Effects of super supplements on growth and some physiological indices in farmed juvenile Siberian sturgeon (Acipenser baerii)

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1 Interntionl Journl of Fisheries nd Aquti Studies 2016; 4(6): ISSN: (ICV-Polnd) Impt Vlue: 5.62 (GIF) Impt Ftor: IJFAS 2016; 4(6): IJFAS Reeived: Aepted: Rhin Alsdt Hsehemi M.S. Grduted, Islmi Azd University, Rsht Brnh, Irn Mhmoud Bhmni Fulty Memer, Interntionl Sturgeon Reserh Institute, Agriulturl reserh, edution & extension orgniztion (AREEO), Tehrn, Irn Ayou Yousefi Jourdehi Fulty Memer, Interntionl Sturgeon Reserh Institute, Agriulturl reserh, edution & extension orgniztion (AREEO), Rsht, Irn Correspondene Rhin Alsdt Hsehemi M.S. Grduted, Islmi Azd University, Rsht Brnh, Irn Effets of super supplements on growth nd some physiologil indies in frmed juvenile Sierin sturgeon (Aipenser erii) Rhin Alsdt Hsehemi, Mhmoud Bhmni nd Ayou Yousefi Jourdehi Astrt This study ws rried out with the im of determining the effets of dietry super supplements on some physiologil nd growth indies in juvenile Sierin sturgeon (Aipenser erii). A totl of 180 fish fter two weeks of dpttion, were rndomly distriuted in 15 fierglss tnks (2000 liters). 12 fish were introdued to eh tnk. supplements t 2.5%, 5% nd 10% were dded to the diet nd experiments were performed in triplite. Smples were olleted for the mesurement of growth, lood, immune nd osmoti indies during the rering period. The results showed tht the verge finl weight, finl iomss, ody weight inresing (BWI), speifi growth rte (SGR), ondition ftor (CF) nd dily growth rte (ADG) showed signifint differene etween ontrol group with treted ones (P<0.05). Aording to the results, the use of super supplement improved growth nd immune indies, prtiulrly in the tretment ontining 5% super supplement nd dding it is reommended to the diet of A. erii. Keywords: supplements, hemtologil prmeters, immune system, growth, sierin sturgeon 1. Introdution During the lst ten yers, ntiiotis re ommonly eing used for the mngement of quti niml diseses, s well s for inrese in growth nd improvement of feed onversion rtio [11]. Antiiotis ffeted the nturl flor in fish nd inrese their resistne to them. Moreover, ntiiotis re hemil mtters tht negtively ffet on the environment [22]. It is lso possile to ntiiotis residul umulted in quti nimls [13]. Detils of history of orgni quulture nd its stndrds were presented for the first time in the world [9]. Menwhile, the first model of orgni frming in 1994 ws reorded in Austrli nd Germny for rp. It is ovious tht onsidering to the orgni quulture in the Asin mrkets is developed more thn other prts of the world in reent yers whih its reson is the high per pit onsumption of sefood in Asi nd its historil kground. The ommeril vlue of orgni produts is 1.7 to 4 times ompred to the onventionl produts [27]. So, if Irnin produers re le to produe nd export orgni produts, the vlue of griulturl exports will e 2 to 4 times. With the prodution nd export of orgni produts the dded vlue of produts will inrese. Orgni quulture inludes omponents tht ensure tht quulture tivities re omptile with the environment nd orgnism is in helth nd wellness onditions, tody orgni produts in the quti nimls industry y super supplements hs found speil ple mong onsumers, the use of hemils, hormones, ntiiotis nd rooders tht re genetilly modified, is inomptile with the priniples of orgni quulture [27]. Supplements re nutrients tht onsisting of ft-solule vitmins (Vit. A, D 3, E, K), essentil minerls: lium, phosphorus, iron, opper, nesium, mngnese, olt, zin, iodine, selenium, hlorine, sodium, potssium, growth promoters: etfin, holine, rnitine, oted vitmin C, oted miro-ionized proteins (these re indigestile proteins), essentil mino ids: lysine, methionine, ysteine, threonine nd essentil ftty ids: linoleni, rhidoni nd linoleni tht re used in fish nutrition. One of the most importnt nd relile inditors of helth sttus nd physiology of fish is mesuring of hemtologil, immunity nd osmoti prmeters tht ffeted y nutrition, environmentl ftors, nd et. [12]. Sierin sturgeon n e esily dpted to ultured onditions, is resistnt to environmentl hnges, hs high growth ~ 400 ~

2 Interntionl Journl of Fisheries nd Aquti Studies rte nd esily use ville food soures. Beuse of low mturity ge nd rpid vir rehing, this speies is used in mny studies nd is one of the importnt speies tht is used in the quulture industry. Sierin sturgeon is sturgeon speies tht hs high growth rte nd is very suitle for quulture, ut little reserh hs een done on its nutritionl requirements. In this study, we hve tried to evlute the impt of super supplements on physiology nd growth prmeters of Sierin sturgeon. The results of present study leds to ommeril diet nd improves the ulture onditions [31]. 2. Mterils nd Methods In this study, 180 Aipenser erii (with men weight ± gr nd men length ± 1.18 m) were provided from the Interntionl Sturgeon Reserh Institute. After two weeks dpttion, fish were rndomly introdued to 15 fierglss tnks with volume of 2000 litres. Fish were fed with Skretting diet sed on tle1: Tle 1: Skretting diet nlysis sed on presented speifitions y Skretting Compny Ingredients Perent (%) Crude Protein 46 Crude lipid 17 Crohydrte 18 Ash 8.5 Fier 2.5 Moisture 10 In ddition, super supplement from ATA Compny with tle 2-2 nlysis nd Oxy-tetryline ntiioti 20% from Ars Brn Compny were used. This study ws onduted in five tretments with triplites, inluding ontrol without ntiioti, ontrol with ntiioti nd super supplement t 2.5%, 5%, nd 10% for 90 dys nd three times per dy [7, 42]. Tle 2: Anlysis of rw mterils of supplement (ATA Compny) Vlue unit Mg Mg Composition Copper (Cu) Ferrous (Fe) Zin (Zn) Selenium (Se) Iodine (I) Colt (Co) Ntrium (N) Chlor (Cl) Phosphorus (P) Clium (C) Anti-Oxidnt Vlue Unit Mg 1400 Kl/kg 2000 Mg Mg Mg 1400 Kl/kg 2000 Mg Mngnese (Mn) Mgnesium (Mg) Composition L-Crnitin Betein Choline Multi enzyme Net Protein Arginine Cystine Methionine Lysine Gross Energy Lionolei id Vlue unit IU IU 0 Vit.A Vit.D3 Vit.E Vit.K Vit.B1 Vit.B2 Vit.B3 Vit.B5 Vit.B6 Vit.B9 Vit.B12 Vit.H2 Vit.Srey C Evlution of growth trend ws lulted sed on the ville referenes of mthemtil equtions [8]. (ADG) (g/fish/dy) (%) = [w t w i /w i T] 100 Averge dily growth Perent ody weight inrese (PBWI) (%) = [w t w i / w i] 100 Speifi growth rte (SGR) (dy) = [LnWt - LnW i / T] 100 Feed Conversion Rtio = Feed intke (g)/ w f w i During experiment the physil nd hemil prmeters of wter (temperture, ph, dissolved oxygen nd totl hrdness of wter) were mesured nd reorded on dily sis y multimeter (WTW Germny) (tle 3). Tle 3: Physil nd hemil prmeters of wter during 90 dys. Prmeters Temperture ( C) ph Dissolved oxygen (/l) Wter hrdness ( /l) CCO3 Quntity 15.73± ± ± ±2.44 Blood smples were olleted in the middle (45 dys) nd the end (90 dys) of experiment period using 2 ml syringe under the udl fin. Plsm seprted from lood ells y using entrifuge (Lofuge, 200, mnuftured y Hereus Septeh Germny) t 3000 rpm for 10 minutes [29]. Mesurement of lood, immune nd osmoti prmeters ws onduted in physiology nd iohemistry Deprtment of interntionl Sturgeon Reserh Institute, Rsht. Counting of white lood ells (WBC) nd red lood ells (RBC) ws done y hemoytometer (Neuuer) [7, 30]. Blood smers were prepred for differentil ount of white lood ells. Smers were fixed with methnol nd stined with Giems nd ells were ounted under light mirosope [26]. Hemtorit ws mesured y hemtorit method [16]. The level of hemogloin ws mesured y olorimetri method using spetrophotometer t wvelength of 560 nm [6, 16]. Hemtologil prmeters inluding men orpusulr volume (MCV), men orpusulr hemogloin (MCH) nd men orpusulr hemogloin onentrtion (MCHC), were otined from the following equtions [16]. MCV (fl) = (Hemtorit / RBC) 10 MCH (pg) = (Hemogloin / RBC) 10 MCHC (g /dl) = (Hemogloin / Hemtorit) 100 Mesurement of immunogloulin M nd lysozyme level ws performed using the Elis reder (Awreness, USA, model Stt fx) nd turidimetri method. Prs Azmoon test kit nd turidity method used for the mesurement of omplement C 3 nd C 4. Automti osmometer (Nr Type-13, mnuftured y Roeling, Germny) ws used for mesuring serum osmollity. Sodium nd potssium ions were mesured y Flme Photometer (C405) of Fster Eletri nd mesurement of lium ion ws done y Prs Azmoon kit nd Autonlyzer (RA1000 model). Experimentl dt were exmined y normlity test. To determine signifint differenes one-wy nlysis of vrine (Anov) ws used nd for non-norml dt Kruskl - Wllis ws used. One-wy nlysis of vrine nd Kruskl ~ 401 ~

3 Interntionl Journl of Fisheries nd Aquti Studies Wllis were used to determine signifint differenes in lood prmeters were norml. Sttistil nlysis nd drwing grphs ws done through softwre SPSS 16 nd Exel 2010 [3]. 3. Results 3.1. Growth prmeters The men of initil weight, iomss, length, finl length nd FCR showed no signifint differene etween ontrol nd other tretments (P>0.05). The men of finl weight t onentrtion 5% nd 10% ws signifintly higher thn the others (P<0.05). The men finl iomss, men ody weight inresing (BWI%), speifi growth rte (SGR), dily growth rte (DGR) nd ondition ftor (CF) in super supplement 5% nd ontrol tretment without ntiioti ws signifintly more thn other tretments (P<0.05) (tle 4). Tle 4: Growth prmeters of vrious tretments in Sierin sturgeon during experimentl period. Growth prmeters Control Without ntiioti ntiioti supplement 2.5% supplement 5 % supplement 10 % Initil weight (g) ± ± ± ± ±4.17 Finl weight (g) ± ± ± ± ±20.88 Initil Biomss (g) ± ± ± ± ±50.03 Finl Biomss (g) ± ± ± ± ± Initil length(m) ± ± ± ± ±0.26 Finl length (m) ± ± ± ± ±0.21 BWI (%) ± ± ± ± ±1.86 FCR 1.71 ± ± ± ± ±0.08 SGR (dy) 0.73 ± ± ± ± ±0.01 GR 7.78 ± ± ± ± ±0.03 CF 0.71 ± ± ± ± ± Hemtologil indies There ws signifint differene in the totl numer of white lood ells in the ontrol group with other tretments exept for super supplement 5% (P<0.05). In the first phse of ulture period, white lood ell differentil ount showed no signifint differenes etween numer of lymphoytes, monoytes nd eosinophils in ontrol ompred to the other tretments (P>0.05). There ws signifint differene etween the neutrophils in ontrol nd onentrtion 10% of super nutrient (P<0.05). There ws signifint differene in the numer of red lood ells etween the ontrol with other tretments exept for super supplement 5% (P<0.05). Hemtorit showed signifint differene etween ontrol nd other tretments (P<0.05). The highest hemtorit level ws oserved in the ontrol group inluding ntiioti (P<0.05). No signifint differenes were oserved etween the ontrol group with the other tretments in the mount of hemogloin level nd lood indies (indies (Men Corpusulr Volume, Men Corpusulr Hemogloin nd Men Corpusulr Hemogloin Conentrtion) (P>0.05) (tle 5). Tle 5: Results of hemtologil indies in the first ulture phse of Sierin sturgeon. Hemtologil indies WBC ( n/mm 3 ) Lymphoyte (%) Neutrophil (%) Eosinophil (%) Monoyte (%) RBC (n/mm 3 ) Ht (%) H (gr/dl) MCV (fl) MCH (pg) MCHC (%) Control Without ntiioti ntiioti supplement 2.5% supplement 5% supplement 10% 5700 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.66 2± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.00 Aording to tle 5, there is signifint differene etween the totl numers of white lood ells in the ontrol group with the other tretments exeption of super supplement 2.5% (P<0.05). White lood ell differentil ount showed tht there ws no signifint differene etween numer of lymphoytes, neutrophils, monoytes nd eosinophils in ontrol with other tretments (P>0.05). RBC showed no signifint differene in ontrol group nd the other tretments (P>0.05). Signifint differenes were oserved in the hemtorit level etween ontrol ompred with the other tretments nd the mximum level ws oserved in ontrol with ntiioti nd super supplement 2.5%, whih ws signifintly more thn the other tretments (P<0.05). No signifint differene ws oserved in the mount of hemogloin nd other lood indies (Men Corpusulr Volume (MCV), Men Corpusulr Hemogloin (MCH) nd Men Corpusulr Hemogloin Conentrtion (MCHC)) etween the ontrol with the other tretments (P>0.05) (tle 6). ~ 402 ~

4 Interntionl Journl of Fisheries nd Aquti Studies Tle 6: Results of lood indies of Sierin sturgeon in the finl ulture phse. Hemtologil indies WBC ( n/mm 3 ) Lymphoyte (%) Neutrophil (%) Eosinophil (%) Monoyte (%) RBC (n/mm 3 ) Ht (%) H (gr/dl) MCV (fl) MCH (pg) MCHC (%) out ntiioti ntiioti supplement 2.5% supplement 5% supplement 10% ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.33 As shown in tle 7, the differene in the mount of immunogloulin M ws not signifint in ontrol with the other tretments with the exeption of super supplement 5% tretment (P<0.05). C 3 showed signifint differene in ontrol with the other tretments (P<0.05), ut signifint differenes were not oserved in C 4 onentrtion etween tretments (P>0.05). Differene ws not signifint in the mount of serum eletrolytes nd osmollity of Sierin sturgeon in the first ulture phse etween ontrol group with ny of the tretments (P>0.05) (tle 7). Tle 7: Immune nd osmoti indies in Sierin sturgeon t the first ulture phse. Immune nd osmoti indies IgM (/dl) Lysozyme (μg/ml) C3 (μg/ml) C4 (μg/ml) C 2+ (μg/ml) N + (/dl) K+ (/dl) Osmolrity (mosml/l) out ntiioti ntiioti supplement 2.5% supplement 5 % supplement 10 % ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±9.68 d ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.09 3± ± ± ± ± ±0.88 There ws signifint differene in IgM level etween the tretment super supplement 5% with other tretments (P<0.05). At the finl phse of ulture, there re no signifint differenes in the mount of lysozyme nd C 3 etween ontrol group nd other tretments (P>0.05). There ws signifine differene in the mount of C 4 in ontrol with the other tretments (P<0.05). Serum eletrolytes nd osmollity hnges showed no signifint differene etween ontrol with the other tretments t the finl phse of ulture (P>0.05) (tle 8). Tle 8: Immune nd osmoti indies of Sierin sturgeon in the finl ulture phse. Immune nd osmoti indies out ntiioti ntiioti supplement 2.5% Supplement 5% supplement 10% IgM (/dl) 3.17 ± ± ± ± ±31 Lysozyme (μg/ml) 4.50 ± ± ± ± ±34 C3 (μg/ml) 3.17 ± ± ± ± ±40.33 C4 (μg/ml) 1.45 ± ± ± ± ±34 C 2+ (μg/ml) 4.91 ± ± ± ± ±8.93 N + (/dl) 1.20 ± ± ± ± ± K + (/dl) 0.15 ± ± ± ± ±2.23 Osmolrity (mosml/l) 1.52 ± ± ± ± ± Disussion In the present study, the most effet of super supplements on growth prmeters suh s men finl weight, finl iomss, inresed ody weight (BW%), speifi growth rte (SGR), dily growth rte (GR) nd ondition ftor (CF) ws oserved in tretment ontining super supplement 5%. In this study the durtion of feeding storge ws short. This lso helps to keep the food qulity whih is of the si stndrds in orgni fish prodution [27]. studied the impt of orgni ulture with 2.5 nd 5% super supplement on some growth prmeters in Common Crp (Cyprinus rpio) nd found tht tht verge finl weight, weight gin, ondition ftor, speifi growth rte nd verge dily growth showed signifint differenes [27] (P<0.05). ~ 403 ~ The results of experiments for evlution of different levels of etine nd methionine, s n ttrtnt, in the growth prmeters of juvenile elug showed tht dding supplementtion in the diet uses inresing in weight, speifi growth rte nd ondition ftor [36]. Reported tht simultneous use of vitmin C nd proioti; Billus sutilis in Ruho (Leo rohit) n signifintly inrese growth prmeters. Positive role of minerl supplements hs een proven in nutrition of quti nimls to hve norml nd helthy life; fish provide these minerls from food nd wter. In this study, n inrese in growth prmeters suh s BWI %, SGR, GR nd CF oserved in the tretment super supplement 5% ompred to other tretments, espeilly ontrol with ntiioti my e due to inresed food intke

5 Interntionl Journl of Fisheries nd Aquti Studies nd improved digestion proess euse of vrious nutrients, espeilly vitmins nd minerls in super supplement tht stimultes growth in fish, proly due to hnge in the omposition of nutrients in the tretment super supplements 5%. Vrious ftors suh s the omposition of nutrients in the diet, method of feeding, environmentl ftors suh s wter temperture nd dissolved oxygen, fish size [17], nd the omintion of ingredients [33] ffeted feed onversion rtio (FCR). Blood is one of the physiologil prmeters of quti nimls s fluid tissue nd is one of the most importnt iologil liquids nd its omposition is hnged under influene of vrious physiologil nd pthologil sttes [20, 32]. Blood prmeters re inditors for determining the helth sttus of quti nimls [37]. Bsed on the otined results in the first phse, the lrgest numer of white lood ells oserved in tretments inluding super supplements 2.5%, 10% nd ontrol with ntiioti. In the finl phse of ulture, the highest numer of white lood ells ws oserved in the ontrol with ntiioti nd super supplements 5% nd 10%. The numer of red lood ells n hve signifint effet on the lne of whole energy of the ody [34]. In study, the effet of super supplements on the lood nd immune indies of grss rp (Ctenophryngodon idell) ws investigted nd smpling rried out in four phses nd found tht the fish lood immunogloulin were signifintly different etween tretments nd ontrol in the seond nd third phses of smpling [25]. Vitmins in super supplements suh s vitmin C is one of nturl nti-stress ompounds in fish tht hs numerous roles [43], inluding the stimultion of the immune system omponent suh s lterntive omplement pthwy nd serum lysozyme tivity [39]. Studied the effet of different levels of vitmin supplement ontining vitmins C nd E on the immune prmeters of Cspin trout, nd reported n inrese in immunogloulin nd lysozyme tivity [35]. Investigted vrious doses of vitmin C on hyrid sturgeon (A. ruthenus & A. erii) nd onluded tht lysozyme tivity showed signifint differenes in fish were fed y different levels of vitmin C [18]. Therefore, vitmin C hs positive effet on lood serum lysozyme tivity in fish. The hnges in lysozyme s one of the most non-speifi immune prmeters is so speil tht will inrese prodution of the enzyme lysozyme. Inresing in the prodution of the enzyme lysozyme is due to the presene of vitmin C in vitmin supplement. In the first phse of ulture, there ws signifint differene etween the mount of lysozyme in the ontrol group ompred with super supplement 5 nd 10%. But t finl phse of ulture, signifint differenes were not oserved in the mount of lysozyme etween ontrols with other tretments. Level of C 3 hs signifint differene in the ontrol group with the exeption of super supplement 5%, ompred with other tretments. At the finl phse of ulture, signifint differenes were not oserved in the level of C 3 in the ontrol group nd other tretments. Level of C 4 in ontrol group with the other tretments showed no signifint differene, ut there ws signifint differene in the C 4 level in ontrol group ompred with other tretments ut in the finl phse of ulture. Complements re the most importnt defense ftors tht re out 35 solule proteins, whih ply key role in the innte nd dptive immunity [10]. The most importnt iologil funtions of the omplement system onsists of removing miroorgnisms through prtiiption nd oopertion in the proesses of phgoytosis, inflmmtory response, lering immune omplexes, indution nd enhnement of ntiody responses [15, 21]. Mny immunostimulnts inresed omplement tivity in the fish serum [24]. Chnges in the serum omplement to the protetion of non-speifi immune system in fish re very importnt nd high levels of C 3 nd C 4 indited the helth of the fish [41]. studied the simultneous use of vitmin C nd proioti Ltoillus rhmnosus on immune system of rinow trout (Onorhynhus mykiss) whih inresed the serum lysozyme tivity nd omplements (C 3 nd C 4) [38]. Osmoti indies influened y environmentl ftors, espeilly dissolved oxygen, slinity, temperture nd so on. Mny of the fish showed intertions in order to mintin its genertion y hnging the environmentl onditions [2]. Considering to the mentioned ftors, supplements not ffeted on serum osmolrity nd eletrolyte indies. Investigted the effet of feeding rtio on some iohemil prmeters of rp (Cyprinus rpio) fingerlings. The results of this study showed tht the frequeny nd perentge of feeding hd no effet on the level of serum lium in rp fingerlings [23]. In generl, it n e onluded from this study tht the use of super supplement in the diet improves the onsidered prmeters. Tretment super supplement 5% ompred to tretment with ntiioti showed etter growth nd immunity. Therefore, the use of super supplement in the diet s n dditive stimultes the growth nd immune system. It is reommended tht it should e used in omintion with diet for Sierin sturgeon. With regrds to the persistene of hemil sustnes, espeilly drugs nd ntiiotis in the odies of frmed fish, it seems tht it is essentil to e onsidered the prodution of orgni food produts due to the importne of the food helthy nd sfety in the soiety. 5. Referenes 1. Aikins F SGH. Effets of feeding high level of D- gluose on liver funtion in juvenile white sturgeon (Aipenser trnsmontnus). Fish Physiology Biohemil. 1993; 12: Akimov NV, Run GI. The ondition of the reprodutive System of the Sierin sturgeon, Aipenser reii, s ioinditor. Journl of Ihthyology. 1993; 33: Atmnlp M, Ynik T. 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