Pathogenicity of a goose astrovirus 2 strain causing fatal gout in goslings.

Gosling gout has posed a serious threat to the development of the China's goose industry since the outbreak in mainland China in 2016; goose astrovirus (GAstV) was identified as the culprit pathogen. Two genotypes of this virus have been identified: GAstV-1 and GAstV-2, of which GAstV-2 is the main epidemic strain. Our current understanding of the pathogenic mechanisms of GAstV-2 remains limited. To assess pathogenicity, 1-day-old goslings were inoculated with the GAstV-2 YC20 strain via the subcutaneous, intranasal, and oral infection routes. All the goslings showed typical gout symptoms, with those in the oral infection group exhibiting earlier and more severe clinical symptoms, the highest mortality rate, and greatest weight loss. The blood biochemical indicators, viral loads in cloacal swabs and all representative tissues, and serum antibody titers of all infection groups increased significantly, and no significant differences in these parameters were observed among the three infection groups. Histopathological studies showed that the livers, kidneys, and spleens were the main damaged organs, and the pathological changes in the oral group were more severe than those in the other groups. Further analysis revealed that hepatic sinuses narrowed or became occluded as early as 1 day post-inoculation; urate deposition occurred in the renal tubules at 2 days post-inoculation (dpi), followed by necrosis of renal tubular epithelial cells; and lymphocytic infiltration appeared in the splenic tissue at 5 dpi. These results further our understanding of the pathogenic mechanisms of GAstV-2 and provide a reference for future studies.

Host innate immune responses of geese infected with goose origin nephrotic astrovirus.

A novel goose astrovirus (GoAstV) outbreak in goslings, characterized by severe articular and visceral gout with high mortality, occurred in China. Although the pathogenesis of GoAstV-infected goslings has been explored in several studies, the host-immune response remains unclear. In this study, a goose astrovirus was isolated from goslings in Xiaogan, and designated as the HBXG strain. The full-length genome of HBXG was 7170 nt. A sequence analysis and phylogenetic trees revealed HBXG belonged to the novel GoAstV. We evaluated the viral distribution systematically and estimated immune related gene expression in HBXG-infected goslings. Results showed that GoAstV replicated quickly in many tissues and the highest titer was observed in the kidney, which reached 109.6 copies. TLR3, RIG-I and MDA5 were involved in the host-immune response to GoAstV, and the expression of IFN types I (IFN-α, IFN-ß), inflammatory cytokines (IL-8, IL-10, TNF-α), antiviral proteins (Mx, OASL, PKR) and MHC-I were also upregulated during the infection. In contrast, the expression of proinflammatory cytokines (IL-1ß, IL-6) and MHC-II were inhibited at 3 dpi. This study suggests that GoAstV is highly pathogenic to goslings, causing multiple systemic infections in tissues and the host-immune response is activated early in infection. However, rapid viral replication, suppression of inflammatory cytokines (IL-1ß, IL-6) and MHC-II expressions were the possible reasons why the host-immune response cannot provide enough protection against GoAstV infection. This study is the first report to illuminate the immune response in goslings infected with GoAstV and offers insight into the pathogenesis of GoAstV.

Effects of astragalus polysaccharide on intestinal inflammatory damage in goslings infected with gosling plague.

1. This study explored the effects of Astragalus membranaceus polysaccharide (APS) on intestinal inflammatory damage of goslings infected with parvovirus ('gosling plague').2. A total of 90 healthy goslings were randomly divided into three groups; control, infected or APS treated, respectively. Goslings in the infection and APS treatment groups were inoculated with 0.3 ml allantoic fluid containing goose parvovirus (ELD50 = 1 × 103/0.3 ml) by intramuscular injection and the control group were injected with saline (0.3 ml) twice a day for 15 days.3. Blood serum and the jejunum were collected at 5, 10 and 15 days after the start of the experiment to detect the activities of SOD and GSH-Px, levels of MDA, sIgA, IL-1ß, IL-6 and TNF-α, the mRNA expression of IL-1ß, IL-6, LITAF, NF-κB, COX-2 and PGE2, pathological damage in the jejunum and serum IgG, IgM, C3, C4, IFN-γ levels.4. After APS treatment, SOD and GSH-Px activities increased, MDA content decreased; sIgA, IL-1ß, IL-6 and TNF-α protein content, and IL-1ß, IL-6, LITAF, NF-κB, COX-2 and PGE2 mRNA expression decreased in the jejunal tissue, serum IgG, IgM, C3, C4, IFN-γ significantly increased and pathological damage of jejunum significantly improved.5. In conclusion, APS reduced intestinal inflammatory damage in goslings infected with parvovirus by improving the immune and antioxidant functions of goslings.

The endogenous development and pathogenicity of Eimeria anseris (Kotlan, 1932) in domestic goslings.

Twenty-one, 25-day-old, artificially reared, coccidia-free goslings (Anser cygnoides var. domestica) were inoculated orally with 0.5 × 104, 1 × 104, or 100 × 104 sporulated oocysts of Eimeria anseris and sacrificed at intervals from 24 to 216 h post-inoculation (HPI). Nine uninfected goslings served as negative controls. Parts of the visceral organs from goslings, including the intestines, kidneys, and liver, were fixed, sectioned, and observed microscopically. The results revealed that two generations of meronts occurred in the life cycle of E. anseris. The first generation of meronts developed at 24-96 HPI and the second generation at 90-128 HPI. Each meront contained 4-10 merozoites. Development of gamonts began at 128 HPI and mature oocysts appeared at 168 HPI. Developmental stages presented mainly in the epithelial cells of crypts and lamina propria in the posterior parts of the jejunum and ileum. Parasites localized mostly in the cytoplasm and occasionally in the nuclei of host cells. Histological lesions were pronounced in the jejunum and ileum. Desquamation and necrosis of the epithelium of intestine and crypts, infiltration of inflammatory cells, and hemorrhage and mucosal edema were associated with aggregates of endogenous stages. The infected goslings mainly showed severe diarrhea, depression, anorexia, and emaciation, suggesting that E. anseris is highly pathogenic in goslings.

Immunobiological activity and antiviral regulation efforts of Chinese goose (Anser cygnoides) CD8α during NGVEV and GPV infection.

The CD8 molecule is a cell membrane glycoprotein expressed on cytotoxic T lymphocytes, which are involved in the clearance of viruses. However, the functional characterization of goose CD8α is still unclear. The immunobiological characterization of goose CD8α in goose spleen mononuclear cells (MNCs) was examined by real-time quantitative PCR (qPCR). It was shown that CD8α mRNA levels were significantly up-regulated by in vitro treatment of MNCs with phytohemagglutinin (PHA), concanavalin A (ConA), and polyinosinic-polycytidylic acid (poly I:C) in a dose-dependent way, but lipopolysaccharides (LPSs) did not have this same effect. Moreover, the time-course effect of CD8α expression in response to mitogens (PHA, ConA, and poly I:C) was evaluated in MNCs. A significant increase in the transcriptional levels of CD8α was detected in new type gosling viral enteritis virus (NGVEV)-infected goose MNCs at 48 h postinfection (PI) and in goose parvovirus (GPV)-infected MNCs at 72 h PI. Also, the number of CD8α+ cells was significantly increased during viral infection from 72 h on. The seminal changes in mRNA profiles of antiviral cytokines (IFN-α, IFN-γ, and IL-18) were observed and were significantly increased during late phases of NGVEV and GPV infection. Accordingly, our data not only contribute to the understanding of the immune characteristics of goose CD8α, but they also provide new insight into the innate antiviral immunity of geese.

Transcriptome Analysis and Identification of Differentially Expressed Transcripts of Immune-Related Genes in Spleen of Gosling and Adult Goose.

The goose (Anser cygnoides), having high nutritional value, high-quality feathers and high economic benefit, is an economically important poultry species. However, the molecular mechanisms underlying the higher susceptibility to pathogens in goslings than in adult geese remains poorly understood. In this study, the histological sections of spleen tissue from a two-week-old gosling and an adult goose, respectively, were subjected to comparative analysis. The spleen of gosling was mainly composed of mesenchyma, accompanied by scattered lymphocytes, whereas the spleen parenchyma was well developed in the adult goose. To investigate goose immune-related genes, we performed deep transcriptome and gene expression analyses of the spleen samples using paired-end sequencing technology (Illumina). In total, 50,390 unigenes were assembled using Trinity software and TGICL software. Moreover, these assembled unigenes were annotated with gene descriptions and gene ontology (GO) analysis was performed. Through Kyoto encyclopedia of genes and genomes (KEGG) analysis, we investigated 558 important immune-relevant unigenes and 23 predicted cytokines. In addition, 22 immune-related genes with differential expression between gosling and adult goose were identified, among which the three genes showing largest differences in expression were immunoglobulin alpha heavy chain (IgH), mannan-binding lectin serine protease 1 isoform X1 (MASP1) and C-X-C chemokine receptor type 4 (CXCR4). Finally, of these 22 differentially expressed immune-related genes, seven genes, including tumor necrosis factor receptor superfamily member 13B (TNFRSF13B), C-C motif chemokine 4-like (CCL4), CXCR4, interleukin 2 receptor alpha (IL2RA), MHC class I heavy chain (MHCIα), transporter of antigen processing 2 (TAP2) IgH, were confirmed by quantitative real-time PCR (qRT-PCR). The expression levels of all the candidate unigenes were up-regulated in adult geese other than that of TNFRSF13B. The comparative analysis of the spleen transcriptomes of gosling and adult goose may promote better understanding of immune molecular development in goose.

Effects of dietary supplementation of vitamin A on the tibia quality of goslings.

OBJECTIVE: This study was conducted to evaluate the effect of dietary supplementation of vitamin A (VA) on the tibial growth, calcium (Ca) and phosphorus (P) metabolism, VA, and vitamin D (VD) deposition, and associated gene expression in goslings. METHODS: A total of 180 healthy, 1-day-old male goslings were randomly divided into 3 treatment groups (0, 9,000, and 15,000 IU VA/kg), with 6 replicates containing 10 goslings each. They were weighed and sampled on days 14, 28, 42, 56, and 70. RESULTS: No addition of VA reduced VA content in the serum and liver of goslings, and supplementation of 15,000 IU/kg VA increased VA content from day 14 (p<0.05). The trend of VA concentration in the serum and liver was in line with the relative mRNA expression of retinoic acid receptor ß in the jejunal mucosa. In both no addition of VA and supplementation of 15,000 IU/kg VA reduced 25-hydroxycholecalciferol (25-OH-VD3) content in the serum and VD content in the liver (p<0.05). From day 28, no addition of VA or supplementation of 15,000 IU/kg VA had a negative effect on tibia length, strength, and Ca, P, and ash content in goslings (p<0.05). Tibia P content was lower in the supplementation of 15,000 IU/kg VA group than in the no addition of VA group (p<0.05). No addition of VA or supplementation of 15,000 IU/kg VA had the most effect on early serum parathyroid hormone (PTH) levels in goslings (p<0.05). The effect of no addition of VA on the bone Gla protein (BGP) content of goslings started from day 14 (p<0.05). The relative mRNA expression of bone Gla-protein (BGLAP) and bone morphogenetic protein 4 (BMP4) in the liver and jejunal mucosa was decreased by either no addition of VA or supplementation of 15,000 IU/kg VA (p<0.05). CONCLUSION: Both no addition of VA and supplementation of 15,000 IU/kg VA affected the mineralization process of the bone, and ultimately reduced tibial quality.

Aminoguanidine alleviates gout in goslings experimentally infected with goose astrovirus-2 by reducing kidney lesions.

Goose astrovirus (GAstV)-2, a novel pathogen identified in 2018, mainly causes visceral gout in goslings, leading to approximately 50% mortality. At present, no commercial veterinary products are available to prevent and treat the disease. Our previous studies showed that nitric oxide (NO) and inducible NO synthase (iNOS) were markedly higher in the kidney and spleen of goslings infected with GAstV-2, but their effects during GAstV-2 infection remain unclear. In the present study, goslings were intraperitoneally injected with aminoguanidine (AG)-an iNOS inhibitor-to examine the role of NO during GAstV-2 infection. AG significantly decreased the serum NO concentration and iNOS mRNA expression in the kidney. Moreover, AG reduced the mortality, serum uric acid and creatinine content, and urate deposition in visceral organs and joints. Histopathological analysis demonstrated that AG reduced renal tubular cell necrosis, inflammatory cell infiltration, glycogen deposition in glomerular mesangium, and interstitial fibrosis, suggesting alleviation of kidney lesions. Furthermore, AG decreased the expression of renal injury markers such as KIM-1 and desmin; inflammatory cytokine-related genes such as IL-1ß, IL-8, and MMP-9; and autophagy-related genes and proteins such as LC3II, ATG5, and Beclin1. However, quantitative real-time PCR and immunohistochemistry showed that treatment with AG did not affect the kidney and liver viral load. These findings suggest that AG decreases the mortality rate and kidney lesions in goslings infected with GAstV-2 through mechanisms associated with autophagy and inhibition of inflammatory cytokine production in the kidney but not with GAstV-2 replication.

Neonatal Care of Anseriformes.

Neonatal Anseriformes require specialized care for successful development including access to clean swimming water and food presentation that stimulates natural feeding behavior. Knowledge of natural history is essential for successful rearing. Lightweight and waterproof materials can be used for corrective splinting of many developmental disorders that allow birds to ambulate normally and swim. Captivity-associated lesions can be minimized through proper husbandry and hygiene. Anseriformes are susceptible to a variety of bacterial, viral, fungal, and parasitic diseases, some of which are reportable to state health agencies.

In ovo betaine injection improves breast muscle growth in newly hatched goslings through FXR/IGF-2 pathway.

Betaine has been shown to enhance growth performance and increase breast muscle yield in ducks and broilers through various mechanisms, including the modification of DNA methylation. However, the impact of in ovo betaine injection on muscle growth in newly hatched goslings remains unclear. In this study, fifty eggs were injected with saline or betaine at 7.5 mg/egg prior to incubation, and the subsequent effects on breast muscle growth in the newly hatched goslings were investigated. Betaine significantly increased (P < 0.05) the hatch weight, breast muscle weight, and breast muscle index, accompanied by an augmentation in muscle bundle cross-sectional area. Concurrently, betaine significantly upregulated (P < 0.05) the expression levels of myogenic regulatory factors, including myogenin (MyoG) and paired box 7 (Pax7) both mRNA and protein, while downregulating (P < 0.05) the mRNA and protein levels of myostatin (MSTN). Histological analysis revealed a higher abundance of proliferating cell nuclear antigen (PCNA) and Pax7 immune-positive cells in the breast muscle of the betaine group, consistent with elevated PCNA and Pax7 mRNA and protein levels. Additionally, significantly increased (P < 0.05) contents of insulin-like growth factor 1 (IGF-1) and insulin-like growth factor 2 (IGF-2) were observed in the breast muscle of the betaine group, so was mRNA expression of IGF-1, IGF-2, and insulin-like growth factor 1 receptor (IGF-1R). Betaine also significantly in8creased (P < 0.05) global DNA methylation of the breast muscle, accompanied by enhanced mRNA and protein levels of methionine cycle and DNA methylation-related enzymes, Interestingly, the promoter regions of IGF-1, IGF-2, and IGF-1R genes were significantly hypomethylated (P < 0.05). Moreover, in ovo betaine injection significantly upregulated (P < 0.05) the protein level of farnesoid X receptor (FXR) in breast muscle and FXR binding to the promoter of IGF-2 gene. These findings suggest that in ovo betaine injection promotes breast muscle growth during embryonic development in goslings through the FXR-mediated IGF-2 pathway, ultimately improving hatch weight and breast muscle weight.

Tibial Damage Caused by T-2 Toxin in Goslings: Bone Dysplasia, Poor Bone Quality, Hindered Chondrocyte Differentiation, and Imbalanced Bone Metabolism.

T-2 toxin, the most toxic type A trichothecene, is widely present in grain and animal feed, causing growth retardation and tissue damage in poultry. Geese are more sensitive to T-2 toxin than chickens and ducks. Although T-2 toxin has been reported to cause tibial growth plate (TGP) chondrodysplasia in chickens, tibial damage caused by T-2 toxin in geese has not been fully demonstrated. This study aims to investigate the adverse effects of T-2 toxin on tibial bone development, bone quality, chondrocyte differentiation, and bone metabolism. Here, forty-eight one-day-old male Yangzhou goslings were randomly divided into four groups and daily gavaged with T-2 toxin at concentrations of 0, 0.5, 1.0, and 2.0 mg/kg body weight for 21 days, respectively. The development of gosling body weight and size was determined by weighing and taking body measurements after exposure to different concentrations of T-2 toxin. Changes in tibial development and bone characteristics were determined by radiographic examination, phenotypic measurements, and bone quality and composition analyses. Chondrocyte differentiation in TGP and bone metabolism was characterized by cell morphology, tissue gene-specific expression, and serum marker levels. Results showed that T-2 toxin treatment resulted in a lower weight, volume, length, middle width, and middle circumference of the tibia in a dose-dependent manner (p < 0.05). Moreover, decreased bone-breaking strength, bone mineral density, and contents of ash, Ca, and P in the tibia were observed in T-2 toxin-challenged goslings (p < 0.05). In addition, T-2 toxin not only reduced TGP height (p < 0.05) but also induced TGP chondrocytes to be disorganized with reduced numbers and indistinct borders. As expected, the apoptosis-related genes (CASP9 and CASP3) were significantly up-regulated in chondrocytes challenged by T-2 toxin with a dose dependence, while cell differentiation and maturation-related genes (BMP6, BMP7, SOX9, and RUNX2) were down-regulated (p < 0.05). Considering bone metabolism, T-2 toxin dose-dependently and significantly induced a decreased number of osteoblasts and an increased number of osteoclasts in the tibia, with inhibited patterns of osteogenesis-related genes and enzymes and increased patterns of osteoclast-related genes and enzymes (p < 0.05). Similarly, the serum Ca and P concentrations and parathyroid hormone, calcitonin, and 1, 25-dihydroxycholecalciferol levels decreased under T-2 toxin exposure (p < 0.05). In summary, 2.0 mg/kg T-2 toxin significantly inhibited tibia weight, length, width, and circumference, as well as decreased bone-breaking strength, density, and composition (ash, calcium, and phosphorus) in 21-day-old goslings compared to the control and lower dose groups. Chondrocyte differentiation in TGP was delayed by 2.0 mg/kg T-2 toxin owing to cell apoptosis. In addition, 2.0 mg/kg T-2 toxin promoted bone resorption and inhibited osteogenesis in cellular morphology, gene expression, and hormonal modulation patterns. Thus, T-2 toxin significantly inhibited tibial growth and development with a dose dependence, accompanied by decreased bone geometry parameters and properties, hindered chondrocyte differentiation, and imbalanced bone metabolism.

Rapid and visual detection of an isolated and identified goose parvovirus (GPV) strain by a loop-mediated isothermal amplification assay.

Gosling plague caused by goose parvovirus (GPV), a highly infectious septic disease with high mortality, has caused substantial loss in the waterfowl industry. A method for the rapid detection of GPV is needed. In this study, we isolated the virus strain of GPV in May 2020 and applied it to the loop-mediated isothermal amplification (LAMP) assay. We designed five sets of primers for the goose parvovirus VP3 gene by LAMP. The GV-1 primer set was selected to detect GPV sensitively and rapidly. LAMP was more sensitive compared to PCR. In addition, the LAMP method could complete detection within 60 min which was faster than the PCR assay. The LAMP provided a convenient and effective experimental method for detection of GPV for inspection and quarantine departments and health care units in China, and it is expected to become a simple and routine detection method, especially suitable for goose farms.

Egg quality, hatchability, gosling quality, and amino acid profile in albumen and newly-hatched goslings' serum as affected by egg storage.

In modern poultry husbandry, storing fertilized eggs is a common measure to cope with the variable demands of the market and the maximum hatching capacity of the hatchery. However, this measure is harmful to the hatchability of eggs and the quality of newly hatched birds. Knowledge about the effects of storing fertilized eggs on the performance of goslings is still limited. The objective of this study was to investigate the effects of storing fertilized eggs on egg quality, hatchability, gosling quality, hatching weight, post-hatching growth performance, and amino acid profile in albumen and newly hatched goslings' serum. A total of 1,080 fertilized goose eggs (Jilin White goose) with a similar egg weight (126.56 ± 0.66 g) were used in this study. All eggs were distributed into 3 groups with 24 replicates per group and 15 eggs per replicate. The differences between groups were the storage duration of eggs (0, 7, or 14 d). We found that the Haugh unit, yolk weight, and eggshell weight decreased linearly, whereas the albumen pH increased linearly, with storage duration. Prolonging storage duration had negative effects on hatchability, hatching weight, post-hatching growth performance parameters, and gosling quality in a time-dependent manner. The analysis of the amino acid profile in albumen and newly-hatched goslings' serum showed that the amino acid content increased linearly with storage duration. Additionally, eggs stored for 14 d had the worst performance for all measured parameters. Therefore, we concluded that the storage of fertilized eggs negatively affects egg quality and post-hatching gosling quality. To produce high-quality goslings, it is necessary to shorten the storage duration for fertilized eggs.

Bacterial lipopolysaccharide with different administration routes affects intestinal mucosal morphological, immunological, and microbial barrier functions in goslings.

The current study was conducted to evaluate the effects of different administration routes of bacterial lipopolysaccharide (LPS) on intestinal mucosal morphological, immunological, and microbial barrier functions in goslings. First, we compared intestinal villi morphology of goslings under intraperitoneal or oral LPS treatment through hematoxylin and eosin staining. Then, we determined the signatures of the microbiome in the ileum mucosa of goslings subjected to oral LPS treatment at 0, 2, 4, and 8 mg/kg BW by 16S sequencing, and analyzed the changes in intestinal barrier functions and permeability, levels of LPS in the ileum mucosa, plasma, and liver tissue, and the induced inflammatory response of Toll-like receptor 4 (TLR4). As a result, intraperitoneal LPS injection resulted in a thicker intestinal wall in the ileum within a short time, whereas villus height was less affected; in contrast, oral LPS treatment exerted a stronger influence on villus height but not on intestinal wall thickness. We also found that oral LPS treatment affected the structure of the intestinal microbiome, reflected by changes in the clustering of intestinal microbiota. The average abundance of Muribaculaceae showed an increasing trend with increasing LPS levels, and that of the genus Bacteroides decreased, compared with the control group. In addition, oral LPS treatment with 8 mg/kg BW affected the intestinal epithelial morphology, damage the mucosal immune barrier, downregulated the expression of tight junction proteins, increased circulating D-lactate levels, and stimulated the secretion of various inflammatory mediators and activation of the TLR4/MyD88/NFκB pathway. This study presented the injuries of intestinal mucosal barrier function induced by LPS challenges in goslings and provided a scientific model for searching the novel strategies to attenuate the immunological stress and gut injury caused by LPS.

Polysaccharide of Atractylodes macrocephala Koidz alleviate lipopolysaccharide-induced liver injury in goslings via the p53 and FOXO pathways.

Lipopolysaccharide (LPS) can affect the immune system of geese by inducing liver injury. The polysaccharide of Atractylodes macrocephala Koidz (PAMK) have obvious immune-enhancing effects. Accordingly, this experiment investigated the effect of PAMK on LPS-induced liver injury in goslings. Two hundred 1-day-old goslings were randomly divided into the control group, LPS group, PAMK group, and PAMK+ LPS group, and the PAMK and PAMK+ LPS groups were fed the basal diet with 400 mg/kg PAMK, while the control and LPS groups were fed the basal diet. On D 21, 23, and 25 of the formal trial, the goslings in the LPS and PAMK+LPS groups were injected intraperitoneally with 2 mg/kg LPS, and goslings in the control and PAMK groups were injected intraperitoneally with the same amount of saline. Livers were collected on D 25. HE-stained sections showed that PAMK could effectively alleviate the LPS-induced indistinct hepatic cord structure, loss of cytoplasmic contents of hepatocytes, and dilatation of hepatic sinusoids. The biochemical parameters of liver tissues showed that PAMK could alleviate the LPS-induced upregulation of alanine aminotransferase and aspartate aminotransferase. To further investigate the mechanism of the mitigating effect of PAMK on LPS-induced injury, livers from the LPS and PAMK+LPS groups were selected for transcriptome sequencing. The sequencing results showed that there were 406 differentially expressed genes (DEGs) in the livers of LPS and PAMK+LPS goslings, of which 242 upregulated and 164 downregulated. The Kyoto Encyclopedia of Genes and Genome (KEGG) analysis showed that DEGs were significantly enriched in immune signal transduction, cell cycle, and cell metabolism. Besides, protein‒protein interaction analysis showed that 129 DEGs were associated with each other, including 7 DEGs enriched in the p53 and FOXO signaling pathway. In conclusion, PAMK may alleviate LPS-induced liver injury in gosling through the p53 and FOXO signaling pathway. These results provide a basis for further development of PAMK as an immunomodulator.

Development of small intestine and sugar absorptive capacity in goslings during pre- and post-hatching periods.

This study was conducted to investigate the development patterns of small intestine, intestinal morphology, disaccharidase activities, and sugar transporter gene expression in goslings during pre- and post-hatching periods. Small intestine was sampled on embryonic d 23 and 27, day of hatch, and d 1, 4, and 7 post-hatching. A total of 18 eggs with the breed of Jilin White geese were selected at each sampling timepoint for measuring relevant parameters. Three eggs were considered as a group, with 6 groups in each sampling timepoint. Rapid development of small intestine was observed around the hatching, of which jejunum and ileum had relatively higher development rates. Villus surface area from three intestinal segments started to increase on embryonic d 27, and kept relatively stable during day of hatch to d 1 post-hatching, and following increased till d 7 post-hatching. A high priority of villi enrichment was observed in duodenum and jejunum. The activity of disaccharidase increased before hatching and kept relatively high-level post-hatching, of which the activity of disaccharidase was highest in jejunum. The expression of sugar transporter gene increased prior to hatching and then decreased post-hatching, of which jejunum and duodenum were sites with high sugar transporter gene expression. Rapid development in intestinal morphology, disaccharidase activities, and sugar transporter gene expression around the hatching indicated that goslings have high potential to digest and/or assimilate carbohydrates during its early-life, which provided a preparation for further digestion of exogenous feed. This study provided a profile of development patterns for intestinal morphology, disaccharidase activities, and sugar transporter gene expression in goslings, which was beneficial to understanding the characteristics of nutrient absorption during the early-life of goslings.

Polysaccharide of Atractylodes macrocephala Koidz alleviate lipopolysaccharide-stimulated liver inflammation injury of goslings through miR-223/NLRP3 axis.

Lipopolysaccharide (LPS) infection could cause severe liver inflammation and lead to liver damage, even death. Previous studies have shown that polysaccharide of Atractylodes macrocephala Koidz (PAMK) could protect liver from inflammation caused by LPS in mice. However, whether PAMK could alleviate liver inflammatory injury in other animals with LPS is still unknown. For evaluating whether PAMK could alleviate liver inflammatory injury in goslings with LPS, a total of 80 healthy 1-day old Magang goslings were randomly divided into 4 groups (control group, PAMK group, LPS group, and PAMK+LPS group). Goslings in control group and LPS group were fed with basal diet, and goslings in PAMK group and PAMK+LPS group were fed basal diet supplemented with 400 mg/kg PAMK to the end of trial. On 24 d of age, goslings in the control group and PAMK group were intraperitoneal injected 0.5 mL normal saline, and goslings in LPS and PAMK+LPS groups were intraperitoneal injected with LPS at 5 mg/kg BW. The serum and liver samples were collected for further analysis after treatment of LPS at 6, 12, 24, and 48 h. Furthermore, the hepatocytes were extracted from goose embryo to measure the expression of the key genes of miR-223/NLRP3 axis. The results showed that PAMK pretreatment could maintain normal cell morphology of liver, alleviate the enhanced levels of biochemical indexes ALT and AST, decrease the levels of IL-1ß and IL-18, increase the relative mRNA expression of miR-223, and decrease the expression of NLRP3, Caspase-1, and cleaved Caspase-1 in liver and hepatocytes of goslings induced by LPS. These results indicated that PAMK could relieve inflammatory liver tissue damage after LPS treatment and downregulate the level of inflammation factors via miR-223/NLRP3 axis, thus playing a liver protective role in liver inflammation injury in goslings.

Extensive genetic heterogeneity and molecular characteristics of emerging astroviruses causing fatal gout in goslings.

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.

Development of breast muscle parameters, glycogen reserves, and myogenic gene expression in goslings during pre- and post-hatching periods.

This study aimed to better understand the development patterns of breast muscle and glycogen reserves in goslings during pre- and post-hatching periods. The timepoints for sampling were embryonic days 23 and 27 of hatching and days 1, 4, and 7 post hatching. We found that the body weight of goslings increased with age. The small intestine developed with age and remained reasonably constant on day 4 post hatching. The breast muscle development decreased with age and stayed relatively stable on day 1 post hatching. The diameter of myofiber increased prior to hatching and then decreased while hatching. The development patterns of breast muscle glycogen reserves were similar to the diameter of myofiber. In contrast, the contents of liver glycogen began to decrease before hatching and then increased rapidly after hatching. Moreover, the expression of Myf-5 increased with age. The expression of MSTN was maintained at high levels prior to hatching, dropped immediately after hatching, and then gradually increased with age. Additionally, we also observed that the glycogen content in the breast muscle was positively correlated with the diameter of the myofiber. The liver glycogen content was positively correlated to the relative weight of the breast muscle, the diameter of the myofiber, and the breast muscle glycogen content. The development pattern of the myofiber was synchronized with the change in the MSTN/Myf-5 ratio. This study provided a profile to understand the development patterns of breast muscle, glycogen reserves, and myogenic gene expression in goslings, which was beneficial to understanding the characteristics of energy reserves during the early life of goslings.

Brooding Temperature Alters Yolk Sac Absorption and Affected Ovarian Development in Goslings.

In order to explore the brooding temperature on the absorption of yolk sac and the ovary development of goslings, 126 1-day-old female goslings were randomly divided into three groups with three replicates in each group. The brooding temperatures were set at 32 °C, 29 °C and 26 °C (represent G32, G29 and G26), respectively, in each group. At 48, 60 and 72 h, two goslings from each replicate were weighed, and the yolk sac was collected and weighed. The fatty acid composition of yolk sac fluid was determined by gas chromatography-mass spectrometry (GC-MS). At 1, 2, 3, and 4 weeks of age, goslings from each replicate were weighed, the ovaries were weighed and fixed for hematoxylin-eosin (HE) staining, Cell cycle checkpoint kinase 1 (CHK1), fibroblast growth factor 12 (FGF12) and Sma-and Mad-related protein 4 (SMAD4) which related to regulation of ovarian development were determined by qRT-PCR. The body weight of G29 and G26 was significantly higher than that of G32 at 72 h (p < 0.05). The contents of C14:0, C16:0, C18:2n6c and total fatty acid (ΣTFA) from G32 were significantly higher than that of G26 (p < 0.05), and the contents of C18:1n9t and C22:0 in G29 were significantly higher than that of G26 (p < 0.05). The ovary index, ovary and body weight were significantly higher in G29 than those of G32 and G26 at 2 weeks of age (p < 0.05). The number of primordial follicles, number of primary follicles and diameter of primary follicles were significantly higher in G29 than those in G32 and G26 at 4 weeks of age (p < 0.05). In G29, the expression of CHK1 and SMAD4 was significantly higher than that in G32, and the expression of FGF12 and SMAD4 was significantly higher (p < 0.05) than that in G26 at 2 and 4 weeks of age. In conclusion, brooding temperature at 29 °C could promote the absorption of fatty acids in yolk sac, body weight gain, and ovarian development through up-regulating the expression of CHK1, FGF12 and SMAD4.