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Owing to its high similarity to human hepatitis B virus (HBV), duck HBV (DHBV) is often used as an essential model for HBV research. Although intergenotypic recombination of HBV is common, it remains unclear whether the intergenotypic recombination of human HBV is exactly the same as that of DHBV. In this study, 119 serum samples of duck and goose were collected from 51 farms (29 duck and 22 goose farms) in the central and eastern regions of China. A total of 22 strains isolated from the 22 DHBV positive flock were sequenced. Genome sequence alignment revealed that the duck- and goose-origin strains shared the highest and lowest similarities (99.7 and 90.52%, respectively). The complete genomes of these DHBV and 31 reference strains were analyzed using phylogenetic methods and classified into 3 clusters, which corresponded to the previously identified DHBV-I, DHBV-II, and DHBV-III branches. Recombination analyses of the 53 DHBV genomes indicated 2 major intergenotypic recombination events with high confidence values. These recombination events occurred between the genotypes of the Chinese isolates Y180813HB (Chinese branch [DHBV-Ⅰ]) and E170101AH (Chinese branch [DHBV-Ⅱ]) and the Western isolate DHBV-XY (Western branch [DHBV-Ⅲ]), resulting in the emergence of 2 Chinese recombinant isolates Y190303HN and Y170101HB. In addition, 40% (2/5) goose-origin and 58.8% (10/17) duck-origin DHBV in this study harbored the mutation site of G133E in preS, which promote the pathogenicity of DHBV. This is the first study to report on the genome analysis and recombination characterization of DHBV isolated from Chinese geese. Further, continuous investigation and molecular identification of DHBV should be conducted to attract researchers' attention., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

The aim of the study was to analyze some slaughter traits in 3 breeds of geese: the commercial hybrids of White Kołuda (W-31), and birds of conservative flocks of Pomeranian (Po) and Kielecka (Ki) geese depending on sex and length of rearing period, and to determine the relationships between the analyzed traits and factors. A total of 19 traits divided into 2 groups-measured and calculated traits-were statistically analyzed. The measured traits (g) included 11 parameters: preslaughter weight, carcass weight, weights of breast muscle, thigh and drumstick, abdominal fat, skin with subcutanenous fat, neck without skin, skeleton with dorsal muscles, wings with skin, total muscles as the sum of all breast and leg muscles, and the trait broth elements as the total weight of neck, skin, skeleton and wings. The calculated traits included 8 parameters (%): dressing percentage-carcass weight to preslaughter weight, meatiness-total breast and leg muscle weight to carcass weight, abdominal fat weight to carcass weight, weight of skin with subcutaneous fat to carcass weight, weight of neck without skin to carcass weight, weight of skeleton with dorsal muscles to carcass weight, weight of wings with skin to carcass weight, as well as total weight of neck, skin, skeleton, and wings. The presented results concerning the analysis of selected slaughter traits of the Kielecka, Pomeranian and White Kołuda geese indicate that these birds exhibit good slaughter value (dressing percentage from 60.80 to 66.50%). The selected values of this parameter were influenced mainly by genotype and to a lesser extent by sex. The White Kołuda geese were characterized by significantly higher values of the majority of the analyzed slaughter traits, both measured and calculated ones. In turn, the lighter domestic geese of regional breeds were characterized by significantly higher carcass meat content (from 31.69 to 35.13 % vs. from 29.28 to 31.80 %) and lower carcass fatness (abdominal fat and skin with subcutaneous fat from 21.26 to 25.45 % vs. from 30.81 to 33.14 %). This points to the possibility of using these breeds of geese in breeding work aimed to produce a hybrid characterized by medium body weight (intermediate between these values for the White Kołuda geese and Kielecka or Pomeranian geese), good dressing percentage as well as high carcass meat content and low carcass fatness., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Nutrition and energy levels have an important impact on animal growth, production performance, disease occurrence and health recovery. Previous studies indicate that melanocortin 5 receptor (MC5R) is mainly involved in the regulations of exocrine gland function, lipid metabolism and immune response in animals. However, it is not clear how MC5R participates in the nutrition and energy metabolism of animals. To address this, the widely used animal models, including the overfeeding model and the fasting/refeeding model, could provide an effective tool. In this study, the expression of MC5R in goose liver was first determined in these models. Goose primary hepatocytes were then treated with nutrition/energy metabolism-related factors (glucose, oleic acid and thyroxine), which is followed by determination of MC5R gene expression. Moreover, MC5R was overexpressed in goose primary hepatocytes, followed by identification of differentially expressed genes (DEGs) and pathways subjected to MC5R regulation by transcriptome analysis. At last, some of the genes potentially regulated by MC5R were also identified in the in vivo and in vitro models, and were used to predict possible regulatory networks with PPI (protein-protein interaction networks) program. The data showed that both overfeeding and refeeding inhibited the expression of MC5R in goose liver, while fasting induced the expression of MC5R . Glucose and oleic acid could induce the expression of MC5R in goose primary hepatocytes, whereas thyroxine could inhibit it. The overexpression of MC5R significantly affected the expression of 1381 genes, and the pathways enriched with the DEGs mainly include oxidative phosphorylation, focal adhesion, ECM-receptor interaction, glutathione metabolism and MAPK signaling pathway. Interestingly, some pathways are related to glycolipid metabolism, including oxidative phosphorylation, pyruvate metabolism, citrate cycle, etc. Using the in vivo and in vitro models, it was demonstrated that the expression of some DEGs, including ACSL1 , PSPH , HMGCS1 , CPT1A , PACSIN2 , IGFBP3 , NMRK1 , GYS2 , ECI2 , NDRG1 , CDK9 , FBXO25 , SLC25A25 , USP25 and AHCY , was associated with the expression of MC5R , suggesting these genes may mediate the biological role of MC5R in these models. In addition, PPI analysis suggests that the selected downstream genes, including GYS2 , ECI2 , PSPH , CPT1A , ACSL1 , HMGCS1 , USP25 and NDRG1 , participate in the protein-protein interaction network regulated by MC5R . In conclusion, MC5R may mediate the biological effects caused by changes in nutrition and energy levels in goose hepatocytes through multiple pathways, including glycolipid-metabolism-related pathways.

Goose is an important poultry commonly raised for meat. The early growth performance of geese significantly influences their market weight and slaughter weight, affecting the poultry industry's economic benefits. To identify the growth surge between the Shitou goose and the Wuzong goose, we collected the early growth body traits from 0 to 12 weeks. In addition, we investigated the transcriptomic changes in leg muscles at the high growth speed period to reveal the difference between the two geese breeds. We also estimated the growth curve parameters under three models, including the logistic, von Bertalanffy, and Gompertz models. The results showed that except for body length and keel length, the best-fitting model between the body weight and body size of the Shitou and Wuzong was the logistic model. The growth turning points of Shitou and Wuzong were 5.954 and 4.944 weeks, respectively, and the turning point of their body weight was 1459.01 g and 478.54 g, respectively. Growth surge occurred at 2-9 weeks in Shitou goose and at 1-7 weeks in Wuzong goose. The body size traits of the Shitou goose and Wuzong goose showed a trend of rapid growth in the early stage and slow growth in the later stage, and the Shitou goose growth was higher than the Wuzong goose. For transcriptome sequencing, a total of 87 differentially expressed genes (DEGs) were identified with a fold change ≥ 2 and a false discovery rate < 0.05. Many DEGs have a potential function for growth, such as CXCL12 , SSTR4 , FABP5 , SLC2A1 , MYLK4 , and EIF4E3 . KEGG pathway analysis identified that some DEGs were significantly enriched in the calcium signaling pathway, which may promote muscle growth. The gene-gene interaction network of DEGs was mainly related to the transmission of cell signals and substances, hematological system development, and functions. This study can provide theoretical guidance for the production and breeding management of the Shitou goose and Wuzong goose and help reveal the genetic mechanisms underlying diverse body sizes between two goose breeds.

Changle goose in Fujian, China is a rare genetic resource and in urgent need to be protected. Understanding the characteristics of digestive physiology and spatial variation of gastrointestinal microbiota is crucial for developing nutritional intervention strategies to improve intestinal health and production performance of goose. Hence, histomorphological assay was used for observing development status of proventriculus, jejunum, and cecum in 70-day-old Changle geese, whereas digesta from 6 alimentary canal locations (crop, proventriculus, gizzard, jejunum, cecum, and rectum) were collected for 16S rRNA gene sequencing and short chain fatty acids (SCFAs) quantitative analysis. The histomorphological observation indicated that the jejunum and cecum of Changle goose were well developed. The alpha diversity analysis revealed that, except rectum, microbiota in other noncecum sections were in high diversity as cecum. The Nonmetric MultiDimensional Scaling (NMDS) analysis showed that microbial community of proventriculus, gizzard, and jejunum formed a cluster, which distinctly discrete with the microbiota of the other gastrointestinal locations. Additionally, the proportions of Proteobacteria, Bacteroidota, and Campilobacterota at the phylum level and Lactobacillus, Streptococcus, Helicobacter, and Subdoligranulum at the genus level exhibited tremendous alternations among different gastrointestinal locations. The characteristic bacterial composition in each section was further disclosed by analyzing the core and feature Amplicon Sequence Variants (ASVs) and SCFAs pattern. Importantly, 7 body-weight-associated ASVs and 2 cecum-development-related ASVs were identified via correlation analysis. In a whole, our findings provided the first insights into the specialized digestive physiology of Changle geese and distinctive regional distribution of gastrointestinal microbiota, which laid the important foundation for improving growth performance through microbiota manipulation in geese., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Egg production is an important economic trait in the Chinese goose industry. Due to the low heritability of annual egg production traits in geese, large-scale individual selection based on annual egg production measurements cannot be carried out. Therefore, new selection methods must be applied for large-scale early selections. To screen for effective molecular markers for early Yangzhou geese selection, the genotypes and gene frequencies of mutated loci of five candidate genes related to egg production, MAGI-1, ACSF2, ASTN2, KIAA1462, and ARHGAP21, were detected and analyzed by PCR-direct sequencing.Furthermore, correlation analysis was performed with annual egg mass and body weight at the point of lay and egg weight, and the results were as follows:Magi-1 (Record-106975)was A > G, ACSF2 (Record-106582)was A > C, ASTN2 (Record-111407)was A > T, KIAA1462 (Record-134172)was A > T, and the base of ARHGAP21 (Record-112359) was G > T. At all the five loci above, the Yangzhou geese population followed the Hardy-Weinberg equilibrium (P > 0.05). The results of the association analysis between different genotypes and production performance showed no significant differences in annual egg production, body weight at the point of lay, and egg weight, among different genotypes (P > 0.05) at the mutation loci of MAGI-1 and ASTN2. At the ACSF2 and KIAA1462, the annual egg production of AC was significantly higher than that of AA and CC (P < 0.05), the annual egg production of TT was significantly higher than that of AA (P < 0.05), and there were no significant differences in body weight at the point of lay and egg weight, among the three genotypes (P > 0.05). At ARHGAP21, the body weight at the lay point of the TT genotype was the highest, which was significantly higher than that of GG (P < 0.05); however, there was no significant difference with the heterozygous GT genotype for this trait (P > 0.05). Therefore, Genotype AC at ACSF2 and genotype TT at KIAA1462 could be used as favorable genotypes for egg production, and genotype TT at ARHGAP21 could be used as a favorable genotype for weight in Yangzhou geese., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Skeletal muscle development from embryonic stages to hatching is critical for poultry muscle growth, during which DNA methylation plays a vital role. However, it is not yet clear how DNA methylation affects early embryonic muscle development between goose breeds of different body size. In this study, whole genome bisulfite sequencing (WGBS) was conducted on leg muscle tissue from Wuzong (WZE) and Shitou (STE) geese on embryonic day 15 (E15), E23, and post-hatch day 1. It was found that at E23, the embryonic leg muscle development of STE was more intense than that of WZE. A negative correlation was found between gene expression and DNA methylation around transcription start sites (TSSs), while a positive correlation was observed in the gene body near TTSs. It was also possible that earlier demethylation of myogenic genes around TSSs contributes to their earlier expression in WZE. Using pyrosequencing to analyze DNA methylation patterns of promoter regions, we also found that earlier demethylation of the MyoD1 promoter in WZE contributed to its earlier expression. This study reveals that DNA demethylation of myogenic genes may contribute to embryonic leg muscle development differences between Wuzong and Shitou geese.

The regulation of granulosa cells (GCs) proliferation and apoptosis is the key step in follicular selection which determines the egg production performance of poultry. miR-202-5p has been reported to be involved in regulating the proliferation and apoptosis of mammalian ovarian GCs. However, its role in regulating the proliferation and apoptosis of goose GCs is still unknown. In the present study, the GCs of pre-hierarchical follicles (phGCs, 8-10 mm) and those of hierarchical follicles (hGCs, F2-F4) were used to investigate the role of miR-202-5p in cell proliferation and apoptosis during follicle selection. In phGCs and hGCs cultured in vitro, miR-202-5p was found to negatively regulate cell proliferation and positively regulate cell apoptosis. The results of RNA-seq showed that BTB Domain Containing 10 ( BTBD10 ) is predicted to be a key target gene for miR-202-5p to regulate the proliferation and apoptosis of GCs. Furthermore, it is confirmed that miR-202-5p can inhibit BTBD10 expression by targeting its 3'UTR region, and BTBD10 was revealed to promote the proliferation and inhibit the apoptosis of phGCs and hGCs. Additionally, co-transfection with BTBD10 effectively prevented miR-202-5p mimic-induced cell apoptosis and the inhibition of cell proliferation. Meanwhile, miR-202-5p also remarkably inhibited the expression of Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Beta ( PIK3CB) and AKT Serine/Threonine Kinase 1 ( AKT1), while it was significantly restored by BTBD10 . Overall, miR-202-5p suppresses the proliferation and promotes the apoptosis of GCs through the downregulation of PIK3CB / AKT1 signaling by targeting BTBD10 during follicular selection. Our study provides a theoretical reference for understanding the molecular mechanism of goose follicular selection, as well as a candidate gene for molecular marker-assisted breeding to improve the geese' egg production performance.

The meat and egg of goose is one of the main components of human food supply. The improvement of goose egg production is particularly important for the increasing human population. However, limited information is available about the effective molecular markers and mechanisms of egg production in goose. In this study, we jointly utilized the data of genome resequencing in different egg production Sichuan white goose and transcriptome at different follicle development stages to identified the molecular markers and mechanisms of egg production. The coefficient of variation of individual egg production in Sichuan white goose population is 0.42 to 0.49. Fifty individuals with the highest (laying 365 days egg number, LEN365 = 79-145) and 50 individuals with the lowest (LEN365 = 8-48) egg production were divided into high and low egg production groups. Based on whole-genome sequencing data of the selected samples, 36 SNPs (annotation novel.12.470, CELF2, ATP1A1, KCNJ6, RAB4A, UST, REV3L, DHX15, CAVN2, SLC5A9, Cldn5, MRPS23, and Tspan2) associated with the LEN365 were identified, involving multiple pathways such as metabolism and endocrinology. Notably, 5 SNPs located in the exon9 of ATP1A1 were identified by GWAS analysis. The association analysis with LEN365 showed the phenotypic variance explained of this haplotype consisting of 5 SNPs is 20.51%. Through transcriptome data analysis, we found the expression of ATP1A1 in the granular layers was increased in the stage of small yellow follicle to large yellow follicle (LYF) and LYF to F5, while decreased in F2 to F1. For the first time, we report the haplotype region formed by 5 SNPS on exon9 of ATP1A1 is associated with egg production in goose and involved in follicle selection and maturation processes., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

1. Due to seasonal breeding, geese breeds from Southern China have low egg yield. The genetic makeup underlying performance of local breeds is largely unknown, and few studies have investigated this problem. This study integrated 21 newly generated and 50 publicly existing RNA-seq libraries, representing the hypothalamus, pituitary and testis, to identify candidate genes and importantly related pathways associated with seasonal breeding in male Lion-Head geese.2. In total, 19, 119 and 302 differentially expressed genes (DEGs) were detected in the hypothalamus, pituitary and testis, respectively, of male Lion-Head geese between non-breeding and breeding periods. These genes were significantly involved in the neuropeptide signalling pathway, gland development, neuroactive ligand-receptor interaction, JAK-STAT signalling pathway, cAMP signalling pathway, PI3K-Akt signalling pathway and Foxo signalling pathway.3. By integrating another 50 RNA-seq samples 4, 18 and 40 promising DEGs were confirmed in hypothalamus, pituitary and testis, respectively.4. HOX genes were identified as having important roles in the development of testis between non-breeding and breeding periods of male Lion-Head geese.

Alternative products such as those from high-value protien animals have increased the demand for the production of high-quality chicken meat in past few years. This study examines the impact of two distinct feeding types on goose body-weight, as well as the genetic variation of growth hormone (GH) and pituitary-specific transcription factor (Pit-1) genes in ten goose populations using single nucleotide polymorphism (SNP) markers and PCR-RFLP analysis. Both genes were seen as very important for productivity, especially in light of the COVID-19 and its effect on poultry industry at the time. The findings suggest that employing genetic indicators in these two genes in conjunction with a high-fat diet may be a feasible strategy for goose selection programme aiming to increase marketing body weight, as the high-fat diet outperformed the balanced diet. The study investigates the effect of gender, 2 types of diets, breeds and the genetic variation of the two genes, four SNPs were reported to be found: two at the GH gene exons C123T and C158T, and two at the Pit-1 gene exons G161A and T282G. Certain genotypes were found to have a substantial effect on the marketing body-weight of goose, which varied depending on the tested breeds. However, in terms of gender, males report higher and better performance levels than females. Diet, breeds and genotype interaction, and breeds, gender and genotype interaction were found to have a minor effect on goose body weight. However, diet, breeds, gender, SNP locus, diet and breeds interaction, and breeds and gender interaction were found to have a significant effect on goose body weight, as indicated by the effect size results., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

1. The extracellular matrix (ECM) constitutes the basal lamina and the area between follicular cells. Remodelling the ECM is believed to be a key event in follicular development, especially during selection, and plays an important role in cell migration, survival, and steroidogenesis. miR-199-3p is differentially expressed in the goose granulosa layer during follicular selection and is reported to play a primary role in inhibiting cell migration and invasion. Nevertheless, the effect of miR-199-3p on ovarian follicles and its role in follicular cellular migration is not understood.2. In this study, qRT-PCR assays revealed that miR-199-3p was differentially expressed in the granulosa layer from goose ovarian follicles before and after follicular selection. Additionally, miR-199-3p overexpression in cultured granulosa cells (GCs) from goose pre-hierarchical follicles significantly suppressed cell viability and migration. It elevated the concentration of progesterone and the expression of key progesterone production genes. Furthermore, miR-199-3p overexpression in the GCs of goose pre-hierarchical follicles inhibited the expression of ECM-related genes ( ITGB8 , MMP9 and MMP15 ) yet promoted the expression of another two ECM-related genes ( COL4A1 and LAMA1 ). Finally, dual-fluorescence reporter experiments on 293T cells established the direct targeting of ECM gene ITGB8 by miR-199-3p.3. In conclusion, miR-199-3p may participate in granulosa cell migration, viability, and steroidogenesis in goose ovarian follicles before selection by modulating ITGB8 and other ECM-related genes.

Although miR-27b-3p has been evidenced to regulate the proliferation, apoptosis, and differentiation of a variety of mammalian cell types, its actions and mechanisms on ovarian cell steroidogenesis remains largely unknown in both mammalian and avian species. In this study, we aimed to determine the expression profiles of miR-27b-3p in granulosa cell layers during goose ovarian follicle development and to reveal its actions on estrogen (E 2 ) secretion of goose granulosa cells as well as the underlying regulatory mechanisms. It was observed that miR-27b-3p was ubiquitously expressed throughout follicle development but exhibited much higher levels in hierarchical- than in prehierarchical follicles. In cultured granulosa cells from the fourth through second largest preovulatory (F4-F2) follicles of goose, up- and downregulation of miR-27b-3p by using its mimic and inhibitor significantly decreased and increased E 2 secretion, respectively. Meanwhile, the mRNA levels of STAR and CYP19A1 were significantly reduced while those of CYP11A1 and 3βHSD were elevated in the mimic-transfected granulosa cells. By comparison, downregulation of miR-27b-3p enhanced the mRNA levels of STAR but had no significant effects on those of CYP19A1, CYP11A1, and 3βHSD. Results from bioinformatic prediction and luciferase reporter assay demonstrated that CYP1B1 was a downstream target of miR-27b-3p. Although the siRNA-mediated downregulation of CYP1B1 did not significantly change E 2 secretion by goose granulosa cells, it reduced the mRNA levels of STAR and CYP19A1 as well as those of LKB1 and AMPKα, which are involved in the AMPK signaling pathway. Taken together, these data suggest that miR-27b-3p plays an inhibitory role in E 2 secretion by goose F4-F2 granulosa cells, at least in part, by targeting CYP1B1 through the AMPK signaling pathway., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

The incubation behavior of geese seriously affects their egg production performance. Studies on incubation behavior have identified functional genes, but the regulatory architecture relationship between functional genes and chromatin accessibility remains poorly understood. Here, we present an integrated analysis of open chromatin profiles and transcriptome to identify the cis-regulatory element and their potential transcription factors involved in regulating incubation behavior in goose pituitary. Assay for transposase-accessible chromatin sequencing (ATAC-seq) revealed that open chromatin regions increased in the pituitary during the transition from incubation behavior to laying. We identified 920 significant differential accessible regions (DARs) in the pituitary. Compared to the laying stage, most DARs had higher chromatin accessibility in the brooding stage. Motif analysis of open DARs showed that the most significant transcription factor (TF) occupied sites predominantly enriched in motifs binding to the RFX family (RFX5, RFX2, and RFX1). While the majority of TF motifs enriched under sites of the nuclear receptor (NR) family (ARE, GRE, and PGR) in closed DARs at the incubation behavior stage. Footprint analysis indicated that the transcription factor RFX family exhibited higher binding on chromatin at the brooding stage. To further elucidate the effect of changes in chromatin accessibility on gene expression levels, a comparison of the transcriptome revealed 279 differentially expressed genes (DEGs). The transcriptome changes were associated with processes of steroid biosynthesis. By integrating ATAC-seq and RNA-seq, few DARs directly affect incubation behavior by regulating the transcription levels of genes. Five DAR-related DEGs were found to be closely related to maintaining the incubation behavior in geese. Footprinting analysis revealed a set of transcription factors (RFX1, RFX2, RFX3, RFX5, BHLHA15, SIX1, and DUX) which displayed the highest activity at the brooding stage. SREBF2 was predicted to be the unique differentially expressed transcription factor whose mRNA level was down-regulated and enriched in hyper-accessible regions of PRL in the broody stage. In the present study, we comprehensively profiled the transcriptome and chromatin accessibility in the pituitary related to incubation behavior. Our findings provided insight into the identification and analysis of regulatory elements in goose incubation behavior. The epigenetic alterations profiled here can help decipher the epigenetic mechanisms that contribute to the regulation of incubation behavior in birds.

Astroviruses are considered the cause of gastroenteritis in humans and animals. Studies in recent years show avian astroviruses are also associated with duckling hepatitis, gosling gout, and chicken nephritis. In this study, a GAstV strain, designated as JS2019/China, was detected in dead goslings from a commercial goose farm in Jiangsu province of China. Viral strain was proliferated in goose embryos and sequence analysis showed the isolated strain had a classical structure arrangement and a series of conserved regions compared with other GAstVs. Sequence comparison and phylogenetic analysis of whole genome and ORF2 revealed that JS2019/China belongs to the GAstV-1 group, which consists of most of the GAstV strains. Amino acid analysis indicated that some mutants might have an impact on viral protease capacity, such as V505I and K736E of ORF1a and T107I, F342S, and S606P of ORF2. Taken together, a novel GAstV strain was isolated and genomic analysis and protein polymorphism analysis indicated that some amino acid mutants might affect the viral virulence., (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

The development of follicles in the ovaries is a critical determinant of poultry egg production. There are existing studies on the follicular development patterns in poultry, but the specific regulatory mechanisms still need further study. In a previous study, we identified long non-coding RNA (lncRNA) MSTRG.5970.28, anosmin 1 (ANOS1), and its predicted target miR-133a-3p that may be associated with goose ovary development. However, the function of MSTRG.5970.28 in goose granulosa cells and its regulatory mechanisms affecting granulosa cell proliferation and apoptosis have not been reported. In the present study, MSTRG.5970.28 and miR-133a-3p overexpression and interference vectors were constructed. Combined with reverse-transcription real-time quantitative PCR (RT-qPCR), a dual luciferase activity assay, Cell Counting Kit-8 (CCK-8), and flow cytometric analysis, we investigated the role of the MSTRG.5970.28-miR-133a-3p-ANOS1 axis in goose follicular granulosa cells and the associated regulatory mechanisms. MSTRG.5970.28 was found to be localized in the cytoplasm and its expression was influenced by reproductive hormones. The targeting relationship among MSTRG.5970.28, ANOS1, and miR-133a-3p were verified by a dual luciferase activity assay. CCK-8 and apoptosis assays showed that MSTRG.5970.28 inhibited the proliferation and promoted apoptosis of goose granulosa cells. The regulatory role of miR-133a-3p on granulosa cell proliferation and apoptosis was opposite to MSTRG.5970.28. We found that the proliferative and apoptotic effects of granulosa cells caused by MSTRG.5970.28 overexpression were attenuated by miR-133a-3p. MSTRG.5970.28 functions as a competitive endogenous RNA that regulates ANOS1 expression by sponging miR-133a-3p and thus exerts regulatory functions in granulosa cells. In sum, the present study identified lncRNA MSTRG.5970.28 as associated with goose ovary development, which affects the expression of ANOS1 by targeting miR-133a-3p, thereby influencing the proliferation and apoptosis of goose granulosa cells., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The broodiness traits of domestic geese are a bottleneck that prevents the rapid development of the goose industry. To reduce the broodiness of the Zhedong goose and thus improve it, this study hybridized it with the Zi goose, which has almost no broody behavior. Genome resequencing was performed for the purebred Zhedong goose, as well as the F2 and F3 hybrids. The results showed that the F1 hybrids displayed significant heterosis in growth traits, and their body weight was significantly greater than those of the other groups. The F2 hybrids showed significant heterosis in egg-laying traits, and the number of eggs laid was significantly greater than those of the other groups. A total of 7,979,421 single-nucleotide polymorphisms (SNPs) were obtained, and three SNPs were screened. Molecular docking results showed that SNP11 located in the gene NUDT9 altered the structure and affinity of the binding pocket. The results suggested that SNP11 is an SNP related to goose broodiness. In the future, we will use the cage breeding method to sample the same half-sib families to accurately identify SNP markers of growth and reproductive traits., Competing Interests: The authors declare no conflict of interest.

The study aimed to determine the technological and sensory properties of the breast and thigh muscles of geese from the Polish native varieties: Kartuska (Ka) and Suwalska (Su) (from northern Poland) as well as Lubelska (Lu) and Kielecka (Ki) (from southern Poland). The color parameters: L*, a*, b*, ΔE, C, h°, total heme pigments (THPs), and share of myoglobin (Mb), metmyoglobin (MMb), and oxymyoglobin (MbO 2 ) in muscles were determined. In terms of technological properties, the following were determined: pH 24 , water-binding capacity (WBC), water-holding capacity (WHC), cooking (CL), and roasting losses (RL). In addition, a sensory evaluation of the raw meat color was performed. In roasted meat, a sensory evaluation and texture profile analysis (TPA) were carried out, as well as the shear force (SF) and chemical composition were determined. Roasted muscles of varieties native to northern Poland (Ka and Su) were higher in lipids (P≤0.05) than the muscles of southern varieties (Lu and Ki). Ka meat had the highest protein content, and Lu meat had the lowest (P≤0.05). The raw muscle color sensory evaluation results, the THP, and the L* and ΔE values indicated that the darkest color among the studied genotypes were the Ka muscles, and the lightest was Ki meat (P≤0.05). Lu's muscles are distinguished by better usability for processing and culinary purposes than the muscles of the other genotypes due to high pH 24 , WBC, WHC, and low RL and CL of thigh muscles, as well as high WHC and low RLs of the breast muscles (P≤0.05). Due to the tenderness, juiciness, and high general evaluation (P≤0.05), the best sensory features among the studied genotypes were found in the Ka breast and thigh muscles. The low SF value proved the higher tenderness of Ka geese muscles., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The thymus is a vital immune organ, but its function gradually declines with age. Circular RNAs (circRNAs) are related to the development of tissues and organs. In this study, bioinformatics analysis showed that 1329, 755, and 417 circRNAs were differentially expressed between the comparison groups of 6-month age (M6) and 20-embryo age (E20), 3-day post-hatch (P3), and 3-month age (M3) Magang geese, respectively. Among them, 167 circRNAs were differentially co-expressed between thymic development (E20, P3, and M3) and involution (M6). Functional analysis showed significant enrichment of phosphorylation and positive regulation of GTPase activity. Furthermore, pathway analysis has shown that glycerolipid metabolism and the Wnt signaling pathway are critical pathways in the thymic involution process. Finally, we constructed the competitive endogenous RNA (ceRNA) network. The results of this study suggest that circRNAs may be involved in the age-related thymic involution of the Magang goose., Competing Interests: Declaration of competing interest None., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Broodiness is an unfavorable trait associated with the cessation of egg laying. Studies have found that excessive granulosa cell apoptosis and autophagy occur during goose broodiness. Other studies have also confirmed that oxidative stress is an important cause of apoptosis and autophagy. However, whether oxidative stress occurs during goose broodiness and whether this oxidative stress causes apoptosis and autophagy have not been fully elucidated. In this study, we investigated the effects of oxidative stress on the autophagy and apoptosis of granulosa cells in broody geese. The results showed higher mRNA expression of genes related to antioxidative stress responses ( GPX , SOD-1 , SOD-2 , COX-2 , CAT and hsp70 ) in pre-broody and broody geese than in laying birds. In addition, increased levels of granulosa cell apoptosis and autophagy were observed in pre-broody geese than in laying geese. Additionally, granulosa cells treated with H 2 O 2 exhibited increased apoptosis and autophagy in vitro, and these effects were responsible for goose granulosa cell death. Moreover, vitamin E treatment effectively protected granulosa cells from H 2 O 2 -induced oxidative stress by inhibiting ROS production. Correspondingly, granulosa cell apoptosis and autophagy were greatly alleviated by vitamin E treatment. Together, our results demonstrated serious oxidative stress and granulosa cell apoptosis and autophagy in broody geese, and oxidative stress promoted apoptosis and autophagy. Vitamin E alleviated the autophagy and apoptosis of granulosa cells by inhibiting oxidative stress.

Background: Reconstructing phylogenetic relationships with genomic data remains a challenging endeavor. Numerous phylogenomic studies have reported incongruent gene trees when analyzing different genomic regions, complicating the search for a 'true' species tree. Some authors have argued that genomic regions of increased divergence (i.e. differentiation islands) reflect the species tree, although other studies have shown that these regions might produce misleading topologies due to species-specific selective sweeps or ancient introgression events. In this study, we tested the extent to which highly differentiated loci can resolve phylogenetic relationships in the Bean Goose complex, a group of goose taxa that includes the Taiga Bean Goose (Anser fabalis), the Tundra Bean Goose (Anser serrirostris) and the Pink-footed Goose (Anser brachyrhynchus)., Results: First, we show that a random selection of genomic loci-which mainly samples the undifferentiated regions of the genome-results in an unresolved species complex with a monophyletic A. brachyrhynchus embedded within a paraphyletic cluster of A. fabalis and A. serrirostris. Next, phylogenetic analyses of differentiation islands converged upon a topology of three monophyletic clades in which A. brachyrhynchus is sister to A. fabalis, and A. serrirostris is sister to the clade uniting these two species. Close inspection of the locus trees within the differentiated regions revealed that this topology was consistently supported over other phylogenetic arrangements. As it seems unlikely that selection or introgression events have impacted all differentiation islands in the same way, we are convinced that this topology reflects the 'true' species tree. Additional analyses, based on D-statistics, revealed extensive introgression between A. fabalis and A. serrirostris, which partly explains the failure to resolve the species complex with a random selection of genomic loci. Recent introgression between these taxa has probably erased the phylogenetic branching pattern across a large section of the genome, whereas differentiation islands were unaffected by the homogenizing gene flow and maintained the phylogenetic patterns that reflect the species tree., Conclusions: The evolution of the Bean Goose complex can be depicted as a simple bifurcating tree, but this would ignore the impact of introgressive hybridization. Hence, we advocate that the evolutionary relationships between these taxa are best represented as a phylogenetic network., (© 2023. The Author(s).)

Background: Geese exhibit relatively low reproductive performance, and follicular atresia is an important factor that restricts the egg production of geese. Systematic analysis of the regulation of follicle atresia in geese through transcriptome and proteome levels could provide meaningful information on clarifying the mechanism of follicle atresia in poultry., Result: The granulosa cell layer was loose, disintegrated and showed apoptosis in atretic follicles and remained intact in normal follicles. The hormone levels of FSH and LH were significantly decreased in the atresia follicles compared to the normal follicles (P < 0.05). A total of 954 differentially expressed genes (DEGs, 315 increased and 639 decreased) and 161 differentially expressed proteins (DEPs, 61 increased and 100 decreased) were obtained in atresia follicles compared to normal follicles, of which, 15 genes were differentially expressed in both transcriptome and proteome. The DEGs were mainly enriched in sodium transmembrane transport, plasma membrane, and transmembrane transporter activity based on the GO enrichment analysis and in the cell cycle pathway based on the KEGG enrichment analysis. The DEPs were mainly enriched in localization, lysosome, and phospholipid-binding based on the GO enrichment analysis. Candidate genes Smad2/3, Smad4, Annexin A1 (ANXA1), Stromelysin-1 (MMP3), Serine/threonine-protein kinase (CHK1), DNA replication licensing factor (MCM3), Cyclin-A2 (CCNA2), mitotic spindle assembly checkpoint protein (MAD2), Cyclin-dependent kinase 1 (CDK1), fibroblast growth factor 12 (FGF12), and G1/S-specific cyclin-D1 (CCND1) were possibly responsible for the regulation of atresia., Conclusion: The cell cycle is an important pathway for the regulation of follicular atresia. Sodium outflow and high expression of MMP3 and MMP9 could be responsible for structural destruction and apoptosis of follicular cells., (© 2023. The Author(s).)

The development of mammalian nonalcoholic fatty liver disease is associated with oxidative stress, reduced mitochondrial function, and increased apoptosis in hepatocytes; however, the expressions of mitochondria-related genes are elevated in goose fatty liver, suggesting that there may be a unique protective mechanism in goose fatty liver. The aim of the study was to investigate this protective mechanism in terms of anti-oxidant capacity. Our data showed no substantial differences in the mRNA expression levels of the apoptosis-related genes including B-cell lymphoma-2 (Bcl-2), BCL2-associated X (Bax), cysteinyl aspartate-specific proteinase-3 (Caspase-3), and cysteinyl aspartate-specific proteinase-9 (Caspase-9) in the livers of the control and overfeeding Lander geese groups. The protein expression levels of Caspase-3 and cleaved Caspase-9 were not markedly different between the groups. Compared with the control group, malondialdehyde content was significantly lower (P < 0.01), glutathione peroxidase (GSH-Px) activity, glutathione (GSH) content, and mitochondrial membrane potential levels were higher (P < 0.01) in the overfeeding group. The mRNA expression levels of the anti-oxidant genes superoxide dismutase 1 (SOD1), glutathione peroxidase 1 (GPX1), and glutathione peroxidase 2 (GPX2) were increased in goose primary hepatocytes after 40 mM and 60 mM glucose treatment. Reactive oxygen species (ROS) levels were significantly reduced (P < 0.01), whereas the mitochondrial membrane potential was maintained at normal levels. The mRNA expression levels of the apoptosis-related genes Bcl-2, Bax, and Caspase-3 were not substantial. There were no significant differences in the expression levels of Caspase-3 and cleaved Caspase-9 proteins. In conclusion, glucose-induced enhanced anti-oxidant capacity may help protect the function of mitochondria and inhibit the occurrence of apoptosis in goose fatty liver., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Poultry is subject to varying degrees of feather loss and feather pecking during production, which seriously affects the live appearance and carcass appearance of their commercial traits and greatly reduces the production profitability of the farming enterprise. It also has an impact on down production and quality in the case of geese. In this study, mathematical models (Logistic, Gompertz, and Von Bertalanffy) were used to assess feather growth and development during the embryonic period in Jilin white geese (Anser cygnoides) predicting the weight and length of feathers from the back, chest, and belly tracts at different embryonic ages, to determine which growth model more accurately described feather growth patterns. The result first showed that the primary feather follicles of the Jilin white goose developed at E14 and secondary feather follicles at E18; primary feather follicle density increased and then decreased, whereas secondary feather follicle density increased continuously and the primary and secondary feather follicles developed independently. Secondly, the embryonic feather growth followed a slow-fast-slow pattern, with feathers growing slowly from E12 to E18, quickly from E18 to E24, and then decreasing after E24 until just before emergence (E30). In addition, before E14, feathers were concentrated in the back tracts, and no feathers were found on the head, neck, chest, abdomen, or wings. By E22, the whole body of the embryo was covered with feathers, and the back feathers were the earliest and fastest to develop. Compared to the Gompertz and von Bertalanffy models, the logistic model fit (R2 = 0.997) was the highest, while the sum of residual squares (RSS = 25661.67), Akaike's information criterion (AIC = 77.600), Bayesian information criterion (BIC = 78.191), and mean square error (MSE = 2851.296) were the lowest. Therefore, the logistic model was more suitable for describing the changes in whole-body feather growth during the embryonic period in Jilin white geese. In conclusion, using the growth curve model to explain the relationship between feather growth and embryonic age in geese will potentially speed up the process of genetic improvement in Jilin white geese (A. cygnoides) and thus provide scientific support for molecular genetic breeding., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Granular cells proliferation in goose regulated by bone morphogenetic proteins (BMPs) signaling pathway is still unknown. In this experiment, BMPs and their receptor, and receptor activated mothers against decapentaplegic homologs (SMADs) were quantitatively expressed in granular cell layer of pre-hierarchycal and hierarchycal follicles in Wanxi White goose. The screened BMP was then used for construction of overexpressed and knockdown vectors and transfected into granular cells of goose to assess the cell proliferation and apoptosis. Granular cells with BMP-overexpressed were then used for ChIP-Seq analysis to elucidate the molecular mechanism of BMP affecting granular cell proliferation. The results showed that the mRNA expression of BMP4 was significantly expressed in pre-hierarchical follicles, and also highly expressed in hierarchical follicles than other BMPs, while the Ⅰ and Ⅱ type of BMP receptors were expressed in basic level. The mRNA expression of SMAD8 was significantly elevated in pre-hierarchical follicles. Overexpression of BMP4 could promote the proliferation of granular cells and inhibited the expression of BMP4 caused a higher cell apoptosis. ChIP-Seq identified multiple regulatory targets of SMAD4, which were mostly related to cell cycle and lipid metabolism according to the GO and KEGG pathway enrichment. From the five most significant binding motif and quantitative expression verification, the activin membrane binding inhibitor (BAMBI) was down regulated in BMP4 overexpressed granular cells. In conclusion, the BMP4 was highly expressed in granular cells and phosphorylates SMAD8, the activated SMAD8 combined with SMAD4 transfers into nucleus to regulate the expression of BAMBI to promote lipid synthesis., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Lingxian white goose (LXW) is a goose breed indigenous to China that is famous for its meat quality and fast growth. However, the genomic evidence underlying such excellent breeding characteristics remains poorly understood. Therefore, we performed whole-genome resequencing of 141 geese from 3 indigenous breeds to scan for selection signatures and detect genomic regions related to breed features of LXW. We identified 5 reproduction-related genes (SYNE1, ESR1, NRIP1, CCDC170, and ARMT1) in highly differentiated regions and 11 notable genes in 26 overlapping windows, some of which are responsible for meat quality (DHX15), growth traits (LDB2, SLIT2, and RBPJ), reproduction (KCNIP4), and unique immunity traits (DHX15 and SLIT2). These findings provide insights into the genetic characteristics of LXW and identify genes affecting important traits in LXW, which extends the genetic resources and basis for facilitating genetic improvement in domestic geese breeds., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Anatidae contains numerous waterfowl species with great economic value, but the genetic diversity basis remains insufficiently investigated. Here, we report a chromosome-level genome assembly of Lion-head goose (Anser cygnoides), a native breed in South China, through the combination of PacBio, Bionano, and Hi-C technologies., Findings: The assembly had a total genome size of 1.19 Gb, consisting of 1,859 contigs with an N50 length of 20.59 Mb, generating 40 pseudochromosomes, representing 97.27% of the assembled genome, and identifying 21,208 protein-coding genes. Comparative genomic analysis revealed that geese and ducks diverged approximately 28.42 million years ago, and geese have undergone massive gene family expansion and contraction. To identify genetic markers associated with body weight in different geese breeds, including Wuzong goose, Huangzong goose, Magang goose, and Lion-head goose, a genome-wide association study was performed, yielding an average of 1,520.6 Mb of raw data that detected 44,858 single-mucleotide polymorphisms (SNPs). Genome-wide association study showed that 6 SNPs were significantly associated with body weight and 25 were potentially associated. The significantly associated SNPs were annotated as LDLRAD4, GPR180, and OR, enriching in growth factor receptor regulation pathways., Conclusions: We present the first chromosome-level assembly of the Lion-head goose genome, which will expand the genomic resources of the Anatidae family, providing a basis for adaptation and evolution. Candidate genes significantly associated with different goose breeds may serve to understand the underlying mechanisms of weight differences., (© The Author(s) 2023. Published by Oxford University Press GigaScience.)

Background: Hungarian white goose has excellent down production performance and was introduced to China in 2010. The growth and development of feather follicles has an important impact on down production. Goose feather follicles can be divided into primary and secondary feather follicles, both of which originate in the embryonic stage. Msx2 (Msh Homeobox 2) plays a regulatory role in tissues and organs such as eyes, teeth, bones and skin. However, its regulatory mechanism on goose feather follicles development remains unclear., Results: Msx2 gene first increased, then decreased and increased at the end (E13, E18, E23, E28) during embryonic feather follicle development, and the expression level was the highest at E18. The pEGFP-N1-Msx2 overexpression vector and si-Msx2 siRNA vector were constructed to transfect goose embryo dermal fibroblasts. The results showed that the cell viability of ov-Msx2 group was significantly increased, and the gene expression levels of FGF5 and TGF-β1 genes were significantly down-regulated (P < 0.05), the expressions of PCNA, Bcl2, CDK1, FOXN1 and KGF genes were significantly up-regulated (P < 0.05). After transfection of siRNA vector, the cell viability of the si-Msx2 group was significantly decreased (P < 0.01) compared with the si-NC group. TGF-β1 expression was significantly up-regulated (P < 0.05), FGF5 expression was extremely significantly up-regulated (P < 0.01), while PCNA, Bcl2, CDK1, FOXN1 and KGF gene expression was significantly down-regulated (P < 0.05). High-throughput sequencing technology was used to mine the exon SNPs of Msx2. A total of 11 SNP loci were screened, four of the SNPs located in exon 1 were missense mutations. The feather follicle diameter of the GC genotype at the G78C site is significantly larger than that of the other two genotypes., Conclusions: Msx2 maybe inhibit the apoptosis of goose dermal fibroblasts and promotes their proliferation. G78C can be used as a potential molecular marker for downy Variety., (© 2022. The Author(s).)

Light is a factor affecting muscle development and meat quality in poultry production. However, few studies have reported on the role of light in muscle development and meat quality in geese. In this experiment, 10 healthy 220-day-old Zhedong white geese were reared for 60 d under a long photoperiod (15L:9D, LL) and short photoperiod (9L:15D, SL). The gastrocnemius muscles were collected after slaughter to evaluate muscle fiber characteristics and meat color, and RNA-seq analysis. The results showed that compared to the LL group, the SL group had large muscle fiber diameter and cross-sectional area, few muscle fibers per unit area, high meat color a* value, and low L* value at 24 h postmortem. On comparing the 2 groups, 70 differentially expressed genes (DEGs) were identified. Compared to the SL group, the LL group had 25 upregulated and 45 downregulated genes. Gene Ontology (GO) enrichment analysis showed that these DEGs were mainly involved in cell, cell part, binding, cellular processes, and single-organism processes. Several significantly enriched athways were identified in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, such as the calcium and PI3k-Akt signaling pathways. The expression of five randomly selected DEGs was verified using quantitative real-time PCR, and the results were consistent with the RNA-seq data. This study provides a theoretical basis for studying the molecular mechanisms by which light affects muscle development and meat color in geese., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The swan goose (Anser cygnoides) is the ancestor of the Chinese domestic goose. A previous study reported a scaffold-level genome version for a Chinese indigenous goose breed, and this assembly was used as the swan goose's reference genome. To date, there is still a lack of a chromosome-level genome for the swan goose. Here, we reported a de novo assembly of the genome of a wild swan goose using an integrated strategy that combines Illumina Hiseq, Oxford Nanopore and chromosome conformation capture (Hi-C) sequencing. A total of 134.6 Gb Nanopore data with sequencing coverage of 110.33 and 69.45 Gb Illumina data with coverage of 56.93 were obtained. The genome assembly size was 1153.41 Mb, with a contig N50 of 22.75 Mb. The total size and N50 length of our assembly were larger than the previously reported scaffold-level genome version. In addition, whole-genome sequencing data of 10 geese were mapped to the previous and the current assemblies. On average, 97.88 and 93.18% of the reads were properly mapped and paired into our and the previous assemblies. This high-quality chromosome-level swan goose genome could provide a valuable resource for the utilisation of goose studies and breeding., (© 2022 Stichting International Foundation for Animal Genetics.)

1. The gene IGF2BP1 has been reported as being related to the body size and body weight in ducks, goats and chickens. However, this potential growth-related gene has not yet been reported in geese.2. The goose IGF2BP1 cDNA (IGF2BP1-X1) is 2,925 bp in length, containing an open reading frame (ORF) of 1,731 bp that encodes a protein of 576 amino acids. Six IGF2BP1 transcripts (IGF2BP1-X2∼IGF2BP1-X7) were identified due to the alternative splicing of different exons within the IGF2BP1-X1 transcript.3. RT-qPCR analysis indicated that the goose IGF2BP1 mRNA was differentially expressed in the examined tissues of female embryos (28 d of development), adult (70 d of age) and laying (270 d of age) Zhedong White geese.4. In total, 16 single nucleotide polymorphisms (SNPs) and three insertion/deletion (InDel) variants were detected in several introns and 3'-untranslated regions of the goose IGF2BP1 gene. The 17-bp InDel within IGF2BP1 intron 14 was significantly associated with body weight at six weeks old (BW6, P < 0.05), and extremely significantly associated with the BW8 and BW10 (P < 0.01) of Zhedong White geese.

1. A total of 772, 420-day-old Xingguo grey geese (XGG) were sequenced using a low-depth (~1 x) whole-genome resequencing strategy to reveal the genetic mechanism of wing length-related traits by genome-wide association analysis (GWAS).2. The results showed that 119 SNPs had genome-wide significance for wing length in five regions of chromosome 4, of which the most significant locus (P = 7.95E-11) was located upstream of RBM47 and explained 7.3% of the phenotypic variation.3. A total of 219 SNPs located on chromosome 4 were associated with 2-joint-wing length, of which four SNPs reached the genome-wide significant level. However, for the length of 1-joint-wing and primary feather, we did not detect any associated locus.4. Six promising candidate genes, RBM47, SLAIN2, GRXCR1, SLC10A4, APBB2 and NSUN7 on chromosome 4, may play an important role in the growth and development of feathers, muscles and bones.

1. This study measured six reproduction traits in a Sichuan white goose population (209 individuals), including fertility, qualified egg rate, plasma concentrations of progesterone (P), follicle-stimulating hormone (FSH), prolactin (PRL) and oestrogen (E2).2. Whole-genome resequencing data from the same goose population (209 individuals) were used in a genome-wide association study (GWAS) utilising a mixed linear model to investigate the genes and genetic markers associated with reproduction traits. The frequency of the selected SNPs and haplotypes were determined using the Matrix-Assisted Laser Desorption Ionisation Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) method.3. In total, 42 SNPs significantly associated with these traits were identified. A haplotype block was constructed based on five SNPs that were significantly associated with qualified egg rate, with individuals having the haplotype CCTTAAGGAA having the lowest qualified egg rate.4. In conclusion, these results provided potential markers for marker-assisted selection to improve goose reproductive performance and a basis for elucidating the genetics of goose reproduction.

Fatal gout in geese caused by goose astrovirus (GAstV) has been spreading rapidly in China since 2018, causing serious economic losses in the goose breeding industry. To achieve simple, convenient and sensitive detection of GAstV, a novel diagnostic test was developed by combining reverse transcription-enzymatic recombinase amplification (RT-ERA) and CRISPR-Cas12a technologies. RT-ERA primers were designed to pre-amplify the conserved region of the ORF2 gene of GAstV and the predefined target sequence detected using the Cas12a/crRNA complex at 37℃ for 30 min. Specific detection of GAstV was achieved with no cross-reaction with non-GAstV templates and a sensitivity detection limit of 2 copies. The experimental procedure could be completed within 1 h, including RNA extraction (15 min), RT-ERA reaction (20 min), CRISPR-Cas12a/crRNA detection (5 min) and result readout (within 2 min) steps. In conclusion, the combination of RT-ETA and CRISPR-Cas12a provides a rapid and specific method that should be effective for the control and surveillance of GAstV infections in farms from remote locations., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Feather follicles and scales are two types of skin appendages distributed on different parts of avian skin. The morphogenesis and development of scales in waterfowl remain largely unknown. Here, we used H&E staining, ISH and RNA sequencing to reveal the morphological and molecular variations at the early development of scutate scales in goose shank skin. Transcriptome analysis produced 1824 differentially expressed genes (DEGs) regulating the induction of scales and further enriched gene function in cell adhesion and Wnt signaling pathway, etc. A total of 8 candidate genes (ALDOC, CSRP2, KRT15, KRT75, LGALS1, S100A6, OGN and SFRP2) were further detected by RT-qPCR to show upregulated (6 genes) and downregulated (2 genes) from pre-placodal to placode stage during the induction of goose scales. The localization of 7 candidate genes (ALDOC, CSRP2, CD109, KRT15, KRT75, S100A6, and OGN) by ISH suggests the potential roles for dermal and epidermal development during the induction of scutate scales. The dynamic molecular changes and specific gene expression patterns revealed in this report provide general knowledge of scale development in waterfowl as well as skin appendage diversity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

FASN plays a critical role in lipid metabolism, which is involved in regulating ovarian follicular development. However, the molecular mechanisms of how FASN regulate the function of ovarian follicular cells still remain elusive. In this study, by overexpression or interference of FASN in pre-hierarchical follicle granulosa cells (phGCs) and hierarchical follicle granulosa cells (hGCs), we analyzed their effects on the granulosa cell transcriptome and metabolome profiles using RNA-Seq and LC-MS/MS, respectively. The results showed that overexpression of FASN promoted proinflammatory factors expression by activating TLR3 / IRF7 and TLR3 / NF-κB pathways in phGCs, but only by activating TLR3 / IRF7 pathways in hGCs. Then, necroptosis and apoptosis were triggered through the JAK / STAT1 pathway (induced by inflammatory factors) and BAK / caspase-7 pathway, respectively. The combined analysis of the metabolome and transcriptome revealed that FASN affected the demand of GCs for 5-hydroxytryptamine (5-HT) by activating the neuroactive ligand-receptor interaction pathway in two categorized GCs and only altering the metabolic pathway of tryptophan in phGCs, and ultimately participated in regulating the physiological function of geese GCs. Taken together, this study showed that the mechanisms of FASN regulating the physiological function of geese phGCs and hGCs were similar, but they also had some different characteristics.

Geese are herbivorous birds that play an essential role in the agricultural economy. We construct the chromosome-level genome of a Chinese indigenous goose (the Xingguo gray goose, XGG; Anser cygnoides) and analyze the adaptation of fat storage capacity in the goose liver during the evolution of Anatidae. Genomic resequencing of 994 geese is used to investigate the genetic relationships of geese, which supports the dual origin of geese (Anser cygnoides and Anser anser). Chinese indigenous geese show higher genetic diversity than European geese, and a scientific conservation program can be established to preserve genetic variation for each breed. We also find that a 14-bp insertion in endothelin receptor B subtype 2 (EDNRB2) that determines the white plumage of Chinese domestic geese is a natural mutation, and the linkaged alleles rapidly increase in frequency as a result of genetic hitchhiking, leading to the formation of completely different haplotypes of white geese under strong artificial selection. These genomic resources and our findings will facilitate marker-assisted breeding of geese and provide a foundation for further research on geese genetics and evolution., (© 2022. The Author(s).)

Bar-headed geese (Anser indicus) are adaptable to plateau environments. In this study, we sequenced and assembled a high-quality chromosome-level genome of the bar-headed goose using PacBio long reads and Hi-C technique, and generated 115.73 Gb of Illumina short-reads and 95.89 Gb of PacBio long-reads. The assembled bar-headed goose genome, with a contig N50 of 5.734 Mb and a scaffold N50 of 65.77 Mb, is 1.129 Gb in length and includes 33 chromosomes and 451 fragments. BUSCO assessment yielded a completeness score of 94.4%. In total, 15,376 protein-coding genes were predicted, of which 94.95% had homologs in protein databases. We identified 78 positively selected genes (PSGs) in the bar-headed goose genome, which were mainly enriched in calcium ion and ATP-binding. This bar-headed goose genome will be an important resource for increasing our understanding regarding the genetic basis of adaptation to life at a high altitude., (© 2022. The Author(s).)

Goose ( Anser cygnoides ) are highly broody with low egg production, and large number of granulosa cells undergo apoptosis during broodiness. Our previous study has found that miR-34c-5p was highly abundant in the ovary of goose with broodiness phenotype. However, the mechanism that miR-34c-5p regulates granulosa cells function remains unclear. Here, we demonstrated that broody goose had higher levels of miR-34c-5p than that in laying goose by qRT-PCR. The dual luciferase reporter assay showed that Bcl2 was identified as a direct target of miR-34c-5p , which could be negatively regulated by miR-34c-5p . Furthermore, over-expression of miR-34c-5p significantly increased the rate of apoptosis and slowed down the proliferation of granulosa cells by inhibiting the Bcl2 expression, whereas the opposite trend was obtained when granulosa cells were supplemented with miR-34c-5p inhibitors. In addition, Bcl2 mRNA level was lower in goose with a brooding phenotype than that in goose with an egg-laying phenotype. Taken together, the data suggested that miR-34c-5p regulated granulosa cells apoptosis and brooding behavior by targeting Bcl2 , which not only contribute to reveal the potential mechanism of miR-34c-5p underlying granulosa cells apoptosis in goose, but also provides an effective strategy to reduce the incidence of broodiness and improve the egg production.

The laying performance of geese is mainly determined by follicular development and atresia, while follicular status is regulated by photoperiod. To understand the effect of photoperiod on the development of goose follicles, artificial light was used to change the photoperiod. In this study, ten healthy 220-day-old Zhedong white geese (Anser Cygnoides) with similar body weights and similar reproductive start times were reared for 60 days under long photoperiod (15 L:9 D) and short photoperiod (9 L:15 D) artificial light with the intensity controlled at 30 lux, and follicles were collected. Follicle development was analyzed by observing the morphology of follicle tissue, the localization of autophagosomes and autolysosomes, and the expression levels of apoptosis-related protein factors. Small white follicles (SWFs) were selected for RNA sequencing and bioinformatics analysis of the transcriptome. Under a long photoperiod, microtubule-associated protein 1 light chain 3 (LC3) and Caspase-3 were expressed in the granulosa cell layer and oocytes, respectively. LC3 and Caspase-3 protein expression was increased in SWF and large white follicles (LWFs), and there were more autophagosomes and autolysosomes in granulosa cells. RNA-seq found 93 differentially expressed genes (DEGs) in the short-photoperiod group, including 55 upregulated DEGs and 38 downregulated DEGs, distributed in 37 gene ontology categories. Kyoto Encyclopedia of Genes and Genomes-enriched signaling pathways revealed 5 pathways enriched in upregulated DEGs, including protein digestion and absorption, ECM-receptor interaction and regulation of lipolysis in adipocytes, and 4 pathways enriched in downregulated DEGs, such as fatty acid biosynthesis. Ten differentially expressed genes related to extracellular matrix and fatty acid metabolism (THBS2, COL12A1, MRC2, TUBA, COL1A1, COL11A1, HSPG2, FABP, MGLL, and OLAH) may be involved in the photoperiod regulation of follicle development in Zhedong white geese. The differentially expressed genes screened in this study will provide new ideas to further understand the molecular mechanism underlying photoperiod-mediated regulation of follicle development in Zhedong white geese., (Copyright © 2022. Published by Elsevier Inc.)

Background: Ovarian development is regulated by genes that are expressed dynamically and stage-specifically. Circular RNA (circRNA) has been proven to play a significant role in the regulation of animal reproduction., Objective: Studying the expression characteristics of circRNAs in goose ovaries at various egg-laying stages can provide a reference for the molecular regulation mechanism of ovary development in geese that is mediated by circRNAs., Methods: In this study, the expression profiles of circRNAs were compared in ovary tissues from Yili geese in three different breeding periods, namely the prelaying period (KL), laying period (CL), and ceased period (XL), and differentially expressed circRNAs related to ovarian development in Yili geese were screened. The potential biological functions of differential circRNAs were predicted by bioinformatics, and the differential circRNA-miRNA regulatory network was constructed., Results: The results showed that a total of 4483 circRNAs were identified in 12 ovarian tissue samples from Yili geese at different laying stages. In the KL vs. CL, XL vs. CL, and XL vs. KL groups, 159, 455, and 383 differentially expressed circRNAs were identified, respectively. The host genes of the differential circRNAs were mostly enriched in the signal transduction, metabolism, and other related pathways, such as those for phototransduction, glycerophospholipid metabolism, aminoacyl-tRNA biosynthesis, and retinol metabolism. Finally, we constructed circRNA-miRNA regulation networks. Nine differential circRNAs were randomly selected for qRT-PCR verification, and the expression trends were consistent with the sequencing results., Conclusions: Our results indicated that significant differences in the expression profiles of circRNAs in the ovaries of Yili geese at different egg-laying stages. Meanwhile, through analyzing the differential circRNA-miRNA interaction network, core regulators such as circRNA NW&#95;013186107.1:36835|52,574 and gga-miR-34b-5p were screened. This study provides a reference for the further analysis of the molecular regulatory mechanism of the circRNAs regulating goose ovary development and enriches the theory of genetic regulation during goose ovary development., (© 2022. The Author(s) under exclusive licence to The Genetics Society of Korea.)

In recent years, an infection in geese caused by goose astrovirus (GAstV) has repeatedly occurred in coastal areas of China and rapidly spread to inland provinces. The infection is characterized by joint and visceral gout and is fatal. The disease has caused huge economic losses to China's goose industry. GAstV is a nonenveloped, single-stranded, positive-sense RNA virus. As it is a novel virus, there is no specific classification. Here, we review the current understanding of GAstV. The virus structure, isolation, diagnosis and detection, innate immune regulation, and transmission route are discussed. In addition, since GAstV can cause gout in goslings, the possible role of GAstV in gout formation and uric acid metabolism is discussed. We hope that this review will inform researchers to rapidly develop effective methods to prevent and treat this disease., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this article., (Copyright © 2022 Chenggang Liu et al.)

Background: The development of the ovaries is an important factor that affects egg production performance in geese. Ovarian development is regulated by genes that are expressed dynamically and stage-specifically. The transcriptome profile analysis on ovarian tissues of goose at different egg laying stages could provide an important basis for screening and identifying key genes regulating ovarian development., Results: In this study, 4 ovary tissues at each breeding period of pre-laying (PP), laying (LP), and ceased-laying period (CP), respectively, with significant morphology difference, were used for RNA extraction and mRNAs, lncRNAs, and miRNAs comparison in Yili geese. CeRNA regulatory network was constructed for key genes screening. A total of 337, 1136, and 525 differentially expressed DE mRNAs, 466, 925, and 742 DE lncRNAs and 258, 1131 and 909 DE miRNAs were identified between PP and LP, between CP and LP, and between CP and PP groups, respectively. Functional enrichment analysis showed that the differentially expressed mRNAs and non-coding RNA target genes were mainly involved in the cell process, cytokine-cytokine receptor interaction, phagosome, calcium signaling pathway, steroid biosynthesis and ECM-receptor interaction. Differential genes and non-coding RNAs, PDGFRB, ERBB4, LHCGR, MSTRG.129094.34, MSTRG.3524.1 and gga-miR-145-5p, related to reproduction and ovarian development were highly enriched. Furthermore, lncRNA-miRNA-mRNA regulatory networks related to ovary development were constructed., Conclusions: Our study found dramatic transcriptomic differences in ovaries of Yili geese at different egg-laying stages, and a differential lncRNA-miRNA-mRNA regulatory network related to cell proliferation, differentiation and apoptosis and involved in stromal follicle development were established and preliminarily validated, which could be regarded as a key regulatory pathway of ovarian development in Yili geese., (© 2022. The Author(s).)

Although graylag geese (A. anser) showed similar plumages of white, grey, and white with grey patches compared to those in swan geese (A. cygnoides), it was believed the substantial molecular mechanism for plumage variations were different. To date, studies on genes responsible for diverse plumages among graylag geese were limited and causal mutations remain unknown. In this study, genomes from 57 individuals belonging to six breeds showing different plumages were sequenced at ∼10X depth. Firstly, the allele frequency differences (AFD) of variants on the scaffold394 (NW&#95;013185915.1) between grey and white goose breeds (A. anser) was calculated and a genomic region between 768,290-779,889 bp was detected to carry candidate variants associated with plumages, including one SNP (g. 775,151G > T, ∼18.6 kb upstream of EDNRB2) found to be fixed in white geese. This region was overlapped with the one detected by the haplotype-based sweep analysis, in which significant signals defined a candidate region of 736,610-820,622 bp on the same scaffold. Results from the transcriptomic data showed that expression levels of EDNRB2 and many other melanogenesis-related genes were significantly decreased among white geese compared to that in grey geese, especially at late embryonic stages (>E15). Modifications at transcriptional levels might result in abnormal melanocyte developments and thus the white plumages when they grow up. In addition, a frameshift mutation (C > -) in exon4 of MLANA gene on scaffold176 (NW&#95;013185876.1) was suggested as the causal mutation for sex-linked dilution phenotype in graylag geese although this requires more demonstration experiments. Together with observed white plumages caused by EDNRB2 mutations in coding regions among swan geese and chicken, our study provided new examples to study the parallel evolution., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

Lysozymes, acting as antimicrobial molecules, play a vital role in the host's innate immune response to pathogen infections. In the present study, a g-type lysozyme gene termed Tf-LyzG from roughskin sculpin, Trachidermus fasciatus was firstly reported. The deduced amino acid sequence of Tf-LyzG contained 188 residues and possessed conserved catalytic residues (Glu 71 , Asp 84 , and Asp 95 ). Gene expression analysis revealed that Tf-LyzG was widely distributed in the tested eleven tissues with the highest expression in the gill and could be significantly induced post lipopolysaccharide (LPS) challenge. The lysozyme activity of the purified recombinant protein (rTf-LyzG) was found to be most active at pH 5.5 and 37 °C. rTf-LyzG exhibited a wide spectrum of potent bacteriolytic activity against four Gram-positive bacteria and six Gram-negative bacteria. It also displayed a high affinity to polysaccharides on bacteria surfaces including LPS, lipoteichoic acid (LTA), and peptidoglycan (PGN). rTf-LyzG was capable of binding and agglutinating all nine bacteria. Flow cytometry assay further revealed that rTf-LyzG could disrupt the membrane of Micrococcus lysodeikticus which is confirmed by scanning electron microscope (SEM) analysis that reveals blebs around the bacterial cell membrane. In summary, these data indicate that Tf-LyzG is of great importance in the fish immune response against pathogens invasion., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

1. The objective of this study was to compare carcase weight and composition and meat quality traits of Landes, Slovak White and Kuban geese after four reproductive seasons. A total of 42 goose carcases (seven males and seven females from each breed) were assessed.2. After four reproductive seasons, the breeds differed in eviscerated carcase weight. Slovak White geese showed more breast muscle with a cooking loss, whereas Kuban geese had a higher percentage of neck and carcase discards compared to the other breeds. The breeds differed in the chemical composition of breast and leg muscles, except for collagen in breast muscles and protein in leg muscles.3. Regardless of breed, males had heavier carcase weight and higher breast muscle, leg muscle, neck and wings and lower percentages of skin with subcutaneous fat and abdominal fat relative to body weight compared to females. Sex had a significant effect on the fat and salt content, meat lightness, most textural characteristics and endomysium thickness of breast muscle. The same differences in leg muscles for chemical composition (except for protein percentage) and lightness were recorded.4. Breed x sex interactions were significant for the proportion of leg muscle, chemical composition of breast and leg muscles (except for leg protein content), muscle fibre cross-sectional area and fibre perimeter of pectoralis major muscle.5. The results showed that the carcases and meat of the goose breeds after four reproductive seasons were suitable for consumers and meat processors, due to favourable composition, high nutritive value and good technological properties.

Since 2017, outbreaks of gosling astroviruses (GoAstV) causing the major symptoms related to gout in geese have posed a threat to China's poultry industry and caused huge economic losses. In this study, tissue samples from goslings with gout and urate deposition as the main symptoms were taken from 14 goose farms in different regions of China and screened for pathogen infection. The infection rate of GoAstV was 100%, whereas the infection rates of goose parvovirus, reovirus, Tembusu virus, and goose hemorrhagic polyomavirus were 2, 4, 0, and 0%, respectively. In total, 14 GoAstV strains were isolated and their complete genomes were sequenced. Based on the phylogenetic trees, the 14 isolated strains were classified as GoAstV (G-I) and were considered distant from strains belonging to GoAstV (G-II). The multiple sequence alignments indicated a tremendous amount of amino acid mutations in some parts of the encoding proteins of these strains; the main mutations were located in open reading frames (ORFs)-ORF1a and ORF2, such as M533V and F568S in ORF1a and A614T in ORF2. On the other hand, Further, 2 of the 14 GoAstV strains were possibly derived through inter-GoAstV-I recombination. Taken together, these findings indicate that GoAstVs are evolving in a more complex manner and have diverse transmission routes., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Egg-ceasing is a phenomenon that occurs in most avian species and significantly reduces productivity. Although several factors are reported to regulate the reproduction progress, the underlying molecular mechanism of egg-ceasing remains obscure. Herein, we identified and explored the differentially expressed miRNAs and mRNAs involved in ovarian atrophy via high throughput sequencing. We identified a total of 901 mRNAs and 50 miRNAs that were differentially expressed in egg-laying and atrophic ovaries. Among them, numerous differentially expressed gene (DEG) transcripts and target genes for miRNAs were significantly enriched in Gene Ontology terms such as reproductive processes, cell proliferation, and apoptosis pathways. In addition, an interaction network was constructed by considering target relationships and correlation of the expression levels between ovary development-related genes, miRNAs and pathways. We discovered mRNA and miRNAs transcripts that are candidate regulators of ovary development in egg-ceased geese. Our findings expanded our understanding of the functional of miRNAs in ovarian atrophy and demonstrated that RNA-Seq is a powerful tool for examining the molecular mechanism in regulating egg-ceasing., (Copyright © 2022 Elsevier Inc. All rights reserved.)

1. NOD1 is a significant member of the NOD-like receptor (NLR) family. Its main role is to identify microorganisms that invade the body, transmit immune signals and regulate innate immune responses. However, the expression and role of the NOD1 in immune defence against infection in geese remain unknown.2. The RT-PCR method and rapid amplification of cDNA ends (RACE) was used to obtain the full-length goose NOD1 ( gNOD1 ) cDNA series. The cDNA for gNOD1 contains 2856-bp nucleotides, i.e . 47-bp 5' UTR, 135-bp 3' UTR, and 1275-bp ORF region, and encodes a 951-amino-acids (AAs) polypeptide chain. The nucleotide sequence of gNOD1 was found more than 90% similar to its homologs from other avian organisms.3. The qRT-PCR results showed that gNOD1 mRNA was widely distributed in different tissues, but highly expressed in liver, spleen, lung and caecum tissues.4. Following stimulation of goose embryo fibroblasts (GEFs) with lipopolysaccharide (LPS) and polyriboinosinic polyribocytidylic acid (poly(I:C)), the expression of gNOD1 and cytokines, such as IL-1β, IL-6, IL-18, and TNF-α, changed with the response-efficacy correlation at 24 and 48 h post-infection (hpi).5. When the goslings were challenged with Salmonella entertidis (SE) and LPS, the expression of gNOD1 was up-regulated at 3 and 6 hpi in the spleen and caecum tissues, respectively. However, after SE infection, the expression level of gNOD1 fluctuated, while in the LPS group, gNOD1 mRNA increased immediately at a peak time of 6 hpi and then steadily declined. These results indicated that NOD1 was associated with the potency to resist bacterial and viral infections in the goose, both in vivo and in vitro .

Forkhead box L2 (FOXL2) is a forkhead transcription factor essential for proper reproductive function in females and plays a crucial role in ovarian development in many species of vertebrates. However, little research on goose FOXL2 gene has been conducted. In this study, the cDNA and genomic DNA sequences of goose FOXL2 gene were cloned and characterized. The goose FOXL2 is a single exon gene and contains one open reading frame of 918 bp encoding a protein of 305 amino acids. Bioinformatics analysis displays that the deduced FOXL2 amino acid sequence contains the highly conserved forkhead box domain, which shares greatest similarity to avian species, especially to that of ducks and chicken. RT-qPCR analysis indicates that the FOXL2 mRNA is widely expressed in all examined tissues of fertilized female eggs (28 days), and differentially expressed in female adult (70 days) and laying Zhedong White geese (270 days). Meanwhile, FOXL2 is highly expressed in the hypothalamic-pituitary-ovarian axis, especially in the ovary tissues of the adult and laying geese. Furthermore, one microsatellite (TGTC1415-1418----) and five single nucleotide polymorphisms (A1290G, G1495A, T1554C, T1692A, C1695G and T1697G) were identified in the 3'-untranslated regions. All the information derived from this study could be valuable and facilitate further researches on the function of FOXL2 gene in geese., (Copyright © 2022 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.)