Adel and Khara 2014 - VC, Iron - Rainbow Trout - Growth, Hematological and Immunological Parameters
Adel and Khara 2014 - VC, Iron - Rainbow Trout - Growth, Hematological and Immunological Parameters
Adel and Khara 2014 - VC, Iron - Rainbow Trout - Growth, Hematological and Immunological Parameters
Abstract
A 60-days growth trial was conducted in a flow-through culture system to examine the
effects of different dietary vitamin C and iron levels on the growth, hematological and
immunological parameters of rainbow trout (Oncorhynchus mykiss) fingerlings (with
average initial weight of 5.12 ± 0.02 g). Three levels of vitamin C (150, 200 and 250
mg kg-1 diet) and iron (5, 7 and 9 mg kg-1 diet) (as ferrous sulfate) and their
combination were used to prepare diets used in nine experimental treatments with three
replicates. The results revealed that the supplementation of basal diet with vitamin C
and iron significantly affected the final body weight (FBW), total length (TL), feed
conversion ratio (FCR) and specific growth rate (SGR). Among the haematological
parameters haematocrit and mean corpuscular volume (MCV) were significantly
influenced by vitamin C and iron. Results also showed that lysozyme and IgM
concentrations significantly changed among dietary treatments. Based on the results of
this study, it can be conclude that addition of vitamin C and iron to the basal diet of
cultured rainbow trout will improve the growth rate and well-being of this fish.
1- Department of Fishery, Guilan Scince and Research Branch, Islamic Azad University, Rasht, Iran
2-Department of Fishery, Lahijan Branch, Islamic Azad University, lahijan, Iran, P.O.Box: 1616
*Corresponding author's Email: [email protected]
887 Adel and Khara, The effects of different dietary vitamin C and iron levels on the growth,…
mg vitamin C, T5: 200 mg vitamin C, Following that, the diets were cold-
T6: 250 mg vitamin C, T7: 5 mg Fe + pelleted into 3-mm diameter size using
150 vitamin C, T8: 7 mg Fe + 200 mg a pasta maker and then were dried in
vitamin C and T9: 9 mg Fe + 250 mg cool drier for 24 h. After drying, the
vitamin C equivalent kg-1 diet. Also, a diets were broken up and sieved into
treatment without addition of vitamin C appropriate pellet sizes, and were stored
and iron was used as control. In in a cool place until used, and then from
preparing the diet, dry ingredients were each diet one sample was taken for
first ground to small particle sizes in a chemical analysis.
mill. Ingredients were thoroughly
mixed, and then fish oil and water were
added to obtain a 25% moisture level.
a
Vitamin mixture was manually provided according to feed requirements of the fish and
ingredients were obtained from Science Laboratories (Ghazvin, Iran); which each 1000 g
vitamin mixture provides vitamin A, 1,600,000 I.U; vitamin D3, 400 000 I.U; thiamin, 6 g;
riboflavin, 8 g; niacin, 12 g; pantothenic acid, 40 g; pyridoxine, 4 g; folic acid, 2 g;
cyanocobalamin, 8 mg; vitamin C, 60 g; vitamin K3, 2 g; biotin, 240 mg and inositol, 20 g.
b
Mineral premix (mg kg)1 diet): NaCl, 500; MgSO4.7H2O, 7500; NaH2PO4.2H2O, 12 500;
KH2PO4, 15 500; Ca(H2PO4)2ئH2O, 10 000; (CH2CHCOO)2Ca.5H2O, 1650;
ZnSO4.7H2O, 176.5; MnSO4.4H2O, 81; CuSO4.5H2O, 16.5; CoCl2.6H2O, 0.53; KI, 1.59;
starch, 147.5.
889 Adel and Khara, The effects of different dietary vitamin C and iron levels on the growth,…
Table 2: Final body weight (FBW), weight gain (WG) (g/fish), total length (TL), feed conversion
ratio (FCR), specific growth rate (SGR) and condition factor (CF) of rainbow trout fed the
experimental diets for 8 weeks.
Initial Final
WG SGR (% Condition Survival
Treatments weight weight TL (cm) FCR
(%) day-1) factor rate (%)
(g) (g)
Control 5.12 ± 0.02 24.6 ± 5.3 c 27.6 ± 0.71 13.5 ± 1.08 1.5 ± 0.04 ab 2.06 ± 0.25 de 1.04 ± 0.31 100
T1 5.13 ± 0.02 25.8 ± 7.1 d 28.7 ± 1.1 14.1 ± 1.17 1.4 ± 0.03 bc 2.12 ± 0.33 d 1.04 ± 0.05 100
T2 5.14 ± 0.01 27.1 ± 3.5 e 29.7 ± 1.3 14.2 ± 1.10 1.3 ± 0.01 bcd 2.18 ± 0.15 cd 0.96 ± 0.02 100
T3 5.12 ± 0.06 28.2 ± 7.5 f 30.7 ± 1.8 14.9 ± 0.77 1.2 ± 0.02 cd 2.23 ± 0.32 c 1.04 ± 0.09 100
T4 5.15 ± 0.02 21.6 ± 4.9 a 25.3 ± 1.4 13.6 ± 0.17 1.6 ± 0.04 a 1.91 ± 0.25 f 1.03 ± 0.11 100
T5 5.10 ± 0.05 23 ± 9.9 b g 26.3 ± 1.6 13.4 ± 1.53 1.4 ± 0.04 bc 1.99 ± 0.49 e 1.06 ± 0.16 100
T6 5.11 ± 0.04 31± 7.9 b ef 33 ± 1.2 15.3 ± 0.60 1.1 ± 0.02 e 2.36 ± 0.32 a 1.12 ± 0.06 100
T7 5.12 ± 0.03 29.4 ± 4.1 d 31.5 ± 1.09 14.6 ± 0.78 1.2 ± 0.01 cd
2.28 ±0.17 b 1.13 ± 0.02 100
T8 5.13 ± 0.02 27.6 ± 1.9 d 30.1 ± 0.61 14.5 ± 0.96 1.3 ± 0.02 bcd 2.21± 0.08 c 0.93 ± 0.03 100
T9 5.13 ± 0.01 25.8 ± 3.3 c 28.7 ± 1.1 13.8 ± 0.96 1.3 ± 0.01 bcd 2.12 ± 0.15 d 1.04 ± 0.07 100
Within a column, means with different superscripts are significantly different (p<0.05).
Table 3: Hematological parameters of rainbow rout fingerling fed 8 weeks with diets containing
different levels of vitamin C and iron.
WBC Hematocrit Hemoglobi MCH MCHC
Treatments RBC (×106) MCV (fl)
(×103) (%) n (gr/dl) (pg) (gr/dl)
Control 101 ± 21 67.6 ± 15 36 ± 2.6 e 6.3 ± 0.49 299 ± 21.1 bcd 52.6 ± 6.3 17.5 ± 1.6
T1 97 ± 16.2 69.6 ± 8.9 33.3 ± 2.8 cde 6.5 ± 0.42 289 ± 38.2 bcd 56.4 ± 7.1 19.5 ± 0.2
cd d
T2 102 ± 21.5 69.3 ± 14 31.6 ± 1.1 5.7 ± 1.2 303.8 ± 18.7 59.1 ± 6.2 17.9 ± 2.2
T3 101 ± 10.3 64 ± 15 28.3 ± 1.5 ab 5.9 ± 0.13 316 ± 49.5 cd 59.5 ± 10.3 18.8 ± 0.36
cd ab
T4 103 ± 12.7 64 ± 16.2 32 ± 1 5.9 ± 0.81 244 ± 11.5 51.1 ± 6.7 20.9 ± 2.01
T5 101 ± 21.7 64.6 ± 15.5 30.6 ± 1.1 bcd 6.1 ± 0.11 300 ± 25.7 bcd 56.1 ± 8.7 18.6 ± 1.8
abc abc
T6 100 ± 13.9 61 ± 12.2 29 ± 4 5.8 ± 0.49 273 ± 10.2 54.2 ± 0.09 19.9 ± 0.76
a bcd
T7 102 ± 16 62.6 ± 9.2 28 ± 1 5.7 ± 0.04 298 ± 51.6 61.1 ± 15.8 20.2 ± 2.02
T8 103 ± 10.3 66 ± 7.5 33.3 ± 2.3 cde 6.2 ± 1.4 232 ± 20.2 a 47.9 ± 2.4 20.6 ± 0.75
e ab
T9 102 ± 17.3 64 ± 13.7 36 ± 2.8 6.3 ± 0.49 254 ± 14.1 46.9 ± 9.1 18.6 ± 4.7
Within a column, means with different superscripts are significantly different (p<0.05).
892 Adel and Khara, The effects of different dietary vitamin C and iron levels on the growth,…
Iranian Journal of Fisheries Sciences 15(2) 2016 892
Control 99 ± 1 1±1
T1 99.5 ± 0.58 0.67 ± 0.57
T2 98 ± 1.03 1±1
T8 99 ± 1 1±1
T9 99.6 ± 0.58 0.33 ± 0.58
Within a column, means with different superscripts are significantly different (p < 0.05).
160 f
140
IgM concentration (mg ml-1)
e e
120
100 d
d
80 d
c
60 bc
ab
40
20 a
0
50
45 f
Figure 2: Lysozyme activity of rainbow rout fingerling fed 8 weeks with diets
containing different levels of vitamin C and iron.
Studies with common carp (Sakamoto (2009) also have reported that addition
and Yone, 1978a), red sea bream of iron to the basal diet significantly
(Sakamoto and Yone, 1978b), Atlantic affected the Hct in juvenile gibel carp,
salmon (Andersen et al., 1996), channel C. auratus gibelio. Vitamin C is a
catfish (Barros et al., 2002) and powerful antioxidant protecting against
juvenile gibel carp, Carassius auratus oxidative damage to various tissues of
gibelio (Pan et al., 2009) did not fish including red blood cells (Sahoo
indicate an adverse effect of iron and Mukherjee, 2003). Lower red blood
deficiency on the growth performance. cells were regarded as the sun of
However, Shiau and Su, (2002) anemia which has been reported in most
reported that weight gain of Nile tilapia studies with fish where vitamin C
increased with dietary iron while deficiency was observed (NRC, 2011).
significantly decreasing at iron levels. Moreover, Adham et al. (2000)
These discrepancies might be due to demonstrated that feeds with
differences in individual size, insufficient vitamin C cause anemia,
development stage, and cultivation characterized by a decrease in the
environment. There were significant hemoglobin, reduction in the number of
differences in Hct and MCV values erythrocytes and hematocrit. Many
among our treatments. Similar results researchers have shown that ascorbic
were observed in pirarucu (Arapaima acid is involved in the metabolism of
gas) and cobia juvenile’s R. canadum iron in fish and a deficiency of ascorbic
(Zhou et al., 2012) fed with diet not acid has been observed to cause a
supplemented with vitamin C. In reduction in serum iron levels and a
agreement with our results Pan et al., redistribution of tissue iron stores in
Iranian Journal of Fisheries Sciences 15(2) 2016 894
rainbow trout (Hilton et al., 1989) and a examine the effect of dietary vitamins
reduction in both hemoglobin and and minerals on juvenile rainbow trout
hematocrit levels in catfish, trout and growth performance and
snakehead fish (Lim and Lovell, 1978; haematological parameters.
Agrawal and Mahajan, 1980). There are
also some as that may indicate the Acknowledgements
effect of iron on ascorbic acid "Financial support by Lahijan Branch,
metabolism in trout. Despite the Islamic Azad University Grant No.
interaction of ascorbic acid and iron 17.20.5.3511 is gratefully
metabolism in fish, it is interesting to acknowledged. "
note that increasing the levels of
ascorbic acid do not appear to affect the References
absorption of dietary iron in fish (Lanno Adham, K.G., Hashem, H.O. and
et al., 1985). The innate immune system Abu-Shabana, M.B., 2000. Vitamin
of fish is regarded to be the first line of C deficiency in the catfish Clarias
defense against pathogens and is more gariepinus. Aquaculture Nutrition,
important for fish as compared with 6(2), 129-139.
mammals. Lysozyme is released by Agrawal N.K. and Mahajan, C.L.,
leukocytes and plays a crucial role in 1980. Hematological changes due to
antimicroorganism activity. In the vitamin C deficiency in
present study, fish fed with different Channapunctut Block. Journal of
dietary vitamin C and iron levels had Nutrition, 110, 2172-2181.
lower lysozyme activity than fish fed Ai, Q.H., Mai, K.S., Zhang, C.X., Xu,
with the basal diets. Previous studies W., Duan, Q.Y. and Tan, B.P.,
indicated that the lysozyme activity 2004. Effects of dietary vitamin C on
have been found to be positively growth and immune response of
correlated with supplementation of Japanese seabass, Lateolabrax
ascorbic acid (Roberts et al., 1995; japonicus. Aquaculture, 242, 489-
Ortuno et al., 1999; Anbarasu and 500.
Chandran, 2001; Ai et al., 2004; Ai et Ai, Q.H., Mai, K.S., Tan, B.P., Xu,
al., 2006). In the present study, rainbow W., Zhang, W.B. and Ma, H.M.,
trout fed the basal diet had higher 2006. Effects of dietary vitamin C on
concentration than fish fed the other survival, growth, and immunity of
diets. Contrary to our results, Zhou et large yellow croaker, Pseudosciaena
al., (2012) showed that cobia, R. crocea. Aquaculture, 261, 327-36.
canadum fed with diet not Anbarasu, K. and Chandran, M.R.,
supplemented with vitamin C had lower 2001. Effect of ascorbic acid on the
concentration than fish fed the other immune response of the catfish,
diets. In conclusion, more studies using Mystus gulio (Hamilton), to different
basal diets containing different vitamins bacterins of Aeromonas hydrophila.
and minerals should be carried out to
IranianofJournal
895 Adel and Khara, The effects of Fisheries
different Sciences
dietary vitamin 15(2)
C and 2016
iron levels on the growth,… 895
Fish and Shellfish Immunology, 11, Halver J.E., 2002. The vitamins. In:
347-55. Halver, J.E., Hardy R.W., (Eds),
Andersen, F., Maage, A. and Fish Nutrition. Academic press,
Julshamn K., 1996. An estimation NewYork, 62-141.
of dietary iron requirement of Klontz, G.W., 1994. Fish Hematology.
Atlantic salmon, Salmo salar L., In: Techniques in Fish Immunology,
parr. Aquaculture Nutrition, 2, 41– Stolen, J. S.; Flecher, T. C.; Rowely,
47. A. F.; Zelikoff, T. C.; Kaattari S. L.
Azad, I.S., J.S. Dayal, M. Poornima and Smith S. A. (Eds.). Vol. 2, SOS
and S.A. Ali., 2007. Supra dietary Publications, USA. pp. 121-132.
levels of vitamins C and E enhance Haney, D.C., Hursh, D.A., Mix, M.C.
antibody production and immune and Winton, J.R., 1992.
memory in juvenile milkfish Chanos Physiological and haematological
chanos (Forsskal) to formalin-killed changes in chum salmon artificially
Vibrio vulnificus. Fish and Shellfish infected with erythrocytic necrosis
Immunolog, 23, 154-163. virus. Journal of Aquatic Animal
Barros, M.M., Lim, C. and Klesius Health, 4, 48-57.
P.K., 2002. Effect of soybean meal Hardie, L.J., Fletcher, T.C. and
replacement cottonseed meal and Secombes, C.J., 1990. The effect of
iron supplementation growth, vitamin E on the immune response
immune response and resistance of of the Atlantic salmon (Salmo salar
channel catfish (Ictalurus puctatus) L.). Aquaculture, 87, 1–13.
to Edwardsiella ictaluri challenge. Hilton, J.W., 1989. The interaction of
Aquaculture, 207, 263–279. vitamins, minerals and diet
Desjardins, L.M., 1985. The effect of composition in the diet of fish.
iron supplementation on diet Aquaculture, 79, 223-244.
rancidity and on the growth and Huang, C.H., Chang, R.J., Huang,
physiological response of rainbow S.L. and Chen, W., 2003. Dietary
trout. MSc thesis, University of vitamin E supplementation affects
Guelph, Ontario, Canada. tissue lipid peroxidation of hybrid
Durve, V.S. and Lovell R.T., 1982. tilapia, Oreochromis niloticus × O.
Vitamin C and disease resistance in aureus. Comparative Biochemistry
channel catfish (Ictalurus punctatus). and Physiology, 134, 265-70.
Canadian Journal of Fisheries and Jain NC.,1993. Essentials of Veterinary
Aquatic Science, 39, 948-951. Hematology, Lea and Feber,
Gouillou-Coustans, M.F., Bergot, P. Philadelphia, 417P.
and Kaushik, S.J., 1998. Dietary Lall, S.P, Oliver, G., Weerakoon,
ascorbic acid needs of common carp D.E.M., Hines, J.A., 1989. The
(Cyprinus carpio) larvae. effect of vitamin C deficiency and
Aquaculture, 161, 453-461. excess on immune response in
Atlantic salmon Salmo salar L. In:
Iranian Journal of Fisheries Sciences 15(2) 2016 896