Orf is a highly contagious zoonotic disease of small ruminants caused by Parapoxvirus. Kisspeptin, encoded by the KISS1 gene with its cognate receptor GPR-54 is recognized as an upstream orchestrator in the hypothalamic-pituitary-gonadal axis. This study was designed to construct a DNA vaccine that produces a fused peptide composed of a major immunodominant protein of the orf virus (B2L) and kisspeptin-54, a neuropeptide with recognized roles in mammalian reproductive biology. The administration of this recombinant vaccine is shown to produce a significant antibody and cell-mediated immune response directed against B2L compared to the control group (p < 0.05). Furthermore, we found that rats inoculated with PBK-asd vaccine up-regulated antigen-mediated splenocyte proliferation and significantly raised antigen-specific tumor necrosis factor-alpha (TNFα-), interferon-gamma (IFN-ϒ) and interleukin (IL-2) compared to the control group (p < 0.05). This recombinant vaccine also stimulated antibody responses to kisspeptin and decreased serum luteinizing hormone and testosterone levels. Moreover, the current recombinant vaccine caused testicular atrophy and arrested spermatogenesis. It is concluded that this recombinant B2L and Kisspeptin-54 vaccine could be a promising approach for construction of bivalent orf virus and immunocastration vaccine. Furthermore, we concluded that the orf virus envelope protein (B2L) could be used as an immunomodulator for kisspeptin-54 to produce a strong antibody response.

Background The microscopic process of postnatal testicular development in early-puberty animals is poorly understood. Therefore, in this study, 21 male Yiling goats with average ages of 0, 30, 60, 90, 120, 150 and 180 days old (each age group comprised three goats) were used to study the changes in organs, tissues and transcriptomes during postnatal testicle development to obtain a broad and deep insight into the dynamic process of testicular transition from growth to spermatogenesis in early-puberty animals. Results The inflection point of testicular weight was at 119 days postpartum (dpp), and the testicular weight increased rapidly from 119 dpp to 150 dpp. Spermatozoa were observed in the testis at 90 dpp by using haematoxylin–eosin staining. We found from the transcriptome analysis of testes that the testicular development of Yiling goat from birth to 180 dpp experienced three stages, namely, growth, transition and spermatogenesis stages. The goats in the testicular growth stage (0–60 dpp) showed a high expression of growth-related genes in neurogenesis, angiogenesis and cell junction, and a low expression of spermatogenesis-related genes. The goats aged 60–120 dpp were in the transitional stage which had a gradually decreased growth-related gene transcription levels and increased spermatogenesis-related gene transcription levels. The goats aged 120–180 dpp were in the spermatogenesis stage. At this stage, highly expressed spermatogenesis-related genes, downregulated testicular growth- and immune-related genes and a shift in the focus of testicular development into spermatogenesis were observed. Additionally, we found several novel hub genes, which may play key roles in spermatogenesis, androgen synthesis and secretion, angiogenesis, cell junction and neurogenesis. Moreover, the results of this study were compared with previous studies on goat or other species, and some gene expression patterns shared in early-puberty mammals were discovered. Conclusions The postnatal development of the testis undergoes a process of transition from organ growth to spermatogenesis. During this process, spermatogenesis-related genes are upregulated, whereas neurogenesis-, angiogenesis-, cell junction-, muscle- and immune-related genes are downregulated. In conclusion, the multipathway synergy promotes testicular transition from growth to spermatogenesis in early-puberty goats and may be a common rule shared by mammals.

Simple Summary: Zinc and Copper are two important trace minerals, which are involved in numerous vital biological activities in animal's body, such as enzyme activation and enhancement of immune function, growth, reproduction, DNA synthesis, cell division, and neurotransmitters production. Recently, the application of trace mineral organic complexes in animal feed received much more attention than the inorganic sources. The organic sources can contribute significantly to improving animals' health and reproduction, as the minerals are more biologically available and absorbable than they are when coming from the inorganic sources. In this study, three injectable varieties of Zn and Cu supplementation, sulfate, inorganic (loaded with montmorillonite), and novel organic (ligand with triazine hydrazone), were tested with weaned rabbits. The results revealed that these three mineral types vary at the most biological responses, and only one category of our novel organic complexes provided consistent animal performance improvement, including weight gain, serum antioxidant, meat quality, intestine morphometry, and the expression of peptide growth factors and cytokine genes. To our knowledge, this is the first work on the Zn and Cu with triazine hydrazone ligand as two organic complexes in rabbits.Abstract: Two novel transitional organic Zn/Cu complexes based on a new biocompatible bidentate triazine-hydrazone ligand (Thz) was designed, synthesized, and evaluated in this study. This study evaluated the effects of injecting 60 mg of Zn and 40 mg of Cu in three different forms, twice per week, for eight weeks on growth performance, expression of growth factors and cytokine genes, carcass yield, blood biochemicals, and intestinal morphology in weaned rabbits. The tested complexes were sulfate (Cu/ZnSO 4 ), montmorillonite (Cu/Zn-Mnt), and triazine hydrazone (Cu/Zn-Thz). A total of 60 Animals 2019, 9, 1134 2 of 17 V-line weaned rabbits at four weeks of age were assigned to four treatments (n = 15), which were intramuscularly injected with 0.5 mL of either (1) saline (control) or saline containing (2) Cu/ZnSO 4 , (3) Cu/Zn-Mnt, or (4) Cu/Zn-Thz. Compared to the controls, the rabbits injected with Cu/Zn-Thz showed a higher (p < 0.01) growth rate, carcass yield (p < 0.05), and liver expression of insulin like growth factor-1 (IGF-1), growth hormone receptor (GHR), fibroblast growth factor-1 (FGF1), and transforming growth factor beta-1 (TGFB1) (p < 0.05), as well as better jejunum morphometric variables (p < 0.05). On the other hand, mRNA of FGF1, TGF1, TCIRG1, and adenosine deaminase (ADA) were higher expressed (p < 0.05) in the spleen tissues of Cu/Zn-Mnt group. Collectively, the results indicated that our novel synthesized organic complexes of Zn/Cu-Thz proved to be a suitable feed supplement, as it increased rabbit productive performance through enhancing expression of peptide growth factors and cytokine genes.

No abstract

The melanocortin 4 receptor (MC4R) gene is expressed in the appetite-regulating areas of the brain and is engaged in the leptin signaling pathway. Although previous studies have identified variants in the coding region of the sheep MC4R gene showing significant associations with birth weight, weaning weight, and backfat thickness, no such associations have been reported for the promoter region. Besides, the essential promoter region of the sheep MC4R has not been delineated. In this study, to better understand the transcriptional regulation of MC4R and to elucidate the association between regulatory variants and haplotypes with body measurement traits in sheep, we cloned and characterized the MC4R promoter. We found that the minimal promoter of the gene is located within the region −1207/−880 bp upstream of the first exon. Real-time quantitative PCR (RT-qPCR) data revealed the mRNA expression of the MC4R gene had a significant difference between sex and age. In the association analysis, eight single nucleotide polymorphisms (SNPs) had a significant association with one or more traits (p < 0.05); of these, two SNPs were novel. Notably, individuals with haplotype H1H2 (CT-GA-GT-GA-GT-GA-GA-CG) were heavier in body weight than other haplotypes. Altogether, variations in the MC4R gene promoter, most notably haplotype H1H2, may greatly benefit marker-assisted selection in sheep.