Background: Germ cells are critical for any species that multiplies through sexual reproduction. Results: We found 173 candidate key genes and 18 key signaling pathways that are differentially activated. Conclusion: Our results showed the crucial genes and pathways involved in the regulation of chicken male germ cell differentiation. Significance: This study narrows the range of functional genes and pathways during ESC differentiation.

BackgroundNLRC5 is a member of the CARD domain containing, nucleotide-binding oligomerization (NOD)-like receptor (NLR) family, which recognizes pathogen-associated molecular patterns (PAMPs) and initiates an innate immune response leading to inflammation and/or cell death. However, the specific role of NLRC5 as a modulator of the inflammatory immune response remains controversial. It has been reported to be a mediator of type I IFNs, NF-kB, and MHC class I gene. But no study on NLRC5 function has been reported to date in chickens. In the current study, we investigated the role of NLRC5 in the regulation of IFNA, IFNB, IL-6, and MHC class I in the chicken HD11 macrophage cell line, by using RNAi technology. HD11 cells were transfected with one of five siRNAs (s1, s2, s3, negative-siRNA, or a mixture of s1, s2, s3-siRNAs). After 24 hours, cells were exposed to LPS or poly (I:C) or a vehicle control. Gene expression of NLRC5, IFNA, IFNB, IL-6, and MHC class I at 2, 4, 6, and 8 hours post stimulation (hps) was quantified by qPCR.ResultsThe expression of NLRC5, IFNA, IFNB, and IL-6 genes in negative irrelevant transfection controls was up-regulated at 2 hps after LPS treatment compared to the vehicle controls. S3-siRNA effectively knocked down NLRC5 expression at 4 hps, and the expression of IFNA and IFNB (but not IL-6 and MHC class I) was also down-regulated at 4 hps in s3-siRNA transfected cells, compared to negative irrelevant transfection controls. Stimulation by LPS appeared to relatively restore the decrease in NLRC5, IFNA, and IFNB expression, but the difference is not significant.ConclusionsFunctional characterization of chicken NLRC5 in an in vitro system demonstrated its importance in regulating intracellular molecules involved in inflammatory response. The knockdown of NLRC5 expression negatively mediates gene expression of IFNA and IFNB in the chicken HD11 cell line; therefore, NLRC5 likely has a role in positive regulation of IFNA and IFNB expression. No direct relationship was found between NLRC5 knockdown and IL-6 and MHC class I expression. Future studies will further clarify the roles of NLRC5 and other NLRs in infectious diseases of chickens and may increase the efficacy of antiviral vaccine design.

The present study was conducted to evaluate the effects of different rearing methods and stocking densities on carcass yield and proximate composition of meat in small-sized meat ducks. A total of 555 one-day-old birds were randomly allocated to six treatment groups (three replicates per treatment, sex ratio 1/1) with a 2 × 3 factorial arrangement of two rearing methods (reared in cage or net) and three stocking densities (5 [low], 7 [medium], or 9 [high] birds/m 2 ) until day 70. Five male and five female birds from each replicate were randomly selected and processed to determine the carcass yield. Proximate composition was determined by proximate analysis using the breast and thigh muscles. There was no interaction effect between the rearing method and stocking density on carcass yield. The rearing method affected the thigh muscle rate, which was higher in the cage groups ( P < 0.05). The final BW and abdominal fat rate decreased with increasing density ( P < 0.05), whereas the thigh muscle rate increased ( P < 0.05). There were significant interaction effects ( P < 0.05) between the rearing method and stocking density on the content of protein, fat, and collagen. The content of fat and moisture was greater and lower, respectively, in the cage groups ( P < 0.05). The content of moisture, fat, and collagen with a medium density was higher ( P < 0.05). In addition, the content of protein and fat was lower in the ducks fed in nets at low and high densities ( P < 0.05), respectively; the collagen content of breast and thigh muscle was lower in the ducks fed in cages and nets, respectively, at a low density ( P < 0.05). Our findings provide valuable insights into the single and interactive effects of the rearing method and stocking density on duck slaughter performance and proximate composition of meat. The results indicate that a rearing system with a cage pattern and a medium density is better than other arrangements for small-sized meat ducks.

Avian leukosis virus subgroup J (ALV-J) can induce myeloid tumors and hemangiomas in chickens and causes severe economic losses with commercial layer chickens and meat-type chickens. Here, we generated ribominus RNA sequencing data from three normal chicken spleen tissues and three ALV-J-infected chicken spleen tissues. Structure analysis of transcripts showed that, compared to mRNAs and lncRNAs, chicken circRNAs shared relatively shorter transcripts and similar GC content. Differentially expression analysis showed 152 differentially expressed circRNAs with 106 circRNAs up regulated and 46 circRNAs down regulated. Through comparing differentially expressed circRNA host genes and mRNAs and performed ceRNA network analysis, we found several tumor or immune-related genes, in which, there were four genes existed in both differentially expressed mRNAs and circRNA host genes (Dock4, Fmr1, Zfhx3, Ralb) and two genes (Mll, Aoc3) involved in ceRNA network. We further characterized one exon-intron circRNA derived from HRH4 gene in the ceRNA network, termed circHRH4, which is an abundant and stable circRNA expressed in various tissues and cells in chicken and localizes in cytoplasm. Our results provide new insight into the pathology of ALV-J infection and circRNAs may also mediate tumorigenesis in chicken.

BackgroundGut microbial composition is dependent on diet. Geese are herbivores and can digest crude fibre, but the relationship between composition of the microbiota and a fibre-rich diet in geese is not well understood.ResultsHere, caecal and faecal samples were collected simultaneously from all-grass-fed geese and high-grain-fed geese and the hypervariable V3–V4 regions of the bacterial 16S rRNA gene were sequenced. The results was identified that high-grass-fed geese possessed significantly higher alpha diversity both in caecum and faeces compared with that in all-grain-fed geese. In addition, the composition of dominant bacterium occurred remarkable shifting due to different diet patterns, Firmicutes were more abundant in all-grass-fed geese, whereas Bacteroidetes were abundant in high-grain-fed geese. Fusobacteria and Deferribacteres were obviously present in high-grain-fed geese and few in all-grass-fed geese. Most importantly, some specific microorgnisms such as Ruminococcaceae, Lachnospiraceae and Bacteroidaceae which may associated with cellulose-degrading that were characterized to show distinctly diverse between the two diet patterns. PICRUSt analysis revealed the metabolic pathways such as carbohydrate and amino acid metabolism were overrepresented in all-grass-fed geese.ConclusionsIn conclusion, Firmicutes and Bacteroidetes were identified abundantly when the geese was fed with all-grass feed and high-grain feed, respectively. And Ruminococcaceae, Lachnospiraceae and Bacteroidaceae were recognized as main cellulose-degrading bacteria in the geese. The functional profiles of gut microbiota revealed the dominant microbiota communities were involved mainly in the carbohydrate metabolism in all-grass-fed geese.

MicroRNA (miRNA)-1 and miRNA-133 are derived from the same bicistronic pairs with roles in skeletal muscle development. Many investigations have focused on the role of miRNA-1 and miRNA-133 in the regulation of skeletal muscle development in mammals and fish. However, the mechanisms of miRNA-1 and miRNA-133 underlying the differences in skeletal muscle development between different breeds are not well known.Our study found that the weights of body and breast at 42 days of age were greater in Cherry Valley ducks than in Putian ducks and the areas of breast muscle fibers increased with age; the areas of muscle fibers of Cherry Valley ducks were always greater than those of Putian ducks. Besides, quantitative reverse-transcriptase polymerase chain reaction analysis revealed that relatively high levels of miRNA-1 and miRNA-133 were detected in heart, breast, and leg muscles compared with the liver, spleen, lung, kidney, and the expression levels of miRNA-1 and miRNA-133 remained stable in the embryo stage, and in the growth period, the fluctuation in miRNA expression levels in Putian ducks was considerably higher than that in Cherry Valley ducks, especially from 7 to 28 days.However, in the late growth period, the expression of miRNA-1 and miRNA-133 of CherryValley duck was higher than that of Putian duck, which may indicate that miRNA-1 and miRNA-133 play a more important role during the growth period. To determine the function of miRNA-1 and miRNA-133 in skeletal muscle development, we found that the overexpression of miRNA-1, but not miRNA-133, promoted fusion of adjacent myoblasts.By contrast, a repressor of miRNA-1 promoted, whereas a miRNA-133 inhibitor inhibited, myoblast proliferation. Accordingly, the expression levels of myocyte enhancer factor 2D (MEF2D) and myogenic differentiation (MYOD) were significantly increased by an miRNA-1 mimic and the miRNA-133 inhibitor. In addition, we found that the expression levels of miRNA-1 significantly affected the expression of histone deacetylase 4 (HDAC4), and miRNA-133 affected serum response factor (SRF) and transforming growth factor β receptor 1 (TGFBR1) levels. However, dual-luciferase reporter assays revealed that only miRNA-1 directly inhibited pGL-HDAC4 luciferase reporter activity, whereas miRNA-133 did not affect pGL-SRF or pGL-TGFBR1 fluorescence activity. Taken together, these results suggest that miRNA-1 targets HDAC4 to promote the differentiation of duck myoblasts and miRNA-133 may affect SRF and TGFBR1 expression to promote proliferation, which indicates that miRNA-1 and miRNA-133 play different important roles in skeletal muscle development. K E Y W O R D S duck, microRNA-1, microRNA-133, proliferation and differentiation, skeletal muscle J Cell Physiol. 2019;234:3490-3499. wileyonlinelibrary.com/journal/jcp 3490 |

Adipocytes are derived from pluripotent mesenchymal stem cells through adipogenesis. Pre-adipocyte differentiation in poultry greatly influences fat deposition and meat quality. Circular RNAs (circRNAs) have an important function in cancer and some differentiation processes. Herein, high-throughput transcriptome sequencing was used to detect circRNAs present in cherry valley duck pre-adipocyte and adipocyte differentiation over 3 days. We identified 9,311 circRNAs and 141 differentially expressed circRNAs. Sequencing results were verified through qRT-PCR using seven randomly selected circRNAs, and competing endogenous RNA (ceRNA) networks were exhibited by ten important circRNAs in duck adipocyte differentiation. circRNA plexin A1 (circ-PLXNA1) was detected in duck adipocytes and mainly expressed in adipose, leg muscle and liver. Inhibition of circ-PLXNA1 limited the differentiation of duck adipocyte. There were four corresponding miRNAs for circ-PLXNA1 and 313 target genes for those miRNAs. CeRNA“circ-PLXNA1/miR-214/CTNNB1 axis” was focused and verified by a dual-luciferase reporter experiment. After co-transfection of cells with si-circ-PLXNA1 and miR-214 mimics, the expression level of CTNNB1 was down-regulated, triglyceride content and the adipogenic capacity of preadipocytes decreased. While there were no significant change after si-CTNNB1 transfection. All these results provide further insight into the circRNAs, especially for circ-PLXNA1 in duck adipocyte differentiation.

This study was performed to evaluate the effects of different rearing methods on the growth performance, carcass yield, and meat quality of small-sized meat ducks. A total of 420 healthy 21-day-old birds was randomly allocated to 2 treatment groups (6 replicates per treatment, sex ratio 1/1) and subjected to 2 rearing methods (furnished cage and plastic wire-floor) until d 63. Growth performance was measured in all birds. Three males and 3 females from each replicate were randomly selected and evaluated to determine the carcass yield and meat quality. In terms of growth performance, the rearing method affected the final body weight, average daily feed intake, and average daily gain, which were higher in the cage group ( P < 0.05) than in the floor group, with a similar feed/gain in both groups. For slaughter performance, ducks in the cage group showed a higher abdominal fat yield and lower gizzard yield than those in the floor group ( P < 0.05). For meat quality, the L* value of the breast muscle was higher in the cage group than in the floor group ( P < 0.05). The pH recorded at 1 h was lower and pH recorded at 24 h was higher in the cage group ( P < 0.05). The shear force and water loss rate were both lower in the cage group ( P < 0.05). Additionally, the moisture content was lower and intramuscular fat content was higher in ducks fed in cages ( P < 0.05). Our results indicate that the cage rearing system improved the growth performance and meat quality of ducks, which is appropriate for small-sized meat ducks.