The performance of lactating dairy cows and composition of the fatty acids in milk fat were measured to investigate the effect of parity on conjugated linoleic acid(CLA)in milk fat and△~9-desaturase indices from Hols...The performance of lactating dairy cows and composition of the fatty acids in milk fat were measured to investigate the effect of parity on conjugated linoleic acid(CLA)in milk fat and△~9-desaturase indices from Holstein dairy cows.Milk samples of two hundred and thirty-five lactating Holstein dairy cows in north China were tested.In order to avoid possible confounding effects of diet and season,Holstein cows were fed a single diet and milk was sampled on the same day.The average content of cis-9,trans-11 CLA was 5.14 mg/g fatty acids in milk fat,varying between 1.5 and 10.5 mg/g fatty acids.The average content of cis-9,trans-11 CLA of primiparous cattle was 5.19 mg/g fatty acids in milk fat,while it was 5.11 mg/g fatty acids in multiparous animals,with no significant difference between them(P】0.05).There was a significant(P【0.05)difference between the△~9-desaturase index of cis-9 C14:l and cis-9 C16:l between primiparous and multiparous cattle but not(P】0.05)in the△~9-desaturase index of cis-9 C18:l and cis-9,trans-11 CLA.展开更多
To investigate the potential effects of Vaccaria segetalis, we employed the proteomic approach on the dairy cow mammary gland epithelial cells （DCMECs）. A total of 35 differentially expressed nuclear proteins were v...To investigate the potential effects of Vaccaria segetalis, we employed the proteomic approach on the dairy cow mammary gland epithelial cells （DCMECs）. A total of 35 differentially expressed nuclear proteins were visualized by 2-DE and silver nitrate staining. Of these, five proteins also displayed significant expression changes upon treatment of the water decoction from Vaccaria segetalis, and such alterations were further confirmed by RT-PCR. Together, at both the mRNA and protein levels, the water decoction from Vaccaria segetalis increased the expression of proteins ethylmalonic encephalopathy 1 （ETHE1）, vesicle amine transport protein 1 homolog （VAT1）, parkinson disease protein 7 homolog （Protein DJ-1）, proteasome subunit alpha type-2 （PSMA2） and SUMO-activating enzyme subunit 1 （SAE1）. This study would enable a better understanding of the molecular mechanisms underlying this water decoction effects at the protein level.展开更多
To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the reco...To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the recombinant plasmid pECFP-B by genetic engineering technique.Recombinant plasmid pECFP-B was transfected into dairy goat mammary gland epithelial to detect the bactericidal activity of CecropinB.The expression of CecropinB was also detected.The result of RT-PCR demonstrated CecropinB gene was expressed in transfected cells.CecropinB recombinant plasmid DNA was injected into udders and CecropinB was expressed in mammary gland,exhibiting bactericidal activity to Staphylococcus aureus in vivo experiments.展开更多
The major objective of this study was to determine the effect of corn straw or mixed diet on the small molecule metabolites of liver and milk production of healthy Chinese Holstein cows during lactation.In this study,...The major objective of this study was to determine the effect of corn straw or mixed diet on the small molecule metabolites of liver and milk production of healthy Chinese Holstein cows during lactation.In this study,metabolomic methods based on ultra performance liquid chromatography-mass spectrometry(LC-MS)were used to study the liver metabolites of dairy cows fed on corn straw diet or mixed diet.Ten healthy Chinese Holstein cows were randomly assigned to two groups,under the same management condition,fed different diets respectively,corn straw group(CS)or a mixture of alfalfa hay and Chinese wild rye hay mixed forage group(MF).All the cows were fed for 8 weeks and recorded body weight,dry matter intake,body condition score,fat,protein,lactose,milk yield and the total solids.Livers were sampled from each cow through a liver puncture needle for analysis of a significant difference in small molecule metabolites in cow liver samples from the two different diets.The results suggested that different diet types had significant effects on liver metabolism and milk components in dairy cows.The contents of milk fat,the total solids,milk protein,lactose,dry matter intake(DMI),milk yield,milk protein(%),lactose(%)and milk fat(%)of the corn straw group were significantly lower than those of the mixed forage group(p<0.05);the contents of phosphatidylcholine(PC),histidine,hypoxanthine and mridine in liver tissues of the corn straw group were significantly lower than those in the mixed forage group(p<0.05);acetylcarnitine,uric acid,triacylglycerol(TG),acetal phosphatidylcholine(plasmenyl-PC),acetalphosphatidylethanolamine(plasmenyl-PE)and sphingomyelin(SM)of the corn straw group were significantly higher than those in the mixed forage group(p<0.05).In summary,cows fed on mixed forage diet significantly improved milk yield and lactation performance clearly.展开更多
The main purpose of this study was to determine the effect of a corn straw or mixed diet on milk production, milk composition and the expression of genes associated with lactation in mid-lactation Chinese Holstein cow...The main purpose of this study was to determine the effect of a corn straw or mixed diet on milk production, milk composition and the expression of genes associated with lactation in mid-lactation Chinese Holstein cows. In this study, 10 healthy Chinese Holstein cows were randomly assigned to two groups and fed with different diets respectively, corn straw （CS） or mixed forage （MF） diet. CS group was fed roughage consisting of 53.8% corn straw only and the forge to concentrate （F ： C） ratio [dry matter （DM）] was about 40： 60. MF group was fed roughage consisting of 3.7% Chinese wildrye and 23.4% alfalfa hay, the forge to concentrate （F ： C） ratio （DM） was 70： 30. All the cows were fed 8 weeks and body weight, dry matter intake, body condition score, fat, protein, lactose, milk yield, total solid and somatic cell count （SCC） were recorded. Quantitative real-time PCR （qRT-PCR） was used to analyze cow mammary gland samples representing two different diets. The results suggested that different diet types had significant effects on milk yield, lactose, milk fat, milk protein, dry matter intake and somatic cell count in dairy cows, and cows fed MF diet improved milk production and lactation performance clearly （P〈0.05）. In addition, mRNA expression of genes ACC, roTOR, STATS, CSN2, PPAR）,, FABP3 and PTEN in MF group was extremely significantly higher than that in CS group （P〈0.05）. mRNA expression ofAKT1, FAS, SCD and SREBPlc in MF group was significantly higher than that in CS group （P〈0.01）. In summary, the milk yield and composition in mixed forage group were significantly improved than those in corn straw group.展开更多
Prolactin （PRL） is a versatile signaling molecule and regulates a variety of physiological processes, including mammary gland growth and differentiation and the synthesis of milk proteins. While PRL is known to be n...Prolactin （PRL） is a versatile signaling molecule and regulates a variety of physiological processes, including mammary gland growth and differentiation and the synthesis of milk proteins. While PRL is known to be necessary for high levels of milk protein expression, the mechanism by which the synthesis of milk proteins is stimulated at the transcript level is less known. A major modification in the transcript level is protein phosphorylation. To gain additional insights into the molecular mechanisms at the transcript level underlying PRL action on the dairy cow mammary epithelial cells （DCMECs）, nuclear phosphoproteins whose expression distinguishes proliferating regulated by PRL in DCMECs were identified. A phosphoprotein-enriched fraction from nuclear proteins was obtained by affinity chromatography, and a two-dimensional gel electrophoresis （2-DE） and matrix assisted laser desorption/ionization time of matrix-assisted laser desorption/ionization/time of flight mass spectrometry （MALDI-TOF MS） were used to identify the changes of nuclear phosphoproteins in DCMECs treated with prolactin. Seven proteins displaying~〉2-fold difference in abundance upon PRL treatment in DCMECs were identified by MALDI-TOF MS. The protein-GARS （GlyRS）, which belonged to the class-II aminoacyl-tRNA synthetase family, played a global role in the milk protein synthesis. SERPINH1 （Heat shock protein 47）, which was the first heat shock protein found to be a member of the serpin superfamily, regulated physiologic functions, such as complement activation, programmed cell death, and inflammatory processes. PRDX3, which belonged to a family of antioxidant enzymes, played an important role in scavenging intracellular reactive oxygen species （ROS）. ACTR1A, belonged to the actin family, which was associated with transport of p53 to the nucleus. Annexin A2, a Ca2＋-dependent phospholipid-binding protein, maintained the viability and cell cycle regulation of DCMECs. PSMB2 and PSMD10, which belonged to ubiquitin-proteasome system, were involved in several cellular processes, including cell cycle control, cellular stress response, intracellular signaling. This screening revealed that prolactin influenced the level of nuclear phosphoproteins in DCMECs. This result opens new avenues for the study of the molecular mechanism linked to the synthesis of milk proteins.展开更多
The lactating mammary gland is a prodigious protein-producing factory, but the milk protein synthesis mechanisms are not well understood. The major objective of this paper was to elucidate which genes and pathways wer...The lactating mammary gland is a prodigious protein-producing factory, but the milk protein synthesis mechanisms are not well understood. The major objective of this paper was to elucidate which genes and pathways were involved in the regulation of milk protein synthesis in the dairy goat mammary gland. Total 36 primiparous Guanzhong dairy goats were allotted in 12 groups according to their mammary development stages： days 90 and 150 of virgin, days 30, 90, and 150 of pregnancy, days 1, 10, 35, and 60 of lactation and days 3, 7, and 21 of involution （three animals per group）. Mammary tissue RNA was isolated for quantitative real- time RT-PCR of four casein genes alpha-s 1 casein （CSN 1S 1 ）, alpha-s2 casein （CSN 1 S2）, beta-casein （CSN2） and casein kappa （CSN3）, four whey protein genes lactoglobulin （LGB）, laetalbumin （LALBA）, laetofarrin （LTF）, and Whey acidic protein （WAP） and the genes which were potentially to regulate dairy goat milk protein synthesis at the level of transcription or translation [prolactin receptor （PRLR）, AKT1, signal transducers and activators of transcription 5 （STAT5）, E74-Like Factor 5 （ELF5）, eukaryotic translation initiation factor 4E binding protein 1 （EIF4E-BP1）, S6kinase （S6K） and caveolin 1]. The results showed that all genes were up-regulated in lactation period. The expressions of PRLR, AKT1, STAT5, ELF5, and S6K were similar to mRNA expressions of milk proteins. Our results indicated that milk protein synthesis in dairy goat mammary gland was possibly regulated by these genes.展开更多
Soybean meal （SBM） is commonly used for livestock feeds, but its application in diets for livestock is limited due to some antinutritional factors. The contents of methionine and lysine of soybean meal were promoted...Soybean meal （SBM） is commonly used for livestock feeds, but its application in diets for livestock is limited due to some antinutritional factors. The contents of methionine and lysine of soybean meal were promoted by Bacillus natto and Leuconostoc mesenteroides fermentation, benefial for the livestock feeds. It was crude protein （CP） 56.8%, methionine 43.56 mg · g^-1, and lysine 74.87 mg · g^-1, cows fed a diet with FSBM milk yield raised 14.2%, the change in the milk protein, the lactose and the dry matter content had also obvious increase. This convenient technique offers helpful exploration for industrialization of soybean meal fermentation.展开更多
According to the sequence of the bile salt hydrolase （BSH） gene of Bifidobacterium and the restriction enzyme cutting sites of expression vector pNZ8148, primers were designed and the bile salt hydrolase （BSH） gen...According to the sequence of the bile salt hydrolase （BSH） gene of Bifidobacterium and the restriction enzyme cutting sites of expression vector pNZ8148, primers were designed and the bile salt hydrolase （BSH） gene was gotten from Bacillus bifidus ATCC 29521 by PCR. BSH gene was inserted into lactic acid bacteria expression vector pNZ8148 to construct the recombinant pNZ8148-BSH. The recombinant pNZ8148-BSH was transferred into lactic acid bacteria NZ9000 with electrotransformation method. And the recombinant which could express BSH protein was obtained. It was identified by SDS-PAGE electrophoresis and activity verification. The result could provide a rationale reference for expressing BSH in lactic acid bacteria.展开更多
miRNA can regulate development and milk yield of the mammary gland through epigenetic mechanism, miRNA can directly and indirectly modulate the activity of the epigenetic machinery, target genes through post-inhibitio...miRNA can regulate development and milk yield of the mammary gland through epigenetic mechanism, miRNA can directly and indirectly modulate the activity of the epigenetic machinery, target genes through post-inhibition of translation initiation, mediate miRNA decay, target genes and inhibit the positive regulation, regulate tone modification, and regulate DNA methylation of target genes. Here we reviewed the role of miRNAs in mammary gland development and lactation. Researching miRNA in mammary gland development and lactation process, and understanding the response of the epigenetic mechanisms to external stimuli will be an important necessity to devise new technologies for maximizing their activity and milk production in the dairy cow.展开更多
The antagonistic activity of Lactobacillus acidophilus KLDS1.0901, KLDS1.0902, KLDSI.1003 and NCFM against Escherichia coli O157 ： H7 were investigated in this study. The culture supematants of all the L. acidophilus...The antagonistic activity of Lactobacillus acidophilus KLDS1.0901, KLDS1.0902, KLDSI.1003 and NCFM against Escherichia coli O157 ： H7 were investigated in this study. The culture supematants of all the L. acidophilus stains showed high bacteriostatic activities against E. coli O 157 ： H7 and the bacteriostatic substances of their Cell-Free Supernatants （CFS） were preliminarily determined from organic acids. The bacteriostatic activity from CFS or viable L. acidophilus against E. coli O157 ： H7 was also assessed by using co-incubation methods, CFS had high bactericidal activity against E. coli O157 ： H7, no viable E. coli O157 ： H7 was detected when 5×10^7 CfU ofE. coli O157 ： H7 was added to 5 mL of CFS and incubated at 37℃ for 2 h. However, L. acidophilus themselves had no bacteriostatic activity after directly contacted with E. coli O157 ： H7. The inhibition E. coli O157 ： H7 adhesion and colonization of L. acidophilus were also investigated based on competition, exclusion and displacement assays. L. acidophilus KLDS1.0901, KLDSI.1003 and NCFM strains were effective to displace E. coli O157 ： H7 from a Caco-2 cell layer in competition and exclusion assays. However, in displacement assay, all of the strains showed no significant antagonistic activities. Meanwhile, the probiotic potential of L. acidophilus strains was investigated based on adhesion assay to Caco-2 cells and anti- inflammatory effects by IL-8 produced in Caco-2 cells. The adhesion ability and anti-inflammatory effects of L. acidophilus strains showed a strain-dependent manner. In general, L. acidophilus KLDS 1.0901 and NCFM showed better probiotic potential than KLDS1.0902 and KLDSI.1003. Thus, the use ofL. acidophilus KLDS1.0901 and NCFM to prevent or treat of diseases associated induced E. coli O157 ： H7 in vivo was suggested.展开更多
Reversed-phase high-performance liquid chromatography （RP-HPLC） method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteris...Reversed-phase high-performance liquid chromatography （RP-HPLC） method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteristic of two columns were compared. Shiseido Proteonavi C4 column （250 mm×4.6 mm×5μm） and Shiseido CAPCELL PAK SG 300 C18 column （250 mm× 4.6 mm×5 μm） were used in the experiment. Phase A was composed of 0.1% （V/V） trifluoroacetic acid （TFA） in ultrapure water and Phase B （organic phase） was composed of 0.1% TFA in acetonitrile. Gradient elution was taken. Flow rate was 1 mL min-1. The detection wavelength was 215 nm. The injection volume was 20 μL and the column temperature was 30℃. The results showed that linear relationship was good and recovery of α-lactalbumin and β-lactoglobulin was 86.12%-104.38%, C18 column had stronger ability to resist acid and more stable, and the method with C4 column had excellent sensitivities and good separation.展开更多
A galactopoietic compound, identified as dibutyl phthalate（DBP）, was isolated from Vaccaria segetalis. The activity of DBP on lactation ability of dairy cow mammary gland epithelial cells（DCMECs） cultured in vitro...A galactopoietic compound, identified as dibutyl phthalate（DBP）, was isolated from Vaccaria segetalis. The activity of DBP on lactation ability of dairy cow mammary gland epithelial cells（DCMECs） cultured in vitro and dairy cow was evaluated. Results showed that DBP could promote cell viability, proliferation ability, lactose and β-casein secretion of DCMECs, which could also raise the milk yields of dairy cows significantly.展开更多
To further utilize bioactive substance such as bovine colostrum sIgA and IgG, sIgA and IgG were isolated and purified simultaneously by salting out, ultrafiltration and gel chromatography, etc. The analysis of results...To further utilize bioactive substance such as bovine colostrum sIgA and IgG, sIgA and IgG were isolated and purified simultaneously by salting out, ultrafiltration and gel chromatography, etc. The analysis of results were showed quantitatively by nonhydrogenized SDS-PAGE, and quanlities of sIgA and IgG were respectively detected by Western Blot. The results showed that the purity and yield of bovine colostrum sIgA were 85.3% and 42.8%, respectively, while the purity and yield of bovine colostrum IgG were respectively 97.2% and 64.4%. This preparative method provides theoretical and experimental foundation for sIgA industrial production.展开更多
With 3, 3’5, 5’-tetramethylbenzidine（TMB） as the detection substrate, a reliable and highly selective method was established and optimized for the determination of Lactoperoxidase（LP） activity in raw milk. The m...With 3, 3’5, 5’-tetramethylbenzidine（TMB） as the detection substrate, a reliable and highly selective method was established and optimized for the determination of Lactoperoxidase（LP） activity in raw milk. The method was based on the enzymatic reaction principle, where hydrogen peroxide oxidated TMB in the presence of LP. The optimized conditions of this assay system were obtained, consisting of 20 mmol · L-1 TMB solution, 0.6 mmol · L-1 hydrogen peroxide and 0.1 mol · L-1 Citric Acid（CA）/0.2 mol · L-1 disodium hydrogen phosphate（Na P） buffer（pH 4.8）. TMB detection method was applied to the analysis of LP in milk samples with a practical working concentration range from 2 to 14 mg · L-1. The intra-and inter-batch variation coefficients were all below 5%, indicating a good repeatability. Confirmation test between TMB method and 2, 2-azinobi（3-ethylbenzothiazoline-6-sulphonate） diammonium salt（ABTS） method was carried out, and the results of TMB assay were in accordance with that of ABTS method.展开更多
Feedstuff intake plays key role in impacting the yield and quality of milk. In this study, we discussed the contents of trace metal elements in cow milk impacted by different feedstuffs. We detected the contents of Pb...Feedstuff intake plays key role in impacting the yield and quality of milk. In this study, we discussed the contents of trace metal elements in cow milk impacted by different feedstuffs. We detected the contents of Pb, Cd, As, Cu, Mg, Ca, Fe and Zn in different feedstuffs and milk by atomic absorption spectrophotometry. The determinations of Pb, Cd and As contents were by graphite furnace A_AS and Cu, Mg, Ca, Fe and Zn was by flame atomic absorption spectrometry. The results showed that Pb, Cd, As and Cu presented in feedstuffs, but Pb, Cd and As were lowly detected in milk samples, and Cu was not detected in milk samples. The content of Mg in concentrates was lower than that in forages. However, the content of Mg in milk from concentrate feed was higher than that in forage feed. This suggested that the utilization of Mg in concentrate feed was higher than that in forage feed. The contents of Ca and Zn were opposite to Mg, and the utilization of Ca and Zn in forage feed was higher than that in concentrate feed. There was no obviously change law of Fe in different feedstuffs and milk samples. The relationship between the contents of trace metal elements in milk to feedstuffs would provide a theoretical basis for dairy farming. It would be useful for improving the milk yield and milk quality of dairy production.展开更多
Nutritional oil for infant formula food was microencapsulated by the spray drying method with coating materials including maltodextrin （MD）, soy protein isolate （SPI）, and emulsifier （soy lecithin）. Vegetable oi...Nutritional oil for infant formula food was microencapsulated by the spray drying method with coating materials including maltodextrin （MD）, soy protein isolate （SPI）, and emulsifier （soy lecithin）. Vegetable oil blend was prepared by mixing coconut oil, safflower oil and soybean oil at a ratio to achieve a fatty acid profile comparable to human milk fat （HMF）. The fatty acid composition of the product was determined by capillary gas chromatograph. As a result, the composition was as close as possible to that of HMF, it could be used for infant fomular food to make up some deficiencies of milk powder in nutrition and functional properties. Furthermore, the glass transition temperature （Tg） of the wall material was determined by DSC and its Tg was 66.42℃. It provided a theoretical basis for the storage of the product at the normal temperature.展开更多
In this study, the Single Factor Test（SFT） was used to optimize the pre-freezing conditions of L. plantarum KLDS1.0391（KLDS1.0391）. Further, the Freeze-Drying Protective Agents（FDPA） of KLDS1.0391 was optimized ...In this study, the Single Factor Test（SFT） was used to optimize the pre-freezing conditions of L. plantarum KLDS1.0391（KLDS1.0391）. Further, the Freeze-Drying Protective Agents（FDPA） of KLDS1.0391 was optimized by Response Surface Methodology（RSM）. The optimum pretreatment conditions were as the follows： initial concentration of KLDS1.0391 was 1011 CFU · m L-1 and KLDS1.0391 was pre-freezed at –80℃ for 8 h to achieve the survival rate of 46.21%. The main components of FDPA were skim milk, sucrose, sodium glutamate and Tween-80. And the influence of four factors on the survival rate of KLDS1.0391 in freeze-drying was in order as the follows： skim milk〉sucrose〉Tween-80〉sodium glutamate. The optimal FDPA composition was skim milk 11.3%, sucrose 9.8%, sodium glutamate 5.1% and Tween-80 0.2%. Under the above conditions, the survival rate of the cells was 82.98%. Comparing the predicted values, the relative error was 0.37% and the difference was not significant, which indicated that the established model could effectively reflect the actual protection of FDPA to KLDS1.0391.展开更多
sIgA in bovine colostrum was purified by ultrafiltration and enzymolysis methods in this experiment, and the prepared substance was detected by Western Blot. The purity and yield were up to 73.6% and 65.2%, respective...sIgA in bovine colostrum was purified by ultrafiltration and enzymolysis methods in this experiment, and the prepared substance was detected by Western Blot. The purity and yield were up to 73.6% and 65.2%, respectively. This convenient technique offers helpful exploration for production of bovine colostrum sIgA展开更多
The objective of this work was to study the influence of cooking time and cooling rate on functionality and microstructure of processed cheese spreads. When the cooking time was 20 min, the hardness and apparent visco...The objective of this work was to study the influence of cooking time and cooling rate on functionality and microstructure of processed cheese spreads. When the cooking time was 20 min, the hardness and apparent viscosity were increased, and formed a homogenous, dense, and three-dimensional protein network, and a stronger gel was formed at this time. The slow cooling can increase the hardness and apparent viscosity of products, protein wall was thicker, and network was closer, so products can formed a stronger gel structure. The influence of cooking time on the functional properties was more significant than the influence of rapid cooling.展开更多
文摘The performance of lactating dairy cows and composition of the fatty acids in milk fat were measured to investigate the effect of parity on conjugated linoleic acid(CLA)in milk fat and△~9-desaturase indices from Holstein dairy cows.Milk samples of two hundred and thirty-five lactating Holstein dairy cows in north China were tested.In order to avoid possible confounding effects of diet and season,Holstein cows were fed a single diet and milk was sampled on the same day.The average content of cis-9,trans-11 CLA was 5.14 mg/g fatty acids in milk fat,varying between 1.5 and 10.5 mg/g fatty acids.The average content of cis-9,trans-11 CLA of primiparous cattle was 5.19 mg/g fatty acids in milk fat,while it was 5.11 mg/g fatty acids in multiparous animals,with no significant difference between them(P】0.05).There was a significant(P【0.05)difference between the△~9-desaturase index of cis-9 C14:l and cis-9 C16:l between primiparous and multiparous cattle but not(P】0.05)in the△~9-desaturase index of cis-9 C18:l and cis-9,trans-11 CLA.
基金Supported by Major State Basic Research Development Program of China (973 Program, 2011CB100804)
文摘To investigate the potential effects of Vaccaria segetalis, we employed the proteomic approach on the dairy cow mammary gland epithelial cells （DCMECs）. A total of 35 differentially expressed nuclear proteins were visualized by 2-DE and silver nitrate staining. Of these, five proteins also displayed significant expression changes upon treatment of the water decoction from Vaccaria segetalis, and such alterations were further confirmed by RT-PCR. Together, at both the mRNA and protein levels, the water decoction from Vaccaria segetalis increased the expression of proteins ethylmalonic encephalopathy 1 （ETHE1）, vesicle amine transport protein 1 homolog （VAT1）, parkinson disease protein 7 homolog （Protein DJ-1）, proteasome subunit alpha type-2 （PSMA2） and SUMO-activating enzyme subunit 1 （SAE1）. This study would enable a better understanding of the molecular mechanisms underlying this water decoction effects at the protein level.
基金Supported by Key Teachers Foundation of Education Office of Heilongjiang Province,2005(1055G005)
文摘To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the recombinant plasmid pECFP-B by genetic engineering technique.Recombinant plasmid pECFP-B was transfected into dairy goat mammary gland epithelial to detect the bactericidal activity of CecropinB.The expression of CecropinB was also detected.The result of RT-PCR demonstrated CecropinB gene was expressed in transfected cells.CecropinB recombinant plasmid DNA was injected into udders and CecropinB was expressed in mammary gland,exhibiting bactericidal activity to Staphylococcus aureus in vivo experiments.
基金Supported by the National Natural Science Foundation of China(31101784)Funds for Young Researchers from Northeast Agricultural University(14QC43)。
文摘The major objective of this study was to determine the effect of corn straw or mixed diet on the small molecule metabolites of liver and milk production of healthy Chinese Holstein cows during lactation.In this study,metabolomic methods based on ultra performance liquid chromatography-mass spectrometry(LC-MS)were used to study the liver metabolites of dairy cows fed on corn straw diet or mixed diet.Ten healthy Chinese Holstein cows were randomly assigned to two groups,under the same management condition,fed different diets respectively,corn straw group(CS)or a mixture of alfalfa hay and Chinese wild rye hay mixed forage group(MF).All the cows were fed for 8 weeks and recorded body weight,dry matter intake,body condition score,fat,protein,lactose,milk yield and the total solids.Livers were sampled from each cow through a liver puncture needle for analysis of a significant difference in small molecule metabolites in cow liver samples from the two different diets.The results suggested that different diet types had significant effects on liver metabolism and milk components in dairy cows.The contents of milk fat,the total solids,milk protein,lactose,dry matter intake(DMI),milk yield,milk protein(%),lactose(%)and milk fat(%)of the corn straw group were significantly lower than those of the mixed forage group(p<0.05);the contents of phosphatidylcholine(PC),histidine,hypoxanthine and mridine in liver tissues of the corn straw group were significantly lower than those in the mixed forage group(p<0.05);acetylcarnitine,uric acid,triacylglycerol(TG),acetal phosphatidylcholine(plasmenyl-PC),acetalphosphatidylethanolamine(plasmenyl-PE)and sphingomyelin(SM)of the corn straw group were significantly higher than those in the mixed forage group(p<0.05).In summary,cows fed on mixed forage diet significantly improved milk yield and lactation performance clearly.
基金Supported by Fund of the National Basic Research Program of China(973)(2011CB100804)
文摘The main purpose of this study was to determine the effect of a corn straw or mixed diet on milk production, milk composition and the expression of genes associated with lactation in mid-lactation Chinese Holstein cows. In this study, 10 healthy Chinese Holstein cows were randomly assigned to two groups and fed with different diets respectively, corn straw （CS） or mixed forage （MF） diet. CS group was fed roughage consisting of 53.8% corn straw only and the forge to concentrate （F ： C） ratio [dry matter （DM）] was about 40： 60. MF group was fed roughage consisting of 3.7% Chinese wildrye and 23.4% alfalfa hay, the forge to concentrate （F ： C） ratio （DM） was 70： 30. All the cows were fed 8 weeks and body weight, dry matter intake, body condition score, fat, protein, lactose, milk yield, total solid and somatic cell count （SCC） were recorded. Quantitative real-time PCR （qRT-PCR） was used to analyze cow mammary gland samples representing two different diets. The results suggested that different diet types had significant effects on milk yield, lactose, milk fat, milk protein, dry matter intake and somatic cell count in dairy cows, and cows fed MF diet improved milk production and lactation performance clearly （P〈0.05）. In addition, mRNA expression of genes ACC, roTOR, STATS, CSN2, PPAR）,, FABP3 and PTEN in MF group was extremely significantly higher than that in CS group （P〈0.05）. mRNA expression ofAKT1, FAS, SCD and SREBPlc in MF group was significantly higher than that in CS group （P〈0.01）. In summary, the milk yield and composition in mixed forage group were significantly improved than those in corn straw group.
基金Supported by Major State Basic Research Development Program of China(973 Program,2011CB100804)
文摘Prolactin （PRL） is a versatile signaling molecule and regulates a variety of physiological processes, including mammary gland growth and differentiation and the synthesis of milk proteins. While PRL is known to be necessary for high levels of milk protein expression, the mechanism by which the synthesis of milk proteins is stimulated at the transcript level is less known. A major modification in the transcript level is protein phosphorylation. To gain additional insights into the molecular mechanisms at the transcript level underlying PRL action on the dairy cow mammary epithelial cells （DCMECs）, nuclear phosphoproteins whose expression distinguishes proliferating regulated by PRL in DCMECs were identified. A phosphoprotein-enriched fraction from nuclear proteins was obtained by affinity chromatography, and a two-dimensional gel electrophoresis （2-DE） and matrix assisted laser desorption/ionization time of matrix-assisted laser desorption/ionization/time of flight mass spectrometry （MALDI-TOF MS） were used to identify the changes of nuclear phosphoproteins in DCMECs treated with prolactin. Seven proteins displaying~〉2-fold difference in abundance upon PRL treatment in DCMECs were identified by MALDI-TOF MS. The protein-GARS （GlyRS）, which belonged to the class-II aminoacyl-tRNA synthetase family, played a global role in the milk protein synthesis. SERPINH1 （Heat shock protein 47）, which was the first heat shock protein found to be a member of the serpin superfamily, regulated physiologic functions, such as complement activation, programmed cell death, and inflammatory processes. PRDX3, which belonged to a family of antioxidant enzymes, played an important role in scavenging intracellular reactive oxygen species （ROS）. ACTR1A, belonged to the actin family, which was associated with transport of p53 to the nucleus. Annexin A2, a Ca2＋-dependent phospholipid-binding protein, maintained the viability and cell cycle regulation of DCMECs. PSMB2 and PSMD10, which belonged to ubiquitin-proteasome system, were involved in several cellular processes, including cell cycle control, cellular stress response, intracellular signaling. This screening revealed that prolactin influenced the level of nuclear phosphoproteins in DCMECs. This result opens new avenues for the study of the molecular mechanism linked to the synthesis of milk proteins.
基金Supported by the National Basic Research Program of China(973 Program,2011CB100804)the National Natural Science Foundation of China(31101784)Funds for Young Researchers from Northeast Agricultural University(14QC43)
文摘The lactating mammary gland is a prodigious protein-producing factory, but the milk protein synthesis mechanisms are not well understood. The major objective of this paper was to elucidate which genes and pathways were involved in the regulation of milk protein synthesis in the dairy goat mammary gland. Total 36 primiparous Guanzhong dairy goats were allotted in 12 groups according to their mammary development stages： days 90 and 150 of virgin, days 30, 90, and 150 of pregnancy, days 1, 10, 35, and 60 of lactation and days 3, 7, and 21 of involution （three animals per group）. Mammary tissue RNA was isolated for quantitative real- time RT-PCR of four casein genes alpha-s 1 casein （CSN 1S 1 ）, alpha-s2 casein （CSN 1 S2）, beta-casein （CSN2） and casein kappa （CSN3）, four whey protein genes lactoglobulin （LGB）, laetalbumin （LALBA）, laetofarrin （LTF）, and Whey acidic protein （WAP） and the genes which were potentially to regulate dairy goat milk protein synthesis at the level of transcription or translation [prolactin receptor （PRLR）, AKT1, signal transducers and activators of transcription 5 （STAT5）, E74-Like Factor 5 （ELF5）, eukaryotic translation initiation factor 4E binding protein 1 （EIF4E-BP1）, S6kinase （S6K） and caveolin 1]. The results showed that all genes were up-regulated in lactation period. The expressions of PRLR, AKT1, STAT5, ELF5, and S6K were similar to mRNA expressions of milk proteins. Our results indicated that milk protein synthesis in dairy goat mammary gland was possibly regulated by these genes.
基金Supported by Science and Technology Foundation (GB08B401-02)Science and Technology for Youth in Heilongjiang Province (QC07C35)
文摘Soybean meal （SBM） is commonly used for livestock feeds, but its application in diets for livestock is limited due to some antinutritional factors. The contents of methionine and lysine of soybean meal were promoted by Bacillus natto and Leuconostoc mesenteroides fermentation, benefial for the livestock feeds. It was crude protein （CP） 56.8%, methionine 43.56 mg · g^-1, and lysine 74.87 mg · g^-1, cows fed a diet with FSBM milk yield raised 14.2%, the change in the milk protein, the lactose and the dry matter content had also obvious increase. This convenient technique offers helpful exploration for industrialization of soybean meal fermentation.
基金Supported by 863 Projects (2008AA10Z311)National Science and Technology Support Projects (2009BADB9B06)+1 种基金Started Post-doctoral Research Grant of Heilongjiang Province (LBH-Q07023)Harbin Technological Innovation of Special Funds (2007RFQXN020)
文摘According to the sequence of the bile salt hydrolase （BSH） gene of Bifidobacterium and the restriction enzyme cutting sites of expression vector pNZ8148, primers were designed and the bile salt hydrolase （BSH） gene was gotten from Bacillus bifidus ATCC 29521 by PCR. BSH gene was inserted into lactic acid bacteria expression vector pNZ8148 to construct the recombinant pNZ8148-BSH. The recombinant pNZ8148-BSH was transferred into lactic acid bacteria NZ9000 with electrotransformation method. And the recombinant which could express BSH protein was obtained. It was identified by SDS-PAGE electrophoresis and activity verification. The result could provide a rationale reference for expressing BSH in lactic acid bacteria.
基金Support by the Natural Science Foundation of China(31072103)
文摘miRNA can regulate development and milk yield of the mammary gland through epigenetic mechanism, miRNA can directly and indirectly modulate the activity of the epigenetic machinery, target genes through post-inhibition of translation initiation, mediate miRNA decay, target genes and inhibit the positive regulation, regulate tone modification, and regulate DNA methylation of target genes. Here we reviewed the role of miRNAs in mammary gland development and lactation. Researching miRNA in mammary gland development and lactation process, and understanding the response of the epigenetic mechanisms to external stimuli will be an important necessity to devise new technologies for maximizing their activity and milk production in the dairy cow.
基金Supported by the Agro-scientific Research of China(201203009)the Ministry of Agriculture,China
文摘The antagonistic activity of Lactobacillus acidophilus KLDS1.0901, KLDS1.0902, KLDSI.1003 and NCFM against Escherichia coli O157 ： H7 were investigated in this study. The culture supematants of all the L. acidophilus stains showed high bacteriostatic activities against E. coli O 157 ： H7 and the bacteriostatic substances of their Cell-Free Supernatants （CFS） were preliminarily determined from organic acids. The bacteriostatic activity from CFS or viable L. acidophilus against E. coli O157 ： H7 was also assessed by using co-incubation methods, CFS had high bactericidal activity against E. coli O157 ： H7, no viable E. coli O157 ： H7 was detected when 5×10^7 CfU ofE. coli O157 ： H7 was added to 5 mL of CFS and incubated at 37℃ for 2 h. However, L. acidophilus themselves had no bacteriostatic activity after directly contacted with E. coli O157 ： H7. The inhibition E. coli O157 ： H7 adhesion and colonization of L. acidophilus were also investigated based on competition, exclusion and displacement assays. L. acidophilus KLDS1.0901, KLDSI.1003 and NCFM strains were effective to displace E. coli O157 ： H7 from a Caco-2 cell layer in competition and exclusion assays. However, in displacement assay, all of the strains showed no significant antagonistic activities. Meanwhile, the probiotic potential of L. acidophilus strains was investigated based on adhesion assay to Caco-2 cells and anti- inflammatory effects by IL-8 produced in Caco-2 cells. The adhesion ability and anti-inflammatory effects of L. acidophilus strains showed a strain-dependent manner. In general, L. acidophilus KLDS 1.0901 and NCFM showed better probiotic potential than KLDS1.0902 and KLDSI.1003. Thus, the use ofL. acidophilus KLDS1.0901 and NCFM to prevent or treat of diseases associated induced E. coli O157 ： H7 in vivo was suggested.
基金Supported by the Project of Science & Technology Plans in Heilongjiang Province in the 11th Five-year Period (GB07B407)
文摘Reversed-phase high-performance liquid chromatography （RP-HPLC） method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteristic of two columns were compared. Shiseido Proteonavi C4 column （250 mm×4.6 mm×5μm） and Shiseido CAPCELL PAK SG 300 C18 column （250 mm× 4.6 mm×5 μm） were used in the experiment. Phase A was composed of 0.1% （V/V） trifluoroacetic acid （TFA） in ultrapure water and Phase B （organic phase） was composed of 0.1% TFA in acetonitrile. Gradient elution was taken. Flow rate was 1 mL min-1. The detection wavelength was 215 nm. The injection volume was 20 μL and the column temperature was 30℃. The results showed that linear relationship was good and recovery of α-lactalbumin and β-lactoglobulin was 86.12%-104.38%, C18 column had stronger ability to resist acid and more stable, and the method with C4 column had excellent sensitivities and good separation.
基金Supported by "863" Project of Ministry of Science and Technology of China(2013AA102504-03)Talents Foundation of Northeast Agricultural University(2010RCB47,2010RCB55)
文摘A galactopoietic compound, identified as dibutyl phthalate（DBP）, was isolated from Vaccaria segetalis. The activity of DBP on lactation ability of dairy cow mammary gland epithelial cells（DCMECs） cultured in vitro and dairy cow was evaluated. Results showed that DBP could promote cell viability, proliferation ability, lactose and β-casein secretion of DCMECs, which could also raise the milk yields of dairy cows significantly.
文摘To further utilize bioactive substance such as bovine colostrum sIgA and IgG, sIgA and IgG were isolated and purified simultaneously by salting out, ultrafiltration and gel chromatography, etc. The analysis of results were showed quantitatively by nonhydrogenized SDS-PAGE, and quanlities of sIgA and IgG were respectively detected by Western Blot. The results showed that the purity and yield of bovine colostrum sIgA were 85.3% and 42.8%, respectively, while the purity and yield of bovine colostrum IgG were respectively 97.2% and 64.4%. This preparative method provides theoretical and experimental foundation for sIgA industrial production.
基金Supported by Project for Research and Development of Harbin Aapplication Technology(2016RAQXJ046)the National "Twelfth Five-year" Plan for Science and Technology Support Program of China(2013BAD18B06)
文摘With 3, 3’5, 5’-tetramethylbenzidine（TMB） as the detection substrate, a reliable and highly selective method was established and optimized for the determination of Lactoperoxidase（LP） activity in raw milk. The method was based on the enzymatic reaction principle, where hydrogen peroxide oxidated TMB in the presence of LP. The optimized conditions of this assay system were obtained, consisting of 20 mmol · L-1 TMB solution, 0.6 mmol · L-1 hydrogen peroxide and 0.1 mol · L-1 Citric Acid（CA）/0.2 mol · L-1 disodium hydrogen phosphate（Na P） buffer（pH 4.8）. TMB detection method was applied to the analysis of LP in milk samples with a practical working concentration range from 2 to 14 mg · L-1. The intra-and inter-batch variation coefficients were all below 5%, indicating a good repeatability. Confirmation test between TMB method and 2, 2-azinobi（3-ethylbenzothiazoline-6-sulphonate） diammonium salt（ABTS） method was carried out, and the results of TMB assay were in accordance with that of ABTS method.
文摘Feedstuff intake plays key role in impacting the yield and quality of milk. In this study, we discussed the contents of trace metal elements in cow milk impacted by different feedstuffs. We detected the contents of Pb, Cd, As, Cu, Mg, Ca, Fe and Zn in different feedstuffs and milk by atomic absorption spectrophotometry. The determinations of Pb, Cd and As contents were by graphite furnace A_AS and Cu, Mg, Ca, Fe and Zn was by flame atomic absorption spectrometry. The results showed that Pb, Cd, As and Cu presented in feedstuffs, but Pb, Cd and As were lowly detected in milk samples, and Cu was not detected in milk samples. The content of Mg in concentrates was lower than that in forages. However, the content of Mg in milk from concentrate feed was higher than that in forage feed. This suggested that the utilization of Mg in concentrate feed was higher than that in forage feed. The contents of Ca and Zn were opposite to Mg, and the utilization of Ca and Zn in forage feed was higher than that in concentrate feed. There was no obviously change law of Fe in different feedstuffs and milk samples. The relationship between the contents of trace metal elements in milk to feedstuffs would provide a theoretical basis for dairy farming. It would be useful for improving the milk yield and milk quality of dairy production.
文摘Nutritional oil for infant formula food was microencapsulated by the spray drying method with coating materials including maltodextrin （MD）, soy protein isolate （SPI）, and emulsifier （soy lecithin）. Vegetable oil blend was prepared by mixing coconut oil, safflower oil and soybean oil at a ratio to achieve a fatty acid profile comparable to human milk fat （HMF）. The fatty acid composition of the product was determined by capillary gas chromatograph. As a result, the composition was as close as possible to that of HMF, it could be used for infant fomular food to make up some deficiencies of milk powder in nutrition and functional properties. Furthermore, the glass transition temperature （Tg） of the wall material was determined by DSC and its Tg was 66.42℃. It provided a theoretical basis for the storage of the product at the normal temperature.
基金Supported by the Outstanding Youth Scientists Foundation of Harbin City(2014RFYXJ006)
文摘In this study, the Single Factor Test（SFT） was used to optimize the pre-freezing conditions of L. plantarum KLDS1.0391（KLDS1.0391）. Further, the Freeze-Drying Protective Agents（FDPA） of KLDS1.0391 was optimized by Response Surface Methodology（RSM）. The optimum pretreatment conditions were as the follows： initial concentration of KLDS1.0391 was 1011 CFU · m L-1 and KLDS1.0391 was pre-freezed at –80℃ for 8 h to achieve the survival rate of 46.21%. The main components of FDPA were skim milk, sucrose, sodium glutamate and Tween-80. And the influence of four factors on the survival rate of KLDS1.0391 in freeze-drying was in order as the follows： skim milk〉sucrose〉Tween-80〉sodium glutamate. The optimal FDPA composition was skim milk 11.3%, sucrose 9.8%, sodium glutamate 5.1% and Tween-80 0.2%. Under the above conditions, the survival rate of the cells was 82.98%. Comparing the predicted values, the relative error was 0.37% and the difference was not significant, which indicated that the established model could effectively reflect the actual protection of FDPA to KLDS1.0391.
基金the Science and Technology Foundation of Heilongjiang Province (GA06B201-4)
文摘sIgA in bovine colostrum was purified by ultrafiltration and enzymolysis methods in this experiment, and the prepared substance was detected by Western Blot. The purity and yield were up to 73.6% and 65.2%, respectively. This convenient technique offers helpful exploration for production of bovine colostrum sIgA
基金Supported by Tackle Problem Project of Science and Technology Department of Heilongjiang Province (GC05B404)
文摘The objective of this work was to study the influence of cooking time and cooling rate on functionality and microstructure of processed cheese spreads. When the cooking time was 20 min, the hardness and apparent viscosity were increased, and formed a homogenous, dense, and three-dimensional protein network, and a stronger gel was formed at this time. The slow cooling can increase the hardness and apparent viscosity of products, protein wall was thicker, and network was closer, so products can formed a stronger gel structure. The influence of cooking time on the functional properties was more significant than the influence of rapid cooling.
