Discover QTLs for the level of blood components in Shaoxing duck using GWAS and haplotype sharing analysis.

Blood composition is indicative of health-related traits such as immunity and metabolism. The use of molecular genetics to investigate alterations in these attributes in laying ducks is a novel approach. Our objective was to employ genome - wide association studies (GWAS) and haplotype - sharing analysis to identify genomic regions and potential genes associated with 11 blood components in Shaoxing ducks. Our findings revealed 35 SNPs and 1 SNP associated with low-density lipoprotein cholesterol (LDL) and globulin (GLB), respectively. We identified 36 putative candidate genes for the LDL trait in close proximity to major QTLs and key loci. Based on their biochemical and physiological properties, TRA2A, NPY, ARHGEF26, DHX36, and AADAC are the strongest putative candidate genes. Through linkage disequilibrium analysis and haplotype sharing analysis, we identified three haplotypes and one haplotype, respectively, that were significantly linked with LDL and GLB. These haplotypes could be selected as potential candidate haplotypes for molecular breeding of Shaoxing ducks. Additionally, we utilized a bootstrap test to verify the reliability of GWAS with small experimental samples. The test can be accessed at https://github.com/xuwenwu24/Bootstrap-test.

Analysis of flavor substances in Shaoxing traditional hand-made Jiafan wine with different amounts of ZaoShao liquor.

BACKGROUND: Adding ZaoShao liquor (high-concentration liquor) is one of the most important steps in the brewing process of Shaoxing Jiafan wine, a product protected by Chinese geographical indications. The focus of this study is the effect of different additive amounts of liquor on the flavor of end products. RESULTS: In this study, four kinds of Shaoxing Jiafan wine were brewed by changing the amount of ZaoShao liquor. Headspace solid-phase microextraction and gas chromatography-mass spectrometry were used to detect the flavor substances of four kinds of Jiafan wine. The difference in flavor of four kinds of Jiafan wine was evaluated by electronic nose analysis technology and verified by sensory evaluation. Finally, the reliability of the experimental results was verified through an aroma reconstruction experiment of rice wine. In this study, the differences in flavoring substances under different amounts of ZaoShao liquor were verified from various angles. The results showed that the flavors of the four kinds of wines were significantly different. CONCLUSION: The composition of flavor substances in Shaoxing rice wine varies with the amount of ZaoShao liquor. This study provided a scientific basis for the improvement of production technology of Shaoxing wine. © 2024 Society of Chemical Industry.

Genome-wide DNA methylation differences between conservation and breeding populations of Shaoxing ducks.

The genome-wide DNA methylation assay was used to analyze the difference in methylation between the breeding and conservation populations of Shaoxing ducks. The methylation level of the breeding population was higher than that of the two conservation populations, and the proportion of CG methylation sites was the largest in the three populations, most of the methylation sites were located in the exon region. There were 1247 different methylation regions in the two populations (group A and B), and 927 different methylation regions in the two groups (group A and group C). The differential methylation regions of the three groups were evenly distributed in the gene and intergene regions. GO and KEGG enrichment analysis showed that the differentially expressed genes in the A and B groups were mainly involved in synaptic and cell connections and the signaling pathways were significantly enriched in cAMP and oxytocin signaling pathways. The results showed that the group C was significantly enriched in eight signaling pathways, including the cAMP signaling pathway and long-term enhancement, compared to the group A. There were thirty-five differentially methylated genes, including CACNA1C, GRIA1, GRIA2, GABBR2, PDE10A, BRAF, GRM5, CPEB3, FMn2, GABRB2, PTK2, and CNTN1. These genes were involved in the development and ovulation of ovaries and follicles and were closely related to the excellent production performance of the breeding population. In addition, ATP2B1, ATP2B2, and other genes related to eggshell quality were identified, which can be used as molecular markers to improve eggshell quality in the future.

Characterization of Bitter Compounds in Shaoxing Huangjiu by Quantitative Measurements, Taste Recombination, and Omission Experiments.

Excessive and uncoordinated bitterness of Shaoxing Huangjiu, a traditional Chinese rice wine, reduces its acceptance by consumers. To determine the compounds responsible for this bitterness, gas chromatography-mass spectrometry and high-performance liquid chromatography were performed on four types of Shaoxing Huangjiu (Yuanhong, Huadiao, Shanniang, and Xiangxue wine) for targeted quantitation of candidate compounds known to contribute to bitterness. Calculations of dose-over-threshold factors revealed that isoamyl alcohol, 1-hexanol, phenylethanol, ethyl butyrate, ethyl lactate, furfural, histidine, and arginine were important bitter compounds. Taste recombination experiments demonstrated that a recombination model constructed using the screened known bitter compounds showed good similarity with the original sample in bitter taste. Furthermore, omission experiments revealed that isobutanol, isoamyl alcohol, 1-hexanol, phenylethanol, ethyl acetate, ethyl butyrate, ethyl lactate, furfural, arginine, and valine were the compounds affecting the bitter taste perception. This study provides a certain guiding effect on the bitterness control and taste improvement of Shaoxing Huangjiu.

Isolation and identification of bitter-tasting peptides in Shaoxing rice wine using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry combined with taste orientation strategy.

The content of peptides in Shaoxing rice wine is high; however, their potential taste properties have not yet been studied. To isolate and identify bitter-tasting peptides in a popular Chinese rice wine, Shaoxing rice wine, a series of purification steps including ultrafiltration and gel filtration chromatography (GFC), combined with a taste orientation strategy, were used. According to the evaluation results of taste dilution analysis, a bitter peptide with the highest taste efficacy was isolated, and its purity was verified by high-performance liquid chromatography (HPLC). The amino acid sequence of peptide components was analyzed by ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS). The bitter peptide Leu-Pro-Thr-Leu (LPTL) was successfully identified from Shaoxing rice wine. Then, taste addition experiments highlighted the bitter impacts of LPTL on sensory descriptors in Shaoxing rice wine. The LPTL peptide can contribute to bitterness in Shaoxing rice wine.

A new luciferase immunoprecipitation system assay provided serological evidence for missed diagnosis of severe fever with thrombocytopenia syndrome.

Severe fever with thrombocytopenia syndrome (SFTS), caused by SFTS virus (SFTSV) infection, was first reported in 2010 in China with an initial fatality of up to 30%. The laboratory confirmation of SFTSV infection in terms of detection of viral RNA or antibody levels is critical for SFTS diagnosis and therapy. In this study, a new luciferase immunoprecipitation system (LIPS) assay based on pREN2 plasmid expressing SFTSV NP gene and tagged with Renilla luciferase (Rluc), was established and used to investigate the levels of antibody responses to SFTSV. Totally 464 serum samples from febrile patients were collected in the hospital of Shaoxing City in Zhejiang Province in 2019. The results showed that 82 of the 464 patients (17.7%) had antibody response to SFTSV, which were further supported by immunofluorescence assays (IFAs). Further, qRT-PCR and microneutralization tests showed that among the 82 positive cases, 15 patients had viremia, 10 patients had neutralizing antibody, and one had both (totally 26 patient). However, none of these patients were diagnosed as SFTS in the hospital probably because of their mild symptoms or subclinical manifestations. All the results indicated that at least the 26 patients having viremia or neutralizing antibody were the missed diagnosis of SFTS cases. The findings suggested the occurrence of SFTS and the SFTS incidence were higher than the reported level in Shaoxing in 2019, and that LIPS may provide an alternative strategy to confirm SFTSV infection in the laboratory.

Immunopotentiators improve the antioxidant defense, apoptosis, and immune response in Shaoxing ducklings.

The abuse of antibiotics for agricultural purposes has been under scrutiny. Therefore, there is an urgent need to find antibiotic substitutes in animal production. The effects of chlorogenic acid, ß-D-Glucan, astragalus flavone, CpG-DNA, and chicken IgG on spleen antioxidant capacity, apoptosis, and the immune response in Shaoxing ducklings were investigated in this study. The ducklings treated with ß-D-Glucan, astragalus flavone, CpG-DNA, and chicken IgG showed significant reduction in catalase and superoxide dismutase activities. The five immunopotentiators facilitated caspase 3 expression and reduced Bcl2 expression in the spleen. Compared to the control group, the protein level of COX2 was significantly upregulated in the chlorogenic acid, CpG-DNA, and chicken IgG groups. The protein level of iNOS expression was significantly improved in all immunopotentiator groups, except for the astragalus flavone group. The five immunopotentiators induced IL-1ß, IFN-α, IFN-ß, TNF-α, RIG-I, TLR3, and TLR7 gene expression. In summary, chlorogenic acid, ß-D-Glucan, astragalus flavone, CpG-DNA, and chicken IgG, as immunopotentiators, improved the innate immune response in the ducklings, which not only provides a new avenue for the development of efficient approaches to prevent pathogen infections, but also offers an alternative to antibiotics in animal production.

Unraveling the difference in aroma characteristics of Huangjiu from Shaoxing region fermented with different brewing water, using descriptive sensory analysis, comprehensive two-dimensional gas chromatography-quadrupole mass spectrometry and multivariate data analysis.

To investigate the specific difference in aroma characteristics of Huangjiu (Chinese rice wine) in Shaoxing region fermented with different brewing water, descriptive sensory analysis, comprehensive two-dimensional gas chromatography-quadrupole mass spectrometry (GC × GC-qMS) and multivariate statistical analysis were employed. The descriptive sensory analysis proved that Huangjiu fermented with Jianhu water had higher overall aroma intensity, and was more prominent in ester, sweet and alcoholic aroma than those fermented with deionized water and Nenjiang water. The results of aroma components analysis by GC × GC-qMS showed that the Huangjiu fermented with Jianhu water had higher concentration of some key aroma compounds, such as ethyl butyrate (OAV: 29-196), isoamyl acetate (OAV: 11-18) and ethyl hexanoate (OAV: 38-47). The multivariate statistical analysis further confirmed that 14 compounds could be used as key markers to distinguish the Huangjiu samples fermented with different brewing water. The correlation network between the volatile compounds in Huangjiu and the inorganic components in water indicated that the ions played an important role in the formation of the difference in aroma characteristics among the samples.

High-temperature exposure alters the community structure and functional features of the intestinal microbiota in Shaoxing ducks (Anas platyrhynchos).

The gut microbiome is a complex ecosystem that contributes to host nutrition and health. However, our current knowledge of the relationship between ambient temperature and gut microbiota of poultry is still limited. The objective of the present study was to characterize the intestinal microbiota of ducks exposed to high ambient temperature. Sixty 60-day-old Shaoxing ducks were allocated to control and heat-treated groups. The ducks in the control group were kept at 25°C, and the ducks in the heat treatment group were raised at 30-40°C, which simulated the temperature change of day and night in summer. After 15 D, the intestinal contents of the duodenum, jejunum, and ileum were obtained from 6 ducks of each group. Genomic DNA was extracted and amplified based on the V4-V5 hypervariable region of 16S rRNA. The results showed that Firmicutes was the dominant bacterial phylum with the highest abundance in the contents of the small intestine of ducks, and the relative abundance of the phylum Firmicutes in all 3 intestinal segments was increased by high temperature. At the genus level, Lactobacillus was found to be the most dominant bacterial genus across 3 gut segments, and its abundance was increased in ducks under heat treatment. Compared with the corresponding intestine segment of control ducks, a total of 36 genera in the duodenum, 19 genera in the jejunum, and 6 genera in the ileum of heat-treated ducks were found to be significantly different in the abundance (linear discriminant analysis score >3.0, P < 0.05). Functional prediction of gut microbiota revealed that high temperature caused changes in the abundance of metabolism and transcription-related pathways. It is noteworthy that most of the altered pathways are related to metabolism. In conclusion, high temperature induced remarkable taxonomic changes in the gut microbiome of ducks, which might be related to the negative effects of high temperature in ducks. Our present study provided an important theoretical ground for high-temperature intervention.

Protective role of dryland rearing on netting floors against mortality through gut microbiota-associated immune performance in Shaoxing ducks.

Dryland rearing on netting floors (DRNF) is a new rearing method for ducks, which could prevent duck excreta from polluting water bodies. However, the influence of DRNF on duck production and immune performance remains poorly understood. In this study, 2,280 Shaoxing ducks, an egg-type breed of Sheldrake in China, were chosen and randomly divided into 2 groups to investigate the effects of DRNF on duck farming. During the experimental laying rates, feed-egg ratios, and mortality rates of the 2 groups were calculated and recorded. Serum immune parameters, including thymus index, spleen index, levels of immunoglobulin G (IgG), interleukin 1ß (IL-1ß), and tumor necrosis factor α (TNF-α), were determined. Sequencing of the 16S rRNA gene was used to analyze the variability of gut microbiota in the duck ileum and cecum. The results showed that DRNF significantly reduced the mortality rate of the ducks and increased the thymus index (P < 0.05), compared to the control. No other significant differences were detected in productional and immune indices (P > 0.05). The 16S rRNA sequencing results revealed differentially enriched microbial compositions in the ileum and cecum, which might be responsible for the improved immune function of Shaoxing ducks. For example, an increase in Lactobacillaceae (family), Anaerotruncus (genus), Saccharibacteria (phylum), Flavobacteriaceae (family), and a reduction in Anaerobiospirillum (genus), Lachnospiraceae (family), Blautia (genus) was revealed in the DRNF ducks. In conclusion, DRNF could alter gut microflora, enhance duck immune system, and reduce mortality in Shaoxing ducks.

Molybdenum and Cadmium exposure influences the concentration of trace elements in the digestive organs of Shaoxing duck (Anas platyrhyncha).

To investigate the toxic effects of Molybdenum (Mo) and Cadmium (Cd) on trace elements in digestive organs of Shaoxing duck (Anas platyrhyncha), 120 Shaoxing ducks were randomly divided into control group and 5 treatment groups which were treated with a commercial diet containing different dosages of Mo and Cd. On the 60th and 120th days, the beak, esophagus, glandular stomach, muscular stomach, small intestine, large intestine and feces were collected to determine contents of Mo, Cd, copper (Cu), iron (Fe), zinc (Zn) and selenium (Se), then correlation analysis was performed. The results showed that Cd content in digestive organs significantly increased in co-treated groups compared to single treated groups and Mo concentration increased in Mo-treated groups compared to control group, whereas Cu, Fe, Zn and Se concentrations in digestive organs decreased in co-treated groups. Furthermore, Cd and Mo were mainly accumulated in the small intestine and esophagus, respectively. There was a strongly positive correlation between Cd and Mo while they had negative correlation with Cu, Fe, Zn and Se, respectively. In feces, Mo and Fe contents in high dose of Mo group and high Mo combined with Cd group were significantly higher than those in control group, and Cu content in all treated groups significantly increased and Cd, Zn and Se concentrations had no difference. The results indicated that dietary Mo or/and Cd might disturb homeostasis of trace elements in digestive organs of Shaoxing duck. Moreover, the two elements presented a synergistic relationship.

Microbial community structure in fermentation process of Shaoxing rice wine by Illumina-based metagenomic sequencing.

BACKGROUND: To understand the role of the community structure of microbes in the environment in the fermentation of Shaoxing rice wine, samples collected from a wine factory were subjected to Illumina-based metagenomic sequencing. RESULTS: De novo assembly of the sequencing reads allowed the characterisation of more than 23 thousand microbial genes derived from 1.7 and 1.88 Gbp of sequences from two samples fermented for 5 and 30 days respectively. The microbial community structure at different fermentation times of Shaoxing rice wine was revealed, showing the different roles of the microbiota in the fermentation process of Shaoxing rice wine. The gene function of both samples was also studied in the COG database, with most genes belonging to category S (function unknown), category E (amino acid transport and metabolism) and unclassified group. CONCLUSION: The results show that both the microbial community structure and gene function composition change greatly at different time points of Shaoxing rice wine fermentation.