Weimin Luan

In this study, the effects of synbiotic inclusion at the intra-amniotic stage in layer chicks were evaluated with different parameters, such as performance, immunological function, intestinal development, and cecal microflora content. A total of 1,200 eggs with fertile embryos were allocated into four treatment groups. For every treatment, five replicates were used, and 60 eggs were included in each replicate. The following four treatment groups were established: the non-injected group, 0.9% physiological saline injection (saline) group, 1 × 10⁶ CFU/egg Lactobacillus plantarum injection (probiotic) group, and 1 × 10⁶ CFU/egg L. plantarum + 2 mg/egg Astragalus polysaccharide injection (synbiotic) group. In ovo injection was carried out at 18.5 days of incubation. The results showed that in ovo injection of probiotics or synbiotics did not affect the hatching or growth performance of the chicks but significantly increased their feed intake (FI), body weight (BW), and the feed conversion ratio (FCR). Additionally, in ovo injection of synbiotics enhanced the levels of serum interleukin-2 (IL-2), interferon-γ (IFN-γ), and secretory immunoglobulin A (SIgA) in intestinal lavage fluid and the histomorphological development of the small intestine. Our results also indicated that intra-amniotic synbiotic injection significantly increased Lactobacillus and Bifidobacterium colonization while decreasing the relative abundance of Escherichia coli in the chicken cecum (P < 0.05). In summary, in ovo injection of synbiotics had positive impacts on the performance, immunological function, gut development, and microbiota of growing chicks.

In this study, immunogenic efficacies of in ovo administration of Astragalus polysaccharide (APS) along with live Newcastle disease vaccine (live ND vaccine) (live VG/GA strain) were evaluated. Four hundred fertilized eggs were randomly divided into four groups (n = 100/group), and vaccinated in ovo, respectively, with solutions of APS, live ND vaccine, live ND vaccine combined with APS, and 0.9% physiological saline into their amniotic fluid on d 18.5 of incubation. Significant improvement of chicks’ development was displayed in those vaccinated with live ND vaccine adjuvanted with APS in ovo, manifested as enhanced hatchability and gaining weight. Moreover, in ovo administration of live NDV vaccine plus APS could significantly enhance the serum anti‐NDV antibody titres and interferon gamma (IFN‐γ), interleukin (IL)‐2, IL‐4 and IL‐6 concentrations, promote lymphocyte proliferative capability as well as improve the frequencies of CD4⁺ and CD8⁺ T cells in peripheral blood. Overall results indicated in ovo administration of live ND vaccine adjuvanted with APS could stimulate stronger humoral and cellular responses in newly hatched chicks.

Summary: TRIM28/KAP1/TIF1β was identified as a universal transcriptional co-repressor and is critical for regulating post-fertilization methylation reprogramming in preimplantation embryos. In this study, three siRNAs (si647, si742, and si1153) were designed to target the TRIM28 mRNA sequence. After transfection of the mixture of the three siRNA (siMix) into bovine fibroblast cells, the most effective one for TRIM28 knockdown was selected. By injecting RNAi directed against TRIM28 mRNA, we found that TRIM28 knockdown in oocytes had the most effect on the H19 gene, in which differentially methylated region (DMR) methylation was almost completely absent at the 2-cell stage (1.4%), while control embryos showed 74% methylation. In addition, global H3K9me3 levels at the 2-cell stage were significantly higher in the in vitro fertilization (IVF) group than in the TRIM28 knockdown group (PPTrim28 transcripts are lost during SCNT. TRIM28 is required for the maintenance of methylation imprints in bovine preimplantation embryos, and the loss of TRIM28 during SCNT may contribute to the unfaithful maintenance of imprints in cloned embryos.

Major royal jelly protein 1 (MRJP1, the most abundant protein of honeybee royal jelly) has been recognized as a valuable functional foods and therapeutic protein for human beings. There is great potential using transgenic technology to express recombinant MRJP1 in milk in order to improve the human health conditions. For expression of MRJP1 in milk, mammary gland-specific expression vector (pBC1-MRJP1-NEOr) carrying the expression cassette composed of β-casein promoter, cDNA encoding MRJP1 (with bovine β-casein signal peptide) and a Neor selection marker was constructed, and expressed in human MCF-7 cells. To examine the expression of MRJP1, a His-tag protein cDNA fragments were fused on the 5' end of MRJP1 gene using PCR amplification. This vector was shown to be expressed successfully, thus potentially enabling the generation of transgenic cows to be used as mammary gland bioreactors.