A new plant-based bioassay for aquatic sediments

Background, Goal and Scope  To date, standardised bioassays for the assessment of the ecotoxicological potential in sediments and dredged material use test organisms like bacteria, algae and crustaceae. This paper presents the development and application of a novel sediment contact test (whole sediment) withMyriophyllum aquaticum, a representative of rooted aquatic macrophytes. The aim of the present study is to demonstrate the value of a sediment contact test with rooted macrophytes as a supplement to existing test batteries in order to improve the assessment of sediment toxicity. Methods  The newly developed sediment contact test withMyriophylhim aquaticum was applied to natural whole sediments. For performing the test, whorls ofMyriophyllum aquaticum were directly planted in the native sediment and incubated in the light at 24°C (cf. section results and discussion). The end points of the test were the number of the shoots and the fresh weight of the whole plants. The duckweed growth inhibition test withLemna minor according to ISO/DIS 20079 was performed in pore waters from sediment samples. The results of the sediment contact test withMyriophyllum aquaticum were compared with each other and with those of the aquatic duckweed test. Results and Discussion  A test protocol for the new plant-based sediment contact test using the aquatic plantMyriophyllum aquaticum as an indicator was developed. The best control sediment proved to be the OECD sediment (OECD 207). A test period of 10 days appeared to be sufficient for the test. The increase of biomass and the derived growth rate were found to be the most suitable evaluation parameters. The growth behaviour ofMyriophyllum aquaticum differed depending on the origin of sediments. Therefore, plant-affecting contamination, that is bound in sediments, was indicated. Conclusions  The novel sediment contact test withMyriophyllum aquaticum can indicate phytotoxic effects in sediments. Therefore, it allows a better assessment of the overall-toxicity in whole sediments. Recommendations and Outlook  The sediment contact test withMyriophyllum aquaticum is a valuable tool for the evaluation of the ecotoxicological risk potential of waters and sediments. It should become a complement to a standardised test battery generally used for the assessment of sediment toxicity.     

小麦根尖细胞壁对铝的吸附/解吸特性及其与耐铝性的关系

研究了耐铝性明显差异的2个小麦基因型西矮麦1号(耐性)和辐84系(敏感)根系对铝毒胁迫的反应与根尖细胞壁组分以及细胞壁对铝的吸附和解吸的关系。结果表明,30mol/L.AlCl3可迅速抑制小麦根系伸长,但对辐84系根系伸长的抑制更为明显,且小麦根系相对伸长率随着铝浓度的提高而急剧降低。在30mol/L.AlCl3处理24h后,西矮麦1号根系伸长的抑制率为33.3%,而辐84系根系伸长的抑制率高达70.9%。小麦距根尖0～10.mm根段的铝含量和细胞壁中果胶糖醛酸含量显著高于10～20.mm根段,且前者对铝的累积吸附量明显大于后者;在0～10.mm根段,敏感基因型果胶含量高于耐性基因型,其根尖含铝量及根尖细胞壁对铝的吸附量都要大于后者。采用1.0.mol/L.NH3.H2O对细胞壁预处理2.h降低果胶甲基酯化程度后,耐性和敏感基因型根尖细胞壁对铝的累积吸附量分别降低了17.1%和20.9%,但对铝的累积解吸率没有影响。由此可见,小麦根尖是铝毒的主要位点,细胞壁果胶含量和果胶甲基酯化程度可能是导致不同小麦基因型根尖细胞壁对铝吸附量、铝积累量的差异及其对铝毒胁迫反应的差异的重要原因。     

Changes in photosynthesis and antioxidant metabolism of cotton (Gossypium hirsutum L.) plants in response to manganese stress

ABSTRACT

This study aimed to assess the physiological and biochemical responses of cotton plants to manganese (Mn²⁺) nutrition. Four cotton genotypes (G1 – TMG 47; G2 – FM 975 WS; G3 – TMG 11 WS and G4 – IMA 8405 GLT) were grown in nutrient solution under two Mn²⁺ concentrations (2 and 200 µmol L^?1) for 10 days. No visible symptoms of Mn²⁺ toxicity were observed in the genotypes tested. All genotypes showed a marked increase in leaf chlorophylls, pheophytins, carotenoids, sucrose and total sugars concentration in response to high Mn²⁺ in a nutrient solution. However, the net photosynthetic rate, stomatal conductance, internal carbon dioxide concentration and transpiration decreased in genotypes G1 and G2 growing under 200 µmol L^?1. Antioxidant enzymes such as superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR) activities increased in genotypes G1, G3 and G4. Cotton genotypes showed an increased leaf antioxidant and sugar metabolism as a possible strategy to mitigate oxidative stress. The decrease in the net photosynthetic rate and stomatal conductance; the increased antioxidant enzymes activities (SOD, APX and GR); and the increase in leaf sucrose and total sugar concentration were the main physiological and biochemical responses in cotton plants to Mn²⁺ stress.     

Proton toxicity interferes with the screening of common bean (Phaseolus vulgaris L.) genotypes for aluminium resistance in nutrient solution

Common bean (Phaseolus vulgaris L.) proved to be very sensitive of low pH (4.3), with large genotypic differences in proton sensitivity. Therefore, proton toxicity did not allow the screening of common bean genotypes for aluminium (Al) resistance using the established protocol for maize (0.5 mM CaCl₂, 8 μM H₃BO₃, pH 4.3). Increasing the pH to 4.5, the Ca²⁺ concentration to 5 mM, and addition of 0.5 mM KCl fully prevented proton toxicity in 28 tested genotypes and allowed to identify differences in Al resistance using the inhibition of root elongation by 20 μM Al supply for 36 h as parameter of Al injury. As in maize, Al treatment induced callose formation in root apices of common bean. Aluminium‐induced callose formation well reflected the effect of Ca supply on Al sensitivity as revealed by root‐growth inhibition. Aluminum‐induced callose formation in root apices of 28 bean genotypes differing in Al resistance after 36 h Al treatment was positively correlated to Al‐induced inhibition of root elongation and Al contents in the root apices. However, the relationship was less close than previously reported for maize. Also, after 12 h Al treatment, callose formation and Al contents in root apices did not reflect differences in Al resistance between two contrasting genotypes, indicating a different mode of the expression of Al toxicity and regulation of Al resistance in common bean than in maize.     

噻虫嗪在农田土壤中环境行为的研究进展

噻虫嗪是当前全球销售量最大的新烟碱类农药之一，随着噻虫嗪的广泛使用，噻虫嗪在土壤环境中的迁移转化和归宿问题成为研究热点。本文总结了噻虫嗪对陆生生物（蜜蜂、蚕、蚯蚓、鹌鹑）、水生生物（藻类、大型溞、斑马鱼等）以及土壤微生物的影响，系统综述了噻虫嗪在农田土壤中的残留、降解、吸附解析以及迁移与淋溶等环境行为及主要影响因素，并指出了噻虫嗪在农田土壤中环境行为相关研究存在的不足以及未来研究应该关注的重点和方向，以期为噻虫嗪的科学安全使用以及保障农田土壤生态环境健康提供科学依据。     

Bioassay-directed Fractionation of Organic Extracts of Marine Surface Sediments from the North and Baltic Sea - Part II: Results of the biotest battery and development of a biotest index (8 pp)

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Part I: Determination and identification of organic pollutants Part II: Results of the biotest battery and development of a biotest index

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Preamble. This series of two papers presents the results of an interdisciplinary research project (ISIS) dealing with bioassay-directed fractionation of marine sediment extracts. Part I presents the extraction and fractionation procedure as well as the results of chemical analysis, including non-target analysis of sediments. Part II describes the results of the biotest battery in relation to chemicals possibly causing parts of the observed effects. A biotest index is used to compare the toxicities of the samples.

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AUTHORS / AFFILIATIONS Ninja Reineke (3), Werner Wosniok (4), Dirk Danischewski (1), Heinrich Hühnerfuss (3), Angelika Kinder (5), Arne Sierts-Herrmann (5), Norbert Theobald (2), Hans-Heinrich Vahl (6), Michael Vobach (1), Johannes Westendorf (6) and Hans Steinhart (5).

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(1) Federal Research Centre for Fisheries, Institute for Fishery Ecology, Palmaille 9, 22767 Hamburg, Germany (2) Federal Maritime and Hydrographic Agency, Bernhard-Nochtstr. 78, 20359 Hamburg, Germany (3) University of Hamburg, Institute for Organic Chemistry, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany (4) University of Bremen, Institute of Statistics, Bibliothekstr. 1, 28334 Bremen, Germany (5) University of Hamburg, Institute for Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany (6) University of Hamburg, University Hospital Hamburg-Eppendorf, Department for Toxicology, Vogt-Kölln-Str. 30, 22527 Hamburg, Germany (7) Eurofins Wiertz-Eggert-Jörissen, Stenzelring 14b, 21107 Hamburg, Germany

Goal, Scope and Background

The ecological relevance of contaminants in mixtures is difficult to assess, because of possible interactions and due to lacking toxicity data for many substances present in environmental samples. Marine sediment extracts, which contain a mixture of environmental contaminants in low concentrations, were the object of this study. The extracts were investigated with a set of different biotests in order to identify the compound or the substance class responsible for the toxicity. For this goal, a combination of biotests, biotest-directed fractionation and chemical analysis has been applied. Further on, a strategy for the development of a biotest index to describe the toxicity of the fractions without a prior ranking of the test results is proposed. This article (Part II) focuses on the biological results of the approach.

Methods

The toxicological potential of organic extracts of sediments from the North Sea and the Baltic Sea was analyzed in a bioassay-directed fractionation procedure with a set of biotests: luciferase reporter gene assays on hormone receptor and Ah receptor, arabinose resistance test, fish embryo test (Danio rerio), comet assay, acetylcholinesterase inhibition test, heat-shock protein 70 induction, oxidative stress and luminescence inhibition test (Vibrio fischeri). The test results provided the basis for the calculation of a biotest index by factor analysis to compare the toxicity of the samples and fractions.

Results and Discussion

Results of 11 biotests on different fractionation levels of the samples were described and discussed with regard to the occurrence of contaminants and their toxic potentials. Polychlorinated biphenyls, polycyclic aromatic hydrocarbons, quinones, brominated indoles and brominated phenols were in the focus of interest. A biotest index was constructed to compare the toxic responses in the samples and to group the biotest results.

Conclusion

The procedure presented in this study is well suited for bioassay-directed fractionation of marine sediment extracts. However, in relatively low contaminated samples, high enrichment factors and sufficient fractionation is necessary to allow identification of low concentrations of contaminants which is required to link effects and possible causes. In the present case, the relation between substances and effects was difficult to uncover due to relatively low concentrations of pollutants compared to the biogenic matrix and to the remaining complexity of the fractions. The results, with respect to the brominated phenols and indoles in the samples, highlight the successful use of bioassay directed fractionation in the case of high concentrations and high toxicity.

Recommendation and Outlook

In general, it has been shown that a marine risk assessment requires focusing on the input of diffuse sources and taking into account the fact of mixture toxicity. Effects resulting from biogenic substances will make the assessment of the influence of anthropogenic substances even more difficult.     

外源低分子量有机酸对碳酸钙预处理的酸性土壤中活性铝钙镁影响的研究

通过在所研究的第四纪红黏土发育的红壤中混入CaCO3,研究在pH缓冲体系中外加低分子量有机酸对土壤中Al、Ca和Mg的影响。结果表明:无论加CaCO3与否,在pH 4.5的条件下外源草酸、柠檬酸、苹果酸的加入均使土壤可溶性Al显著提高,交换性Al显著下降和交换性Ca显著升高;加入CaCO3的情况下,3种有机酸处理的交换性Mg均显著提高。3种有机酸促进Al溶解能力的大小顺序为:柠檬酸>草酸>苹果酸,这一结果与有机酸和Al形成络合物的稳定常数大小一致。另一方面,3种有机酸处理下,CaCO3预处理均引起可溶性Al的显著升高和交换性Al的下降。双因素方差分析表明,有机酸通过络合作用或沉淀作用对可溶性和交换性Al、Ca和Mg均具有绝对的影响优势,CaCO3仅对可溶性和交换性Al、交换性Ca有显著影响,由于实验中pH缓冲体系的控制,这种影响主要通过Al与Ca、Mg的竞争交换作用实现。总体来说,外源低分子量有机酸的加入使土壤活性Al显著升高,活性Ca、Mg略有升高,有机酸在酸性土壤中的作用需从有机酸溶解阳离子的角度进一步评价。     

铜污染对高等植物的生理毒害作用研究

本文综述了当前国内外Cu污染对高等植物生理毒害的研究近况,探讨了Cu过量对植物光合作用、细胞结构、细胞分裂、酶学系统和其他营养元素的吸收等的影响,并探讨了该方面研究存在的不足及其展望。     

恩诺沙星与磺胺二甲嘧啶联合毒性探究

本研究旨在探究恩诺沙星与磺胺二甲嘧啶的联合毒性。选取SD大鼠为试验对象,将其分成6组,每组12只,分别为高剂量联合用药组（500 mg/kg体重）、中剂量联合用药组（250 mg/kg体重）、低剂量联合用药组（50 mg/kg体重）、恩诺沙星单药组（250 mg/kg体重）、磺胺二甲嘧啶单药组（250 mg/kg体重）及对照组（等量0.5%羧甲基纤维素钠溶液）,配制相应的药物进行灌胃,最后一次给药1 d后对大鼠进行称重、麻醉、心脏采血并剖检取肝脏组织,然后对大鼠增重率、血常规指标、血清生化指标及肝脏病理学变化几个方面进行分析,并利用SPSS 22.0软件评估2种药物的联合作用效果。结果显示,在雌性大鼠中,高、中剂量联合用药组使大鼠增重率显著下降（P<0.05）,雄性大鼠增重率无显著变化（P>0.05）;雄性大鼠给药组白细胞数量均显著增加（P<0.05）,高剂量联合用药组中性粒细胞和淋巴细胞占比分别表现为显著升高及下降（P<0.05）;高、中剂量联合用药组大鼠外周血中谷草转氨酶含量显著增加（P<0.05）,且高、中剂量联合用药组肝脏病理切片视野可见不同程度的炎性细胞浸润,并伴有不同程度的损伤。本研究结果表明,恩诺沙星和磺胺二甲嘧啶联用可使毒性增加,对大鼠的免疫系统状态有一定的影响,且能造成一定程度的肝脏损伤,剂量越高影响越大。本研究为恩诺沙星和磺胺二甲嘧啶联合用药机制的研究提供数据支持,并为二者的临床应用提供参考,提示新的食品安全评估应考虑药物联合暴露带来的影响。     

芩术安胎散的毒性和临床安全性试验研究

试验通过对芩术安胎散的急性毒性、亚慢性毒性和临床安全性进行研究,为芩术安胎散对家猫先兆性流产的预防与治疗提供参考。急性毒性试验：制备芩术安胎散药液,取6周龄健康昆明小白鼠60只,随机分为5组,每组12只,第1~4组的给药剂量分别为6 000、4 800、3 840和3 072 mg/kg,对照组给予等量纯净水,10 d内观察有无中毒和死亡,计算半数致死量（LD₅₀）;另取6周龄健康昆明小白鼠20只,随机分为2组：试验组给予2.0 g/mL芩术安胎散药液,18 h内灌服3次,每次0.8 mL,对照组给予等量纯净水,给药后饲养7 d,计算最大耐受量（MTD）。亚慢性毒性试验：24只7周龄SD雌性大鼠随机均分为高、中、低剂量组和对照组,在30 d内,每日分别给予4 800、2 400和1 200 mg/kg芩术安胎散,对照组以等量纯净水进行灌胃,每日观察和记录各组大鼠的精神状态、有无中毒症状和死亡;第31天对各组大鼠称重、采血,进行血液学检测,剖检各组大鼠,观察主要脏器有无病变并制作病理切片。临床安全性试验：选取2~5岁健康雌性家猫20只,适应性饲养10 d,随机分组,每组5只,分别为低剂量组（1倍临床推荐剂量：1.15 g/kg）、中剂量组（3倍临床推荐剂量：3.45 g/kg）、高剂量组（5倍临床推荐剂量：5.75 g/kg）及空白对照组,将药物置于胶囊内,口服给药,空白对照组给予空胶囊,每日1次,连续给药7 d,每日观察各组家猫食欲、精神状态及排便情况,于第8天对各组家猫进行静脉采血,检测血常规和血液生化指标。结果显示,急性毒性试验无小鼠死亡,LD₅₀>6 000 mg/kg;小鼠对芩术安胎散的最大耐受量为240 g/kg,表明该受试药物无明显毒性。在亚慢性毒性试验中,各组大鼠的生长发育情况、血常规指标、脏器系数与对照组相比均无显著差异（P>0.05）;高、中剂量组与低剂量组、对照组相比血清总胆固醇含量显著下调（P<0.05）,除此之外的生化指标均无显著差异（P>0.05）。病理剖检和组织切片观察结果显示,高剂量组大鼠主要组织器官与对照组相比无明显异常。在临床安全性试验中,不同剂量组家猫精神状态、被毛光泽度、粪便情况均正常,血液学指标与对照组相比差异均不显著（P>0.05）。本试验结果表明,中药芩术安胎散无明显毒性,家猫按临床推荐剂量使用是安全的。