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Elimination of antibiotic growth promoters from poultry production has encouraged intensive search for relevant alternatives. Prebiotics are proposed as efficient replacements to stimulate colonization/expansion of beneficial microflora in chickens. The aim of this study was to deepen the knowledge on the effect of prebiotic administration on slaughter performance and meat quality traits of broiler chickens by evaluating different routes of their delivery (in ovo vs. in-water vs. in ovo + in-water). At d 12 of incubation, 1,500 eggs (Ross 308) containing viable embryos were randomly allotted into 4 groups and injected in ovo with 0.2 mL solution containing: 3.5 mg/embryo BI (Bi2tos, trans-galactooligosaccharides); 0.88 mg/embryo DN (DiNovo, extract of Laminaria spp.); 1.9 mg/embryo RFO (raffinose family oligosaccharides) and 0.2 mL physiological saline (C). All prebiotics increased final BW compared to C group (P < 0.01), irrespective of delivery route. The prebiotics injected in ovo (T1) or in ovo combined with in-water supplementation (T2) increased carcass weight as compared with in-water group (T3), while T3 had the lowest carcass yield compared to the other groups. All prebiotics increased breast muscle weight and yield (P < 0.01), as well as fiber diameter (P < 0.05). Ultimate meat pH was lower (P < 0.01) in T3 than in T2 group. Meat from chickens treated with prebiotics showed a lower redness index, while lightness and yellowness were not affected by the treatments. Saturated fatty acid (SFA), polyunsaturated fatty acid (PUFA) and n-3 fatty acids contents were higher (P < 0.01), and monounsaturated fatty acid (MUFA) level was lower (P < 0.01) in prebiotic groups compared with C group. Nutritional indexes (n-6/n-3, PUFA/SFA ratio and thrombogenic index) displayed favorable human health-promoting values in the meat of chickens which were treated with prebiotics, irrespective of delivery route. Muscle cholesterol content was not affected by prebiotics. In conclusion, this study has shown that prebiotics can exert positive effects on growth of broiler chickens, carcass and meat quality traits, irrespective of delivery route.