Olive pulp and exogenous enzymes feed supplementation effect on the carcass and offal in broilers: a preliminary study

Ag icul u e 2020, 10, 359; doi:10.3390/ag icul u e10080359 www.mdpi.com/jou nal/ag icul u e A icle Oli e Pulp and Exogenous Enzymes Feed Supplemen a ion E ec on he Ca cass and O al in B oile s: A P elimina y S udy Peyman Sayehban 1, Ali eza Seida i 1,*, Mohammad Dadashbeiki 2, Ahmad Gho bani 3, Wagne Azis Ga cia de A aújo 4, Alessand a Du azzo 5, Massimo Luca ini 5, Paolo Gab ielli5, Besma Om i 6,7, Luiz Fe nando Teixei a Albino 8, Eliana B. Sou o 9,10 and An onello San ini 11,* 1 Depa men o Animal Science, Rash B anch, Islamic Azad Uni e si y, Rash 41335-3516, I an; [email p o ec ed] 2 Depa men o Ve e ina y Science, Rash B anch, Islamic Azad Uni e si y, Rash 41335-3516, I an; dadashb[email p o ec ed] 3 Guilan Ag icul u al and Na u al Resou ces Educa ion and Resea ch Cen e , Rash , 14445-41996, I an; [email p o ec ed] 4 Ins i u o de Educação, Ciência e Tecnologia No e de Minas Ge ais; 39480-000 Januá ia, MG, B azil; [email p o ec ed] 5 CREA-Resea ch Cen e o Food and Nu i ion, Via A dea ina 546, 00178 Rome, I aly; [email p o ec ed] (A.D.); massimo.luca ini@c ea.go .i (M.L.); [email p o ec ed]o .i (P.G.) 6 Labo a o y o Imp o emen and In eg a ed De elopmen o Animal P oduc i i y and Food Resou ces, Depa men o Animal P oduc ion, Highe School o Ag icul u e o Ma eu , Uni e si y o Ca hage, 1054 Ca hage, Tunisia; [email p o ec ed] 7 Na ional Ag onomy Ins i u e-Tunis, Uni e si y o Ca hage, 1054 Ca hage, Tunisia; 8 Depa men o Animal Science, Uni e sidade Fede al de Viçosa; 36570-000 Viçosa, MG, B azil; [email p o ec ed] 9 Depa men o Pha maceu ical Technology, Facul y o Pha macy, Uni e si y o Coimb a (FFUC), Pólo das Ciências da Saúde, 3000-548 Coimb a, Po ugal; [email p o ec ed] 10 CEB - Cen e o Biological Enginee ing, Uni e si y o Minho, Campus de Gual a 4710-057 B aga, Po ugal 11 Depa men o Pha macy, Uni e si y o Napoli Fede ico II, Via D. Mon esano 49, 80131 Napoli, I aly * Co espondence: ali ezaseida i@iau ash .ac.i (A.S.); Tel.: +98-911-331-3073; [email p o ec ed] (A.S.); +39-81-253-9317 Recei ed: 16 July 2020; Accep ed: 12 Augus 2020; Published: 14 Augus 2020 Abs ac : Nowadays, he e is an inc easing in e es in he exploi a ion and alo iza ion o ag icul u al ood was e and by-p oduc s. A he same ime, he g owing demand by ma ke s wo ldwide, especially in A ica and Sou heas Asia, can jus i y he g owing in e es in he use o by- p oduc s o he poul y indus y. Oli e pulp is one o he mos in e es ing by-p oduc s o oli e ee a ming ( ypical o he Medi e anean a ea), being a good sou ce o many biologically ac i e compounds wi h an ioxidan , an i ungal, and an ibac e ial p ope ies. The p esence o p ocessed oli e pulp in he die showed o be e ec i e in inc easing he weigh o speci ic ca cass and o al ai s. This wo k aims a s udying oli e pulp as a eed supplemen in poul y nu i ion, by ocusing on he e ec s on b oile ca cass and o al. Oli e pulp (OP) is one o he by-p oduc s o oli e ee a ming, being he esidue o oli e cake a e i is d ied. To e alua e he e ec s o OP in a die supplemen ed wi h di e en le els o a comme cial enzyme (ENZ) blend on b oile ca cass and o al ai s, h ee hund ed male b oile chicks (Ross 308 lineage; one-day-old) we e di ided in o en ea men g oups acco ding o a comple ely andomized design. The ea men s die s con ained: unp ocessed OP (50 g/kg, 100 g/kg, 50 g/kg wi h ENZ, 100 g/kg wi h ENZ), p ocessed OP (50 g/kg, 100 g/kg, 50 g/kg wi h ENZ, 100 g/kg wi h ENZ), and con ol g oups (wi hou OP, and wi hou OP Ag icul u e 2020, 10, 359 2 o 17 wi h ENZ). The OP p ocessing inc eased b eas pe cen ages in b oile s. Supplemen a ion wi h ENZ did no change any o he s udied ca cass o o al ai alues. The p esence o OP (50 g/kg) in b oile die s inc eased he e isce a ed ca cass, leg, and neck pe cen age alues. The p esence o p ocessed OP (50 g/kg) in he die showed o be e ec i e in inc easing he weigh o speci ic ca cass and o al ai s. Keywo ds: by-p oduc s; eed p ocessing; eeds u s; oli e pulp; poul y; supplemen s. 1. In oduc ion Oli e pulp (OP) is he emainde o oli e cake ( aw ma e ial esul ing om oli e oil ex ac ion) a e he cake is d ied. I is also one o he mos in e es ing by-p oduc s o oli e ee a ming, being a good sou ce o se e al biologically ac i e compounds wi h an ioxidan , an i ungal, and an ibac e ial p ope ies [1–5] wi h a g ea nu aceu ical po en ial [6–14]. The use o by-p oduc s om di e en ege al o igins o supplemen eed is, on he o he hand, a ac ing g owing in e es due o en i onmen ally iendly use, e icacy, and sus ainabili y, o he han a oiding he necessi y o dump po en ially use ul and aluable by-p oduc s o he ag o ood sys em [15,16]. None heless, a en ion should be gi en o po en ial seconda y me aboli es p esen in he oli e [17–20] adop ing he necessa y p ecau ion in he ield o a oid hei p esence [21]. The OP is conside ed a good sou ce o p o ein, a , calcium, coppe , and cobal , bu i is low in i s nu i i e alue (ene gy, diges ible p o eins, and mine als) and i has a high lignin con en . I is also poo in some me als, e.g., phospho us, magnesium, and sodium, bu i con ains easonable le els o manganese and zinc [22–24]. The ipening s age a ha es in e e es wi h pec ic polysaccha ides, which a e ound in he OP cell walls due o he p esence o calcium chela ing dime s, he eby changing he nu i ional alue o his by-p oduc [25].Whe eas in he pas , he use o c op esidues and by-p oduc s as al e na i es o ce eals–soybean meal-based a ions o b oile s die was no success ul, mainly due o he high ibe con en and poo diges ibili y, now i is inc easing in in e es [26–28]. Some exogenous enzymes may be added o b oile die s con aining hese by-p oduc s as an aid o ibe diges ion (ca bohyd ases) o phy ic phospho us solubiliza ion (phy ase), he eby educing hei nega i e e ec s on b oile p oduc ion [29]. La elli and Bondesan [30] obse ed an inc ease in he o al secoi idoid polyphenol (an ioxidan , an imic obial, and an i-in lamma o y compounds) con en and an ioxidan ac i i y in ex a i gin oli e oil when he ui s we e p e- des oned. Mos o hese compounds a e e ec i e an ioxidan s; acco ding o Kidd [31], an ioxidan subs ances can educe cellula ee adical damage and imp o e he b oile s’ immunology, pe o mance, and ca cass. A ecen s udy by Debbou-Iouknane e al. [32] in es iga ed he in i o an icoccidial e ec o oli e pulp (Olea eu opaea L a . Chemlal) ex ac on he des uc ion o Eime ia spp. Oocys s isola ed om in ec ed chickens. The indings o his s udy showed ha phenolic compounds o OP ex ac es ed sepa a ely possess an an i-Eime ia spp. e ec . The ecen s udy o Papadomichelakis e al. [33] on he e ec s o die a y d ied oli e pulp inclusion on g ow h pe o mance and mea quali y o b oile chickens, showed ha b oile chickens u ilize d ied oli e pulp (DOP) supplemen ed die s mo e e icien ly when die a y DOP inclusion is inc eased g adually wi h age, i.e. by using a combina ion o g owe and inishe die s wi h a maximum o 25 and 50 g DOP/kg, espec i ely. The die a y addi ion o oli e cake o b oile s up o he le el o 150g/kg did no a ec pe o mance pa ame e s [34]. Za ei e al. [35] also epo ed ha he inclusion o up o 86g/kg o oli e pulp in he die o laying hens had no nega i e e ec s on p oduc ion pa ame e s. O he esea che s ound posi i e e ec s o he nu i ional use o oli e pulp. Abo Oma [36] epo ed an inc ease in b oile eed in ake (and a dec ease in eed e iciency) wi h he inclusion o abou 60g o oli e pulp/kg die . This au ho ela ed his high eed in ake o he ibe con en o he oli e pulp and he consequen inc ease in passage a e in he gas oin es inal ac . Howe e , he die a y addi ion o 75g/kg o OP has a nega i e e ec on weigh gain, acco ding o Rabayaae al. [37]. On he o he hand, he easibili y o including oli e pulp up o he le el o 160g/kg in b oile die s was epo ed [38]. The Ag icul u e 2020, 10, 359 3 o 17 use o enzymes o imp o e he nu i ional alue o oli e pulp was s udied in laying hens, bu no posi i e e ec s we e obse ed on p oduc ion o egg quali y pa ame e s [39]. Conside ing his con ex , u he s udies e alua ing he ca cass and o al o comme cial b oile s ed die s wi h OP seems o be needed, and o his eason, he objec i e o his p elimina y s udy was o assess he e ec o di e en die a y le els o p ocessed and unp ocessed OP wi h enzyme supplemen a ion on ca cass and o al o b oile chickens du ing a 6-week ial. 2. Ma e ial and Me hods Expe imen al p o ocols we e app o ed by he Animal Ca e Commi ee o he Islamic Azad Uni e si y, I an (p ocess #17–33–5–9013; 93–12–7). Th ee hund ed day-old Ross 308 male b oile s (A iagen; New B idge, Sco land, UK; 35805) we e di ided in o 30 g oups o 10 bi ds. Each cage (10 chickens) was assigned o a speci ic die a y ea men g oup. The expe imen al design had a o al o 10 ea men s (10 bi ds/ ea men ), and h ee eplica es pe ea men , as ollows:  50 p: p ocessed OP (50 g/kg) wi hou enzyme blend;  50 p + ENZ: p ocessed OP (50 g/kg) wi h enzyme blend (50 mg/kg);  100 p: p ocessed OP (100 g/kg) wi hou enzyme blend;  100 p + ENZ: p ocessed OP (100 g/kg) wi h enzyme blend (50 mg/kg);  50 u: unp ocessed OP (50 g/kg) wi hou enzyme blend;  50 u + ENZ: unp ocessed OP (50 g/kg) wi h enzyme blend (50 mg/kg);  100 u: unp ocessed OP (100 g/kg) wi hou enzyme blend;  100 u + ENZ: unp ocessed OP (100 g/kg) wi h enzyme blend (50 mg/kg);  C l: con ol die wi hou OP and wi hou enzyme blend;  C l + ENZ: con ol die wi hou OP wi h enzyme (50 mg/kg) blend. The bi ds we e housed in cages (dimensions: 1.25 × 1.25 m; loo a ea: 0.15 m2 pe bi d), which we e loca ed in a he mos a ically-con olled cu ain side-wall poul y ba n. The cage loo s we e co e ed wi h pape oll li e , and he bi ds emained in he cages o he du a ion o he expe imen , which ended a hei 42nd day o age. A wo-phase eeding schedule, which consis ed o s a e (1– 21 days) and g owe (22–42 days) eed pe iods, was adop ed in his expe imen al s udy. Table 1 and Table 2 epo he ing edien s and chemical composi ion o die s used du ing he s a e pe iod and he inishing pe iod, espec i ely. The die s me o exceeded he Ross 308 ca alog ecommenda ions o he s a e and g owe phases [40]. Ag icul u e 2020, 10, 359 4 o 17 Table 1. Feed ing edien s and chemical composi ion o die s used du ing he s a e pe iod (1s –21s days o age, as- ed basis). P ocessed Oli e Pulp (OPp) Unp ocessed Oli e Pulp (OPu) 503 p 4 50 p + ENZ 5 100 p 100 p + ENZ 50 u 6 50 u + ENZ 100 u 100 u + ENZ C l 7 C l +ENZ Ing edien s (g/kg) P ocessed OP 50.00 50.00 100.00 100.00 0.00 0.00 0.00 0.00 0.00 0.00 Unp ocessed OP 0.00 0.00 0.00 0.00 50.00 50.00 100.00 100.00 0.00 0.00 Enzyme 0.00 0.05 0.00 0.05 0.00 0.05 0.00 0.05 0.00 0.05 Co n 507.30 507.30 456.60 456.60 482.50 482.50 407.00 407.00 558.00 558.00 Soybean meal 370.60 370.60 370.60 370.60 377.20 377.20 383.70 383.70 370.70 370.70 Soybean oil 30.00 30.00 32.10 32.10 47.60 47.60 67.40 67.40 27.80 27.80 Whea b an 0.10 0.05 0.10 0.05 0.10 0.05 0.10 0.05 0.10 0.05 Dicalcium phospha e 19.30 19.30 19.60 19.60 19.40 19.40 19.70 19.70 19.00 19.00 Limes one 10.90 10.90 9.10 9.10 11.50 11.50 10.30 10.30 12.70 12.70 Vi amin mix u e 1 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 Mine al mix u e 2 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 Sal 2.30 2.30 2.10 2.10 2.40 2.40 2.40 2.40 2.50 2.50 Sodium bica bona e 1.50 1.50 1.50 1.50 1.50 1.50 1.50 1.50 1.50 1.50 DL-Me hionine 1.40 1.40 1.50 1.50 1.40 1.40 1.50 1.50 1.30 1.30 Lysine hyd ochlo ide 0.60 0.60 0.80 0.80 0.40 0.40 0.40 0.40 0.40 0.40 To al 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 Nu ien analysis (g/kg) D y ma e 903.20 903.20 904.90 904.90 904.80 904.80 908.20 908.20 901.50 901.50 Me abolizable ene gy (kcal/kg) 3.025 3.025 3.025 3.025 3.025 3.025 3.025 3.025 3.025 3.025 C ude p o ein 230.00 230.00 230.00 230.00 230.00 230.00 230.00 230.00 230.00 230.00 E he ex ac 59.00 59.00 65.20 65.20 74.70 74.70 96.80 96.80 52.70 52.70 Linoleic acid 27.90 27.90 27.90 27.90 36.40 36.40 44.80 44.80 27.90 27.90 C ude ibe 45.80 45.80 64.90 64.90 50.70 50.70 74.70 74.70 26.70 26.70 Calcium 10.50 10.50 10.50 10.50 10.50 10.50 10.50 10.50 10.50 10.50 Phospho us 7.40 7.40 7.30 7.30 7.30 7.30 7.30 7.30 7.40 7.40 A ailable phospho us 5.00 5.00 5.00 5.00 5.00 5.00 5.00 5.00 5.00 5.00 Po assium 9.50 9.50 9.90 9.90 9.40 9.40 9.50 9.50 9.20 9.20 Chlo ine 1.80 1.80 1.80 1.80 1.90 1.90 1.90 1.90 1.90 1.90 Manganese (mg/kg) 474.27 474.27 474.11 474.11 475.36 475.36 476.25 476.25 474.44 474.44 Sodium 0.16 0.16 0.16 0.16 0.16 0.16 0.16 0.16 0.16 0.16 Zinc (mg/kg) 383.69 383.69 383.21 383.21 385.60 385.60 387.01 387.01 384.17 384.17 Ag icul u e 2020, 10, 359 5 o 17 Choline (mg/g) 1.59 1.59 1.56 1.56 1.59 1.59 1.56 1.56 1.62 1.62 Folic acid (mg/kg) 2.19 2.19 2.18 2.18 2.21 2.21 2.20 2.20 2.21 2.21 A ginine 14.80 14.80 14.60 14.60 14.90 14.90 14.90 14.90 15.00 15.00 Glycine 9.20 9.20 9.10 9.10 9.30 9.30 9.20 9.20 9.40 9.40 Se ine 11.00 11.00 10.90 10.90 11.10 11.10 11.00 11.00 11.20 11.20 Gly + Se 20.20 20.20 20.00 20.00 20.40 20.40 20.20 20.20 20.60 20.60 His idine 5.90 5.90 5.80 5.80 5.90 5.90 5.80 5.80 6.00 6.00 Isoleucine 9.30 9.30 9.20 9.20 9.40 9.40 9.30 9.30 9.40 9.50 Leucine 18.90 18.90 18.40 18.40 18.90 18.90 18.40 18.40 19.40 19.40 Lysine 12.70 12.70 12.70 12.70 12.70 12.70 12.70 12.70 12.70 12.70 Me hionine 4.70 4.70 4.70 4.70 4.70 4.70 4.70 4.70 4.70 4.70 Cys eine 3.60 3.60 3.60 3.60 3.60 3.60 3.60 3.60 3.70 3.70 Me + Cys 8.30 8.30 8.30 8.30 8.30 8.30 8.30 8.30 8.40 8.40 Phenylalanine 10.60 10.60 10.40 10.40 10.60 10.60 10.50 10.50 10.80 10.80 Ty osine 8.70 8.70 8.60 8.60 8.80 8.80 8.70 8.70 8.90 8.90 Phe + Ty 19.30 19.30 19.00 19.00 19.40 19.40 19.20 19.20 19.70 19.70 Th eonine 8.40 8.40 8.30 8.30 8.40 8.40 8.30 8.30 8.50 8.50 T yp ophan 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 Valine 10.20 10.20 10.10 10.10 10.30 10.30 10.10 10.10 10.40 10.40 1Vi amin A: 3,600,000 IU/kg; Vi amin D3: 800,000 IU/kg; Vi amin E: 7200 IU/kg; Vi amin K3: 800 mg/kg; Vi amin B1: 720 mg/kg; Vi amin B2: 2640 mg/kg; Vi amin B3 (Calcium pan o hena e): 4000 mg/kg; Vi amin B5 (Niacin): 12,000 mg/kg; Vi amin B6: 1200 mg/kg; Vi amin B9 (Folic acid): 400 mg/kg; Vi amin B12: 6 mg/kg; Vi amin H2 (Bio in): 40 mg/kg; Choline: 100,000 mg/kg; An ioxidan : 40,000 mg/kg; Excipien : 1 mg/kg. 2 Mn: 39,680 mg/kg; Fe: 20,000 mg/kg; Zn: 33,880 mg/kg; Cu: 4000 mg/kg; I: 400 mg/kg; Se: 80 mg/kg; Choline: 100,000 mg/kg; Excipien : 1 mg/kg.; 3 OP=50 g/kg.; 4 Die wi h OPp; 5 Die wi h ENZ; 6 Die wi h OPu; 7 Con ol die wi hou OP. Table 2. Feed ing edien s and chemical composi ion o die s used du ing he inishing pe iod (22nd–42nd days o age, as- ed basis). P ocessed Oli e Pulp (OPp) Unp ocessed Oli e Pulp (OPu) 503 p 4 50 p + ENZ 5 100 p 100 p + ENZ 50 u 6 50 u + ENZ 100 u 100 u + ENZ C l 7 C l +ENZ Ing edien s (g/kg) P ocessed OP 50.00 50.00 100.00 100.00 0.00 0.00 0.00 0.00 0.00 0.00 Unp ocessed OP 0.00 0.00 0.00 0.00 50.00 50.00 100.00 100.00 0.00 0.00 Enzyme 0.00 0.05 0.00 0.05 0.00 0.05 0.00 0.05 0.00 0.05 Co n 547.60 547.60 496.80 496.80 522.60 522.60 447.10 447.10 598.20 598.20 Soybean meal 323.20 323.20 323.20 323.20 329.80 329.80 336.30 336.30 323.30 323.30 Soybean oil 42.30 42.30 44.50 44.50 60.00 60.00 79.80 79.80 40.20 40.20 Whea b an 0.10 0.05 0.10 0.05 0.10 0.05 0.10 0.05 0.10 0.05 Dicalcium phospha e 17.00 17.00 17.30 17.30 17.10 17.10 17.40 17.40 16.70 16.70 Limes one 8.70 8.70 6.90 6.90 9.30 9.30 8.20 8.20 10.50 10.50 Vi amin mix u e1 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 Mine al mix u e2 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 3.00 Sal 2.30 2.30 2.10 2.10 2.50 2.50 2.40 2.40 2.50 2.50 Ag icul u e 2020, 10, 359 6 o 17 Sodium bica bona e 1.50 1.50 1.50 1.50 1.50 1.50 1.50 1.50 1.50 1.50 DL-Me hionine 1.10 1.10 1.20 1.20 1.10 1.10 1.20 1.20 1.00 1.00 Lysine hyd ochlo ide 0.20 0.20 0.40 0.40 0.00 0.00 0.00 0.00 0.00 0.00 To al 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 1000.0 Nu ien analysis (g/kg) D y ma e 90.36 90.36 90.53 90.53 90.52 90.52 90.85 90.85 90.19 90.19 Me abolizable ene gy (kcal/kg) 3150.0 3150.0 3150.0 3150.0 3150.0 3150.0 3150.0 3150.0 3150.0 3150.0 C ude p o ein 21.00 21.00 21.00 21.00 21.00 21.00 21.00 21.00 21.00 21.00 E he ex ac 7.24 7.24 7.87 7.87 8.82 8.82 11.02 11.02 6.62 6.62 Linoleic acid 3.49 3.49 3.49 3.49 4.34 4.34 5.18 5.18 3.49 3.49 C ude ibe 4.49 4.49 6.39 6.39 4.97 4.97 7.38 7.38 2.58 2.58 Calcium 0.90 0.90 0.90 0.90 0.90 0.90 0.90 0.90 0.90 0.90 Phospho us 0.68 0.68 0.68 0.68 0.68 0.68 0.67 0.67 0.68 0.68 A ailable phospho us 0.45 0.45 0.45 0.45 0.45 0.45 0.45 0.45 0.45 0.45 Po assium 0.87 0.87 0.90 0.90 0.85 0.85 0.87 0.87 0.84 0.84 Chlo ine 0.18 0.18 0.17 0.17 0.18 0.18 0.18 0.18 0.19 0.19 Manganese (mg/kg) 471.74 471.74 471.58 471.58 472.83 472.83 473.72 473.72 471.91 471.91 Sodium 0.16 0.16 0.16 0.16 0.16 0.16 0.16 0.16 0.16 0.16 Zinc (mg/kg) 381.73 381.73 381.25 381.25 383.63 383.63 385.05 385.05 382.21 382.21 Choline (mg/g) 1.48 1.48 1.45 1.45 1.48 1.48 1.46 1.46 1.51 1.51 Folic acid (mg/kg) 2.04 2.04 2.02 2.02 2.05 2.05 2.05 2.05 2.06 2.06 A ginine 1.33 1.33 1.31 1.31 1.34 1.34 1.34 1.34 1.35 1.35 Glycine 0.84 0.84 0.83 0.83 0.84 0.84 0.84 0.84 0.86 0.86 Se ine 1.00 1.00 0.99 0.99 1.01 1.01 1.00 1.00 1.02 1.02 Gly + Se 1.84 1.84 1.82 1.82 1.85 1.85 1.84 1.84 1.88 1.88 His idine 0.54 0.54 0.53 0.53 0.54 0.54 0.53 0.53 0.55 0.55 Isoleucine 0.84 0.84 0.83 0.83 0.85 0.85 0.84 0.84 0.85 0.85 Leucine 1.75 1.75 1.71 1.71 1.75 1.75 1.70 1.70 1.80 1.80 Lysine 1.11 1.11 1.11 1.11 1.11 1.11 1.11 1.11 1.11 1.11 Me hionine 0.42 0.42 0.42 0.42 0.42 0.42 0.42 0.42 0.42 0.42 Cys eine 0.33 0.33 0.32 0.32 0.33 0.33 0.32 0.32 0.34 0.34 Me + Cys 0.75 0.75 0.74 0.74 0.75 0.75 0.74 0.74 0.76 0.76 Phenylalanine 0.96 0.96 0.95 0.95 0.97 0.97 0.96 0.96 0.98 0.98 Ty osine 0.79 0.79 0.78 0.78 0.80 0.80 0.79 0.79 0.81 0.81 Phe + Ty 1.75 1.75 1.73 1.73 1.77 1.77 1.75 1.75 1.79 1.79 Th eonine 0.76 0.76 0.75 0.75 0.76 0.76 0.76 0.76 0.77 0.77 T yp ophan 0.27 0.27 0.27 0.27 0.27 0.27 0.27 0.27 0.27 0.27 Valine 0.94 0.94 0.92 0.92 0.94 0.94 0.93 0.93 0.95 0.95 1Vi amin A: 3,600,000 IU/kg; Vi amin D3: 800,000 IU/kg; Vi amin E: 7200 IU/kg; Vi amin K3: 800 mg/kg; Vi amin B1: 720 mg/kg; Vi amin B2: 2640 mg/kg; Vi amin B3 (Calcium pan o hena e): 4000 mg/kg; Vi amin B5 (Niacin):12,000 mg/kg; Vi amin B6: 1200 mg/kg; Vi amin B9 (Folic acid): 400 mg/kg; Vi amin B12: 6 mg/kg; Vi amin H2 (Bio in): 40 mg/kg; Choline: 100,000 mg/kg; An ioxidan : 40,000 mg/kg and 1 mg/kg Excipien . 2 Mn: 39,680 mg/kg; Fe: 20,000 mg/kg; Zn: 33,880 mg/kg; Cu: 4000 mg/kg; I: 400 mg/kg; Se: 80 mg/kg; Choline: 100,000 mg/kg; Excipien : 1 mg/kg.; 3 OP=50 g/kg;.4Die wi h OPp; 5Die wi h ENZ; 6Die wi h OPu; 7 Con ol die wi hou OP. Ag icul u e 2020, 10, 359 7 o 17 The OP was ob ained by washing esh oli e ui wi h wa e and milling hem. Table 3 epo s he chemical composi ion o wo ypes o oli e meal used in he expe imen al p ocedu es. A e hese s eps, he oli es we e added o ho wa e (80 °C) and cen i uged. A his s age, he wa e –oil emulsion was emo ed om he milled oli e, which we named “oli e cake” (OC). In he nex s ep, - ocophe ol (an ioxidan ) and an an i ungal oxin-binde (zeoli e adso ben ) we e added o he OC, which was hen d ied wi h ho ai (70 °C), he eby esul ing in he OP. Oli e p ocessing consis ed o passing he milled ui s h ough a sie e (po e diame e : 1.5 mm). Du ing his p ocess, pa s o he s ones (seeds) we e emo ed o p oduce “pa ly des oned” OP. In he die s wi h enzyme (ENZ), his p oduc (50 mg/kg) was added o he OP (Na uzyme P50®; Sydney, Aus alia). The nu i ional chemical analysis o p ocessed (OPp) and unp ocessed (OPu) d ied OP was pe o med [41]. Table 3. Chemical composi ion o wo ypes o oli e meal used in he expe imen (as- ed basis). Types o Oli e Meal D ied P ocessed Oli e Pulp (Pa ly Des oned)* O iginal D ied Unp ocessed Oli e Pulp D y ma e (g/kg) 934.50 935.70 Me abolizable ene gy (kcal/kg) 2980.00 1250.00 C ude p o ein (g/kg) 107.30 71.10 C ude ibe (g/kg) 256.00 350.00 Neu al de e gen ibe (α-amylase) (g/kg) 716.00 744.00 Acid de e gen ibe (g/kg) 550.00 584.00 Ash (g/kg) 85.00 62.00 C ude a (g/kg) 130.00 85.00 Calcium (g/kg) 8.20 6.10 Phospho us (g/kg) 0.70 0.60 Soluble suga s (g/kg) 1.70 1.40 S a ch (g/kg) 9.70 10.50 To al polyphenols (g/kg) 3.70 1.90 To al annins (g/kg) 22.90 17.90 * Seeds pa ially emo ed. A he age o 42 days (a e 4 h- as ing o comple e e acua ion o he gu ), one bi d om each eplica e we e andomly selec ed, weighed, and eu hanized (s unned and slaugh e ed). In he slaugh e me hod, s unning by elec ona cosis and subsequen bleeding (by he jugula cou ) we e used. The a e ages o hese bi ds we e calcula ed and used as one expe imen al uni . The expe imen al uni s we e used o measu e ca cass yield and o al cha ac e is ics. Bi ds we e ully de ea he ed ia he d y me hod. Fee we e sepa a ed om he ca cass a he ibio a sal join . Neck, wing ips, gu , and li e we e emo ed, and he ca cass was weighed (cold ca cass weigh , a e chilling). Economically ele an pa s o he ca cass and o al we e sepa a ed. Fi s , b eas muscle (including he skin and s e num) was dissec ed ee om he ca cass. Legs ( highs and d ums icks we e dissec ed by disa icula ion a he hip join and dissec ion o issue om he iliac bone. All abdominal a (including ha a ound he ec um, gizza d, and p o en iculus) was collec ed. Va ious pa s o he ca casses we e dissec ed and weighed sepa a ely. All pa s, including he head, b eas , wings, neck, highs and d ums icks (legs), gizza d, hea , li e , lung, and abdominal a , we e weighed and weigh eco ded. The o al weigh o all dissec ed pa s was ela ed o he whole e isce a ed ca cass. Rela i e pe cen age a ios we e calcula ed acco ding o he ollowing equa ion: 100 x (weigh o componen (s)/e isce a ed ca cass weigh ). Da a analysis o a iance used he wo-way ANOVA p ocedu e. Da a we e submi ed o wo- way analysis o he a iance [42]. The ollowing equa ion was applied: Yijk= μ+Ai+Bj+Ck+ABij+ACik+BCjk+ABCijk+eijkl (1) whe e μ = gene al a e age, Ai = oli e pulp le els, Bj = e ec o p ocessing, Ck = e ec o enzyme supplemen a ion, ABij = e ec o he in e ac ion be ween oli e pulp le els and p ocessing, ACik = e ec o he in e ac ion be ween oli e pulp le els and enzyme supplemen a ion, BCjk = e ec o he in e ac ion be ween oli e pulp p ocessing and enzyme complex in e ac ion e ec , ABCijk = e ec o Ag icul u e 2020, 10, 359 8 o 17 he in e ac ion among oli e pulp le els, oli e pulp p ocessing, and enzyme supplemen a ion, and eijkl = inciden al esidual e ec o obse a ion. A e he s a is ical di e ences we e con i med, he gene al linea model (PROC GLM) was used, and he di e ences be ween means (p ≤ 0.05) we e assessed ia Duncan’s mul iple ange es . 3. Resul s Highe alues o OP did no a ec he ollowing cha ac e is ics: li e body weigh (LW), de- ea he ed body weigh (DW), ull abdomen ca cass weigh (FC), emp y abdomen ca cass weigh (EC), e isce a ed ca cass weigh (EC ), b eas weigh , high and d ums ick weigh (legs), wing weigh , and ela i e b eas and wing weigh s (p >0.05; Table 4). The ela i e b eas weigh was inc eased by OP p ocessing (pa ial des oning) (p>0.05; Table 4). Howe e , a signi ican di e ence, which was no ound in he alues o LW, DW, FC, EC, ela i e EC , b eas , wing, ela i e wing, high and d ums ick, and ela i e high and d ums ick weigh s was due o he OP p ocessing (p >0.05) as shown in Table 4. Ag icul u e 2020, 10, 359 9 o 17 Table 4. Ca cass ai s o b oile s ed die s con aining di e en amoun s o p ocessed (OPp) and unp ocessed (OPu) oli e pulp, wi h and wi hou enzyme (ENZ), in he pe iod 1–6 weeks o age. LW (g) DW (g) FC (g) EC (g) EC (%) B W (g) B W (%) TDW (g) TDW (%) WW (g) WW (%) Die s OPp (50 g/kg) 2770 a 2464 a 2228 a 1743 a 78.3 a 842 a 34.2 a 718 a 29.2 a,b 95 a 3.87 a OPp (50 g/kg) + ENZ 2752 a 2476 a 2295 a 1752 a 76.3 a 857 a 34.5 a 680 a 27.5 b 108 a 4.35 a OPp (100 g/kg) 2807 a 2482 a 2315 a 1763 a 76.2 a 857 a 34.5 a 707 a 28.5 b 110 a 4.40 a OPp (100 g/kg) + ENZ 2782 a 2456 a 2288 a 1738 a 75.9 a 847 a 34.4 a 690 a 28.1 b 106 a 4.33 a OPu (50 g/kg) 2910 a 2564 a 2365 a 1847 a 78.0 a 862 a 33.5 a 845 a 32.8 a 112 a 4.34 a OPu (50 g/kg) + ENZ 2952 a 2572 a 2368 a 1800 a 76.1 a 849 a 33.0 a 749 a 29.2 a,b 111 a 4.32 a OPu (100 g/kg) 2827 a 2490 a 2278 a 1687 a 74.0 a 801 a 32.1 a 673 a 27.0 b 105 a 4.22 a OPu (100 g/kg) + ENZ 3033 a 2635 a 2433 a 1780 a 73.2 a 842 a 32.0 a 735 a 27.9 b 107 a 4.04 a Wi hou OP 2797 a 2447 a 2215 a 1674 a 75.6 a 775 a 31.7 a 672 a 27.5 b 100 a 4.06 a Wi hou OP + ENZ 2872 a 2642 a 2348 a 1820 a 77.5 a 902 a 34.2 a 681 a 25.7 b 109 a 4.16 a S anda d e o s ±114 ±99 ±90 ±70 ±1.4 ±39 ±0.9 ±35 ±0.9 ±6 ±0.20 LW: Li e body weigh ; DW: De ea he ed body weigh ; FC: Full abdomen ca cass weigh ; EC: Emp y abdomen ca cass weigh ; EC : E isce a ed ca cass; B W: B eas weigh ; B W : Rela i e b eas weigh ; TDW: Thigh and d ums ick weigh ; TDW : Rela i e high and d ums ick weigh ; WW: Wing weigh ; WW : Rela i e wing weigh . a,b Means (± s anda d e o s) wi hin each column o die a y ea men s wi h no common supe sc ip di e signi ican ly a p ≤0.05. 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