Basrah Journal of Veterinary Research,Vol.15, No.3,2016
Proceeding of 5th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
34
STUDY THE EFFECT OF DIFFERENT LEVEL OF
FRUCTOOLIGOSACCHARIDE (FOS) ON SOME BLOOD INDICES IN
YOUNG COMMON CARP (CYPRINUS CARPIO L.)
Nasreen M. Abdulrahman*,Vian M. Ahmed**, Hawkar J. Hama Ameen***
Bakhan Rafiq Hasan****
*,**, ****Department of Animal Production, Faculty of Agricultural Sciences, University of
Sulaimaniya, Iraq
***Department of Biology, School of Sciences, University of Raparin, Iraq
Keywords: Prebiotic FOS, Blood indices, Common carp.
ABSTRACT
The present study was conducted to investigate the effects of dietary
fructooligosaccharide (FOS) on some blood indices of common carp Cyprinus carpio. The
fingerlings were adopted for 2 weeks and then reared in triplicate groups in 9 tanks (four fish
per tank with average initial weights of (40 ± 3 g). Fish fed experimental diets containing
different levels of fructooligosaccharide (FOS); (7.5 and 10 g FOS/kg diet) to apparent
satiation twice a day for 10 weeks. The results showed significantly differences in Red blood
cell count (1012 cells/dl) in T2. Hemoglobin (g/dl) data was 11.597 in the control. The mean
corpuscular hemoglobin (pg/cell) was 63.333 in control, 56.033 in T21 and 52.100 in T2;
mean corpuscular hemoglobin concentration (g/l) was 28.100 in the T1; mean corpuscular
volume were (fL) 215.950 in control. White blood cell (109 cells/dl) data was (112.350) for
control; granulocyte % was (15.050) in control; Lymphocyte % was (82.750) for control;
Platelets % was (56.500) in T2 and monocyte % was (13.300) in T1.
INTRODUCTION
Aquatic animal production is the industry sector with the highest growth among animal
production sectors. Between 1970 and 2010 it had an average annual growth rate of 2.9%,
while total terrestrial meat production had an average annual growth rate of 2.7% [25].
Common carp Cyprinus carpio L. is one of the most important farmed species in world
aqua culture, especially in Asia. It was ranked in the third place (3,043,712 tonne) among all
freshwater species worldwide in 2005, more than 90% of which comes from Asia [11].
Improving the health conditions and growth performance in commonly farmed fish such as
Basrah Journal of Veterinary Research,Vol.15, No.3,2016
Proceeding of 5th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
35
common carp is a topic of strenuous interest. Hence, such components as antibiotics,
hormones, ionophores and some salts have been frequently used to enhance fish health and
growth [16].
Prebiotics mainly consist of oligosaccharides promoting beneficial
bacterial growth within the gastrointestinal tract of higher vertebrates
[13]. In different studies from last ten years, many substances have been investigated as
prebiotics. Based on the studies of [18] any food stuff that reaches the colon (e.g. nondigestible
carbohydrates, some peptides and proteins, as well as certain lipids) is a candidate
prebiotic [27].
However, most studies have focused on non-digestible carbohydrates, mainly
oligosaccharides. According to [12] only three oligosaccharides were classified as prebiotics:
inulin, transgalactooligosaccharide (TOS) and lactulose. A more recent study includes
fructooligosaccharides (FOS) in the list of prebiotics [21]. Mannanoligosaccharides (MOS)
and fuctooligosaccharides (FOS, or oligofructose) are perhaps the most well studied
oligosaccharides in fish but some information regarding applications of
galactooligosaccharides, xylooligosaccharides, rabinoxylooligosaccharides and
maltooligosaccharides in aquatic animals is also available [9].
Among prebiotics, fructooligosaccharides (FOS) is nondigestible
carbohydrates, which selectively stimulate the growth and metabolism of health promoting
bacteria present in the host gut. It is reported that FOS could overcome the limitations and
side effects of antibiotics and other drugs, leading to high production through enhanced
growth, stimulated immune response and increased
resistance to pathogens of fish [14]. Previous studies also demonstrated the efficacy of FOS
to increase the growth performance and the non-specific immunity of fish [28]. However,
some researchers have proved that continuous administration of high levels of
immunostimulants has no beneficial effects on growth or/ and immunity [5]. Others also
suggested that discontinuous administration of immune stimulants may solve those problems
[7]. Unfortunately, up to date, there were no report concerning the feeding modes of FOS on
growth, immunity and disease resistant in fish. So the main objectives of the present studies
evaluate the effect of adding FOS as prebiotic on common carp blood indices such as RBC
(Red Blood Cell; 10 12 cells/l); WBC (White Blood Cell; 109cells/l); Hb (Hemoglobin; g/l);
Basrah Journal of Veterinary Research,Vol.15, No.3,2016
Proceeding of 5th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
36
MCH (Mean Corpuscular Hemoglobin; pg); MCHC ( Mean Corpuscular Hemoglobin
Concentration; g/l); MCV ( Mean Corpuscular Volume; fL), white blood cell (WBC) ,
granulocyte %, Lymphocyte %, Platelets %, monocyte %.
MATERIALS AND METHODS
The experiment was conducted for 70 days we used 36 young common carp C. carpio
L. The fish were distributed among experimental tanks with mean initial weight of (40 ± 3 g).
They were pre-acclimated to laboratory conditions for 14 days prior to the feeding trials.
Each tank was provided with a proper continuous aeration. Each tank was stocked with four
fish. In control fish were fed an experimental diet without FOS, While in T1, fish were fed a
diet 7.5 g/kg FOS, T2 represents the third treatment, in which fish were fed on a diet 10 g/kg
FOS.
Diet formulation: Experimental diets were prepared with Animal concentrate Soya meal,
Yellow corn, Wheat bran, Barely and Premix, and the composition of the different diet shown
in Table (1). The ingredients were mixed with water to obtain dough. Then, the dough was
passed through an electrical mincer for pelleting by using Kenwood Multi-processors. The
pellets were dried at room temperature for a few days and crushed to yield fine particles. The
fish were fed 2 times a day, once was at 9:00 am and another time at 2:00 pm. Feeding rate
started with 4% of biomass then the accurate feeding rate was determined to be 3% by third
week depending on satiation level. Fish were individually weighed bimonthly. The feeding
amount was then recalculated according to new weights.
Basrah Journal of Veterinary Research,Vol.15, No.3,2016
Proceeding of 5th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
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Table 1: percentage of the feed ingredients in fish diets:
At the end of the experimental period, three fish were randomly taken from each
experimental group. All fish samples were weighed individually. The blood samples from
each fish of the different groups were collected by cutting of the caudal peduncle. Whole
blood samples were collected in small plastic vials containing heparin for determination of
some blood parameter and the concentrations were determined by using the hematology
analyser BC-2800 is a compact, fully automatic hematology analyser with 19 parameters for
complete blood count (CBC) test. RBC (Red Blood Cell; 10 12 cells/l); WBC (White Blood
Cell; 109cells/dl); Hb (Hemoglobin; g/dl); MCH (Mean Corpuscular Hemoglobin; pg);
MCHC ( Mean Corpuscular Hemoglobin Concentration; g/l); MCV ( Mean Corpuscular
Feed ingredients Percent
Animal concentrate 10
Soya meal 40
Yellow corn 15
Wheat bran 18
Barely 15
Premix 2
Total 100
Chemical Composition
Crude protein % 27.351
Crude fat % 2.584
Crude fiber % 6.155
Energy KGal/ kg 2235.2
Ash % 87.61
Basrah Journal of Veterinary Research,Vol.15, No.3,2016
Proceeding of 5th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
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Volume; fL), (WBC) white blood cell(109 cells/dl) , Granulocyte %, Lymphocyte %, Platelets
%, Monocyte %.
Statistical analysis: The experimental was conducted using the (CRD) design and the
general linear models (GLM) procedure of XLSTAT. Pro. 7.5 one way (ANOVA). Duncan
test’s was used to compare between means of the control and experiment treatments. The
mode of analysis was as follows: [9]
Yij = μ + Ti + Eij μ = The overall mean.
Ti = The effect of treatment. Eij= The random error.
RESULTS AND DISCUSSION
Table (2) summarizes the blood parameters results for the date variety used in this
study. The data indicate that FOS affects significantly the Red Blood Cell (RBC) in control
(1.633), Hemoglobin (g/dl) was 11.133, 10.633 and 9.735 for the T1, T2 and control
respectively, observed that control significantly (P≤0.05) differ. The Mean Corpuscular
Hemoglobin (pg/cell) data in Table (2) were 63.333, 56.033 and 52.100 for control, T1 and
T3 and the significant (P≤0.05) different among treatments appeared in control which was
significantly higher than other treatments. The Mean Corpuscular Hemoglobin Concentration
(g/l) were 28.100, 25.900 and 25.350 for the T1, control and T2 respectively, and the T1 was
higher than control and T2 significantly (P>0.05). Mean corpuscular volume (MCV) ranged
213.800, 210.900 and 208.867 for the control, T2 and T1 respectively.
Table 2: Effect of FOS on some blood parameters of common carp during 70 day
of rearing:
T
RBC
(1012 cells/dl)
Hb
(g/dl)
MCH
(pg/cell)
MCHC
(g/l)
MCV
(fL)
Control 1.633c ± 0.024 9.735c ± 0.145 63.333a ± 1.615
25.900b ±
0.200
215.950a ±0.150
T1 1.757b ± 0.013
11.133a ±
0.133
56.033b ± 0.825 28.100a ± 0.600 208.867b ± 0.561
T2 1.947a ± 0.003 10.633a ±
0.517
52.100c ± 0.252 25.350b ± 0.350 210.900c ± 0.503
Mean values with different superscripts within a column differ significantly (P≤0.05).
Basrah Journal of Veterinary Research,Vol.15, No.3,2016
Proceeding of 5th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
39
The present results agree with that of [4] they used prebiotic Immunogen
(supplementation at concentrations of 5 and 10 g kg–1 prebiotic) in RBC, and with [1], in
which the level of 2.5 g/kg Fructooligosaccharide affect significantly the RBC, WBC and
Hemoglobin. Their results, together with ours, indicate that research in this area should
continue and causal relationships should be sought between dietary prebiotics and some
haematological and serum plasma biochemical parameters of fish [20], in this study, RBC
and MCH did not change after feeding with inulin.
The adding of FOS has different stimulation in present results as shown in (Table, 2).
This means that a stimulation of the immune response of fish through dietary supplements is
possible and is of high interest for commercial aquaculture as stated by [24]. The innate
immune system is very important in this regard because aquatic animals are continually
vulnerable to numerous opportunistic pathogens and this part of immune response provides
the first line of defence for the host [17]. The results of the study of [23] showed that dietary
FOS could modulate the innate immune responses of Caspian roach fry. The
immunostimulatory nature of prebiotics may be attributed to stimulation of the growth of
beneficial bacteria such as lactic acid bacteria and Bacillus spp. [22], which possess cell wall
components such as lipopolysaccharides which have immunostimulatory properties [26].
However, [6] reported that dietary inulin (5 or 10 g inulin /kg) had no effect on the innate
immune response of gilthead seabream (Sparus aurata L.) compared to the control group (0 g
/kg). This contradictory result may be attributable to the low dosage, different duration of
prebiotic administration, life stage and/or different fish species [19].
White blood cell (WBC) in Table (3) presented 112.350, 109.000 and 105.650 for T1,
T2 and control respectively, where control was significantly differ than other treatments. The
data as presented in Table (3) the granulocyte percentage showed 15.050, 12.450 and 7.750
for T12, T2 and control respectively, and observed that T1 was significantly higher than
control and T2 (P>0.05). In Table (3) Lymphocyte percentage were 82.750, 75.450 and
73.980 for control, T2 and T3 respectively, observed control had significant (P≤0.05)
different among other treatments. In the same table monocyte percentage showed 13.300,
12.200 and 11.850 for T1, T2 and control respectively, control was significantly lower than
other treatment. Table (3) showed and the platelets percentages were 56.500, 53.000 and
31.000 for the T2, T1 and control respectively.
Basrah Journal of Veterinary Research,Vol.15, No.3,2016
Proceeding of 5th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
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Table 3: Effect of FOS on on differential WBC count of common carp during 70
day of rearing
T
WBC
(109cells/l)
GRAN % Lymph% Mon% PLT%
contro 105.650c ± 0.800 7.750c ± 0.350 82.750a ± 1.050 11.850b ± 0.450 31.000B ± 1.00
T1 112.350a ± 0.150 15.050a ± 0.050 73.950b ± 0.350 13.300a ± 0.100 53.000a ± 2.00
T2 109.000b ± 0.100 12.450b ± 0.250 75.450b ± 0.450 12.200b ± 0.208 56.500a ± 0.500
Mean values with different superscripts within a column differ significantly (P≤0.05).
Lymphocytes are one of the most important cells that can affect immune response of
fish, these cells produce antibodies by specific immunity and increase in macrophages, an
increase of such immunity cells can promote fish defences to adverse condition [2]. Common
carp fingerlings responded to the dietary prebiotic levels with significant differences (p<
0.05) in blood constituents when fed diets containing 0.5–2.5 g Immunogen/kg, one of the
most distinct effects was a rise in total protein and leucocyte levels; generally accepted that in
all vertebrates including fish, stressors elicit a stress response in leucocytes [8], the increase
in WBC count might be due to stress suffered by fish as a result of daily feeding on β-glucan.
[15] reported an increase of WBC counts in C. carpio after herbal treatment with
Azardicha indica, observed increases in the leucocyte and total protein levels as well as lower
mortalities resulting from the pathogenic A. hydrophila infection appear to be signs of
enhanced health status of the prebiotic-fed fish, the improved health condition in C. carpio
fingerlings is probably due to the β-glucan and MOS components of the Immunogen [3].
Basrah Journal of Veterinary Research,Vol.15, No.3,2016
Proceeding of 5th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
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فی بعض معاییر دم (Fructooligosaccharide) دراسة التأثیر مستوى مختلف من السابق الحیوی
(Cyprinus carpio L.) صغار أسماک الکارب الاعتیادی
نسرین محی الدین عبدالرحمن* فیان محمد أحمد** ھاوکار جعفر حمھ امین ***
باخان رفیق حسن ****
*،**،**** قسم الانتاج الحیوانی، کلیة العلوم الزراعیة، جامعة السلیمانیة، العراق
***سکول العلوم ، قسم العلوم الحیاتیة، جامعة الرابرین ، العراق
الخلاصة
أجریت ھذه الدراسة لتبیان تأثیر (فرکتولیکوسکراید) کسابق حیوی کمصدر للتکافل. اجریت التجربة فی مختبرات کلیة
الزراعة، جامعة السلیمانیة. استغرقت التجربة أشھرین بعد مدة أقلمة استمرت 15 یوم. استعمل 63 سمکة کارب اعتیادی
40 غم لاختبار تأثیر مستویات مختلفة من السابق الحیوی فی بعض معاییر دم صغار أسماک الکارب ± بمعدل وزن 3
غذیت الأسماک بعلیقة خالیة من السابق الحیوی، (control) فی المعاملة السیطرة Cyprinus carpio L. الاعتیادی
والأسماک فی المعاملة الاولى غذیت بالسابق الحیوی 7.5 غم/ کغم، غذیت أسماک المعاملة الثانیة بعلیقة 10 غم/ کغم من
السابق الحیوی تتکون کل المعاملات من ثلاثة مکررات وفی کل مکرر خمس أسماک کارب اعتیادی والتی رٌبیت فی
أحواض بلاستیکیة وتغذیتھا مرتین بالیوم. أظھرت قیاسات دم الأسماک المفحوصة فروقا معنویة تحت مستوى
فی عدد کریات الدم الحمر ( 1012 ) کریة/ لتر فی المعاملة الثانیة ( 1.947 )، سجلت بیانات الھیموغلوبین (P<0.05)
11.133 غم/ دسم) فی المعاملة السیطرة، وکان متوسط ھیموغلوبین الکریة ( 63.333 ) بایکوغرام/ کریة فی المعاملة )
السیطرة ، کان متوسط ترکیز الکریة 28.100 غم/ لتر فی المعاملة الاولى، وکان متوسط حجم الکریة 215.950
( فی عدد خلایا الدم البیض(خلیة لکل لتر 1012 (P≤ فیمتولتر فی المعاملة السیطرة ، تفوق المعاملة الاولى معنویا ( 0.05
112.350 ) ، ونسبة خلایا الدم البیض الحبیبیة کانت ( 15.050 %) فی المعاملة الاولى ، وکذلک کان نسبة الخلایا )
اللمفاویة ( 82.750 %)فی السیطرة ، الصفائح الدمویة کانت ( 56.500 %) فی المعاملة الثانیة ، و کان نسبة خلایا الدم
الوحیدة ( 13.300 %) فی الاول.
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