镰形扇头蜱重组肌钙蛋白Ⅰ抗蜱免疫保护效果的初步分析

为了评估镰形扇头蜱肌钙蛋白Ⅰ(TroponinⅠ,TnI)作为候选疫苗分子的潜力,应用肌钙蛋白Ⅰ重组蛋白进行了抗蜱免疫实验。将重组蛋白按常规方法免疫新西兰兔,三次免疫后分别用若蜱、成蜱进行攻虫实验,观察其免疫保护效果。结果发现蜱的吸血行为受到一定的影响,与对照相比,若蜱和成蜱吸附率、饱血率、饱血体重和死亡率有显著差异,而在饱血时间上差异不显著,说明TnI基因的重组表达产物具有一定的免疫保护作用。     

镰形扇头蜱-新基因的分离、克隆、表达和抗蜱免疫保护作用

目的分离、克隆镰形扇头蜱一新基因。方法本实验从镰形扇头蜱半饱血雌蜱唾液腺cDNA文库的100个EST（Expressed Sequence Tag）序列中选取一个带有polyA尾的序列，经5’RACE（Rapid Amplification of cDNA Ends）得到此序列的全长基因，命名为RhHp2，基因全长1002bp，开放阅读框（ORF）从31bp至879bp，共848bp，编码282个氨基酸。经序列比较，RhHp2基因与其它序列在核苷酸水平上基本无同源性，但在氨基酸水平上与其它物种同源性最高可达50．81％。RhHp2基因连接于PET32a（＋）载体后克隆于表达菌B121，1mM IPTG诱导表达，产生分子量约为55kD的融合重组蛋白，并以不溶性包涵体形式存在。经纯化和体外复性的重组蛋白三次免疫新西兰兔后，将镰形扇头蜱未吸血成蜱和若蜱接种于此免疫兔耳，并对接种48h后的蜱上体数、蜱的饱血数量、饱血时间、饱血重量等进行记录。结果发现蜱的吸血行为受到一定影响，若蜱在48h的减虫率（上体数减少）为58％，在整个吸血过程中的死亡数增加，成蜱的饱血体重显著降低。结论此结果表明RhHp2基因的重组表达产物具有一定的免疫保护作用。     

镰形扇头蜱免疫球蛋白结合蛋白的表达及免疫保护分析

为在大肠杆菌中高效表达镰形扇头蜱免疫球蛋白结合蛋白(immunoglobulin binding protein,IGBP),并对表达产物进行免疫保护效果测定,将IGBP基因克隆到pGEX-4T-1载体,转化到感受态细胞BL21,在IPTG诱导下进行表达;表达产物3次免疫家兔,剂量为500μg/只。3次免疫后进行攻蜱试验,观察免疫保护效果。结果发现在IPTG的诱导下,重组质粒pGEX-4T-1-IGBP在大肠杆菌中获得高效表达,产生GST-IGBP融合蛋白,用融和蛋白免疫家兔后,试验组与对照组成蜱饱血重量和死亡率差异显著(P<0.05)。表明重组质粒pGEX-4T-1-IGBP能在大肠杆菌中高效表达,且表达蛋白能够对家兔产生一定程度的抗蜱保护性免疫力。     

家兔抗长角血蜱免疫模型的建立及对长角血蜱成蜱生长发育的影响

每隔2周用长角血蜱成蜱定量感染家兔,成功构建了具有不同抵抗力的家兔免疫模型。ELISA检测结果表明,家兔血清中的抗体效价从初次叮咬后第3周开始呈阳性,随着叮咬次数的增加抗体水平逐渐上升,第11周时达到高峰。具有一定抵抗力的家兔对长角血蜱成蜱的吸血和生长发育影响显著,其抗体水平与长角血蜱成蜱在兔体的吸血周期、死亡率成正相关,与雌蜱的饱血脱落率、饱血体重成负相关。结果进一步证明,蜱的唾液腺是其主要的免疫器官之一。家兔抗体水平的持续期足以使长角血蜱完成一个世代,可以满足试验需求,是抗蜱免疫研究理想的阳性对照模型。同时,血清中特异性抗体的效价也可以作为判断动物对长角血蜱抵抗力强弱的重要指标之一。     

青海血蜱不同发育阶段蛋白分析

考虑到不同发育阶段蛋白多肽及抗原特性方面的差异,本实验利用SDS-PAGE和Westernblotting对青海血蜱的卵、幼蜱、若蜱、成蜱的蛋白进行了分析。SDS-PAGE结果显示,卵出现了7条主带,幼蜱出现了16条主带,若蜱出现了14条主带,成蜱出现了12条主带。它们的蛋白多肽在数量、含量和分子量上存在差异,共同带只有4条,但相邻发育阶段之间的共同带较多。用感染羊血清和免疫羊血清作免疫印迹时,感染羊血清在分子量5.6万处有明显的反应带,免疫羊血清在分子量9.7万处有明显的反应带。     

镰形扇头蜱肌钙蛋白I基因的克隆及其分布

本试验从镰形扇头蜱半饱血雌蜱唾液腺cDNA文库的EST中筛选了1个含polyA尾的基因序列,经5LRACE方法得到该基因全长序列.经同源性比较,该基因预测的氨基酸序列与长角血蜱肌钙蛋白I(GI14041807)的同源性为84.47%,肌动蛋白结合位点位于147-167氨基酸处,且与长角血蜱肌钙蛋白I肌动蛋白结合位点完全相同,表明该基因是镰形扇头蜱肌钙蛋白I基因.以镰形扇头蜱基因组DNA为模板扩增到编码肌钙蛋白I的基因组序列.序列分析表明该序列不含内含子.RT-PCR分析表明该基因在镰形扇头蜱的卵及其幼蜱、若蜱、成蜱的壳、唾液腺和肠均有表达.     

镰形扇头蜱肌钙蛋白Ⅰ基因的克隆及其在大肠杆菌中的表达

目的克隆镰形扇头蜱肌钙蛋白Ⅰ(TnI)的cDNA并进行表达及鉴定。方法用RT-PCR方法从镰形扇头蜱总RNA中克隆编码肌钙蛋白Ⅰ的cDNA片段,将该cDNA连接到pET-32a( )表达载体,并转入宿主菌BL21(DE3),经IPTG诱导表达,并对表达产物进行Western blot分析。结果获得镰形扇头蜱的eDNA,并在大肠杆菌中以包含体的形式高效表达,Westem blot分析表明,抗TnI重组蛋白兔血清可以识别蜱体内的TnI,并在22 kDa和36 kDa之间有两条带。结论蜱体内可能同时存在两个大小不等的TnI,为进一步的研究打下基础。     

微小牛蜱唾液菌群的PCR-DGGE分析

为了解微小牛蜱唾液内的菌群结构信息,采用PCR-DGGE技术对微小牛蜱半饱血、饱血雌成虫唾液内菌群结构进行了分析和比较,结果表明:微小牛蜱半饱血、饱血雌成虫唾液内菌群结构有较大差异,饱血中细菌比半饱血多两种,分别为莫拉菌和不动杆菌;唾液内菌群与全蜱菌群差异极大。在检出的细菌中多数具有一定的致病性,故微小牛蜱叮咬可能会引起人或动物发生多种细菌性疾病。     

褐黄血蜱cystatin基因全长的克隆及原核表达研究

为了探讨褐黄血蜱体内半胱氨酸蛋白抑制剂(cystatin)蛋白及其编码基因的情况,试验基于褐黄血蜱唾液腺转录组数据库中搜索的注释为cystatin的contig_43533序列设计引物,RACE扩增其5′端和3′端,拼接获得cDNA全长,利用生物信息学软件进行分析;以饱血褐黄血蜱雌成虫全蜱总cDNA为模板,扩增cystatin-ORF,构建重组表达载体,原核表达、纯化制备cystatin重组蛋白。结果表明:褐黄血蜱cystatin cDNA全长635 bp,包含396 bp的开放阅读框架,编码一个含131个氨基酸残基的蛋白,原核表达可获得褐黄血蜱cystatin重组蛋白。说明褐黄血蜱cystatin蛋白能在大肠杆菌内以可溶性形式稳定表达。     

不同饲喂方式和温度对长角血蜱甘肃株生物学特性的影响

在不同饲喂方式和温度条件下,观察我国北方长角血蜱(Haemaphysalis longicornis)甘肃株的生活史及对其生物学特性的影响。将试验蜱分组后置于不同饲喂方式和温度下,待其发育到下一阶段,叮咬实验动物,连续调查其发育阶段的生物学特性。研究发现,完成一个世代需101 d～148 d。成年饥饿雌蜱吸血期具有两个显著特征:缓慢吸血和快速吸血;经兔体叮咬的成年雌蜱其饱血体重小于羊体。不同饲喂条件下长角血蜱饱血体重、产卵量差异极显著(P0.01);在一定温度范围内,幼蜱、若蜱的蜕变时间随温度的升高而缩短;当饱血雌蜱处于发育阶段时,产卵前期、产卵期及卵的孵化期均随着温度的升高而缩短;16℃～30℃条件下长角血蜱产卵量没有变化,利于蜱的存活;4℃以下和45℃以上饱血蜱及其后代不能存活;16℃以下蜱的发育周期延长。结果表明,长角血蜱为三宿主蜱;雄蜱可以帮助雌蜱吸血;在羊体上的吸血能力优于在兔体上的吸血能力;温度对长角血蜱的发育繁殖有影响。     

Pulmonary atherosclerosis and pulmonary arterial pressure in cholesterol-fed New Zealand white rabbits.

The lung produces many vasoactive substances originating from its vascular endothelium and plays an important part in various pathose. The present study was carried out to clarify pulmonary atherosclerosis and pulmonary arterial pressure, and to elucidate a part of the pulmonary pathosis in cholesterol-fed rabbits. Atherosclerosis was induced by feeding the animals a cholesterol-rich diet. When the rabbits were fed the cholesterol-enriched diets for 15 weeks, the grade of the atherosclerosis was severer than in 8W-feeding rabbits. The lesions of 8W-feeding rabbits were mainly composed of foam cells and fibrous components, whereas in 15W-feeding rabbits, the aggregation of foam cells beneath the endothelium of the vessel was infiltrating the media and severe stenose of the lumen was observed. In the entire pulmonary arterial system, the severe obstructive vascular lesions were localized and not diffused. The pulmonary arterial pressures of the rabbits increased slightly with time and the mean pressures were 11.3+/-0.9 (control group), 11.8+/-1.0 (8W group) and 13.7+/-1.5 mmHg (15W group) respectively. A significant difference existed in the mean pressure between the control group and 15W-feeding group, but there were no significant differences in the systolic and diastolic pressures among the three groups. In conclusion, we could induce pulmonary atherosclerosis in rabbits by feeding them a hyper-cholesterol diet but not overt pulmonary hypertension.     

Influence of feeding intensity on the growth, body composition and sexual maturity of male New Zealand White rabbits

An experiment was carried out with young male New Zealand White (NZW) rabbits to establish live body weight changes, body measurements, body composition and sexual maturity as a function of feeding intensity. Animals in Group 1 ('AL', n = 10) were fed ad libitum, while those in Group 2 ('RS', n = 10) received restricted feeding corresponding to 70% of the ad libitum level. The starting liveweights were practically the same (0.907 +/- 0.146 and 0.911 +/- 0.147 kg in Group AL and Group RS, respectively). The feeding trial lasted from 6 to 22 weeks of age. The average body weight was significantly higher in Group AL from 7 to 22 weeks of age. At 22 weeks of age the body weight of RS rabbits was 85.64% of the weight of AL animals (3.22 +/- 0.52 kg and 3.76 +/- 0.33 kg, respectively). Average body weights of RS males at 8, 9, 11, 19 and 21 weeks of age were similar to those of ad libitum fed (AL) animals at 7, 8, 10, 15 and 16 weeks of age, respectively. The growth of bucks fed restricted tended to be allometric. The most significant difference was found at 16 and 18 weeks of age, while the lowest difference occurred at 6, 12, 15 and 19 weeks of age. It can be stated that low-intensity feeding up to slaughtering weight causes backwardness in rear cannon length and this backwardness remains also after the 15th week, which is well over the optimal slaughtering age. Based on the present data, the 70% restricted feeding cannot be recommended either for the future breeding bucks or for broiler males reared for slaughter. To determine the major chemical components of the body, rabbits were euthanised. Original dry matter and crude fat content of the body significantly (P < 0.05) decreased under restricted feeding (41.42%; 32.48% and 16.73%; 7.35%) while the percentage of protein within the dry matter increased (49.6%; 65.0%) and fat decreased (40.17%; 22.1%) significantly. Libido unambiguously decreases as a consequence of feed deprivation. The most conspicuous difference was found in the level of blood testosterone. Although a few RS bucks produced semen but only much later than the rabbits fed ad libitum. On the other hand, there was no difference in the motility of spermatozoa and ejaculate volume in comparison with AL animals. There was no relationship between the body fat content and the reproductive status of bucks in the present trial.     

Effect of feeding olive by-products on certain blood parameters and serum enzyme activities of fattening rabbits

The effect of feeding different levels (10 and 20%) of dehydrated olive cake pulp on certain blood parameters and on the catalytic activity of serum enzymes was studied in fattening rabbits. A total of 60 rabbits were divided into a control group (C) and two experimental groups (E1, E2), each comprising 10 males and 10 females. Rabbits of Group C received no dehydrated olive cake pulp in their diet, while rabbits of Group E1 and Group E2 were fed 10% and 20% dehydrated olive cake pulp, respectively. After Day 56 of the experiment, blood samples were obtained by cardiac puncture and certain blood parameters as well as the catalytic activity of serum enzymes were determined. Following the experimental feeding period no significant differences were found between Group C and Groups E1 and E2 in red blood cell (RBC) and white blood cell (WBC) count, haemoglobin concentration and haematocrit value. Similarly, after 56 days of experimental fattening no significant differences were found between the control group and the experimental groups in the catalytic activity of the serum enzymes alkaline phosphatase (AP), aspartate amino-transferase (AST), gamma-glutamyltransferase (GGT), and lactate dehydrogenase (LDH). However, Group E2 rabbits manifested a significantly higher alanine amino-transferase (ALT) activity in the serum than did rabbits of Group E1. The results indicate that the inclusion of dehydrated olive cake pulp in the diet at the rate of 10% or 20% caused no changes in the investigated blood parameters and serum enzyme activities of fattening rabbits.     

意大利黑麦草应用于肉兔养殖的研究

试验的目的是探讨以意大利黑麦草和野生杂草作为青饲料代替部分精饲料饲喂肉兔对其增重、屠宰性状及兔肉品质的影响。经过40 d的分组饲喂试验发现,利用野生杂草或意大利黑麦草代替10%～50%的精饲料饲喂肉兔对其增重效果无显著影响（P〉0.05）;青饲料的引入在一定程度上增加了肉兔的屠宰率,还明显降低了兔肉中的脂肪含量,改善了兔肉的品质。     

引入西藏高原的家兔血液生化参数的比较研究

对引入西藏高原多年的加利福尼亚兔和中国白兔的12项血清生化指标进行测定,并与SPF新西兰兔进行比较。结果显示：①3个品种公兔之间,加尼福利亚兔与中国白兔比较,AST、TP、ALB、GLO、A／G、GLU、CRE、LDH差异极显著（P〈0．01）,TG、BUN差异显著（P〈0．05）,其中TP、ALB、A／G、CRE、CHOL、LDH表现极强的公兔特征,而ALT、GLU母兔明显高于公兔;加利福尼亚兔与SPF新西兰兔比较。ALT、A／G、CRE、TG、BUN差异显著（P〈0．05）,AST、CHOL、LDH差异极显著（P〈0．01）;中国白兔与SPF新西兰兔比较,ALT、AST、TP、ALB、GLO、A／G、CRE、CHOL、TG差异极显著（P〈0．01）;②3个品种母兔之间,加利福尼亚兔与中国白兔比较,ALT、GLO、CRE、TG差异显著（P〈0．05）,AST、TP、BUN、LDH差异极显著（P〈0．01）,加利福尼亚兔与SPF新西兰兔比较,ALT、TP、ALB、GLO、A／G、BUN、CRE、LDH差异极显著（P〈0．01）,AST、TG差异显著（P〈0．05）;中国白兔与SPF新西兰兔比较,ALT、TP、ALB、GLO、A／G、CRE、CHOL、TG、LDH差异极显著（P〈0．01）,BUN、AST差异显著（P〈0．05）。     

益母生化散对八眉猪繁殖性能的影响效果

选用健康状况良好的经产八眉猪二元母猪(2～4胎次)120头,随机分成2组。试验组的处理方法为:母猪产后2～3h后饲喂益母生化散,随后相隔24～48h在进行第二次饲喂和第三次饲喂;对照组只注射一针抗生素(青霉素钾)药物之后,两组在相同的饲养管理条件下,进行跟踪观察两组母猪在下一繁殖周期中出现产科疾病猪头数,35d断奶至配种平均间隔天数、情期受胎率、产仔率及淘汰率等繁殖生产成绩。试验结果表明,试验组35d断奶至配种平均间隔天数、情期受胎率和产仔率比对照组有明显改善,差异显著(P<0.05).     

不同水平蚕沙对新西兰兔肉品质的影响

试验旨在研究不同水平蚕沙对新西兰兔肉品质的影响。选取120只健康、体重相近(1.06 kg±0.09 kg,公、母各半)的35日龄断奶新西兰兔,随机分为5组,每组6个重复,每个重复4只。对照组饲喂基础日粮,试验组分别饲喂添加6%、12%、18%和24%蚕沙的日粮。试验结束后,每组选取接近平均体重的4只兔(公、母各半)屠宰,测定肌肉品质指标。结果显示,不同水平蚕沙对新西兰兔肉品质有一定影响,其中各组肉色、pH、失水率和蒸煮率差异不显著(P＞0.05),而剪切值试验组显著低于对照组(P＜0.05),且随蚕沙添加量的增加呈减小的趋势;在化学性状中各组水分、粗蛋白质、粗脂肪、灰分含量差异不显著(P＞0.05)。     

Growth,behavioural, serum biochemical and morphological changes in female rabbits fed high-fat diet

This study aimed to determine whether high-fat diet (HFD) could cause growth, behavioural, biochemical and morphological changes in young female rabbits. Thirty-six female rabbits were randomly divided into two groups fed with either a high-fat diet (HFD) or a standard normal diet (SND) for 5 weeks. Growth and behavioural changes were recorded during the 5-week feeding period. Tissue samples, including blood and adipose tissue, were obtained after slaughter. HFD rabbits weighed more by the end of the feeding period, had a higher percent body weight and adipose tissue weight change and had longer body and bust lengths than SND rabbits. HFD rabbits significantly reduced their feed intake and feeding frequency during the fourth and fifth weeks. HFD rabbits also showed lower frequency of drinking and resting and increased stereotypical behaviour. Besides, HFD rabbits showed significant physiological abnormalities. HFD rabbits had higher serum cholesterol (TC) and triglycerides (TG) levels than SND rabbits at the end of the feeding period, and higher free fatty acid (FFA) levels than rabbits in the SND group after the third week of feeding. Serum thyroxine (T4) increased significantly in week 2 and week 5 and triiodothyronine (T3) increased significantly in week four. However, there was no significant change in serum glucose (GLU) and insulin (INS) levels. Additionally, HFD reduced the area and diameter of perirenal and subcutaneous fat cells and increased their density. Our findings suggest that HFD rabbits had higher weight gains, accumulation of fat, and more behavioural changes than SND rabbits. Although high levels of fat in the diet had a low impact on hyperglycaemia, it could lead to hyperlipidemia and hyperthyroidism. Our results also suggest that sustained HFD may cause the proliferation of adipocytes in young female rabbits.     

扶正解毒散对兔的生产性能及血液生理指标的影响

为了获得增强家兔非特异性保护屏障的措施和保健方案,选择1月龄500g左右的健康新西兰兔6只,分成试验组和空白对照组饲喂一个月。于试验前、试验结束时分别采集各试验兔的血液和称取体重,检测血液主要生理指标,并作统计分析。结果表明:添加并饲喂扶正解毒散的试验组兔的血液主要生理指标较试验前大幅增加,统计分析P<0.01,差异极显著;血液主要生理指标试验后组间差异性t-检验:双样本方差分析得出,白细胞量P值<0.01,差异极显著;红细胞、血红蛋白、平均血红蛋白浓度、平均血红蛋白量的P值:0.01相似文献   

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