Predicting the aquatic risk of realistic pesticide mixtures to species assemblages in Portuguese river basins

Although pesticide regulatory tools are mainly based on individual substances, aquatic ecosystems are usually exposed to multiple pesticides from their use on the variety of crops within the catchment of a river. This study estimated the impact of measured pesticide mixtures in surface waters from 2002 and 2008 within three important Portuguese river basins (‘Mondego’, ‘Sado’ and ‘Tejo’) on primary producers, arthropods and fish by toxic pressure calculation. Species sensitivity distributions (SSDs), in combination with mixture toxicity models, were applied. Considering the differences in the responses of the taxonomic groups as well as in the pesticide exposures that these organisms experience, variable acute multi-substance potentially affected fractions (msPAFs) were obtained. The median msPAF for primary producers and arthropods in surface waters of all river basins exceeded 5%, the cut-off value used in the prospective SSD approach for deriving individual environmental quality standards. A ranking procedure identified various photosystem II inhibiting herbicides, with oxadiazon having the relatively largest toxic effects on primary producers, while the organophosphorus insecticides, chlorfenvinphos and chlorpyrifos, and the organochloride endosulfan had the largest effects on arthropods and fish, respectively. These results ensure compliance with European legislation with regard to ecological risk assessment and management of pesticides in surface waters.     

Predicting the aquatic risk of realistic pesticide mixtures to species assemblages in Portuguese river basins

Although pesticide regulatory tools are mainly based on individual substances, aquatic ecosystems are usually exposed to multiple pesticides from their use on the variety of crops within the catchment of a river. This study estimated the impact of measured pesticide mixtures in surface waters from 2002 and 2008 within three important Portuguese river basins(‘Mondego', ‘Sado' and ‘Tejo') on primary producers, arthropods and fish by toxic pressure calculation. Species sensitivity distributions(SSDs), in combination with mixture toxicity models, were applied. Considering the differences in the responses of the taxonomic groups as well as in the pesticide exposures that these organisms experience, variable acute multi-substance potentially affected fractions(ms PAFs) were obtained. The median msP AF for primary producers and arthropods in surface waters of all river basins exceeded 5%, the cut-off value used in the prospective SSD approach for deriving individual environmental quality standards. A ranking procedure identified various photosystem II inhibiting herbicides, with oxadiazon having the relatively largest toxic effects on primary producers, while the organophosphorus insecticides, chlorfenvinphos and chlorpyrifos, and the organochloride endosulfan had the largest effects on arthropods and fish, respectively. These results ensure compliance with European legislation with regard to ecological risk assessment and management of pesticides in surface waters.     

Biodegradation of organochlorine pesticide endosulfan by bacterial strain Alcaligenes faecalis JBW4

The recently discovered endosulfan-degrading bacterial strain Alcaligenesfaecalis JBW4 was isolated from activated sludge. This strain is able to use endosulfan as a carbon and energy source. The optimal conditions for the growth of strain JBW4 and for biodegradation by this strain were identified, and the metabolic products of endosulfan degradation were studied in detail. The maximum level of endosulfan biodegradation by strain JBW4 was obtained using broth at an initial pH of 7.0, an incubation temperature of 40℃ and an endosulfan concentration of I00 mg/L. The concentration of endosulfan was determined by gas chromatography. Strain JBW4 was able to degrade 87.5% of α-endosulfan and 83.9% of β-endosulfan within 5 days. These degradation rates are much higher than the previously reported bacterial strains. Endosulfan diol and endosulfan lactone were the major metabolites detected by gas chromatography-mass spectrometry; endosulfan sulfate, which is a persistent and toxic metabolite, was not detected. These results suggested that A. faecalis JBW4 degrades endosulfan via a non-oxidative pathway. The biodegradation of endosulfan by A. faecalis is reported for the first time. Additionally, the present study indicates that strain JBW4 may have potential for the biodegradation of endosulfan residues.     

Antioxidant and modulatory role of Chlorophytum borivilianum against arsenic induced testicular impairment

Arsenic has a suppressive influence on spermatogenesis and induces impairment in male reproductive system due to oxidative stress. The present study was aimed to test the arsenic induced toxicity and protection by Chlorophytum borivilianum. The effect of sodium arsenite (4 mg/(kg body weight (bw). day)) via double distilled water without or with C. borivilianum (800 mg/(kg bw. day)) was evaluated in Swiss albino mice for 30 days. The radical scavenging activity of the aqueous C. borivilianum root extract was measured using DPPH (1,1-diphenyl-2-picryl hydrayzyl) radical. Qualitative assessment of various cell types in the testis, sperm count and motility, testicular activity of lipid peroxidation (LPO), reduced glutathione (GSH), acid and alkaline phosphatase, cholesterol and serum testosterone were monitored. Arsenic treatment showed a significant increase in LPO, acid and alkaline phosphatase, cholesterol and decrease in sperm count, sperm motility, GSH and serum testosterone. Combined treatment showed significant decrease in LPO, acid and alkaline phosphatase, cholesterol and elevation in sperm count, sperm motility, GSH and serum testosterone. Testicular histopathology showed that C. borivilianum had reduced degeneration of germ cell in the seminiferous tubules and loss of sperms induced by arsenic intoxication. The results thus led us to conclude that administration of C. borivilianum root extract is found to be protective against arsenic induced toxicity.     

Endosulfan in the Chinese environment: monitoring and modeling

This paper reviews the usage and emissions of endosulfan, the newest member of the persistent organic pollutants (POPs), in China, and its fate and behavior in Chinese environment. Endosulfan usage in China has been estimated to be approximately 25700 t between 1994 and 2004. Concentrations of endosulfan in different environmental compartments in China, such as air, soil, water, and biota, but focusing at air and surface soil, have been summarized. Concentrations of total endosulfan in surface soil across China were ranged from below detection limit (BDL) to 19000 pg·g⁻¹ dry weight (dw), with geometric mean of 120 pg·g⁻¹dw. The results indicated that endosulfan sulfate had highest concentration in Chinese soil, followed by β- and α-endosulfan. Air concentrations of endosulfan in China were ranged 0–340 pg·m⁻³ for α-endosulfan and 0–121 pg·m⁻³ for β-endosulfan, with high concentrations occurred in the cotton production areas. Gridded usage inventories of endosulfan on a fine gridded system with a 1/4° longitude by 1/6° latitude resolution were compiled, from which, emission to air and residues in soil of endosulfan were calculated in each grid by using a modified simplified gridded pesticide emission and residue model (SGPERM), an integrated modeling system combining mathematical model, database management system, and geographic information system. Total emissions were around 10800 t from 1994 to 2004. Based on the emission and residue inventories, concentrations of α- and β-endosulfan in Chinese air and agricultural surface soil were also calculated for each grid cell, which are in general consistent with the published monitoring data.     

硫丹对蚯蚓存活、生长和肠胃线粒体超微结构的影响

通过人工土壤法对蚯蚓进行14d的急性暴露,研究了硫丹对蚯蚓存活、生长的影响;试验结束后,从硫丹剂量为0.5mg·kg-1和5 mg·kg-1的处理中挑选蚯蚓制作超薄切片,观察肠、胃部线粒体超微结构的变化.结果表明,硫丹对蚯蚓有较强的毒性作用,14d的LC50为6.52mg·kg-1.试验期间硫丹对蚯蚓的生长有显著的抑制作用,生长抑制率随硫丹剂量和暴露时间增加而增大.蚯蚓肠胃部的线粒体对硫丹有较强的敏感性,在非致死剂量下超微结构的损伤程度随硫丹剂量增加而加重.     

水质基准两栖类受试生物筛选

两栖动物是水生生态系统的重要生物群落,是水生生物基准的重要保护对象. 参照美国水生生物基准技术指南,搜集、筛选了12种本土代表性两栖动物的生物毒性数据,通过毒性数据分析,筛选出对两栖动物毒性最大的污染物,主要包括农药、重金属和杀虫剂三大类,以及4属基准研究受试生物. 在4属(5种)受试生物中,黑眶蟾蜍对硝酸银的累积概率为15%,六趾蛙对马拉硫磷的累积概率为1%,牛蛙对五氯酚的累积概率为8%,棘胸蛙对丙溴磷的累积概率为27%,虎纹蛙对硫丹的累积概率为29%. 结果表明:蛙属(六趾蛙)对农药、蛙属(牛蛙)对杀虫剂、蟾蜍属(黑框蟾蜍)对重金属均为敏感物种,虎纹蛙属(虎纹蛙)对农药、棘蛙属(棘胸蛙)对杀虫剂均为较敏感物种. 这4属(5种)两栖动物可以作为相关污染物的水质基准研究的受试物种.      

硫丹致生精细胞凋亡及其机制研究

即将列入持久性有机污染物(persistent organic pollutants,POPs)的硫丹对生物体的影响已经成为国际关注的焦点.为探讨硫丹对生精细胞凋亡影响及机制,以7.0、3.5和1.75 mg/kg剂量灌胃,连续染毒小鼠28 d.采用激光共聚焦扫描显微镜观察到染毒组小鼠凋亡的生精细胞形态学特征,二苯胺法测定到各染毒组DNA降解率显著升高(p＜0.01).利用Fluo-3能特异性地与细胞内钙离子结合并激发产生激光的特性,采用激光共聚焦扫描显微镜观察到染毒后小鼠生精细胞内游离钙离子浓度升高(p＜0.01).紫外分光光度法检测到生精细胞Na K -ATPase及Ca2 Mg2 -ATPase活力显著下降(p＜0.01),结果硫丹可诱导睾丸生精细胞凋亡.说明硫丹可抑制Na K -ATPase及Ca2 Mg2 -ATPase活力,使细胞膜主动转运Ca2 的功能受限,细胞的排钙能力降低,导致细胞内Ca2 浓度持续升高,即细胞钙超载,继发细胞钙稳态失衡.     

藻毒素对鱼类肝脏的毒理学效应

以鱼类肝脏为靶器官,研究了藻毒素对鱼类的毒性影响.结果表明,藻毒素在注射和口服两种不同的染毒方式下,均可引起受试鱼类发生急性中毒死亡.对中毒鱼类进行解剖分析发现,受试鱼类的肝/体重比高于对照组.说明无论哪种给药方式,藻毒素都会迅速地、专一地作用于鱼类肝脏,引起鱼类肝脏充血红肿,出现血斑.通过对鱼类肝脏组织碱性磷酸酶(ALP)同功酶谱的分析,可以发现在两种不同染毒条件下藻毒素各剂量组的ALP同功酶谱与对照组相同,均有ALP₁1～ALP₅5条带,只是高剂量组较低剂量组ALP₁～ALP₃ 3条带的着色深,并可使ALP同功酶各组分的相对活性上升.试验结果显示,藻毒素具有极高的细胞选择性和专一生物活性,其在生物机体内所引发的许多细胞学变化主要是对生物体肝脏酶系统的破坏所致,必须引起人们的重视.     

Small queens and big-headed workers in a monomorphic ponerine ant

Evolution of caste is a central issue in the biology of social insects. Comparative studies on their morphology so far suggest the following three patterns: (1) a positive correlation between queen–worker size dimorphism and the divergence in reproductive ability between castes, (2) a negative correlation among workers between morphological diversity and reproductive ability, and (3) a positive correlation between queen–worker body shape difference and the diversity in worker morphology. We conducted morphological comparisons between castes in Pachycondyla luteipes, workers of which are monomorphic and lack their reproductive ability. Although the size distribution broadly overlapped, mean head width, head length, and scape length were significantly different between queens and workers. Conversely, in eye length, petiole width, and Weber’s length, the size differences were reversed. The allometries (head length/head width, scape length/head width, and Weber’s length/head width) were also significantly different between queens and workers. Morphological examinations showed that the body shape was different between queens and workers, and the head part of workers was disproportionately larger than that of queens. This pattern of queen–worker dimorphism is novel in ants with monomorphic workers and a clear exception to the last pattern. This study suggests that it is possible that the loss of individual-level selection, the lack of reproductive ability, influences morphological modification in ants.     

微塑料对鲫鱼生长、肝脏损伤和肠道微生物组成的影响

广泛存在于自然环境中的微塑料可能会危害水生生物的生长和健康,目前该方面的研究较少.以淡水杂食性鱼类鲫鱼作为研究对象,喂食不同微塑料浓度的鱼食进行为期30 d的食源暴露试验,分析微塑料对鲫鱼生长、肝脏损伤和肠道微生物组成的影响.结果表明,与对照组相比,各试验组中鲫鱼的身长没有明显变化,但低浓度组的鲫鱼体重显著增加,中浓度组和高浓度组的体重明显下降.低浓度组的鲫鱼肝脏组织基本正常,中浓度组和高浓度组的均出现了不同程度的肝组织异常现象,其中高浓度组的肝脏损伤最为严重.鲫鱼肠道中,变形菌门、梭杆菌门、厚壁菌门和拟杆菌门是门水平上的优势菌种,葡萄球菌属和劳尔氏菌属等致病菌在各试验组鲫鱼肠道中均有出现;α多样性结果表明高浓度组的鲫鱼肠道微生物菌群最为丰富;PCoA结果显示对照组和试验组的鲫鱼肠道微生物存在明显的聚类特征.     

妊娠期全氟辛酸染毒对子代雄鼠发育毒性的研究

探讨妊娠期染毒全氟辛酸(perfluorooctanoic acid,PFOA)对子代雄鼠发育毒性的影响.将50只妊娠0 d小鼠随机分5组,每组10只,妊娠0~17 d每天按0、1、2.5、5和12.5 mg·kg~(-1)BW饲喂PFOA,产后仔鼠正常饲养,记录仔鼠存活数.产后21 d,检测仔鼠血清睾酮水平并计算睾丸指数;HE染色观察睾丸组织结构变化;RT-PCR法检测睾丸组织Dlk1-Dio3印记基因簇各目的基因mRNA的表达情况.结果显示,随PFOA染毒剂量增加,仔鼠存活数显著降低,血清睾酮含量极显著性降低(p0.01);睾丸指数无统计学意义;睾丸组织有不同程度的损伤并呈剂量依赖性;Dlk1-Dio3印记基因簇各目的基因的表达量都有降低趋势,其中Gtl2、Rian、Dio3均显著性降低(p0.05).结果表明,妊娠期染毒PFOA能够降低仔鼠存活数,损伤睾丸组织结构,并能够扰乱生殖激素,降低睾丸组织Dlk1-Dio3印记基因簇各目的基因mRNA的表达.     

扬州市长江段不同水层中鱼类有机氯农药残留研究

鱼肉含有的蛋白质,脂肪,矿物质,维生素对人的健康起到重要的作用,因而被大多数人所青睐。由于有机氯农药具有较高的杀虫活性,曾被广泛使用。中国早已停产,但由于其性质稳定在自然界不易分解属于高残留农药,残留于植物中有机氯农药,可通过＂食物链＂或其他途径进入动物体内,鱼类也会受影响。本文根据淡水鱼在水中分布状况,选取了扬州市长江段生活在不同水层中三种鱼类为分析对象。选取容易富集有机氯农药的腹部肌肉为实验样品测定部位,采用加速溶剂萃取法前处理分析鱼肉中有机氯农药并对该分析方法进行完善,通过实验总结出能满足环境监测工作中对淡水河流中鱼类有机氯农药测定,并能及时掌握淡水鱼类中有机氯农药对人身健康状况的影响。     

Mutagenic and cytotoxic potential of Endosulfan and Lambda-cyhalothrin - In vitro study describing individual and combined effects of pesticides

Excessive use of pesticides poses increased risks to non target species including humans. In the developing countries, lack of proper awareness about the toxic potential of pesticides makes the farmer more vulnerable to pesticide linked toxicities, which could lead to diverse pathological conditions. The toxic potential of a pesticide could be determined by their ability to induce genetic mutations and cytotoxicity. Hence, determination of genetic mutation and cytotoxicity of each pesticide is unavoidable to legislate health and safety appraisal about pesticides. The objective of current investigation was to determine the genotoxic and cytotoxic potential of Endosulfan（EN） and Lambda-cyhalothrin（LC）; individually and in combination. 3-（4,5-Dimethylthiazol-2-yl）-2,5-Diphenyltetrazolium Bromide（MTT） assay was utilized to determine cytotoxicity, while two mutant histidine dependent Salmonella strains（TA98, TA100） were used to determine the mutagenicity of EN and LC.Moreover, mutagenicity assay was conducted with and without S9 to evaluate the effects of metabolic activation on mutagenicity. Even though a dose dependent increase in the number of revertant colonies was detected with EN against both bacterial strains, a highly significant（p 〈 0.05） increase in the mutagenicity was detected in TA98 with S9. In comparison, data obtained from LC revealed less mutagenic potential than EN. Surprisingly,the non-mutagenic individual-concentrations of EN and LC showed dose dependent mutagenicity when combined. Combination of EN and LC synergistically induced mutagenicity both in TA98 and TA100. MTT assay spotlighted comparable dose dependent cytotoxicity effects of both pesticides. Interestingly, the combination of EN and LC produced increased reversion and cytotoxicity at lower doses as compared to each pesticide, concluding that pesticide exposure even at sub-lethal doses can produce cytotoxicity and genetic mutations, which could lead to carcinogenicity.     

Morphometric relationships and reproductive maturation of the mudskipper,Periophthalmus barbarus from subsistence catches in the mangrove swamps of IMO estuary,Nigeria

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沿江化工园区污水处理厂出水对体外培养的大鼠睾丸细胞生殖毒性研究

应用基于原代培养的大鼠睾丸细胞体外毒性测试方法,通过支持细胞、生精细胞及间质细胞这3种细胞的形态、活力、胞外乳酸脱氢酶（LDH）活力、细胞早期凋亡坏死率和睾酮分泌量等指标对沿江化工园区污水处理厂出水的生殖毒性进行了研究. 结果表明,新开发的沿江化工园区A和省级沿江化工园区B污水处理厂出水有机提取物浓缩后均能造成不同程度的大鼠睾丸细胞生殖毒性,园区A提取物能明显影响支持细胞（p<0.01）、生精细胞（p<0.05）和间质细胞（p<0.05）的细胞活力,影响支持细胞和生精细胞胞外LDH活力（p<0.01）,且其毒性明显高于园区B. 暴露导致支持细胞处于凋亡早期,而生精细胞比较敏感,已到了凋亡晚期甚至有小团细胞坏死,虽然间质细胞的形态没有显著变化但园区A有机提取物已明显抑制了间质细胞睾酮的分泌（p<0.01）. 研究表明,通过基于原代培养的大鼠睾丸细胞体外毒性测试方法能够对污水处理厂出水的生殖毒性做出综合评价,该方法可以作为现行国家污水综合排放化学监测的有效补充.     

Cytochrome P450 BM3 of Bacillus megaterium—A possible endosulfan biotransforming gene

Computing chemistry was applied to understand biotransforrnation mechanism of an organochlorine pesticide, endosulfan. The stereo specific metabolic activity of human CYP-2B6 （cytochrome P450） on endosulfan has been well demonstrated. Sequence and structural similarity search revealed that the bacterium Bacillus megaterium encodes CYP-BM3, which is similar to CYP-2B6. The functional similarity was studied at organism level by batch-scale studies and it was proved that B. megaterium could metabolize endosulfan to endosulfan sulfate, as CYP-2B6 does in human system. The gene expression analyses also confirmed the possible role of CYP-BM3 in endosulfan metabolism. Thus, our results show that the protein structure based in-silico approach can help us to understand and identify microbes for remediation strategy development. To the best of our knowledge this is the first report which has extrapolated the bacterial gene for endosulfan biotransformation through in silico prediction approach for metabolic gene identification.     

Methyl farnesoate stimulates testicular growth in the freshwater crab Oziotelphusa senex senex fabricius

 The influence of methyl farnesoate (MF) on testicular growth in the fresh water crab Oziotelplusa senex senex was studied. MF stimulated testicular growth as evidenced by increased testicular weight, testicular index and testicular follicle diameter in MF injected crabs and provides evidence that MF acts as a male reproductive hormone in crustacea. Received: 25 August 1998 / Accepted: 18 February 1999     

溴氰菊酯混配农药--大地西对稻田-鱼塘水生生态系的安全评价研究

研究了在自然条件下施用溴氰菊酯混配农药——大地西对稻田-鱼塘水生生态系的影响。结果表明,在稻田中施用时,由于实际用量低,加上作物的沾附、土壤与水体中悬浮物的吸附及其存在形态的变化,其实际浓度与毒性已降低到对稻田养鱼及邻近鱼塘的水生生物不再有危害的程度。描述了溴氰菊酯在整个模拟生态系中的迁移与消减规律。研究了溴氰菊酯在不同加工剂型中的存在形态,以及在鱼体内的吸收与分布特点及其与毒性的关系。     

Behavioural and physiological consequences of male reproductive trade-offs in edible dormice (Glis glis)

Testosterone mediates male reproductive trade-offs in vertebrates including mammals. In male edible dormice (Glis glis), reproductivity linked to high levels of testosterone reduces their ability to express torpor, which may be expected to dramatically increase thermoregulatory costs. Aims of this study were therefore to analyse behavioural and physiological consequences of reproductive activity in male edible dormice under ecologically and evolutionary relevant conditions in the field. As we frequently encountered sleeping groups in the field, we hypothesized that social thermoregulation should be an important measure to reduce energy expenditure especially in sexually active male edible dormice. Our results revealed that the occurrence of sleeping groups was negatively influenced by male body mass but not by reproductive status or ambient temperature. In reproductive as in non-reproductive males, the number of individuals huddling together was negatively influenced by their body mass. Thus in general males with a high body mass were sitting in smaller groups than males with a low body mass. However, in reproductive males group size was further negatively affected by ambient temperature and positively by testes size. Thus breeders formed larger sleeping groups at lower ambient temperatures and males with larger testes were found in larger groups than males with smaller testes. Measurements of oxygen consumption demonstrated that grouping behaviour represents an efficient strategy to reduce energy expenditure in edible dormice as it reduced energy requirements by almost 40%. In summary, results of this field study showcase how sexually active male edible dormice may, through behavioural adjustment, counterbalance high thermoregulatory costs associated with reproductive activity.