缓沉性水产膨化饲料加工工艺参数优化

该研究旨在研究不同加工工艺参数对缓沉性水产膨化饲料品质指标的影响规律,优化缓沉性水产膨化饲料的加工工艺参数。采用中心组合试验设计,选取调质物料含水率、膨化机螺杆转速和模头温度3个因素,吨料开孔面积固定为450 mm2/(t·h),调质物料含水率在23%～27%,模头温度范围在100～140 ℃,膨化机螺杆转速范围在180～300 r/min,对加工工艺参数进行优化。试验结果表明：随着调质物料含水率的升高,膨化饲料的容重逐渐降低,膨化率逐渐升高,在螺杆转速相同的条件下,不同模头温度的沉降速度的差异逐渐增大。随着模头温度的升高,膨化饲料的容重逐渐降低,膨化率逐渐升高,沉降速度逐渐降低,10 min沉水率先升高后降低。随着螺杆转速的增大,能耗逐渐升高,在调质物料含水率为23%～25%时,初始沉水率逐渐增大;当调质物料含水率大于25%时,10 min沉水率呈现先减小后增大的趋势。调质物料含水率和螺杆转速的交互项与初始沉水率呈负相关。调质物料含水率和模头温度的交互项与10 min内下沉率呈负相关。优化工艺参数：调质物料含水率为27%,模头温度110.68 ℃,螺杆转速265 r/min,在此条件下,平均容重为533.17 g/L,沉降速度为7.10 cm/s,初始沉水率为70.33%,10 min沉水率为99.00%。研究结果可为饲料企业生产缓沉性水产膨化饲料提供参考依据。     

Ecotoxicological evaluation of Harbour sediments using marine organisms from different trophic levels

Background, Aim and Scope  The toxicity of contaminated sediments should be evaluated considering the direct exposure of laboratory organisms to whole sediments and the indirect exposure to elutriates or extracts (Tay et al. 1992, Byrne and Halloran 1999, Nendza 2002). The alga Dunaliella tertiolecta is indicated for the use in toxicity bioassays because it is highly sensitive to several xenobiotics. Harpacticoid copepods have been already used for toxicity testing and Tigriopus fulvus is a promising Mediterranean target-species in ecotoxicology (Todaro et al. 2001, Faraponova et al. 2003, Pane et al. 2005a). In this study, the toxicity of sediments collected in harbour sites of the Northeastern Adriatic Sea was evaluated by growth inhibition test with free living and alginate-immobilized Dunaliella tertiolecta and acute toxicity test with nauplii and adult Tigriopus fulvus with the aim of pointing out the importance to utilize model organisms from different trophic levels in sediment ecotoxicology. Methodology  Elutriates and whole sediments were tested on free living and immobilized (Pane et al. 1998) algal cells, and on laboratory reared copepods. Free-living D. tertiolecta were exposed to diluted elutriates in a static, multi-well plate system. Naalginate immobilized D. tertiolecta were placed in polystyrene inserts fitted with polyester mesh bottoms and exposed to a thin layer (2 mm) of whole sediments in multi-well plates (EPS 1992, Pane and Bertino 1999). Toxicity tests with copepods were carried out on Tigriopus fulvus nauplii (elutriates) and adults (whole sediments and elutriates). Same-aged nauplii useful for toxicity tests were obtained by egg sac detaching and consequent hatching stimulation (Pane et al. 2006). Newborn nauplii (I–II stage) were exposed to elutriates in multi-well plates provided with polystyrene inserts. Adult T. fulvus maintained in polystyrene inserts fitted with polyester mesh bottoms were placed in contact with a thin layer (2 mm) of whole sediment placed on multi-well plate bottoms. All end-points were evaluated after 96 h. Results  In general, the effects increased with the increasing of elutriate concentration up to 50%; the stimulation or inhibition of algal growth was statistically significant in comparison to the control. The inhibiting elutriates induced EC₅₀ variations of algal growth ranging from 66.9% to 74.3%. The mortality of T. fulvus nauplii was always < 25% after treatment with 100% elutriates and < 10% after treatment with 50% dilution; no effect was shown up with 25% dilution; therefore LC₅₀ was not calculable. The effect of elutriates was negligible on adult copepods and LC₅₀ values were never calculable; percent mortality always resulted in < 10% after treatment with whole sediments. Discussion  Both experimental systems gave substantially similar results after exposition to whole sediments and elutriates. During the experiment with algal cells, the immobilization in Na-alginate and the employment of inserts which allowed the contact of organisms with sediments and their easy counting were particularly useful. Likewise, the employment of inserts of adequate mesh size in the tests with copepods allowed the contact of organisms with the sediment and made organism handling and counting easy, as well as the evaluation of mortality. The methodology here described and the utilization of the proposed test-species could have an importance also considering that the current trend in ecotoxicological research is towards finding the most appropriate organism for specific areas of concern by using indigenous species (Mariani et al. 2006) and towards the major significance of chronic and reproductive end-points. Conclusions  Based on the above results, it can be stated that the bioassay with Dunaliella tertiolecta could be a good estimation tool for the ecotoxicological assessment of marine sediments. The immobilization of algae in Na-alginate was seen to be useful to evaluate the toxicity of whole sediments; the employment of polystyrene inserts allowed an improvement of the procedures. T. fulvus nauplii and adults, as other harpacticoids such as Tigriopus japonicus (Yoon et al. 2006), satisfy the basic criteria for the employment of a standard species in marine bioassays. To date only pelagic Acartia tonsa are utilized in the standardized procedure to evaluate the risk assessment of chemicals or wastewaters (ISO 1999). As, on the contrary, the exposure of copepods to solid-phase contaminants it is not yet standardized, the employment of polystyrene inserts improved the procedures for T. fulvus too. So, the rapidity and the possibility to solve practical problems could be the main attractive features of this technique (Pane et al. 2005a) when applied to whole sediments. Recommendations and Perspectives  The methodology here developed being also applicable to long term and reproduction tests should be recommended because it provides relevant information in comparison with other frequently applied, standardized biotests with crustaceans (ISO 1999). The procedure has been shown to be easily applicable to selected marine organisms. ESS-Submission Editor: Prof. Dr. Henner Hollert (henner.hollert@bio5.rwth-aachen.de)     

利用QuantStudioTM 3D数字PCR分析转基因玉米MON863含量

QuantStudioTM 3D数字PCR (QuantStudioTM 3D digital PCR,3D-dPCR)是一种基于超高密度亲疏水微孔芯片实现数字PCR分液原理的新型核酸绝对定量平台,在转基因生物定量领域具有极大的应用前景.本研究基于3D-dPCR平台,以转基因玉米(Zea mays)MON863混合样品为例,建立基于单重和双重数字PCR体系的转基因生物(genetically modified organisms,GMOs)含量分析方法.与传统qRT-PCR比较发现,在缺乏样品纯度、纯合度信息的情况下,数字PCR能够较好地排除这些因素的影响,测定准确的量值.研究结果表明,QuantStudioTM 3D数字PCR是一种适用于转基因生物含量分析的精确定量方法,还可反映转基因玉米种子的基因型.本研究基于3D-dPCR建立的转基因玉米MON863单重和双重定量方法为转基因检测提供了新的方法和参考.     

利用微滴数字PCR分析转基因生物外源基因拷贝数

微滴数字PCR(droplet digital PCR,ddPCR)是一种基于泊松分布原理的核酸分子绝对定量技术,在核酸分子的绝对计数/定量领域具有极大的应用潜力。本研究基于ddPCR平台,以转基因水稻(Oryza sativa)T1c-19和转人乳铁蛋白基因基因山羊(Capra hircus)134为例,建立了转基因生物(genetically modified organisms,GMOs)外源基因拷贝数分析方法,并比较了其与传统的实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)和Southern blot方法的准确性。实验数据表明,T1c-19的杀虫晶体蛋白基因(insecticidal crystal protein,Cry1C*)在qRT-PCR和ddPCR测定结果比较一致,约为2拷贝,但已报道的Southern blot分析结果为1拷贝;ddPCR对bar基因的分析结果高于qRT-PCR,分别为2.09拷贝和1.51拷贝。转人乳铁蛋白基因(human lactoferrin,HLF)山羊134在qRT-PCR和ddPCR的分析结果基本一致,均测得含有1拷贝的HLF基因。研究结果表明,微滴数字PCR方法是一种经济、快速和准确的外源基因拷贝数分析新方法,灵敏度和准确性高,将会在拷贝数分析中广泛应用。     

Are survival and reproduction of Enchytraeus albidus (Annelida: Enchytraeidae) at risk by feeding on Bt-maize litter?

Enchytraeids are saprophagous soil organisms, appearing in high abundances and contributing to ecological processes within the soil. For decades they have been used as model species for biological research. In the framework of research on genetically modified plants, however, they have not been considered to date. Following the ISO/DIS guideline, survival and reproduction of Enchytraeus albidus, fed with diets containing Bt-maize (N4640Bt Cry1Ab, DKC5143Bt Cry3Bb1) leaf material were analysed. For comparison, diets with the corresponding untransformed near-isolines (N4640, DKC5143) were examined. Additionally a high quality control diet (oat flakes) was included. Survival and reproduction showed no significant differences between the Cry3Bb1 treatment and the treatment with the untransformed counterpart. For the Cry1Ab treatment survival was significantly higher than for the treatment with the corresponding near-isoline. In contrast, reproduction was significantly lower for the Cry1Ab treatment compared to that for the isoline. For the Cry3Bb1 treatment, no effect was shown on survival or reproduction. For the Cry1Ab variety and its untransformed counterpart, a contrasting result was detected, which is unlikely to be caused by the Bt-protein but rather by differences in other plant components. Overall survival and reproduction were highest for the control.     

温室池水环境与罗非鱼运动性气单胞菌败血症爆发的关系

为阐明温室池水环境对罗非鱼运动性气单胞菌败血症发生的影响,对5年监测所得的发病池的95个水样和无病池的78个水样的水环境因子值进行了统计分析。结果表明,虽然发病池和无病池的水温平均值和pH平均值差异均不显著(p＞0.05),但在18～27 ℃范围内,发病池出现的百分率随水温升高而增加;发病池的溶解氧、总氨氮、分子氨氮、亚硝酸盐氮、有机物化学耗氧量的平均值分别为2.85±1.45、2.48±0.99、0.067±0.048、0.361±0.233、39.56±20.31 mg/L;无病池的溶解氧、总氨氮、分子氨氮、亚硝酸盐氮、有机物化学耗氧量的平均值分别为3.41±1.39、0.94±0.76、0.025±0.025、0.105±0.066、20.62±9.73 mg/L。发病池的溶解氧平均值显著低于无病池(p＜0.05),发病池的总氨氮、分子氨氮、亚硝酸盐氮、有机物化学耗氧量的平均值分别极显著高于无病池(p＜0.01)。根据发病池和无病池出现的百分率随各环境因子变化的趋势可以得出结论：高水温、低溶解氧、高氨氮、高分子氨氮、高亚硝酸盐氮和高有机物化学耗氧量的池水更容易诱发凡隆气单胞菌温和生物型引起的罗非鱼运动性气单胞菌败血症。     

Impact of harvesting and logging slash on nitrogen and carbon dynamics in soils from upland spruce forests in northeastern Ontario

The potential impact of timber harvesting in the boreal forest on aquatic ecosystem water quality and productivity depends in part on the production of nutrients within the soil of the harvested catchment. Nitrogen supplied by organic matter decomposition is of particular interest because of the important role that N plays in biotic processes in surface waters, and in forest nutrition in general. Logging slash quality and input to the forest floor has the potential to influence N availability after harvest on clearcut sites. Net production of organic and inorganic-N and microbial biomass C and N concentrations were determined during a 90-day laboratory incubation at constant temperature and moisture. Incubated soils included F horizon and shallow mineral soil horizons (0-5 cm) from unharvested and full-tree harvested (2 and 12 growing seasons since harvest) boreal forest sites at the Esker Lakes Research Area (ELRA), in northeastern Ontario, Canada. In an ancillary experiment, black spruce foliage was added to unharvested forest floor material after 30 days during a 90-day laboratory incubation to simulate the influence of logging slash from full-tree harvesting on C and N dynamics. Twelve-year old clearcut F horizon material released on average 75 and 5 times more -N and 3 and 2 times as much inorganic-N than soil collected from unharvested and 2-year-old clearcuts, respectively. This increase in -N accumulation during the incubation was accompanied by decreases in both exchangeable -N and microbial biomass C and N levels. Net daily changes in microbial biomass N were significantly related to organic and inorganic-N accumulation or loss within the F horizon. Mineral soil release of inorganic-N was lower than release from the forest floor. Nitrate-nitrogen accumulation was lower, and -N accumulation was higher in mineral soil from unharvested sites when compared to 12-year-old clearcuts. Calculated harvest response ratios indicated that incubated mineral soil from the 12-year-old clearcut sites released significantly greater amounts of -N than 2-year-old clearcuts. Incorporation of black spruce needles into F horizon material reduced the production of organic and inorganic-N and increased microbial biomass N. Laboratory incubations of F horizon and shallow mineral soil from 12-year-old clearcuts suggested that these boreal soils have the capacity for increased inorganic-N production compared to uncut stands several years after harvesting. This has the potential to increase N availability to growing boreal forest plantations and increase N leaching due to greater -N levels in the forest soil.     

双抗体夹心ELISA在转基因生物检测中的应用研究

随着转基因产品种类和数量的不断增加,转基因生物及其产品的安全性也越来越受到关注,各国一直以来也都高度重视转基因生物及产品的管理.作为转基因生物安全评价的第一步,转基因生物的准确检测显得格外重要.该文综述了双抗夹心ELISA方法的原理、技术要点及其在转基因植物及产品检测中的应用,并对该方法在转基因动物检测中的应用前景进行分析,为双抗夹心ELISA方法的合理利用提供依据.     

水产下脚料对肉鸡生长性能、胴体性状及肠道健康的影响

文章旨在评估日粮添加不同水平的水产下脚料对肉鸡生长性能、胴体性状及肠道健康的影响.试验将504只体重一致的1日龄肉鸡随机分为3组,每组6个重复,每个重复28只.对照组饲喂基础日粮,处理组饲喂水产下脚料添加水平为8％和16％的日粮,对照组与处理组日粮的营养水平保持一致,试验为期6周.结果:T1和T2组42?d肉鸡体重、2...     

水产品解冻技术研究进展

在概述常见水产品解冻技术原理的基础上,归纳了不同解冻技术在虾蟹类、软体类和鱼类中的应用,探讨了不同解冻方式对水产品感官、物理、化学及微生物指标的影响。传统解冻技术适用于大部分水产品,是目前最常用的解冻方式,但存在解冻时间长、效率低、微生物污染较严重等缺点。新型解冻技术解冻时间短,能够很好地减少微生物污染,尤其对于一些大型、高价值水产品的解冻颇具优势,但成本和能耗较高。与此相比,传统+新型组合解冻技术能够在一定程度上减少单一技术的负面影响,但同时对于成本和安全性方面仍需要开展进一步研究。