The Effect of Replacement of Fish Oil (FO) By Canola Oil (CO) On Blood Serum Enzymes of the Rainbow Trout (Oncorhynchusmykiss)

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1 5, TextRod Puliction ISSN: 9-7 Journl of Applied Environmentl nd Biologicl Sciences The Effect of Replcement of Fish Oil (FO) By Cnol Oil (CO) On Blood Serum Enzymes of the Rinow Trout Elnz Nseri Khlkhli, Afshin Ghelichi *, Srh Jorjni Deprtment of Fisheries, Azdshhr Brnch, Islmic Azd University, Azdshhr, Irn, PO Box: Received: Mrch, 5 Accepted: July 7,5 ABSTRACT Ftty cids hve severl effects on immune responses. The present study ws performed in order to determine the effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum enzymes, i.e. Asprtte minotrnsferse (AST), Alnine minotrnsferse (ALT), Lctte dehydrogense (LDH), nd Alkline phosphtse (ALP), of the rinow trout.four experimentl diets were formulted with prtil or complete (, %, 67%, nd %) replcement of FOy CO.Fish were fed t % of their ody weight three times dy (t 8,, nd 8) for 5 dys.averge temperture, dissolved oxygen, nd ph were C, 7.5 mg.lit -, nd 7., respectively. Mesurement of ALT, AST, LDH, nd ALP ws performed y iochemistry nlyzer (Eppendorf, Germny) nd lortory kits. Sttisticl nlysis ws performed y one-wy ANOVA nd Duncn s multiple-rnge test (p<.5).the results derived from the present study reveled tht the fish fed y % fish oil hd the highest level of AST, ALT, LDH, nd ALP in their lood serum nd it hd significnt difference with other tretments (p<5). It is concluded tht oils used in fish feeding diets might hve profound influence on lood serum enzymes. KEYWORDS:Rinow trout, cnol oil, fish oil, serum enzymes. - INTRODUCTION Cytology is one of the most importnt rnches of clinicl studies which cn hve high level of contriution in clinicl dignoses. Mjor focus of cytologicl studies ws on red lood cells wheres other fctors such s white lood cells, proteins, nd ions re studied in current studies (Kzemi et l., ). Extrcellulr fluid, which covers ll cells of living creture, plys importnt roles in physicochemicl stility of intrcellulr sttus, providing energy sources for metolism, removing metolic wstes, nd molecule shifts, mong others (Ntochin et l., 997). Cells contin enzymes which ctlyze ll iochemicl rections in cells. Enzymes enter interstitil fluids nd the, they move to lood serum nd cererospinl fluid. Therefore, mesuring rte nd ctivity of enzymes in iologic fluids cn provide vlule informtion on functions of orgns nd tissues. Enzyme relese into lood circultion might e cused y severl resons such s cellulr necrosis nd noxi resulting in disintegrtion of cell memrne. So, chnges in orgns cn e detected y mesuring enzymes in lood serum. Interntionl mesurement unit for enzymes is Interntionl Unit per liter (IU.lit - ) (Moji, 99). Asprtte minotrnsferse (AST) exists in mitochondri nd plsm nd it is thus known s nonspecific enzyme. However, long with other enzymes, it cn help dignose liver nd muscle disorders. It is lso found in kidney tissue nd therefore, its increse cn not only e indictive of liver disorders ut lso kidney or even hert disorders. For instnce, it increses in nephroclcinosis in slmonide (Stoskopf, 99). Amount of AST does not chnge in Infectious Slmon Anemi (ISA) while it doules dy fter the infection in cyprinids. It is lso reveled tht trnsminses (e.g. AST nd ALT) rise in response to stress.alnine minotrnsferse (ALT)is found in plsm nd some cells. The mins source of this enzyme is liver nd its ctivity in lood serum rises considerly in cute heptic diseses leding to memrne dmge nd cell necrosis. ALT exists in fish lood serum nd it is undnt in fish liver. It rises considerly in cute liver necrotic disorder ut decreses in severe infection with Aeromons; it remin unchnged in noninfectious slmon nemi.lctte dehydrogense (LDH) is undntly found in ll tissues lthough it is 5 times more in red lood cells thn in plsm. It is nonspecific enzyme in liver, muscles, kidney, nd hert. It lso exists in fish with similr mechnism found in other nimls. Unlike AST, ALT, nd ALP, the mount of this enzyme rises in slmon nemi. Alkline phosphtse (ALP) is one of the first enzymes whose clinicl importnce hs een recognized. Its mount rises in liver nd one disorders. It lso increses in heptic firosis, nd heptic lipidosis. Use of immune stimultors to increse immune system ctivity hs gined gret del of ttention in quculture systems in order to enhnce resistnce ginst diseses in cultured fish. Severl immune stimultors *Corresponding Author: Afshin Ghelichi, Deprtment of Fisheries, Azdshhr Brnch, Islmic Azd University, Azdshhr, Irn, PO Box:. E-mil: Afshin.ghelichi@yhoo.com 55

2 Nseri Khlkhli et l.,5 hve een dopted in vccines nd foods to enhnce specific nd nonspecific immune responses, respectively. It hs een shown tht they hve importnt roles in protecting fish ginst disese (Kumri et l., ). Immune stimultors cn e prescried in foods for long time nd this cn e enefit over vccines. In ddition, some vccines cnnot e prescried for mny kinds of fish. The present study ws formulted in order to evlute the effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum enzymes of the rinow trout. - MATERIALS AND METHODS This study ws performed in Eductionl nd Reserch Center for Aquculture in Islmic Azd University, Azdshrh Brnch, Azdshhr, Irn. Fish were cultured in 5-lit fierglss tnks (5 5 cm). Wter ws stocked from deep well. The tnks were filled with wter until 5 cm. ech tnk ws equipped with cylindricl tues to close ottom hole of the tnks. Wter ws dischrged through the emedded holes t the end of cylindricl tues. Fish were purchsed from cold-wter quculture centers in Firoozkuh City. Three hundred fish (6.5±.6 g; 7±6. cm long) were selected nd trnsferred to the tnks. Twenty five fish were plced in ech tnk. In order to determine the effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum enzymes, four tretments (, %, 67%, nd % of CO insted of FO) in two itertions were considered. Fish feed (crude protein: 5., crude oil: 5.) ws purchsed from Mzndrn Cttle nd Fish Nutrition Co., Sri, Irn. Then, 5 g cnol nd/or fish oil ws dded to 85 g crude feed ccording to the tretments. The oil ws spryed on dry mtter nd then, the feed ws let to dry so tht the oil is sored y the feed. Fish were fed t % of their ody weight three times dy (t 8,, nd 8) for 5 dys. Tnk wter ws reduced to hlf while feeding to determine fish ppernce. Wter nd environmentl sttus, i.e. temperture ( C) nd dissolved oxygen (mg.lit - ), were recorded dily to mke sure environmentl sttus is similr for ll the tretments. Averge temperture, dissolved oxygen, nd ph were C, 7.5 mg.lit -, nd 7., respectively. At the end of the course, fish from e ch tnk ( fish from ech itertion) were selected. The fish were nesthetized y clove concentrte nd lood smples were tken from cudl vein. From ech smple, cc ws poured in tues without heprin. For serum seprtion, lood smples were centrifuged t rpm for 5 min. the serum were kept in freezer until next experiments. Mesurement of ALT, AST, LDH, nd ALP ws performed y iochemistry nlyzer (Eppendorf, Germny) nd lortory kits. Normlity test ws performed y Shpiro-Wilk test. Sttisticl nlysis ws performed y one-wy ANOVA nd Duncn s multiple-rnge test (p<.5) in SPSS (version ) nd grphs were drwn in Excel (Microsoft Office, 7). - RESULTS -- The effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum AST of the rinow trout The results showed tht the fish fed y % fish oil hd the highest level of AST in their lood serum nd it hd significnt difference with other tretments (p<5)(fig. ) tretment tretment Figure :The effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum AST of the rinow trout tretment tretment 56

3 -- The effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum ALP of the rinow trout The results reveled tht the fish fed y % fish oil hd the highest level of ALP in their lood serum nd it hd significnt difference with other tretments (p<5) (Fig. ) tretment tretment Figure : The effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum ALP of the rinow trout -- The effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum ALT of the rinow trout According to the results, the fish fed y % fish oil hd the highest level of ALT in their lood serum nd it hd significnt difference with other tretments (p<5) (Fig. ). tretment tretment 8 6 tretment tretment tretment tretment Figure : The effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum ALT of the rinow trout -- The effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum LDH of the rinow trout The results showed tht the fish fed y % fish oil hd the highest level of LDH in their lood serum nd it hd significnt difference with other tretments (p<5) (Fig. ). 57

4 Nseri Khlkhli et l.,5 6 8 tretment c tretment Figure : The effect of replcement of fish oil (FO) y cnol oil (CO) on lood serum LDH of the rinow trout - DISCUSSION AND CONCLUSION Effect of oil in feed hs not een studied extensively. Recent studies hve confirmed the effect of PUFA on fish immune system. The source of ftty cids entering plsm memrne is known s the oil in feed. Therefore, composition of ftty cid of cell memrne denotes oil composition of feed (Clmp et l., 997). Also, there is close correltion etween ftty cid composition in feed nd ftty cid composition in tissue (Srgent et l., ). Vritions in n-/n-6 in feed cn ffect immune cell structure (Thompson et l., 995; Bell et l., 99). They re energy source nd hve role in cell memrne. Mnipultion of feed cids cn influence on immune system y chnging structure of cell memrne (Tcon, ). The results otined from the present study showed tht AST, ALP, ALT, nd LDH in the fish fed with diet contining % fish oil were significntly higher thn those in other tretment. AST, ALP, nd ALT re nonplsm specific enzymes which re found not only in lood plsm ut lso in liver, hert, gills, kidney, muscles, nd other orgns. They cn lso provide vlule informtion out functions nd disorders in the orgns. The mount of AST, ALP, nd ALT is used s n indictor of liver function nd the enzymes re considered importnt to evlute helth sttus in fish (Rcicot et l., 975). LDH is often used to evlute heptic dmges (Yilmz et l., 6). Improper mngement of nutrition might effect on ALT ctivity (Cech et l., ). Activity of serum ALT nd AST vries with species. Increse in plsm AST nd ALT might e relted to stress condition, cell dmges in liver, nd cell degrdtion in liver, hert, or muscles cused y hevy metls (Yokoym et l., ). Menoyo et l. () found tht lipid oxidtion in rem liver fed y % nd % soyen oil ws lower thn tht in the fish fed y fish oil nd linseed oil. This is consistent with the results otined from the present study. In theory, the diets with high level of unsturted ftty cids re prone to oxidtion resulting in higher level of lipid peroxidtion in mitochondri nd heptic cell dmge (AedinKenri et l., ). Rehulk nd MinØik (7) stted tht increse in AST ctivity is sing of serious liver dmge vi AST relese in mitochondri. Infection rises ALT nd LDH. Skomoto et l. () showed tht chnge in enzyme mount cn e due to different lood smpling techniques, nlysis methods, ge, diet, nd hitt. LDH mount in the fish fed y the diet with % fish oil ws significntly higher thn tht in other tretments. High sturtion level in the diet might cuse shrinkge in red lood cells (Mourente et l., 6). Khjeh nd Peyghn (7) nlyzed some iochemicl fctors of lood serum of rinow trout cultured in erth ponds nd reported serum ALP, AST, ALT, nd LDH to e 7±, 7±5, 9±, nd 579±5 IU.lit -. The results cquired from the present study out ALP nd ALT were lower thn those in the study of Khjeh nd Peyghn (7). LDH in the present study in ll the tretments ws higher thn tht reported y Khjeh nd Peyghn (7). As feeding ws performed y hnd here nd the food ws kept t mient sitution, lipid oxidtion, especilly in fish oil, might hve dverse effect on red lood cells nd LDH relese resulting in higher level of this enzyme in plsm. However, morphologicl studies on red lood cell is required to prove this. In conclusion, oils used in fish feeding diets might hve profound influence on lood serum enzymes. However, hygienic issues should e considered during oil preservtion, especilly fish oil, ecuse ftty cid oxidtion not only rouses immune system response ut lso wekens immune system nd might hve considerle influence on lood serum enzymes. tretment tretment 58

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