奶牛特异性转移因子的制备及其免疫活性检测

通过冷冻透析法制备奶牛隐性乳房炎特异性转移因子,并对其免疫活性进行检测。方法为采用匀浆、冻融、透析被免疫猪的脾脏,制备特异性转移因子,运用理化性质检验、安全试验、无菌试验检验所制备转移因子的纯度及安全性,并运用小鼠巨噬细胞吞噬功能试验及E玫瑰花环试验对特异性转移因子免疫活性进行检测。试验结果为:所制备的奶牛隐性乳房炎特异性转移因子多肽含量为1.3 p g/mL,核酸含量为24μg/mL,OD_(260 nm)/OD_(280 nm)值为2.08,且安全、无菌。巨噬细胞吞噬功能试验及E玫瑰花环形成试验表明转移因子可明显增加巨噬细胞吞噬能力并能增加E玫瑰花环的形成数。因此,本品可作为奶牛隐性乳房炎的治疗药物,亦可作为细胞免疫增强剂使用。     

新必妥对仔猪体液免疫效果的影响

本试验选取21日龄的健康仔猪12头，随机分为3组，分别肌肉注射猪瘟兔化弱毒疫苗1头份（对照组）、猪瘟兔化弱毒疫苗1头份＋猪脾转移因子0．15mL／kg体重（试验1组）和猪瘟兔化弱毒疫苗1头份＋猪脾转移因子0．25mL／kg体重（试验2组），分别于21、60日龄各免疫1次．以研究猪脾转移因子对仔猪猪瘟兔化弱毒疫苗免疫效果的影响。结果表明．首免后14～35d试验组的免疫效果均显著优于对照组，二免后5～15d试验2组显著优于对照组。另外，选取35日龄的健康仔猪12头，随机分为3组，分别肌肉注射猪蓝耳病弱毒疫苗1头份（对照组）、猪蓝耳病弱毒疫苗1头份＋猪脾转移因子0．15mL／kg体重（试验1组）和猪蓝耳病弱毒疫苗1头份＋猪脾转移因子0．25mL／kg体重（试验2组），以研究猪脾转移因子对仔猪蓝耳病弱毒疫苗免疫效果的影响。结果表明，免疫后14～35d试验组的免疫效果均显著优于对照组，42～60d试验2组显著优于对照组。表明猪脾转移因子对仔猪体液免疫有一定的增强作用。     

猪圆环病毒2型感染对猪瘟疫苗免疫的影响

猪瘟(CSF)被世界动物卫生组织(OIE)列为必须报告的A类传染病之一,是我国目前强制免疫的主要传染病之一.做好猪瘟免疫是制定猪场免疫程序的基础.近年来,猪瘟疫苗免疫效果不佳甚至免疫失败的现象时有发生,除了猪瘟疫苗质量、免疫程序不当和猪瘟野毒持续感染母猪将其野毒传播给仔猪造成免疫失败等因素外,其他猪病病原感染对猪瘟疫苗的免疫效力也构成影响[1-4].由猪细小病毒(PPV)、猪伪狂犬病病毒(PRV)和猪繁殖与呼吸综合征病毒(PRRSV)单独或混合感染对猪瘟弱毒疫苗免疫效力的影响已有报道[5-8],而由猪圆环病毒(PCV)感染是否猪瘟免疫有影响还鲜有报道.     

转移因子与重组转移因子及其在疾病防治中的应用

转移因子是来自于免疫淋巴细胞的一类可透析小分子多肽,它能够将致敏淋巴细胞的免疫信息传递给未致敏的受体淋巴细胞。转移因子具有特异性和非特异性免疫活性,特异性转移因子具有转移特异性细胞免疫的能力,非特异性转移因子具有促进淋巴细胞活化增殖,增强其免疫活性,并可诱导靶细胞产生大量的细胞因子。重组转移因子是利用基因工程技术将编码具有免疫增强活性的转移因子基因,进行克隆和体外表达,表达产物经纯化制备而成,试验表明其具有增强细胞免疫活性的作用。转移因子和重组转移因子是一种具有免疫增强活性的新型高效安全的生物药品,在临床上用于疾病的防治,具有广阔的应用前景。     

猪瘟活疫苗(种毒)鉴别检验阳性血清制备

为制备猪瘟活疫苗(种毒)鉴别检验阳性血清,本实验将4头非免疫健康猪进行猪瘟活疫苗基础免疫和猪瘟石门强毒株反复强化免疫后,采血分离血清,过滤、分装、冻干、熔封,获得4批产品,并进行了检验和验证。结果显示:物理性状、无菌检验、安全检验、均一性检验、支原体检验、真空度测定均合格;剩余水分均小于4%;特异性检验表明,口蹄疫病毒、猪伪狂犬病病毒、猪繁殖与呼吸综合征病毒、猪细小病毒、牛病毒性腹泻/粘膜病病毒抗体均为阴性;残余猪瘟病毒免疫荧光和套式RT-PCR核酸检测结果均为阴性;兔体中和效价分别为1∶800、1∶800、1∶800和1∶1 600;对猪瘟兔化弱毒种毒和5种猪瘟活疫苗的验证结果均良好。该4批血清可以用于猪瘟活疫苗(种毒)鉴别检验。     

猪几种传染病的药物治疗与预防

1 猪瘟 发生猪瘟时,淘汰病猪并消毒,有条件时可用抗猪瘟血清肌肉注射,每千克体重1毫升,每天1次,连用2次.猪用转移因子加干扰素,分别肌肉注射,或排疫肽加转移因子,分别肌肉注射,每天1次,连用3天,配合对症治疗.未发病的猪只用猪瘟兔化弱毒疫苗紧急注射,每头肌肉注射8～10头份,同时肌肉注射转移因子1毫升,可有效地控制疫情.     

免疫增强剂对猪瘟疫苗免疫效果的影响

为查明免疫增强剂是否能够提高猪瘟疫苗免疫抗体水平,选用猪用转移因子与黄芪多糖注射液直接稀释猪瘟兔化高效细胞弱毒苗的方法进行猪瘟疫苗免疫试验。与对照组相比,猪用转移因子直接稀释猪瘟兔化高效细胞弱毒苗组S/P平均值首免后10 d提高127.27%,差异显著（P＜0.05）;首免后20 d提高156.60%,差异极显著（P＜0.01）;整个试验期提高57.05%。与黄芪多糖注射液直接稀释猪瘟兔化高效细胞弱毒苗组相比,S/P平均值首免后10 d提高131.48%,差异显著（P＜0.05）;首免后20 d提高91.55%,差异显著（P＜0.05）;整个试验期提高31.84%。证明猪用转移因子直接稀释猪瘟疫苗可提高猪瘟疫苗的免疫抗体水平;同时也揭示采用断奶前后首免、60日龄左右二免的猪瘟疫苗免疫程序,仔猪在首免后10～30 d免疫抗体S/P值低,阳性率在60%以下,易感染猪瘟野毒。     

猪瘟特异性转移因子对猪瘟疫苗免疫抗体水平影响的研究

本试验对猪瘟特异性转移因子和猪瘟疫苗联合应用初免和二次免疫后,抗体消长情况进行测定,研究猪瘟特异性转移因子对猪瘟疫苗免疫抗体水平影响规律。结果表明,猪瘟特异性转移因子对提高猪瘟免疫抗体效果极为显著,与使用普通转移因子相比具有明显的效果。     

不同动物种类转移因子对家禽免疫效果影响的对比性研究

为了验证猪、鸡、牛三种转移因子(transfer factor, TF)对家禽免疫效果的影响,试验以鸡新城疫(newcastle disease,ND)血凝抑制试验的抗体效价为判定指标,比较三种转移因子的免疫增强效果。将162只鸡随机分为4组采血并接种鸡新城疫弱毒疫苗,其中猪转移因子组、鸡转移因子组和牛转移因子组(各50只鸡)分别口服猪、鸡和牛转移因子各1 mL,对照组(12只鸡)口服生理盐水1 mL;之后第1,3,5,7,9,11,14,21天采血,用血凝抑制试验检测抗体效价。结果表明:三种转移因子对鸡新城疫弱毒疫苗的免疫效果均有增强作用,其中鸡转移因子组增强作用最明显,猪转移因子组次之,牛转移因子组虽有一定免疫增强作用但效果不如另外两组。说明临床应首选鸡转移因子,其次可选择猪转移因子,牛转移因子需酌情使用。     

犬特异性转移因子的制备及其免疫活性检测

采用冻融-透析法提取了猪脾脏转移因子(TF),并对其理化特性及免疫活性进行了检测。首先,利用环磷酰胺建立小鼠免疫抑制模型,通过MTT法比较犬脾特异性转移因子对免疫抑制小鼠、生理盐水接种小鼠和正常小鼠的淋巴细胞转化的作用。结果表明犬脾特异性转移因子和PHA对免疫抑制小鼠的外周血淋巴细胞转化具有明显的免疫增强作用,与对照组相比差异极显著(P0.01),其中犬脾特异性转移因子免疫增强作用显著高于PHA组(P0.01)。而对生理盐水组和正常小鼠的外周血淋巴细胞转化与免疫抑制小鼠基本一致,犬脾特异性转移因子和PHA对生理盐水组和正常小鼠的淋巴细胞均表现出一定的免疫增强活性,犬脾特异性转移因子免疫增强作用显著高于PHA组(P0.01)。     

Ivermectin: its effect on the immune system of rabbits and rats infested with ectoparasites.

The influence of subcutaneously administered ivermectin on the specific immune response was studied in rabbits infested with mites (Psoroptes cuniculi) and in rats infested with lice (Polyplax spinulosa). A pronounced specific antibody activity and a change in immunoblotting pattern was observed in rabbits after the ivermectin treatment. However, in rats the antibody activity decreased and the profile of specific antibodies, tested by immunoblotting, remained the same as before the treatment. The specific immune response in rabbits artificially immunized with whole-body Psoroptes cuniculi extract was not affected by ivermectin. It was concluded that ivermectin has no direct effect on the immune response of rabbits and rats and that the enhanced immune response in the mite-infested rabbits was caused by the massive release of antigens associated with the synchronous death of the mites.     

Comparisons of nonspecific and specific immunomodulation by oxolinic acid, oxytetracycline and levamisole in salmonids

Oxolinic acid, a promising drug for the treatment of bacterial fish disease agents, was tested for possible immunomodulatory effects on fish. Another antibiotic oxytetracycline, known to be immunosuppressive at higher treatment doses, and levamisole, a known immunostimulator for higher vertebrates, were also compared for causing changes in the nonspecific defense compartment and the specific immune system in rainbow trout. Groups of fish were immunized with Yersinia ruckeri O-antigen bacterin in combination with selected doses of the drugs. The nonspecific defense activity was measured by demonstrating neutrophil metabolic activity by the nitroblue tetrazolium assay, by counting engulfed bacterial cells for a phagocytic index and by counting leukocytes with adherent bacterial cells for the adherence index. The specific immune response was monitored by the passive hemolytic plaque assay demonstrating the numbers of antibody-producing cells. The results showed that oxolinic acid, used at recommended doses for the treatment of bacterial diseases, did not cause immunosuppression in either the nonspecific defense or specific immune system compartments, whereas tetracycline at 10 mg/kg caused reduced activity in both. Fish given levamisole injections before the antigen injection showed a stimulated nonspecific defense but a much reduced specific immune response.     

Bovine herpesvirus-1 infection of cattle: kinetics of antibody formation after intranasal exposure and abortion induced by the virus

The kinetics of antibody formation in Holstein heifers after primary and secondary intranasal inoculation of bovine herpesvirus-1 (BHV-1) and after BHV-1-induced abortion was determined. Sera were fractionated by gel filtration and ion-exchange chromatography. The antibody activity within serum immunoglobulin (Ig) isotypes was assessed, using a plaque-reduction neutralization assay. The primary immune response to BHV-1 infection was characterized by the appearance of IgM and IgG antibodies in serum by postinoculation day (PID) 7. Maximal IgG antibody activity occurred at PID 35 in nonpregnant heifers and at PID 14 in pregnant heifers. Thereafter, IgG antibody activity declined slowly in both groups of heifers. Maximal IgM antibody activity occurred at PID 14 in both groups of heifers and declined rapidly thereafter. The IgG antibody activity during primary immune responses was restricted to the IgG1 subclass. Secondary responses were characterized by anamnestic IgG antibody responses. Antibody activity was present within the IgG1 and IgG2 subclasses during secondary immune responses, but the increase in antibody activity during this period was primarily in the IgG2 subclass. Secondary IgM antibody formation was elicited by abortion induced by the intra-amniotic inoculation of BHV-1, but not by reexposure by the intranasal route. Abortion occurred in 1 heifer 28 days after intranasal BHV-1 inoculation. Abortion in this heifer was not associated with a secondary antibody response. The nature of BHV-1 antigenic exposure in the bovine determined the relative distribution of anti-BHV-1 antibody activity in serum IgM, IgG1, and IgG2. The formation of IgM antibody, with the exception of secondary intranasal exposure, indicated recent BHV-1 antigenic exposure.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro Studies of Group E Streptococci in Swine Leukocytes I. Phagocytic and Bactericidal Properties of Polymorphonuclear Leukocytes from Swine

Serum from both immune and nonimmune ten-week-old swine contained factors which promoted phagocytosis of group E Streptococci (GES). The factors in nonimmune serum, which were heat labile at 70°C for ten minutes, were less efficient than the factors present in immune serum.

Bactericidal activity of the polymorphonuclear (PMN) leukocytes against GES was observed with serum from both immune and nonimmune ten-week-old swine, as well as with serum from normal sows and piglets. However, the bactericidal activity of PMN leukocytes in serum from either normal sows or immune ten-week-old swine was greater than the bactericidal activity of PMN leukocytes in either piglet serum or serum from nonimmune ten-week-old swine. When the serum was either heated to 70°C for ten minutes or treated with 2-mercaptoethanol, bactericidal activity of PMN leukocytes against GES was only observed in the presence of immune serum.

     

Effect of sulphadiazine and trimethoprim on the immune response of rainbow trout (Oncorhynchus mykiss)

A combination of sulphadiazine and trimethoprim at a ratio of 5:1 (SDZ/TMP) was tested for possible immunomodulatory effects. The aim of the study was to follow the immune response after vaccination with simultaneous drug treatment. The fish were immunised with a commercial oil-based divalent (furunculosis/vibriosis) vaccine and were simultaneously given oral drug treatment. The specific immune response was monitored by analysing the levels of specific antibodies with ELISA. As indicators of the nonspecific immune response, the lysozyme activity of serum was measured and the phagocytic activity of circulating leucocytes was monitored by a chemiluminescence assay. Total circulating leucocyte counts and differentials were also monitored. The disease resistance was evaluated by challenge tests at the end of the experiment.The results indicate that SDZ/TMP at a ratio of 5:1 does not interfere negatively with the immune response in rainbow trout after vaccination. A slight stimulation in the antibody response as well as in the chemiluminescence response of circulating granulocytes was recorded in fish treated with the drugs in connection with vaccination. The drugs did not significantly affect the survival after challenge.     

SPF雏鸡感染网状内皮组织增殖病病毒后肿瘤坏死因子的动态变化

采用微量细胞病变法和四甲基偶氮唑盐（ＭＴＴ）法研究了１日龄ＳＰＦ雏鸡感染网状内皮细胞增殖病病毒（ＲＥＶ）后,免疫器官脾脏肿瘤坏死因子（ＴＮＦ）诱生活性动态变化。结果表明,雏鸡感染ＲＥＶ后脾脏细胞ＴＮＦ诱生活性极显著增高（Ｐ〈０．０１）,ＴＮＦ诱生活性明显增高与ＲＥＶ在机体各器官内长期大量繁殖紧密相关,并引起各免疫器官严重的变质性变化,以及由于ＴＮＦ高含量的免疫损伤作用而降低机体各免疫器官的机能。     

Modulation of juvenile brook trout (Salvelinus fontinalis) cellular immune system after Aeromonas salmonicida challenge

In fish, the first line of defence against infectious microorganisms is based on non-specific cellular immune mechanisms (innate immunity). In this study, we measured the non-specific immune parameters (natural cytotoxic cells (NCC) activity, lymphoproliferation, percentage of phagocytosis and phagocytic activity) in brook trout (Salvelinus fontinalis) infected by a virulent strain of Aeromonas salmonicida. Eight days post-infection, the mortality of infected fish reached 70%. A transient immunostimulation of the NCC activity was noticed 24h post-infection, but there was no significant difference at 48 h. Then, infection of brook trout with A. salmonicida induced a biphasic immune response. At 24h post-infection, lymphoproliferation was drastically depressed but returned to control level at 96 h. A slight increase in the percentage of phagocytosis and the phagocytic activity was noticed throughout the experiment. Conversely the cell mortality was significantly higher in infected fish compared to control. The modulation of immunological parameters might reveal important clues on how innate immunity might protect fish from bacterial infections.     

Immunosuppression in bitches with pyometra

The function of the immune system was studied in 34 bitches affected by pyometra. The mean age of the animals was 8.1 +/- 2.8 years. Samples of peripheral blood were examined using a battery of immunological tests. Data obtained from four sets of animals, grouped according to the severity of their disease, were compared. Leucocytosis (neutrophilia and partial monocytosis) and lymphopenia were detected in 62 per cent and 35 per cent of the bitches, respectively. The increase in the number of leucocytes was higher in the severely affected bitches. Inhibition of mitogen-driven lymphocyte proliferation was the characteristic feature of the immunological profile in bitches affected by pyometra, and this corresponded with the impairment of the general state of health. Sera from bitches showing functional inhibition of lymphocytes suppressed the activity of lymphocytes isolated from normal dogs. Sera from bitches affected by pyometra also had higher levels of immunoglobulins, lysozyme and circulating immune complexes. It can be concluded that pyometra is associated with an inflammatory response, but also with a marked suppression of immune system activity.     

肽聚糖对鲫鱼嗜水气单胞菌灭活疫苗免疫增强效果的研究

为了探讨肽聚糖在鱼类抗嗜水气单胞菌（Aeromonas hydrophila，AH）感染中的免疫佐剂作用，用添加A3a肽聚糖的饲料投喂接种F-AH疫苗的彭泽鲫（Carassius auratus var．pengze）；通过测定鲫白细胞吞噬活性、血清和体表黏液溶菌酶活性、抗体效价以及活菌攻毒后的免疫保护率，证明肽聚糖与F-AH疫苗联合使用，鲫体表黏液和血清溶菌酶活性、白细胞吞噬活性、抗体效价以及活菌攻毒后的免疫保护率均显著提高，表明A3a肽聚糖是F-AH疫苗的良好佐剂，能增强鱼类F-AH疫苗免疫应答。     

雏鸡脑软化症免疫组织的氧化和抗氧化特性

用低维生素E（VE）日粮饲喂雏鸡，复制脑软化症动物模型，通过对雏鸡血清和免疫组织中一氧化氮（NO）、丙二醛（MDA）和活性氧（ROS）含量以及黄嘌呤氧化酶（XOD）、谷胱甘肽过氧化物酶（GSH-Px）活性和总抗氧化能力（T-AOC）的检测，探究脑软化症雏鸡免疫组织的氧化与抗氧化特性。研究结果表明，患病雏鸡血清、脾脏和法氏囊中自由基和脂质过氧化物增多，而GSH-Px活性和T-AOC均降低，说明脑软化症雏鸡免疫组织发生氧化应激，抗氧化防御系统机能下降。