Persistence of malachite green and leucomalachite green in perch (Lateolabrax japonicus)

The persistence of malachite green (MG), and its metabolite leucomalachite green (LMG), in fish tissues is still unclear, leading to many trade disputes. In this research, we established and evaluated an HPLC method that could detect MG and LMG simultaneously, and then investigated the persistence of these two toxins in the tissues of juvenile perch (Lateolabrax japonicus) post sub-chronic MG exposure at 1.0 mg/L. Exposure lasted for 2 h everyday and was repeated six times. The perch were then placed in MG-free seawater for 100 d to eliminate the toxins. Results show that MG accumulated in the tissues, including the gills, liver, muscle, blood and viscera, and then was metabolized rapidly to LMG. The concentrations of these two toxins increased significantly with the accumulation process. In general, the highest concentrations of MG and LMG in all tissue exceeded 1 000 μg/kg, except for MG in the muscle. The order of accumulation levels (highest to lowest) of MG was gill>blood>liver>viscera>muscle, while that of LMG was liver>blood>gill>viscera>muscle. High levels of MG or LMG could persist for several hours but decreased rapidly during the elimination process. The concentration of LMG was much higher than that of MG during the experiment, especially in the gill, liver and blood. Therefore, the three tissues play important roles in toxin accumulation, biotransformation, and elimination. Although the MG and LMG concentrations in muscle were much lower than in other tissues, the content still exceeded the European minimum required performance limit (MRPL), even after 2 400 h (100 d) of elimination. This demonstrates that it is extremely difficult to eliminate MG and LMG from tissues of perch, and therefore use of these toxins is of concern to public health.     

高效液相色谱法检测水产品中的孔雀石绿和结晶紫

样品中的孔雀石绿、结晶紫经试剂盒提取,浓缩后用经固相萃取柱净化、硼氢化钾还原、反向色谱柱分离,使用荧光检测器检测,孔雀石绿、结晶紫的加标回收率在76.1%～92.5%之间,相对标准偏差1.9%～5.8%,检出限均为0.5μg/kg。结果表明:该方法检测孔雀石绿(MG)、结晶紫(GV)处理简单,灵敏度高,节省时间,可用于大量样品的快速分析。     

一种基于移动窗口的城市绿度遥感度量方法

针对城市绿度空间分布特征难以描述且合理性难以度量的问题,在基于多光谱遥感数据和激光雷达数据（LiDAR）获取高精度城市建筑物与绿地信息的基础上,本文提出一种基于移动窗口的城市绿度遥感度量方法。其通过遍历研究区像元,建立了移动窗口内的城市绿地面积与固定窗口面积之间的关系,计算了中心像元的绿度指数,并与传统的格网法、缓冲区法进行了比较。研究结果表明：该方法建立的绿度指数,能很好地反映城市居民在区域内接触城市绿地的概率及其空间分布特征,与格网法和缓冲区法相比,有效避免了格网法产生的边缘效应,并解决了缓冲区法空间不连续性的问题,实现了区域范围内任意点的城市绿度度量。该方法为城市绿地度量、城市规划、管理提供了一种新视角。     

大连市城区绿地时空特征的热环境效应研究

本文选择大连市沙河口和西岗区为研究区,以Landsat ETM、SPOT等数据为基础,通过遥感解译和反演得到1999年和2013年研究区内绿地及地表温度空间分布及演变特征,结合温度-植被指数（TVX）空间分析法测度城区发展过程中绿色空间减退所引起的热环境效应,以提出更有针对性的城市气候变化适应性规划。研究结果表明：（1）1999-2013年,研究区绿地面积减少43.8%,绿地斑块增加42.9%,城市绿地呈现面积减少、空间分布碎化的演变特征。其中,森林绿地、公园绿地和街区绿地转化为建设用地的贡献率分别为43.8%、42.9%和13.3%,街区绿地因原始面积小且转化率高,被建设用地侵占最为严重。（2）1999-2013年,研究区地表温度最高值上升5.2 K,地表温差增加1.7 K,地温聚集区间由301.6 ~304.7 K上升至306.5~310.9 K,热环境效应加剧。（3）TVX模型追踪结果显示,快速城市化过程中研究区内城市绿地转化为建设用地引起植被覆盖度下降和地表温度上升,其中街区绿地被侵占造成的地表热环境效应大于公园绿地和森林绿地。     

高效液相色谱法测定罗非鱼中孔雀石绿残留量的不确定度

依据JJF10591—2012《测量不确定度评定与表示》,分析高效液相色谱法测定罗非鱼中孔雀石绿残留量不确定度的来源,从影响不确定度的标准溶液配制us、样品前处理up和仪器测量uu等3个主要因素,对不确定度各个分量进行计算和合成,得出罗非鱼中孔雀石绿的扩展不确定度U=088（k=2）。     

Spatio-temporal variability of small fishes related with environmental factors in a typical domestic tap water lake,Eastern China

The knowledge of prey small fish stock, distribution and abundance is necessary to guide stocking of piscivorous fish for the biomanipulation in domestic tap water lakes. This study describes the current status of small fish community in Lake Kuilei (China), and examines the spatial and seasonal variations of the community in relation to key environmental factors. Based on submerged macrophyte cover and water depth, the lake was divided into five major habitats: (1) macrophyte covered shallow habitat of water depth < 2.00 m, (2) uncovered or less-covered shallow habitat (2.00 m–3.50 m), (3) uncovered medium shallow habitat (3.50 m–5.00 m), (4) uncovered medium deep habitat (5.00 m–6.50 m) and (5) uncovered deep habitat (6.50 m–8.50 m). The abundance and composition of small fish were monitored by benthic fykenet sampling from April 2013 to January 2014. A total of 2881 individuals belonging to 5 families and 21 species were collected. Based on their abundance (accounted for 88.96% of the total) and occurrence (more than 33.33%), Acheilognathus chankaensis, Acheilognathus macropterus, Microphysogobio microstomus, Pseudorasbora parva and Rhinogobius giurinus were recognized as dominant small fish species. The results of correlation analysis identified that species richness ( Sr ), Shannon-Wiener diversity index ( H′ ) and Margalef′s richness index ( D ) were significantly negatively correlated with water depth, but positively correlated with biomass of submerged macrophytes.Redundancy analysis (RDA) revealed that the spatial distributions of most small fishes were negatively associated with water depth. The details of these findings are beneficial to understanding the adaptation of the small fishes in degraded environments, and to developing suitable biomanipulation strategies for the management of fish resources and water quality in the lakes along the lower reach of the Changjiang (Yangtze) River basin.

     

北京城市绿化的时空动态分析

城市绿化覆盖是城市生态系统的重要组成部分,合理的绿化率和绿化布局可以改善城市环境,提高城市人居适宜性。研究中将2005、2010、2015年北京市土地利用（LUC）数据中的城市居民用地作为城区范围,应用Landsat 5、GF-1影像数据和MODIS产品,利用支持向量机的监督分类方法,提取了2005、2010、2015年的北京市城市绿化覆盖数据,并获取了同期的植被指数（NDVI）数据;继而以城市绿化覆盖率、绿化覆盖均匀度和植被指数为评价指标,在公里栅格和行政区2个尺度上探讨了北京市城市绿化覆盖的空间分布格局和时间变化动态特征。研究表明：① 3个指标在空间和时间2个维度、区县和栅格2个尺度上都表现一致。这反映北京市过去10年中,在绿化面积增加的同时,绿化的空间布局得到优化改善,绿化的质量得到提高。② 2005-2015年,北京市城市绿化覆盖面积由518.93 km² 增加到1405.54 km²,绿化覆盖率由39.9%增加到49.13%,绿化覆盖均匀度由0.598增加到0.653,植被指数由0.42增加至0.5。③ 北京市城市绿化建设存在明显的时空差异。中心城区绿化建设缓慢,成效不明显;重大绿化建设成果主要集中在城市边缘地区和远郊区县。城市绿化改善过程主要发生在2005-2010年。     

基于网格的街区尺度城市绿度度量方法

本文提出一种基于网格的街区尺度城市绿度度量方法。其以900 m×900 m的网格单元作为以城市主干道划分所形成街区尺度的表征,根据网格单元内植被覆盖度与植被构成之间的关系构建绿度指标。通过与传统的基于植被面积占比的格网法所计算的绿度指数进行比较。发现本文方法建立的绿度指标能在街区尺度上提供更加丰富的城市绿度信息,有效地解决了传统方法难以反映的植被覆盖度近似区域因其内部植被构成不同所带来的绿度差异问题。为城市规划、设计以及管理提供了一种新视角。基于该绿度指标,利用Landsat TM/OLI 2009、2015和2019年不同季节的遥感数据分析北京市城市绿化的时空变化特征。结果表明：北京市主城区城市绿化的面积和质量在2009—2019年显著提升,且秋季最为明显。绿化的变化在四环内外呈现出不同的模式。四环内主要为绿化面积的增加,四环外则在绿化面积和绿化质量上都有明显的增长。     

Distribution and expression in vitro and in vivo of DNA vaccine against lymphocystis disease virus in Japanese flounder (Paralichthys olivaceus)

Lymphocystis disease, caused by the lymphocystis disease virus (LCDV), is a significant worldwide problem in fish industry causing substantial economic losses. In this study, we aimed to develop the DNA vaccine against LCDV, using DNA vaccination technology. We evaluated plasmid pEGFP-N2-LCDV1.3 kb as a DNA vaccine candidate. The plasmid DNA was transiently expressed after liposome transfection into the eukaryotic COS 7 cell line. The distribution and expression of the DNA vaccine (pEGFP-N2-LCDV1.3kb) were also analyzed in tissues of the vaccinated Japanese flounder by PCR, RT-PCR and fluorescent microscopy. Results from PCR analysis indicated that the vaccine-containing plasmids were distributed in injected muscle, the muscle opposite the injection site, the hind intestine, gill, spleen, head, kidney and liver, 6 and 25 days after vaccination. The vaccine plasmids disappeared 100 d post-vaccination. Fluorescent microscopy revealed green fluorescence in the injected muscle, the muscle opposite the injection site, the hind intestine, gill, spleen, head, kidney and liver of fish 48 h post-vaccination, green fluorescence did not appear in the control treated tissue. Green fluorescence became weak at 60 days post-vaccination. RT-PCR analysis indicated that the mcp gene was expressed in all tested tissues of vaccinated fish 6–50 days post-vaccination. These results demonstrate that the antigen encoded by the DNA vaccine is distributed and expressed in all of the tissues analyzed in the vaccinated fish. The antigen would therefore potentially initiate a specific immune response. the plasmid DNA was injected into Japanese flounder (Paralichthys olivaceus) intramuscularly and antibodies against LCDV were evaluated. The results indicate that the plasmid encoded DNA vaccine could induce an immune response to LCDV and would therefore offer immune protection against LCD. Further studies are required for the development and application of this promising DNA vaccine.     

A New Molecular Label Applied to the Study of the Yellow Sea Green Tide

From 2007 to 2017, large-scale green tides occurred every year in the Yellow Sea of China, and Ulva prolifera was the main species leading to the green tide. In this study, we used the Polymerase chain reaction and 3' Rapid-amplification of c DNA ends technique to amplify the nr DNA-LSU and IGS sequences in U. prolifera and one species of Blidingia. These techniques showed 3259 bp of nrD NA-LSU and 3388 bp of IGS in U. prolifera and 3282 bp nr DNA-LSU and 3059 bp IGS in Blidingia sp. At the same time, tandem repeats, short dyads, palindromic and multiple simple repeat sequences in the IGS sequence were found by analyzing the structure of the IGS sequence in U. prolifera and Blidingia sp. G + C contents of the IGS sequence in U. prolifera and Blidingia sp. were 52.42% and 53.09%, respectively. We divided the U. prolifera into two types according to the morphological characteristics. Although the specimens of U. prolifera from the Qingdao coastal area, Jiangsu coastal area and the Yellow Sea have different morphologies, their ITS and IGS sequences are almost identical. Therefore, the main species of the green tides in the Qingdao coastal area, Jiangsu coastal area and the Yellow Sea are the same and have the same origin.     

Analysis of IL-2-1ike factor in lymphocyte culture supernatant of olive flounder, Paralichthys oliveaceus

To study immune mechanism of fish lymphocyte we performed a proliferation assay and ELISA using monoclonal antibody against human IL-2. The result showed that an interleukin-2 (IL-2)-like factor was detected in the supernatant of plant haemoglutinin (PHA)-stimulated lymphocyte culture from peripheral blood, spleen and head kidney of olive flounder,Paralichthys olivaceus. The quantities of IL-2-like factor in the supernatant from different lymphoid tissues were quite different. The IL-2 like factor in the supernatant from cultured head kidney lymphocytes was much higher than those of peripheral blood lymphocytes and spleen lymphocytes (P<0.01). The IL-2 activity was found in either mouse thymocyte proliferation assay or flounder head kidney lymphocyte proliferation assay and shown to have obvious enhancing effect on proliferation of the above two types of cell. The recombinant human IL-2, (rhIL-2) was able to stimulate flounder thymocyte proliferation and used to detect the IL-2 receptor (IL-2R) on the surface of flounder lymphocyte. The cross-reaction between the lymphocytes of flounder peripheral blood and CD25(IL-2R) was detected with flow cytometry and shown that the percentage of CD25-positive cell in peripheral blood was 7.74±0.67%. This work was supported by National “973” Project G1999012003, G19999012006.     

Effects of Dietary Protein Level on Growth and Utilization of Protein and Energy by Juvenile Mangrove Red Snapper (Lutjanus argentimaculatus )

1Introduction The mangrove red snapper,Lutjanus argentimacu-latus(Forsskal,1775)is a carnivorous,warm-watereuryhaline fish that is considerably cultured in South-east Asia,Southern China and the Middle East(Le-ung et al.,1999;Estudillo et at.,2000;Ng et al.,2000;Catacutan et al.,2001).In Pakistan,it isknown for its good quality meat and also for its highconsumption rate.Although it fetches a premiumprice at local markets(Anonymous,2002),the in-creasing demand has generated interest towar…     

Preparation by enzymolysis and bioactivity of iron complex of fish protein hydrolysate （Fe-FPH）from low value fish

Preparation of Fe^2＋ chelate of fish protein hydrolysate （Fe-FPH） obtained from low value fish proteins was introduced and its bioactivity was studied by compound enzymolysis. The optimum conditions for hydrolysate chelating Fe^2＋ are DH （degree of hydrolysis） at 5%, pH 7.0, 20℃ and 15 min chelating time for FM （material not being defatted）. Four types of Fe-FPH including CA （deposit after chelating）, CB （deposit in 50% of absolute ethanol solution）, CC （suspended deposit in 80% of absolute ethanol solution）, and CD （bottom deposit in 80% of absolute ethanol solution） were fractionated with absolute ethanol from FM. Structural analysis through infra-red spectrum revealed that Fe^2＋ was combined strongly with amino-group and carboxyl-group in each chelate and each Fe^2＋ could form two five-member ring structures. All of the four chelates were shown more significant antioxidative activity and can be used as natural hydrophobic and hydrophilic antioxidant. Among all the chelates, the CB possesses the most effective antioxidative activity at 92% as high as that of a-tocopherol. Among all Fe-FPHs, only CD showed the most effective antibacterial activity against Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Bacillus subtilis and can be used as natural antibacterial. It provides a more effective way for utilization of low value fish proteins and key information of Fe-FPH as additive in food industry.     

基于城市园林树木景观三维模拟的绿视率估算方法

城市生态景观功能主要是绿色植物对人的视觉作用。绿视率被认为是一种比较好的衡量绿色空间视觉感受的描述因子。目前绿视率的估算主要基于静态的图像或者街景数据,而绿视率是一种动态的量,不同视点有不同的值,且植物是生长变化的。因此,本文提出一种基于数据和模型综合驱动的园林树木三维模拟景观的绿视率计算方法,其利用虚拟地理环境、虚拟植物等技术,通过道路、建筑物等硬质景观数据和树木模型驱动,建立城市园林树木三维景观;根据视觉成像原理,构建虚拟相机,模拟不同视点园林景观的视觉图像,然后识别表征植被信息的像素,从而计算绿视率。本文研发了园林树木景观三维模拟及绿视率估算原型系统,并以城市道路树木景观为例,模拟分析了机动车道中车辆乘客可获得的绿视率,与街景图像提取的绿视率值相近。该方法和系统可用于包含时间变量的不同生长阶段园林树木景观的绿视率评估,支持交互设置不同的视点参数,估算城市任意位置和任意方向的绿视率,评价过去、现在和未来的园林景观绿化质量,以人的视觉感知角度为城市绿地规划提供参考。     

Long-term monitoring of the individual self-feeding behavior of rainbow trout Oncorhynchus mykiss

Self-feeding device is extensively used in aquaculture farms, but for salmonids the individual feeding behavior has seldom been continuously observed. In this article, the individual self-feeding behavior of 10 rainbow trout was continuously monitored with a PIT tag record for 50 days with three replicates. The fish fell into three categories according to their feeding behavior, i.e. high triggering fish (trigger behavior more than 25% of the group, HT), low triggering fish (1%–25%, LT) and zero triggering fish (less than 1%). The results showed that in a group of 10 individual 1–2 HT fish accounted for most of the self-feeding behavior (78.19%–89.14%), which was far more than they could consume. The trigger frequency of the fish was significantly correlated with the initial body weight (P <0.01), however, no significant difference in growth rate among the HT, LT, and ZT fish was observed (P >0.05). Cosinor analysis showed that the two HT fish in the same group had similar acrophase. Though some of the HT fish could be active for 50 d, there were also HT fish decreased triggering behavior around 40 d and the high trigger status was then replaced by other fish, which was first discovered in salimonds. Interestingly, the growth of the group was not affected by the alternation triggering fish. These results provide evidence that in the self-feeding system the HT fish didn’t gain much advantage by their frequent self-feeding behavior, and high trigger status of the HT fish is not only an individual character but also driven by the demand of the group. In the self-feeding system, the critical individual should be closely monitored.

     

“地球村”的绿色与绿色文化

本文扼要介绍近期出现的绿色变革浪潮,提出绿色文化的概念,阐述绿色的价值、绿色文化的一般原则、绿色文化与其它文化的关系,初探绿色社区建设的构想。展示绿色未来,要正视以往由于人类对地理环境的偏见和人类中心主义所招致生态危机,重新调整和确立良好的人地关系。其对地球村的发展有深远意义。     

Applicability of <Emphasis Type="Italic">Perinereis aibuhitensis</Emphasis> Grube for fish waste removal from fish cages in Sanggou Bay,P. R. China

The present study investigated the applicability of integrated polychaete-fish culture for fish waste removal to offset negative impact induced by organic benthic enrichment. A field study demonstrated that deposition rate was significantly higher underneath the fish farm than that in control area. The material settling under the farm was characterized by a high amount of fish feces (45%) and uneaten feed (27%). Both feeding rate (FR) and apparent digestibility rate (ADR) increased with decreasing body weight, as was indicated by significantly a higher rate observed for the groups containing smaller individuals in a lab study. The nutrient in fresh deposited material (De) was higher than that in sediments collected under the farm (Se), resulting in lower feces production but higher apparent digestibility rate for the De group as feeding rate was similar. Consequently, higher nutrient removal efficiency was observed in the De group. A mass balance approach indicated that approximately 400–500 individuals m?2 is required for removing all waste materials deposited underneath the fish farm, whereas abundance can be lower (about 300–350 individuals m^?2) when only the fish waste needs to be removed. The results showed that a significant amount of waste had been accumulated in the fish cages in Sanggou Bay. The integration of fish with P. aibuhitensis seems promising for preventing organic pollution in the sediment and therefore is an effective strategy for mitigating negative effect of fish farms. Thus such integration can become a new IMTA (integrated multi-trophic aquaculture) model in Sanggou Bay.     

Dynamic Diurnal Changes in Green Algae Biomass in the Southern Yellow Sea Based on GOCI Images

Macroalgae blooms of Ulva prolifera have occurred every summer in the southern Yellow Sea since 2007, inducing severe ecological problems and huge economic losses. Genesis and secular movement of green algae blooms have been well monitored by using remote sensing and other methods. In this study, green algae were detected and traced by using Geostationary Ocean Color Imager(GOCI), and a novel biomass estimation model was developed from the relationship between biomass measurements and previously published satellite-derived biomass indexes. The results show that the green algae biomass can be determined most accurately with the biomass index of green algae for GOCI(BIGAG), which is calculated from the Rsurf data that had been atmospherically corrected by ENVI/QUAC method. For the first time, dynamic changes in green algae biomass were studied over an hourly scale. Short-term biomass changes were highly influenced by Photosynthetically Available Radiation(PAR) and tidal phases, but less by sea surface temperature variations on a daily timescale. A new parameter of biomass changes(PBC), calculated by the ratio of the biomass growth rate to movement velocity, could provide an effective way to assess and forecast green tide in the southern Yellow Sea and similar areas.     

Effect of dietary olaquindox on the growth of large yellow croaker (Pseudosciaena crocea R.) and the distribution of its residues in fish tissues

Olaquindox(OLA), one of quinoxaline-N, N-dioxides, has been put under ban. However it was used as a medicinal feed additive early; it promotes the growth of livestock and prevents them from dysentery and bacterial enteritis. In this study, we evaluated the effect of dietary OLA on the growth of large yellow croaker(Pseudosciaena crocea R.) and the histological distribution of OLA and its metabolite 3-methyl-quinoxaline-2-carboxylic acid(MQCA) in fish tissues. Four diets containing 0(control), 42.5, 89.5 and 277.2 mg kg-1 OLA, respectively, were formulated and tested, 3 cages(1.0 m × 1.0 m × 1.5 m) each diet and 100 juveniles(9.75 ± 0.35 g) each cage. The fish were fed to satiation twice a day at 05:00 am and 17:00 pm for 8 weeks. The survival rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet containing 0 and 277.2 mg kg-1 OLA(P 0.05), while the weight gain rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet without OLA(control)(P0.05), but similar to that of fish fed the diet with 277.2 mg kg-1 OLA. Fish fed the diet with 277.2 mg kg-1 OLA had the highest content of OLA and MQCA in liver(3.44 and 0.39 mg kg-1, respectively), skin(0.46 and 0.09 mg kg-1, respectively) and muscle(0.24 and 0.06 mg kg-1, respectively). In average, fish fed the diet containing OLA had the highest content of OLA and MQCA in liver which was followed by skin and muscle(P 0.05), whereas OLA and MQCA were not detectable in control. Our findings demonstrated that OLA and MQCA accumulated in large yellow croaker when it was fed with the diet containing OLA, thus imposing a potential safety risk to human health.