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The Regulation of pH Value of Liquid Feed on Blood Gas Parameters in Holstein Bull Calves TU Yan;QIU Guo-liang; ZHOU Yi; YUN Qiang; QI Dong; WANG Jia-jie; DIAO Qi-yu Scientia Agricultura Sinica    2014, 47 (17): 3465-3474.   DOI: 10.3864/j.issn.0578-1752.2014.17.014 Abstract （347）   HTML （3）    PDF （643KB）（749）       Knowledge map    Save 【Objective】 Due to the immature gastrointestinal function and low ability digest dietary, diarrhea or other nutritional diseases were easy to arise in suckling calves that bring about low survival rate. Compared with adult cattle, liquid milk replacer is the main feed for the early weaned calves. Thus, acidity of milk replacer emulsions has more important effect on calf health. But the change regulation, the normal value scope of blood gas parameters in calves are not in any systematic manner, so that it is unable to determine whether the appropriate dietary acidity is. To study the change regulations of blood gas parameter in calves, an experiment was carried out to investigate the variations of blood gas parameters in calves which fed liquid feed with different pH. 【Method】The pH values of a milk replacer (6.2, 5.5, 5.0 or 4.5) and the ratio of vegetable protein to total protein in the milk replacer (50% or 80%) were used to form an 2 × 4 factorial design in this experiment. Forty-eight neonatal healthy Chinese Holstein male calves were allotted into eight groups and each group was fed with one of the 8 milk replacer emulsions. The experiment lasted 63 d with 21 d for adaptation and 42 d for test. Growth performance was determined fortnightly, the intake of milk replacers and pellet diet were recorded every day. Blood samples were collected on 21 d and 49 d. The pH value, partial pressure of carbon dioxide (pCO2), partial pressure of oxygen (pO2), oxygen saturation (SO2), the concentration of [HCO3-](HCO3-), actual base excess (ABE), total carbon dioxide volume (TCO2), standard bicarbonate concentration (SBC), standard base excess (SBE) of the blood samples were determined. Data of growth performance or blood gas parameters were analyzed using the GLM or MIXED procedure of SAS software, the regression equations between blood gas parameters and milk replacer emulsion pH value were established by REG procedure, and the changes of blood gas parameters were analyzed using normal distribution test. 【Result】The results showed that the average daily gain were improved while the pH value of milk replacer decreased (P＜0.05). The pH value of milk replacer siginificantly affected blood pO2, SO2, HCO3-, ABE, TCO2, SBC and SBE (P＜0.01). pH value, pCO2, pO2, SO2, HCO3- and TCO2 showed a quadratic curve (P＜0.05) as milk replacer emulsion pH value decreased with R2 as 0.9201, 0.8481, 0.8600, 0.9445, 0.8631, 0.8141, respectively, meanwhile ABE, SBC, SBE were showed a linear turn (P＜0.05) with R2 as 0.9060, 0.8126, 0.8298. Blood pH and pCO2 were varied remarkably with the age of day increasing (P＜0.05). Blood gas parameters were in line with the normal distribution (P＞0.05) except for HCO3-, ABE, SBE. While the ratio of vegetable protein to total protein of milk replacer increased from 50% to 80%, the average daily gain and dry matter intake of calves decreased, but blood gas parameters were unaffected (P＞0.05). 【Conclusion】The blood HCO3-, ABE, TCO2, SBC,and SBE of calves aged 21 to 63 days significantly decreased with the pH value of milk replacer dropping. Blood gas parameters can be used as sensitive indices to evaluate the effect of dietary acidity on calves. Reference | Related Articles | Metrics

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Differences in Expression of HSPA2 in Different Tissues and Organs of Yak WEN Ze-xing, YU Si-jiu, ZHAI Yu-jia, YANG Kun, LIU Peng-gang, HE Jun-feng, CUI Yan Scientia Agricultura Sinica    2014, 47 (17): 3475-3482.   DOI: 10.3864/j.issn.0578-1752.2014.17.015 Abstract （431）   HTML （3）    PDF （878KB）（870）       Knowledge map    Save 【Objective】 This experiment was conducted to study the differences in expression of heat shock protein 70-2 (HSPA2) in different tissues and organs of yak. 【Method】 Three 1-year-old Qinghai Plateau male yaks were used in the present study. Under normal physiological conditions, the tissues and organs were obtained from healthy yak in mid-september 2013 (heart, liver, spleen, lung, kidney, brain and testis). The RNA were extracted from different tissues and organs of yak, and the RNA were reverse-transcribed into first-strand cDNA. The primers were designed specifically according to the Bos grunniens HSPA2 and β-actin gene sequences (HSPA2:KC790105.1, β-actin: DQ838049.1). Firstly, RT-PCR were used to verify the real-time quantitative PCR (RT-qPCR) could be applied to the determination of the differences in expression of HSPA2 gene in different tissues and organs of yak. Then, real time quantitative PCR was applied to analyze the expression of HSPA2 gene. Paraformaldehyde was used to fix different tissues and organs of yak, and then made into paraffin sections. The protein localization of HSPA2 was measured by immunohistochemistry. The Image-Pro Plus 6.0 software was used to analyze immunohistochemistry, the integrated optical density of HSPA2 positive reaction was measured, and absorbance was analyzed. The SPSS19.0 software was used for significance of difference analysis. 【Result】 RT-PCR results showed that RT-qPCR can be applied to the determination of differences in expression of HSPA2 gene in different tissues and organs of yak. The results of RT-qPCR showed that comparative expression quantity of gene HSPA2 in testis was 83.33 fold than that in brain, 97.09 fold in kidney, 111.11 fold in heart,133.33 fold in spleen, 222.22 fold in lung and 285.71 fold in liver, respectively. Immunohistochemistry showed that HSPA2 expressed in the testis, kidney, brain, heart, lung, liver and spleen of yak. HSPA2 positive reaction was observed in yak renal cortical tubular, medullary tubules, cortical hippocampal CA1 region, cerebellar cortex, cardiac cells, alveolar epithelial cells, liver cells, spleen marginal zone and red pulp. The most positive reaction was found in the cytoplasm, and positive reaction was rarely found in the nucleus. In the testis seminiferous tubules, HSPA2 positive reaction was showed in both the cytoplasm and nucleus of spermatogenic cells. No positive reaction was observed in the negative control group. Analysis of integrated optical density values showed that the HSPA2 protein expression in testis was the strongest, and followed by the brain, kidney, heart, lung and liver, and its expression in spleen was the weakest. 【Conclusion】 By studying the gene and protein levels, the findings of the study showed the differences in expression of HSPA2 in different tissues and organs of yak. The expression of HSPA2 gene and protein in testis were higher than that in brain, kidney, heart, spleen, lung and liver, which suggests that HSPA2 may be associated with testicular reproductive function. Reference | Related Articles | Metrics Cited: Baidu(1)

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Transcription Profiles of Immune-Related Genes in Chickens Infected by Salmonella Enteritidis and Poly(I:C) SUN Yan, LI Jian-chao, LI Peng, ZHENG Mai-qing, LIU Ran-ran, LI Qing-he, WEN Jie, ZHAO Gui-ping Scientia Agricultura Sinica    2014, 47 (17): 3483-3491.   DOI: 10.3864/j.issn.0578-1752.2014.17.016 Abstract （310）   HTML （3）    PDF （568KB）（464）       Knowledge map    Save 【Objective】 Genome-wide association studies were performed by using the chicken 60k high density SNP array for nine immune traits in Beijing-You chicken and identification of candidate genes and loci responsible for these traits were identified. Here, by treating Beijing-You chicken with polyinosinic acid-polycytidylic acid, Poly(I:C) and Salmonella enteritidis (SE), several candidate genes were further studied based on authors’ GWAS result, including CD1b, BMA1(B locus M alpha chain 1), TRIM27 (tripartite motif-containing 27) and ZNF692(zinc finger protein 692). 【Method】 Eighty 12-d-old Beijing-You Chickens were divided into three groups: the control group, Poly(I:C) treatment and SE treatment groups. All the experimental birds were reared in isolated facility. The treatment groups were, respectively, injected intramuscularly into the breast with 0.5 mL of Poly(I:C) and SE bacterial suspension containing 108 CFU and the control group was given 0.5 mL saline. Chickens were sacrificed at 12 h, 24 h, 3 and 6 days of post infection (DPI). Blood samples were collected and serum was stored. The bursa of Fabricius, thymus and spleen were rapidly removed to test inflammatory factors and gene expression. 【Result】 The weight was significantly lower than control group 24 h post infection (P＜0.01) and temperature of chickens was significantly changed in 24 h after treatments. In serum, concentrations of IFN-α, IL-4 and IL-6 were significantly higher than the control and reached a peak at 24 h or 3 d (P＜0.01). TNF-α increased in all periods and significantly higher than the control group after 3 days. The expression of candidate gene CD1b was not tissue-specific, but BMA1, TRIM27 and ZNF692 were highly expressed in thymus and bursa fabricius. In thymus, the expression of CD1b was significantly different between two treatments at 12 h and 24 h post infection. When treated with SE, the expression of CD1b was significantly increased and reached a peak at 24 h (P＜0.01). The mRNA expression of BMA1 was significantly different between two treatment groups in 12 h and 3 d post infection, when treated with Poly(I:C), the expression at 12 h post infection was lower than the SE group, but it was significantly higher than SE group (P＜0.01). The expression of TRIM27 on 6 DPI in Poly(I:C) treatment was significantly higher than control group, expression of ZNF692 was not different among three groups. In bursa of Fabricius, the expression of CD1b was the highest at 12 h after infection. There were no differences in the expression of genes BMA1 and TRIM27 between two treatments. The expression of ZNF692 was higher on 1 DPI in SE treatment than Poly(I:C), and also the highest on 3 DPI in both groups. 【Conclusion】 CD1b, BMA1,TRIM27 and ZNF692 might be involved in the immune response after treating with S. enteritidis and Poly(I:C). CD1b gene had an important role in the early stage of SE infections. BMA1 and ZNF692 were involved in immune response of thymus and bursa. Reference | Related Articles | Metrics

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Scientia Agricultura Sinica    2008, 41 (8): 2425-2430.   DOI: 10.3864/j.issn.0578-1752.2008.08.029 Abstract （1531）      PDF （330KB）（1000）       Knowledge map    Save Related Articles | Metrics

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Scientia Agricultura Sinica    2008, 41 (8): 2431-2435.   DOI: 10.3864/j.issn.0578-1752.2008.08.030 Abstract （1119）      PDF （384KB）（724）       Knowledge map    Save Related Articles | Metrics

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Scientia Agricultura Sinica    2008, 41 (8): 2436-2441.   DOI: 10.3864/j.issn.0578-1752.2008.08.031 Abstract （1332）      PDF （419KB）（769）       Knowledge map    Save Related Articles | Metrics

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Scientia Agricultura Sinica    2008, 41 (8): 2442-2447.   DOI: 10.3864/j.issn.0578-1752.2008.08.032 Abstract （1054）      PDF （392KB）（1024）       Knowledge map    Save Related Articles | Metrics

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Establishment of Real-time TaqMan-Fluorescence Quantitative RT-PCR Assay for Detection and Quantification of Porcine Lipoprotein Lipase mRNA Scientia Agricultura Sinica    2008, 41 (5): 1464-1469.   DOI: 10.3864/j.issn.0578-1752.2008.05.027 Abstract （1124）      PDF （499KB）（1254）       Knowledge map    Save bstract: Objective : Cloning porcine LPL cDNA , using it as the standard for real－time quantifying LPL mRNA and establishing TaqMan FQ-RT-PCR assay for detection of it. Methods : Total RNA extracted from longissimus dorsi of porcine was reverse transcribed to cDNA. LPL cDNA was ligated with pGM-T vector and transformed to bacterium TOP10. Plasmid DNA extracted from positive clones was verified by PCR amplification and sequenced. LPL was amplified by real-time fluorescence quantitative PCR from the plasmid DNA. The concentration of DNA template purified was detected by analysing absorbance in 260 nm and then was diluted to series standard concentrations of LPL αre combined plasmid for FQ-PCR. Results : The method of LPL mRNA real－time PCR was well established, which detected as low as 103 copies with the linear range from 103 to 1010 copies. The standard curves showed high correlations(R2=0.9871).Conclusion : A series of standards for real-time PCR analysis have been constructed successfully,and real-time TaqMan-Fluorescence Quantitative RT-PCR is reliable to quantitatively evaluate FQ-PCR mRNA in longissimus dorsi of porcine. Related Articles | Metrics

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Yak Lactate Dehydrogenase-A: Purification, Properties and cDNA Cloning and Sequencing Scientia Agricultura Sinica    2008, 41 (5): 1470-1475.   DOI: 10.3864/j.issn.0578-1752.2008.05.028 Abstract （1199）      PDF （507KB）（947）       Knowledge map    Save [Objective] In order to study the adaptation of yak (Bos. grunniens) to high altitude and low oxygen plateau at molecular level, lactate dehydrogenase A (LDH-A) from yak skeletal muscle was purified, and LDH-A cDNA was cloned. The kinetics and cDNA of yak LDH-A were compared with those of bovine LDH-A. [Methods] Dye affinity chromatography and DEAE-Sephadex ion-exchange chromatography were used to purify LDH-A from skeletal muscle of yak and bovine, and their enzyme properties were compared. The cDNA of yak LDH-A was cloned by RT-PCR methods and compared with that of bovine LDH-A in the GenBank. [Result] The relative activity of purified LDH-A was 103.9 U/mg protein, with purification factor of 18.2. Only one band was observed when the purified LDH-A was separated with SDS-PAGE or native PAGE. Kinetic analysis showed that Michaelis constants (Km) value for NADH was 0.097, and Km value for pyruvate was 1.897, both was higher than that of bovine. The Km value for pyruvate of yak LDH-A was about two folds that of bovine. Hg2+ inhibited LDH-A activity, and the inhibition effect was partly removed by adding β- mercaptoethanol. [Conclusion] The high Km for pyruvate of yak LDH-A can prevent from producing too much lactate in skeletal muscle, and might be a result of adaptive evolution.The two amino acid replacements are responsible for the increased Km value. Related Articles | Metrics

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The Preliminary Report on Rumen Protozoa Grazing Rate on Bacteria with a Fluorescence-labeled Technique WangMeng-zhi hongrong wang guoxiang li hengchun cao zhanjun lu Scientia Agricultura Sinica    2008, 41 (5): 1476-1481.   DOI: 10.3864/j.issn.0578-1752.2008.05.029 Abstract （1406）      PDF （308KB）（1050）       Knowledge map    Save Studies on the grazing rate of rumen protozoa were carried out under laboratory conditions using a technique of fluorescence-labeled bacteria, in Nov. 2006. Four Xuhuai goats were used in this experiment to provide rumen protozoa and bacteria. Two groups were designed as follows: one group was the whole bacteria which were marked using fluorescence through removing free bacteria from rumen fluid (WFLB); the other group was bacteria which were marked using fluorescence without removing free bacteria from rumen fluid (FLB). The result showed that, The grazing rates of rumen protozoa was 398.4cells/ (cell H) for the group with (WFLB), 230.4cells/ (cell H) for the group with (FLB); Conversed into bacteria-N, they were: 2.15pg N/ (cell H) for the group with (WFLB) and 1.24pgN/ (cell H) for the group with (FLB) respectively. Extrapolating the assimilation quantity of Nitrogen by ciliates on bacteria of Xuhuai goat, were 103.2 mg N/ (d capita) for the group with (WFLB), 59.5mg N/ (d capita) for the group with (FLB) respectively; It also could be estimated as protein loss, the protein micro-cycling were 0.645g Pr/ (d capita) for the group with (WFLB) and 0.372g Pr/ (d capita) for the group with (FLB) respectively, and finally fluorescence-labeled technique has a potential for research on the grazing rate of rumen protozoa. Related Articles | Metrics

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Effect of fucoidan and fraction isolated from Laminaria japonica on immunity of broiler macrophages under oxidation stress condition Scientia Agricultura Sinica    2008, 41 (5): 1482-1488.   DOI: 10.3864/j.issn.0578-1752.2008.05.030 Abstract （1250）      PDF （374KB）（870）       Knowledge map    Save Related Articles | Metrics

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Study on Dynamic Expression of the Apoptotic Gene-Fas/FasL in Immunity Organs of Gushi Chickens and its Physiological Significane LI Kui Ying LIU Gui-rong SUN Scientia Agricultura Sinica    2008, 41 (5): 1489-1496.   DOI: 10.3864/j.issn.0578-1752.2008.05.031 Abstract （1098）      PDF （233KB）（881）       Knowledge map    Save Abstract: The dynamic expression of the apoptotic gene-Fas/FasL in immunity organs of Gushi chickens was studied by using immunohistochemistry technology and Leica Microsystems, and its physiological significane was disscussed. The results showed that the expression of Fas in different immunity organs were observed, however, the number of the positive Fas-cells were different between different developmental stages. The positive Fas-protein was found in cell membrane, cytoplasm, but not in nucleus of lymphocytes. The positive Fas-cells were dispersed (or in clusters), and distributed in different places for different immunity organs. The majority of positive Fas-cells, in bursa, were observed in lamina propria of mucous membrane near epithelium and areas between lymphoid nodules, edges of lymphoid nodules, and a few in inner of lymphoid nodules. In thymus, the positive Fas-cells localized mainly in thymic medulla, and few in thymic cortex. The positive Fas-cells, in spleen, were found predominantly in red pulp, marginal zones, and the surronding areas of splenic nodules and periarterial lymphatic sheath, few in splenic nodules and periarterial lymphatic sheath. The expression of FasL in immunity organs of Gushi chickens is similar to Fas, but, the positive FasL-cells is less than Fas. All results mentioned above indicate that the apoptotic gene-Fas/FasL participates in the regulation of apoptosis of thymocytes and lymphocytes in immunity organs of Gushi chickens, and it is important for the stability of immunity organs. Related Articles | Metrics

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The expresstional differences and developmental changes of bcl-2、P53 Gene in early embryos of generic hybrids of chicken-quail Scientia Agricultura Sinica    2008, 41 (5): 1497-1502.   DOI: 10.3864/j.issn.0578-1752.2008.05.032 Abstract （1215）      PDF （514KB）（719）       Knowledge map    Save Abstract：【OBJECTIVE】To investigate the influences of apoptotic factors bcl-2、P53 on early embryos of generic hybrids;【METHOD】we acquired the cross bred eggs of chicken(♂)-quail(♀)with artificial insemination,hatched the eggs in one batch according to the standard condition of chicken,collected the early living-embryos of 2.75,3.0,3.25,3.50,3.75,4.0,4.25,4.5,4.75 and 5.0 d at random.Adopted the method of RT- PCR,used multi-ply PCR to identify the embryo sex with primers of Wpkci and β-actin,then selected four embryos of female and male respectively of each period,taken β-actin for internal standard to determine the relative quantity of bcl-2、P53 mRNA of embryos;【RESULTS】1)The bcl-2 mRNA expression of 2.75～4.75 d of male embryos maintained lower level,declined on 4.0 d,then reached the initial level,rose distinctly on 5.0 d(P<0.01)and reached the peak;the bcl-2 mRNA expression of 2.75～4.5 d of male embryos maintained lower level,rose distinctly on 4.75 d(P<0.01),then maintained higher level and reached the peak on 5.0 d;the female bcl-2 mRNA expression of 4.75 d were higher distinctly than male of the same days(P<0.05); 2)The patterns of P53 mRNA expression of male and female embryos were basically consistent.They were higher on 2.75 d,declined on 3.0 d(P<0.05)and reached the lowest level,declined distinctly in male,but the whole differences weren’t significant in female,then reached and maintained the initial level.Comparing the expression between different sex, the differences weren't significant.【CONCLUSION】We found two abnormal points in gene expresstional periods in female(3.0 and 4.75 d),but one point in male(3.0 d).The disorder regulated by key factors of apoptosis may induce embryo death.This conclusion should be confirmed further after the research of bcl-2、P53 expression of chicken and quail's embryos of the same period. Related Articles | Metrics

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