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一株猪流感病毒A/swine/Shanghai/01/2019(H1N1)的分离鉴定及遗传进化分析 被引量:8
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作者 程靖华 陶洁 +2 位作者 李本强 石迎 刘惠莉 《中国预防兽医学报》 CAS CSCD 北大核心 2020年第8期830-834,共5页
为鉴定上海地区猪流感病毒的流行株及其分子生物学特征,本研究从上海地区养殖场采集疑似流感症状的猪咽拭子样品40份,采用套式PCR结合鸡胚分离鉴定方法,从中分离到1株H1N1亚型猪流感病毒(SIV),命名为A/swine/Shanghai/01/2019(H1N1)。... 为鉴定上海地区猪流感病毒的流行株及其分子生物学特征,本研究从上海地区养殖场采集疑似流感症状的猪咽拭子样品40份,采用套式PCR结合鸡胚分离鉴定方法,从中分离到1株H1N1亚型猪流感病毒(SIV),命名为A/swine/Shanghai/01/2019(H1N1)。经全基因组测定和遗传进化分析结果显示该分离株的8个基因节段与欧亚类禽H1N1 SIV同源性最高,其PB2、PB1、PA基因节段可能来源于A/swine/Jiangsu/49/2012(H1N1)病毒,NA、NP和M基因节段可能来源于A/swine/Shanghai/3/2014(H1N1)病毒。该分离病毒的HA蛋白裂解位点为PSIQSR↓GLFGAI,为低致病性流感病毒的分子特征。该分离株NA蛋白未出现与神经氨酸酶抑制剂相关的氨基酸突变,M2蛋白S31N突变表明该病毒可能对金刚烷胺类药物具有一定的耐药性。PA蛋白224S和PB2蛋白701N突变提示该分离株对哺乳动物的适应性增强,可能导致其致病性增强,需要加强对此类SIV的监测。结合近年来的监测表明,上海地区猪群中H1N1亚型SIV持续存在,需要持续跟踪监测SIV变异趋势。本研究为防控猪流感提供实验依据。 展开更多
关键词 猪流感病毒 H1N1亚型 分离鉴定 遗传进化分析
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Incorporation of meat quality and use of ultrasound technology for swine genetic improvement programs 被引量:9
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作者 Tom J.BAAS 《华南农业大学学报》 CAS CSCD 北大核心 2005年第S1期88-98,共11页
Real-time ultrasound has been, and will continue to be, one of the most important tools in enhancing genetic improvement in the U.S. Recent advancements in the technology have allowed ultrasound to be used in evaluati... Real-time ultrasound has been, and will continue to be, one of the most important tools in enhancing genetic improvement in the U.S. Recent advancements in the technology have allowed ultrasound to be used in evaluating intramuscular fat in live animals. Together with other genetic improvement tools, ultrasound technology will offer seedstock producers the opportunity to select for improved IMF in potential breeding stock replacements and hence speed genetic progress for the improvement of this trait. After three generations of selection for IMF using real-time ultrasound in an Iowa State University study, the average EBV for select line pigs is 0.83% greater than for control line pigs. Selection for IMF has, however, resulted in slightly more backfat and less loin muscle area, and a trend toward more days to 114 kg in the select line compared to the control line. Carcass evaluation of a sample of pigs from each litter indicated a similar increase in IMF, increase in backfat, and reduction in loin muscle area for select line pigs. No differences were found for Hunter L* color, Minolta reflectance, and ultimate pH. 展开更多
关键词 swine meat quality intramuscular fat ultrasound technology
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人源H3N2亚型猪流感病毒A/Swine/Guangdong/1/2005的分离鉴定及全基因遗传演化分析 被引量:2
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作者 许传田 董国英 +2 位作者 杨焕良 乔传玲 陈化兰 《中国预防兽医学报》 CAS CSCD 北大核心 2009年第4期322-324,共3页
2005年从广东某猪场采集疑似流感病猪的病料(气管和肺脏)样品共40份,通过MDCK细胞培养分离到6株A型流感病毒,经过血凝抑制和PCR扩增方法鉴定均为H3N2亚型。挑选其中一个代表株A/swine/Guangdong/1/05(H3N2)(SwGD1/05)进行全基因组测序并... 2005年从广东某猪场采集疑似流感病猪的病料(气管和肺脏)样品共40份,通过MDCK细胞培养分离到6株A型流感病毒,经过血凝抑制和PCR扩增方法鉴定均为H3N2亚型。挑选其中一个代表株A/swine/Guangdong/1/05(H3N2)(SwGD1/05)进行全基因组测序并与GenBank中相关序列比较,结果发现SwGD1/05的8个基因分别与人流感病毒相关基因氨基酸同源性高达98.5%~99.4%;SwGD1/05的8个基因进化树中分别位于北美和欧洲人流感病毒进化谱系位置;由此推测SwGD1/05可能来源于北美或欧洲人源流感病毒株。这一结果为阐明猪作为流感病毒传播的中间宿主作用提供了的理论支持,本研究具有重要的人类公共卫生意义。 展开更多
关键词 人源猪流感病毒 H3N2亚型 分离鉴定 遗传演化
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H1N2亚型猪流感病毒A/Swine/Guangdong/1/06分离鉴定和对猪致病性研究 被引量:2
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作者 许传田 樊树芳 +4 位作者 杨焕良 乔传玲 陈艳 辛晓光 陈化兰 《中国预防兽医学报》 CAS CSCD 北大核心 2009年第1期46-50,共5页
2006年5月从广东某大型猪场采集具有流感症状保育猪鼻拭子共98份,无菌常规处理猪鼻拭子后接种MDCK细胞,分离到4株流感病毒。用血凝抑制和PCR方法鉴定均为H1N2亚型。挑选其中一株A/Swine/Guangdong/1/06(H1N2),进行全基因组测序,并与GenB... 2006年5月从广东某大型猪场采集具有流感症状保育猪鼻拭子共98份,无菌常规处理猪鼻拭子后接种MDCK细胞,分离到4株流感病毒。用血凝抑制和PCR方法鉴定均为H1N2亚型。挑选其中一株A/Swine/Guangdong/1/06(H1N2),进行全基因组测序,并与GenBank中相关序列进行比较。核苷酸同源性分析表明:A/Swine/Guangdong/1/06(H1N2)与A/Swine/Guangxi/13/06不同基因之间同源性为96.6%~98.1%。以106EID50的剂量,将H1N2病毒鼻腔感染35日龄仔猪,结果表明H1N2亚型流感病毒可以感染猪上呼吸道,但不表现临床症状。本研究结果对于揭示H1N2亚型猪流感流行规律和病毒的致病机理具有一定的意义。 展开更多
关键词 H1N2亚型猪流感病毒 分离鉴定 致病性
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猪流感病毒A/Swine/Inner Mongolia/547/01分离株神经氨酸酶基因的克隆及真核表达 被引量:3
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作者 赵朴 李海燕 +3 位作者 杨焕良 陈化兰 于康震 卢中华 《中国预防兽医学报》 CAS CSCD 北大核心 2005年第1期1-4,共4页
根据GenBank中登录的猪流感病毒参照序列设计了扩增神经氨酸酶 (NA)基因的特异性引物。从猪流感病毒A/Swine/InnerMongolia/5 4 7/0 1(H3N2 )株感染的鸡胚液中直接提取病毒RNA ,经RT_PCR扩增后将其克隆到pMD18_T载体上 ,进行序列测定和... 根据GenBank中登录的猪流感病毒参照序列设计了扩增神经氨酸酶 (NA)基因的特异性引物。从猪流感病毒A/Swine/InnerMongolia/5 4 7/0 1(H3N2 )株感染的鸡胚液中直接提取病毒RNA ,经RT_PCR扩增后将其克隆到pMD18_T载体上 ,进行序列测定和拼接。用HindⅢ酶切后 ,亚克隆到杆状病毒转移载体pMelBacB上 ,得到重组转移载体pMelBacNA ,然后与线性化杆状病毒DNA(Bac_N_BlueTMDNA)共转染于sf9昆虫细胞。经 3轮噬斑纯化 ,提取其DNA经PCR鉴定 ,获得重组杆状病毒株。SDS_PAGE结果显示 ,目的基因已获得表达 ,Westernblot和神经氨酸酶活性检测结果显示表达产物具有良好的免疫原性和神经氨酸酶活性 。 展开更多
关键词 猪流感病毒 神经氨酸酶 基因克隆 基因表达 流感
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猪流感病毒(A/swine/Jiangsu/2/2006(H3N2))NS1基因表达产物分析
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作者 郭林 王晓杜 +5 位作者 刘庆伟 沈阳 邱亚峰 李向东 罗满林 马志永 《中国动物传染病学报》 CAS 2009年第1期22-28,共7页
采用RT-PCR方法克隆猪流感病毒H3N2亚型NS1全长基因,构建NS1基因原核表达质粒pET-28a-NS1和真核表达质粒p3xFLAG-CMV-NS1,在大肠杆菌和真核细胞内表达NS1基因,制备抗NS1多克隆抗体。用所制备的抗体分析p3xFLAG-CMV-NS1转染表达NS1蛋白... 采用RT-PCR方法克隆猪流感病毒H3N2亚型NS1全长基因,构建NS1基因原核表达质粒pET-28a-NS1和真核表达质粒p3xFLAG-CMV-NS1,在大肠杆菌和真核细胞内表达NS1基因,制备抗NS1多克隆抗体。用所制备的抗体分析p3xFLAG-CMV-NS1转染表达NS1蛋白和病毒感染细胞内的NS1蛋白。经终浓度为1mmol/L的IPTG诱导后,重组蛋白NS1在大肠杆菌中得到表达。表达蛋白纯化后免疫Wistar大鼠制备抗NS1蛋白多克隆抗体,Western-blot分析表明抗NS1多克隆抗体可以识别大肠杆菌表达的NS1蛋白、转染Vero细胞表达的NS1蛋白和病毒感染细胞内的NS1蛋白。间接免疫荧光发现NS1蛋白主要定位于细胞核。结果显示,本试验成功构建了NS1基因原核和真核表达系统,获得了NS1特异性多克隆抗体,分析了NS1细胞定位,为进一步研究NS1蛋白在病毒复制过程中的生物学功能和猪流感病毒的复制机理奠定基础。 展开更多
关键词 猪流感病毒 NS1基因 基因克隆 基因表达 抗体制备
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Establishing an ELISA,Based on the HA1 Protein,for Detecting Antibodies against H3N2 Subtype Swine Influenza Virus
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作者 DING Xuan-ya QIAO Chuan-ling +4 位作者 CHEN Yan YANG Huan-liang XIN Xiao-guang HAN Qing-gong CHEN Hua-lan 《畜牧兽医学报》 CAS CSCD 北大核心 2009年第S1期57-61,共5页
The HA1 gene of H3N2 subtype swine influenza virus(SIV)was cloned into the expression plasmid pET-30a,the recombinant plasmid was named pET-HAl.This was transformed into E.coli BL21(DE3),and expressed by induction wit... The HA1 gene of H3N2 subtype swine influenza virus(SIV)was cloned into the expression plasmid pET-30a,the recombinant plasmid was named pET-HAl.This was transformed into E.coli BL21(DE3),and expressed by induction with IPTG.The expressed HA protein was identified by SDS-PAGE and Western blotting which showed the protein to be 42kDa and was immunoreactive.The purified HA protein was used to establish the indirect ELIS A for detection of the antibodies,specifically against the H3 subtype of SIV.The assay has excellent specificity,sensitivity and reproducibility.When 96 serum samples,randomly collected from the field,were evaluated in parallel by this new ELISA using recombinant HA1 and a routine HI test,the coincidental rate between the two tests was 86.5%.These results show that the recombinant HAl-based ELISA is specific,sensitive and easy to perform for the serological diagnosis of SIV infection. 展开更多
关键词 swine INFLUENZA virus HA1 PROTEIN SUBTYPE specific ELISA
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Phylogeny and Homologous Recombination Occurring in Classical Swine Fever Viruses
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作者 Jiang Qing-yun Gao Yu +4 位作者 Li Wei-qun Ding Fan Juergen Richt Ren Yu-dong Li Guang-xing 《Journal of Northeast Agricultural University(English Edition)》 CAS 2019年第3期57-65,共9页
Classical swine fever virus(CSFV) is the causative agent of classical swine fever, a highly contagious disease of pigs. But there is little information on the recombination in natural populations of CSFVs. Therefore, ... Classical swine fever virus(CSFV) is the causative agent of classical swine fever, a highly contagious disease of pigs. But there is little information on the recombination in natural populations of CSFVs. Therefore, a phylogenetic analysis of 62 fulllength genome CSFV strains, isolated from all over the world, was performed to detect potential recombination events, with the recombinant sequences being analyzed with the SimPlot and RDP programs. The results identified a mosaic virus, Chinese CSFV HCLV(2)(AF091507.1), which is the one naturally emerged recombinant CSFV with two recombination breakpoints at 2 484 and 2 900 bp of the genome alignment. Its two putative parental-like strains were CSFV Shimen(AF092448.2) and CSFV strain C/HVRI(AY805221.1). This work demonstrated that homologous recombination did occur in natural CSFV populations. It had significant implications for understanding the molecular epidemiology of CSFV, and revealed that recombination was an important factor for high genetic diversities of CSFV. 展开更多
关键词 CLASSICAL swine FEVER virus HOMOLOGOUS recombination PHYLOGENY genetic diversity
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Construction and Immunogenicity of a Recombinant Adenovirus Expressing the HA Gene of H5N1 Subtype Swine Influenza Virus
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作者 WU Yun-pu QIAO Chuan-ling +4 位作者 YANG Huan-liang CHEN Yan ZHAN Xiao-guo XIN Xiao-guang CHEN Hua-lan 《畜牧兽医学报》 CAS CSCD 北大核心 2010年第S1期76-81,共6页
To construct a recombinant adenovirus shuttle plasmid pDC315-H5HA-EGFP,the HA gene of A/Swine/Fujian/1/2001(H5N1) was amplified by RT-PCR and then inserted into adenovirus shuttle plasmid pDC315.A replication-defectiv... To construct a recombinant adenovirus shuttle plasmid pDC315-H5HA-EGFP,the HA gene of A/Swine/Fujian/1/2001(H5N1) was amplified by RT-PCR and then inserted into adenovirus shuttle plasmid pDC315.A replication-defective recombinant adenovirus expressing the HA gene(rAd-H5HA-EGFP) was generated by co-transfecting the recombinant shuttle plasmid pDC315-H5HA-EGFP and the genomic plasmid pBHGlox△E1,E3Cre in HEK293 cells.The recombinant adenovirus was confirmed by PCR,RT-PCR and Western blot assay.These results demonstrated that HA protein was properly expressed by the rAd-H5HA-EGFP in HEK293 cells and had natural biological activities.The TCID<sub>50</sub> of the rAd-H5HA- EGFP was assessed to be 2.26×10<sup>10</sup>/mL after propagation and purification.Immunization of BALB/ c mice indicated that rAd-H5HA-EGFP induced HI antibodies and protected mice from replication of the challenge virus in their lungs. 展开更多
关键词 swine influenza virus H5N1 subtype HA gene recombinant adenovirus
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National Swine Genetic Improvement: An overview of essential program components and organizational structure needed for success
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作者 John MABRY 《华南农业大学学报》 CAS CSCD 北大核心 2005年第S1期20-27,共8页
The swine industry in China is a thriving and evolving industry that has shown phenomenal growth over the past 10 years. To insure long term success and viability in a worldwide competitive industry such as pork, ther... The swine industry in China is a thriving and evolving industry that has shown phenomenal growth over the past 10 years. To insure long term success and viability in a worldwide competitive industry such as pork, there is need for a National Swine Genetic Improvement Program. This program needs to draw on expertise and technology from across the world for its development, but it should be based on the structure of the pig industry in China and be led by Chinese scientists, administrators and producers. National Genetic Improvement requires more than just technology. A successful program of national genetic improvement will require cooperation from the industry and the government. The support for the university system is essential for the success of the pig industry. The university system has a vital role on education (of students, faculty, producers and consumers) as well as research and technology transfer. The government could also have a role in supporting the central test stations and AI stations across the country. An accurate and comprehensive pedigree maintenance system is essential to genetic improvement. And it will be vitally important to be active in the importation of new genetics to sample other populations. 展开更多
关键词 swine National Genetic Improvement Program organizational structure
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Developments in quantitative genetics methodology as applied to national genetic improvement programs for swine
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作者 Ignacy MISZTAL 《华南农业大学学报》 CAS CSCD 北大核心 2005年第S1期47-56,共10页
Genetic selection in pigs through BLUP was very successful. However, strong selection mainly on growth and number of born alive decreased fitness and reduced environmental changes that animals can tolerate especially ... Genetic selection in pigs through BLUP was very successful. However, strong selection mainly on growth and number of born alive decreased fitness and reduced environmental changes that animals can tolerate especially under suboptimal environments. Additional challenges are genetic differences between purebreds (selected animals) and crossbreds (commercial animals), and possibly different environments for these groups of animals. A successful genetic selection at this time requires comprehensive data for all levels of the pyramid, multitrait models for a variety of traits including categorical and survival, and software that can implement complicated models while supporting large data sets. Many projects in pig genetic evaluation are carried out at the University of Georgia. Those studies are supported by software family called BGF90. 展开更多
关键词 quantitative genetics breeding software national swine breeding program
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Eukaryotic Expression and Activity Analysis of Capsid Gene of Swine Vesicular Disease Virus HK/70
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作者 TIAN Hong WU Jingyan SHANG Youjun YING Shuanghui ZHENG Haixue LIU Xiangtao XIE Qingge 《Journal of Northeast Agricultural University(English Edition)》 CAS 2009年第4期25-30,共6页
The capsid protein precursor (P1), which plays a major role for the generation of polypeptides of swine vesicular disease virus (SVDV), was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and e... The capsid protein precursor (P1), which plays a major role for the generation of polypeptides of swine vesicular disease virus (SVDV), was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and expressed in the mammalian cell line PK15 through the retroviral expression system. The activity of recombinant protein to induce immune response was evaluated in guinea pigs. IFA and Western Blot were used to detect the recombinant protein expression. The results showed that the recombinant protein could be recognized by SVDV positive serum, and animal test showed SVDV-specific antibodies. All of those results indicate that a retroviral-based vaccine carrying the capsid protein precursor (P1) of SVD is able to be expressed in the eukaryotic cell and elicites strong SVDV-specific immune responses in guinea pigs. 展开更多
关键词 swine vesicular disease virus capsid protein precursor gene gene expression immune response
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Establishment of mitral regurgitation swine model and study the change of hydrogen sulfide system
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作者 Shangyu Liu Li Zheng +1 位作者 Yue Tang Yan Yao 《中国循环杂志》 CSCD 北大核心 2018年第S01期122-122,共1页
Objective To establish the mitral regurgitation swine model and study the change of hydrogen sulfide(H2S)system in chronic heart failure model.Methods Miniature pigs were randomly divided into two groups,the control g... Objective To establish the mitral regurgitation swine model and study the change of hydrogen sulfide(H2S)system in chronic heart failure model.Methods Miniature pigs were randomly divided into two groups,the control group(n=6)and mitral regurgitation group(n=6).Chronic heart failure models were established by pulling mitral chordal through a small incision extracorporeal. 展开更多
关键词 MITRAL REGURGITATION swine MODEL the CHANGE of hydrogen sulfide(H2S)system heart failure MODEL
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H5N1亚型猪流感病毒A/Swine/Fujian/1/01感染性克隆构建及其对小鼠致病性研究 被引量:2
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作者 郭威 许传田 +2 位作者 朱启运 陈海平 莫内 《中国预防兽医学报》 CAS CSCD 北大核心 2009年第5期400-402,共3页
2001年从福建某猪场分离到1株H5N1亚型猪流感病毒(SIV)A/Swine/Fujian/1/01(SW/FJ/1/01)。SW/FJ/1/01对小鼠具有强致病性,致死率为100%,为进一步研究SW/FJ/1/01对BALB/c小鼠致病性的分子基础,本研究构建了SW/FJ/1/01的8个节段重组质粒... 2001年从福建某猪场分离到1株H5N1亚型猪流感病毒(SIV)A/Swine/Fujian/1/01(SW/FJ/1/01)。SW/FJ/1/01对小鼠具有强致病性,致死率为100%,为进一步研究SW/FJ/1/01对BALB/c小鼠致病性的分子基础,本研究构建了SW/FJ/1/01的8个节段重组质粒构成的反向基因操作系统,成功拯救了病毒(R-SW/FJ/1/01)。R-SW/FJ/1/01和野生型SW/FJ/1/01对BALB/c小鼠致病性没有差别。SW/FJ/1/01反向基因操作系统的建立为进一步阐明H5N1亚型SIV对哺乳动物模型BALB/c小鼠的致病机制等方面的研究奠定了基础。 展开更多
关键词 H5N1猪流感病毒 反向遗传 哺乳动物模型
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猪流感病毒A/swine/Guangdong/2/2012(H1N1)毒株攻毒BALB/c小鼠细胞因子动态变化的研究 被引量:1
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作者 林文 吕艳丽 +6 位作者 黄良宗 刘远佳 唐志玲 余绍华 刘明杰 罗满林 顾万军 《中国畜牧兽医》 CAS 北大核心 2013年第10期11-16,共6页
为了测定H1N1亚型猪流感病毒(swine influenza virus,SIV)对小鼠的致病性,本试验对A/swine/Guangdong/2/2012(H1N1)株SIV HA基因进行克隆及遗传分析,并将SIV尿囊液经鼻腔感染6周龄BALB/c小鼠,观察感染后小鼠的一般状况、器官系数和组织... 为了测定H1N1亚型猪流感病毒(swine influenza virus,SIV)对小鼠的致病性,本试验对A/swine/Guangdong/2/2012(H1N1)株SIV HA基因进行克隆及遗传分析,并将SIV尿囊液经鼻腔感染6周龄BALB/c小鼠,观察感染后小鼠的一般状况、器官系数和组织病理学变化,在病毒感染后第1、3和7天使用荧光定量PCR测定小鼠肺脏、脾脏、脑组织中7种细胞因子mRNA的表达量,研究其对小鼠的致病特性。结果显示,该病毒属于经典SIV,病毒经鼻腔感染后可引起小鼠活动减少、采食量降低,但无咳嗽和死亡;病理组织学变化为肺间隔较正常组织明显增厚,毛细血管明显扩张充血,周围肺泡腔呈代偿性肺气肿;小鼠肺脏、脾脏组织样本中IFN-α、IFN-β、IP-10、IL-1β、TNF-α、IRF-3和IL-10mRNA含量在感染后第3天均显著升高(P<0.05),而脑组织样本中IL-1β和IL-10在小鼠攻毒后第3和7天均显著上调(P<0.05)。 展开更多
关键词 H1N1亚型猪流感病毒 BALB C小鼠 细胞因子
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Primary Selection of MicroRNA Acting on Classical Swine Fever Virus in Swine Umbilical Vein Endothelial Cells
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作者 HOU Bo ZHANG Yan-ming NING Peng-bo KANG Kai DENG Li DAI Chen rANG Qing-hai 《畜牧兽医学报》 CAS CSCD 北大核心 2011年第B12期30-33,共4页
The aim of the study was to identify the microRNAs that act on classical swine fever virus (CSFV) in swine umbilical vein endothelial cells (SUVECs). The 3'-and 5'-untranslated regions ( UTR) of CSFV were clon... The aim of the study was to identify the microRNAs that act on classical swine fever virus (CSFV) in swine umbilical vein endothelial cells (SUVECs). The 3'-and 5'-untranslated regions ( UTR) of CSFV were cloned and then inserted into psiCHECH TM-2 plasmids carrying Firefly and Renilla luciferases reporter genes; microRNAs that acted with CSFV were predicted by bio-informatics analysis; then the recombinant plasmids and inhibitors of the predicted microRNA were co-transfected into SUVECs. The activities of Firefly and Renilla luciferases were detected by luminometry. The PCR products of the CSFV 5'-UTR (373 bp) and 3'-UTR (252 bp) were detected by electrophoresis on 1% agarose gels. Four microRNAs (ssc-miR-let7c, ssc-miR-106a, ssc-miR-18, ssc-miR-139) were screened out and evaluated. The CSFV 3'-UTR is the important target site of microRNA in SUVECs. The four microRNAs mentioned above had different inhibitory effects on the CSFV 3'-UTR, of which the ssc-miR-18 played the most important role. 展开更多
关键词 脐静脉内皮细胞 猪瘟病毒 血管内皮细胞 MICRORNA 荧光素酶报告基因 小分子RNA 重组质粒 电泳检测
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2株猪源产肠毒素型大肠杆菌的分离鉴定及毒力因子和耐药性分析
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作者 马欣 刘博 +6 位作者 魏津 张效斌 朱良全 李霆 王琦 王雨雨 姚文生 《中国兽药杂志》 2026年第2期42-53,共12页
从湖南某猪场断奶后腹泻仔猪粪便中分离到2株疑似大肠杆菌,命名为H株和J株。为鉴定其致病性大肠杆菌类型,本研究将H株和J株分别接种伊红美蓝琼脂平板鉴别培养基和大肠杆菌IMViC生化鉴定试剂,并采取分子鉴定技术对其进行微生物种属鉴定;... 从湖南某猪场断奶后腹泻仔猪粪便中分离到2株疑似大肠杆菌,命名为H株和J株。为鉴定其致病性大肠杆菌类型,本研究将H株和J株分别接种伊红美蓝琼脂平板鉴别培养基和大肠杆菌IMViC生化鉴定试剂,并采取分子鉴定技术对其进行微生物种属鉴定;同时采取PCR扩增技术分析其携带大肠杆菌的肠毒素、黏附素以及α-溶血素、志贺样毒素2型变异体(Stx2e)、耶尔森氏菌毒力岛(HPI)等毒力因子情况。为了解H株和J株的耐药性,本研究使用微量肉汤稀释法对其进行最低抑菌浓度(MIC)测定,并参照CLSI M100-S33抗微生物药物敏感性试验执行标准进行耐药性分析。结果显示,H株、J株的伊红美蓝琼脂平板培养特性、大肠杆菌IMViC生化鉴定试验结果、分子鉴定结果均符合大肠杆菌典型特征;毒力因子分析结果为:H株为LT^(+)+STa^(+)+STb^(+)+EAST-1^(+)+F18^(+)+Stx2e^(+)+α-溶血素+,J株为STa^(+)+STb^(+)+HPI^(+);耐药性分析结果为:H、J株均对氨苄西林、四环素、氟苯尼考、磺胺异噁唑、复方新诺明耐药和对阿莫西林/克拉维酸、庆大霉素、头孢他啶、恩诺沙星、美罗培南敏感;不同的是H株对大观霉素、粘杆菌素耐药和头孢噻呋中介,而J株对大观霉素、头孢噻呋敏感和粘杆菌素中介。本研究上述结果表明H株、J株均为产肠毒素型大肠杆菌(ETEC),但携带的毒力因子各不相同,且均存在多重耐药,与浙江、河北、江西、福建和广西的猪源ETEC耐药情况的比较分析表明养殖环节大肠杆菌耐药性具有复杂性。本研究既为大肠杆菌毒力因子研究和猪场防控大肠杆菌感染引起的仔猪断奶后腹泻病(PWD)提供了参考,也为如何减少兽用抗菌药的使用提供了新思路:兽用抗菌药+疫苗+益生菌+噬菌体+益生元的综合防控手段。 展开更多
关键词 猪源 产肠毒素型大肠杆菌 分离鉴定 毒力因子 耐药性
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猪玉米净能值研究进展
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作者 塔胜男 李柏浩 +6 位作者 班志彬 陈思淼 赵晓东 梁浩 张琦玉 李立佳 闫晓刚 《中国饲料》 北大核心 2026年第1期26-29,共4页
玉米是我国重要的粮食作物,同时也是主要的饲料原料。在我国人畜争粮矛盾日益加剧的背景下,如何提高饲料原料的利用效率已经成为亟待解决的问题。通过运用净能体系对玉米净能值进行评价,能够充分合理利用玉米资源,保障粮食安全。本文主... 玉米是我国重要的粮食作物,同时也是主要的饲料原料。在我国人畜争粮矛盾日益加剧的背景下,如何提高饲料原料的利用效率已经成为亟待解决的问题。通过运用净能体系对玉米净能值进行评价,能够充分合理利用玉米资源,保障粮食安全。本文主要综述了净能体系的优势、测定方法以及猪玉米净能值测定的研究进展,以期为猪玉米净能体系研究提供参考。 展开更多
关键词 玉米 净能 测定方法
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CO_(3)^(2-)与Ca^(2+)对磷酸铵镁结晶法去除模拟猪场厌氧消化液中磷的影响机制
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作者 贾博涵 孙晨 +2 位作者 谢晓琳 钱锋 宋永会 《环境工程技术学报》 北大核心 2026年第2期507-515,共9页
为提高养殖废水厌氧消化液中磷的回收效率与产物资源化价值,以模拟猪场厌氧消化液为对象,系统探究了碳酸根(CO_(3)^(2-))与钙离子(Ca^(2+))对磷酸铵镁(MAP)结晶法除磷过程的影响机制。通过批次沉淀实验,结合Visual MINTEQ 3.0热力学模型... 为提高养殖废水厌氧消化液中磷的回收效率与产物资源化价值,以模拟猪场厌氧消化液为对象,系统探究了碳酸根(CO_(3)^(2-))与钙离子(Ca^(2+))对磷酸铵镁(MAP)结晶法除磷过程的影响机制。通过批次沉淀实验,结合Visual MINTEQ 3.0热力学模型与X射线衍射分析,揭示了不同pH与镁、氮、磷物质的量比[n(Mg)∶n(N)∶n(P)]条件下磷的去除规律与产物特征。结果表明:在pH=9.5、n(Mg)∶n(P)=1.4∶1时,磷去除率超过96%,继续提高pH或n(Mg)∶n(P)对去除率的提升作用不显著。CO_(3)^(2-)单独存在对磷去除率(>95%)及MAP晶体纯度影响较小;而Ca^(2+)单独存在则显著改变沉淀产物组成,当n(Mg^(2+))∶n(Ca^(2+))=2∶1时,MAP纯度约为66.7%,随着Ca^(2+)浓度升高,产物逐渐转为无定形磷酸钙主导。CO_(3)^(2-)与Ca^(2+)共存时的影响与Ca^(2+)单独存在时基本一致,高Ca^(2+)/CO_(3)^(2-)背景下虽能维持高磷去除率(≥95%),但产物纯度显著下降。在实际应用中,应根据离子浓度比精确控制镁与磷的物质的量比,以平衡磷的去除率及回收产物的资源价值。 展开更多
关键词 猪场厌氧消化液 磷酸铵镁(MAP) 磷去除 竞争沉淀 磷回收 MINTEQ模型
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基于母猪VBP1,CpG_280和CpG_1360靶标DNA甲基化的妊娠检测技术建立
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作者 顾兰英 李世义 +2 位作者 张秦川 孙延鸣 张彦兵 《畜牧兽医学报》 北大核心 2026年第3期1458-1469,共12页
旨在筛选母猪妊娠相关差异甲基化基因,建立基于甲基化荧光探针早孕检测技术。本研究重点关注基因启动子和CpG岛差异甲基化区域,筛选潜在的猪妊娠靶标。首先利用甲基化特异性PCR(methylation-specific PCR,MSP)对基因差异甲基化区域进行... 旨在筛选母猪妊娠相关差异甲基化基因,建立基于甲基化荧光探针早孕检测技术。本研究重点关注基因启动子和CpG岛差异甲基化区域,筛选潜在的猪妊娠靶标。首先利用甲基化特异性PCR(methylation-specific PCR,MSP)对基因差异甲基化区域进行初步验证,进一步设计靶标荧光探针,建立基于甲基化荧光探针(quantitative methylation-specific PCR,qMSP)早孕检测技术方法并进行验证。试验数据显示,发现相比于空怀母猪,妊娠25~30天母猪血液中VBP1和CpG_280呈现高甲基化状态,而CpG_1360表现为低甲基化,表明这些位点可能参与妊娠调控。以猪β-actin为内参探针,设计VBP1、CpG_280和CpG_1360甲基化探针,建立qMSP检测方法并初步应用。对靶标基因甲基化探针检测技术与B超检测符合率进行分析,VBP1、CpG_280和CpG_1360的受试者工作特征曲线(receiver operating characteristic curve,ROC)下的面积(area under curve,AUC)分别为0.7138、0.6080和0.7466,均大于0.5,基于VBP1、CpG_280和CpG_1360的qMSP检测技术符合率分别为85.0%、78.75%和85.0%,AUC值大于0.5说明该检测技术具有诊断价值。本研究证实在母猪妊娠早期其血液中VBP1和CpG_280为高甲基化而CpG_1360为低甲基化,基于靶标基因的甲基化荧光探针检测技术可作为一种早期诊断猪妊娠的新方法。 展开更多
关键词 DNA甲基化 妊娠诊断 荧光探针 早孕
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