Influence of Aluminum Chloride and Ascorbic Acid on Performance, Digestibility, Caecal Microbial Activity and Biochemical Parameters of Rabbits

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1 Reserh Journl of Agriulture n Biologil Sienes (): 0-6, , INSInet Pulition Influene of Aluminum Chlorie n Asori Ai on Performne, Digestiility, Cel Miroil Ativity n Biohemil Prmeters of Rits S.M.A. Sllm, M.E.A. Nsser, M.S.H. Yousef, A.M. El-morsy n S.A.S. Mhmou, M.I. Yousef. Deprtment of Animl Proution, Fulty of Agriulture, Alexnri University, Egypt. 2 Centrl lortory for Foo n Fee, Agriulture Reserh enter, Egypt. Deprtment of Animl Proution, Fulty of Agriulture, Suez Cnl University, Egypt. 4 Deprtment of Environmentl Stuies, Institute of Grute Stuies n Reserh, Alexnri University, Alexnri, Egypt. Astrt: The effets of luminum hlorie (AlCl ), sori i (AA) n/or their omintion were investigte on the performne, igestiility, nitrogen lne, el miroil tivity n iohemil prmeters in mle New Zeln white rits. The experimentl rits were rnomly ivie into four groups (n=6/ group). The first group ws use s ontrol, the seon group ws use to stuy the effet of sori i, the thir group ws use to stuy the effet of luminum hlorie, while the fourth group ws given the omintion of sori i n luminum hlorie. The teste oses of luminum hlorie n sori i were given every other y for 6 weeks. The tretment with AlCl resulte in signifint (p<0.05) erese in oy weight, fee intke, rinking wter, nitrogen lne, igestiility oeffiients (DM, OM, CP, CF, EE n NFE ), TDN n DCP. Also, tretment h signifint (p<0.05) effets onthe tivities of sprtte minotrnsferse (AST), lnine minotrnsferse (ALT), lkline phosphtse (AlP), i phosphtse (AP) n the onentrtion of thiorituri i-retive sustnes (TBARS) plsm enzymes tivity. But, presene of AA lone or omintion with AlCl oul e inition of the improve liver funtion n protetive from the toxiity of luminum. Key wors: Rit, sori i, luminum hlorie, igestion, iohemil prmeters, INTRODUCTION utensils of pkging, foo n everge n [] luminum-ontining meitions. Aluminum (Al), the most unnt metl, omprising Minmi et [4] l. oserve n ge-epenent out 8% of the erth's rust, is foun in omintion umultion of luminum in the ort n ererl with oxygen, silion, fluorine n other elements in soil, [5] rteries. Bush et l. reporte inrese onentrtion of [] roks, lys n gems. Aluminum metl is use s luminum in serum, one, liver n spleen n slight struturl mteril in the onstrution, of utomotive n inrese in the rin. Aluminum hs een propose s n irrft inustries, in the proution of metl lloys n in environmentl ftor tht my ontriute to some the eletriity inustry in power lines, insulte les neuroegenertive iseses n ffets severl enzymes n wiring. Other uses of luminum metl inlue ooking n other iomoleules relevnt to Alzheimer s isese. utensils, eortions, fening, highwy signs, ns, foo Slts of luminum my in to DNA, RNA, inhiit suh [2] pkging, foil n entl rowns n entures. enzymes s hexokinse, i n lkline phosphtse, Aluminum is relese to the environment y oth nturl [6] phosphoiesterse n phosphooxyse. proesses n nthropogeni soures, 5,605,000 I of Chemil toxints my lter nutrients intke, luminum ws relese to the environment (ir, wter n igestion, sorption, trnsport, or funtion metolism. soil) from 264 lrge proessing filities. Of this totl, In generl, hemils toxints n ffet nutrition y out 0, 0.9, 69% were relese to the ir, wter n soil, ltertion of the nutrient ontents of fee. Toxints my respetively. erese fee intke y moultion of ppetite. In generl Humn exposure to lrge quntity of luminum in sense, ny sustne whih isturs metolism or nture n its mny uses is me through intke of mjor physiology suffiiently to use growth inhiition uses soures i.e. rinking wter, foo resiues, ooking erese fee intke. These sustnes lso inhiit ATP Corresponing Author: S.M.A. Sllm, Deprtment of Animl Proution, Fulty of Agriulture, Alexnri University, Egypt 0

2 Res. J. Agri. & Biol. Si., ():0-6, 2005 proution n therefore inhiit protein n tive trnsport, with finl result of erese oy weight n [7] fee intke. Reent stuies were rrie out to evlute the potentil role of ntioxint vitmins, suh s vitmin C, [8,9] vitmin E n $-rotene. Vitmin C (sori i) is n essentil mironutrient require for norml metoli funtioning of the oy. Mny iohemil, linil n epiemiologi stuies hve inite tht vitmin C my e of enefit in hroni iseses suh s riovsulr isese, ner n trt, proly through [0] ntioxint mehnisms. Vitmin C is oftor for severl enzymes involve in the iosynthesis of ollgen, rnitine n [,2] neurotrnsmitters. In ition, vitmin C is use s oftor for teholmine iosynthesis, in prtiulr the onversion of opmine to norepinephrine tlyze [] y opmine "-monooxygense. Also,Vitmin C is reuing the negtive effets of fltoxin B on [9] proution n reproution. Vitmin C prevents free ril mge in the lungs n my even help to protet the entrl nervous system from suh mge. It ts ginst the toxi, mutgeni n rinogeni effets of environmentl pollutnts y stimulting liver etoxifying [] enzymes. Therefore, this work ime of stuying the effet of luminum hlorie, vitmin C n their omintion on rits performne, igestion, miroil tivity, free rils n enzyme tivities in plsm. MATERIALS AND METHODS This reserh ws rrie out t Hormone Lortory, Deprtment of Environmentl Stuies, Institute of Grute Stuies n Reserh, Alexnri University. In this stuy luminum hlorie (AlCl ) n sori i (vitmin C, liqui supplement) were use. Aluminum hlorie ws purhse from Alrih Chemil Compny (Milwukee wis, USA) n vitmin C (20%) ws purhse from Neofrm, Itly (Vi Emili Km 8, No Longino, Fo, Itly). Twenty four mle New Zeln White rits (ge of 7 months n initil weight of 2920±42 g) were use. Animls were iniviully house in ges n weighe weekly throughout 6 weeks experimentl perio. The [4] hemil nlysis of the pellets showe t Tle. Rits were rnomly ivie into four equl groups (n = 6/group). The groups were ontrol, sori i (AA, 40 kgg oy weight), luminum hlorie (AlCl, 4 kgg oy weight, /25 LD 50 ) n AA plus AlCl. The LD50 of luminum hlorie when given orlly to [5] rits ws reporte to e 400 kgg BW. Tle : Approximte hemil nlysis of ommeril onentrte mixture (% s DM sis) Items % DM sis Orgni Mtter Ash 7.2 Crue Protein 5.77 Crue Fier.2 Ether Extrt.52 Nitrogen Free Extrt 62.6 Bloo smples were ollete from the er vein of ll nimls every week throughout the 6 weeks experimentl perio. Bloo smples were otine in the morning efore esses to fee n wter n ple immeitely on ie. Heprin ws use s ntiogulnt. Plsm ws otine y entrifugtion of smples t 860 g for 20 min n ws store t -20 C until use for nlysis. Store plsm smples were nlyze for Thiorituri Ai-retive Sustnes (TBARS) were mesure in [6] the plsm y Tppel n Zlkin. The tivities of plsm sprtte minotrnsferse (AST) n lnine minotrnsferse (ALT) were ssye y the metho of [7] Reitmn n Frnkel. While lkline phosphtse (ALP) n i phosphtse (ACP) ws etermine oring to [8] [9] Prinipto et l. n Moss, respeyively. The Glutthione S-trnsferse (GST) n Aetylholinesterse [20] (AhE) ws estimte y Hig et l. n Ellmn [2] et l., respetively. During the lst week of the experimentl perio, the rits of eh group, fe the sme iet, were iniviully house in metoli ges for igestiility n nitrogen lne etermintions [22] oring to F ekete. These nimls were kept for 7 ys s olletion perio. During this perio, fees n urine were quntittively ollete. Spot smples of fresh urine were store frozen t -0 C for the nlysis of totl N. Fees from eh niml ws poole over the 7- perio for etermine ry mtter n hemil nlysis. Cel ontents were remove immeitely fter slughter n store frozen t -0 C until nlysis. The omplete hemil omposition of fee, fee resiue, fees n ry el ontents were etermine oring to the [4] AOAC. Nitrogen ws etermine y miro-kjelhl metho. Totl Voltile Ftty Ais (VFA) in the el ontents were estimte y stem istilltion n [2] titrtion. Ammoni-N ws etermine in the wet [24] el smples y mgnesium oxie methos. Dt of the experiment for ll vriles were sujete to ANOVA s ompletely rnomize [25] esign oring to Sneeor n Cohrn. The signifint ifferenes etween tretment mens were teste y the Dunn, Lest Signifint [26] Rnge Test.

3 Tle 2: Effet of sori i (AA), luminum hlorie (AlCl ) n their omintion on live oy weight, ry mtter intke (DMI) n wter intke of mle rits (mens±se) Items Control AA AlCl AA+AlCl No. of nimls Initil oy weight (g) 2905± ± ± ±6.0 Finl oy weight (g) 400 ±.0 52 ± ± ±60.0 DMI (g G ) 7. ± ± ± ±02.5 Wter intke (ml G ) 84 ± ± ± ±0.8 Tle : Digestion oeffiients n nutritive vlues of mle rits trete with sori i (AA), luminum hlorie (AlCl ) n their omintion (mens±se) Items Control AA AlCl AA+AlCl Res. J. Agri. & Biol. Si., ():0-6, 2005 RESULTS AND DISCUSSION rinking wter ompre to the ontrol group (Tle ). Also, previous stuies showe tht sori i Boy weight, fee intke n rinking wter: The results [9] supplementtion stimulte weight gin in rits. of oy weight, fee intke n rinking wter (DW) re [] Yousef et l. foun tht vitmin C use signifint presente in Tle 2. Men vlues inite tht (p<0.05) inrese in FI, while the inrese in BWG ws tretment with AlCl use signifint (p<0.05) erese insignifint of rits, the enefiil effets of sori in Live Boy Weight (LBW), Fee Intke (FI) n rinking i whih ws note in the present stuy n e wter s ompre to ontrol. Asori i lone h ttriute to the ntioxint effets of this vitmin; it is signifint (p<0.05) inrese in LBW, FI n rinking svenge of oxygen free rils whih re toxi ywter. The omintion of AA with AlCl use inrese prouts of mny metoli proesses. [4-6] in the reution of LBW, FI n rinking wter ue to tretment with AlCl. This suggeste tht AA h Digestiility oeffiients n nutritive vlue: Tretment protetive effet ginst luminum toxiity. with AlCl use signifint (p<0.05) erese in Dry The erese in LBW, FI n DW of nimls Mtter (DM), Orgni Mtter (OM), Crue Protein (CP), trete with AlCl re in greement with the fining of Crue Fier (CF), Ether Extrt (EE), Nitrogen Free Extrt [27] [28] [29] Pettersen et l., Lloet et l. n Btineh et l.. (NFE), Totl Digestile Nutrients (TDN) n Digestile [0] Also, Alin et l. foun tht rits trete with Crue Protein (DCP) n use signifint (p<0.05) oses of luminium 25, 00 n 400 kgg showe erese inrese in nutritive rtio (NR) (Tle ). While, tretment in oy weight gin, espeilly with 400 mmol. Also, with AA lone use insignifint inrese in DM, OM, [] Gomez et l. foun tht tretment with luminium in CP, CF, EE, NFE, TDN n DCP n use signifint rinking wter t oses (50, 00 kgg yg ) for 6.5 month, (p<0.05) erese in NR. The presene of AA with AlCl use erese in oy weight of rts. Cherroret use insignifint inrese in DM, OM, CP, CF, EE, et [2] l. suggeste tht the reution in oy NFE, TDN n DCP n use insignifint (p<0.05) weight of trete young rts with luminium hlorie erese in NR. The men vlues of nitrogen lne (00 kgg yg ) for 5 to 4 ys oul e ttriute to the (NB), miroil el tivity (mmoni (NH-N) n erese in fee onsumption. Asori i use voltile ftty is (VFA) of mle rits (Tle 4). signifint inrese in oy weight, fee intke n Tretment with AlCl use signifint (p<0.05) erese in NB n use insignifint erese in NH-N n VFA. While, tretment with AA lone use insignifint Digestion oeffiients: DM 67.95± ± ± ± OM 68.85± ± ± ±.47 Items Control AA AlCl AA+AlCl CP 72.8± ± ± ±.48 Intke.46± ± ± ±0.0 EE 87.4± ± ± ±0.7 Fees 0.94± ± ± ±0.05 CF 7.00± ± ± ±.9 Urine 0.28± ± ± ±0.0 NFE 72.4± ± ± ±.5 Digeste 2.52± ±0.0.8± ±0.05 Nutritive vlue: TDN 67.7± ± ± ±.9 DCP.49± ± ±.5.66±0.0 NR.28±0.0.0± ± ±0.08 Mens within rows with ifferent supersript letters ifferent Signifintly (p<0.05) inrese in NB, NH-N n VFA. The presene of AA with AlCl use insignifint inrese in NB, NH -N n VFA. Anlysis of vrine showe tht nitrogen (intke, igeste n lne), NR n DCP were signifintly ffete y tretment (p<0.0). Also, EE ws signifintly ffete y tretment (p<0.05), while the other prmeters were not ffete. The present results gree well with Pettersen [27] [28] [29] et l., Lloet et l., Btineh et l. n Alin [0] et l. who reporte tht luminum use signifint Tle 4: Nitrogen lne n miroil el tivity of mle rits trete with sori i (AA), luminium hlorie (AlCl ) n their omintion (mens±se) Blne 2.24± ± ± ±0.06 Miroil tivity: NH-N (mg/00 ml) 5.2± ± ± ±0.5 VFA (meq/00 ml) 8.8± ± ± ±0.5 Mens within rows with ifferent supersript letters ifferent Signifintly (p<0.05) 2

4 Res. J. Agri. & Biol. Si., ():0-6, 2005 erese in oy weight, fee intke, weight gin n rinking wter. Bloo iohemil prmeters: Plsm trnsminses: Tretment with AlCl resulte in (Tle 5) signifint (p<0.05) inrese in the tivities of plsm sprtte minotrnsferse (AST) n lnine minotrnsferse (ALT). While, AA use signifint (p<0.05) erese in these enzymes s ompre with ontrol. The presene of AA with AlCl signifintly (p<0.05) erese the inution of AST n ALT tivities ue to tretment with AlCl n this mens tht AA llevite its toxiity. Trnsminses (AST n ALT) re importnt n ritil enzymes in the iologil [7] proesses. The inrese in plsm AST n ALT of nimls trete with AlCl (Tle 5) re greement with [8] the fining of Hssoun n Stohs, Chinoy n [9] [40] Memon n in El-Demersh. They foun tht exposure to AlCl use nerosis to the liver, AST is signifintly inrese in suh ses. It espes to the plsm from the injure hepti ells. Also, ALT level inites the existene of liver iseses, s this enzyme is present in lrge quntities in the liver. It inreses in serum when ellulr egenertion or estrution ours in this orgn. Therefore, the inrese of these enzymes in plsm re initive of liver mge n thus ltertions in liver funtion. The erese in plsm AST n ALT tivities of nimls trete with AA lone or in omintion with AlCl oul inite improvement of liver funtion n protetion from the toxiity of luminium. Plsm phosphtses: The overll mens of the tivities of plsm lkline phosphtse (ALP) n i phosphtse (ACP) were liste in (Tle 5). Dt inite tht tretment with AlCl use signifint (p<0.05) inrese in the tivities of plsm ALP n ACP. While, AA use signifint (p<0.05) erese in these enzymes ompre to ontrol nimls. The presene of AA with AlCl use signifint (p<0.05) erese in the inution of ALP n ACP tivities ue to tretment with AlCl n this mens tht AA h protetive effet ginst the toxiity of AlCl. Tle 5: Effet of sori i (AA), luminium hlorie (AlCl ) n their omintion on some plsm enzymes of mle rits (mens±se) Items Control AA AlCl AA+AlCl ALP (U LG ) 6.5 ± ± ± ± 0.60 GST (µmol hg ) 0.66 ± ± ± ± AST (U LG ) 5.8 ± ± ± ± 0.82 ALT (U LG ) 55.7 ± ± ± ± 0.94 ACP (U LG ) 0.9 ± ± ± ± 0.2 AChE (µmol ming ) 2.97 ± ± ± ± 0.05 TBARS (µmol ming ) 2.2 ± ± ± ± 0.0 Within row overll men with ifferent supersript letter iffer signifintly (p<0.05) Alkline phosphtse (ALP) enzyme is sensitive iomrker to metlli slts sine it is memrne oun [4] enzyme relte to the trnsport of vrious metolites. The tivity of ALP is onerne with energy metoli tivities n proesses in the oy n the erese in its tivity my inite impire energy proessing of [42] the ells. The inrese in the tivities of ALP n ACP of nimls trete with AlCl (Tle 5) re in orne [4] [44] with the finings of Szilgyi et l., El-See et l. n [6] [40] Ohmnski n Brsz. Also, El-Demersh foun tht the tivities of these enzymes were inrese in mie fe on whet ontining luminium resiue of 0.2 kgg [42] BW. Szilgyi et l. foun tht exposure to luminium use inrese the levels of ALP ue to inrese osteolsti tivity, provoke y the isturne of [6] one formtion. Ohmnski n Brsz reporte tht luminium my in to DNA, RNA n inhiit the tivities of i n lkline phosphtses. In ition, they reporte tht the inrese in the tivity of ALP or ACP in loo might e ue to the nerosis of liver, kiney n lung. The erese in plsm tivities of ALP n ACP ue to tretment with AA lone or in omintion with AlCl oul e inition to improve liver funtion n protetion from the toxiity of luminium. Plsm etylholinesterse: The effets of AA, AlCl n/or their omintion on plsm etylholinesterse (AChE) tivity uring the 6 weeks experimentl perio re shown in (Tle 5). Dt inite tht AlCl use signifint (p<0.05) erese in plsm n tivity ompre to ontrol nimls. While, AA lone i not hnge the tivity of AChE. The presene of AA with AlCl use signifint (p<0.05) inrese in the reution of plsm n rin AChE ue to tretment with AlCl n this mens tht AA llevite its toxi effet on the tivity of AChE. The inhiition of AChE tivity erese the ellulr metolism, inluing eformities of ell memrne n [45,46] isturne of metoli n nervous tivity. Also, the erese AChE tivity oul le to ioni [47] refluxes n ifferentil memrne permeility. [48] Suresh et l. reporte tht the erese in AChE tivity oul e ue to the erese of the enzyme synthesis y the inhiitory nture of toxint. Oviously, n impire neurohemil mehnism oul limit the musulr tivity of the niml, inluing suh vitl [45] funtions s respirtion, with lethl onsequenes. The erese (Tle 5) in AChE of nimls trete with [48] [49] AlCl is in greement with Srin et l., Dv et l. n [40] [50] El-Demersh. Moshtghie et l. reporte tht Al my interfere with either synthesis of etylholinesterse or inhiits holine uptke y synptosomes. They

5 Res. J. Agri. & Biol. Si., ():0-6, 2005 reporte tht the neurotoxiity of luminum my e s iomoleules, suh s lipis, DNA n proteins, hs [5] result of lipi peroxition. Nyk n Chtterjee een implite in mny hroni iseses, in prtiulr, reporte tht luminum inue umultion of [0] riovsulr isese, ner n trt. The glutmte or other ltertions in enzymes of the inrese in TBARS of nimls trete (Tle 6) with AlCl [56] [52] glutmte-gaba system my e one of the uses of is in greement with Frg et l., Ktyl et l. n luminum inue neurotoxiity. Moreover, Chinoy [57] Lll et l.. It ws foun tht lipi peroxition inue n [8] Memon referre the inhiition in y luminium t su-lethl levels, lter physiologil [58] etylholinesterse tivity to the effet of AlCl on n iohemil hrteristis of iologil systems. the synpti trnsmission. The erese in TBARS of nimls trete with AA is [59] in greement with Hung et l. who reporte tht Plsm glutthione S-trnsferse: The men vlues of sori i (500 mg sorte G ) supplementtion plsm glutthione S-trnsferse (GST) tivity s in nonsmokers reue lipi peroxition. Herzynskffete y tretment with AA, AlCl n/or their Cero et l. reporte tht supplementtion with [60] omintion throughout the 6 weeks experimentl perio sori i suppresses leukoyte oxygen free ril re presente in Tle 5. Tretment with AlCl resulte in proution in ptients with myoril infrtion. signifint (p<0.05) erese in the tivity of GST. Aitionlly, AA n protet iomemrnes ginst While, AA use signifint (p<0.05) inrese in peroxitive mge. plsm GST ompre to ontrol. The presene of AA with AlCl use signifint (p<0.05) inrese in the REFERENCES reution of GST tivity ue to tretment with AlCl n this mens tht AA llevite its toxi effet.. Sigel, H. n A.E. Sigel, 988. Metl ions in ilysis Glutthione S-trnsferses (GSTs) re fmily of emeni synrome n luminum intoxition. enzymes tht tlyze the ition of the tripeptie Nephron., : -0. glutthione to enogenous n xenoiotie sustrtes 2. Ageny for Toxi Sustnes n Disese Registry, whih hve eletrophili funtionl groups. They ply n 990. Toxiologil Profile for Aluminum. US importnt role in the etoxition n metolism of mny Deprtment of Helth n Humn Servis Puli xenoioti n enoioti ompouns. The erese in Helth Servies. GST (Tle 5) of nimls trete with AlCl is in greement. Jones, X.C. n B.G. Bennett, 986. Exposure of mn [52] [5] [40] with Ktyl et l., Du n Gill n El-Demersh. environmentl luminum-nexposure ommitment ssessment. Si. Totl Environ., 52: Thiorituri i-retive sustnes in plsm: The 4. Minmi, T., M. Ihii, Y. Tohno, S. Tohno et l., 996. overll mens of plsm thiorituri i-retive sustnes (TBARS) onentrtions (Tle 5). The t inite tht plsm TBARS signifintly (p<0.05) inrese y AlCl tretment. While, AA use signifint (p<0.05) erese in plsm TBARS s ompre with ontrol. The presene of AA with AlCl use signifint (p<0.05) erese the inution of TBARS in plsm ue to tretment with AlCl n this mens tht the presene of AA minimize the hzrous effet of AlCl. Free rils ttk ell strutures within the oy, using ell memrne, enzyme n DNA mge. As result, free rils hve een implite in numerous iseses, exmples of whih re: ner, onitions ssoite with premture irth, theroslerosis, motor neuron isese, rheumtoi rthritis, liver mge, ietes, respirtory isese, trts n entrl [54] nervous system isorers. Lipi peroxition is one of the min mnifesttions of oxitive mge n hs een foun to ply n importnt role in the toxiity of [55] mny xenoiotis. Also, oxitive mge to Age-epenent luminum umultion in the humn ort n ererl rtery. Biol. Tre. Elem. Res., 55: Bush, V.J., T.P. Moyer, K.P. Btts n J.E. Prisi, 995. Essentil n toxi element onentrtions in fresh n formul-fixe humn utppsy tissues. Clin. Chem., 4: Ohmnski, W. n W. Brsz, Aluminumourrene n toxiity for orgnisms. Przegl. Lek., 57: John, N.H., 982. Nutritionl Toxiology: Definition n Sope. In: (Nutritionl Toxiology, emi press, In.), : Yousef, M.I., F.M. EL-Demersh n E.I. EL-Agmy, 999. Effet of sori i on some iohemil prmeters of rits ffete y fltoxin B. Environ. Nut. Intl., : Slem, M.H., K.I. Kmel, M.I. Yousef, G.A. Hssn n F.D. EL-Nouty, 200. Protetive role of sori to enhne semen qulity of rits trete with sulethl oses of fltoxin B. Toxiology, 62:

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