草鱼外周血白细胞组成及其受胁迫和感染的影响

为了得到可反映草鱼（Ctenopharyngodon idellus）健康状况的血细胞指标的基础数据,对4 种不同质量规 格的草鱼进行白细胞分类计数研究,并观察草鱼在硫酸铜、敌百虫处理以及多子小瓜虫感染后白细胞组成的变化。 结果表明,1 000 g组和500 g组规格草鱼的淋巴细胞比例显著高于250 g组,1 000 g组和500 g组规格草鱼的中性 粒细胞比例显著低于60 g组和250 g组。硫酸铜处理组的淋巴细胞分类计数显著高于对照组;高浓度敌百虫处理组 草鱼的淋巴和血栓细胞分类计数显著高于对照组;多子小瓜虫感染组草鱼中性粒细胞分类计数极显著下降,淋巴细 胞分类计数显著上升。说明个体大小、理化胁迫因子和病原感染都会影响草鱼白细胞组成比例。     

不同淋洗剂对污染土娄土中Cu、Pb的淋洗修复研究

【目的】研究不同条件下螯合剂乙二胺四乙酸二钠盐(EDTA)、柠檬酸(CIT)和阴离子表面活性剂十二烷基硫酸钠(SDS)、生物表面活性剂鼠李糖脂(RL2)对污染土娄土中铜、铅的淋洗效果,并对淋洗机制进行探讨。【方法】以铜、铅单一污染的土娄土为研究对象,分别加入2种螯合剂(EDTA、CIT)和2种表面活性剂(SDS、RL2),采用批次试验方法,分析淋洗剂浓度、pH、背景电解质(NaNO3和NaCl)、水(mL)土(g)比、淋洗次数对污染土娄土中Cu和Pb的淋洗百分率的影响,探索不同淋洗剂淋洗污染土娄土中Cu和Pb的最佳条件。【结果】在相同浓度下,EDTA和CIT对污染土娄土中Cu的淋洗百分率明显高于其他淋洗剂,EDTA对Pb的淋洗百分率明显高于其他淋洗剂。淋洗4次后,4种淋洗剂对污染土娄土中Cu的淋洗能力由大到小为CITEDTARL2SDS,对污染土娄土中Pb的淋洗能力为EDTARL2CITSDS。EDTA和CIT溶液淋洗土娄土中Cu和Pb的最适条件为:水土比20(mL)∶1(g)、淋洗剂浓度0.01mol/L、pH=7、无背景电解质;当水土比为20(mL)∶1(g)、RL2溶液浓度为0.02mol/L、pH=8、无背景电解质条件下,RL2对污染土娄土中Cu、Pb的淋洗效果最佳。【结论】EDTA、CIT和RL2可以有效去除污染土娄土中的Cu,EDTA可作为石灰性污染土娄土中Pb的最佳淋洗剂,而SDS不宜作为污染性土壤的淋洗剂。     

Antibody-coated gold nanoparticles immunoassay for direct detection of Aeromonas salmonicida in fish tissues

Aeromonas salmonicida is the causative agent of furunculosis, a disease that affects both salmonid and non‐salmonid fish. Detection of A. salmonicida can be labour intensive and time consuming because of the difficulties in distinguishing the bacterium from other species given the wide variety of existing biochemical profiles and the slow growth characteristics which allow other organisms to overgrow the A. salmonicida. Herein, we report the development of a specific immunoassay using gold‐conjugated polyclonal antibodies for the rapid detection of A. salmonicida in fish tissues. Monodispersible 13‐nm gold nanoparticles were coated with polyclonal antibodies specific to A. salmonicida. Reddish purple agglutination of gold particles indicated the presence of A. salmonicida in samples. Positive reactions were detected visually with the naked eye. No agglutination was observed when A. salmonicida antibody‐coated gold nanoparticles were tested with other common bacterial fish pathogens, thereby verifying the specificity of the assay. The assay could detect A. salmonicida in fish tissues down to 1 × 10⁴ CFU mL^?1, and results were obtained within 45 min. The antibody‐coated gold nanoparticles were stable for at least 2 months at 4°C. The immunoassay using antibody‐coated gold nanoparticles represents a promising tool for the rapid and specific detection of A. salmonicida in fish tissues.     

Effect of calcium and magnesium hardness on acute copper toxicity to Indian major carp, Labeo rohita (Hamilton) and catfish, Channa punctatus (Bloch)

Four experiments were conducted to evaluate the effects of calcium and magnesium hardness on the acute toxicity of copper sulphate to Indian major carp, rohu (Labeo rohita, Hamilton) fingerlings and juvenile catfish (Channa punctatus, Bloch) in medium alkalinity experiments. A preliminary bioassay determined the 96 h LC₅₀ of copper sulphate to be 0.56 mg L^?1 for L. rohita fingerlings and 11.78 mg L^?1 for juvenile C. punctatus placed in water with calcium hardness and total alkalinity set at 100 mg L^?1 CaCO₃. In the first experiment, rohu were exposed to 0.56 mg L^?1 copper sulphate in environments where calcium hardness was varied from 50 to 350 mg L^?1 CaCO₃ and total alkalinity was 100 mg L^?1 CaCO₃. As calcium hardness increased, copper‐induced rohu mortalities decreased significantly from 90% at 50 mg L^?1 CaCO₃ to 7% at 350 mg L^?1 CaCO₃. In the second experiment, rohu were exposed to 0.56 mg L^?1 copper sulphate in environments where magnesium hardness was varied from 50 to 350 mg L^?1 CaCO₃ with total alkalinity set at 100 mg L^?1 CaCO₃. Hundred percent mortality was observed in magnesium‐based hardness treatments. In the third experiment, catfish were exposed to 11.78 mg L^?1 copper sulphate in environments where calcium hardness was varied from 50 to 400 mg L^?1 and total alkalinity was 100 mg L^?1 CaCO₃. As calcium hardness increased, copper‐induced catfish mortalities decreased significantly from 90% at 50 mg L^?1 CaCO₃ to 4% at 400 mg L^?1 CaCO₃. In the fourth experiment, catfish were exposed to 11.78 mg L^?1 copper sulphate in environments where magnesium hardness was varied from 50 to 400 mg L^?1 CaCO₃, with total alkalinity set at 100 mg L^?1 CaCO₃. In this case, 100% mortality was also observed in magnesium‐based treatments. Mortality rates in magnesium hardness treatments were consistent with those in the second experiment. These data suggest a calcium‐specific mechanism with respect to acute copper toxicity both in rohu and catfish.     

Assessment of the effect of silver nanoparticles on the relevant soil protozoan genus Acanthamoeba

Silver nanoparticles (AgNP) are used in a broad range of consumer products and industrial applications. During the regular product life cycle and disposal, AgNP are continuously released into the environment. Hence, the aim of this study was to investigate the potential ecotoxicological effects of AgNP exposure on amoebae. The Acanthamoeba castellanii ATCC 30234 strain and environmental isolate Acanthamoeba strain C5/2, which are both affiliated with genotype T4, were chosen as representatives of ecologically important soil protozoan organisms. The amoebae were exposed to citrate‐stabilized AgNP (30 and 70 nm in size) for 24 h and 96 h at concentrations ranging from 600 µg L⁻¹ to 20 mg L⁻¹. A newly adopted cell culture based microscopic assay was applied to assess the adherence ability of the amoeba trophozoites. The general metabolic activity of Acanthamoeba was determined to be a second independent endpoint by means of intracellular reduction of the redox dye AlamarBlue®. The fate of AgNP within the amoebae and test solutions was visualized by light‐ and transmission electron microscopy (TEM). Both Acanthamoeba strains showed a significant dose‐dependent decrease of adherence ability (p  < 0.04) and metabolic activity (p < 0.01) after 96 h of AgNP exposure. The environmental Acanthamoeba strain C5/2 lost both its adherence ability and metabolic activity at lower AgNP concentrations than the type strain, indicating a higher sensitivity to ionic silver. This was confirmed by the application of AgNO₃, provoking a higher effect level in strain C5/2. AgNP was visualized intracellularly by transmission electron microscopy within the cytoplasm of Acanthamoeba . This is the first report to show the ecotoxicological effects of short‐term AgNP exposure on the soil protist Acanthamoeba , causing both changes in the adherence ability and metabolic activity of this amoeba. This combined approach may be a powerful tool in the future for predicting potential harmful ecotoxicological effects of AgNP exposure using soil protozoans.     

奶牛集约化生产体系中铜素产污系数的测定

通过对集约化奶牛场的育成牛和泌乳牛各季节的粪尿产生量进行测定,并且分析粪样和尿样,从而计算出育成牛和泌乳牛的铜产污系数,并比较两者各季节间的差异性。结果显示集约化奶牛场的育成牛铜产污系数4个季节平均为0.14g/(head.d),泌乳牛铜产污系数4个季节平均为0.31g/(head.d),泌乳牛铜产污系数远大于育成牛铜产污系数。     

Physiological Responses of Wheat (Triticum Aestivum L.) –Aegilops Sharonensis Introgression Lines to Excess Copper

Two wheat lines (TL3 and TL5) derived from selection for Cu tolerance among wheat – Aegilops sharonensis hybrids were compared when grown as hydroponics at zero (control), 10⁻⁶ and 10⁻⁵  m CuSO₄. The morphometric measurements showed that the line TL5 with a more inhibited root system under 10⁻⁶  m CuSO₄ had a less inhibited shoot growth under 10⁻⁵  m CuSO₄ as compared to TL3. The decreased maximum efficiency of photosystem II and the more pronounced trend towards increased catalase activity suggested that despite the better shoot growth, the leaves of TL5 were more functionally injured. The increased content of carotenoids at excess Cu and the higher peroxidase and catalase activities of TL3 in control plants might contribute to its better stress tolerance. While no significant changes in enzyme activities were found at 10⁻⁶  m Cu, at 10⁻⁵  m the activities tended to increase. Although the close values of free phenolics concentrations in control plants, at Cu excess their content was higher in TL5 compared to TL3. The free phenolics content in roots at 10⁻⁶  m Cu decreased, and although higher at 10⁻⁵  m Cu, it remained below the control in TL3, and above the control in TL5. From the obtained results TL3 emerged to be more tolerant to excess Cu than TL5.     

肉鸡对三碱基氯化铜、铜氨基酸螯合物、铜蛋白盐生物利用率研究

采用玉米—豆粕型饲粮研究肉公雏对三碱基氯化铜(TBCC)、铜氨基酸螯合物(CuAA)、铜蛋白盐(CuPro)或硫酸铜(CuSO4)的生物利用率。在基础饲粮(Cu14mg/kg,DM基础 ,实测值)上 ,TBCC、CuAA、CuPro、CuSO4 的铜添加水平依次为150mg/kg、300mg/kg、450mg/kg,试验3周。结果表明 ,铜源和铜水平对试鸡体重和饲料转化率的影响不显著。以CuSO4 为参比标准 ,铜添加水平或日粮铜摄入量为自变量 ,经对数转换后肝铜积累为因变量建立多元线性回归模型 ,按斜率法综合求测铜源生物利用率为 :CuSO4 100 % ;TBCC112.4 % ;CuAA127.7 %(P<0.01) ;CuPro99.3 %。结果表明TBCC或CuPro生物利用率与CuSO4同效 ,但CuAA生物利用率比CuSO4 高。该试验提供了评定肉鸡有机铜源生物利用率的实用方法。