The Reproductive Performance of Breeder Cocks Fed Cottonseed Cake-based Diets

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1 Interntionl Journl of Poultry Siene 6 (2): , 2007 ISSN Asin Network for Sientifi Informtion, 2007 The Reprodutive Performne of Breeder Coks Fed Cottonseed Cke-sed Diets G.O. Adeyemo, O.G. Longe nd D.O. Adejumo Deprtment of Animl Siene, University of Idn, Idn, Oyo Stte, Nigeri Astrt: Histology of the testes of reeder oks fed replement levels of 0, 25, 50, 75, or 100% of ottonseed mel (CSM) for soyen mel (SBM) in five different reeder diets for fifteen weeks were investigted in ompletely rndomized design. Chemil nlyses were rried out to determine the rude protein (CP) nd gossypol ontent of CSM. The 23 week-old reeder oks were fed the experimentl diets for 15 weeks, euthnized nd their testes removed for histologil studies. The determined CP nd gossypol ontents of CSM were 35.11% nd 570 g/ton respetively. The dily sperm prodution (DSP) y the reeder oks ws depressed when CSM dietry inlusion ws ove 50% with DSP vlues rnging from to /ml. The investigtion showed tht repling 75% SBM with CSM ws not injurious to the nonreprodutive helth of the reeder oks. However, CSM my not reple more thn 50% of SBM for reeder oks euse of the depressing effet of gossypol on sperm prodution when this level is exeeded. Key words: Breeder oks, ottonseed mel, dily sperm prodution, histology, reprodution Introdution determine the effet of CSC on reprodutive In the lte 1930 s nd erly 1940 s not single hildirth ws reported during period of ten yers in one re of Chin. It ws found tht for eonomi resons villgers performne of reeder oks nd the extent to whih it n e inorported into their diets without depressing their fertility. hd swithed from ooking with soyen oil to rude ottonseed oil (Liu, 1957). Women suffered from Mterils nd Methods menorrhe nd men were impotent. These Experimentl diets nd ird mngement: Five epidemiologil findings led to linil studies nd isonitrogenous nd isolori diets were formulted s further niml investigtions. Severl investigtors hve shown in Tle 1. The ontrol tretment (diet 1) reported the ntifertility effet of gossypol in mles nd ontined no CSM, while diets 2 to 5 were formulted y femles of non-ruminnt speies (Wu et l., 1981; sustituting CSM t 25, 50, 75, nd 100% for SBM Anderson, 1985; Bender et l., 1988). It ws olletively protein. indited tht gossypol eti id tretment of femle Thirty 23 week-old egg lying leghorn reeder oks rodents disrupts the norml pttern of estrous yles were rndomly llotted to five different dietry tretments through effet on pituitry nd ovrin hormone with three replites, eh hving 2 oks per replite. seretion. Post implnttion development disruption The irds were moved into the experimentl room t 21 ws further reported y the sme set of reserhers. weeks of ge nd llowed to limtize for two weeks It hs een reported tht t effetive doses, gossypol on the deep litter pens, fter whih they were weighed on uses mles to e infertile euse of sperm the dy the feeding tril strted. Susequent weighing immotility nd depressed sperm ounts (Rndels et l., ws reorded t weekly intervls. Wter nd feed were 1992). Speifi mitohondril dmge in the tils of provided d liitum nd from these reord of weekly spermtozo seems to render them immotile nd feed intke ws tken. From the reords of dily feed extensive dmge to germinl epithelium my e intke nd weight gin the feed onversion rtio ws responsile for depressions in spermtogenesis. lulted. The feeding tril ws rried out for twelve Findings hve lso shown tht there re mrked weeks. All niml re proedures were s pproved y differenes in speies suseptiility to gossypol toxiity the Animl Cre nd Use Unit of the University of Idn. (Kll et l., 1982; Weinuer et l., 1982; Wong et l., 1984). It hs een reported tht nti-fertility responses Testiulr histology: Testiulr density ws lulted re oth dose nd time dependent (Sksen nd Slmonsen, 1982). Gossypol tretment t puertl time upwrds mrkedly ffeted the reprodutive performnes of test nimls depending on the doses pplied. The ojetive of this reserh therefore ws to from the weight nd volume of the testes. The density 3 ws expressed in g/m entimeter. Smples from the left nd right testis of oks from eh tretment were fixed in out 10 times their volume in queous Bouin s solution for 6 h. Histologil setions of 7 mirons 140

2 Adeyemo et l.: The Reprodutive Performne of Breeder Coks Tle 1: Composition of experimentl otton seed ke sed diets fed to reeder Coks Tretments Ingredients (Kg) Mize Soyen mel Cottonseed ke Whet offl Fish mel Blood mel Lysine Methionine Oyster shell Bone mel Slt *Breeder premix Totl Clulted CP (%) ME (Kl/Kg) 2,806 2,809 2,801 2,800 2,804 *Premix supplied per kg of diet: Vit A, 10,000 IU; Vit D3, 2,800 IU; Vit E, 35,000 IU; Vit K, 1,900mg; Vit B12 19mg; Rioflvin, 7,000mg; Pyridoxine, 3,800mg; Thimine, 2,200mg; D- Pntotheni id, 11,000mg; Niotini id, 45,000 mg; Foli id, 1,400mg; Biotin, 113mg; Cu, 8,000mg; Mn, 64,000 mg; Zn, 40,000mg Fe, 32,000mg; Se, 160mg; Iodine, 800mg; Colt, 400mg; Choline, 475,000mg; Methionine, 50,000mg; BHT, 5,000mg; Spirmyin, 5,000mg Tle 2: Chemil omposition of experimentl reeder diets Prmeters (%) Diet 1 Diet 2 Diet 3 Diet 4 Diet 5 Dry mtter Crude protein Crude fire Ether extrt Ash Nitrogen free extrt Moisture Totl o thik were floted nd flttened out on 40 C wter nd then piked up with len slides smered with Myer s egg lumin. The slides mde were inuted for 30 min efore stining. Two sets of slides were prepred for eh ok. One set ws stined ording to the Period-Aid-Shiff (PAS) tehnique nd ounter stined with hemtoxylin. The seond set ws stined with hemtoxylin-eosin (H nd E). The stges of development of round spermtids to spermtozo were identified y omined method of rosome development ssessment s stted y PAS tehnique (Lelond nd Clermont, 1952) nd y nuler morphology shown y hemtoxylin-eosin stined prffin emedded tissue (DeReviers, 1971). The volumetri proportions of spermtogeni elements in the seminiferous epithelium were determined using the methods of (Chlkley, 1943). A twenty-five point oulr grtiule ws used in the estimtion. The dily sperm prodution (DSP) ws determined y quntittive testiulr histology. Determintion of dily sperm prodution (DSP) y quntittive testiulr histology: The DSP n e lulted from informtion sed on the morphometri nlysis of histologil setions. A method ws developed in 1970 whih utilized the volumetri proportions nd life spn of round spermtids in the tissue ross setion (Amnn, 1970). The ury of this method ws ensured y orreting for the shrinkge of the testis volume during histologil proesses. The time divisor lulted from the volume perent of the round spermtid is reported to e more urte thn tht for mture spermtids (Amnn, 1981). The DSP ws determined from the histologil nlysis using the eqution elow: DSP = (CTV) (vol % of round spermtid nulei in the testis%) / (Ave. vol per round spermtid nuleus in testis) (Life spn of round spermtids in dys). The orreted testiulr volume (CTV) ws otined y the formul CTV = (Gross testis weight-tuni lugine wtvolume, of meditinum%) (Shrinkge orretion) Shrinkge = Initil volume-finl volume / Initil orretion ftor volume This formul hs the dvntge of eing dptle for DSP lultions in vrious speies, if the life spn of round spermtids, verge volume of round nuleus, shrinkge vlue nd volumetri proportion of the speies re known. Sttistil nlysis: Dt otined from the prmeters onsidered were sujeted to desriptive sttistis nd nlysis of vrine using (SAS, 1999). Differenes etween mens were deteted using Dunn s multiple rnge tests. Results Identifition of spermtogeni elements nd ellulr ssoitions: Sperm ells were identified t their different stges of development following their retions to the stining tehniques used. Spermtogoni A line the lmin of the seminiferous tuules nd were hrterized y lrge ple ovoid nuleus rnging from 2.82 to 2.99 mirons in dimeter. Spermtogoni B were hrterized y smller nd drk nuleus hving dimeter rnge of 1.34 to 1.39 mirons, nd they were more nerly spheril in shpe. The sertoli ells were esily identified y their hrteristi irregulr shpes with deep invgintions of nuler memrne. They were seen to support the spermtogeni epithelium. The size of the sertoli ells rnged from 0.71 to 0.95 mirons. 141

3 Adeyemo et l.: The Reprodutive Performne of Breeder Coks Tle 3: Averge seminiferous tuulr dimeter nd volumetri proportion of testiulr elements of oks fed ottonseed ke seddiets Cell type (%) Diet 1 Diet 2 Diet 3 Diet 4 Diet 5 SEM Spermtogoni A Spermtogoni B Spermtoyte Spermtoyte Round Spermtid Elongted Spermtid Spermtozo Sertoli ells Interstitil ells Cytoplsm Tuulr Dimeter (µ) Mens within the sme row with different supersripts re signifintly different (p<0.05) signifint differenes reorded existed etween only tretment 5 nd the other tretments. Dily sperm prodution (DSP) in reeder oks fed ottonseed ke-sed diets: The results of DSP were sed on histometri nlysis nd presented in Fig. 1 elow. The highest DSP vlue ws reorded for tretment 1 (2.72) while the lowest vlue oserved ws from tretment 5 (1.48). There were differenes (p<0.05) etween the dietry tretments. Tretments 4 nd 5 were lower (p<0.05) thn the other tretments. Dily Sperm Prodution vlues fell shrply fter tretment 3 with the inlusion level of CSC t or greter thn 13%. Fig 1: 3 Dily sperm prodution (x10 ) vlues of reeder oks fed CSC-sed diets Primry spermtoytes looked like spermtogoni B ut with more visile nuleoli nd hromosome. The o primry (1 ) spermtoytes rnge from to mirons in dimeter. Seondry spermtoytes hve spheril nulei with network of hromtin grnules like the type oserved in the first stge of spermtid development, ut lrger thn the round spermtid. Their dimeter rnge in mirons vries from 0.22 to Volumetri proportions of testiulr elements: The results of the volumetri proportions of testiulr elements in reeder oks fed ottonseed ke sed diet re presented in Tle 3. Differenes (p<0.05) were oserved in most of the testiulr elements mesured. Spermtogoni A nd B did not show signifint differene. Interstitil ell ws highest in the ontrol (0.79%) nd lowest in tretment 4 (0.65%).The defining ell of testiulr tivities, the sertoli ells ws highest in the ontrol diet (0.95%) nd lowest in tretment 5 (0.71%). Tuulr dimeter of seminiferous tuules: The men tuulr dimeter is lso shown in Tle 3. The Disussion The results otined in this study re similr to the work of (Nkng, 1989) nd (Nkng nd Egunike, 1990) who identified two tegories of spermtogoni sed on their nuler size, hromtin onfigurtion nd the reltive position of these ells in the seminiferous tuules. This identifition nd similrity of the rrngement of ells is lso orroorted y erlier uthors, (Lelond nd Clermont, 1952; Clermont, 1960; Mldjin et l., 2002) who hve morphologilly differentited etween spermtogoni A nd B. The five tegories of primry spermtoytes reported y (DeReviers, 1971; Nkng nd Egunike, 1990) in oks were oserved in the testiulr setions mde from testis of the reeder oks used for this experiment. Though similrities were oserved in the five tegories of ells in ll the dietry tretments, signifint differenes were reorded s dietry levels of ottonseed ke protein replement for soyen exeeded 50%. Every speies hs its pity for sperm prodution whih is determined genetilly ut it hs een lerly oserved tht other ftors like nutrition, disese nd stress influene whih portion of the germinl epithelium enters into spermtogenesis (Grner nd Hfez, 1993; Ethu nd Egunike, 2002) reported tht poultry exudtes re very sensitive to dietry hnges 142

4 Adeyemo et l.: The Reprodutive Performne of Breeder Coks nd tht the slightest imlne in feed omposition testiulr weights were oserved in irds on diets 1 to 3 goes long wy in influening its physiology nd thn diets 4 nd 5. Testiulr weights hve een susequently its performne. Signifint differenes reported to hve high orreltion with sperm reserves were noted in some of the elements onsidered like in the testis or epididymis nd therefore refletion of primry spermtoytes, round spermtids, elongted sperm prodution. Thus the higher the testiulr weight, spermtids, seminiferous tuules, sertoli ells nd the more the sperm prodution. Additionlly, the testes spermtozo. weights were higher in the ontrol diets nd diets 2 nd The vlues otined in this work gree with the report of 3 euse they hd higher proportion of their testiulr (DeReviers, 1971; Nkng nd Egunike, 1990) nd re mss oupied y the seminiferous epithelium while the lose enough for omprison. From the results of this tuni lugine formed higher proportion of the work it ws glring tht ny diet tht inreses the level testiulr mss in diets 4 nd 5. This influened the of nti-nutritive ftors prtiulrly free gossypol eyond overll weight of the testis whih ws refleted in the the mount in diet 3 (0.01 g/ton) would disourge the mount of sperm produed dily y the reeders. prodution of germ ells nd susequently From the results oserved, CSC my not reple more spermtogoni. Dietry tretments tht hd higher proportions of spermtozo were lso oserved to hve high orresponding proportion of sertoli ells. The implition of this is tht ny sustne tht limits the tivity of sertoli ells will dversely ffet sperm thn 50% of SBC for reeder oks euse of its depressing effet on ok fertility. Thus CSC n effetively sustitute SBC up to 50% y providing the needed protein nd qulity feed essentil in the prodution performne of reeder oks. prodution. Dietry tretments tht hd higher thn 50% ottonseed Referenes ke (represented y ny free dietry gossypol greter Amnn, R.P., Sperm prodution rtes. In: the testis, eds. A.D. Johnson, W.R. Gomes nd N.L.I. thn 0.001%) replement for soyen ke hd Vndemrk, NY. Ademi Press, pp: signifintly lower sertoli ells vlues thn those with Amnn, R.P., A ritil review of methods for lower levels of ottonseed ke. The reltionship evlution of spermtogenesis from Seminl oserved etween sertoli ells nd spermtozo n e hrteristis. J. Andrology, 2: explined in reltion to the funtion of the sertoli ells Anderson, P.A., Intertions etween proteins nd whose ronym is nurse ell inpittion of the onstituents tht ffet protein qulity. In: nurse ell y nti-nutrient tivities mens the Digestiility nd mino id vilility in erels nourishment provided during the morphologil nd oil seeds (J.W. Finley nd D.T. Hopkins, eds.) trnsformtion of spermtids during spermiogenesis is Am. Asso. of Cerel Chemist. St. Pul, Minnesot, impeded whih susequently lowers sperm prodution The volume in (%) oupied y the seminiferous tuules Bender, H.S., G.K. Sunders nd H.P. Misr, A showed signifint tretment effet, nd the figures histopthologi study of the effets of gossypol on otined were lower thn those oserved y (DeReviers, the femle rt. Contreption, 38: ) for dult hikens. The volume (%) of Chlkley, H.W., Methods for quntittive seminiferous tuules in the testis hs een reported to morphologi nlysis of tissues. J. Ntl. Cner e speies speifi (Nkng, 1989). The seminiferous Inst., 4: tuule dimeters for the different diets follow the Clermont, Y., Cyle of the seminiferous epithelium proportion of sertoli ells in their tuules. It n e of the guine pig. A method for identifition of the inferred from the oserved vlues tht ny diet tht influenes the numer of sertoli ells will influene the size of the seminiferous tuules. Signifint differenes were oserved in DSP y the histometri method employed for this estimtion. These results, though slightly different, gree with the work of (Mldjin et l., 2002; Grner nd Hfez, 1993; Egunike nd Oluyemi, 1979). The slight vritions in figures re proly due to differenes in speies whih grees with the report of (Grner nd Hfez, 1993), tht every speies hs it s pity for sperm prodution whih is genetilly determined, though environmentl influenes nnot e ruled out, whih determines wht proportion of the germinl epithelium enters into spermtogenesis. The ontrol diet (diet 1) nd diets 2 nd 3 hd higher DSP vlues thn diets 4 nd 5 nd susequently, higher stges of growth. Fertil Steril, 11: DeReviers, M., Less development testiulr hez le oq. III. Influene de l duree Quotidiene delirement sous photoperiods onstnts. Am. Biol. Anim. Biohem. Biophys., 14: Egunike, G.N. nd J.A. Oluyemi, Comprtive studies of the reprodutive pity of Nigeri nd Exoti Poultry reeds. 1 Testis nd semen hrteristis Nig. J. Anim. Prod., 6. Ethu, K.A. nd G.N. Egunike, Effet of proessed sweet potto on the performne of roiler irds in the humid tropis during the riny seson. Trop. Anim. Prod. Invest., 5: Grner, D.L. nd E.S.E. Hfez, Spermtozo nd seminl plsm. In: Reprodution in Frm Animls Ed. E.S.E. Hfez 6th Le nd Feiger, Phildelph. 143

5 Adeyemo et l.: The Reprodutive Performne of Breeder Coks Kll, N.R., T.W. Foo nd A.R. Sheth, Studies on Rndels, R.D., C.C. Chse, Jr. nd S.Y. Wys, the mle ntifertility gent-gossypol eti id on Effets of gossypol nd ottonseed produts on the fertility of mle rts. Andrologi, 14: 492. reprodution of mmmls. J. Anim. Si., 70: Lelond, C.P. nd Y. Clermont, Spermtogenesis of rt, mouse, hmster, nd Guine pig s reveled Sksen, S.K. nd R.A. Slmonsen, Anti-fertility of y the periodi id-funtion sulfurous id gossypol in mle hmsters. Fertil. Steril., 37: 686. tehnique Am. J. nt., 90: SAS Institute, SAS User Guide: Sttistis. Liu, B.S., A tenttive ide of the use of ooking Version 9 Edition. SAS Institute, In.,Cry, NC. U.S.A. ottonseed oil for fertility ontrol. Shnghi J. Chin. Weinuer, G.F., E. Rovn nd J. Frik, Antifertility Med., 6: 43. effiy of gossypol eti id in mle rts. Mldjin, A., P. Penny nd R. Nole, DHA-rih Andrologi, 14: 270. mrine oils nd improved reprodutive effiieny in Wong, P.Y.D., J.P. Qiu nd W.O. Fu, Gossypol pigs. Annul Meeting of the Amerin Oil Soiety does not ffet testiulr fluid seretion in rts. (AOCS), Montrel, Cnd, ook of strts. Contreption, 29: 471. Nkng, E.E. nd G.N. Egunike, Cytology nd the yle of the seminiferous Epithelium in the ok. Wu, Y.M., S.C. Chppel nd G.L. Flikinger, Effets Trop. Anim. Prod. Invest., 1: of gossypol on pituitry-ovrin endorine funtion, Nkng, E.E., Breed nd Sesonl Influenes on ovultion nd fertility in femle hmsters. the reprodutive potentil of the Cok in humid Contreption, 24: 259. Tropil environment. Ph.D thesis. Animl Siene Deprtment, University of Idn, Idn, Nigeri. 144