旋毛虫肌幼虫排泄／分泌物抗原的研究

为了研究在特定条件下经体外培养的肌旋毛虫幼虫排泄／分泌物（ES）抗原的特性,通过体外培养将所收集到的ES抗原经聚丙烯酰胺凝胶电泳（SDS—PAGE）、琼脂扩散试验进行分析。结果经SDS—PAGE分析出现了比较明显的2条蛋白带,其分子量为45ku、49ku;经琼脂扩散试验ES抗原与犬旋毛虫阳性血清出现了明显清晰的沉淀线,而与犬蛔虫阳性血清、犬绦虫阳性血清之间未出现沉淀线,ES抗原具有较强的特异性。表明研究结果为开展ES抗原的分子生物学的研究、分子生物学基因工程苗的研制打下了良好的基础。     

应用酶联免疫吸附试验快速诊断猪囊虫、旋毛虫的研究

猪囊虫、旋毛虫病严重威胁着人体健康,传统的检验方法是屠宰后剖检、压片镜检,检出病猪销毁处理,造成了很大积极损失.应用酶联免疫吸附试验活体检验猪囊虫、旋毛虫病,对病猪进行有效治疗,减少经济损失.     

犬旋毛虫肌幼虫体外培养排泄分泌物抗原的分析

自1835年由伦敦的J．Paget和R．Owen首次发现旋毛虫,旋毛虫病已传遍世界各地。在我国已在26个省、市、区有该病的发生或流行的报道。在东北地区尤以犬的感染率最高,这几年随着肉用犬养殖业的发展,食用狗肉饮食文化的发展,犬旋毛虫病的流行范围正进一步扩大,发病率逐渐升高,更加严重威胁着人类健康,也给畜牧业及食品工业带来重大的经济损失。     

旋毛虫排泄/分泌抗原研究进展

旋毛虫病由于其对人、畜危害严重历来被人们所重视 ,其免疫防制是当今国内外学者研究的重要方向。通过对旋毛虫几种抗原的对比研究 ,排泄 /分泌抗原效果最好 ,它具有双重的免疫学功能 ,成为众多学者研究的热点 ,人们对排泄 /分泌抗原成分进行了大量研究。作为蛋白质其主要成分是三种糖蛋白 (分子量分别为4.5万 ,4.9万 ,5.3万 ) ,他们可以发生免疫学交差反应。目前人们已经获得了编码这三种蛋白的基因序列 ,并通过分子生物学技术对其基因进行克隆和表达 ,将表达蛋白用于旋毛虫病的防制。本文介绍了旋毛虫排泄 /分泌抗原的组成成分与功能 ,以及检测抗原和免疫原方面研究状况。     

旋毛虫S_3抗原的免疫学特性及其在诊断中的应用

利用放射免疫的方法,对旋毛虫S_3抗原的特异性与敏感性进行了研究,并对实际样品进行了测定.实验结果表明,S_3抗原可检测抗血清的最低滴度为5×10~(-5),与弓形虫、隐孢子虫及锥虫无交叉反应.与猪囊虫、肉孢子虫的交叉反应率分别为10%和5%.对实际阳性样品可全部检出,阴性样品假阳性率为2.5%,对实验大鼠在72h后可全部测出循环抗原.     

单克隆抗体在旋毛虫抗原研究中的应用

扼要叙述了对旋毛虫抗原的提纯与分析、来源与定位的研究概况以及单克隆抗体技术在旋毛虫抗原研究中的应用与展望。     

旋毛虫成虫排泄分泌物对中性粒细胞功能的影响

利用激光共聚焦显微镜和实时荧光定量PCR等技术,探究旋毛虫成虫排泄分泌物(核酸酶抑制剂ATA阻断DNase酶活性条件下)对中性粒细胞胞外诱捕网形成、活性氧生成和细胞因子产生的影响。结果显示,旋毛虫成虫排泄分泌物能够抑制中性粒细胞胞外诱捕网的形成,减少活性氧生成,促进细胞因子IL-1β、IL-10和TNF-α的表达,降低IL-4的表达,对IFN-γ表达水平无明显影响。结果表明,旋毛虫排泄分泌物能够影响中性粒细胞的功能,为进一步研究旋毛虫免疫逃避机制等奠定良好基础。     

不同发育时期旋毛虫脂肪酸组成的研究

为探讨旋毛虫生长发育过程中脂肪酸组分的变化及外界环境对其影响,本研究采用贝尔曼氏装置分别收集旋毛虫肌幼虫和成虫,同时,将少量骨骼肌和肠黏膜进行酯化处理,通过气相色谱质谱联用技术进行分析。结果显示:肌幼虫和成虫的脂肪酸组分涵盖了C12～C22的22种,主要为16、18和20碳脂肪酸,但旋毛虫肌幼虫与成虫的各脂肪酸相对含量有明显差异(p0.05)。分别对旋毛虫肌幼虫和成虫及其各自寄生部位骨骼肌和肠黏膜中的脂肪酸成份进行比较分析,结果显示:各脂肪酸组成相似,但在相对含量上却有显著差异(p0.05)。表明不同发育时期旋毛虫的脂肪酸组成在种类和相对含量上均存在明显差异,这可能与其所处的不同宿主环境和不同发育时期虫体的特殊生理结构有关。     

旋毛虫肌幼虫可溶性抗原、排泄分泌抗原的电泳分析

本文报道了应用十二烷基硫酸钠一聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ），聚丙烯酰胺凝胶等电聚焦（ＩＥＦ）电泳及二维电泳（ＩＥＦ／ＳＤＳ－ＰＡＧＥ）对旋毛虫肌动虫排泄分泌（ＥＳ）抗原和肌幼虫可溶性抗原的分析结果。肌幼虫ＥＳ抗原经ＳＤＳ－ＰＡＧＥ后用考马斯亮蓝染色蛋白质，结果显示１６条蛋白带，分子量范围２１～８０ＫＤ，其中主带９条。ＩＥＦ电泳后分别用ＰＡＳ染多糖、考马斯亮蓝Ｒ－２５０染蛋白质、Ｎｉｌｅ＇ｓ蓝染脂、醋酸ａ－萘酯／坚固蓝染酪酶同工酶，结果肌幼虫ＥＳ抗原分别显示１６，２６、７及０条带；肌幼虫可溶性抗原分别显示２１、３８、４及１１条带。二维电泳后用考马斯亮蓝Ｇ－２５０染色多肤斑点，结果ＥＳ抗原显示多肽斑点６１个；肌幼虫可溶抗原显示１２２个多肽斑点。     

旋毛形线虫分类的研究进展

概述了旋毛形线虫属种分类研究的现状及虫体杂交试验、同工酶酶谱分析、分子生物学及分子遗传学试验等旋毛虫分类方法的研究进展,指出目前国际上已将毛形属分为8个隔离种（即T．spiralis,T1;T．nativa,T2;T．britovi,T3;T．pseudospiralis ,T4;T．murrelli,T5;T．nelsoni,T7;T．papuae,T10：Lzimbabwensis,T11）和3个分类地位尚未确定的基因型（即T6、T8和T9）。     

旋毛虫致小鼠心肌损伤的试验

本研究旨在对重度感染旋毛虫后小鼠心肌损伤的研究。通过对小鼠接种旋毛虫,1 000条/只,分别于不同时间对血清中心肌损伤标志物H-FABP、CK-MB和cTnT进行动态检测,以及心肌组织病理形态学变化进行观察。结果显示,H-FABP比CK-MB、cTnT敏感性高(P<0.05),出现及达到高峰的时间最早;感染旋毛虫后19d～24d,心肌组织损伤最为严重。H-FABP可用于早期旋毛虫病患者心肌损伤的诊断,具有较高的敏感性和特异性。     

感染旋毛虫小鼠心肌能量代谢变化的检测

本试验旨在对感染旋毛虫后小鼠心肌能量代谢变化进行研究。将小鼠接种旋毛虫1 000条/只后,分别检测小鼠心肌血管紧张素Ⅱ(AngⅡ)、过氧化物酶体增殖物激活受体(PPARs)和小鼠组织胰岛素。结果显示,旋毛虫感染小鼠后,AngII、PPARα及胰岛素的含量检测结果均呈现相似趋势,至19²4 d出现峰值,随后下降。试验结果为旋毛虫病的早期诊断、治疗提供基础数据。     

A search for infections of Trichinella spiralis in New Zealand pigs

Abstract

Extract

Following the occurrence of one case of human trichinosis in New Zealand in 1931 (Lynch, 1932 Lynch, P. P. 1932. N.Z. med. J., 31: 216–216.  [Google Scholar]), a search was made for the larvae of Trichinella spiralis in the diaphragms of pigs. The examinations were made by pressing out pieces of muscle between two glass slides, and examining them under the microscope using a low-power objective. Samples from nearly 20,000 pigs were examined without any infections being found (N.Z. Dept. Agric, 1932 N.Z. Dept. Agric. 1932. Annual Report for 1931–32, : 57–57.  [Google Scholar]).     

Comparative assessment of a double antibody enzyme immunoassay test kit and a triple antibody enzyme immunoassay for the diagnosis of Trichinella spiralis spiralis and Trichinella spiralis nativa infections in swine.

Enzyme immunoassays using the triple antibody enzyme linked immunosorbent assay (ELISA) with both Trichinella spiralis spiralis and T. spiralis nativa excretory-secretory (ES) antigens and a commercial Trichinella spiralis enzyme immunoassay test kit were carried out on sera from pigs that were infected with light, moderate and high doses of infective T. spiralis spiralis and T. spiralis nativa respectively. Seroconversion occurred in all pigs given infective Trichinella larvae although no trichinae were recovered from pigs given T. spiralis nativa larvae and examined between days 92 and 99 postinfection by pepsin digestion. Anti-Trichinella antibodies were detected in pigs infected with T. spiralis spiralis and T. spiralis nativa by ELISA using either the homologous or heterologous ES antigen. The commercial Trichinella spiralis enzyme immunoassay test kit also detected anti-Trichinella antibodies in both the T. spiralis spiralis and T. spiralis nativa infected pigs. The commercial test kit did not appear to be as sensitive as the triple antibody ELISA since it usually took two to three days longer for seroconversion to be detected by the former procedure. Finally seroconversion occurred more rapidly in swine infected with T. spiralis spiralis than with pigs receiving comparable doses of T. spiralis nativa.     

Evaluation of techniques for the recovery of live intestinal Trichinella spiralis worms from experimentally infected foxes

Previously described methods for the recovery of intestinal Trichinella worms from rodents are not feasible when applied in larger experimental animals such as foxes. In this study,worm recovery by standard technique of simple incubation of the intestine in saline was compared to embedment of the intestine in an agar gel. The small intestines of Trichinella spiralis infected foxes (4-5 days post inoculation) were slit lengthwise and the two corresponding halves were processed with one of the two incubation methods.Worms were recovered from all samples, and the total worm recovery ranged from 0.2-4.4% of the infection dose. The samples from the standard incubation were very unclear and time consuming to count compared with samples from the agar gel embedment,in which the intestinal debris were kept inside the agar. As the agar gel technique generally yielded higher numbers of worms than the corresponding standard incubation sample, it is with some optimisation, recommended for recovery of intestinal Trichinella worms from foxes.     

Latex agglutination test for detecting Trichinella spiralis infections in pigs using muscle extract

The latex agglutination (LA) test, using muscle-juice samples of pigs experimentally infected with Trichinella spiralis and slaughtered 95 days post-infection (p.i.), gave visible results in 3 min; even in a pig receiving an infection dose as low as 10 larvae. The test appeared reliable and easy to perform without the need for special equipment or sample treatments which are necessary for ante-mortem diagnostic methods. The muscle-juice sample could be obtained by compressing the muscle pieces with the fingers at any time post-mortem and was used undiluted. The results of the LA test using serum or muscle-juice samples correlated with those of the enzyme-linked immunosorbent assay (ELISA). Positive results in the LA test and ELISA appeared 27 days p.i. with the use of sera from the pigs infected with greater than or equal to 600 larvae and 56 days p.i. with the serum of a pig infected with 10 larvae. The complement-fixing antibodies were detected in the sera using complement ELISA 86 days p.i. This assay was negative when muscle-juice samples were used.     

肌旋毛虫免疫亲和层析抗原的制备及其特性

以Sepharose 4B为载体,兔抗旋毛虫多克隆抗体为配体,纯化了旋毛虫的S_3抗原,得到亲和层析抗原.本实验结果表明,存在于S_3抗原中的主要抗原成分,同样存在于亲和层析抗原中,亲和层析抗原的分子量范围在103000～40000 u之间,等电点范围在pH4.7～8.8之间.亲和层析抗原经转印后,在硝酸纤维素膜上至少有7条显色带,其中分子量为48000 u,50000u,58000u和87000u的4种蛋白的抗原性较强,而以分子量为48000u的蛋白抗原性最强.     

母猪不同生长阶段饲养管理技术

在实际生产中根据母猪的不同生产阶段,分为后备期、间情期、妊娠期、临产期、哺乳期和空怀期等几个阶段。为便于饲养管理和提高养殖效益,应按照各个时期的生理特点和对营养成分的需求采取不同的饲喂方式和配制全价的饲料,满足母猪对营养物质的需求,能保持适宜的体况和膘情,适时发情和配种,并生产出更多更优的仔猪,提高母猪繁殖力和养殖效益。该文分别叙述不同阶段母猪的饲养管理要点。