Commercially produced chickens have become key food-producing animals in the global food system. The scale of production in industrial settings has changed management systems to a point now very far from traditional methods. During the perinatal period, newly hatched chicks undergo processing, vaccination and transportation, which introduces a gap in access to feed and water. This gap, referred to as the hatching window, dampens the potential for microflora inoculation and as such, prevents proper microbiome, gastrointestinal system and innate immunity development. As a consequence, the industrial production of chickens with a poor microbial profile leads to enteric microbial infestation and infectious disease outbreaks, which became even more prevalent after the withdrawal of antibiotic growth promoters on many world markets (e.g., the EU).This review presents the rationale, methodology and life-long effects of in ovo stimulation of chicken microflora. In ovo stimulation provides efficient embryonic microbiome colonization with commensal microflora during the perinatal period. A carefully selected bioactive formulation (prebiotics, probiotics alone or combined into synbiotics) is delivered into the air cell of the egg on day 12 of egg incubation. The prebiotic penetrates the outer and inner egg membranes and stimulates development on the innate microflora in the embryonic guts. Probiotics are available after the mechanical breakage of the shell membranes by the chick’s beak at the beginning of hatching (day 19). The intestinal microflora after in ovo stimulation is potent enough for competitive exclusion and programs the lifespan condition. We present the effects of different combinations of prebiotic and probiotic delivered in ovo on day 12 of egg incubation on microflora, growth traits, feed efficiency, intestinal morphology, meat microstructure and quality, immune system development, physiological characteristics and the transcriptome of the broiler chickens.We discuss the differences between in ovo stimulation (day 12 of egg incubation) and in ovo feeding (days 17–18 of egg incubation) and speculate about possible future developments in this field. In summary, decades of research on in ovo stimulation and the lifelong effects support this method as efficient programming of lifespan conditions in commercially raised chickens.

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.

The purpose of this study was to examine the effect of in ovo injection of 2 different prebiotics, DiNovo (DN; Laminaria spp., extract containing laminarin and fucoidan) and Bitos (BI; non-digestive trans-galactooligosaccharides from milk lactose digested with Bifidobacterium bifidum NCIMB 41171), on growth, slaughter traits, intramuscular fat percentage (IF) and muscle fiber diameter, and lipid oxidation of meat in chickens reared under commercial conditions, following an in ovo trial protocol. On d 12 of embryonic incubation, 350,560 Ross 308 eggs were randomly divided into 3 experimental groups and automatically injected in ovo with: physiological saline (control group), BI at dose of 3.5 mg/embryo and DN at dose of 0.88 mg/embryo. Hatched chicks (males and females) were allocated dependent on treatment group into 3 poultry houses on each farm (3 farms in total) with a stocking density of 21.2 to 21.5 chicks/m At 42 d of age, 14 randomly chosen birds (7 males and 7 females), per each treatment from each farm, were individually weighed and slaughtered. The results showed no significant differences of final number of chickens/chicken house, mortality, BW per treatment, stocking density (kg/m), feed intake, feed conversion rate (FCR), and European Broiler Index among 3 experimental groups. Treatments with BI and DN were associated with slight increases (P > 0.05) in average BW and a minor improvement (P > 0.05) of FCR in BI group. Slaughtered chickens from DN and BI treated groups had significantly increase of BW, carcass weight, carcass yield, and breast muscle weight compared with the control group. IF and muscle fiber diameter were similar among groups. Males had significantly higher slaughter traits compared to females, except for breast muscle yield. The prebiotic treatments led to a higher lipid oxidation in meat, even if the detected TBA reactive substances were below the critical value recognized for meat acceptability. In conclusion, in ovo administration of prebiotics was associated with improvements in a number of parameters of relevance to commercial poultry production.