. population was constantly lower in both intestinal tracts of the AE supplemented groups. On the basis of our results, AEs could improve broiler performance and immune function and contribute to a balanced gut microflora, essential for the digestion process and protection against enteropathogenic organisms.
Thesis
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This report was written to advance understanding of the uses and benefits of the herb holy basil (Ocimum sanctum). This version was written in 2004 and updated information is found in the book: ADAPTOGENS.
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Immunomodulation as a therapy for revitalaizing the suppressed immune system is under extensive trials in human medicine to treat immunologically related disorders like arthritis, asthma and allergy. Large number of medicinal plants are being tested for the immunomodulatory effects, among which Ocimum sanctum (Tulsi) is in the forefront (Patwardhan et al., 1990). Infectious Bursal Disease (IBD) in poultry is also characterized by imunosuppression and subsequent secondary infections (Ajinkya et al., 1980). The present investigation reports the usefulness of O.sanctum dry leaves as immunomodulator in poultry, naturally infected with IBD virus.
Article
Effect of leaf extract of Ocimum sanctum on (i) the specific and non-specific immune responses and (ii) disease resistance against Aeromonas hydrophila was investigated in Oreochromis mossambicus. Sheep red blood cells (SRBC) and Heat aggregated bovine serum albumin (HA-BSA) were used as antigens for specific and non-specific immune response studies, respectively. Antigens were administered through intraperitoneal route. Anti-SRBC antibody titres were determined by direct haemagglutination and anti-bacterial antibodies were determined by bacterial agglutination. Nitroblue tetrazolium (NBT) assay was used to determine neutrophil activity. Disease resistance was measured in terms of relative percent survival (RPS). The immunostimulatory effect of the leaf extract of O. sanctum, when administered through intraperitoneal and oral routes was obvious. Leaf extract of O. sanctum, when administered intraperitoneally, stimulated both antibody response and neutrophil activity. Dietary intake of O. sanctum also enhanced the antibody response and disease resistance against A. hydrophila. Possibility of using O. sanctum as immunostimulant in the maintenance of finfish health in intensive freshwater aquaculture is suggested.
Article
A methanol extract and an aqueous suspension of Ocimum sanctum inhibited acute as well as chronic inflammation in rats as tested by carrageenan-induced pedal edema and croton oil-induced granuloma and exudate, respectively. In both test procedures, the anti-inflammatory response of 500 mg/kg of methanol extract and aqueous suspension was comparable to the response observed with 300 mg/kg of sodium salicylate. Both the extract and suspension showed analgesic activity in the mouse hotplate procedure and the methanol extract caused an increase in the tail-withdrawal reaction time of a subanalgesic dose of morphine. Both preparations reduced typhoid-paratyphoid A/B vaccine-induced pyrexia. The antipyretic action of the methanol extract and aqueous suspension was weaker and of shorter duration than that of 300 mg/kg sodium salicylate. Oral premedication with the methanol extract and the aqueous suspension delayed castor oil-induced diarrhoea in rats.
Article
Optimum conditions for the contact sensitivity (CS) test using 2,4-dinitrochlorobenzene (DNCB) were sensitization with 0.25 ml of a DNCB solution (10 mg/ml) and challenge with one-tenth of this dose two weeks later. This produced reaction in terms of skin thickness which could be easily observed and measured. The cutaneous hypersensitivity developed slowly, reaching its maximum manifestation after 24 h of challenge and gradually declining thereafter. The development of a skin reaction was typical of delayed type hypersensitivity and was characterized histopathologically by congestion, oedema, mononuclear and heterophilic cell infiltration in the dermal layer and lymphocytic perivascular cuffing. The efficiency of the CS test was compared with that of the lymphocyte transformation (LT) test and graft-versus-host (GvH) reaction for monitoring cell-mediated immunity (CMI) in normal and CMI deficient chickens. CMI deficient chickens were prepared by neonatal thymectomy and inoculation of antithymocyte serum. The CMI response of deficient chickens was significantly less (p less than 0.01) as determined by all three tests. The percentage decrease in response to mount GvH, CS and LT was 88.9, 58.3 and 74.6, respectively. All the tests were found to be equally effective in assessing CMI response in chickens as determined by comparing the means of their performances.
Article
The present study investigates the effect of Ocimum sanctum seed oil (OSSO) on some immunological parameters in both non-stressed and stressed animals. An attempt has also been made to explore the possible mechanism of immunomodulatory activity. OSSO (3 ml/kg, ip) produced a significant increase in anti-sheep red blood cells (SRBC) antibody titre and a decrease in percentage histamine release from peritoneal mast cells of sensitized rats (humoral immune responses), and decrease in footpad thickness and percentage leucocyte migration inhibition (LMI) (cell-mediated immune responses). Restraint stress (RS) produced a significant reduction in the anti-SRBC antibody titre, foot pad thickness and percentage LMI (% LMI). The effects of RS on humoral as well as cell-mediated immune responses were effectively attenuated by pretreating the animals with OSSO. Co-administration of diazepam (1 mg/kg, sc), a benzodiazepine (BZD), with OSSO (1 ml/kg, ip) enhanced the effect of OSSO on RS-induced changes in both humoral and cell-mediated immune responses. Further, flumazenil (5 mg/kg, ip), a central BZD receptor antagonist inhibited the immunomodulatory action of OSSO on RS-induced immune responsiveness. Thus, OSSO appears to modulate both humoral and cell-mediated immune responsiveness and these immunomodulatory effects may be mediated by GABAergic pathways.