4种狂犬病病毒ELISA抗体检测试剂盒的评价

为评价国产狂犬病病毒ELISA抗体检测试剂盒的性能,选择4种国产品牌ELISA抗体检测试剂盒,分别检测463份已用荧光抗体病毒中和试验（FAVN）测知狂犬病病毒抗体效价的犬血清,用Kappa检验和配对卡方检验评价ELISA试剂盒与FAVN检测结果的一致性和差异性,计算ELISA试剂盒的诊断敏感性、诊断特异性、符合率和重复性等,并比较4种试剂盒的使用范围、样品稀释倍数等参数。结果显示：经Kappa检验,4种试剂盒与FAVN一致性均为弱;经配对卡方检验,A、B试剂盒与FAVN检测结果的差异不显著（P> 0.05）,而C、D试剂盒差异显著（P <0.05）。诊断敏感性,D试剂盒最高,B试剂盒最低;4种试剂盒诊断特异性均较低,为29.5%～57.7%;各试剂盒符合率相当,为71.3%～72.8%。综合敏感性、特异性、重复性等考量因素,得出C试剂盒更能满足免疫后抗体检测需求,A试剂盒也较好,两者可作为候选试剂盒。结果表明,4种国产狂犬病病毒ELISA试剂盒的诊断敏感性、诊断特异性、批间稳定性等性能需进一步提高,以满足基层免疫抗体监测需求。     

苏云金芽孢杆菌Cry8Ca2蛋白的纯化与活性的研究

cry8 Ca2基因是本实验室自行分离克隆的一种新型苏云金芽孢杆菌(Bacillus thuringiensis,Bt)杀虫晶体蛋白基因,其表达130 kDa蛋白对铜绿异丽金龟(Anomala corpulenta)和黄褐异丽金龟(A.exoleta)幼虫具有较高的活性。本研究对Cry8Ca2蛋白原毒素及胰凝乳蛋白酶消化片断的纯化条件及活性进行研究,最适的消化条件为质量比1∶20,37℃,消化1 h。通过阴离子交换层析得到纯的活性毒素蛋白。用原毒素与纯化的毒素对铜绿丽金龟幼虫进行生测,Cry8Ca2原毒素蛋白的LC₅₀为0.47μg/g土,纯化的毒素的LC50为0.08μg/g土,毒素的杀虫活性可以达到原毒素的6倍。     

茶树病程相关蛋白基因CsPR5的克隆及表达分析

为探究茶树中病程相关蛋白5(pathogenesis-related protein 5,PR5)在茶树中的功能,采用cDNA末端快速扩增(rapid amplification of cDNA end,RACE)技术克隆了PR5基因的全长;通过荧光定量PCR方法检测了CsPR5在龙井43茶树不同组织部位、植物激素(茉莉酸、乙烯利和水杨酸)处理、茶炭疽病菌Colletotrichum camelliae侵染和小贯小绿叶蝉Empoasca onukii为害后的表达特征。结果表明,CsPR5基因开放阅读框为843 bp,编码281个氨基酸。CsPR5基因在茶树根、茎、嫩叶、花和种子5个组织中的相对表达量分别为0.187、3.093、0.928、0.045和0.012,且五者之间差异显著,主要在茎和叶中表达,其分别为根中相对表达量的16.54倍与4.96倍。水杨酸处理6 h及乙烯利处理6、12 h后,茶树CsPR5基因的相对表达量分别为1.622、2.344和1.845,分别比对照显著上调2.20倍、3.18倍和2.35倍;但是外施茉莉酸对茶树CsPR5基因的相对表达量无影响。茶炭疽病菌侵染显著诱导了茶树CsPR5基因的相对表达量,处理后的相对表达量为1.977,是对照的1.90倍;小贯小绿叶蝉取食显著抑制了其为害部位CsPR5的相对表达量,为7.273,仅为对照的38.80%,但该诱导不具系统性。     

小麦水分和装满度对磷化氢吸附率的影响

在小麦含水量分别为9.3%、12.6%和14.4%,装满度分别为100%、70%、50%、30%和0%,气体浓度为100mL/m3和400mL/m3的条件下,测定小麦对磷化氢的吸附率,结果表明小麦对磷化氢的吸附率随着粮食水分增加,磷化氢浓度和装满度增大有不同程度增加.在药量较大的施药初期和装满度大时,检测到的磷化氢浓度高于设定浓度,磷化氢浓度、小麦水分和装满度较低时,小麦对磷化氢的吸附较容易达到平衡.     

Immunomodulatory effect of γ‐irradiated Astragalus polysaccharides on immunosuppressed broilers

In this study, we irradiated Astragalus polysaccharides (APS) using 25 kGy ⁶⁰Co γ ray to obtain γ‐irradiated Astragalus polysaccharides (IAPS) and then investigated the effects of IAPS on growth performance and immune function of cyclophosphamide (CPM)‐treated broilers. The physicochemical properties of APS and IAPS (molecular weight, water solubility, viscosity, morphological and structural properties) were evaluated. Then, 384 one‐day‐old Arbor Acres broiler chicks with similar initial weight were randomly assigned into 6 groups: the non‐treated group (control), and CPM‐treated groups were fed either a basal diet or the diets containing 900 mg/kg APS, or 900, 600, 300 mg/kg IAPS, respectively. On days 16, 18, and 20, all broilers except for the control group were intramuscularly injected with 0.5 ml CPM (40 mg/kg·BW). Broilers in the control group were intramuscularly injected with 0.5 ml sterilized saline (0.75%, wt/vol). This trial lasted for 21 days. The physicochemical treatment showed that γ irradiation could decrease the molecular weight and viscosity, and increase the water solubility of APS (p < 0.05), whereas the structural properties of APS was not affected. In the animal trial, 900 mg/kg APS or 900, 600 mg/kg IAPS relieved the decreased growth performance, thymus index, T lymphocytes proliferation, serum IgG concentration, NOS activity and the increased blood heterophil:lymphocyte ratio in CPM‐treated broilers (p < 0.05). CPM‐induced decreases in B lymphocytes proliferation and serum IgM concentration were only increased by IAPS at 900 mg/kg (p < 0.05). Overall, both APS and IAPS alleviated CPM‐induced immunosuppression. Especially, IAPS possessed better immunomodulatory effect than APS, indicating that γ irradiation could be used as an effective method to enhance the immunomodulatory activity of APS.     

梨不同砧穗组合光合特性

 用CIRAS-1 便携式光合作用测定系统研究了豆梨(Pyrus betulaefolia Bunge) 、杜梨(P.calleryana Dcen.) 幼树以及豆梨、杜梨与砂梨(P. pyrifolia Nakai) 品种丰水、新高4 种砧穗组合幼树叶片的光合作用。结果表明, 砧木显著影响栽培品种的光合作用, 杜梨比豆梨以及以杜梨为砧木的丰水、新高幼树比以豆梨为砧木的两品种幼树叶片净光合速率(Pn) 、表观量子产量(Φi) 、羧化效率(CE) 、水分利用效率(WUE) 高。新高的Pn、Φi 、CE 和WUE 值较丰水高, 且在两种砧木上均表现出同样的结果。     

表达马立克氏病病毒CVI988／Rispens株糖蛋白B基因的重组鸡痘病毒疫苗的免疫保护研究

用鸡痘病毒载体表达马立克氏病病毒（MDV）糖蛋白B（gB）基因构建成重组鸡痘病毒（rFPV）。以MDV　GA或Md5和RBIB混合攻毒，在不同品种的鸡中评价rFPV－gB／R单价苗和与火鸡疱疹病毒（HVT）组成的二价苗的免疫保护效力。试验结果表明，MDV疫苗和FPV母源抗体阴性的SPF鸡和母源抗体阳性的商品鸡中，rFDV－gB／R均能提供免疫保护作用，且其中一种商品蛋鸡中rFPV－gB／R与HVT液氮苗或HVT冻干苗结合使用，均有显著的免疫协同保护作用。免疫学研究指出，rFPV－gB／R与常规疫苗一样，可显著地降低疫苗免疫攻毒鸡的病毒血症和羽囊排毒。     

Crop milk protein is synthesised following activation of the IRS1/Akt/TOR signalling pathway in the domestic pigeon (Columba livia)

1. The experiment was conducted to study whether insulin receptor substance 1 (IRS1) / Protein kinase B (Akt)/target of the rapamycin (TOR) signalling pathway activation stimulates crop milk protein synthesis in the domestic pigeon (Columba livia).

2. Crop milk was collected from ten 1-d-old squabs and analysed for nutrient content. During the non-breeding period and the first day of lactation, blood samples were collected from 5 pairs of breeding pigeons and the levels of prolactin and insulin were determined.

3. Crop samples were collected from 5 pairs of breeders at d 14 and 16 of the incubation period and d 1, 3 and 7 of the lactation period. Crop samples were evaluated for changes in crop weight and thickness and changes in the expression patterns of IRS1/Akt/TOR signalling pathway–related proteins.

4. The results demonstrated that prolactin induces a gradual increase in the relative weight and thickness of the crop, with crops reaching a maximum size at the third day of lactation.

5. Pigeon crop milk contains 64.1% crude protein and 29.7% crude fat based on dry weight.

6. Serum prolactin and insulin levels in the lactation period were significantly higher than those in the non-breeding period. Compared with non-breeding pigeons, the expression of the phosphorylated IRS1 phosphorylated Akt, phosphorylated TOR, phosphorylated ribosomal protein S6 kinase, phosphorylated S6, phosphorylated eukaryotic initiation factor 4E binding protein 1 and eukaryotic initiation factor 4E were significantly up-regulated in the crop of pigeons in the lactation period.

7. In conclusion, prolactin might induce changes in crop tissue and form the physiological structure for crop milk synthesis. Furthermore, the synthesis of crop milk protein is regulated by activation of the IRS1/Akt/TOR signalling pathway.

     

小麦矮腥黑粉菌g9890基因编码效应蛋白的生物信息学分析及亚细胞定位

为明确小麦矮腥黑粉菌Tilletia controversa g9890基因编码效应蛋白的生物学功能,根据小麦矮腥黑粉病菌转录组测序结果,筛选出效应蛋白g9890,通过PCR技术获得g9890基因cDNA的全长序列,并对其进行生物信息学以及亚细胞定位分析。多种生物信息学数据库分析表明,g9890基因全长为1 038 bp（包括终止密码子）,共编码345个氨基酸,相对分子质量为37 353.32,理论等电点为5.02。g9890效应蛋白不稳定系数为15.94,疏水性指数为-0.312,是一种亲水性且稳定的蛋白。将g9890基因与pbin-GFP载体重组,利用冻融法转化至根癌农杆菌Agrobacterium tumefaciens GV3101中,将其注入烟草进行瞬时表达分析,并通过共聚焦激光显微镜观测该基因的定位状况,结果显示,g9890定位在细胞膜和细胞核上。     

山东省沂南县动物强制免疫“先打后补”现状分析

2021年,沂南县推进动物疫病强制免疫“先打后补”工作,依托“鲁牧云”信息平台,通过强化组织领导,加强政策宣贯,强化人员培训,放开强免疫苗经营渠道,提供抗体检测无偿服务等措施,提升了养殖场户“先打后补”的积极性、自主性、灵活性,强化了养殖场户动物防疫主体责任,满足了养殖场户疫苗多样化需求。但由于 “先打后补”补助金额少、社会化服务不规范、基层动物防疫体系弱化、缺少配套工作经费等原因,部分养殖场户实施“先打后补”的积极性不高。建议通过规范社会化服务组织,强化培训水平,提高补助标准,强化三级（县乡村）基础设施建设,升级“鲁牧云”等措施,确保“先打后补”政策惠及更多养殖场户。