摘要：
Spirulina ( Arthrospira ) has been extensively applied in CO 2 biofixation, wastewater purification, and value-added bioproducts preparation. Light availability plays a pivotal role in Spirulina photoautotrophic cultivation, which is primary determined by characteristics of incident light and distribution of light within photobioreactors (PBRs). To clarify the role of light in Spirulina photoautotrophic cultivation, this review first analyzes the processes of light delivery and conversion in suspended PBRs. Then, effects of key light characteristics, including light intensity, spectrum, and photoperiod, on Spirulina growth and intracellular biochemical components synthesis are comprehensively summarized. Recent advancements in innovative PBR designs aimed at enhancing light utilization efficiency and promoting Spirulina growth are also highlighted. Finally, potential future research directions in the field of Spirulina photoautotrophic cultivation are outlined. Overall, this work provides a theoretical foundation and technical guidance for improving Spirulina production and specific target products synthesis from prespectives of light conditions regulation and PBRs design.
Spirulina ( Arthrospira ) has been extensively applied in CO 2 biofixation, wastewater purification, and value-added bioproducts preparation. Light availability plays a pivotal role in Spirulina photoautotrophic cultivation, which is primary determined by characteristics of incident light and distribution of light within photobioreactors (PBRs). To clarify the role of light in Spirulina photoautotrophic cultivation, this review first analyzes the processes of light delivery and conversion in suspended PBRs. Then, effects of key light characteristics, including light intensity, spectrum, and photoperiod, on Spirulina growth and intracellular biochemical components synthesis are comprehensively summarized. Recent advancements in innovative PBR designs aimed at enhancing light utilization efficiency and promoting Spirulina growth are also highlighted. Finally, potential future research directions in the field of Spirulina photoautotrophic cultivation are outlined. Overall, this work provides a theoretical foundation and technical guidance for improving Spirulina production and specific target products synthesis from prespectives of light conditions regulation and PBRs design.
作者机构：
[Liu, Zhi-Wei; Tang, Pan-Pan; Tan, Yi-Cheng; Liu, Tian-Le] College of Food Science and Technology, Hunan Agricultural University, Changsha, 410128, China;[Liu, Xiu-Bin] College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, China;[Liu, Chang; Cheng, Jun-Hu] School of Food Science and Engineering, South China University of Technology, Guangzhou, 510641, China;[Aadil, Rana Muhammad] National Institute of Food Science and Technology, University of Agriculture, Faisalabad, 38000, Pakistan
通讯机构：
[Zhi-Wei Liu] C;College of Food Science and Technology, Hunan Agricultural University, Changsha, 410128, China
摘要：
The potential of cold plasma (CP) treatment to promote the formation of amyloid fibrils (AFs) of ovalbumin (OVA) was evaluated relative to acidic heat fibrillation condition (pH = 2, 85 °C). Results indicated CP exhibited significant potential for promoting the formation of AFs of OVA, as evidenced by higher Thioflavin T (ThT) fluorescence intensity of AFs for CP-treated OVA (POVA) during the fibrillation process compared to AFs for native OVA under acidic heating conditions (NOVA). Long, curved and worm-like fibrils of CP-treated OVA with 2 min (POVA-2), which were thicker (3 nm in height) and longer (majority length ranging from 300 nm to 400 nm), were observed after 8 h of fibrillation, while irregular, short, worm-like fibrils of NOVA were detected. Unlike NOVA, which was only hydrolyzed into small peptides, the backbone of POVA-2 was cleaved into small peptide fragments, accompanied by the generation of dityrosine cross-linked aggregation/oligomer during fibrillation process, as evidenced by SDS-PAGE and dityrosine analysis. The result of hydrophobic, sulfhydryl and disulfide bonds, and dityrosine analysis showed hydrophobic interaction and dityrosine cross-links could be the main force driving the assembly and stacking of cross-β structures, leading to the formation of organized fibrillar structures for POVA-2, while only hydrophobic interaction was involved for NOVA. Additionally, analyses of emulsifying ability (EAI) and stability (ESI) of NOVA and POVA-2 displayed that both EAI and ESI of them were significantly improved, with POVA-2 exhibiting superior EAI compared to NOVA. Therefore, this study demonstrated CP is a promising technique to promote the generation of protein amyloid fibrils in a more efficient manner.
The potential of cold plasma (CP) treatment to promote the formation of amyloid fibrils (AFs) of ovalbumin (OVA) was evaluated relative to acidic heat fibrillation condition (pH = 2, 85 °C). Results indicated CP exhibited significant potential for promoting the formation of AFs of OVA, as evidenced by higher Thioflavin T (ThT) fluorescence intensity of AFs for CP-treated OVA (POVA) during the fibrillation process compared to AFs for native OVA under acidic heating conditions (NOVA). Long, curved and worm-like fibrils of CP-treated OVA with 2 min (POVA-2), which were thicker (3 nm in height) and longer (majority length ranging from 300 nm to 400 nm), were observed after 8 h of fibrillation, while irregular, short, worm-like fibrils of NOVA were detected. Unlike NOVA, which was only hydrolyzed into small peptides, the backbone of POVA-2 was cleaved into small peptide fragments, accompanied by the generation of dityrosine cross-linked aggregation/oligomer during fibrillation process, as evidenced by SDS-PAGE and dityrosine analysis. The result of hydrophobic, sulfhydryl and disulfide bonds, and dityrosine analysis showed hydrophobic interaction and dityrosine cross-links could be the main force driving the assembly and stacking of cross-β structures, leading to the formation of organized fibrillar structures for POVA-2, while only hydrophobic interaction was involved for NOVA. Additionally, analyses of emulsifying ability (EAI) and stability (ESI) of NOVA and POVA-2 displayed that both EAI and ESI of them were significantly improved, with POVA-2 exhibiting superior EAI compared to NOVA. Therefore, this study demonstrated CP is a promising technique to promote the generation of protein amyloid fibrils in a more efficient manner.
作者机构：
[Linlin Yang] College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China;[Minglang Cai; Lei Zhong; Yi Hu; Junzhi Zhang] College of Fisheries, Hunan Agricultural University, Changsha 410128, China;Laboratory of Animal Nutritional Physiology and Metabolic Process, Key Laboratory of Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China;[Yonghong Xie] Key Laboratory of Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China;[Shouqi Xie] State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, The Chinese Academy of Sciences, Wuhan 430072, China
通讯机构：
[Yi Hu] C;College of Fisheries, Hunan Agricultural University, Changsha 410128, China
摘要：
The impacts of substituting dietary soybean meal (SBM) with yellow mealworm meal (YMM) were investigated during a 56-day feeding trial on growth, antioxidant capacity, immunity, intestinal morphology, and intestinal microbiota of grass carp. A total of 750 grass carp were divided into 5 groups (3 replications per group and 50 fish per replication) with different levels of YMM: SBM (control group), H25, H50, H75, and H100, for 8 weeks. The results showed that dietary YMM significantly increased final body weight (FW), weight gain (WG), and protein efficiency ratio (PER) in H25 group ( P < 0.05), however, complete substitution showed the opposite trend ( P < 0.05 for FW and WG). The liver antioxidant capacity was improved, manifested by enhanced superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activities, glutathione (GSH) content and up-regulated antioxidant-related genes mediated by the Keap1-Nrf2 signaling pathway in the H25 group ( P < 0.05). However, in the H100 group oxidative stress occurred in parallel with impairment of hepatic function. Intestinal inflammation was aggravated in the H100 group as evidenced by the up-regulated pro-inflammatory gene expression mediated by the NF-κB pathway ( P < 0.05). Additionally, the activity of intestinal digestive enzymes for the grass carp was significantly reduced and accompanied with intestinal mucosal barrier dysfunction in the H100 group ( P < 0.05). In summary, replacement of SBM with 25% YMM showed positive influences on growth, antioxidant capacity, immunity, and intestinal health. Conversely, complete replacement of SBM with YMM triggered oxidative stress, caused liver function disorder, and impaired intestinal health in grass carp.
The impacts of substituting dietary soybean meal (SBM) with yellow mealworm meal (YMM) were investigated during a 56-day feeding trial on growth, antioxidant capacity, immunity, intestinal morphology, and intestinal microbiota of grass carp. A total of 750 grass carp were divided into 5 groups (3 replications per group and 50 fish per replication) with different levels of YMM: SBM (control group), H25, H50, H75, and H100, for 8 weeks. The results showed that dietary YMM significantly increased final body weight (FW), weight gain (WG), and protein efficiency ratio (PER) in H25 group ( P < 0.05), however, complete substitution showed the opposite trend ( P < 0.05 for FW and WG). The liver antioxidant capacity was improved, manifested by enhanced superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activities, glutathione (GSH) content and up-regulated antioxidant-related genes mediated by the Keap1-Nrf2 signaling pathway in the H25 group ( P < 0.05). However, in the H100 group oxidative stress occurred in parallel with impairment of hepatic function. Intestinal inflammation was aggravated in the H100 group as evidenced by the up-regulated pro-inflammatory gene expression mediated by the NF-κB pathway ( P < 0.05). Additionally, the activity of intestinal digestive enzymes for the grass carp was significantly reduced and accompanied with intestinal mucosal barrier dysfunction in the H100 group ( P < 0.05). In summary, replacement of SBM with 25% YMM showed positive influences on growth, antioxidant capacity, immunity, and intestinal health. Conversely, complete replacement of SBM with YMM triggered oxidative stress, caused liver function disorder, and impaired intestinal health in grass carp.
通讯机构：
[Tan, B ] H;[Yin, YL ] C;[Yin, YL] Y;Hunan Agr Univ, Coll Anim Sci & Technol, Hunan Prov Key Lab Prod Qual Regulat Livestock & P, Changsha 410128, Hunan, Peoples R China.;Yuelushan Lab, Changsha 410128, Hunan, Peoples R China.
摘要：
BackgroundGut microbiota has been extensively demonstrated to modulate host lipid metabolism. Higher intramuscular fat (IMF) accumulation in Chinese indigenous breed pigs is associated with their special gut microbiota structure. However, the specific microbes and metabolic pathways responsible for lipid deposition are still poorly understood.ResultsIn the present study, a comparative analysis of the gut microbiota and metabolome in obese Ningxiang (NX) pigs and lean Duroc x Landrace x Yorkshire (DLY) pigs was conducted. The results revealed a higher abundance of gut lactobacilli and a correlation of branched-chain amino acid (BCAA) metabolism pathway in NX pigs. We proceeded to verify the roles of various lactobacilli strains originating from NX pigs in BCAA metabolism and lipids deposition in SD rats. We demonstrated that L. reuteri is a fundamental species responsible for modulating lipid deposition in NX pigs and that increased circulating levels of BCAA are positively linked to greater lipid deposition. Additionally, it has been verified that L. reuteri originating from NX pigs has the ability to improve BCAA synthesis in the gut and enhance IMF content in lean DLY pigs. The expression of genes related to lipid synthesis was also significantly upregulated.ConclusionsTaken together, our results imply that NX pig-derived L. reuteri regulates BCAA metabolism and plays a potential role in improving the meat quality of lean pig breeds through modulation of host intramuscular lipid deposition. The results provide a new strategy for improving the meat quality of commercial pigs by influencing host metabolism through supplementing dietary additives.F932jm79FQCSwcUD6yfj_aVideo AbstractConclusionsTaken together, our results imply that NX pig-derived L. reuteri regulates BCAA metabolism and plays a potential role in improving the meat quality of lean pig breeds through modulation of host intramuscular lipid deposition. The results provide a new strategy for improving the meat quality of commercial pigs by influencing host metabolism through supplementing dietary additives.F932jm79FQCSwcUD6yfj_aVideo Abstract
作者机构：
Authors to whom correspondence should be addressed.;[Yang, Xizi] Hunan Collaborative Innovation Center for Utilization of Botanical Functional Ingredients, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China;National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production, Institute of Subtropical Agriculture, The Chinese Academy of Sciences, Changsha 410125, China;[Wu, Shusong; He, Jianhua] Authors to whom correspondence should be addressed.<&wdkj&>Hunan Collaborative Innovation Center for Utilization of Botanical Functional Ingredients, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China;[Hu, Ruizhi] National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production, Institute of Subtropical Agriculture, The Chinese Academy of Sciences, Changsha 410125, China<&wdkj&>Hunan Collaborative Innovation Center for Utilization of Botanical Functional Ingredients, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China
通讯机构：
[Shusong Wu; Jianhua He] A;Authors to whom correspondence should be addressed.<&wdkj&>Hunan Collaborative Innovation Center for Utilization of Botanical Functional Ingredients, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China
摘要：
This review systematically examines the influence of oxidative stress on the reproductive function of male livestock, with a particular focus on the modulation of autophagy. Spermatogenesis, a highly precise biological process, is vulnerable to a range of internal and external factors, among which oxidative stress notably disrupts autophagic processes within the testes. This disruption results in diminished sperm quality, impaired testosterone synthesis, and compromised integrity of the blood-testis barrier. Furthermore, this review elucidates the molecular mechanisms by which oxidative stress-induced autophagy dysfunction impairs spermatogenesis and mitochondrial function, consequently reducing sperm motility. These findings aim to provide a theoretical foundation and serve as a reference for improving reproductive performance and sperm quality in livestock.
期刊：
Annals of Agricultural Sciences,2025年:100377 ISSN：0570-1783
通讯作者：
Qian Jiang<&wdkj&>Lijuan Chen
作者机构：
[Lijuan Chen] The Biological Feedstuff Laboratory, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, Anhui, China;Anhui Provincial Key Laboratory of Livestock and Poultry Product Safety, Institute of Animal Husbandry and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei 230011, China;[Jiashun Chen; Qian Jiang; Yulong Yin] Animal Nutritional Genome and Germplasm Innovation Research Center, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, Hunan, China;Guiding County Animal Husbandry and Aquatic Affairs Center, Chenzhou 423500, Hunan, China;[Chen Zhang] The Biological Feedstuff Laboratory, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, Anhui, China<&wdkj&>Anhui Provincial Key Laboratory of Livestock and Poultry Product Safety, Institute of Animal Husbandry and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei 230011, China<&wdkj&>Animal Nutritional Genome and Germplasm Innovation Research Center, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, Hunan, China
通讯机构：
[Qian Jiang] A;[Lijuan Chen] T;Animal Nutritional Genome and Germplasm Innovation Research Center, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, Hunan, China<&wdkj&>The Biological Feedstuff Laboratory, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, Anhui, China
关键词：
Bound gossypol;Liver injury;Sperm;Reproductive function
摘要：
Gossypol is an anti-nutritional factor in cottonseed products, and its toxicity is reduced mainly by transforming free gossypol (FG) to bound gossypol (BG). However, the safety of BG chronic consumption in relation to liver and reproduction has not been fully examined. This study aimed to assess the effects of chronic BG exposure on liver and male reproductive function. BG with a purity of 97.38 % was prepared from whole egg solution and acetate gossypol, and its digestive stability was investigated by in vitro and in vivo digestion experiments. In vitro , digestion experiment of BG in the simulated gastric and intestinal fluid was performed. In vivo , thirty 8-week-old male C57BL/6 mice were randomly assigned into 3 groups, and the effects of BG on the liver and reproductive function of mice were studied by administering 24.8 mg/(kg·BW) BG (contains 0.67 mg FG) (marked as BG group), through gavage for 90 days, and administrated with 0.67 mg/(kg·BW) FG as a control group (C group), administrated with vehicle as a negative control group. Results show that BG dissociated into FG in simulated gastrointestinal environments. Long-term BG administration hindered the growth performance of the male mice, during which higher FG was detected in the feces of BG-administrated mice. This BG administration also increased serum alanine aminotransferase and aspartate aminotransferase levels, as well as the balloon degeneration and binucleate cells in the liver. Sperm concentration, sperm motility, epididymis coefficient, and the lactate dehydrogenase-X activity in the testis were inhibited by BG administration. Taken together, we reported that long-term administration with BG at a low dosage of 24.8 mg/(kg·BW) damages the growth performance, liver, and reproductive function, which may guide the modest use of cottonseed in foods and feeds.
Gossypol is an anti-nutritional factor in cottonseed products, and its toxicity is reduced mainly by transforming free gossypol (FG) to bound gossypol (BG). However, the safety of BG chronic consumption in relation to liver and reproduction has not been fully examined. This study aimed to assess the effects of chronic BG exposure on liver and male reproductive function. BG with a purity of 97.38 % was prepared from whole egg solution and acetate gossypol, and its digestive stability was investigated by in vitro and in vivo digestion experiments. In vitro , digestion experiment of BG in the simulated gastric and intestinal fluid was performed. In vivo , thirty 8-week-old male C57BL/6 mice were randomly assigned into 3 groups, and the effects of BG on the liver and reproductive function of mice were studied by administering 24.8 mg/(kg·BW) BG (contains 0.67 mg FG) (marked as BG group), through gavage for 90 days, and administrated with 0.67 mg/(kg·BW) FG as a control group (C group), administrated with vehicle as a negative control group. Results show that BG dissociated into FG in simulated gastrointestinal environments. Long-term BG administration hindered the growth performance of the male mice, during which higher FG was detected in the feces of BG-administrated mice. This BG administration also increased serum alanine aminotransferase and aspartate aminotransferase levels, as well as the balloon degeneration and binucleate cells in the liver. Sperm concentration, sperm motility, epididymis coefficient, and the lactate dehydrogenase-X activity in the testis were inhibited by BG administration. Taken together, we reported that long-term administration with BG at a low dosage of 24.8 mg/(kg·BW) damages the growth performance, liver, and reproductive function, which may guide the modest use of cottonseed in foods and feeds.
摘要：
Eimeria tenella (E. tenella) infection is a major cause of coccidiosis in chickens, leading to significant economic losses in the poultry industry due to its impact on the cecum. This study presents a comprehensive single-cell atlas of the chicken cecal epithelium by generating 7,394 cells using 10X Genomics single-cell RNA sequencing (scRNA-seq). We identified 13 distinct cell types, including key immune and epithelial populations, and characterized their gene expression profiles and cell-cell communication networks. Integration of this single-cell data with bulk RNA-seq data from E. tenella-infected chickens revealed significant alterations in cell type composition and state, particularly a marked decrease in APOB(+) enterocytes and an increase in cycling T cells during infection. Trajectory analysis of APOB(+) enterocytes uncovered shifts toward cellular states associated with cell death and a reduction in those linked to mitochondrial and cytoplasmic protection when infected with E. tenella. These findings highlight the substantial impact of E. tenella on epithelial integrity and immune responses, emphasizing the parasite's role in disrupting nutrient absorption and energy metabolism. Our single-cell atlas serves as a critical resource for understanding the cellular architecture of the chicken cecum and provides a valuable framework for future investigations into cecal diseases and metabolic functions, with potential applications in enhancing poultry health and productivity.
摘要：
This study aimed to investigate the effects of dietary isatidis root polysaccharide (IRP) on diarrhea, immunity, and intestinal health in weanling piglets. Forty healthy piglets were randomly assigned to five groups receiving varying dosages of IRP. The findings indicated that different concentrations of IRP significantly reduced diarrhea scores (p < 0.01). Notably, the serum levels of immunoglobulin A and immunoglobulin G increased linearly and quadratically (p < 0.01), while immunoglobulin M also showed a linear increase (p < 0.05) in IRP-fed piglets. The secretory immunoglobulin A levels in ileal contents were significantly higher compared to control piglets (p < 0.01). Key intestinal health parameters, including villus height, villus height-to-crypt depth ratio, and goblet cell numbers, showed linear and quadratic increases in both the jejunum and ileum (p < 0.05), while crypt depth decreased significantly (p < 0.01). Additionally, the expression of IL-10, ZO-1, occludin, and mucin2 was upregulated linearly and quadratically in IRP-fed piglets (p < 0.05). In cultured IPEC-J2 cells, ZO-1 and occludin expression levels significantly increased upon exposure to 400 μg/mL IRP (p < 0.01). Furthermore, the relative abundances of Escherichia coli, Ralstonia pickettii, and Desulfovibrio fairfieldensis decreased linearly with increasing dietary IRP concentration. In conclusion, IRP shows promise as an effective dietary supplement for mitigating diarrhea and enhancing intestinal health in early weaned piglets.
作者机构：
[Yang, Hong; Fu, Jiaojiao; Zhang, Mengyuan; Cao, Xinyi; Xiao, Yu] Fisheries College, Hunan Agricultural University, Changsha 410128, China;Hunan Engineering Technology Research Center of Featured Aquatic Resources Utilization, Hunan Agricultural University, Changsha 410128, China;[Wang, Hongquan; Xiao, Tiaoyi; Lv, Zhao] Fisheries College, Hunan Agricultural University, Changsha 410128, China<&wdkj&>Hunan Engineering Technology Research Center of Featured Aquatic Resources Utilization, Hunan Agricultural University, Changsha 410128, China
通讯机构：
[Zhao Lv] F;Fisheries College, Hunan Agricultural University, Changsha 410128, China<&wdkj&>Hunan Engineering Technology Research Center of Featured Aquatic Resources Utilization, Hunan Agricultural University, Changsha 410128, China
摘要：
IL-10, an anti-inflammatory cytokine, plays crucial roles in the context of infectious inflammatory diseases, whose gene variations and expression are tightly linked to disease resistance in animals. However, there is limited research on the genetic variations governing IL-10 expression and the underlying effects in fish. In this study, we identified a GAGTT insertion/deletion (Ins/Del) variation within the IL-10 gene promoter and analyzed the effects of this variation on IL-10 expression, inflammatory response and its association with disease resistance in grass carp. With dual luciferase reporter assays, we demonstrated that the GAGTT Ins/Del variation significantly altered the IL-10 promoter activity in grass carp. DNA pull-down combined with mass spectrometry revealed that the spectrum of transcription factors that bind to IL-10 promoter variants caused by GAGTT Ins/Del variation was quite different. Moreover, three representative transcription factors, JunD/AP-1, p50/NF-κB, and HMGB1, differentially transactivated grass carp IL-10 promoter variants, as confirmed by western blotting and dual luciferase reporter assays. Using the GAGTT/− genotype of the heterozygous grass carp parents, we obtained full-sib family offspring of the GAGTT/GAGTT, GAGTT/−, and −/− genotypes. During grass carp reovirus (GCRV) infection, these three IL-10 genotypes of grass carp offspring displayed different survival rates. Importantly, grass carp with the GAGTT/− genotype had greater survival than those with the other two genotypes after GCRV infection. Furthermore, the results indicated that the improved intestinal inflammation at 1 d post-GCRV infection and enhanced immunocompetence in the spleen at 5 d post-GCRV infection may contribute to the disease resistance of grass carp with the GAGTT/− genotype. Overall, this study offers new insight into the inflammatory and immune regulatory mechanisms specific to IL-10 from a genetic perspective in fish and may provide a valuable molecular marker for breeding disease-resistant grass carp.
IL-10, an anti-inflammatory cytokine, plays crucial roles in the context of infectious inflammatory diseases, whose gene variations and expression are tightly linked to disease resistance in animals. However, there is limited research on the genetic variations governing IL-10 expression and the underlying effects in fish. In this study, we identified a GAGTT insertion/deletion (Ins/Del) variation within the IL-10 gene promoter and analyzed the effects of this variation on IL-10 expression, inflammatory response and its association with disease resistance in grass carp. With dual luciferase reporter assays, we demonstrated that the GAGTT Ins/Del variation significantly altered the IL-10 promoter activity in grass carp. DNA pull-down combined with mass spectrometry revealed that the spectrum of transcription factors that bind to IL-10 promoter variants caused by GAGTT Ins/Del variation was quite different. Moreover, three representative transcription factors, JunD/AP-1, p50/NF-κB, and HMGB1, differentially transactivated grass carp IL-10 promoter variants, as confirmed by western blotting and dual luciferase reporter assays. Using the GAGTT/− genotype of the heterozygous grass carp parents, we obtained full-sib family offspring of the GAGTT/GAGTT, GAGTT/−, and −/− genotypes. During grass carp reovirus (GCRV) infection, these three IL-10 genotypes of grass carp offspring displayed different survival rates. Importantly, grass carp with the GAGTT/− genotype had greater survival than those with the other two genotypes after GCRV infection. Furthermore, the results indicated that the improved intestinal inflammation at 1 d post-GCRV infection and enhanced immunocompetence in the spleen at 5 d post-GCRV infection may contribute to the disease resistance of grass carp with the GAGTT/− genotype. Overall, this study offers new insight into the inflammatory and immune regulatory mechanisms specific to IL-10 from a genetic perspective in fish and may provide a valuable molecular marker for breeding disease-resistant grass carp.
期刊：
JOURNAL OF ANIMAL BREEDING AND GENETICS,2025年142(2):223-236 ISSN：0931-2668
通讯作者：
Zhang, Z
作者机构：
[Wei, Chen; Gao, Yahui; Li, Jiaqi; Zhang, Zhe; Cai, Xiaodian; Zhang, Wenjing] South China Agr Univ, Coll Anim Sci, Natl Engn Res Ctr Breeding Swine Ind, Guangdong Prov Key Lab Agroanim Genom & Mol Breed, Guangzhou, Peoples R China.;[Gao, Ning] Hunan Agr Univ, Coll Anim Sci & Technol, Changsha, Peoples R China.;[Wu, Xibo; Si, Jinglei] Guangxi State Farmd Yongxin Anim Husb Grp Co Ltd, Nanning, Peoples R China.;[Yin, Tong] Justus Liebig Univ, Inst Anim Breeding & Genet, Giessen, Germany.;[Zhang, Z] South China Agr Univ, Dept Anim Breeding & Genet, Guangzhou 510642, Peoples R China.
通讯机构：
[Zhang, Z ] S;South China Agr Univ, Dept Anim Breeding & Genet, Guangzhou 510642, Peoples R China.
关键词：
combined population;genomic prediction;heritability enrichment;meta-analysis of genome-wide association studies
摘要：
The strategy of combining reference populations has been widely recognized as an effective way to enhance the accuracy of genomic prediction (GP). This study investigated the efficiency of genomic prediction using prior information and combined reference population. In total, prior information considering trait-associated single nucleotide polymorphisms (SNPs) obtained from meta-analysis of genome-wide association studies (GWAS meta-analysis) was incorporated into three models to assess the performance of GP using combined reference populations. Two different Yorkshire populations with imputed whole genome sequence (WGS) data (9,741,620 SNPs), named as P1 (1259 individuals) and P2 (1018 individuals), were used to predict genomic estimated breeding values for three live carcass traits, including backfat thickness, loin muscle area, and loin muscle depth. A 10 × 5 fold cross-validation was used to evaluate the prediction accuracy of 203 randomly selected candidate pigs from the P2 population and the reference population consisted of the remaining pigs from P2 and the stepwise added pigs from P1. By integrating SNPs with different p-value thresholds from GWAS meta-analysis downloaded from PigGTEx Project, the prediction accuracy of GBLUP, genomic feature BLUP (GFBLUP) and GBLUP given genetic architecture (BLUP|GA) were compared. Moreover, we explored effects of reference population size and heritability enrichment of genomic features on the prediction accuracy improvement of GFBLUP and BLUP|GA relative to GBLUP. The prediction accuracy of GBLUP using all WGS markers showed average improvement of 4.380% using the P1 + P2 reference population compared with the P2 reference population. Using the combined reference population, GFBLUP and BLUP|GA yielded 6.179% and 5.525% higher accuracies than GBLUP using all SNPs based on the single reference population, respectively. Positive regression coefficients were estimated in relation to the improvement in prediction accuracy (between GFBLUP/BLUP|GA and GBLUP) and the size of the reference as well as the heritability enrichment of genomic features. Compared to the classic GBLUP model, GFBLUP and BLUP|GA models integrating GWAS meta-analysis information increase the prediction accuracy and using combined populations with enlarged reference population size further enhances prediction accuracy of the two approaches. The heritability enrichment of genomic features can be used as an indicator to reflect weather prior information is accurately presented.
摘要：
Machine learning (ML) methods have rapidly developed in various theoretical and practical research areas, including predicting genomic breeding values for large livestock animals. However, few studies have investigated the application of ML in broiler breeding. In this study, seven different ML methods—support vector regression (SVR), random forest (RF), gradient boosting decision tree (GBDT), extreme gradient boosting (XGBoost), light gradient boosting machine (LightGBM), kernel ridge regression (KRR) and multilayer perceptron (MLP) were employed to predict the genomic breeding values of laying traits, growth and carcass traits in a yellow-feathered broiler breeding population. The results indicated that classic methods, such as GBLUP and Bayesian, achieved superior prediction accuracy compared to ML methods in five of the eight traits. For half-eviscerated weight (HEW), ML methods showed an average improvement of 54.4% over GBLUP and Bayesian methods. Among the ML methods, SVR, RF, GBDT, and XGBoost exhibited improvements exceeding 60%, with respective values of 61.3%, 61.0%, 60.4%, and 60.7%; while MLP improved by 54.4% and LightGBM by 53.7%, KRR had the lowest improvement at 29.4%. For eviscerated weight (EW), ML methods still outperformed GBLUP and Bayesian methods. MLP gained the largest improvement at 19.0%, while SVR, RF, GBDT, XGBoost, LightGBM, and KRR improved by 15.0%, 16.5%, 9.5%, 7.0%, 1.6%, and 15.9%, respectively. Compared to default hyperparameters, the average improvement of ML methods with tuned hyperparameters was 34.0%, 32.9%, 27.0%, 19.3%, 26.8%, 13.2%, 18.9%, and 46.3%, respectively. The prediction accuracy of above algorithms could be optimized using genome-wide association study (GWAS) to select subsets of significant SNPs. This work provides valuable insights into genomic prediction, aiding genetic breeding in broilers.
Machine learning (ML) methods have rapidly developed in various theoretical and practical research areas, including predicting genomic breeding values for large livestock animals. However, few studies have investigated the application of ML in broiler breeding. In this study, seven different ML methods—support vector regression (SVR), random forest (RF), gradient boosting decision tree (GBDT), extreme gradient boosting (XGBoost), light gradient boosting machine (LightGBM), kernel ridge regression (KRR) and multilayer perceptron (MLP) were employed to predict the genomic breeding values of laying traits, growth and carcass traits in a yellow-feathered broiler breeding population. The results indicated that classic methods, such as GBLUP and Bayesian, achieved superior prediction accuracy compared to ML methods in five of the eight traits. For half-eviscerated weight (HEW), ML methods showed an average improvement of 54.4% over GBLUP and Bayesian methods. Among the ML methods, SVR, RF, GBDT, and XGBoost exhibited improvements exceeding 60%, with respective values of 61.3%, 61.0%, 60.4%, and 60.7%; while MLP improved by 54.4% and LightGBM by 53.7%, KRR had the lowest improvement at 29.4%. For eviscerated weight (EW), ML methods still outperformed GBLUP and Bayesian methods. MLP gained the largest improvement at 19.0%, while SVR, RF, GBDT, XGBoost, LightGBM, and KRR improved by 15.0%, 16.5%, 9.5%, 7.0%, 1.6%, and 15.9%, respectively. Compared to default hyperparameters, the average improvement of ML methods with tuned hyperparameters was 34.0%, 32.9%, 27.0%, 19.3%, 26.8%, 13.2%, 18.9%, and 46.3%, respectively. The prediction accuracy of above algorithms could be optimized using genome-wide association study (GWAS) to select subsets of significant SNPs. This work provides valuable insights into genomic prediction, aiding genetic breeding in broilers.
摘要：
The complement system, a critical component of innate immunity in fish, plays a pivotal role in the defense against Grass Carp Reovirus (GCRV) infection in grass carp. This study explores the structural characteristics of C1S , a crucial molecule in the classical pathway of the complement system, and its involvement in the response to GCRV infection. We found that the grass carp C1S gene comprises six domains similar to those in mammals: two CUB (Complement C1r/C1s, Uegf, Bmp1) domains, two CCP (Complement control protein) domains, one EGFCA (Calcium-binding epidermal growth factor) domain, and one Tryp_SPc (Trypsin‐like serine protease) domain, albeit without chromosomal collinearity to humans. Comparative analysis revealed that the identity and similarity of this gene with those in other species range from 30.6–89.4% and 30.7–89.7%, respectively. Phylogenetic analysis positioned C1S in close relation with R. klamathensis and D. rerio . Tissue expression profiles in both healthy and GCRV-infected grass carp indicated primary expression of C1S in the liver, with expression peaks at day 7 post-infection in the liver and spleen, and at day 5 in the kidney. Functional assays demonstrated that C1S activates the complement system via cleavage of complement component 3 (C3) into C3b, further inhibiting GCRV replication and upregulating antiviral genes IFN1 , IRF3 , and IRF7 . These findings elucidate the mechanism by which the complement system mediates resistance to GCRV infection in grass carp, offering a substantial theoretical foundation for further research.
The complement system, a critical component of innate immunity in fish, plays a pivotal role in the defense against Grass Carp Reovirus (GCRV) infection in grass carp. This study explores the structural characteristics of C1S , a crucial molecule in the classical pathway of the complement system, and its involvement in the response to GCRV infection. We found that the grass carp C1S gene comprises six domains similar to those in mammals: two CUB (Complement C1r/C1s, Uegf, Bmp1) domains, two CCP (Complement control protein) domains, one EGFCA (Calcium-binding epidermal growth factor) domain, and one Tryp_SPc (Trypsin‐like serine protease) domain, albeit without chromosomal collinearity to humans. Comparative analysis revealed that the identity and similarity of this gene with those in other species range from 30.6–89.4% and 30.7–89.7%, respectively. Phylogenetic analysis positioned C1S in close relation with R. klamathensis and D. rerio . Tissue expression profiles in both healthy and GCRV-infected grass carp indicated primary expression of C1S in the liver, with expression peaks at day 7 post-infection in the liver and spleen, and at day 5 in the kidney. Functional assays demonstrated that C1S activates the complement system via cleavage of complement component 3 (C3) into C3b, further inhibiting GCRV replication and upregulating antiviral genes IFN1 , IRF3 , and IRF7 . These findings elucidate the mechanism by which the complement system mediates resistance to GCRV infection in grass carp, offering a substantial theoretical foundation for further research.
摘要：
The inclusion of Macleaya cordata extract (MCE) and antimicrobial peptides (AMP) in animal feed has been shown to improve performance, potentially providing synergistic benefits when applied in combination. This study aims to investigate the potential synergistic effects of MCE and AMP supplementation on reproductive performance and oviduct microbial composition in Xuefeng black-bone chickens. A total of 900 chickens were randomly assigned to 5 groups with 6 replicates of 30 hens. The group A received a basal diet, while the experimental groups (groups B, C, D, and E) were supplemented with either 500 or 1,000 mg/kg of MCE and 200 or 400 mg/kg of AMP for 66 days. The results demonstrated that (1) Group B exhibited a significant increase in laying rate compared to group A ( p < 0.05), along with a significant reduction in feed conversion rate ( p < 0.05). Moreover, group E exhibited a significant increase in average daily egg weight ( p < 0.05), and all experimental groups showed a significant reduction in abnormal egg rates ( p < 0.05). (2) Groups B and C exhibited significantly thicker oviduct epithelium compared to group A ( p < 0.05). (3) At the phylum level, all dietary treatment groups showed a higher relative abundance of Firmicutes compared to group A, with the highest levels found in group B ( p < 0.05). (4) At the genus level, the relative abundance of Faecalibacterium in groups C and E was significantly higher than in group A ( p < 0.05).
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Previous studies have shown that adding 450 mg/kg of ursolic acid (UA) can improve the growth performance of broilers. However, the specific mechanism is still unclear. Therefore, the purpose of this study was to further explore whether UA promotes the growth of broilers by affecting the intestinal environment of broilers. We randomly divided 120 broilers with similar BW (46.53 ± 0.05 g) into two groups. Each group had six replicates, with 10 broilers per replicate. The broilers were fed either the corn-soybean meal−basal diet (CON group) or the corn-soybean meal−basal diet supplemented with 450 mg/kg UA (UA group). This study lasted 42 days. Adding UA increased the daily weight gain and feed conversion ratio of broilers (P < 0.05). The UA group exhibited reduced aspartate aminotransferase, total cholesterol, interleukin 6 and interleukin 1, and triacylglycerol levels, with increased interleukin 10 and high-density lipoprotein cholesterol in serum (P < 0.05). The UA supplementation improved total antioxidant capacity, total superoxide dismutase, and glutathione peroxidase activity in serum (P < 0.05), and increased these levels in the jejunum (P < 0.05). It reduced malondialdehyde concentration in the jejunum and ileum (P < 0.05), improved jejunal morphology by increasing villus height and villus-to-crypt ratio, and decreased crypt depth (P < 0.05). Gene expression of zona occludens 1 and Claudin-1 was higher, while interleukin 6 was lower in the UA group (P < 0.05). Additionally, interleukin 10 gene expression in jejunal mucosa was higher (P < 0.05). Significant differences were observed in the abundance of Bacteroides, proteobacteria, and desulfurisation bacteria (P < 0.05), with higher Barnesiella and Clostridia_UCG-014, and lower Romboutsia in the UA group (P < 0.05). Barnesiella negatively correlated with interleukin 6, interleukin 1, and triacylglycerol, but positively correlated with interleukin 10 (P < 0.05). In conclusion, adding 450 mg/kg UA to broiler feed can improve serum and jejunal antioxidant capacity, reduce jejunal and ileal inflammation, improve jejunal morphology, and regulate caecal microbiota structure composition, promoting broiler growth.
Previous studies have shown that adding 450 mg/kg of ursolic acid (UA) can improve the growth performance of broilers. However, the specific mechanism is still unclear. Therefore, the purpose of this study was to further explore whether UA promotes the growth of broilers by affecting the intestinal environment of broilers. We randomly divided 120 broilers with similar BW (46.53 ± 0.05 g) into two groups. Each group had six replicates, with 10 broilers per replicate. The broilers were fed either the corn-soybean meal−basal diet (CON group) or the corn-soybean meal−basal diet supplemented with 450 mg/kg UA (UA group). This study lasted 42 days. Adding UA increased the daily weight gain and feed conversion ratio of broilers (P < 0.05). The UA group exhibited reduced aspartate aminotransferase, total cholesterol, interleukin 6 and interleukin 1, and triacylglycerol levels, with increased interleukin 10 and high-density lipoprotein cholesterol in serum (P < 0.05). The UA supplementation improved total antioxidant capacity, total superoxide dismutase, and glutathione peroxidase activity in serum (P < 0.05), and increased these levels in the jejunum (P < 0.05). It reduced malondialdehyde concentration in the jejunum and ileum (P < 0.05), improved jejunal morphology by increasing villus height and villus-to-crypt ratio, and decreased crypt depth (P < 0.05). Gene expression of zona occludens 1 and Claudin-1 was higher, while interleukin 6 was lower in the UA group (P < 0.05). Additionally, interleukin 10 gene expression in jejunal mucosa was higher (P < 0.05). Significant differences were observed in the abundance of Bacteroides, proteobacteria, and desulfurisation bacteria (P < 0.05), with higher Barnesiella and Clostridia_UCG-014, and lower Romboutsia in the UA group (P < 0.05). Barnesiella negatively correlated with interleukin 6, interleukin 1, and triacylglycerol, but positively correlated with interleukin 10 (P < 0.05). In conclusion, adding 450 mg/kg UA to broiler feed can improve serum and jejunal antioxidant capacity, reduce jejunal and ileal inflammation, improve jejunal morphology, and regulate caecal microbiota structure composition, promoting broiler growth.
摘要：
The poultry industry relies heavily on soybean meal to feed egg-laying chickens, but its high cost and price instability necessitate the exploration of more affordable and reliable alternatives. Fish bone meal, a nutritious byproduct from fish processing, offers high-quality protein and essential minerals, making it a promising addition to poultry diets. In this study, 240 Hy-Line Brown laying hens (31 weeks old, average weight 1.90 ± 0.1 kg, egg laying 11 weeks) were randomly assigned to two dietary treatments for 12 weeks: one group received a regular diet, while the other was fed a diet containing 3% fish bone meal. Each treatment included 12 replicates with 10 hens per replicate, and all birds were maintained under identical environmental conditions using a completely randomized experimental design. The results showed that hens fed the fish bone meal diet produced more eggs (p < 0.05) and had heavier yolks (p < 0.05) compared to the control group. Furthermore, intestinal health indicators, including villus height and crypt depth, were significantly improved (p < 0.05), along with enhanced immune markers (p < 0.05) in the fish bone meal group. However, no significant differences were observed in other egg quality parameters or feed conversion efficiency between the two groups (p > 0.05). These findings demonstrate that fish bone meal can improve egg production, yolk quality, intestinal health, and immune function in laying hens.
关键词：
carcass traits;fatty-type pig;gut microbiota;low protein diet;meat quality
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INTRODUCTION: Soybean meal is an excellent protein source and is widely used in pig feed. However, the Americas account for more than 80% of global soybean production, so European and Asia swine production largely depends on soybean imports. The use of safe and functional unconventional feed sources can effectively alleviate worldwide protein shortage problems. METHODS: Here, we formulated a low-protein soybean-free diet (LPNS) for growing and fattening pigs using rice, potatoes, tea, and other unconventional feed sources. Thirty-six healthy Daweizi pigs (average body weight 23.60 ± 1.34 kg) were raised under the same conditions and randomly assigned to two dietary treatments: (1) Con group, corn-soybean base meal; (2) LPNS group. When the average weight of pigs in the group reached 85 kg, two pigs per pen were randomly selected and euthanized for collection of the colonic digesta and carcass traits and for meat quality determination. RESULTS: Compared with the corn-soybean based diet, the LPNS diet decreased the average daily gain (ADG) and feed conversion ratio (FCR) of Daweizi pigs but had a lower cost per kilogram of gain. In addition, the LPNS diet significantly increased leanness and decreased the fat-skin rate and bone rate of Daweizi pigs. The cooking loss of meat decreased, and unsaturated fatty acids such as C22:6 and n-3 PUFA significantly increased in the LPNS group. Moreover, the purine content in the meat substantially decreased with the LPNS diet. The 16S rDNA analysis revealed that the LPNS diet greatly modified the composition of the colonic microbiota community, with a decrease in the Firmicutes/Bacteroidetes ratio and an increase in the abundance of Lactobacillus spp. DISCUSSION: The use of functional herbs along with a low-protein diet helped to regulate fat and purine metabolism in fatty-type pigs. The LPNS diet formulated with unconventional-feed sources not only helps reduce the feed cost in swine production but also improves the carcass traits and meat quality of pigs, which is more suitable for small-scale pig farming.
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The favorable impacts of Bacillus coagulans or Yeast culture on broiler production performance and immune function have been acknowledged. However, the effects of the combined of them (BcYc) on the broilers remained unknown. Thus, the current research aimed to assess the effects of BcYc (at dosages of 0, 200, 300, and 400mg/kg) on growth performance, plasma biochemical indices, antioxidant capacity, and intestinal morphology and microbial composition in broilers. The results revealed that ①broilers receiving 300 and 400mg/kg of BcYc showed significantly higher body weights than the control group at 4d and 21d (P < 0.05). Additionally, when contrasted with the control group, the feed conversion ratio was significantly decreased in 300mg/kg group during 4 to 21d and in 400mg/kg group during 4 to 42d (P < 0.05). ②At 21d, the broilers supplemented with 300 and 400mg/kg of BcYc demonstrated significantly elevated levels of albumin (ALB), glutathione peroxidase activity (GSH-Px), total antioxidant capacity, and catalase activity than the group that not supplemented with BcYc (P < 0.05). At 42d, 400mg/kg group showed significantly higher ALB and total protein, and lower glutamic-pyruvic transaminase activity and malondialdehyde content in contrast to the control group (P < 0.05). Furthermore, at 42d, GSH-Px activity in all experimental groups showed significantly higher compared to the control group (P < 0.05). ③Nevertheless, no significant variation was observed neither in jejunum or duodenum morphology among all groups (P > 0.05). ④Adding 400mg/kg of BcYc led to a significantly higher Sobs index and a lower Simpson index (P < 0.05), and an extremely significantly raised in Shannon index (P < 0.01), while adding 300mg/kg of BcYc led to a significantly enhanced in Shannon index (P < 0.05). Additionally, supplementary BcYc significantly elevated the abundance of Firmicutes in phylum level, and Clostridia, Ruminococcus, Rikenella, Butyricicoccus and Eubacterium in genus level (P < 0.05). To sum up, supplementing BcYc at dosages of 400mg/kg to yellow-feathered broilers diet can help improve the growth performance, regulate plasma biochemical indicators, increase antioxidant capacity, and improve gut community composition.
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Maternal nutrition is one of the main factors regulating the growth and immunity of piglets. This study aimed to investigate the effects of maternal or maternal-offspring supplementation of antibiotics, probiotics, and synbiotics on the immunity of offspring (21, 65, and 125 day-old) in Bama mini-pigs. The results showed that adding antibiotics to maternal diets increased the plasma IFN-gamma level of offspring piglets at 21 day-old. Compared with maternal supplementation, maternal-offspring supplementation of antibiotics decreased the IL-10 level in the spleen, probiotics decreased IL-2, IL-10, and TNF-alpha levels in the ileum, and synbiotics decreased IL-10 and IFN-gamma levels in the ileum of offspring piglets. Moreover, maternal-offspring antibiotics supplementation increased the IL-1 beta level in the ileum, while probiotics supplementation increased the IL-1 beta level in the spleen of offspring piglets. Maternal antibiotics supplementation increased the TNF-alpha level in the ileum at 95 day-old compared with maternal probiotics and synbiotics supplementation. Maternal-offspring antibiotics supplementation increased the IL-1 beta level in the ileum compared with the probiotics supplementation, while synbiotics supplementation increased the IL-6 level in the ileum than the probiotics and antibiotics supplementation at 95 day-old. Moreover, maternal-offspring probiotics supplementation increased the IL-1 beta level in the spleen of offspring pigs, which was higher than the maternal probiotics supplementation. These findings suggest that the immune function of the offspring piglets varied depending on the specific approach used for probiotics and synbiotics supplementation.
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Superoxide dismutase (SOD) is a vital antioxidant enzyme that exerts antioxidative and anti-inflammatory effects on the host. In this study, a novel thermostable SOD of Alicyclobacillus sp. (AliSOD) from a hot spring was overexpressed in Escherichia coli, and enzymatic properties were identified. Mn2+ plays a decisive role in enzyme activity, indicating that AliSOD is MnSOD. Specifically, AliSOD was determined to be dimeric with a subunit molecular mass of 23.0 kDa, and the specific activity was confirmed to be as high as 24990.8 U<middle dot>mg(-1). AliSOD demonstrated exceptional thermal stability, broad pH stability, and resistance to urea, exhibiting minimal loss of activity at 70 degrees C and remarkable tolerance in an alkaline environment. Moreover, AliSOD significantly alleviated oxidative stress in diquat-injured cells (P < 0.01). It also increased intracellular SOD expression and activated the Nrf2 protein downstream of the Keap1-Nrf2 signaling pathway (P < 0.05). Overall, AliSOD exhibits excellent thermostability and specific activity, indicating potential applications in the pharmaceutical, food, and animal feed industries.
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Stilbenoids are a class of naturally occurring phenolic compounds found in various plant species, characterized by a stilbene backbone with diverse substituents that confer a range of biological activities. These compounds exhibit antioxidant, anti-inflammatory, and antimicrobial properties, making them promising candidates for improving intestinal health. The intestinal tract plays a critical role in nutrient digestion, absorption, and immune defense, and maintaining its integrity is vital for animal growth. Stilbenoids contribute to gut health by enhancing intestinal morphology, supporting mucosal immune responses, regulating gut microbiota composition, modulating metabolic pathways, and maintaining mitochondrial health. This review provides a comprehensive analysis of key stilbenoids, including resveratrol, pterostilbene, piceatannol, and oxyresveratrol, focusing on their biological effects and regulatory mechanisms. By highlighting their roles in mitigating intestinal inflammation and promoting gut function, this review provides a basis for the practical application of stilbenoids in animal health and husbandry.
