The Fluorescent Characterization and Analysis of Two Brucella Attenuated Vaccine Infecting Mouse Macrophagocyte     

HU Meng-wei  LIU Peng-tao  YAN Guo  GAO Jian-feng 《中国畜牧兽医》2016,43(8):1944-1950

The experiment was conducted to discuss the difference of binding time of green fluorescent protein B.melitensis M5 (GFP-M5) and B.abortus S19 (GFP-S19) infecting the mouse macrophagocyte (RAW264.7),lysosome,endoplasmic reticulum and golgi body in the initial stage and compare the binding rate of GFP-M5,GFP-S19 with organelle in different timeline,respectively,by confocal laser scanning microscope (CLSM) and flow cytometry.The result showed that GFP-M5 and GFP-S19 were successfully constructed.The intracellular survival ability of Brucella M5,Brucella S19,GFP-M5 and GFP-S19 were not obvisouly affected after infecting RAW264.7.GFP-M5 and GFP-S19 could enter the macrophagocyte in 30 mins,and in 2 h the Brucella could reach lysosome,endoplasmic reticulum and golgi body.In addition,the binding time for two attenuated vaccine did not show differences in 1,2,3 and 4 h.The content of GFP⁺ cell produced by RAW264.7 infected by GFP-M5 and GFP-S19 did not show significant differences (P>0.05).Therefore,the two strains did not have significant differences in the invasion ability in the initial stage of infecting host cell.  相似文献   

Effect of Se and VE Supplement on Semen Quality,Antioxidant Enzyme Activities and Heat Shock Protein Expression of Goat in High Temperature Season     

SHI Li-guang  ZHOU Xiong  XUN Wen-juan  HOU Guan-yu  ZHOU Han-lin  HUANG Xian-zhou 《中国畜牧兽医》2016,43(1):101-107

This experiment was conducted to study the effect of selenium and vitamin E supplement on semen quality, antioxidant enzyme activities and heat shock protein expression of goat in Hainan high temperature season.16 adult Hainan Black goat with good health and approximate weight were randomly divided into 4 groups, fed with basal diet(control group), basal diet+0.5 mg/kg Se(Se group), basal diet+100 mg/kg VE(VE group), and basal diet+0.5 mg/kg Se+100 mg/kg VE(Se+VE group), respectively.The experimental period was 93 d.Semen samples were collected in the last week of the experiment on two consecutive days.The semen quality, antioxidant enzyme activities and heat shock protein expression were analyzed.The results showed that compared with control group, the ejaculate volume was not significantly affected by Se or VE supplement(P>0.05).Sperm density and sperm motility were increased significantly by Se and VE supplement(P<0.05), and the abnormal rate was decreased extremely significantly(P<0.01).The goats fed with Se and VE also had higher activities of GSH-Px(P<0.01), SOD(P<0.05), CAT(P<0.05) and T-AOC(P<0.01), and lower MDA concentration in seminal plasma(P<0.05).The relative expression levels of HSP70 and HSP90 mRNA in supplement groups were decreased extremely significantly(P<0.01).However, there were some certain differences between the Se and VE supplement groups on semen quality and heat shock protein expression.In conclusion, the supplementation of Se and VE could help to improve goat semen quality by increasing the sperm density, sperm motility, the antioxidant enzyme activities, and decreasing the abnormal rate in hot season of Hainan.Finally, Se and VE supplement had good effects on relieving the environment heat stress.  相似文献   

Effects of Dietary Crude Protein Levels on Growth Performance,Digestion and Metabolism,and Serum Biochemical Indexes of Super Merino Lamb     

WANG Chun-xin  ZHAO Yun-hui  ZHAO Zhuo  ZHU Jing-liang  ZHAI Bo  ZHAO Feng-jiang  ZHANG Ming-xin 《中国畜牧兽医》2016,43(1):108-113

The experiment aimed to find out the effects of dietary crude protein levels on growth performance, digestion and metabolism and serum biochemical indexes of Super Merino lamb after weaning.According to the dietary crude protein level, 64 Super Merino weaned lambs were divided into four groups, which the crude protein levels were 13.25%(group Ⅰ), 14.33%(group Ⅱ), 15.53%(group Ⅲ) and 16.60%(group Ⅳ), respectively.The experiment lasted for 60 days, 30 days for the early stage and 30 days for the later stage;And on the 30th and 60th day, blood samples were collected from one head sheep chosen from each replication, and at the end of the feeding test, 3 lambs were randomly selected from each experiment group for a 15 days digestion and metabolism experiment.The results showed that ADFI and ADG in the early and total trial period were significant differences(P<0.05), ADFI and ADG of group Ⅲ were higher than others.The apparent digestibility of nutrients and ME/DE were not significantly changed(P>0.05).In the detection of serum biochemical indexes, there was no significant difference in the earlier stage(P>0.05);At the end of the test, PK and CK showed significant difference between groups(P<0.05).PK presented a rising trend as the protein level improved, CK of group Ⅰ was higher than other groups.In Super Merino lamb early weaning diet, 15.53% protein level in the diet could significantly improve ADG, the apparent digestibility of nutrients, metabolic energy digestibility.At the end of the test, PK and CK were affected by dietary crude protein level significantly.  相似文献   

Prokaryotic Expression and Antigenicity Analysis of Porcine Japenese Encephalitis Virus Envelope Protein Domain Ⅲ     

HOU Hua-yan  DING Ling-ling  LV Mao-jie  YANG Bao-shou 《中国畜牧兽医》2016,43(4):934-939

In order to analyze the antigenicity of porcine Japanese encephalitis virus (JEV) E protein domain Ⅲ, which was expressed by pET-28a vector with His-tag and purified through Ni-NTA, the BALB/c mice were immunized with the purified protein.We identified the antigenicity of domain Ⅲ of E protein and the anti-mice and anti-porcine JEV E Ⅲ protein specific antibody titers by SDS-PAGE, Western blotting, indirect ELISA and IFA.SDS-PAGE results showed the expressed target protein existed mainly in the form of inclusion body.Western blotting, ELISA test results showed that the protein had good reactivity with anti-serum.The mice immunized with the purified JEV E Ⅲ protein generated 1×10⁵ anti-JEV E Ⅲ protein specific antibody titers by ELISA, and the porcine immunized with the porcine JEV generated 5.1×10⁴ anti-JEV specific antibody titers.The IFA results showed that JEV E Ⅲ protein anti-serum could identify JEV antigen.The above results showed that the recombinant JEV E Ⅲ had good antigenicity.These results provided important basis for development of diagnostic antigen for JEV.  相似文献   

Effects of Complex Nutritional Regulation Additives on Growth Performance,Slaughter Performance and Serum Biochemical Indexes of Grazing Sheep in Cold Season     

LIANG Jing  ZHANG Wen-ju  WANG Bo 《中国畜牧兽医》2016,43(4):953-959

This experiment was conducted to investigate the effects of complex nutritional regulation additives on growth performance, slaughter performance and serum biochemical indexes of grazing sheep in cold season.A single factor random block design was used and 48 grazing ewes with the average weight (28.01±1.70) kg and 7 months old were randomly divided into 4 groups:Group Ⅰ without supplement;Group Ⅱ with concentrate;Group Ⅲ with concentrate+complex formulation 1 (0.30% malic acid, 0.06% cysteamine, 0.08% saccharicterpenin);Group Ⅳ with concentrate+complex formulation 2 (0.60% malic acid, 0.12% cysteamine, 0.16% saccharicterpenin)with 3 replicates per group and 4 grazing ewes per replicate.The results showed that:①The final weight of group Ⅲ was the highest and higher than group Ⅱ and Ⅳ by 6.26% (P>0.05)、3.69% (P>0.05), respectively.The average daily gain of group Ⅲ was highest, and higher than group Ⅱ and Ⅳ by 26.32% (P<0.01)、5.74% (P>0.05) respectively.②Comparing with group Ⅱ, the slaughter rate of group Ⅲ was increased by 4.68% (P>0.05).Compared with groupⅠ, Ⅱ and Ⅳ, the meat percentage of group Ⅲ was increased 18.46% (P<0.05), 10.97% (P>0.05) and 4.40% (P>0.05), while the net meat weight of carcass in group Ⅲ was increased by 49.74% (P<0.01), 13.22% (P>0.05) and 18.27% (P<0.05), respectively.Compared with groupⅠ, Ⅱ and Ⅳ, the net meat rat of carcass in group Ⅲ was increased 10.42% (P<0.01)、6.00% (P>0.05) and 3.37% (P>0.05), respectively.③There was a rising trend as the order of groupⅠ, Ⅱ, Ⅳ and Ⅲ in serum glucose, Ca, P, HDL, LDL, GPT and IgG, while A/G and TG showed a decreasing trend.Based on the above results, in this trial, supplementing complex nutritional regulation additives had good effects on growth performance, slaughter performance and serum biochemical indexes of grazing sheep in cold season.And supplementing complex formulation group 1 (group Ⅲ) was the best.  相似文献   

多型FMDV VP1表位嵌入型载体pMG-SLPMultiVP1的构建及其表达产物鉴定     

陆继爽  黄天鹏  吴咪  贺海燕  格日勒图 《动物医学进展》2016,(10):6-10

构建出表达载体pMG-SLPMultiVP1,并对其进行双酶切鉴定及PCR鉴定。将阳性重组质粒转入到发酵乳杆菌中进行表达,并对表达产物进行SDS-PAGE、Western blot和IFAT鉴定。多型FMDV VP1表位嵌入的SLPMultiVP1融合基因成功克隆入穿梭质粒pMG36e中,其表达产物大小约为35ku,部分表达产物位于发酵乳杆菌表面。成功构建了一个乳杆菌嵌入型表达载体pMG-SLPMultiVP1,为以发酵乳杆菌为活菌载体的表位疫苗研究奠定了基础。  相似文献   

内质网应激偶联炎性反应研究进展     

廖轶  崔永勇  刘春法  杨利峰  赵德明  周向梅 《动物医学进展》2016,(7):89-93

内质网是真核细胞蛋白合成和折叠的主要细胞器,当内质网内蛋白合成或折叠负担增加,引起未折叠或错误折叠蛋白增多时,可激活内质网几条特定信号通路,启动未折叠蛋白反应,这对维持细胞稳态有重要意义。越来越多研究表明,多种炎性反应疾病与内质网应激有密切联系。一方面,内质网应激引起的未折叠蛋白反应可以诱发或者抑制炎症,另一方面,炎性反应也能影响蛋白折叠,从而促进或缓解内质网应激。2型糖尿病、肿瘤和动脉粥样硬化等多种重大疾病的病理机制都涉及内质网应激与炎性反应的相互作用,对该问题的深入研究不仅能加深人们对这些疾病发病机制的理解,也有助于相关药物的研发。  相似文献   

噬菌体Bp7尾丝蛋白gp37的原核表达与鉴定     

孙虎芝  任慧英  张灿 《动物医学进展》2016,(5):10-14

为了研究噬菌体尾丝蛋白gp37在识别宿主菌大肠埃希菌K12过程中的作用,PCR扩增获得gp37基因C端1 161bp序列,以EGFP为荧光标签,构建重组表达质粒pET-28a-EGFP-gp37,在大肠埃希菌BL21(DE3)中IPTG诱导表达重组蛋白EGFP-gp37,亲和层析纯化蛋白。SDS-PAGE及Western blot检测结果表明,重组蛋白EGFP-gp37在大肠埃希菌BL21(DE3)为可溶性表达,表达量占菌体总蛋白量的23%。EGFP-gp37融合蛋白与大肠埃希菌K12相互作用,激光共聚焦显微镜检测结果显示,尾丝蛋白gp37能够吸附到宿主菌K12表面,表明尾丝蛋白gp37参与宿主菌受体的识别和吸附过程。  相似文献   

Effects of water‐misting sprays with forced ventilation on post mortem glycolysis,AMP‐activated protein kinase and meat quality of broilers after transport during summer          下载免费PDF全文

Nannan Jiang  Tong Xing  Minyi Han  Shaolin Deng  Xinglian Xu 《Animal Science Journal》2016,87(5):718-728

Effects of water‐misting sprays with forced ventilation on post mortem glycolysis, adenosine monophosphate‐activated protein kinase (AMPK) and meat quality of broilers after transport during summer were investigated in the present paper. A total of 105 mixed‐sex Arbor Acres broilers were divided into three treatment groups: (i) 45 min transport without rest (T); (ii) 45 min transport with 1 h rest (TR); and (iii) 45 min transport with 15 min water‐misting sprays with forced ventilation and 45 min rest (TWFR). Each treatment consisted of five replicates with seven birds each. The results indicated that the water‐misting sprays with forced ventilation could mitigate the stress caused by transport under high temperature conditions during summer, which reduced the energy depletion in post mortem Pectoralis major (PM) muscle. This resulted in a higher energy status compared to the T group, which would decrease the expression of phosphorylation of AMPK (p‐AMPK). Furthermore, decreased the expression of p‐AMPK then slowed down the rate of glycolysis in post mortem PM muscle during the early post mortem period, which in turn lessened the negative effects caused by transport on meat quality. In conclusion, water‐misting sprays with forced ventilation may be a better method to control the incidence of the pale, soft and exudative meat in broilers.  相似文献   

The long-term effects of fire regime on primary production of montane grasslands in South Africa     

Colin S Everson  Terry Everson 《African Journal of Range and Forage Science》2016,33(1):33-41

Natural grasslands deliver essential ecosystem services through plant production, which enhances water supply, nutrient cycling, soil retention and greenhouse gas mitigation. Although the condition of montane grasslands for provision of ecosystem services is maintained by regular annual or biennial burning, controversy exists over the impact of different frequencies and seasons of burning on grassland productivity. The objective of this study was to determine the long-term effects of different burning regimes on primary production and quality of the montane grasslands of the KwaZulu-Natal Drakensberg. There were no significant differences in the mean standing live mass between 30 years of annual winter and biennial spring burning. However, in unburnt areas productivity was 20% lower (118.2?g m?2) than in regularly burnt grassland (144.7–154.5?g m?2). Crude protein did not vary between the annual winter and biennial spring treatments (95–113?kg ha?1), but was significantly lower in unburned areas (45?kg ha^?1). However, an infrequent fire in a protected area caused a temporary spike in crude protein (16%) compared with regular burning (5–10%), which can benefit wildlife. We conclude that montane grasslands can be burnt annually or biennially in the dormant season to promote long-term productivity.  相似文献