Pluripotency properties of embryonic stem cells isolated from stage X blastoderm of Mazandaran native chicken.

The aim of this study was to isolate Embryonic Stem Cells (ESCs) from native chicken and to characterize their pluripotency properties through the cellular and molecular markers. Samples obtained from fertilized eggs from Mazandaran native hens. Cells were isolated from area of pellucida from stage X native hens' blastoderm. Then the cells were cultured on inactivated mouse SNL feeder cells in the presence of LIF, IGF-1, bFGF, CNTF, OSM, SCF, Il-6, and Il-11 growth factors. The native chickens' ESCs colonies were picked up and subsequently passaged. To characterize the cells, they were analyzed for their alkaline phosphatase activity, and also for the expression of SSEA-4, and TRA-1-60 as embryonic-specific markers at the protein level. Furthermore, the expression of pluripotency (cPouV, Sox2, and Nanog) and cell lineage specific (Cvh, Brachyury, and Gata6) gene markers was evaluated at the level of mRNA using quantitative RT-PCR. Isolated cells were passaged repeatedly and successfully up to ten passages. The stemness of embryonic cells has been approved by the activity of the alkaline phosphatase, presence of the SSEA-4, and TRA-1-60 protein, and expression of the molecular marker (cPouV, Nanog, and Sox-2) genes. The spontaneous differentiation of chicken ESCs confirmed the pluripotecy of the cells in differentiation into specialized cell lineages. Our observation showed that ESCs can be isolated successfully from stage X blastoderm of Mazandaran native chickens and these cells maintain their stemness properties during multi-passages in vitro.

Effects of flavomycin and probiotic supplementation to diets containing different sources of fat on growth performance, intestinal morphology, apparent metabolizable energy, and fat digestibility in broiler chickens.

This study was conducted with broilers to evaluate the effects of growth-promoting antibiotic (flavomycin) and probiotic (7 bacterial species) supplementation in diets containing soybean oil or free fatty acids (FFA) on performance, morphological parameters of the small intestine, apparent digestibility of gross energy (GE) in the ileum, and apparent digestibility of fat in the ileum and total intestinal tract. Eight-hundred and sixty 4-d-old Ross 308 broiler chicks were used in a 3 × 3 factorial arrangement of dietary treatments that comprised 3 additives (without additive, flavomycin, and probiotic) and 3 fat sources (without fat, 30 g/kg of FFA, and 30 g/kg of soybean oil) with 4 pen replicates per treatment. All diets contained chromic oxide (3 g/kg) as an indigestible marker. Body weight and feed intake were recorded weekly over 40 d. Flavomycin interacted positively with soybean oil and FFA causing improvements (P < 0.05) in BW gain. Among the different fat sources, soybean oil significantly increased (P < 0.05) BW gain and jejunal villi height, whereas flavomycin improved (P < 0.05) BW gain and feed conversion when compared with the remaining dietary additives. However, the probiotic negatively affected (P < 0.05) BW gain and feed conversion despite increased (P < 0.05) villi heights of the duodenum, jejunum, and ileum. At 21 and 38 d of age, fat and GE digestibility were higher (P < 0.05) in the ileum and total intestinal tract of birds fed diets containing soybean oil than those of birds fed FFA. Fat and GE digestibility were highest (P < 0.05) among birds fed flavomycin but lowest (P < 0.05) among probiotic-fed birds. Flavomycin addition to soybean oil or FFA diets significantly increased (P < 0.05) fat and GE digestibility when compared with the same diets containing the probiotic. Therefore, soybean oil is a better energy source than FFA, as indicated by increased growth, nutrient digestibility, and jejunal villi height. However, probiotic supplementation to fat-rich diets caused detrimental effects on nutrient digestibility and growth.