临床病猪铁异位沉积的病理组织学观察

采集，临床病猪的病料，采用冰冻切片技术和组织化学染色方法，对猪铁异位沉积的组织学变化进行了显微观察。结果显示，一些病猪的淋巴结、淋巴索和淋巴窦内的淋巴细胞数量显著减少，巨噬细胞的数量大幅度增加；大量吞噬有含铁血黄素颗粒的巨噬细胞聚集在一起，连成一片，致使细胞轮廓不清；它们的趋化运动能力和吞噬处理抗原的机能丧失。证明仔猪过量补充铁制剂，可以引起铁的异位沉积；铁在巨噬细胞内的沉积导致机体的免疫机能下降，使仔猪易患各种传染病。     

Investigations on iron bioavailability of different sources and supply levels in piglets

Two experiments were conducted to investigate the iron bioavailability of different sources and supply levels in piglets. In experiment 1, the influence of feeding an iron deficient basal diet or the basal diet supplemented with 28 ppm Fe either in form of Fe-chelate, crystalline Fe-glycinate, or Fe-sulphate on apparent iron digestibility and on blood parameters was determined. In experiment 2, a dose-response approach was used to determine the effects of supplementing an iron-deficient basal diet with 30, 60, or 90 ppm of Fe either in form of crystalline Fe-glycinate or Fe-sulphate on digestibility of Fe, growth and blood parameters. In experiment 1, comparison of iron sources revealed a tendency (0.05 < p < 0.1) for a higher Fe digestibility from Fe-glycinate (40.9%) compared with Fe-chelate (30.8%) or Fe-sulphate (30.7%). Fe-balance was higher (p < 0.05) for Fe-glycinate compared with Fe-sulphate but iron balance was similar for iron sulphate and iron chelate. Those results, however, were not reflected in data of blood parameters. In experiment 2, iron digestibility, blood parameters and performance were significantly (p < 0.05) influenced by iron supply. Effects of iron source on digestibility of iron were lower than observed in the first experiment.     

饲喂母猪不同类型和水平的有机铁对仔猪生长和贫血状态的影响

本研究旨在评估母猪饲料中添加两种不同水平螯合态的有机铁对仔猪生长性能和改善仔猪贫血效果的研究。选取体重相近、胎次相近、遗传结构一致、健康的二元杂母猪30头,随机分为5组。对照组1组,在基础饲料中添加硫酸亚铁100 g/kg;处理组2、3、4、5分别在基础饲料中添加甘氨酸亚铁80 g/kg和100 g/kg、复合氨基酸螯合铁80 g/kg和100 g/kg。结果表明,母猪饲料中添加有机铁甘氨酸亚铁100 g/kg和复合氨基酸螯合铁80 g/kg可提高21日龄断奶仔猪体重,分别提高12.26%和10.3%;母猪饲料中添加有机微量甘氨酸亚铁和复合氨基酸螯合铁可改善母猪生产时和产后21日龄以及仔猪初生和断奶后体内血液红细胞数量(RBC)、血液平均红细胞的体积(MCV)和血液中红细胞压积(HCT),其中母猪日粮中添加甘氨酸亚铁80 g/kg和复合氨基酸螯合铁100 g/kg组仔猪血液红细胞数量(RBC)在初生和21日龄断奶分别提高6.06%和9.29%;母猪饲料中添加有机微量甘氨酸亚铁和复合氨基酸铁都可提高母猪初乳和常乳中乳铁的含量,其中添加甘氨酸亚铁100 g/kg和复合氨基酸螯合铁100 g/kg组初乳中铁浓度分别提高276%和346%。综上,通过提高母猪乳中有机铁含量可提高初生仔猪体内的铁储存与代谢以达到仔猪抵抗贫血风险的作用。     

Tolerable upper intake level of iron damages the liver of weaned piglets

Iron is one of the essential trace elements, which is often supplemented as an additive to meet the growing needs of toddlers and young animals. Recommended nutrient intake (RNI) and tolerable upper intake levels (UL) are always set when the iron is supplemented. The purpose of this study was to evaluate the subacute (28 days) toxicity of UL iron to weaned piglet liver. Thirty 23-day-old weaned piglets were divided into three groups and, respectively, supplemented with 100, 300 or 3000 (UL) mg/kg iron. UL iron caused significant weight loss in 4th week (p < 0.05). Divalent metal transporter 1(DMT1) decreased significantly, ferroportin 1 and ferritin increased significantly in the liver of UL iron group (p < 0.05). Although there was no significant effect on liver morphology, UL iron significantly increased hepatic iron, reactive oxygen species (ROS), malondialdehyde (MDA) and protein carbonyl (p < 0.05). UL iron significantly reduced glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT) and total anti-oxidation capacity (T-AOC) in the liver (p < 0.05). Nuclear factor erythroid 2-related factor 2 (Nrf2) activated subunits of glutamate cysteine ligase (Gclc) and glutathione S-transferase A1 (Gsta1) upregulation in the UL iron group liver, thereby increasing resistance to oxidative stress. In conclusion, UL iron supplementation altered iron metabolism, generated free radicals, reduced antioxidant enzyme activity and activated Nrf2 signalling pathway in the weaned piglet liver.     

Properties of a Complex of Iron and a Sorbitol-Gluconic Acid Polymer for Treatment of Piglet Anaemia

The properties of a complex of iron and a sorbitol-gluconic acid polymer useful for intramuscular treatment of piglet anaemia have been studied. In rabbits it is well absorbed from muscle and a small percentage (8 %) of the administered iron is excreted in the urine. A satisfactory utilization of the administered iron for haemoglobin synthesis has been shown in rats. A high degree of tolerance in piglets was observed also for the highest test doses, 600 mg iron per kg body weight. The clinical utilization was compared to that of iron-dextran using a total of 209 piglets and a dose of 200 mg iron for each animal. Five of these piglets were lost, but these deaths cannot be correlated to the treatment. Of the remaining animals 102 were treated with ISGP and 102 with iron-dextran, and the anaemia-preventing effect was similar for these 2 iron complexes.     

Establishment of major histocompatibility complex homozygous gnotobiotic miniature swine colony for xenotransplantation

To overcome shortages of human donor organs for organ failure patients, we made a commitment to develop gnotobiotic miniature swine as an alternative organ donor source for xenotransplantation. For this, we have constructed an absolute barrier‐sustained gnotobiotic facility. Pregnant sows of gnotobiotic miniature swine, were procured and germfree piglets were obtained by hysterectomy. These were maintained in germfree isolators for about 4 weeks, deprived of colostrum and were fed sterilized soybean milk. They were associated with di‐flora, anaerobic Lactobacillus sp. and Streptococcus sp. After confirmation of successful associations, gnotobiotic piglets were transferred into the facility aseptically. The piglets are maintained on high‐efficiency particle air‐filtered air in and out; maintaining constant room air pressure of 33 ± 3 mmAq, and sterile water and diet. In 10 sessions of hysterectomy, 18 male and 32 female piglets were obtained of which piglets (M six, F eight) died within 5 days. Among live piglets, piglets (M eight, F 12) were confirmed to be germfree by microbiological monitoring. For research of xenotransplantation, one consistent experimental result was essential. Therefore, major histocompatibility complex class II which related innate immunity, homozygotic gnotobiotic miniature swine was developed. As a result, genotyping revealed 14 individuals to be homozygous for major histocompatibility complex class II (DRB, DQB) as 0301, three individuals were homozygous as 0201 and each of two were homozygous for DQB as 0701 and DRB as 0404, respectively. Genetic modifications and immunological research for ideal alternative organ sources are in progress.     

猪瘟病毒Erns基因的克隆及原核表达

从猪瘟病毒石门株血液样品中提取总RNA,通过R-PCR对Erns基因进行cDNA扩增,获得了696 bp的片段.将该片段克隆于T-easy载体后进行序列分析,确认PCR产物为猪瘟病毒Erns基因,从阳性克隆中提取质粒,经Bam H Ⅰ和Hin d Ⅲ双酶切,回收产物亚克隆到pET-32a表达载体中,提取质粒后转化BL21(DE3)感受态细胞,并筛选出阳性克隆,经IPTG诱导后通过SDS-PAGE检测出Erns基因的表达.     

乳铁蛋白在猪病防治及饲料中的应用

本文概述了乳铁蛋白的分子结构及其生物学功能,从促进铁的吸收、调节免疫、抗菌、抗病毒、抗氧化等生理功能方面说明其在仔猪疾病防治及饲料中的应用前景。     

血红素铁对断奶仔猪生长性能和血液指标的影响

选取120头28日龄断奶仔猪,随机分成4个处理组,即1个对照组和3个试验组,每组3个重复,每个重复10头。对照组饲喂基础日粮,试验1、2、3组分别饲喂基础日粮中血红素铁净添加量为27.5、55.0、85.0 mg/kg的3种试验日粮,研究日粮中添加血红素铁对断奶仔猪的生长性能、血液指标(血红蛋白浓度、免疫球蛋白浓度)的影响。试验正试期13 d。结果表明,血红素铁各添加量组均能够提高仔猪的生长性能,日粮中血红素铁添加量为55 mg/kg时,仔猪平均日增重、饲料转化率较对照组分别提高12.02%和10.27%(P<0.05)。血红素铁对断奶仔猪有良好的促生长作用,也在一定程度上提高了仔猪血液中血红蛋白和IgG浓度。     

猪瘟病毒E2蛋白抗原表位的表达与免疫活性研究

以猪瘟病毒全长基因组质粒为模板,PCR扩增出E2囊膜糖蛋白上A/D抗原区B细胞表位878aa～938aa;PCR扩增片段连接到pET-32a( )载体构建pET-AD原核表达质粒,将阳性质粒转化到BL21大肠埃希菌中诱导表达,以Ni-NTA亲和柱纯化,纯化后的蛋白进行Western blot鉴定。结果表明,重组质粒pET-AD在BL21大肠埃希菌中可以高效表达,表达的蛋白质经纯化后仍然有反应原性。     

低聚木糖在猪生产中的应用

本文就低聚木糖的生物学特性及其在猪生产中的应用作一综述。     

猪苏氨酸营养的研究与应用

苏氨酸通常是猪的第二或第三限制性氨基酸,对其生长具有重要作用。猪日粮中添加合成苏氨酸,可提高其生产性能,节约日粮蛋白质,增加养猪效益。本文综述了苏氨酸的理化性质、生物学功能,并介绍了苏氨酸在猪生产中的应用和猪对其需要量及其影响因素。     

非洲猪瘟病毒VP73结构蛋白表达及鉴定

根据GenBank登录的非洲猪瘟病毒(ASFV) VP73基因序列,人工合成VP73主要抗原表位区基因片段,克隆至pUC57载体中.设计1对引物,PCR扩增获得429 bp的VP73基因片段;将VP73基因片段使用EcoRI和SalI酶切后亚克隆至表达载体pGEX-KG,经PCR、酶切、测序鉴定挑选阳性克隆,转化至表达菌株BL21 (DM3)中,进行体外诱导表达,SDS-PAGE和Western blot分析蛋白表达及其活性.结果显示,成功构建表达重组载体pGEX-KG-VP73,对该重组载体进行诱导,有效表达了41 ku重组蛋白,表达量约占菌体总蛋白的17％,Western blot分析证实该蛋白具有良好反应原性.     

猪瘟病毒E2糖蛋白A1抗原亚区的表达

猪瘟病毒(CSFV)囊膜结构糖蛋白E2(gp55)是诱导机体产生中和抗体及激发保护性免疫应答的主要抗原蛋白。E2存在4个不同抗原区(A-D)，根据中和特性和保守性差异，将A抗原区进一步划分为A1、A2、A3抗原亚区，A1抗原亚区在不同猪瘟病毒分离毒株中高度保守且诱导的单克隆抗体具有中和特性。本采用pRSET C载体，对A抗原区2744nt～2947nt(791aa～858aa)基因片段(EC)进行克隆、表达，并分析表达产物的免疫反应性。研究发现：EC表达产物能与抗猪瘟病毒Alfort毒株E2蛋白的中和性单克隆抗体c2410发生特异性结合，且此结合能被猪瘟病毒抗原或阳性血清阻断。结果表明：A1抗原亚区位于E2蛋白791～858位氨基酸区域。     

IGF-I调控区微卫星座位遗传多态性对松辽黑猪肉质性状的影响

在克隆5’上游微卫星序列的基础上,以40头松辽黑猪为研究材料,分析了该微卫星序列的多态性,同时进行了各基因型与肌内脂肪含量等肉质和胴体性状的关联性分析。结果表明：在5’端侧翼微卫星位点上多态性丰富,共检测到4个等位基因（A、B、C、D）共6种基因型,等位基因A、B、C、D的频率分别为0．1125、0．0875、0．4250和0．3750;杂合度0．6580、多态信息含量0．5944、有效等住基因数2．9240;遗传效应分析发现AA基因型个体具有较高的肌内脂肪含量。     

非洲猪瘟病毒p72蛋白杆状病毒表达系统的构建及其抗原性分析

为研究真核表达非洲猪瘟病毒(ASFV)主要结构蛋白p72,本研究从含ASFV p72基因全序列的重组质粒pGEX-6p-p72中扩增出1 941 bp的p72全长基因,将其插入于pFastBac HTa杆状病毒载体中,构建了重组质粒pFastBac HTa-p72,转化至感受态细胞DH10Bac中,获得重组杆粒rBacmid-p72。再将其转染至sf9昆虫细胞,获得重组杆状病毒。Western blot和夹心法ELISA分析表明ASFV p72基因在昆虫细胞sf9中获得了正确表达,重组p72蛋白可以被特异性抗ASFV血清、p72单克隆抗体识别,表明该蛋白特异性强、活性稳定,具有良好的抗原性。为进一步研究p72蛋白的结构、功能和免疫学特性奠定基础。     

非洲猪瘟病毒入胞过程相关蛋白及分子机制的研究进展

非洲猪瘟病毒(ASFV)是非洲猪瘟(ASF)的病原,可引发家猪和野猪急性、传染性出血死亡,目前尚无有效的疫苗和抗病毒药物。ASFV是囊膜病毒,其囊膜蛋白的结构和功能可能是影响病毒入侵和细胞嗜性的重要因素。病毒入胞是病毒感染细胞的第一步,通常是通过细胞表面特定的分子与病毒蛋白相结合吸附于宿主细胞表面,对ASFV入胞过程的病毒蛋白或宿主因子为靶点或可有效抑制ASFV的复制。本文从ASFV入胞过程中起重要作用的囊膜蛋白出发,对ASFV的入胞分子机制进行综述,为ASFV入胞深入研究以及治疗性药物和疫苗的研发提供参考。     

Shotgun质谱筛选与猪瘟病毒囊膜蛋白Ems相互作用的候选宿主蛋白

猪瘟病毒(CSFV) Ems蛋白是瘟病毒属成员特有的囊膜糖蛋白,在病毒吸附和侵入靶细胞过程中发挥重要作用.本研究利用Ems特异性抗体对CSFV石门强毒株感染的PK-15细胞进行免疫共沉淀(Co-IP),对获得的蛋白复合物进行Shotgun质谱鉴定.结果显示,与阴性对照相比,病毒感染细胞样品中含有199种差异蛋白,其中可能包含与Ems作用的宿主细胞蛋白.同时,实验还对Co-IP蛋白样品进行SDS-PAGE,切取差异条带进行质谱分析以进一步验证Shotgun技术的蛋白筛选结果.通过Co-IP和Shotgun质谱分析,本研究筛选出一些可能在CSFV生命周期中与Ems互作用的宿主蛋白,为进一步研究CSFV在宿主细胞中的感染和复制的分子机制奠定基础.     

猪转移因子对犬外周血中性粒细胞数量和杀菌活性的影响

无菌采犬外周血,肝素抗凝,制备不同浓度的粒细胞悬液,分别与不同浓度的大肠杆菌悬液共同培养,采用MTT法测定粒细胞的杀菌效果,确定粒细胞体外培养时杀伤细菌的最优化条件;把不同浓度的猪转移因子(Swine transfer factor,sTF)加入注射sTF前后不同时期分离的犬外周血中粒细胞与E.coli的培养系统中,测定粒细胞的杀菌能力。结果表明,当粒细胞浓度为1.3×106个/mL,E.coli浓度为6×105个/mL时,试验最能反映粒细胞的杀菌效果。当sTF浓度为1.56mg/mL时,对PMN杀菌活性的影响最大;在犬注射sTF后的第2d,中性粒细胞单独杀菌活性最好,并且此时外周血中性粒细胞的数量达到峰值。本研究结果表明,sTF不仅能够增强犬中性粒细胞的杀菌活性,还能增加犬外周血中性粒细胞的数量。     

规模猪场防控非洲猪瘟的生物安全措施

猪场的生物安全措施对该场的疫病预防和控制起到了非常重要的作用。文章介绍了在非洲猪瘟背景下,猪场需要在人员生物安全意识、猪群健康、猪场入口处、猪场出口处、猪场车辆管理等方面需要采取的一系列生物安全措施。供大家参考,以期提升猪场的生物安全管理水平,提高猪场疫病预防控制能力。