Keayla M Harr,Madelyn A Scott,Eduardo Solano Pina Dos Santos,Nara R B Cônsolo,Logan Johnson,Gretchen G Mafi,Morgan M Pfeiffer,Ranjith Ramanathan
Dark-cutting beef continues to remain one of the challenges for the global beef industry. The objective of this study was to determine the impact of varying visual degrees of dark-cutting condition on the lipidome profiles of beef longissimus lumborum. Beef carcasses (n = 6/treatment; 24 total loins) were identified at the time of grading based on the visual severity of dark-cutting levels, and beef strip loins (longissimus lumborum) were collected from these carcasses following fabrication. Treatments included a normal, bright cherry red control (pH = 5.54), shady dark-cutting (half dark; pH = 5.96), moderate dark-cutting (two-thirds dark; pH = 6.38), and moderately severe dark-cutting (full dark; pH = 6.55). Approximately 48-60 h postmortem, a single steak was sliced off the anterior end of each loin for bloom color analysis and nontargeted lipidomic analysis using a liquid chromatography-mass-spectrometry approach. A total of 379 lipids, representing different classes of lipids, were identified across the four treatments. Pairwise comparisons revealed 21, 22, and 23 lipid species that differed (P < 0.05) in shady, moderate, and moderately severe dark-cutting beef, respectively, compared with normal beef. Only one lipid species (acylcarnitine 22:2) differed between moderately severe and moderate groups. Acylcarnitine species of varying carbons and saturations were the most common of the shared species in dark-cutting samples. Medium- and long-chain acylcarnitine levels were significantly different (P < 0.05) between dark-cutting treatments and normal pH. The importance projection analysis indicated that acylcarnitine 20:2, 18:1, and 16:0 were the top three lipid species contributing to differences between dark-cutting severities and normal steaks. The relative proportion of lipids involved in energy metabolism was greater (P < 0.05) in moderate and moderately severe than in normal bright red steaks. Greater acylcarnitine levels in postmortem muscle suggest mobilization of fatty acids for energy homeostasis in dark-cutting beef and altered metabolism.
{"title":"A preliminary nontargeted lipidomics analysis reveals greater acylcarnitine in dark-cutting beef longissimus lumborum across visual severity levels.","authors":"Keayla M Harr,Madelyn A Scott,Eduardo Solano Pina Dos Santos,Nara R B Cônsolo,Logan Johnson,Gretchen G Mafi,Morgan M Pfeiffer,Ranjith Ramanathan","doi":"10.1093/jas/skaf460","DOIUrl":"https://doi.org/10.1093/jas/skaf460","url":null,"abstract":"Dark-cutting beef continues to remain one of the challenges for the global beef industry. The objective of this study was to determine the impact of varying visual degrees of dark-cutting condition on the lipidome profiles of beef longissimus lumborum. Beef carcasses (n = 6/treatment; 24 total loins) were identified at the time of grading based on the visual severity of dark-cutting levels, and beef strip loins (longissimus lumborum) were collected from these carcasses following fabrication. Treatments included a normal, bright cherry red control (pH = 5.54), shady dark-cutting (half dark; pH = 5.96), moderate dark-cutting (two-thirds dark; pH = 6.38), and moderately severe dark-cutting (full dark; pH = 6.55). Approximately 48-60 h postmortem, a single steak was sliced off the anterior end of each loin for bloom color analysis and nontargeted lipidomic analysis using a liquid chromatography-mass-spectrometry approach. A total of 379 lipids, representing different classes of lipids, were identified across the four treatments. Pairwise comparisons revealed 21, 22, and 23 lipid species that differed (P < 0.05) in shady, moderate, and moderately severe dark-cutting beef, respectively, compared with normal beef. Only one lipid species (acylcarnitine 22:2) differed between moderately severe and moderate groups. Acylcarnitine species of varying carbons and saturations were the most common of the shared species in dark-cutting samples. Medium- and long-chain acylcarnitine levels were significantly different (P < 0.05) between dark-cutting treatments and normal pH. The importance projection analysis indicated that acylcarnitine 20:2, 18:1, and 16:0 were the top three lipid species contributing to differences between dark-cutting severities and normal steaks. The relative proportion of lipids involved in energy metabolism was greater (P < 0.05) in moderate and moderately severe than in normal bright red steaks. Greater acylcarnitine levels in postmortem muscle suggest mobilization of fatty acids for energy homeostasis in dark-cutting beef and altered metabolism.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"31 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145907715","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Circular RNAs (circRNAs) are key gene regulators that are involved in many fundamental biological processes. Skeletal muscle cannot develop normally without the involvement of circRNA. While circRNA plays a pivotal role in skeletal muscle development, its functions and mechanisms within the muscle development of local chicken breeds remain to be thoroughly elucidated. This study, using the Jingyuan chicken as a model, investigated the regulatory role of circACLY in skeletal muscle development and inosine monophosphate (IMP) synthesis. Experiments verified that circACLY is a circRNA that is mainly localized to the cytoplasm. Bioinformatics analysis and experimental validation demonstrated that circACLY is a target of gga-miR-6660-3p, which in turn regulates the expression of Ectonucleotide Triphosphate Diphosphohydrolase 7 (ENTPD7). Functional assays demonstrated that gga-miR-6660-3p inhibits myoblast proliferation and differentiation capacity while suppressing the de novo IMP synthesis pathway. However, circACLY reverses the inhibitory effects of gga-miR-6660-3p on these processes. This study systematically elucidates the regulatory mechanism of the circACLY/gga-miR-6660-3p/ENTPD7 axis in the development of quail myoblasts and IMP synthesis, providing a theoretical basis for deepening the understanding of the molecular basis of skeletal muscle development in local chicken breeds and the genetic improvement of meat quality traits.
{"title":"CircACLY regulates proliferation, differentiation and apoptosis of Jingyuan chicken myoblasts by sponging gga-miR-6660-3P.","authors":"Ruimin Ma,Ying Zhou,Weizhen Wang,Ling Zhu,Tong Zhang,Jinli Tian,Lijuan Yang,Hua Wang,Lin Xue,Siyu Chen,Xiaohua Tian,Xiaoyun Ji,Zhenyun Cai,Yaling Gu,Juan Zhang","doi":"10.1093/jas/skaf464","DOIUrl":"https://doi.org/10.1093/jas/skaf464","url":null,"abstract":"Circular RNAs (circRNAs) are key gene regulators that are involved in many fundamental biological processes. Skeletal muscle cannot develop normally without the involvement of circRNA. While circRNA plays a pivotal role in skeletal muscle development, its functions and mechanisms within the muscle development of local chicken breeds remain to be thoroughly elucidated. This study, using the Jingyuan chicken as a model, investigated the regulatory role of circACLY in skeletal muscle development and inosine monophosphate (IMP) synthesis. Experiments verified that circACLY is a circRNA that is mainly localized to the cytoplasm. Bioinformatics analysis and experimental validation demonstrated that circACLY is a target of gga-miR-6660-3p, which in turn regulates the expression of Ectonucleotide Triphosphate Diphosphohydrolase 7 (ENTPD7). Functional assays demonstrated that gga-miR-6660-3p inhibits myoblast proliferation and differentiation capacity while suppressing the de novo IMP synthesis pathway. However, circACLY reverses the inhibitory effects of gga-miR-6660-3p on these processes. This study systematically elucidates the regulatory mechanism of the circACLY/gga-miR-6660-3p/ENTPD7 axis in the development of quail myoblasts and IMP synthesis, providing a theoretical basis for deepening the understanding of the molecular basis of skeletal muscle development in local chicken breeds and the genetic improvement of meat quality traits.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"107 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145907675","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Melissa Parent,Christine Bone,Lee-Anne Huber,E James Squires
Boar taint, an unpleasant odor in heated pork, is caused primarily by androstenone accumulation in fat. Previous research demonstrated that biochar (BC), a charcoal-based adsorbent, can reduce androstenone accumulation in the fat when used as a dietary additive. The objective of this study was to compare the effectiveness of BC as a dietary treatment for boar taint in two genetic lines of boars from Alliance Genetics Canada (AGC) and Pig Improvement Company (PIC), and examine the influence of steroidogenic capacity on treatment response. All boars were fed a 5% BC diet for four weeks starting at a bodyweight (BW) of 75.25 ± 1.42 kg and 111.10 ± 0.86 days of age (DOA), followed by a two-week recovery period where BC was removed from the diet. Plasma androstenone, estrone sulfate (E1S), and growth performance parameters were measured weekly, and fat androstenone was measured biweekly. We found that 71% of PIC boars and 47% of AGC boars never developed boar taint, as the fat androstenone concentrations remained below the threshold of 1 μg/g throughout the trial. Of those with the capacity to develop boar taint, 89% of AGC boars responded to BC treatment, compared to 67% for PIC. However, no differences in plasma or fat androstenone levels were observed between the two genetic lines. Additionally, all growth performance parameters were not different between AGC and PIC boars except gain: feed (G: F) during the treatment period and average daily gain (ADG) during recovery, where PIC boars performed better (P = 0.001). Plasma E1S concentrations measured at approximately 130 kg live weight were used to classify boars as either high (plasma E1S > 16 ng/mL) or low steroidogenic capacity (plasma E1S ≤ 16 ng/mL). High steroidogenic capacity boars had greater plasma E1S concentrations (P < 0.05) across all time points compared those with low steroidogenic capacity, although growth rates remained not different between groups. In both genetic lines, boars that never developed boar taint as well as those responded to BC treatment consisted of a mix of high and low steroidogenic capacity animals, while boars that did not respond to BC treatment were exclusively high steroidogenic capacity. These results suggest that steroidogenic capacity may help to predict BC treatment response, although additional research is needed to identify genetic and biological markers associated with treatment success in order to effectively tailor BC treatment to animals most likely to benefit.
{"title":"Effect of Genetic Line and Steroidogenic Capacity on Response to Dietary Supplementation of Biochar to Control Fat Androstenone Levels in Boars.","authors":"Melissa Parent,Christine Bone,Lee-Anne Huber,E James Squires","doi":"10.1093/jas/skaf463","DOIUrl":"https://doi.org/10.1093/jas/skaf463","url":null,"abstract":"Boar taint, an unpleasant odor in heated pork, is caused primarily by androstenone accumulation in fat. Previous research demonstrated that biochar (BC), a charcoal-based adsorbent, can reduce androstenone accumulation in the fat when used as a dietary additive. The objective of this study was to compare the effectiveness of BC as a dietary treatment for boar taint in two genetic lines of boars from Alliance Genetics Canada (AGC) and Pig Improvement Company (PIC), and examine the influence of steroidogenic capacity on treatment response. All boars were fed a 5% BC diet for four weeks starting at a bodyweight (BW) of 75.25 ± 1.42 kg and 111.10 ± 0.86 days of age (DOA), followed by a two-week recovery period where BC was removed from the diet. Plasma androstenone, estrone sulfate (E1S), and growth performance parameters were measured weekly, and fat androstenone was measured biweekly. We found that 71% of PIC boars and 47% of AGC boars never developed boar taint, as the fat androstenone concentrations remained below the threshold of 1 μg/g throughout the trial. Of those with the capacity to develop boar taint, 89% of AGC boars responded to BC treatment, compared to 67% for PIC. However, no differences in plasma or fat androstenone levels were observed between the two genetic lines. Additionally, all growth performance parameters were not different between AGC and PIC boars except gain: feed (G: F) during the treatment period and average daily gain (ADG) during recovery, where PIC boars performed better (P = 0.001). Plasma E1S concentrations measured at approximately 130 kg live weight were used to classify boars as either high (plasma E1S > 16 ng/mL) or low steroidogenic capacity (plasma E1S ≤ 16 ng/mL). High steroidogenic capacity boars had greater plasma E1S concentrations (P < 0.05) across all time points compared those with low steroidogenic capacity, although growth rates remained not different between groups. In both genetic lines, boars that never developed boar taint as well as those responded to BC treatment consisted of a mix of high and low steroidogenic capacity animals, while boars that did not respond to BC treatment were exclusively high steroidogenic capacity. These results suggest that steroidogenic capacity may help to predict BC treatment response, although additional research is needed to identify genetic and biological markers associated with treatment success in order to effectively tailor BC treatment to animals most likely to benefit.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"24 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145907926","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Rodrigo D L Pacheco,Jéssica O Gusmão,Alex S Maia,Pedro H P Terêncio,João P S T Bastos,Ricardo Ávila,Bradley J Johnson,Victor P C Campanelli,Vinícius F C Fonsêca,Gustavo A B Moura,Rafael Da C Cervieri,John C Fuller
β-Hydroxy-β-methylbutyrate (HMB), a metabolite of leucine, increases both protein synthesis and lipolysis, immune function, and overall health in both humans and animals. Previous cattle studies have shown that HMB, fed as calcium HMB (CaHMB), can minimize stress responses and improve meat quality. The objective of the present study was to investigate the effect of feeding CaHMB in a subtropical feedlot on performance of Nellore (Bos indicus) bulls. A total of 3,520 bulls (64 pens; 55 animals per pen) were studied with four levels of CaHMB (0 (control, CON), 3, 5, or 7 g per head per day) and half the pens had shade access (SH), and half did not in a split-plot experimental design. Diets containing CaHMB were fed during three phases of the study, adaptation, growth, and finishing. There were no CaHMB x SH interactions observed for any variable measured. However, adjusted final body weight (FBW) tended to increase in CaHMB-treated bulls up to 5.8 kg in bulls fed 3 g per day (linear P < 0.07). Additionally, adjusted average daily gain (ADG) and carcass ADG were also increased with 3 g per day CaHMB by 3.2% and 2.6%, respectively, when compared with CON (linear P < 0.04). Bulls fed CaHMB tended to have increased hot carcass weight (HCW) by up to 3.4 kg in bulls fed 3 g CaHMB per day and a 0.36% increase in dressing percentage (DP) (linear P < 0.07 and 0.09, respectively). No differences were observed in dry matter intake (DMI) during any of the feeding periods. No significant differences were observed in gain: feed. Of major importance to the subtropical environment, 34% of the days were considered hot to very hot with the comfort level (InCI) >0.45 on a 0 to 1.0 scale. In conclusion, supplementing Bos indicus bulls with CaHMB resulted in improved growth performance and carcass yield.
β-羟基-β-甲基丁酸盐（HMB）是亮氨酸的代谢物，可促进蛋白质合成和脂肪分解、免疫功能以及人类和动物的整体健康。以前的牛研究表明，以钙HMB （CaHMB）喂养的HMB可以减少应激反应并改善肉品质。本研究旨在探讨在亚热带饲养场饲喂CaHMB对牛生产性能的影响。共有3,520头公牛（64个栏，每个栏55头动物）在四种水平的CaHMB（0（对照，CON）， 3,5或7 g /头/天）下进行了研究，一半的栏有荫蔽通道（SH），一半没有。在适应期、生长期和肥育期三个阶段分别饲喂含有CaHMB的饲粮。在测量的任何变量中，没有观察到CaHMB与SH的相互作用。然而，在每天饲喂3 g cahmb的公牛中，调整后的最终体重（FBW）倾向于增加5.8 kg（在0 ~ 1.0范围内线性P为0.45）。综上所述，在公牛中添加CaHMB可提高公牛的生长性能和胴体产量。
{"title":"Evaluation of calcium β-hydroxy-β-methylbutyrate on performance of Bos indicus-influenced cattle in a subtropical environment.","authors":"Rodrigo D L Pacheco,Jéssica O Gusmão,Alex S Maia,Pedro H P Terêncio,João P S T Bastos,Ricardo Ávila,Bradley J Johnson,Victor P C Campanelli,Vinícius F C Fonsêca,Gustavo A B Moura,Rafael Da C Cervieri,John C Fuller","doi":"10.1093/jas/skaf459","DOIUrl":"https://doi.org/10.1093/jas/skaf459","url":null,"abstract":"β-Hydroxy-β-methylbutyrate (HMB), a metabolite of leucine, increases both protein synthesis and lipolysis, immune function, and overall health in both humans and animals. Previous cattle studies have shown that HMB, fed as calcium HMB (CaHMB), can minimize stress responses and improve meat quality. The objective of the present study was to investigate the effect of feeding CaHMB in a subtropical feedlot on performance of Nellore (Bos indicus) bulls. A total of 3,520 bulls (64 pens; 55 animals per pen) were studied with four levels of CaHMB (0 (control, CON), 3, 5, or 7 g per head per day) and half the pens had shade access (SH), and half did not in a split-plot experimental design. Diets containing CaHMB were fed during three phases of the study, adaptation, growth, and finishing. There were no CaHMB x SH interactions observed for any variable measured. However, adjusted final body weight (FBW) tended to increase in CaHMB-treated bulls up to 5.8 kg in bulls fed 3 g per day (linear P < 0.07). Additionally, adjusted average daily gain (ADG) and carcass ADG were also increased with 3 g per day CaHMB by 3.2% and 2.6%, respectively, when compared with CON (linear P < 0.04). Bulls fed CaHMB tended to have increased hot carcass weight (HCW) by up to 3.4 kg in bulls fed 3 g CaHMB per day and a 0.36% increase in dressing percentage (DP) (linear P < 0.07 and 0.09, respectively). No differences were observed in dry matter intake (DMI) during any of the feeding periods. No significant differences were observed in gain: feed. Of major importance to the subtropical environment, 34% of the days were considered hot to very hot with the comfort level (InCI) >0.45 on a 0 to 1.0 scale. In conclusion, supplementing Bos indicus bulls with CaHMB resulted in improved growth performance and carcass yield.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"52 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145907716","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
No studies are currently available regarding the roles of the amino acid transporters in promoting mineral absorption in animals. To study whether the y(+)L-type amino acid transporter 2 (y+LAT2) participates in enhancing Zn absorption as Zn proteinate with moderate chelation strength (Zn-Prot M) in broilers, three experiments were carried out to investigate the effects of y+LAT2 silencing and overexpression on zinc (Zn) absorption as Zn-Prot M and related gene expressions in broiler primary duodenal epithelial cells (BPDECs) contrasted with Zn sulfate (ZnSO4). In Exp.1, three pairs of siRNAs (si-494, si-834 and si-1153) targeting y+LAT2 were synthesized and transfected within the cells to identify the optimal siRNA for silencing y+LAT2 mRNA expression, and the data were analyzed by one-way ANOVA. In Exp.2, cells were incubated for 2 h in either a DMEM/F12 complete medium without Zn supplementation (Control) or the same medium enriched with 400 μmol Zn/L as ZnSO4 or Zn-Prot M, with or without the only one effective siRNA targeting y+LAT2, and the data were analyzed by two-way ANOVA. In Exp.3, with or without the recombinant adenovirus plasmid encoding y+LAT2, cells were also treated for 2 h with the same media as in Exp. 2, and the data were analyzed by two-way ANOVA. The results demonstrated that in Exp.1, the si-494 was the only one effective (P < 0.0001) siRNA for inhibiting y+LAT2 mRNA expression. In Exp.2, y+LAT2 silencing reduced its own mRNA and protein abundances (P < 0.0001) by 46% and 14% and Zn absorption from Zn-Prot M by 34% (P < 0.05), with no effect (P > 0.05) on Zn absorption from ZnSO4. In Exp.3, y+LAT2 overexpression elevated its own mRNA and protein expressions by 949% and 18% (P < 0.0001) and Zn absorption from Zn-Prot M by 73% (P < 0.05), but didn't affect (P > 0.05) Zn absorption from ZnSO4. Additionally, Zn-Prot M upregulated (P < 0.05) the y+LAT2 protein expression compared to the Control and ZnSO4 by 12∼17%, with no impact (P > 0.07) on the y+LAT2 mRNA expression, regardless of y+LAT2 silencing or overexpression. In conclusion, the y+LAT2 took part in promoting Zn absorption as Zn-Prot M in BPDECs.
{"title":"The role of the y(+)L-type amino acid transporter 2 in promoting zinc absorption as zinc proteinate with moderate chelation strength in broiler primary duodenal epithelial cells.","authors":"Chunyu Cao,Yun Hu,Liang Huang,Weiyun Zhang,Xi Lin,Wei Wu,Tingting Li,Xiaoyan Cui,Shengchen Wang,Liyang Zhang,Xugang Luo","doi":"10.1093/jas/skaf466","DOIUrl":"https://doi.org/10.1093/jas/skaf466","url":null,"abstract":"No studies are currently available regarding the roles of the amino acid transporters in promoting mineral absorption in animals. To study whether the y(+)L-type amino acid transporter 2 (y+LAT2) participates in enhancing Zn absorption as Zn proteinate with moderate chelation strength (Zn-Prot M) in broilers, three experiments were carried out to investigate the effects of y+LAT2 silencing and overexpression on zinc (Zn) absorption as Zn-Prot M and related gene expressions in broiler primary duodenal epithelial cells (BPDECs) contrasted with Zn sulfate (ZnSO4). In Exp.1, three pairs of siRNAs (si-494, si-834 and si-1153) targeting y+LAT2 were synthesized and transfected within the cells to identify the optimal siRNA for silencing y+LAT2 mRNA expression, and the data were analyzed by one-way ANOVA. In Exp.2, cells were incubated for 2 h in either a DMEM/F12 complete medium without Zn supplementation (Control) or the same medium enriched with 400 μmol Zn/L as ZnSO4 or Zn-Prot M, with or without the only one effective siRNA targeting y+LAT2, and the data were analyzed by two-way ANOVA. In Exp.3, with or without the recombinant adenovirus plasmid encoding y+LAT2, cells were also treated for 2 h with the same media as in Exp. 2, and the data were analyzed by two-way ANOVA. The results demonstrated that in Exp.1, the si-494 was the only one effective (P < 0.0001) siRNA for inhibiting y+LAT2 mRNA expression. In Exp.2, y+LAT2 silencing reduced its own mRNA and protein abundances (P < 0.0001) by 46% and 14% and Zn absorption from Zn-Prot M by 34% (P < 0.05), with no effect (P > 0.05) on Zn absorption from ZnSO4. In Exp.3, y+LAT2 overexpression elevated its own mRNA and protein expressions by 949% and 18% (P < 0.0001) and Zn absorption from Zn-Prot M by 73% (P < 0.05), but didn't affect (P > 0.05) Zn absorption from ZnSO4. Additionally, Zn-Prot M upregulated (P < 0.05) the y+LAT2 protein expression compared to the Control and ZnSO4 by 12∼17%, with no impact (P > 0.07) on the y+LAT2 mRNA expression, regardless of y+LAT2 silencing or overexpression. In conclusion, the y+LAT2 took part in promoting Zn absorption as Zn-Prot M in BPDECs.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"42 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145907672","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The effect of graded reductions in balanced crude protein (CP) on hen productive performance, egg quality, nitrogen balance, abdominal fat deposition, tibia traits, and relative economic outcomes from 26 to 44 weeks of age (WOA) was investigated. A total of 252 26-week-old Hy-Line Brown hens were housed in enriched cages (7 birds/cage) and randomly allocated to 1 of 4 different dietary CP levels of iso-energetic diets with 9 replicates per treatment. The trial was conducted over two phases of 26 to 34 and 36 to 44 WOA. Diets included a high-protein (HP; 18.0% and 17.0% CP in Phases 1 and 2), medium-protein (MP), low-protein (LP), and very low-protein (VLP) series, representing stepwise reductions of 0.50, 1.00, and 1.50 percentage points relative to HP. Limiting amino acids (AA; lysine, methionine, and threonine) were supplemented to ensure balanced AA profiles. Collected data were analyzed using the General Linear Model (GLM) procedure for one-way ANOVA; statistical significance was set at P < 0.05, and trends were noted at 0.05 < P < 0.10. Balanced protein reduction tended to improve abdominal fat contents (2.66 to 2.85%; P = 0.059), but reduce body weight gain (141.66 to 95.66 g; P = 0.089), particularly with the VLP diet. Across 26 to 44 WOA, graded CP reduction lowered egg weight (60.36 to 59.40 g; P < 0.05) and feed conversion efficiency (1.93 to 1.97 g feed/g egg; P < 0.05); and tended to reduce egg mass (57.02 to 55.11 g/hen/day; P = 0.080), particularly in the VLP group. As to egg quality, Haugh units were higher (P < 0.05) with HP and MP diets (94.60 and 94.30) than LP and VLP diets (93.66 and 93.04) across 26-44 WOA. In contrast, LP and VLP diets tended to improve yolk color (8.38 to 8.49; P = 0.076) and egg-breaking strength (5.39 to 5.51 kg; P = 0.058) across 26-44 WOA. Dietary CP reduction linearly reduced nitrogen consumed and excreted by more than 10% (P < 0.05). Tibia-breaking strength tended to decline with dietary CP reduction (P = 0.094), decreasing from 27.62 kg in HP to 25.54-25.68 kg in the LP and VLP diets. Economically, reduced CP lowered egg income (P < 0.05) at weeks 34 and 44 (2.00 to 1.77$; 1.96 to 1.89$, respectively); and feed costs at week 34 only (0.54 to 0.52$; P = 0.088), but profit margins remained unaffected (P > 0.10). Conclusively, these results confirm the effectiveness of balanced dietary protein reduction in maintaining egg production rate and most egg quality traits while reducing nitrogen excreted and feed costs.
{"title":"Laying hen responses to balanced protein reduction on performance, egg quality, nitrogen balance, and fat and mineral utilization.","authors":"Elijah Ogola Oketch,Myunghwan Yu,Shan Randima Nawarathne,Nuwan Chamara Chathuranga,Jeseok Lee,Haeeun Park,Bo Keun Lee,Kwan Eung Kim,Jung Min Heo","doi":"10.1093/jas/skaf465","DOIUrl":"https://doi.org/10.1093/jas/skaf465","url":null,"abstract":"The effect of graded reductions in balanced crude protein (CP) on hen productive performance, egg quality, nitrogen balance, abdominal fat deposition, tibia traits, and relative economic outcomes from 26 to 44 weeks of age (WOA) was investigated. A total of 252 26-week-old Hy-Line Brown hens were housed in enriched cages (7 birds/cage) and randomly allocated to 1 of 4 different dietary CP levels of iso-energetic diets with 9 replicates per treatment. The trial was conducted over two phases of 26 to 34 and 36 to 44 WOA. Diets included a high-protein (HP; 18.0% and 17.0% CP in Phases 1 and 2), medium-protein (MP), low-protein (LP), and very low-protein (VLP) series, representing stepwise reductions of 0.50, 1.00, and 1.50 percentage points relative to HP. Limiting amino acids (AA; lysine, methionine, and threonine) were supplemented to ensure balanced AA profiles. Collected data were analyzed using the General Linear Model (GLM) procedure for one-way ANOVA; statistical significance was set at P < 0.05, and trends were noted at 0.05 < P < 0.10. Balanced protein reduction tended to improve abdominal fat contents (2.66 to 2.85%; P = 0.059), but reduce body weight gain (141.66 to 95.66 g; P = 0.089), particularly with the VLP diet. Across 26 to 44 WOA, graded CP reduction lowered egg weight (60.36 to 59.40 g; P < 0.05) and feed conversion efficiency (1.93 to 1.97 g feed/g egg; P < 0.05); and tended to reduce egg mass (57.02 to 55.11 g/hen/day; P = 0.080), particularly in the VLP group. As to egg quality, Haugh units were higher (P < 0.05) with HP and MP diets (94.60 and 94.30) than LP and VLP diets (93.66 and 93.04) across 26-44 WOA. In contrast, LP and VLP diets tended to improve yolk color (8.38 to 8.49; P = 0.076) and egg-breaking strength (5.39 to 5.51 kg; P = 0.058) across 26-44 WOA. Dietary CP reduction linearly reduced nitrogen consumed and excreted by more than 10% (P < 0.05). Tibia-breaking strength tended to decline with dietary CP reduction (P = 0.094), decreasing from 27.62 kg in HP to 25.54-25.68 kg in the LP and VLP diets. Economically, reduced CP lowered egg income (P < 0.05) at weeks 34 and 44 (2.00 to 1.77$; 1.96 to 1.89$, respectively); and feed costs at week 34 only (0.54 to 0.52$; P = 0.088), but profit margins remained unaffected (P > 0.10). Conclusively, these results confirm the effectiveness of balanced dietary protein reduction in maintaining egg production rate and most egg quality traits while reducing nitrogen excreted and feed costs.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"11 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145907680","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The luteinizing hormone (LH) is well established to trigger oocyte maturation and cumulus expansion in preovulatory follicles, primarily through activating the epidermal growth factor (EGF) signaling network. While previous studies have demonstrated that activation of the G protein-coupled estrogen receptor (GPR30) accelerates meiotic resumption and first polar body extrusion in oocytes, the molecular mechanisms underlying GPR30 function in this process remain poorly defined. Here, cumulus-oocyte complexes (COCs) of goat follicles were used to investigate the interaction between the EGF network and GPR30 during oocyte maturation and ovulation. Our results showed that: LH signal was found to increase GPR30 protein levels via the EGF receptor signaling pathway (P＜0.05); Estradiol (E2) acts via GPR30 to promote ERK1/2 phosphorylation, cyclic adenosine monophosphate (cAMP) production, first polar body extrusion, and the expression of cumulus expansion-related genes in COCs (P＜0.05), and E2 further enhances these functional outcomes by increasing EGFR protein levels in cumulus cells. Collectively, these findings reveal a synergistic interaction between LH and E2 in regulating goat oocyte maturation: E2, via GPR30-mediated upregulation of EGFR, enhances COCs' responsiveness to LH-induced EGF signaling-thereby amplifying oocyte maturation efficiency and cumulus expansion. This study provides new insights into the integrated signaling network governing mammalian oocyte maturation and offers a potential molecular target for optimizing in vitro maturation (IVM) protocols for goat.
{"title":"Cooperation of luteinizing hormone and estradiol in the promotion of goat oocyte maturation via regulation and activation of the GPR30 and EGFR signaling pathways.","authors":"Baijuan Yue,Jie Liu,Sihai Lu,Meini Yu,Lele Zhu,Yaju Tang,Xiaoe Zhao,Sha Peng,Menghao Pan,Qiang Wei,Baohua Ma","doi":"10.1093/jas/skaf462","DOIUrl":"https://doi.org/10.1093/jas/skaf462","url":null,"abstract":"The luteinizing hormone (LH) is well established to trigger oocyte maturation and cumulus expansion in preovulatory follicles, primarily through activating the epidermal growth factor (EGF) signaling network. While previous studies have demonstrated that activation of the G protein-coupled estrogen receptor (GPR30) accelerates meiotic resumption and first polar body extrusion in oocytes, the molecular mechanisms underlying GPR30 function in this process remain poorly defined. Here, cumulus-oocyte complexes (COCs) of goat follicles were used to investigate the interaction between the EGF network and GPR30 during oocyte maturation and ovulation. Our results showed that: LH signal was found to increase GPR30 protein levels via the EGF receptor signaling pathway (P＜0.05); Estradiol (E2) acts via GPR30 to promote ERK1/2 phosphorylation, cyclic adenosine monophosphate (cAMP) production, first polar body extrusion, and the expression of cumulus expansion-related genes in COCs (P＜0.05), and E2 further enhances these functional outcomes by increasing EGFR protein levels in cumulus cells. Collectively, these findings reveal a synergistic interaction between LH and E2 in regulating goat oocyte maturation: E2, via GPR30-mediated upregulation of EGFR, enhances COCs' responsiveness to LH-induced EGF signaling-thereby amplifying oocyte maturation efficiency and cumulus expansion. This study provides new insights into the integrated signaling network governing mammalian oocyte maturation and offers a potential molecular target for optimizing in vitro maturation (IVM) protocols for goat.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"177 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145907674","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Germán D Ramírez-Zamudio, Wellison J S Diniz, Jennifer L Hurlbert, Priyanka Banerjee, Roberta B A Dahlen, Ana Clara B Menezes, Kendall C Swanson, Carl R Dahlen
Supplementation with vitamins and minerals (VTM) in the diet of pregnant F0 dams influences offspring development; however, long-term effects on metabolic organs, including the liver, remain unclear. This study evaluated hepatic gene expression in F1 heifers born to F0 dams fed a basal diet with VTM (n = 8) or without VTM (CON; n = 7) throughout gestation. After birth, F1 heifers were managed as a single group and inseminated with sexed female semen from a single sire. At 250 days of gestation and ∼22 months of age, F1 heifers were harvested and liver samples collected. Total RNA was extracted, sequenced, and reads aligned using STAR after quality control. Differential expression analyses were performed in R (v4.4.2) using the DESeq2 package, and over-representation of KEGG pathways and biological processes was carried out using ShinyGO v.0.76. A total of 13,990 genes were tested, of which 486 were differentially expressed (P ≤ 0.05; |log2FC| ≥ 0.5), including 241 upregulated and 245 downregulated in the VTM group. Upregulated genes by maternal VTM supplementation included RPL23, SOX4, PTGS2, NDUFC1, UQCRH, and RPS28, associated with pathways including ribosome and processes like cell proliferation and oxidative phosphorylation. Downregulated genes, including EIF2AK2, ESR1, SLC7A1, and PIK3CB, were associated with processes related to regulation of cell population proliferation and organonitrogen compound biosynthetic process. These results indicate that VTM supplementation in F0 dams may induce persistent changes in hepatic gene expression in the offspring, with potential metabolic implications during critical physiological stages, such as during gestation in F1 heifers. (Supported by USDA 2022-67016-36479)
{"title":"45 Vitamin and mineral supplementation in pregnant F0 beef dams may induce persistent changes in the hepatic transcriptome of F1 heifers","authors":"Germán D Ramírez-Zamudio, Wellison J S Diniz, Jennifer L Hurlbert, Priyanka Banerjee, Roberta B A Dahlen, Ana Clara B Menezes, Kendall C Swanson, Carl R Dahlen","doi":"10.1093/jas/skaf398.038","DOIUrl":"https://doi.org/10.1093/jas/skaf398.038","url":null,"abstract":"Supplementation with vitamins and minerals (VTM) in the diet of pregnant F0 dams influences offspring development; however, long-term effects on metabolic organs, including the liver, remain unclear. This study evaluated hepatic gene expression in F1 heifers born to F0 dams fed a basal diet with VTM (n = 8) or without VTM (CON; n = 7) throughout gestation. After birth, F1 heifers were managed as a single group and inseminated with sexed female semen from a single sire. At 250 days of gestation and ∼22 months of age, F1 heifers were harvested and liver samples collected. Total RNA was extracted, sequenced, and reads aligned using STAR after quality control. Differential expression analyses were performed in R (v4.4.2) using the DESeq2 package, and over-representation of KEGG pathways and biological processes was carried out using ShinyGO v.0.76. A total of 13,990 genes were tested, of which 486 were differentially expressed (P ≤ 0.05; |log2FC| ≥ 0.5), including 241 upregulated and 245 downregulated in the VTM group. Upregulated genes by maternal VTM supplementation included RPL23, SOX4, PTGS2, NDUFC1, UQCRH, and RPS28, associated with pathways including ribosome and processes like cell proliferation and oxidative phosphorylation. Downregulated genes, including EIF2AK2, ESR1, SLC7A1, and PIK3CB, were associated with processes related to regulation of cell population proliferation and organonitrogen compound biosynthetic process. These results indicate that VTM supplementation in F0 dams may induce persistent changes in hepatic gene expression in the offspring, with potential metabolic implications during critical physiological stages, such as during gestation in F1 heifers. (Supported by USDA 2022-67016-36479)","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"5 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145894367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bradley Lawrence, Roberto Barea, Marisol Castillo, Silvia Peris, Jesus Acosta, Xandra Benthem de Grave, Francesc Molist, Deana Hancock
: Introduction Offspring from sows fed Zn, Cu and Mn in the metal methionine hydroxy analogue chelate (MHAC) form vs. sulfate form have been shown to have greater loin eye area at harvest. Data is lacking compared with other mineral sources. Objective Assess the impact of maternal feeding of MHAC or glycinate (Gly) minerals on offspring serum mineral concentration, loin depth and fat depth. Methods From 6 weeks pre-breeding through first gestation, diets were supplemented with 80 mg/kg Zn, 10 mg/kg Cu and 20 mg/kg Mn. From first through third lactation, Zn, Cu and Mn were provided at 50 mg/kg Zn, 10 mg/kg Cu and 20 mg/kg Mn. During parity 3, serum from 1 piglet/litter at 4-d, weaning and 14-d post-weaning was obtained from 20 litters/treatment. At each parity, muscle and backfat depth was determined for approximately 1,000 pigs/treatment. Results There was no treatment (P > 0.15) nor treatment × time interaction (P > 0.22) for serum mineral concentration. Serum Fe, Zn, Cu and Mn were highest at weaning (P < 0.001). Cu and Zn concentrations were 34% and 60% lower respectively at 14-d post-weaning than at 4-d of age. Parity 2 and 3 MHAC offspring had 0.5 mm (P < 0.01) and 1.0 mm (P < 0.001) more muscle depth at 122 kg. Backfat was lower in parity 1 (-0.2 mm; P < 0.01) and parity 3 (-0.4 mm; P < 0.001) MHAC offspring. Conclusions Offspring serum mineral concentration was highest at 4-d of age. Maternal trace mineral source appears to provide an opportunity to impact offspring loin and fat depth at harvest.
{"title":"29 Maternal mineral source impact on piglet serum mineral and carcass composition at harvest","authors":"Bradley Lawrence, Roberto Barea, Marisol Castillo, Silvia Peris, Jesus Acosta, Xandra Benthem de Grave, Francesc Molist, Deana Hancock","doi":"10.1093/jas/skaf398.023","DOIUrl":"https://doi.org/10.1093/jas/skaf398.023","url":null,"abstract":": Introduction Offspring from sows fed Zn, Cu and Mn in the metal methionine hydroxy analogue chelate (MHAC) form vs. sulfate form have been shown to have greater loin eye area at harvest. Data is lacking compared with other mineral sources. Objective Assess the impact of maternal feeding of MHAC or glycinate (Gly) minerals on offspring serum mineral concentration, loin depth and fat depth. Methods From 6 weeks pre-breeding through first gestation, diets were supplemented with 80 mg/kg Zn, 10 mg/kg Cu and 20 mg/kg Mn. From first through third lactation, Zn, Cu and Mn were provided at 50 mg/kg Zn, 10 mg/kg Cu and 20 mg/kg Mn. During parity 3, serum from 1 piglet/litter at 4-d, weaning and 14-d post-weaning was obtained from 20 litters/treatment. At each parity, muscle and backfat depth was determined for approximately 1,000 pigs/treatment. Results There was no treatment (P &gt; 0.15) nor treatment × time interaction (P &gt; 0.22) for serum mineral concentration. Serum Fe, Zn, Cu and Mn were highest at weaning (P &lt; 0.001). Cu and Zn concentrations were 34% and 60% lower respectively at 14-d post-weaning than at 4-d of age. Parity 2 and 3 MHAC offspring had 0.5 mm (P &lt; 0.01) and 1.0 mm (P &lt; 0.001) more muscle depth at 122 kg. Backfat was lower in parity 1 (-0.2 mm; P &lt; 0.01) and parity 3 (-0.4 mm; P &lt; 0.001) MHAC offspring. Conclusions Offspring serum mineral concentration was highest at 4-d of age. Maternal trace mineral source appears to provide an opportunity to impact offspring loin and fat depth at harvest.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"15 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145847360","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lisa A Vrooman, Josue Baeza, Ana C Lima, Eric Rhon-Calderon, Donald F Conrad, Benjamin A Garcia, Marisa S Bartolomei
Pregnancies utilizing Assisted Reproductive Technologies (ART) are associated with several complications including an increased risk of preeclampsia, placental abruption, and morbidly adherent placentas. Our work along with others has shown that placentas from mice conceived by in vitro fertilization (IVF) display overgrowth, impaired blood vessel development, altered gene expression and DNA methylation. These changes are associated with impaired fetal growth. We sought to identify the placental protein differences with IVF to inform potential interventions for improving ART procedures. Placental protein differences among IVF and spontaneously-conceived mouse concepti were analyzed in a sex-specific manner (n = 4-5 placentas/sex/experimental group/timepoint) by data-independent acquisition mass spectrometry at five timepoints covering shortly after placental formation through term (E11.5, 12.5, 14.5, 16.5, and 18.5). Peptides below 1% FDR were included, and statistical analysis was performed using Two-way ANOVA and Tukey HSD post hoc tests, and adjustment for multiple hypothesis testing. We observed normal, dynamic abundance changes for placental proteins over development. IVF induces an overall reduction in the abundance of several placental proteins at the earliest timepoint, E11.5. Several of the identified affected proteins are known to be important for placenta development and epigenetic regulation. Intriguingly, we observed most placental protein changes were observed among the female concepti but not the male concepti. To our knowledge, this is the first proteomic analysis of mouse placentas at multiple timepoints along gestation. We are currently conducting spatial transcriptomics experiments to determine if protein changes are associated with specific placental compartments.
{"title":"60 Investigating placental protein changes associated with Assisted Reproductive Technologies in a mouse model","authors":"Lisa A Vrooman, Josue Baeza, Ana C Lima, Eric Rhon-Calderon, Donald F Conrad, Benjamin A Garcia, Marisa S Bartolomei","doi":"10.1093/jas/skaf398.051","DOIUrl":"https://doi.org/10.1093/jas/skaf398.051","url":null,"abstract":"Pregnancies utilizing Assisted Reproductive Technologies (ART) are associated with several complications including an increased risk of preeclampsia, placental abruption, and morbidly adherent placentas. Our work along with others has shown that placentas from mice conceived by in vitro fertilization (IVF) display overgrowth, impaired blood vessel development, altered gene expression and DNA methylation. These changes are associated with impaired fetal growth. We sought to identify the placental protein differences with IVF to inform potential interventions for improving ART procedures. Placental protein differences among IVF and spontaneously-conceived mouse concepti were analyzed in a sex-specific manner (n = 4-5 placentas/sex/experimental group/timepoint) by data-independent acquisition mass spectrometry at five timepoints covering shortly after placental formation through term (E11.5, 12.5, 14.5, 16.5, and 18.5). Peptides below 1% FDR were included, and statistical analysis was performed using Two-way ANOVA and Tukey HSD post hoc tests, and adjustment for multiple hypothesis testing. We observed normal, dynamic abundance changes for placental proteins over development. IVF induces an overall reduction in the abundance of several placental proteins at the earliest timepoint, E11.5. Several of the identified affected proteins are known to be important for placenta development and epigenetic regulation. Intriguingly, we observed most placental protein changes were observed among the female concepti but not the male concepti. To our knowledge, this is the first proteomic analysis of mouse placentas at multiple timepoints along gestation. We are currently conducting spatial transcriptomics experiments to determine if protein changes are associated with specific placental compartments.","PeriodicalId":14895,"journal":{"name":"Journal of animal science","volume":"31 1","pages":""},"PeriodicalIF":3.3,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145847369","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：