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Egypt. Poult. Sci. Vol (37)(II): (491-504)(2017) (1729)
Egyptian Poultry Science Journal
http://www.epsaegypt.com
ISSN: 1110-5623 (Print) – 2090-0570 (Online)
BIOCHEMICAL AND HISTOLOGICAL ALTERATIONS IN LIVER AND
KIDNEY OF GROWING RABBITS FED ON OVERABUNDANT DRIED
CITRUS LIMON
Z. S. H. Ismail1; H. A. M. Elwan2
1Anim. Prod. Dep., Fac. of Agric., South Valley Uni., Egypt
2Anim. Prod. Dep., Fac. of Agric., Minia Uni., Egypt
Corresponding author: Dr. Hamada Abdel-Hameid Mehany; E-mail: hamadaelwan83@mu.edu.eg
Received:14/04/2017
Accepted:15/05/2017
ABSTRACT: Fruit and vegetables are under continuous analysis worldwide to figure out or
detect new treatments to animal and human diseases. This study was intended to examine effects
of dried C. limon (DCL) on New Zealand White rabbits liver and kidney functions of 10 and 20g
DCL/kg diet. The rabbits were randomly allocated into three treatments groups of control and
two dried Citrus limon groups (10 and 20 g DCL/kg diet). Feed and water were served ad-libitum
throughout the 8 weeks of experimental period. At the end of the administration, there were a
significantly increment of GOT, GGT and ALP activities. Contrary, total bilirubin was
significant (p<0.01) decreased with feeding growing rabbit on DCL levels. While, there were no
significant differences (p>0.05) among all dietary treatments on GPT, LDH, BA and CB values;
substantial intension in KC and P levels were noticed as related to DCL supplementation.
Furthermore, the values of creatinine, urea, uric acid, Ca, K, Na and Cl had no differences
(p>0.05) among all groups. Also, serum total protein, albumin and globulin levels had the same
trend. Likewise, the histological results of liver and kidneys revealed no cellular abnormality in
the entire treatment groups as compared to control group. Generally, it is shown that DCLcan be
included up to 20 g/kg in rabbit diets without any adverse or deleterious effect on the
histomorphology of the liver and kidney of rabbits, as it stimulates the cells architecture thereby
preserving and increasing the cellular profile.
In conclusion, these results suggest that the addition of DCL to the growing rabbit’s diet could
significantly enhance liver and kidney function.
Key words: Citrus limon – Rabbit – Histology – Biochemical.
Z. S. H. Ismail; H. A. M. Elwan
492
INTRODUCTION
The effect of fruit and vegetables on
performance, physiological response and
antioxidant status of animals is currently a
topic of great scientific and practical
importance, due to the trend in reducing the
use of antibiotics in feed as well as in therapy
(Yang et al., 2009). Studies on the plant
products efficacy have given conflicting
results (Huyghebaert et al., 2011), and this is
a stimulus to implement research activities in
this direction. Lemon trees are fruit trees that
produce small, oval, yellow citrus fruits.
Citrus limonis is the scientific name of it. The
lemons are a type of berry called a
hesperidium. Lemons can be used in various
ways in our life. They can be used as
preventative and domestic medicine because
they are very rich in vitamin C,
bioflavonoids, acids and volatile oils that
help the body fight infections. Since they
are very acidic, they cause an alkalizing
effect upon the body. The sweetened juice
relieves gingivitis, stomatitis, and
inflammation of the tongue. Flavonoids are
widely dispersed group of polyphenol
compounds with health-related characteristics
that are based on their antioxidant activity,
e.g. inhibition of platelet aggregation,
anticancer, anti-inflammatory and antiviral
activities. The flavonoids present in Citrus are
Flavanones, flavones, and flavonols, although
flavones and flavonols are found in low
concentrations as compared to flavanones, but
are potent antioxidants and free radical
scavengers (Armando et al., 1998). It has been
exhibited that Citrus fruit have highest
antioxidant activity (Chun et al., 2008), due to
the presence of abundant flavonoids, vitamin
C and carotenoids (Xu et al., 2008). Common
species of the genus Citrus are Citrus indica,
Citrus aurantifolia, and Citrus limon, in
which Citrus limonis available in Egypt and
commonly known as lemon. It has significant
economic value for its essential oil and is
reported to be the source of magnesium,
potassium, vitamin C, folic acid, limonoids
and flavonoids (Deyhim et al., 2008). Citrus
limonhas shown usefulness as antidote against
certain venom, due to its platelet inhibitory
effect (Arias and Ramon, 2005), however it
needs further confirmation. More attention
has also been paid on antioxidant capacity of
Citrus limon (Berhow and Smolensky, 1995;
Xu et al., 2008), since increase dietary
antioxidant constituents could help to prevent
diseases (Hernandez et al., 2009; Gonzalez et
al., 2010). Citrus limon benefits are due to its
wide range of bioflavonoids, including rutin,
hesperidin, quercitrin, eriocitrin, narirutin,
didymin and naringin (Nijveldt et al., 2001;
Tripoli et al., 2007). Two more isomers of
hesperidin, neohesperidin and homoeriodicty
olrutinoside have also been identified in
Citrus limon (Gonzalez et al., 2010). Blood
biochemical and histological changes are
reflections of the effects of dietary treatments
on animals in terms of the type and amount of
feed ingested and available for the animal to
meet its physiological, metabolically
necessities (Ewuola et al., 2004). Thus,
dietary components have measurable effects
on blood components; hence blood
constituents are widely used in nutritional
evaluation, and survey of animals (Olorode et
al., 1995). Farm animals are being fed by
human with forages, Citrus limon which
may contain certain elements and metals that
can have adverse effect on the wellbeing of
the animal and in turn influence the
digestive/metabolic life of the animals
negatively. Glutamic Oxaloacetic
Transaminase (GOT) is found in appreciable
quantities in the heart, pancreas, muscle, and
liver. It is the serum enzyme that reflects the
functionality of heart and liver. Glutamic
Pyruvic Transaminase (GPT) is found
principally in the liver and in many tissues,
Citrus limon – Rabbit – Histology – Biochemical.
493
thus it is of limited use for rabbit disease
(Jekins, 2000). When there is liver cell
damage, the serum or plasma level of both
GOT and GPT rises tremendously. Thus, GPT
is more specific for detecting liver
inflammation, liver cell damage, and necrosis
such as that caused by parasites and hepatic
lipidosis (Jekins, 2000). Alkaline phosphatase
(ALP) is also found in appreciable amounts
especially in the liver, small intestines,
kidney, placental tissues, and osteoblasts; the
organs usually release the enzyme into the
blood. Some ALPs are normally excreted in
the bile. However, according to Ravel (1995),
ALP is found in many tissues in the body, with
the highest concentrations found in the liver,
biliary tract, epithelium, bone, and intestinal
mucosa. GOT is found in several organs and
tissues (for example, liver, skeletal muscles,
heart, and red blood cells). ALT is
predominantly found in the liver, with a
moderate-sized component in the kidney, and
small quantities in the heart and skeletal
muscles. When rabbits become sick with
suspected liver disease(s), chemistry panel
(part of the blood work) is performed to
determine liver disease that caused elevated
levels of GOT, GPT and ALP (Meredith and
Rayment, 2000). Blood biochemistry could be
used to monitor the progress of disease before
final evaluation by pathology of arteries and
organs (Aguilera et al., 2002; Marinou et al.,
2010; Tsantila et al., 2010). Albumin is a
protein specifically made by the liver and is
easily and cheaply analyzed in the laboratory.
It is mainly constituted of 60 % total protein.
Decreased levels of albumin are found in
chronic liver disease (cirrhosis) and also
nephritic syndrome where it is lost in urine.
Low albumin results to oedema since the
intravascular oncotic pressure is higher than
the pressure in extravascular space. Albumin
diffuses through damaged membrane and is
filtered by the kidney because of its molecular
size of 65000 kD (Monica, 2009). Total
bilirubin refers to both unconjugated and
conjugated bilirubin. Conditions which,
through hemolytic processes, produce
bilirubin faster than the liver can metabolize
it, cause the levels of unconjugated (indirect)
bilirubin to increase in the circulation. Liver
immaturity and several diseases, in which the
bilirubin conjugation is impaired, cause
similar elevations of circulating unconjugated
bilirubin. Bile tract obstruction or damage to
hepatocellular structure also causes increases
in levels of both direct and indirect bilirubin
in the circulation (Balisteri and Shaw, 1987).
In the liver, bilirubin is conjugated with
glucoronic acid for solubilization and
subsequent transport through the bile duct and
elimination via the digestive tract. An increase
in conjugated bilirubin is highly specific for
disease of the liver or bile ducts.
Hepatocellular injury or cholestasis is
suspected when more than 50% of total
bilirubin is conjugated bilirubin (Fody, 2005).
Rabbits are one of the most important farm
animals, which produces meat at 10-15 times
or more its own weight in a year through
progenies. They produce high class protein
characterized as lean meat and excellent fur.
The primary function of the kidney is the
formation of urine. The kidney performs
several functions which aid maintains
physiological integrity of the extracellular
fluid volume. These processes are:
conservation of water, fixed cations, glucose,
and amino acids; conservation being used in
the broad sense to imply the return to the body
fluids of the amount of the substance required
by the body (Kluwe, 2001). Elevations in
blood urea nitrogen (BUN) concentration are
seen in inadequate renal perfusion, shock,
diminished blood volume (prerenal causes),
chronic nephritis, nephlosclerosis, tubular
necrosis, glomerular-nephritis (renal causes)
and urinary tract obstruction (postrenal
causes). Urea is used to detect any
abnormality of the kidneys. Urea nitrogen in
Z. S. H. Ismail; H. A. M. Elwan
494
plasma and serum levels is shown to be higher
in men than women (Kamath et al., 2001).
Thus, the applications of laboratory tests can
be used to evaluate the functional status of
several organs notably the liver and kidneys.
This study was aimed to examine the
suitability of using dried Citrus limon in diets
of growing rabbits to ascertain its suitability
as an additive, and its effects on the liver and
kidney function.
MATERIALS AND METHODS
Experimental animals
The experimental work was carried out at the
farm of Animal and Poultry Production
Department, Faculty of Agriculture, Minia
University in a semi closed housed on
galvanized wire cages (40 × 50 × 35 cm)
provided with feeders and automatic drinking
watery system, and were kept under the same
managerial, hygienic and environmental
conditions. A period of 14-16 hours of day
light was provided. Feed and water were
available all time ad libitum during the
experimental period (8 weeks).
Preparation of dry lemon
Lemonwas provided from a private
commercial market at El-Minya Governorate
Egypt. The lemon was dried at 40ºC until
constant weight. The dry lemon was finally
milled, sieved (1 mm mesh) and stored in a
well tight polyethylene bags at room
temperature 25°C. Composite sample of
lemon powder was taken in sample plastic bag
for nutritional analysis.
Rabbitsdiet and lemon Supplementation
Three batchesofrabbitsdiet each of 500 kg
were formulated to contain; 44% ground
yellow corn, 40.5% wheat bran, 13.5%
soybean meal (44% crude protein), 0.5% lime
stone, 1% sodium chloride and 0.5% vitamin
& mineral premix. Citrus limon powder was
added and thoroughly hand mixed with other
feed ingredients of each batch at 0, 10 and 20g
DCL/kg diet. Experimental diets were packed
in polyethylene bags until feeding.
Animals and treatments
Forty eight unsexed growing New Zealand
White (NZW) rabbits aged eight weeks
weighed in average 1543.33 ± 25g were
randomly blocked by weight into three groups
(16 animals each), where the 1st group fed a
basal ration free of dried Citrus limon
(control), while the 2nd and 3rd groups were
fed on the same basal ration supplemented
with 10g and 20g DCL/ kg diet respectively.
The experiment lasted to 16 weeks of age.
Blood sampling
At the end of experimental period (8 to 16
weeks of age) 5ml of blood samples were
collected in non-heparinized tubes from one
animal each replicate, then allowed to clot at
room temperature. Serum was separated by
centrifugation at 3000 rpm for 10 minutes and
stored at -20o Cuntil analysis of liver and
kidney functions.
Liver Function Tests (LFTs)
Estimation of liver functions by measuring,
the activities of liver enzymes such as GOT,
GPT, ALP, LDH, GGT, BA, TB and DB
using diagnostic kits (Vitro, Germany). Also,
total protein, albumin, globulin, albumin
globulin ratio were determined using
commercial kits (Bio-Med, Egypt), where,
alanine and aspartate aminotransferases were
determined based on the colorimetric
measurement of hydrazone formed with 2, 4
dinitrophenyl hydrazine (Reitman and
Frankel, 1957), Gamma glutamyltransferase
activity was determined by the method of
Rosalki et al. (1970) using gamma -glutamyl-
p-nitroanilide as substrate, alkaline
phosphatase by the phenolphthalein
monophosphate method (Babson, 1965). The
activity of LDH was assayed by the method of
Horecker and Kornberg (1948), total protein
was determined by the Biuret method (Peters,
1968), albumin by the bromocresol green
Citrus limon – Rabbit – Histology – Biochemical.
495
method (Doumas et al., 1971), total bilirubin
and direct bilirubin concentrations (Fischbach
et al., 1996). All analytical testes were done
using T80 UV Spectrophotometer UK, at
Animal and Poultry Production Department,
Faculty of Agriculture, Minia University.
Kidney Function Tests (KFTs)
The serum was analyzed for levels of urea,
uric acid, creatinine, creatinine kinase,
calcium, phosphate, potassium, sodium and
chlorine and results were expressed as mg/dL,
U/L and mmol/L respectively, using
commercially Bio-Med reagent kits Egypt
according to determined according to (Tietz,
1986, 1990). While, blood electrolytes were
determined using Vet scan VS2 Blood
chemistry and electrolyte analyzer (ABAXIS-
Germany) at Animal and Poultry Production
Department, Faculty of Agriculture, Minia
University.
Histopathological Examinations
For the histopathological analysis, the
tissue samples from liver and kidney were
collected from the slaughtered rabbits (8
rabbits per treatment) at the end of
experimental period (8 to 16 weeks of age)
and fixed in 10% BPS formalin saline buffer.
Tissues were dehydrated by immersing
through a series of alcohols of increasing
concentrations (from 70% to absolute),
infiltrated with xylene, and embedded in
paraffin. The casting of blocks was carried
out in stainless block models. The rotary type
fully automated microtome (model FAM 47,
Acculab, USA) was used for cutting the
paraffin sections. Then blocks were trimmed,
and sections of 3 mm thickness. Six to seven
inches continuous long ribbons of the material
were cut and laid on constant temperature
water bath (around 50°C). The sections were
separated with a heated scalpel after they
spread completely. The cut sections were
mounted on the clean glass slides using
Mayer’s egg albumin as the section
adhesive. The mounted slides were dried in
paraffin oven at 58-60°C for 1 h. The tissue
sections were stained by the Harris
hematoxylin and eosin staining method using
fully automated staining machine (model LST
94, Hestion, Australia). The paraffin sections
were deparaffinized with the xylene before
hydration through graded alcohol to distilled
water. This was followed by the dehydration
in ascending grades of alcohol. The clearing
was performed in the xylene, and a drop of
distrene plasticizer xylene mountant was
placed on a coverslip and the section on the
slide pressed on it. The slide was inverted, and
the cover slip was pressed with a rod to
remove the air bubbles if any trapped.
Sections were examined at a magnification of
×40 under a LED Fluorescence microscope
fitted with the stage micrometer. Microscope;
photomicrographs of the samples were
recorded using an Optika Research LED
Fluorescence Microscope (model B-
500TIFL) (Diab et al., 2012).
Statistical Analysis
The study was conducted based on a
completely randomized design (CRD) with
three treatments and 16 replicates per
treatment 1 animal each. Data were analyzed
by Statistical Analysis System software (SAS,
Version 9.1.3, 2003) using the generalized
linear model (GLM) procedure. However,
significant differences among treatment
means for each trait in experiment was
detected using Duncan’s multiple rang test
(1955). The statistical model was as follow:
Xij= µ + Ti + Eij
Where; Xij = value observed in each
experimental unit, µ = mean population, Ti =
the effect of each treatment, and Eij =the effect
of experimental errors.
RESULTS AND DISCUSSION
Liver function
The obtained data revealed that, no significant
difference (p>0.05) between dietary
treatments was observed in the values of GPT,
LDH, BA, DB, TP, albumin, globulin and
Z. S. H. Ismail; H. A. M. Elwan
496
AGR (Table 1). The non-significant
difference may be due to the biological
activities of phenolic compounds present in
the additive which may have contribution to
the enhancement of the rabbit health status.
Even though, as shown in Table 1, there are
significant differences (p<0.05) between
dietary treatments were observed in the mean
values of GOT, ALP, GGT and TB. Adding
DCL supplementation to growing rabbits
depicted increased enzyme levels with
increasing DCL level. The significant
differences (p<0.05) observed in the GOT,
ALP, GGT and TB still remains within the
normal values as obtained by Ewuola and
Egbunike (2008), and Ezenwanne and Ucheya
(2012), in clinically healthy rabbits. The value
of the serum enzymes was in line with earlier
report of Ewuola and Egbunike (2008); and
Ezenwanne and Ucheya (2012). The results of
the histological examination of the liver (Fig.
2) did not show any visible alteration in the
liver cells among all treated groups. These
results may give approve the critical role of
flavonoids present in Citrus which that are
based on their antioxidant activity, e.g.
inhibition of platelet aggregation, anticancer,
anti-inflammatory and antiviral activities. The
flavonoids are Flavanones, flavones, and
flavonols, although flavones and flavonols are
found in low concentrations as compared to
flavanones, but are potent antioxidants and
free radical scavengers (Armando et al.,
1998). In addition, it has been exhibited that
citrus fruit have highest antioxidant activity
(Chun et al., 2008), due to the presence of
abundant flavonoids, vitamin C and
carotenoids (Xu et al., 2008).
Kidney function
The effect of dietary dry citrus limon on
kidney function of growing rabbits such as
creatinine kinase, creatinine, urea, uric acid,
Ca, P, K, Na and Cl are shown in Table 2; the
data revealed that adding DCL to growing
rabbit diets at all levels recorded a significant
improvement (p<0.05) in the values of C K
and P. However, no significant (p≥0.05)
alleviation was detected in the values of
Creatinine, Urea, Uric Acid, K, Na, and Cl as
a resulted of the previous addition. The result
from this study demonstrates that C. limon
treatment does not induce electrolyte
imbalance in treated rabbits. Sodium is the
most abundant extracellular ion, and it plays
an important role in muscle contraction.
Similarly, potassium, an abundant
intracellular ion, plays a vital role in muscle
contraction. The results of electrolyte levels
were in a normal range of rabbits resulting
from the serum level of sodium seen in this
study thus provides evidence that the use of
citrus limon dose not present risk for renal
function in the experimental groups that the
sodium and potassium levels were not
significant when compared to the control
groups. Creatinine and urea concentration
were extremely insignificant in all the treated
groups as compared to the control group at
(p<0.01) as represented in Table 2. Hence, an
abnormally elevated blood creatinine is
diagnostic of impaired renal function (Henry,
2001; and Wallech, 2009). Moreover, the
result reviled that administration of C. limon
to rabbits had a positive effect on the kidney
in accordance with Solomon et al. (2015) who
concluded that, administration of C. limon
juice to rabbits has a positive effect on the
kidney and added that, it could be suggested
that the breeder should supplement animal
feed with this beneficial fruit as it will help
improve animal health and prevent any
incidence of kidney malfunctioning. Also,
Manners (2007) reported that citric acid
present in C. limon helps in dissolving kidney
stones and Benavente-Garcia et al. (2007)
reported that ascorbic acid present in C. limon
is a natural antioxidant. It also showed that
lemon juice has an effect on the level of uric
Citrus limon – Rabbit – Histology – Biochemical.
497
acid in the kidney. Kang et al. (2007) reported
that when lemon has been fully metabolized
in the body, the pH of the body is raised. This
implies that reduced level of uric acid
improves kidney function. The microscopic
appearance of the kidneys has shown that
supplementation with 10 and 20g DCL/kg diet
revealed normal cellular profile, no
abnormality seen as control group and this
confirm the results of kidney function tests,
there were no adverse effects of excises dry
citrus limon up to 20g/kg diet.
Histological alterations
The basic structure of liver sections after 8
weeks of dietary treatment, showed numerous
hepatic lobules (Figure 2). The central vein is
located in the middle of the lobule. The
hepatocytes are polygonal in shape with
granulated, eosinophilic cytoplasm and
centrally located nuclei with one or two
nucleoli and delicate strands of chromatin.
Also, Kupffer cells appeared between the
hepatocytes as spindle-shaped cells Fig. 2 (A,
B and C). These cells act as an effective
particulate filtration system in the liver tissue
to prevent bacteria and other foreign materials
to penetrate from the central vein to the
systemic circulation .This is due mainly to the
phagocytic activity of Kupffer cells, which
might explain the better immunity of treated
compared to control rabbits in terms of higher
plasma globulin level. Moreover, the kidney
consists of an outer cortex and an inner
medulla. The outer cortex of control kidneys
contains the renal corpuscles which appear as
large spherical structure and renal tubules
(proximal and distal convoluted tubules).
Each renal corpuscle is surrounded by the
Bowman's capsule composed of simple
squamous epithelial cells. It encloses the
urinary space and the capillary tuft of the
glomerulus which consists of blood capillaries
Fig. 2 (D, E and F). The previous finding may
insure that the citrus fruit have highest
antioxidant activity (Chun et al., 2008), due to
the presence of abundant flavonoids, vitamin
C and carotenoids (Xu et al., 2008).
CONCLUSION
The results obtained from this study show that
dried citrus limon levels do not disrupt the
activities of the liver and kidneys. This
indicates its safe usage as growth additive in
growing rabbits.
498
Z. S. H. Ismail; H. A. M. Elwan
Table (1): Liver function alteration of New Zealand white rabbits fed on 0, 10 and 20g/kg diet of dry Citrus limon
a, b and c values within a column with different superscripts differ significantly at p<0.05, SEM: Standard error of the mean. P= P; DCL= Dried
Citrus limon; GPT= alanine aminotransferase; GOT= aspartate aminotransferase; ALP= alkaline phosphatase, LDH= lactate dehydrogenase; GGT=
γ-glutamyltransferase; BA=bill acid; TB=total bilirubin and DB= direct bilirubin.
Table (2): Kidney function alteration of New Zealand white rabbits fed on 0, 10 and 20g/kg diet of dry Citrus limon
Treatments
CK
(U/L)
Creatinine
(mg/dl)
Urea
(mg/dl)
UA
(mg/dl)
Ca
(mmol/L)
P
(mmol/L)
K
(mmol/L)
Na
(mmol/L)
Cl
(mmol/L)
Control
158.20c
2.01
51.23
3.39
3.63
1.20b
4.53
144.00
103.00
DCL10g/kgdiet
175.73b
2.252
52.82
3.36
3.53
1.50a
4.46
144.66
101.00
DCL20g/kgdiet
198.33a
2.23
53.84
3.25
3.83
1.60a
4.30
145.33
103.66
±SEM
6.88
0.28
2.43
0.08
0.13
0.10
0.10
1.81
0.69
P Value
0.01*
0.53NS
0.75NS
0.52 NS
0.35NS
0.02*
0.31NS
0.87NS
0.07 NS
a, b and c Values within a column with different superscripts differ significantly at p<0.05, SEM: Standard error of the mean. P= P value; DCL=
Dried Citrus limon; CK= Creatinine Kinase; Ca= calcium; P= phosphorus, K= potassium; Na= sodium; Cl=chloride and UA=Uric Acid.
Treatments
GPT
(U/L)
GOT (U/L)
ALP
(U/L)
LDH
(U/L)
GGT
(U/L)
BA
(µmol/L)
TB (mg/dl)
DB
(mg/dl)
Control
7.266
11.093b
10.100b
140.256
23.000b
3.560
0.736a
0.233
DCL 10g/kg diet
7.333
11.533a
17.150a
139.428
26.000a
3.553
0.626b
0.223
DCL 20g/kg diet
7.466
11.933a
26.933a
138.153
25.333a
3.583
0.596b
0.220
±SEM
0.074
0.093
3.504
17.360
0.693
0.055
0.014
0.011
P value
0.234NS
0.001**
0.036*
0.116NS
0.049*
0.922NS
0.001**
0.704NS
Citrus limon – Rabbit – Histology – Biochemical.
499
Figure (1): Protein profile of New Zealand white rabbit’s serum as affected by dietary 0, 10
and 20g/ kg diet of dry Citrus limon.
Z. S. H. Ismail; H. A. M. Elwan
500
Figure (2): Photomicrograph of liver (A, B and C) and kidney (D, E and F) sections showing
normal histological structure stained with general stain (H&E)
A) Control
B) 10g DCL/ kg diet
C)20g DCL/ kg diet
D)Control
E) 10g DCL/ kg diet
F) 20g DCL/ kg diet
Citrus limon – Rabbit – Histology – Biochemical.
501
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1
2
1
2
2
GOT, GPT, ALP, GGT, LDH and CK
p>0.05
GOT ALP CK
