[ Objective] The paper was to get effective soluble N protein to establish indirect ELISA method for Peste des petits ruminants （PPR）. [ Method] Soluble N protein with high expression was obtained from Escherichia c...[ Objective] The paper was to get effective soluble N protein to establish indirect ELISA method for Peste des petits ruminants （PPR）. [ Method] Soluble N protein with high expression was obtained from Escherichia coli expression system through codon optimization and optimization of expression conditions, and indirect ELISA detection method based on N protein was further established. [ Result] The assay had no cross reaction with other sheep pathogens. The intra- and inter-batch variation coefficients were less than 9%, indicating the method had good repeatability. Furthermore, totally 480 clinical serum samples were detec- ted by the assay, and the agreement rate with commercial ELISA kit （IDVET） was 98.33%. [ Conclusion] The study laid a foundation for further development of mature PPRV antibody detection kits.展开更多
[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PP...[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PPR.[Methods]Soluble N protein and NH fusion protein were successfully obtained in an Escherichia coli expression system by optimizing E.coli codon and expression conditions.Furthermore,based on purified soluble N protein and NH fusion protein,a double-antigen sandwich time-resolved fluorescence immunoassay method for detection of peste des petits ruminants virus(PPRV)was established.[Results]The method has high sensitivity and specificity and can specifically detect the antibody against PPRV in sheep serum,and it has no cross reaction with other related diseases.The method was used to detect 292 clinical samples,and compared with French IDVET competition ELISA kit.The coincidence rates of positive samples and negative samples from the two kinds of test kits were 92.47%and 97.26%,respectively,and the overall coincidence rate was 94.86%.The intra-group and inter-group coefficients of variation in the repeatability test were less than 10%.[Conclusions]Compared with the traditional ELISA method,the double-antigen sandwich time-resolved fluorescence immunoassay for detection of PPRV has equivalent sensitivity and specificity,and simple and rapid operation,and thus high application and popularization value.展开更多
Background： The use of antibiotics in animal diets is facing negative feedback due to the hidden danger of drug residues to human health. Traditional Chinese herbal medicine has been used to replace antibiotics in th...Background： The use of antibiotics in animal diets is facing negative feedback due to the hidden danger of drug residues to human health. Traditional Chinese herbal medicine has been used to replace antibiotics in the past two decades and played an increasingly important role in livestock production. The present study was carried out to assess the feeding effects of a traditional nourishing Chinese herbal medicine mixture on kinetics of plasma glucose, protein and energy metabolism in sheep. Ruminal fermentation characteristics were also determined. Methods： Four sheep were fed on either mixed hay （MH-diet） or MH-diet supplemented with 2% of Chinese herbal medicine （mixture of Astragalus root, Angelica root and Atractylodes rhizome; CHM-diet） over two 3S-day periods using a crossover design. The turnover rate of plasma glucose was measured with an isotope dilution method using [U-^13C]glucose. The rates of plasma leucine turnover and leucine oxidation, whole body protein synthesis （WBPS） and metabolic heat production were measured using the [1-^13C]leucine dilution and open circuit calorimetry. Results： Body weight gain of sheep was higher （P = 0.03） for CHM-diet than for MH-diet. Rumen pH was lower （P = 0.02）, concentration of rumen total volatile fatty acid tended to be higher （P = 0.05） and acetate was higher （P = 0.04） for CHM-diet than for MH-diet. Turnover rates of plasma glucose and leucine did not differ between diets. Oxidation rate of leucine tended to be higher （P = 0.06） for CHM-diet than for MH-diet, but the WBPS did not differ between diets. Metabolic heat production tended to be greater （P = 0.05） for CHM-diet than for MH-diet. Conclusions： The sheep fed on CHM-diet had a higher body weight gain and showed positive impacts on rumen fermentation and energy metabolism without resulting in any adverse response. Therefore, these results suggested that the Chinese herbal medicine mixture should be considered as a potential feed additive for sheep.展开更多
Four early lactating Holstein cows were used to study the effect of live yeast (LY, Actisaf®CNCM I-4407, Lesaffre Feed Additives, Marcq en Baroeul, France) supplementation on diet digestive utilization of dair...Four early lactating Holstein cows were used to study the effect of live yeast (LY, Actisaf®CNCM I-4407, Lesaffre Feed Additives, Marcq en Baroeul, France) supplementation on diet digestive utilization of dairy cows receiving concentrated corn silage-based diets with two rumen-degradable protein (RDP) levels. For a 33 d period, cows were fed a total mixed ration (TMR) containing an adequate level (AL) of RDP or a low level (LL, 30% below AL) by using soybean meal or tanned soybean meal, respectively: for 21 d with no LY addition followed by 12 d during which LY was added to the diet. The pH and redox potential (Eh) were recorded and ruminal fluid samples were collected over 3 consecutive days. Feces were collected individually over 48 h and individual dry matter intake (DMI) was measured for determining apparent nutrient digestibility. The effective degradability of individual feed ingredients composing both diets was evaluated with nylon bags technique. Structure of the ruminal bacterial community was studied and diversity index was calculated. Digestibility of organic matter (OM) and crude protein (CP) were lower for LL than those for AL. With LY, digestibility of OM and CP was increased: +2.4 and +0.8 points, for AL, and +3.7 and +5.9 points for LL, respectively. Live yeast reduced dietary N ruminal degradation with both AL and LL. Ruminal pH and Eh were lower with AL compared to LL: 5.95 and –167 mV vs. 6.13 and –144 mV. Live yeast increased ruminal total volatile fatty acids (VFA) (+8.6%), C2 (+10%), and C4 (+35%) contents for LL and decreased that of C3 (?9.8%) for AL. Neither the structure of bacterial populations of the rumen nor the diversity index (Shannon) was altered by treatments. Those results suggested a specific interest in using LY in RDP deficient diets for early lactating cows.展开更多
In complex digestion and metabolism of rumen, rumen microbes play a decisive role. Currently, the relationship between rumen microbial system and rumen metabolism has not been understood comprehensively, which will be...In complex digestion and metabolism of rumen, rumen microbes play a decisive role. Currently, the relationship between rumen microbial system and rumen metabolism has not been understood comprehensively, which will be reviewed in the paper, with particular focus on digestion and metabolism of bacteria, fungi and protozoa in the rumen of ruminants.展开更多
Background Dairy cows’lactation performance is the outcome of the crosstalk between ruminal microbial metabo-lism and host metabolism.However,it is still unclear to what extent the rumen microbiome and its metabolite...Background Dairy cows’lactation performance is the outcome of the crosstalk between ruminal microbial metabo-lism and host metabolism.However,it is still unclear to what extent the rumen microbiome and its metabolites,as well as the host metabolism,contribute to regulating the milk protein yield(MPY).Methods The rumen fluid,serum and milk of 12 Holstein cows with the same diet(45%coarseness ratio),parity(2–3 fetuses)and lactation days(120–150 d)were used for the microbiome and metabolome analysis.Rumen metabolism(rumen metabolome)and host metabolism(blood and milk metabolome)were connected using a weighted gene co-expression network(WGCNA)and the structural equation model(SEM)analyses.Results Two different ruminal enterotypes,with abundant Prevotella and Ruminococcus,were identified as type1 and type2.Of these,a higher MPY was found in cows with ruminal type2.Interestingly,[Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae(the differential bacteria)were the hub genera of the network.In addition,differential ruminal,serum and milk metabolome between enterotypes were identified,where the cows with type2 had higher L-tyrosine of rumen,ornithine and L-tryptophan of serum,and tetrahydroneopterin,palmitoyl-L-carnitine,S-lactoylglutathione of milk,which could provide more energy and substrate for MPY.Further,based on the identi-fied modules of ruminal microbiome,as well as ruminal serum and milk metabolome using WGCNA,the SEM analysis indicated that the key ruminal microbial module1,which contains the hub genera of the network([Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae)and high abundance of bacteria(Prevotella and Ruminococcus),could regulate the MPY by module7 of rumen,module2 of blood,and module7 of milk,which contained L-tyrosine and L-tryptophan.Therefore,in order to more clearly reveal the process of rumen bacterial regulation of MPY,we established the path of SEM based on the L-tyrosine,L-tryptophan and related components.The SEM based on the metabolites suggested that[Ruminococcus]gauvreauii group could inhibit the energy supply of serum tryptophan to MPY by milk S-lactoylglutathione,which could enhance pyruvate metabolism.Norank_f_Ruminococcaceae could increase the ruminal L-tyrosine,which could provide the substrate for MPY.Conclusion Our results indicated that the represented enterotype genera of Prevotella and Ruminococcus,and the hub genera of[Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae could regulate milk protein synthesis by affecting the ruminal L-tyrosine and L-tryptophan.Moreover,the combined analysis of enterotype,WGCNA and SEM could be used to connect rumen microbial metabolism with host metabolism,which provides a fundamental understanding of the crosstalk between host and microorganisms in regulating the synthesis of milk composition.展开更多
Slow-release non-protein nitrogen feed has a large market demand. It has a long research history, but its production technology needs further improvement in order to realize the industrial production of slow-release n...Slow-release non-protein nitrogen feed has a large market demand. It has a long research history, but its production technology needs further improvement in order to realize the industrial production of slow-release non-protein nitrogen extruded feed. By designing the best formula and using chelating and emulsifying process, the slow-release non-protein nitrogen extruded feed additives were produced. This product increases milk yield and improves milk quality, thus increasing economic efficiency.展开更多
基金Supported by New Diagnosis and Detection Technology Research for Major Animal Diseases in Cattle and Sheep(No.2016YFD0500901)
文摘[ Objective] The paper was to get effective soluble N protein to establish indirect ELISA method for Peste des petits ruminants （PPR）. [ Method] Soluble N protein with high expression was obtained from Escherichia coli expression system through codon optimization and optimization of expression conditions, and indirect ELISA detection method based on N protein was further established. [ Result] The assay had no cross reaction with other sheep pathogens. The intra- and inter-batch variation coefficients were less than 9%, indicating the method had good repeatability. Furthermore, totally 480 clinical serum samples were detec- ted by the assay, and the agreement rate with commercial ELISA kit （IDVET） was 98.33%. [ Conclusion] The study laid a foundation for further development of mature PPRV antibody detection kits.
基金Supported by National Key R&D Program for the Prevention and Control of Major Exotic Animal Diseases(2022YFD1800500)National Mutton Sheep Industrial Technology System(CARS39)+2 种基金Key Research and Development Program of Shandong Province(Major Science and Technology Innovation Project)(2021CXGC011306)Scientific Research Project of General Administration of Customs(2024HK033)Scientific Research Project of Jinan Customs(2023JK005).
文摘[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PPR.[Methods]Soluble N protein and NH fusion protein were successfully obtained in an Escherichia coli expression system by optimizing E.coli codon and expression conditions.Furthermore,based on purified soluble N protein and NH fusion protein,a double-antigen sandwich time-resolved fluorescence immunoassay method for detection of peste des petits ruminants virus(PPRV)was established.[Results]The method has high sensitivity and specificity and can specifically detect the antibody against PPRV in sheep serum,and it has no cross reaction with other related diseases.The method was used to detect 292 clinical samples,and compared with French IDVET competition ELISA kit.The coincidence rates of positive samples and negative samples from the two kinds of test kits were 92.47%and 97.26%,respectively,and the overall coincidence rate was 94.86%.The intra-group and inter-group coefficients of variation in the repeatability test were less than 10%.[Conclusions]Compared with the traditional ELISA method,the double-antigen sandwich time-resolved fluorescence immunoassay for detection of PPRV has equivalent sensitivity and specificity,and simple and rapid operation,and thus high application and popularization value.
文摘Background： The use of antibiotics in animal diets is facing negative feedback due to the hidden danger of drug residues to human health. Traditional Chinese herbal medicine has been used to replace antibiotics in the past two decades and played an increasingly important role in livestock production. The present study was carried out to assess the feeding effects of a traditional nourishing Chinese herbal medicine mixture on kinetics of plasma glucose, protein and energy metabolism in sheep. Ruminal fermentation characteristics were also determined. Methods： Four sheep were fed on either mixed hay （MH-diet） or MH-diet supplemented with 2% of Chinese herbal medicine （mixture of Astragalus root, Angelica root and Atractylodes rhizome; CHM-diet） over two 3S-day periods using a crossover design. The turnover rate of plasma glucose was measured with an isotope dilution method using [U-^13C]glucose. The rates of plasma leucine turnover and leucine oxidation, whole body protein synthesis （WBPS） and metabolic heat production were measured using the [1-^13C]leucine dilution and open circuit calorimetry. Results： Body weight gain of sheep was higher （P = 0.03） for CHM-diet than for MH-diet. Rumen pH was lower （P = 0.02）, concentration of rumen total volatile fatty acid tended to be higher （P = 0.05） and acetate was higher （P = 0.04） for CHM-diet than for MH-diet. Turnover rates of plasma glucose and leucine did not differ between diets. Oxidation rate of leucine tended to be higher （P = 0.06） for CHM-diet than for MH-diet, but the WBPS did not differ between diets. Metabolic heat production tended to be greater （P = 0.05） for CHM-diet than for MH-diet. Conclusions： The sheep fed on CHM-diet had a higher body weight gain and showed positive impacts on rumen fermentation and energy metabolism without resulting in any adverse response. Therefore, these results suggested that the Chinese herbal medicine mixture should be considered as a potential feed additive for sheep.
文摘Four early lactating Holstein cows were used to study the effect of live yeast (LY, Actisaf®CNCM I-4407, Lesaffre Feed Additives, Marcq en Baroeul, France) supplementation on diet digestive utilization of dairy cows receiving concentrated corn silage-based diets with two rumen-degradable protein (RDP) levels. For a 33 d period, cows were fed a total mixed ration (TMR) containing an adequate level (AL) of RDP or a low level (LL, 30% below AL) by using soybean meal or tanned soybean meal, respectively: for 21 d with no LY addition followed by 12 d during which LY was added to the diet. The pH and redox potential (Eh) were recorded and ruminal fluid samples were collected over 3 consecutive days. Feces were collected individually over 48 h and individual dry matter intake (DMI) was measured for determining apparent nutrient digestibility. The effective degradability of individual feed ingredients composing both diets was evaluated with nylon bags technique. Structure of the ruminal bacterial community was studied and diversity index was calculated. Digestibility of organic matter (OM) and crude protein (CP) were lower for LL than those for AL. With LY, digestibility of OM and CP was increased: +2.4 and +0.8 points, for AL, and +3.7 and +5.9 points for LL, respectively. Live yeast reduced dietary N ruminal degradation with both AL and LL. Ruminal pH and Eh were lower with AL compared to LL: 5.95 and –167 mV vs. 6.13 and –144 mV. Live yeast increased ruminal total volatile fatty acids (VFA) (+8.6%), C2 (+10%), and C4 (+35%) contents for LL and decreased that of C3 (?9.8%) for AL. Neither the structure of bacterial populations of the rumen nor the diversity index (Shannon) was altered by treatments. Those results suggested a specific interest in using LY in RDP deficient diets for early lactating cows.
基金Supported by National Natural Science Foundation of China(31040081)
文摘In complex digestion and metabolism of rumen, rumen microbes play a decisive role. Currently, the relationship between rumen microbial system and rumen metabolism has not been understood comprehensively, which will be reviewed in the paper, with particular focus on digestion and metabolism of bacteria, fungi and protozoa in the rumen of ruminants.
基金the National Natural Science Foundation of China(32272829,32072761,31902184)Shaanxi Provincial Science and Technology Association Young Talents Lifting Program Project(20220203).
文摘Background Dairy cows’lactation performance is the outcome of the crosstalk between ruminal microbial metabo-lism and host metabolism.However,it is still unclear to what extent the rumen microbiome and its metabolites,as well as the host metabolism,contribute to regulating the milk protein yield(MPY).Methods The rumen fluid,serum and milk of 12 Holstein cows with the same diet(45%coarseness ratio),parity(2–3 fetuses)and lactation days(120–150 d)were used for the microbiome and metabolome analysis.Rumen metabolism(rumen metabolome)and host metabolism(blood and milk metabolome)were connected using a weighted gene co-expression network(WGCNA)and the structural equation model(SEM)analyses.Results Two different ruminal enterotypes,with abundant Prevotella and Ruminococcus,were identified as type1 and type2.Of these,a higher MPY was found in cows with ruminal type2.Interestingly,[Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae(the differential bacteria)were the hub genera of the network.In addition,differential ruminal,serum and milk metabolome between enterotypes were identified,where the cows with type2 had higher L-tyrosine of rumen,ornithine and L-tryptophan of serum,and tetrahydroneopterin,palmitoyl-L-carnitine,S-lactoylglutathione of milk,which could provide more energy and substrate for MPY.Further,based on the identi-fied modules of ruminal microbiome,as well as ruminal serum and milk metabolome using WGCNA,the SEM analysis indicated that the key ruminal microbial module1,which contains the hub genera of the network([Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae)and high abundance of bacteria(Prevotella and Ruminococcus),could regulate the MPY by module7 of rumen,module2 of blood,and module7 of milk,which contained L-tyrosine and L-tryptophan.Therefore,in order to more clearly reveal the process of rumen bacterial regulation of MPY,we established the path of SEM based on the L-tyrosine,L-tryptophan and related components.The SEM based on the metabolites suggested that[Ruminococcus]gauvreauii group could inhibit the energy supply of serum tryptophan to MPY by milk S-lactoylglutathione,which could enhance pyruvate metabolism.Norank_f_Ruminococcaceae could increase the ruminal L-tyrosine,which could provide the substrate for MPY.Conclusion Our results indicated that the represented enterotype genera of Prevotella and Ruminococcus,and the hub genera of[Ruminococcus]gauvreauii group and norank_f_Ruminococcaceae could regulate milk protein synthesis by affecting the ruminal L-tyrosine and L-tryptophan.Moreover,the combined analysis of enterotype,WGCNA and SEM could be used to connect rumen microbial metabolism with host metabolism,which provides a fundamental understanding of the crosstalk between host and microorganisms in regulating the synthesis of milk composition.
基金funded by the Higher Vocational Colleges and Higher Junior Colleges Research Project of Heilongjiang Provincial Education Department (11515077)
文摘Slow-release non-protein nitrogen feed has a large market demand. It has a long research history, but its production technology needs further improvement in order to realize the industrial production of slow-release non-protein nitrogen extruded feed. By designing the best formula and using chelating and emulsifying process, the slow-release non-protein nitrogen extruded feed additives were produced. This product increases milk yield and improves milk quality, thus increasing economic efficiency.
