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The effects of four different levels of wood beto-ny (WB) (Stachys lavandulifolia Vahl) extract (0, 2, 4 and 8% W/W in the diet) were investigated on some serum enzymatic activities and acute stress response of juvenile common carp (Cyprinus carpio). After 10 weeks post feeding, two groups of fish (0 and 8% WB) were subjected to handling and crowding stress and the stress responses including serum cortisol, glucose and some electrolytes (Na + , K + and Ca +2) levels were assayed. The results revealed that the fish fed on 8% WB had significantly lower levels of lactate dehydrogenase (LDH) and alanine transaminase (ALT) compared to the control group (P<0.05). Although cortisol and glucose levels were increased significantly shortly after stress (P<0.05), the elevation level was significantly lower for fish fed on 8% WB (P < 0.05). In control group, sodium level decreased (P < 0.05) at 3 h post stress but did not show any change in 8% WB (P > 0.05). The potassium and calcium levels showed insignificant fluctuations post stress (P>0.05). Dietary inclusion of WB seems to improve liver function and response to acute stress in juvenile common carp.
Iranian Journal of Aquatic Animal Health 2014
1 (1) 17-26
Effect of wood betony (Stachys lavandulifolia Vahl) extract on some
serum biochemical changes and acute stress response in juvenile
common carp (Cyprinus carpio)
S Bahrami Babaheydari, S Dorafshan, F Paykan Heyrati and N Mahboobi Sooani
Department of Natural Resources, Isfahan University of Technology, Isfahan, Iran
Received: March 2014 Accepted: July 2014
The effects of four different levels of wood beto-
ny (WB) (Stachys lavandulifolia Vahl) extract (0,
2, 4 and 8% W/W in the diet) were investigated on
some serum enzymatic activities and acute stress re-
sponse of juvenile common carp (Cyprinus carpio).
After 10 weeks post feeding, two groups of sh (0
and 8% WB) were subjected to handling and crowd-
ing stress and the stress responses including serum
cortisol, glucose and some electrolytes (Na+, K+
and Ca+2) levels were assayed. The results revealed
that the sh fed on 8% WB had signicantly lower
levels of lactate dehydrogenase (LDH) and alanine
transaminase (ALT) compared to the control group
(P<0.05). Although cortisol and glucose levels were
increased signicantly shortly after stress (P<0.05),
the elevation level was signicantly lower for sh
fed on 8% WB (P < 0.05). In control group, sodium
level decreased (P < 0.05) at 3 h post stress but did
not show any change in 8% WB (P > 0.05). The
potassium and calcium levels showed insignicant
uctuations post stress (P>0.05). Dietary inclusion
of WB seems to improve liver function and response
to acute stress in juvenile common carp.
Keywords: stress, enzyme activity, ion regulation,
wood betony, common carp.
Fish cultivation under intensive conditions has been
increased all around the world. High rearing den-
sities, poor water quality, weighing, transportation
and grading are common stressors which can both
affect sh welfare and increase the risk of disease
under intensive sh cultivation. Different levels of
stress reactions has been well-dened in sh includ-
ing primary (hormonal response), secondary (meta-
bolic and haematological changes) and tertiary as a
last stage which leads to exhaustion, growth decline
or death (Barton 2002). Urbinati & Carnerio (2001)
dened handling as a practice including chasing,
capture and exposing the sh to air which can cause
explicit physiological and endocrinological respons-
es in sh. Dobšikova, Svobodova, Blahova, Modra
& Velišek (2006) observed that 12 h transportation
could cause signicant changes in some biochemical
and haematological indices of three years old com-
mon carp. However, McCormick, Shrimpton, Car-
ey, O’Dea, Sloan, Moriyama & Björnsson (1988)
found out that handling stress decreased growth rate
of Atlantic salmon (Salmo salar). Different practic-
es such as water quality optimization, optimal sh
density and administration of natural or synthetic
compounds could be used to improve sh resistance
to stressor. Synthetic feed additives have some ad-
verse effects such as bioaccumulation and environ-
mental pollution. Due to these problems, much more
attention has been focused on the use of medicinal
plants in aquaculture (Citarasu 2010; Chakraborty
& Hancz 2011). Most of the studies about the effect
of using herbal biomedicine in aquaculture has been
focused on their application such as growth pro-
moter, immune stimulator or antipathogenic agent
in different cultivated sh species like African cat-
sh (Clarias gariepinus) (Dada & Ikuerowo 2009;
Soosean, Marimuthu & Sudhakaran 2010), Tilapia
Correspondence S Dorafshan, Department of Natural
Resources, Isfahan University of Technology, Isfahan, Iran
(e-mail: sdorafshan
(Orechromis mossambicus) (Immanuel, Uma &
Iyapparaj 2009), common carp (Cyprinus carpio)
(Alishahi, Ranjbar & Ghorbanpour 2010; Pakravan,
Hajimoradloo & Ghorbani 2012) and olive ounder
(Paralichthys olivaceus) (Cho & Lee 2012). How-
ever, little information is available on the effects of
herbs on stress responses in sh (Xie, Liu, Zhou,
Su, He, Pan, Ge & Xu 2008; Shahsavani, Baghshani
& Alishahi 2010). Wood betony (Stachys lavandu-
lifolia Vahl) which has been belonged to family
Lamiaceae, is grown in many parts of Iran, Turkey,
Iraq, Syria, Armenia as well as Georgia (Javidnia,
Mojab & Mojahedic 2004). Fresh and dried areal
parts such as leaves, owers and roots have been
used as traditional drugs for treatment of wounds
and bruises, mouth ulcers, gum inammations (Ody
1997) as well as treating arthritis and respiratory
inammatory disorders (Rezazadeh, Zaringhalam,
Manaheji & Kebryaeezadeh 2009). Alkanoids (in-
cluding stachydrine and trigonelline), tannins, sa-
ponines, nicotinic acid, polyphenols, organosulds
and steroids are the main components of wood bet-
ony (Vundac, Brantner & Plazibat 2007). Some of
the Stachys components’ have shown a variety of
biological activities (Chakraborty & Hancz 2011;
Ghasemi Pirbalouti, Jahanbazi, Enteshari, Malek-
poor & Hamedi 2010). The biological activity of
WB has not yet been studied in sh. Moreover, the
ability of herbal biomedicine on osmoregulation
and stress response has not been studied seriously.
Hence, this study was aimed to evaluate the effects
of dietary inclusion of WB extract on some serum
biochemical characteristics and stress responses in
juvenile common carp, which has been exposed to
handling and crowding stress.
Materials and Methods
Two hundred and fty juvenile common carp (35
± 5.2 g) were obtained from Isfahan Fish Propaga-
tion and Breeding Center in summer 2012. The sh
were kept under environmental condition, placed in
10 m3 rectangular concrete tanks for 2 weeks for ac-
climatisation. Then, they fed on a commercial carp
feed (Isfahan Mokkamel, Iran) with proximate com-
position (wet basis %) as the following conditions:
humidity (9.2%), protein (32%), lipid (10.2%) and
ash (11.1%). These amounts were obtained based
on our analyses (data not shown).
Plant extract
In spring 2012, the Wood betony aerial parts includ-
ing owers and leaves were collected from natural
habitat, Isfahan province. The plants were delivered
to the Central Herbarium of Isfahan University of
Technology, Department of Natural Resources for
nal identication and analyses. Hydro-alcoholic
plant extraction was done based on Ghasemi Pir-
balouti et al. (2010) with some modication. Brief-
ly, aerial parts of the plants were washed thoroughly
with distilled water and dried at room temperature
under shading. Then, the plants were grounded
into powder. A 100 g of powdered plant material
was soaked in 500 mL of ethanol (75%) for 48 h,
shacked vigorously to allow for proper extraction.
After ltering of the extract through Whatman pa-
per no. 1, ltrate was concentrated using a rotary
evaporator at 50 ˚C. Finally, 20 mL of concentrated
liquid extract was obtained from l00 g of the plant
powder; each mL of the concentrated extract was
almost equal to 5 g of the plant powder.
Feed preparation and feeding trails
In order to prepare the diets, the commercial pel-
let diet was crushed and mixed with the appropri-
ate WB liquid extract concentration (the extract
volumes were adjusted by adding distilled water to
nal volume of 100 mL for each kg of diet), remade
into the pellets, were allowed to be dried for 72 h
at room temperature and then stored at refrigerator
until it was used. After 2 weeks of acclimatization,
180 individual sh (44±0.62 g) were randomly di-
vided into four groups, with three replicate each.
The rst treatment was fed on normal diet without
S. lavandulifolia and kept as control group. The sec-
ond, third, and fourth ones were fed on normal diet
containing 2, 4 and 8 % of S. lavandulifolia, respec-
tively (dened as 2 WB, 4 WB and 8 WB). Each
replicate contained 15 individuals in a bereglass
tank (110 L water volume, 50% renewed each day).
S Bahrami Babaheydari et al., Effect of wood betony extract on stress response in carp
Iranian Journal of Aquatic Animal Health
Water quality was monitored throughout the exper-
imental period at daily intervals; temperature 25 ±
1°C, pH 7.21 ± 0.5 and dissolved oxygen concentra-
tion at 7.5 ± 0.06 mg L-1. Fish were fed frequently
on a diet of 32% crude protein (CP) at a rate of 2%
body weight, three times a day for 70 days.
Stress challenge
The stress challenge was performed according to
Tahmasebi-Kohyani, Keyvanshokooh, Nematolla-
hi, Mahmoudi & Pasha-Zanoosi (2011) with some
modications. Briey, at the end of 10 weeks, 30
sh from control and 8 WB were subjected to stress
challenge. For this purpose, the sh were remained
on experimental diets and acclimatized for 1 week
as dened before. After the acclimation period,
three sh per tank were removed for sampling (time
0) prior to subjecting the remaining sh in the tank
to an acute stress. The stress consisted of netting the
remaining sh from the tank, holding them out of
water for 5 min, and then crowded at an approxi-
mate density of 150 g L-1 in a plastic mesh bucket
in their original tank for 3 h without any aeration or
water exchange. The experimental sh were sam-
pled after 1 h of crowding, as they were released
from the crowding stress (3 h) and at 8, 24, and 48 h.
Blood sampling and biochemical analysis
At the end of the 10-week experiment, for biochem-
ical analyses, at least 3 sh from each replicate were
anaesthetized with clove powder (100 ppm) and
blood sampling (1.5-3 mL) was performed individ-
ually from caudal puncture. The blood was centri-
fuged at 3000 rpm for 10 min and the supernatant
serum was collected. The serum was kept frozen at
-80 °C until analyses for enzymatic activity of lac-
tate dehydrogenase (LDH), aspartate transaminase
(AST), alkaline phosphatise (ALP), and alanine
transaminase (ALT) activities (Peyghan & Azary
Takamy 2002).
To evaluate the stress response of the sh, they were
captured with minimal disturbance at the time de-
scribed above (stress challenge). Cortisol and glu-
cose levels were determined using radioimmunoas-
say method (Immunotech, France) and colorimetric
glucose oxidize procedure (Benfey & Biron 2000).
The ions levels were assayed based on Braun, Lima,
Baldisserotto, Dafre & Nuner (2010).
Statistical analysis
Statistical analysis was performed by one way ANO-
VA at 5% signicant level. A multiple comparison
test (Duncan multiple range test, DMRT) was con-
ducted to compare the statistically signicant dif-
ferences among the groups using SPSS Version19.
Values were presented as mean ± standard errors.
To evaluate the differences between stress related
analysed parameters at the same time between two
experimented groups, paried T-test was utilized.
At the end of the experiment (10 weeks), levels of
serum ALP and AST did not change signicantly
among groups (P>0.05). LDH showed signicant
decrease in group of sh fed on 8% WB (256.25
±40.30 UL-1) compared to control group (605.5±
78.98 UL-1) plus other groups treatments (Table 1;
The levels of ALT were in the range of 3.5-8.25
UL -1 (Table 1). Inclusion of WB in the diet, high-
er than 2% could decline ALT level signicantly
(Table 1; P<0.05). Regarding the stress challenges,
the sh treated by control diet had an initial cortisol
level of 9.23 ng mL-1 which rose signicantly to 27
ng mL-1 at 1 h and reduced to 16 ng mL-1 at 3 h and
remained in a plateau state without any signicant
changes until 48 h (Fig. 1).
The sh fed on 8% WB had an initial cortisol level at
6.53 ng mL-1 which increased signicantly to 18.66
ng mL-1 after 1h and then reduced to 12.2 ng mL-1
which were similar to basal level after 8 h (Fig. 1).
Cortisol level showed very limited changes during
8-48 h post stress in 8WB group (Fig. 1). In general,
the hormone levels were higher in sh fed control
diet than that of 8 % WB (Fig. 1). However, only
signicant differences were observed 1 h and 8 h
post-stress (Fig. 1; P<0.05). Glucose concentrations
in sh fed control diet were 62.66 mg dL-1 which
signicantly rose up to 157 and 264 mg dL-1at 1 h
and 3 h, respectively (Fig. 2; P<0.05).
As Figure 1 illustrates, the glucose level did not re-
turn to the basal level until 48 h post-stress (P<0.05).
In sh treated with 8% WB, the initial glucose level
was reported as 59.33 mg dL-1, that is very similar
to those which have been measured in control group
(Fig. 1; P > 0.05). The glucose concentration was el-
evated in a very gradual route and reached to 96 and
146.66 mg dL-1 at 1 and 3 h post-stress, respectively
(Fig. 1). Similar to those reported for control group,
the glucose level for 8WB sh did not changed sig-
nicantly during 3h to 48h (Fig. 1; P > 0.05). At all
intervals, the glucose levels were higher in control
group compared to 8 WB, but signicant differenc-
es were only observed at 1, 3 and 8 h post-stress
(Fig. 2; P < 0.05). At both treatments prior to stress
induction, serum sodium concentrations were 156
mmol L-1 in sh. In control group, the sodium level
dropped to the lowest level at 3 h, then elevated to
initial level throughout the experiment (Fig. 3A).
In sh fed on 8% WB, the sodium level did not
change signicantly during the experiment (time 0
to 48h; Fig. 3A; P > 0.05). In all sampling times, the
sodium levels in sh fed on 8% WB were higher
than those of measured in control group (Fig. 3A),
but these differences were insignicant (Fig. 3A;
P>0.05). For control group, the potassium and cal-
cium levels were in the range of 0.53-0.73 and 2.55-
3 mmol/L, respectively and did not show any sig-
nicant differences during the sampling time (Fig.
3B, C; P > 0.05). Similar to the control group, the
concentration of potassium and calcium levels were
too low, that is, in the range of 0.6-1 and 2.81-2.94
mmol/L, respectively. The concentrations of these
ions did not change throughout the experiment (Fig.
3B, C; P > 0.05). At the most sampling time, the
potassium and calcium levels for sh fed on 8WB
were higher than those measured for control sh
(Fig. 3B, C) without any signicant differences
Variable (U/L-1)* Control (0) 2% 4% 8%
ALP 87.75 ± 6.68a81.75 ± 11.23a80.50 ± 6.68a78.50 ± 1.19a
AST 178.00 ± 51.96a115.00 ± 35.12a114.50 ± 17.93a70.75 ± 11.98a
LDH 605.50 ± 78.98a580.25 ± 106.08a557.00 ± 102.10a256.25 ± 40.30b
ALT 8.25 ± 1.60a6.75 ± 0.75ab 4.75 ± 0.85bc 3.50 ± 0.28c
Table 1 Serum enzymes (UL) in common carp fed different levels of dietary wood betony extract for 10 weeks
Alkaline phosphatase, ALP; asparate transaminase, ASP; lactate dehydrogenase, LDH and alanine transaminase, ALT. Values are mean
± SEM. Mean values with different superscripts are signicantly different from each other (P<0.05).
Figure 1 Serum cortisol (ng mL-1) levels of common carp sub-
jected to acute handling followed by 3 h of crowding stress in
sh fed the control and 8% diets. Data are presented as mean
± standard error as error bars; Signicant differences between
different time of blood sampling in the same group (P<0.05;
Duncan test) or between two groups (0 and 8% of WB) at the
same time (P<0.05; t-test) are indicated by unlike letters, and
Figure 2 Serum glucose (mg dL-1) levels of Common carp sub-
jected to acute handling followed by 3 h of crowding stress in
sh fed the control and 8% diets. Data are presented as mean
± standard error as error bars; Signicant differences between
different time of blood sampling in the same group (P < 0.05;
Duncan test) or between two groups (0 and 8% of WB) at the
same time (P < 0.05; t-test) are indicated by unlike letters and
*, respectively.
S Bahrami Babaheydari et al., Effect of wood betony extract on stress response in carp
Iranian Journal of Aquatic Animal Health
(Fig. 3B, C; P > 0.05).
Many research works have been conducted on the
use of different herbal medicines in aquaculture both
in nsh (Chakraborty & Hancz 2011) and shellsh
species (Citarasu 2010). Despite many studies have
been investigated the effects of dietary phytochem-
ical on growth or immunity parameters, less infor-
mation is available about the use of these environ-
mental friendly compounds such as anti-stress agent
in aquaculture industry. Different stressors can in-
uence hypothalamus- pituitary- internal axis of the
aquatics, mainly sh like other vertebrates and im-
pact on dynamic balance of organisms. In this study,
at least the level of two serum enzymes including
LDH and ALT were signicantly reduced by inclu-
sion of 8 % WB in the diet of carp. Also sh fed
with 2% and 4% WB had a tendency of decrease in
these enzymes. None signicant decreasing pattern
was also reported for ALP and AST by elevating the
WB in the diet. Although mentioned enzymes have
been obtained in different tissues such as heart, skel-
etal muscle and kidneys in vertebrates, liver usual-
ly considered as a main source of these enzymes.
Also, the elevation of these enzymes in sh serum,
in response to exogenous compounds such as food
additive and heavy metals, may be considered as
an indication for liver dysfunction (Talas & Gulhan
2009). The decline of some serum enzyme at least
in groups received the highest dose of WB could
explain potential benet of wood betony for sh
welfare by improving liver function. Recently, Yıl-
maz, Ergün & Çelik (2013) revealed the positive ef-
fects of dietary thyme (Thymus vulgaris), rosemary
(Rosmarinus ofcinalis) and fenugreek (Trigonella
foenum graecum) on some physiological character-
istics of sea bass (Dicentrarchus labrax). The sh
fed on these medicinal herbal extracts had better liv-
er function, which has been evaluated as lower total
liver fat, visceral fat index and hepatosomatic in-
dex (HSI). Some researchers (e.g., Cho, Lee, Park,
Ji, Lee, Bae & Oh 2007; Zheng, Tan, Liu, Zhou,
Xiang & Wang 2009) conducted experiments with
channel catsh, Ictalurus punctatus, fed a basal diet
containing Greek oregano, Origanum heracleoti-
cum essential oil and olive ounder, Paralichthys
olivaceus, fed diets supplemented with green tea,
Camellia sinensis, respectively and showed signi-
Figure 3 The effects of dietary wood betony extract on some
ion concentrations Na+ (A), K+ (B) and Ca++ (C) in common
carp exposed to acute handling followed by 3 h of crowding
stress. Data are presented as mean ± standard error as error
bars; Signicant differences between different time of blood
sampling in the same group (P<0.05; Duncan test) is indicated
by unlike letters.
cant improvement in the sh liver function. Further-
more, Srinivasan (2005) reported different active
compounds of herbs stimulated digestion, accom-
panied by enhanced bile acid concentration, stim-
ulated the pancreas, and increased the secretion of
digestive enzyme activities in chicken broilers. It is
possible that different antioxidant compounds avail-
able in the wood betony extract can inuence liver
function in sh. Frankiˇc, Voljˇc, Salobir & Rezar
(2009) reported that antioxidants enhance the syn-
thesis of bile acids in the liver; However, due to the
limited scientic data about the effect of herbal bio-
medicine on liver enzyme activity (Xie et al. 2008),
more research is needed to expand our knowledge
concerning the physiological mechanisms of sh
It is well-documented that sh like other vertebrate
try to cope stress by changing some biochemi-
cal compositions such as blood cortisol, glucose
as well as different ions (Fevolden & Roed 1993;
Barton 2002). Cortisol elevation usually considered
as a rst smart signal for sh under stress (Hsieh,
Chen & Kuo 2003). The blood cortisol level of
common carp was as low as 10-20 ng mL-1 before
stress challenge, abruptly increased just 1 h after
stress exposure, and the sharply decreased to bas-
al level at 3 or 8 h. Regardless, the pattern of the
hormone changes, the blood cortisol levels of sh
fed on 8% WB apparently reported lower than the
control group in the most time intervals. This indi-
cates that WB can mitigate cortisol charge, which
is motivated by handing stress. These ndings are
in conformity with the report of Xie et al. (2008)
on common carp. Blood glucose elevations imply
to stress usually considered as atypical secondary
stress response (Barton & Iwama 1991). Usually,
the major reason for blood glucose rise is dissimila-
tion and dissolution of liver glycogen for providing
energy during stress process, caused by elevation
of different hormone level, especially in cortisol
(Hsieh et al. 2003). The current study indicated that
blood glucose levels increased signicantly after
stress and reached the highest concentration at 3 h;
2 h after highest level of blood cortisol level. None-
theless, the augmentation of blood sugar in sh fed
on 8% WB was much lower than control group in-
dicating that WB could also assuage blood glucose
load probably because of the hormone diminish.
Signicant reduction in serum glucose levels were
also reported for Tilapia (Oreochromis mossambi-
cus) fed on diet supplemented from four different
medicinal plant (Immanuel et al. 2009) and com-
mon carp received diet containing rhubarb (Rheum
ofcinale) extract (Xie et al. 2008). Ion concentra-
tions could be used deliberately as a useful tool to
evaluate secondary stress response in sh. It is un-
der control of several neuroendocrine pathways in
sh which some of them are similar to those affect
stress responses. It is well understood that releasing
cathecholamines and cortisol can modify ion reg-
ulations (McCormick 2001). In this study, the ion
concentration level leaved unaffected in both con-
trol and 8WB all tested interval. However, the only
exception was lower sodium concentration of the
control sh at 3 h in comparison to time zero which
may show some positive effects of dietary WB on
sodium regulation in common carp. Nevertheless,
the ion disturbance in sh, after stress exposure,
has been inuenced by several factors such as type
of stress, stress intensity, duration, and sh species
(Barton 2002; Tejpal, Pal, Sahu, Kumar, Muthappa,
Vidya & Rajan 2009). Based on our knowledge, the
effects of dietary phytochemical on ion regulatory
capacity of sh are poorly understood (Immanuel et
al. 2009) and further research is needed in order to
nd out the exact mechanisms.
In conclusion, the ndings of this study indicated
that a dietary addition of wood betony could im-
prove some of the physiological conditions of com-
mon carp. Thus, this study provides a new perspec-
tive for the use of medicinal herbs, which can be
added to sh feed literature. Wood betony contain-
ing a number of different active components, which
possibly play an important role in improving organ
function and stress responses. However, the mech-
anism by which this occurs needs further and more
detailed research.
The authors would like to thank to Isfahan Aquatic
S Bahrami Babaheydari et al., Effect of wood betony extract on stress response in carp
Iranian Journal of Aquatic Animal Health
Propagation and Breeding Center for supplying sh.
This research was supported with funding from the
Isfahan University of Technology under grant num-
ber 502.90.53949 awarded by Dr. Salar Dorafshan.
We also thank Dr. Yazdan Keivany for reviewing
the manuscript.
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ﻰﯾﺎﯿﻤﯿﺷﻮﯿﺑ ىﺎﻫﺮﺘﻣارﺎ ﻰﺧﺮﺑ ﺮﺑ (Stachys lavandulifolia) ﻰﻫﻮﮐ ىﺎﭼ هرﺎﺼﻋ ﺮﺛا
ناﻮﺟ(Cyprinus carpio) ﻰﻟﻮﻤﻌﻣ رﻮﭙﮐ رد دﺎﺣ سﺮﺘﺳا ﻪﺑ ﺦﺳﺎﭘ و مﺮﺳ
3ﻰﻧﺎﯿﻓﻮﺻ ﻰﺑﻮﺒﺤﻣ اﺮﺼﻧ و 2ﯽﺗﺮﯿﺣ نﺎﮑﯿﭘ ﻪﻤﻃﺎﻓ ،*2
نﺎﺸﻓارد رﻻﺎﺳ ،1ىرﺪﯿﺣﺎﺑﺎﺑ ﻰﻣاﺮﻬﺑ ﺪﻤﺻ
ناﺮﯾا ،نﺎﻬﻔﺻا ،نﺎﻬﻔﺻا ﯽﺘﻌﻨﺻ هﺎﮕﺸﻧاد ،ﯽﻌﯿﺒﻃ ﻊﺑﺎﻨﻣ هﺪﮑﺸﻧاد ،تﻼﯿﺷ هوﺮﮔ ،ﺪﺷرا ﯽﺳﺎﻨﺷرﺎﮐ ﻪﺘﺧﻮﻣآ ﺶﻧاد 1
ناﺮﯾا ،نﺎﻬﻔﺻا ،نﺎﻬﻔﺻا ﯽﺘﻌﻨﺻ هﺎﮕﺸﻧاد ،ﯽﻌﯿﺒﻃ ﻊﺑﺎﻨﻣ هﺪﮑﺸﻧاد ،تﻼﯿﺷ هوﺮﮔ ،رﺎﯾدﺎﺘﺳا 2
.ناﺮﯾا ،نﺎﻬﻔﺻا ،نﺎﻬﻔﺻا ﯽﺘﻌﻨﺻ هﺎﮕﺸﻧاد ،ﯽﻌﯿﺒﻃ ﻊﺑﺎﻨﻣ هﺪﮑﺸﻧاد ،تﻼﯿﺷ هوﺮﮔ دﺎﺘﺳا 3
و ﻰﻣﺮﺳ ىﺎﻫ ﻢﯾﺰﻧآ ﻰﺧﺮﺑ ﺢﻄﺳ ﺮﺑ هﺮﯿﺟ رد ﺪﺻرد 8 و 4 ،2 ،0 ﻞﻣﺎﺷ (Stachys lavandulifolia) ﻰﻫﻮﮐ ىﺎﭼ هرﺎﺼﻋ ﻒﻠﺘﺨﻣ ﺢﻄﺳ رﺎﻬﭼ ﺮﯿﺛﺎﺗ
8 و 0) هوﺮﮔ 2 ،ﻪﺘﻔﻫ 10 تﺪﻣ ﻪﺑ ﻰﻫداﺬﻏ هرود نﺎﯾﺎﭘ رد .ﺖﻓﺮﮔ راﺮﻗ ﻰﺳرﺮ درﻮﻣ (Cyprinus carpio) ناﻮﺟ ﻰﻟﻮﻤﻌﻣ رﻮﭙﮐ رد دﺎﺣ سﺮﺘــﺳا ﻪﺑ ﺦــﺳﺎﭘ
ﻰﺧﺮﺑ حﻮﻄﺳ و ﺰﮐﻮﻠﮔ ،لوﺰﯿﺗرﻮﮐ ﺮﯾدﺎﻘﻣ ﻞﻣﺎﺷ مﺮﺳ ﻰﺳﺮﺘﺳا ىﺎﻫ ﺦﺳﺎﭘ و ﺪﻨﺘﻓﺮﮔ راﺮﻗ ﻢﮐاﺮﺗ و ىرﺎﮑﺘــﺳد سﺮﺘــﺳا ضﺮﻌﻣ رد (ﻰﻫﻮﮐ ىﺎﭼ هرﺎﺼﻋ ﺪﺻرد
تﺎﺘﮐﻻ زا ىﺮﺘﻤﮐ ﺮﯾدﺎﻘﻣ ىراد ﻰﻨﻌﻣ رﻮﻃ ﻪﺑ ﺪﻧدﻮﺑ هﺪﺷ ﻪﯾﺬﻐﺗ ﻰﻫﻮﮐ ىﺎﭼ هرﺎﺼﻋ ﺪﺻرد 8 ﺎﺑ ﻪﮐ ﻰﻧﺎﯿﻫﺎﻣ .ﺪﺷ ﻰﺑﺎﯾزرا (Ca2+ و Na+ ، K+) ﺎﻫ ﺖﯿﻟوﺮﺘﮑﻟا
ﺰﮐﻮﻠﮔ و لوﺰﯿﺗرﻮ ناﺰﯿﻣ ،ﻰﺸﯾﺎﻣزآ هوﺮﮔ ود ﺮﻫ رد .(P<0/05) ﻧداد نﺎﺸﻧ ﺪﻫﺎﺷ هوﺮﮔ ﺎﺑ ﻪﺴﯾﺎﻘﻣ رد ار (ALT) زﺎﻨﯿﻤﺴﻧاﺮﺗ ﻦﯿﻧﻻآ و (LDH) زﺎﻧژورﺪﯿﻫد
زا ىﺮﺘﻤﮐ ﺮﯾدﺎﻘﻣ ،ﻰﻫﻮﮐ ىﺎﭼ هرﺎﺼﻋ ﺪﺻرد 8 ىوﺎﺣ هﺮﯿﺟ ﺎﺑ هﺪﺷ ﻪﯾﺬﻐﺗ نﺎﯿﻫﺎﻣ دﻮﺟو ﻦﯾا ﺎﺑ ،(P<0/05) ﺖﻓﺎﯾ ﺶﯾاﺰﻓا سﺮﺘﺳا زا ﺲﭘ ﻰﮐﺪﻧا تﺪﻣ مﺮﺳ
ﻰﻟﺎﺣ رد (P<0/05) ﺖﻓﺎﯾ ﺶﻫﺎﮐ سﺮﺘﺳا زا ﺲﭘ ﺖﻋﺎﺳ 3 ﻢﯾﺪﺳ ناﺰﯿﻣ ﺪﻫﺎﺷ هوﺮﮔ رد (P<0/05) . ﺪﻧداد نﺎﺸﻧ ﺪﻫﺎﺷ هوﺮﮔ ﺎﺑ ﻪﺴﯾﺎﻘﻣ رد ار ﺐﯿﮐﺮﺗ ود ﻦﯾا
و ﻢﯿﺳﺎﺘﭘ ،ﻰﺸﯾﺎﻣزآ ىﺎﻫ وﺮﮔ زا ﮏﯾ ﭻﯿﻫ رد .(P>0/05) ﺪﺸﻧ هﺪﻫﺎﺸ ﻰﻫﻮﮐ ىﺎﭼ هرﺎﺼﻋ ﺎﺑ هﺪﺷ ﻪﯾﺬﻐﺗ نﺎﯿﻫﺎﻣ مﺮﺳ ﻢﯾﺪﺳ ناﺰﯿﻣ رد ىراد ﻰﻨﻌﻣ ﺮﯿﯿﻐﺗ ﻪﮐ
دﺮﮑﻠﻤﻋ دﻮﺒﻬﺑ ﻪﺑ ﺮﺠﻨﻣ ىدوﺪﺣ ﺎﺗ ﺪﻧاﻮﺗ ﻰﻣ ﻰﻫﻮﮐ ىﺎﭼ هرﺎﺼﻋ ﻪﮐ ﺪﺳر ﻰﻣ ﺮﻈﻧ ﻪﺑ ﺞﯾﺎﺘﻧ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ .(P>0/05) دﺮﮑﻧ ﺮﯿﯿﻐﺗ سﺮﺘﺳا ﺮﯿﺛﺎﺗ ﺖﺤﺗ مﺮﺳ ﻢﯿﺴﻠﮐ
.دﻮﺷ ﻰﻟﻮﻤﻌﻣ رﻮﭙﮐ ناﻮﺟ نﺎﯿﻫﺎﻣ رد سﺮﺘﺳا ﻪﺑ ﺦﺳﺎﭘ و ﺪﺒﮐ
.ﻰﻟﻮﻤﻌﻣ رﻮﭙﮐ ،ﻰﻫﻮﮐ ىﺎﭼ ،ﻰﻧﻮﯾ ﻢﯿﻈﻨﺗ ،ﻰﻤﯾﺰﻧآ ﺖﯿﻟﺎﻌﻓ ،سﺮﺘﺳا :ىﺪﯿﻠﮐ ىﺎﻫ هژاو :لﻮﺌﺴﻣ ﻩﺪﻨﺴﯾﻮﻧ*
... Significant decrease in FCR and higher growth performance in Nile tilapia was reported when fed on diet supplemented with grape seed proanthocyanidins (Zhai et al., 2014). Babaheydari et al. (2014) observed that with increase in supplementation of Stachys lavandulifolia up to 40 mg/ kg, there was an increase in growth performance of tilapia. Better growth performance was reported in Cyprinus carpio when fed on diet supplemented with marshmallow extracts (Fallahpour et al., 2014). ...
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Authors' Contribution MA conducted the study and wrote the manuscript. SMH acquired funds, administered and supervised the project. MA and MH curated and edited the data. MZHA, ZY and AAB edited and reviewed the manuscript. A 70-days growth trial was carried out to ascertain how dietary supplementation of polyphenols with soybean meal-based diet influences the growth, antioxidant status and carcass composition of Labeo rohita fingerlings. Seven experimental diets were formulated with graded levels, 0, 50, 100, 150, 200, 250 and 300 mg/kg of polyphenols supplementation. Each test diet was allocated randomly to triplicate tanks with 15 fingerlings in every tank. These were fed @ 5% of live wet body weight. Results showed that L. rohita fingerlings fed on test diet T 4 with dietary supplementation of 150 mg/kg polyphenol showed significant (p<0.05) increase in growth performance having weight gain% (233.24%) and better feed conversion ratio (1.44). Similarly, significantly (p<0.05) improved body composition with crude protein (17.96%), crude fat (8.72%), ash (4.94%) and moisture (68.43%) were also recorded at same polyphenols supplementation level. However, in terms of antioxidant activity, increasing trend in inhibition of oxidation was recorded with increase in supplementation of polyphenols in test diets, having minimum oxidation (7.66) in fingerlings fed on T 7 with 300 mg/kg level of polyphenols supplementation.
... Therefore, its reduction by FFC can show relieved stress and maybe a booster for immune responses. Similarly, some researchers reported that the medicinal compounds like oxytetracycline hydrochloride, benzylpenicillin sodium, cefazolin, and neomycin sulfate [12], or the herbal alternatives such as Rhodomyrtus tomentosa [49], Stachys lavandulifolia [51], and Nectandra grandiflora [52] extracts show reducing effects on glucocorticoids in diseased rainbow trout, common carp and silver catfish, respectively. In our study, measurement of the enzymatic markers of liver and kidney function including ALT, ALP, and CK pointed out a change in challenged groups which was ameliorated by FFC. ...
... Therefore, its reduction by FFC can show relieved stress and maybe a booster for immune responses. Similarly, some researchers reported that the medicinal compounds like oxytetracycline hydrochloride, benzylpenicillin sodium, cefazolin, and neomycin sulfate [12], or the herbal alternatives such as Rhodomyrtus tomentosa [49], Stachys lavandulifolia [51], and Nectandra grandiflora [52] extracts show reducing effects on glucocorticoids in diseased rainbow trout, common carp and silver catfish, respectively. In our study, measurement of the enzymatic markers of liver and kidney function including ALT, ALP, and CK pointed out a change in challenged groups which was ameliorated by FFC. ...
... immunostimulating and antistress effects(Bahrami Babaheydari, Dorafshan, Paykan Heyrati, & Mahboobi Soofiani, 2015, Giannenas et al., 2012, Gobi et al., 2016, Mullally et al., 2017, Saccol et al., 2018, Tang et al., 2014, Xie et al., 2008.Serum aminotransferase enzymes are indicators of fish tissue health(Yousefi, Hoseinifar, Ghelichpour, & Hoseini, 2018). ...
The aim of the present study was to investigate the effects of thyme extract (TE) and oxytetracycline (OTC) on rainbow trout (Oncorhynchus mykiss) stress response, immune parameters, oxidative stress and enzymatic changes. The fish were assigned into eight treatments receiving diets with OTC (0 and 2.5 g/kg) and TE (0, 5, 10 and 20 g/kg) for 2 weeks. Thereafter, serum characteristics and gut oxidative status were evaluated. OTC significantly increased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and cortisol and glucose levels; however, TE significantly suppressed these changes. OTC significantly decreased, but TE significantly increased gut catalase (CAT) and glutathione peroxidase (GPx) activities. OTC significantly decreased gut superoxide dismutase (SOD) and increased gut glutathione‐s‐transferase (GST) activities, malondialdehyde (MDA) content and blood monocyte percentage; TE significantly mitigated these changes. TE significantly increased serum total protein, total Ig, lysozyme and ACH50, and blood WBC; however, OTC significantly decreased serum lysozyme and ACH50 activities and blood WBC and neutrophil percentage. In conclusion, OTC administration causes immunosuppression, which might be related to deterioration of fish health. TE augments trout innate immune and antioxidant status and is capable to counteract OTC‐induced health and immune deterioration.
... Antioxidants derived from plants, as polyphenols and flavonoids, may increase the effectiveness of vitamins [42] and may have compensatory effects when the vitamin levels are low [43]. In agreement, Antache et al. [44] stated that administration of thyme in combination with vitamin E had a synergistic effect on growth performance and reducing lipid peroxidation in Nile tilapia Mekkawy et al. [45] were also emphasized on the probable additive or synergistic effects of tomato paste carotenoids and vitamin E protecting from harmful effects of cadmium in Nile tilapia (Table 1) [46][47][48][49][50][51][52][53]. ...
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The ban on nutritive antibiotic use in Europe and the increased awareness of the consumers triggered a need for natural and safe feed additives to achieve better production results of farm animals. Plant extracts are used in animal nutrition as appetite and digestion stimulants, stimulants of physiological functions, for prevention and treatment of certain pathological conditions, as colorants and antioxidants. This article is a review of present literature data on the usage of plant extracts in poultry, pig and ruminant nutrition.
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Phytochemicals such as alkaloids, flavonoids, pigments, phenolics, terpenoids, steroids and essential oils are a large group of plant‐derived compounds commonly found in diets high in fruits, vegetables, beans and cereals. Plant remedies are closely linked to the maintenance of good health in the cultural heritage of many countries. Despite the potential benefits to health and performance as noted in various terrestrial animals, the use of phytochemicals in fish farming has rarely been investigated. Fish culture is under pressure to decrease the use of synthetic antibiotics and chemotherapeutics because of the risk caused to humans by chemical residues in food and by antibiotic resistance being passed on to human pathogens. Consequently, efforts are being made to exploit plants, plant extracts or natural plant compounds as potential alternatives to synthetic chemicals for the stimulation of immune responses and disease resistance in fish. The phytochemicals contained in herbs may enhance the innate immune system and possess antimicrobial capabilities that may be of immense use in fish culture without causing any environmental and/or hazardous problems. Most phytochemicals are redox active molecules that have anti‐oxidant characteristics that may improve the general physiological condition of fish. This review discusses the results emanating from different studies related to the in vivo application of plants and/or phytochemicals in fish in relation to immunostimulation, antipathogenic and antistress activities. Special emphasis is given to the use of Chinese and Indian herbs as immunostimulants in different finfish species during culture and as antibacterial agents against Aeromonas hydrophila.
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This study was conducted in order to investigate the effects of dietary thyme Thymus vulgaris , rosemary Rosmarinus officinalis , and fenugreek Trigonella foenum graecum as feed additives on total liver fat levels and biometric indices of Sea Bass Dicentrarchus labrax . Four isonitrogenous (48% crude protein) and isocaloric (21 kJ/g) diets were formulated to contain 0% (control), or 1% of thyme, rosemary, or fenugreek. In a 45‐d feeding trial, 12 fiberglass tanks (140 L) were each stocked with 17 fish (20.43 ± 0.03 g). Herbal supplemented diets significantly decreased the total liver fat, visceral fat index, viscerosomatic index, hepatosomatic index, and increased the spleen–somatic index and bile–somatic index ( P < 0.05). In conclusion, the results indicated that a dietary level of 1.0% for thyme, rosemary, or fenugreek could improve some of the physiological parameters of Sea Bass. Thus, this work provides a new perspective for the use of medicinal herbs that can be added to fish feed to improve the status of fish organs. Thyme, rosemary, and fenugreek contain a number of different active components, which possibly play an important role in improving these conditions, but the mechanism by which this is achieved needs further and more detailed study. Received July 19, 2012; accepted January 15, 2013
This study tested the onion powder (OP) supplementation in the diet of the olive flounder on the growth, body composition, and lysozyme activity. Thirty‐five fish averaging 5.1 g were randomly stocked into 18 individual 180‐L flow‐through tanks. A commercially available OP was used as a dietary additive. Six experimental diets were prepared to contain OP at the concentrations of 0, 0.5, 1, 2, 3, and 5% for diets OP‐0, OP‐0.5, OP‐1, OP‐2, OP‐3, and OP‐5, respectively. After the 8‐wk feeding trial, 20 fish from each tank were infected with Edwardsiella tarda and mortality was monitored for the following 96 h. No distinctive improvement in survival, weight gain, or feed efficiency of fish was observed at the end of the 8‐wk feeding trial. Lysozyme activity in fish fed the OP‐0.5 diet was higher than that of fish fed the OP‐0, OP‐1, OP‐2, OP‐3, and OP‐5 diets. The cumulative mortality of fish fed the OP‐0.5, OP‐1, OP‐2, OP‐3, and OP‐5 diets was lower than that of fish fed the OP‐0 diet at 72 h after E. tarda infection. Dietary inclusion of 0.5% OP was effective at improving lysozyme activity of fish, and OP seemed to be an effective immunostimulant to lower mortality upon E. tarda infection.
The present study was conducted to evaluate the use of Epilobium hirsutum extract in diet of common carp, Cyprinus carpio (20 ± 2 g). Different levels of plant extract (0%, 0.5%, 1, 3% and 3%+2‰ multivitamin (2 g of multivitamin per kg diet) were spread on commercial diet. The feed was offered by 8 weeks. Results showed that fish fed experimental diets had no significant difference (P>0.05) in specific growth rates, condition factor, feed conversion ratios and survival compared with control. There were no significant difference (P>0.05) in moisture, crude protein, crude lipid and ash content of common carp fed diets containing various levels of plant extract. The mortality of fish challenged by Aeromonas hydrophila have been recorded for 30 days after challenging, results showed that mortality decreased significantly (PPP>0.05) in infected and uninfected groups compared with the control.
Seven Croatian Stachys taxa (S. alpina, S. officinalis, S. palustris, S. recta subsp. recta, S. recta subsp. subcrenata, S. salviifolia and S. sylvatica) have been investigated in order to determine their content of biologically active compounds (polyphenols, tannins, phenolic acids and flavonoids) as well as their antioxidant activity. All taxa tested had a high content of total polyphenols, medium content of total phenolic acids, and a rather low content of tannins and flavonoids. The total phenolic acids content correlated significantly with total polyphenols content and the content of polyphenols unadsorbed on hide powder. A low correlation between plant phenolic acids and tannins was observed. Methanolic, ethanolic and dichloromethane extracts were investigated using DPPH, lipid peroxidase and xanthine oxidase assays. The extracts showed no inhibitory effects against lipid peroxidation and xanthine oxidase, but they had the ability to scavenge DPPH. The most effective in the DPPH assay were methanolic extracts of S. recta sub. recta and S. palustris whose radical scavenging activity was higher then that of the reference rutin. A low correlation between radical scavenging capacities of extracts with total flavonoids content was observed. The results indicate that investigated Stachys taxa exhibit potent antiradical activity and therefore could be a potential material for extracting free radical scavengers.
The safety of the consumption of spices – turmeric, red pepper and black pepper and their respective active principles was established in animal studies by observing the influence on growth, organ weights, nitrogen balance and blood constituents upon feeding at levels close to human intake as well as upto 100-times the normal human intake. Exhaustive animal studies documented the beneficial influence of turmeric/curcumin, red pepper/capsaicin, and garlic on lipid metabolism, especially anti-hypercholesterolemic effect of the three spices and anti-lithogenic effect of curcumin and capsaicin. The anti-diabetic effects of turmeric/curcumin, onion and cumin seeds were also evidenced with particular ameliorative influence of curcumin and onion on diabetic nephropathy. The antioxidant effects of curcumin (of turmeric), capsaicin (of red pepper) and eugenol (of clove) were evidenced both in in vitro and in vivo systems and the consequential health beneficial anti-inflammatory influence in experimentally induced arthritis was documented. The mechanism of digestive stimulant action of common spices examined in experimental animals revealed to be mediated through phenomenal stimulation of bile secretion with an enhanced bile acid concentration (ingredients essential for fat digestion and absorption) and an appropriate stimulation of the activities of digestive enzymes of pancreas and small intestine. The protective influence of hypolipidemic spices – curcumin, capsaicin and garlic on the altered fluidity of erythrocytes under hypercholesterolemic situation was evidenced in experimental animal models. Antioxidant spices were also shown to greatly reduce LDL-oxidation and also modulate the synthesis of prostaglandins and leukotrienes. Several spices or their extracts were also found to beneficially inhibit platelet aggregation. All these observations strongly indicate that many spices and their active principles are excellent nutraceuticals.
Stress in fish caused by physical disturbances encountered in aquaculture, such as handling and transport, evokes a variety of responses that may be adaptive or maladaptive. The overall effect of stress may be considered as a change in biological condition beyond the normal resting state that challenges homeostasis and, thus, presents a threat to the fish's health. These stress-induced changes are grouped as primary; secondary, which includes metabolic, hematological, hydromineral, and structural; and tertiary or ‘whole animal’ responses. Many of these responses can be used as quantitative indicators of stress although investigators need to be aware of the various ‘nonstress’ factors that can also influence these conditions. A major focus of current research is on the response of the hypothalamic-pituitary-interrenal axis and the resultant elevation of circulating corticosteroids. Stress, through the action of corticosteroids, may (a) reduce immunocompetence by influencing lymphocyte numbers and antibody-production capacity, and (b) affect reproduction by altering levels and patterns of reproductive hormones that influence maturation. Stress may also alter metabolic scope in fish and affect growth, partly as a result of the catabolic or gluconeogenic effect of corticosteroids. Although certain stressors encountered during normal aquacultural procedures may be unavoidable, a number of practical approaches are suggested that would help to alleviate the detrimental effects of stress in fish.