城市废水暴露对食蚊鱼肝脏EROD酶活性的影响

采用动力学酶标荧光法,检测了东莞市数所污水处理厂、制药厂和电子厂废水对食蚊鱼(Gambusia affinis)肝组织中7-ethoxyresorufin o-deethylase(EROD)酶活性的影响,评价了运用EROD酶活性监测水环境污染物的生物效应的可行性。结果显示,食蚊鱼分别暴露于经稀释为20%,40%,60%,80%不同梯度的废水液72 h后,肝脏EROD酶的活性分别与受试城市污水处理厂、制药厂和电子厂的废水之间存在剂量效应关系,EROD酶活性随污水浓度的增加而提高。电子厂废水的最大诱导倍数与对照组的比值可达到5.26,这表明其水体中存在的有机污染物较多,污水处理厂次之,制药厂的出水中污染物最少。研究表明,食蚊鱼肝组织EROD酶活性可以作为监测城市废水污染的理想生物标记物,后续的研究工作应使之标准化。     

The fate of chlortetracycline residues in a simulated chicken–fish integrated farming systems

Two experiments were conducted at the Asian Institute of Technology, Pathumthani, Thailand to investigate the fate of chlortetracycline (CTC) residue in chicken manure and its effect on integrated chicken–fish farming system. During the first experiment, broiler chickens were raised and CTC residues in their manure were analysed. Chicken fed diets containing 0, 50, 200 and 800 CTC mg kg^?1 had CTC residue levels of 0, 0.9, 3.8 and 6.5 CTC ng g^?1. Once the diet containing CTC was withdrawn, CTC in the manure dropped to negligible amounts (0, 0, 0.2 and 0.5 CTC ng g^?1) within 1 day. Integrated chicken–fish farming systems were simulated during the second experiment to determine the fate of antibiotic residues in chicken manure in aquaculture environment. Chickens were fed a CTC‐free diet and a feed containing CTC at 200 mg kg^?1. Ten 4 m³ square concrete tanks (2 × 2 × 1 m) were used for the experiment. Five tanks were fertilized with CTC‐contaminated manure and the remaining five tanks were fertilized with CTC‐free manure at a rate of 100 kg dry matter ha^?1 day^?1. Sex‐reversed Nile tilapia (Oreochromis niloticus) was stocked at 12 fish tank^?1 on the 14th day after chicken manure application. The immuno‐radio microbial receptor assay (Charm II test) revealed that edible fish muscle, fish intestinal tract and sediment were contaminated by CTC at rates of 7.21, 22.104 and 1.788 ng g^?1, respectively, after 45 days. Chlortetracycline was detected on day 20 in the water column and gradually increased from 0.26 to 12.13 ng g^?1. Chlortetracycline residues were not detected in fish or the aquatic environment of the CTC‐free treatment. The results demonstrate the potential for antibiotic residue accumulation in fish and aquatic environment when CTC‐contaminated chicken manure is used for pond fertilization.     

Analysis of exotoxins produced by atypical isolates of Aeromonas salmonicida,by enzymatic and serological methods

In this study, exotoxins produced by 62 Aeromonas salmonicida strains and the bacterium Haemophilus piscium were analysed. Enzymatic assays, zymograms and serological detection were used to monitor secretion by bacterial strains of the previously described exotoxins P1, GCAT and AsaP1 and also the extracellular P2 metallo-gelatinase and a serine caseinase, which is different from the P1 protease and has not yet been characterized. Based on the results, the strains were divided into five groups. One comprised the type strains for A. salmonicida ssp. masoucida, H. piscium and 36% of the atypical isolates, and another, a type strain for A. salmonicida ssp. smithia together with 14% of the atypical isolates. A second type strain of A. salmonicida ssp. smithia was grouped with 8% of the atypical isolates. The largest group contained the type strains for A. salmonicida ssp. achromogenes and 38% of the atypical isolates. The type strains for A. salmonicida ssp. salmonicida were in the last group with all the four typical strains and 4% of the atypical isolates. The combination of zymogram and serological detection used is recommended as the most reliable method for characterizing A. salmonicida strains according to their exotoxin secretion.     

口蹄疫灭活疫苗蛋白质含量测定操作程序的确立

为优化用于口蹄疫灭活疫苗蛋白质含量测定的改良Lowry法,进而确立口蹄疫灭活疫苗蛋白质含量测定的操作程序,探索了有机溶剂破乳剂、酚红以及丙酮沉淀对测定结果影响。结果表明：样品中含酚红和有机溶剂均导致测定值较标准值高;有机溶剂破乳后,水相样经过丙酮沉淀测定值较标准值低;丙酮直接沉淀疫苗后测定蛋白质值与标准值符合度最高,丙酮沉淀回收率随蛋白浓度升高而升高,回收率在90％～100％之间。试验首次确立了改良Lowry法检测口蹄疫灭活疫苗中蛋白质含量的操作程序为丙酮直接沉淀疫苗后测定蛋白质浓度。并成功应用于口蹄疫灭活疫苗蛋白质含量的测定。     

重庆市禽白血病及禽网状内皮组织增生症的血清流行病学调查

禽白血病和禽网状内皮组织增生症均为禽的肿瘤性免疫抑制疾病,是危害养鸡业的两种非常重要的病毒性传染病。为调查重庆市肉鸡中禽白血病及禽网状内皮组织增生症的流行情况,在北碚区、涪陵区、开县、垫江县、潼南县五个区（县）的8个活禽交易市场采集260份血清样本,采用酶联免疫吸附试验检测了所有血清中禽白血病病毒（ALV）和禽网状内皮组织增生病毒（REV）的抗体。检测结果显示：禽白血病病毒和禽网状内皮组织增生病毒抗体阳性率分别为16.5%（43/260）和5%（13/260）,双抗体阳性率为4.61%（12/260）。与全国其他地区相比,重庆市肉鸡中这两种病原的感染率相对要低,但仍应重视这两种疾病的防控,以控制病原的进一步传播。     

应用酵母双杂交方法筛选与猪瘟病毒Npro蛋白相互作用的宿主蛋白

为研究猪瘟病毒(CSFV)与宿主细胞间的相互作用,本研究采用酵母双杂交方法以猪瘟病毒Npro蛋白为诱饵,从猪外周血单个核细胞(PBMC) cDNA表达文库中筛选与之相互作用的蛋白,经共转化试验表明,共筛选到12个与Npro蛋白相互作用的宿主蛋白,应用Cytoscape 2.8.2软件绘制了蛋白质间相互作用关系图,根据GO分析结果,这些蛋白分别参与细胞代谢、细胞生长、免疫等过程,经双分子荧光互补(BiFC)试验表明筛得的部分宿主蛋白与CSFV Npro具有相互作用.本研究中筛选得到的蛋白质对CSFV复制的影响有待进一步深入研究.     

Phage and MLVA Typing of Salmonella Enteritidis Isolated from Layers and Humans in Belgium from 2000–2010, A Period in which Vaccination of Laying Hens was Introduced

The aim of the study was to characterize isolates of Salmonella enterica serovar Enteritidis (S. Enteritidis) obtained from humans and layer farms in Belgium collected during 2000–2010. Three periods were compared, namely (i) before implementation of vaccination (2000–2004), (ii) during voluntary vaccination (2005–2006) and (iii) during implementation of the national control program (NCP) for Salmonella including mandatory vaccination against S. Enteritidis (2007–2010). The characteristics compared across time periods were distributions of phage type and multiple‐locus variable number tandem‐repeat assay (MLVA). While PT4 and PT21 were predominantly isolated in Belgium in layers and humans before 2007, a significant reduction of those PTs was observed in both populations in the period 2007–2010. The relative proportion of PT4b, PT21c and PT6c was found to have increased considerably in the layer population since 2007. In the human population, PT8, PT1 and the group of ‘other’ PTs were more frequently isolated compared to the previous periods. When comparing the proportion of the predominant MLVA types Q2 and U2, no significant difference was found between the layer and human population in the three periods and between periods within each category (layer and human). A significant difference in isolate distribution among MLVA clusters I and II was found between human and layer isolates recovered during Period 3 and in the human population between Period 1 and 3. Results suggest that the association between S. Enteritidis in layers and the occurrence of the pathogen in humans changed since implementation of the NCP in 2007.     

用酶联免疫吸附受体法检测鱼类生长激素的生物活性

根据激素一受体反应的原理，结合酶联免疫吸附测定法（ＥＬＩＳＡ）和放射受体测定法（ＲＲＡ）的优点，应用我们纯化的Ａ昌鱼生长激素（ｇｃＧＨ）和大鳞大马哈鱼生长激素（ｓＧＨ）及其特异抗体，采用鱼类肝细胞膜受体制剂，首次建立了测定鱼类ＧＨ生物活性的酶联免疫吸附受体测定法（ＥＬＩＳＡ－ＲＡ）。此法检测草鱼ＧＨ的灵敏度达０．０６３￣０．１２５ｕｇ／ｍｌ，检测大马哈鱼ＧＨ与草鱼肝膜受体结合的灵敏度达０．２５ｕｇ     

A rapid immunoperoxidase-based virus neutralization assay for salmonid alphavirus used for a serological survey in Northern Ireland

A modified virus neutralization (VN) assay was developed to replace an existing assay read on the presence or absence of virus-induced cytopathic effect (CPE). The modified assay used a monoclonal antibody to salmon pancreas disease virus as the first layer of an immunoperoxidase (IPX)-based immunostaining technique to detect viral growth. The IPX-based VN assay required only 3 days to perform, and the adoption of a 96-well microtitre format facilitated a high throughput of samples requiring small volumes of serum, cells and virus. When 352 sera from farmed salmon and 302 sera from farmed trout were tested by both the modified and the original CPE-based assays, overall correlations of 97.72 and 96.03% were, respectively, obtained (96.94% combined). When the modified assay was used to test 188 sera collected from wild salmonids in freshwater river systems in Northern Ireland, no positive results were recorded.     

Epidermal proteases of the Japanese eel

A fluorescent-sensitive assay was used to demonstrate the protease activity in the dorsal skin of Japanese eel (Anguilla japonica). Two distinct extracts were separately prepared from skin mucus and epidermal cell layers, with no mutual contamination. The epidermal extract was sensitive to various substrates, whereas there was no, or only marginal, susceptibility to the same substrates for the mucous extract. Optimum hydrolysis pHs of the epidermal extract was variable and below pH 7.0, and the optimum hydrolysis temperatures were between 40 and 50 °C. In addition, Tos-Phe-Ch₂Cl, chymostatin, CdCl₂, CuCl₂, HgCl₂ and ZnCl₂ inhibited protease activities to different extents. Several other reagents specifically affected the protease activities, and their induced effects were useful for the identification of epidermal proteases. The findings indicate that a proteolytic factor, exhibiting various enzymological specificities, is retained within epidermal cell layers of Japanese eel. This factor is composed of 4 distinct proteases, such as cathepsins L and B-like proteases, a serine protease and an aminopeptidase.