Excess ammonia(NH_(3))in the circulation of dairy animals can reduce animal health and the quality of products for human consumption.To develop effective prevention and treatment methods,it is essential to examine the...Excess ammonia(NH_(3))in the circulation of dairy animals can reduce animal health and the quality of products for human consumption.To develop effective prevention and treatment methods,it is essential to examine the molecular mechanisms through which excess NH_(3) may affect the mammary gland.The present study used bovine mammary epithelial cells(BMECs)to evaluate the effects of exogenous NH_(4)Cl on the abundance of circular RNAs(circRNAs)using high-throughput sequencing.Among the identified circRNAs,circ02771 was the most significantly upregulated by exogenous NH_(4)Cl(P<0.05),with a fold change of 4.12.The results of the apoptosis and proliferation assays,transmission electron microscopy,H&E staining,and immunohistochemistry revealed that circ02771 increased apoptosis and inflammation.A double luciferase reporter assay revealed that circ02771 targeted miR-194b,and the overexpression of circ02771(pcDNA-circ02771)reduced(P<0.05)the expression of miR-194b and led to apoptosis and inflammation.Circ02771 also enhanced the expression of transforming growth factor beta-induced factor homeobox 1(TGIF1),which is a target gene of miR-194b.Overall,this study suggests that the circ02771/miR-194b/TGIF1 axis plays a role in mediating the effects of NH_(4)Cl on BMECs.Therefore,this axis provides a novel target to help control hazards within the mammary gland from high circulating NH_(4)Cl levels.展开更多
Batch cultures of mixed rumen micro-organisms were conducted to evaluate the effects of encapsulated yeast(+EY)and encapsulated enzyme(+EE)using plant proteins(barley and oats grain)on rumen fermentation in vitro,inve...Batch cultures of mixed rumen micro-organisms were conducted to evaluate the effects of encapsulated yeast(+EY)and encapsulated enzyme(+EE)using plant proteins(barley and oats grain)on rumen fermentation in vitro,investigate the abilities of encapsulated yeast and encapsulated enzyme to prevent rumen digestion in vitro.Treatments of the study were the control,+EY,+EE products(3.33 mg·mL^(-1) of the incubation medium),unencapsulated yeast(-EY)and enzyme(-EE)products(0.17 and 0.17μL·mL^(-1) of the incubation medium,respectively).+EY group increased dry matter disappearance(DMD,P<0.01)and the total volatile fatty acids(TVFA,P<0.01)at 3 h of the incubation compared with the control,regardless of encapsulation of yeast.Gas production(GP)of+EY group was higher(P=0.05,29.94 mL·mL^(-1) organic matter,OM)than that of the control(25.08 mL·g^(-1) OM)at 3 h of the incubation.Supplementation+EY increased DMD(P=0.04,0.394 vs 0.352,respectively)and acetic proportion(P=0.04,52.6 vs 49.8 mol•100 mL^(-1),respectively)at 6 h of the incubation and increased A:P ratio(P<0.01,3.11 and 2.86,respectively)at 24 h of the incubation,as compared to unencapsulation of yeast.Supplementation of enzyme had higher(P≤0.04)GP,DMD and TVFA at 3 and 6 h of the incubation compared with the control,regardless of encapsulation.Moreover,the addition of+EE produced greater GP at 6(P<0.01,92.35 vs 78.21 mL·g^(-1) OM,respectively),12(218.47 vs 159.18 mL·g^(-1) OM)and 24 h(380.97 vs 297.78 mL·g^(-1) OM,respectively)of the incubation,higher DMD(0.347 vs 0.313,respectively)at 3 h of the incubation as compared to-EE group.The study showed that the encapsulation might protect part of yeast and enzyme from releasing to the rumen throughout the digestion in vitro,resulting in higher or no difference of rumen fermentation parameters compared with unencapsulated groups at any incubation times.In comparison with-EY and-EE,the higher rumen fermentation parameters at the early incubation time were observed,which could be attributed to the higher concentration of yeast or enzyme.However,regardless of the encapsulation,the results indicated that both yeast and enzyme only improved the speed rather than the extent of rumen fermentation in vitro.展开更多
基金supported by the Independent Innovation in Jiangsu Province of China(CX(21)3119)the National Natural Science Foundation of China(31802035,31702095 and 31872324)+1 种基金the Jiangsu Natural Science Fund(BK20181221)Yangzhou Liangde Antibody Bio Tech.,China。
文摘Excess ammonia(NH_(3))in the circulation of dairy animals can reduce animal health and the quality of products for human consumption.To develop effective prevention and treatment methods,it is essential to examine the molecular mechanisms through which excess NH_(3) may affect the mammary gland.The present study used bovine mammary epithelial cells(BMECs)to evaluate the effects of exogenous NH_(4)Cl on the abundance of circular RNAs(circRNAs)using high-throughput sequencing.Among the identified circRNAs,circ02771 was the most significantly upregulated by exogenous NH_(4)Cl(P<0.05),with a fold change of 4.12.The results of the apoptosis and proliferation assays,transmission electron microscopy,H&E staining,and immunohistochemistry revealed that circ02771 increased apoptosis and inflammation.A double luciferase reporter assay revealed that circ02771 targeted miR-194b,and the overexpression of circ02771(pcDNA-circ02771)reduced(P<0.05)the expression of miR-194b and led to apoptosis and inflammation.Circ02771 also enhanced the expression of transforming growth factor beta-induced factor homeobox 1(TGIF1),which is a target gene of miR-194b.Overall,this study suggests that the circ02771/miR-194b/TGIF1 axis plays a role in mediating the effects of NH_(4)Cl on BMECs.Therefore,this axis provides a novel target to help control hazards within the mammary gland from high circulating NH_(4)Cl levels.
基金Supported by the Special Fund for Agro-scientific Research in the Public Interest(201503134)。
文摘Batch cultures of mixed rumen micro-organisms were conducted to evaluate the effects of encapsulated yeast(+EY)and encapsulated enzyme(+EE)using plant proteins(barley and oats grain)on rumen fermentation in vitro,investigate the abilities of encapsulated yeast and encapsulated enzyme to prevent rumen digestion in vitro.Treatments of the study were the control,+EY,+EE products(3.33 mg·mL^(-1) of the incubation medium),unencapsulated yeast(-EY)and enzyme(-EE)products(0.17 and 0.17μL·mL^(-1) of the incubation medium,respectively).+EY group increased dry matter disappearance(DMD,P<0.01)and the total volatile fatty acids(TVFA,P<0.01)at 3 h of the incubation compared with the control,regardless of encapsulation of yeast.Gas production(GP)of+EY group was higher(P=0.05,29.94 mL·mL^(-1) organic matter,OM)than that of the control(25.08 mL·g^(-1) OM)at 3 h of the incubation.Supplementation+EY increased DMD(P=0.04,0.394 vs 0.352,respectively)and acetic proportion(P=0.04,52.6 vs 49.8 mol•100 mL^(-1),respectively)at 6 h of the incubation and increased A:P ratio(P<0.01,3.11 and 2.86,respectively)at 24 h of the incubation,as compared to unencapsulation of yeast.Supplementation of enzyme had higher(P≤0.04)GP,DMD and TVFA at 3 and 6 h of the incubation compared with the control,regardless of encapsulation.Moreover,the addition of+EE produced greater GP at 6(P<0.01,92.35 vs 78.21 mL·g^(-1) OM,respectively),12(218.47 vs 159.18 mL·g^(-1) OM)and 24 h(380.97 vs 297.78 mL·g^(-1) OM,respectively)of the incubation,higher DMD(0.347 vs 0.313,respectively)at 3 h of the incubation as compared to-EE group.The study showed that the encapsulation might protect part of yeast and enzyme from releasing to the rumen throughout the digestion in vitro,resulting in higher or no difference of rumen fermentation parameters compared with unencapsulated groups at any incubation times.In comparison with-EY and-EE,the higher rumen fermentation parameters at the early incubation time were observed,which could be attributed to the higher concentration of yeast or enzyme.However,regardless of the encapsulation,the results indicated that both yeast and enzyme only improved the speed rather than the extent of rumen fermentation in vitro.
