Characterization of cholinesterases in marbled sole, Limanda yokohamae, and their inhibition in vitro by the fungicide iprobenfos

Cholinesterases (ChEs) have been characterized in marbled sole (Limanda yokohamae) for use as a possible biomarker of pollution exposure. In brain, ChEs existed almost exclusively (>95%) as acetylcholinesterase (AChE) whereas in muscle, about 20-30% of ChE activity was in the form of butyrylcholinesterase (BChE; pseudocholinesterase). Acetylthiocholine and butyrylthiocholine (identified in mammalian studies as diagnostic substrates for AChE and BChE respectively) were hydrolyzed mainly, but not exclusively, by these enzymes. The inhibitors BW284C51 and iso-OMPA (identified in mammalian studies as diagnostic inhibitors of AChE and BChE respectively) were not specific for these enzymes in marbled sole. Brain AChE and muscle AChE and BChE were characterized in terms of their kinetic properties (KM etc.) and optimal conditions (substrate concentration, protein concentration, pH etc.) were established to allow routine assays of ChE activity to proceed under pseudo-first order conditions. The sensitivity of ChEs to a locally significant pesticide, iprobenfos (IBP; kitazin) was established in terms of IC50 concentrations. Brain AChE was relatively insensitive to IBP, but muscle AChE and BChE were sensitive to IBP concentrations in the high nM range. However, ambient IBP concentrations in Korean coastal waters are usually not high enough to cause detectable ChE inhibition in this species.     

海鱼AChE监测海水有机磷农药的可行性研究

有机磷农药(OPs)对乙酰胆碱酯酶(AChE EC 3.1.1.7)具有专一性抑制作用,因此AChE常作为淡水和土壤OPs污染的特异性标识物(biomarker).本文在海水背景下,通过测定两种有机磷农药(马拉硫磷和甲基对硫磷)对海洋鱼类-鲈鱼[Lateolabrax japonicus(cuvier)]脑组织AChE的抑制效应,分析了海鱼AChE监测海水OPs的可行性.结果显示,抑制时间为20min时,抑制程度达到相对稳定;在低浓度OPs抑制下,纯酶液比粗酶液表现出更好的敏感性,对马拉硫磷和甲基对硫磷的灵敏度分别达到0.28μg*L-1和0.2μg*L-1;在一定浓度范围内,AChE活力对OPs具有显著的线性响应.研究表明,利用鲈鱼AChE指示和监测海水OPs的微污染是完全可行的.     

三价砷和五价砷对鱼类乙酰胆碱脂酶的体外抑制比较

为开展水产品中三价砷的酶法快速检测工作,文中研究了三价砷(As3 + )、五价砷(As5+ )对8种鱼脑中乙酰胆碱酯酶(AChE)的抑制率。结果发现,在10 -7～10 -4mol/L范围内,草鱼和鲫鱼脑中AChE的抑制率与三价砷的浓度负对数值有较好的线性关系,相关系数都在0 .97以上;在10 -6～10 -4mol/L范围内,黑、马鲛、鲈鱼脑中AChE的抑制率与三价砷的浓度负对数值的线性相关系数大于0 .99。对于三价砷,由IC50 得到AChE的敏感性为:美国红鱼>草鱼,鲫,鲈鱼>黑,马鲛>鲤,白鲢。鱼种不同,AChE受到的三价砷、五价砷的抑制作用也不同。在浓度为10 -6～10 -4mol/L范围内,三价砷、五价砷对8种鱼脑中AChE的抑制作用存在极显著差异(p <0 .0 1) ,三价砷对AChE的抑制作用远大于五价砷的抑制作用。     

Role of biotransformation in determining aldicarb toxicity in fish

The carbamate pesticide, aldicarb, demonstrates significant acute toxicity in fish, and is readily biotransformed by most organisms studied. In fish, both the cytochrome P450 (CYP) and the flavin monooxygenase systems (FMO) are involved in bioactivating aldicarb to aldicarb sulfoxide, which is a more potent inhibitor of acetylcholinesterase (AChE). Channel catfish (Ictalurus punctatus), along with many other fresh water species, do not express FMO and are relatively resistant to the effects of aldicarb. This project examined the toxicity, AChE inhibition, metabolism, and toxicokinetic of aldicarb in channel catfish, and compared these values with an aldicarb-sensitive species, rainbow trout, which expresses FMO. Studies of in vitro and in vivo aldicarb biotransformation in catfish suggest that a low rate of bioactivation (10 times slower V_max), resulting in less initial conversion to the activated metabolite, aldicarb sulfoxide, may be a contributing factor to resistance of channel catfish to aldicarb toxicity. These data are supported by toxicokinetic and enzyme inhibition studies. This work demonstrates that differences in FMO expression among fish species may have significant influence on toxicity resulting from exposure to some xenobiotics.     

Kinetic and toxicological characteristics of acetylcholinesterase from the gills of oysters (Crassostrea rhizophorae) and other aquatic species

The aim of this work was to characterize the cholinesterases from gills of Crassostrea rhizophorae in order to use them as biomarkers. Gills were homogenized and then centrifuged (9,000 x g, 4 degrees C, 30 min). S9 and Triton X-100 S9 treated (TX S9) fractions were employed as enzyme source. Km(ap) and Vmax were estimated, using acetylthiocholine iodide as substrate. Inhibition assays were performed with iso-OMPA and eserine. The Km(ap) for S9 and TX S9 fractions were 0.05 and 0.06 mM, whereas the Vmax were 1.92 and 5.84 nmol/min/mg protein. respectively. No inhibition was detected when the samples were incubated with iso-OMPA, suggesting the presence of acetylcholinesterases (AChE) in oyster gill homogenates. Sensitivity to eserine inhibition of AChE in the gills of oysters is intermediate when compared with other aquatic species.     

几种鱼类培养细胞对不同干扰素敏感性的研究

本文研究了四种鱼类培养细胞ＣＩＫ、ＣＡＢ、ＧＣＢ、ＢＲＥ对Ｈｕｉｆｎ－ａ,ＣＩＫ、ＣＡＢ、ＧＣＢ、ＧＲＥ、ＦＨＭ等五种鱼类培养细胞干扰素作用的敏感性。这四种鱼类培养细胞对ＨｕＩＦＮ－ａ不敏感,对五种鱼类培养细胞干扰素表现出相对一致的敏感性差异,其中ＧＲＥ细胞对鱼类干扰素最敏感,其次为ＣＡＢ、ＧＣＢ。ＣＩＫ细胞能产生高滴度的干扰素但对鱼类干扰素不敏感。不同病毒对干扰素作用的敏感性也不同。     

Effects of naphthalene and phenanthrene on visual and prey capture endpoints during early stages of the dourado Salminus brasiliensis

Naphthalene (NAP) and phenanthrene (PHE) effects on Salminus brasiliensis, a carnivorous freshwater fish, were investigated using behavioral tests. Larval stages of S. brasiliensis were exposed to water concentrations of 0, 0.04 mg/l, 0.20mg/l and 0.50mg/l for naphthalene and 0, 0.01 mg/l, 0.05 mg/l and 0.1mg/l for phenanthrene during two developmental phases. The prey fish Prochilodus lineatus were not exposed. Visual acuity was measured at the end of phase 2 in individual S. brasiliensis, which were also tested at the end of each phase for number of attacks on prey, number of prey captured, prey capture efficiency, and distance swam. Vision was impaired by PHE exposure, as acuity angles increased in exposed fish. At Stage I 2.3+/-1.2 prey were captured with 46% efficiency in controls compared to 0.4+/-0.3 prey captured with 13.4% efficiency in fish exposed to 0.05 mg PHEl(-1), the LOEL for both endpoints. At Stage II 4.0+/-1.1 preys were captured in controls compared to 2.5+/-0.9 preys captured in fish exposed to 0.01 mg PHEl(-1), the LOEL. Stage II control fish captured prey with 70% efficiency compared to 30% efficiency at Stage II fish exposed to 0.05 mg PHEl(-1), the LOEL. Distance swam was not affected by either NAP or PHE exposure. The exposure of larval stages of S. brasiliensis to realistic water concentrations of PHE impairs foraging skills and could affect recruitment of the species.     

Assessing neuroprotective P-glycoprotein activity at the blood-brain barrier in killifish (Fundulus heteroclitus) using behavioural profiles

The neuroprotective role of P-glycoprotein, a multixenobiotic resistance transporter (ABCB1/MDR1), in the blood-brain barrier in fish was examined using behavioural toxicological assays. P-glycoprotein acts as cellular efflux pump to prevent substrates from accumulating in the brain, including environmental contaminants such as ivermectin, a common aquaculture pesticide and mammalian anti-parasitic drug. The behavioural toxicological assays were developed to determine the neuropathological effect of ivermectin in killifish (Fundulus heteroclitus). P-glycoprotein function and thus blood-brain barrier integrity can be compromised by chemosensitizers that inhibit transport activity. Fish treated with ivermectin and the P-glycoprotein inhibitor cyclosporin A were significantly more sensitive and succumbed more rapidly to tilting, lethargy, slowing of pectoral-fin movement and loss of haptic-reactivity compared to fish treated with ivermectin-only. P-glycoprotein inhibition is associated with significantly earlier onset and increased mortality in ivermectin-exposed fish. Our results suggest that P-glycoprotein confers resistance against ivermectin-induced behavioural neuropathology and mortality in fish. This assay provides us with a non-invasive tool to study P-glycoprotein function in the blood-brain barrier and evaluate the behavioural effects of potential environmental neurotoxins.     

Species-dependent sensitivity to contaminants: an approach using primary hepatocyte cultures with three marine fish species

There is limited knowledge about the sensitivity of different fish species to environmental pollutants. Such information is pivotal in risk assessment and to understand why some species appear to be more tolerant to contaminants than others. The aim of the current study was to evaluate whether primary hepatocyte cultures of three marine fish species could be established in the field and whether their sensitivity to selected contaminants would differ. Primary hepatocyte cultures of three marine fish species (plaice, long rough dab, Atlantic cod) were established and exposed for 24 h to copper (20-2500 mg L?1) and statins (1-200 mg L?1). Endpoints were esterase activity, metabolic activity and reduced glutathione (GSH) content, all using fluorescent probes. Flatfish hepatocytes were more susceptible to copper and statin exposure than hepatocytes from cod. This study has shown that species-dependent differences in contaminant sensitivity can be investigated using primary hepatocyte cultures.     

Inhibition of coral fertilisation and larval metamorphosis by tributyltin and copper

Fertilisation and larval metamorphosis of reef-building corals are important life history events leading to recruitment of juvenile corals to reef populations. Little is known of the sensitivity of these early life phases to pollution, or their relative susceptibility to certain toxicants compared with established coral colonies. Inhibition of fertilisation and larval metamorphosis of the coral Acropora millepora (Ehrenberg, 1834) was assessed in response to solutions of the antifoulants tributyltin (TBT) and copper (Cu) using laboratory-based bioassays. Nominal concentrations that inhibited 50% fertilisation and metamorphosis (IC50) were calculated from 4 h fertilisation and 24 h metamorphosis assays and were based on introduced dose. Cu was most potent towards fertilisation with an IC50 of 17.4 micrograms/l. TBT however, proved more toxic to larval metamorphosis having an IC50 of 2.0 micrograms/l. Inert surfaces coated with either Cu- or TBT-based antifouling paint also inhibited fertilisation and metamorphosis. The degree of inhibition was correlated with surface area of the paint coating. These results indicate fertilisation and metamorphosis of coral can be sensitive to active components of antifouling paints.     

中国多金属结核开辟区仔稚鱼的分布特征

利用 1 997年～ 1 999年“大洋一号”调查船在中国多金属结核开辟区用邦哥网所采集的样品 ,对仔稚鱼的种类组成、密度的平面分布和年间变化及其与环境因子的关系进行了研究。结果表明 :中国开辟区共有 2 2科 6 1种仔稚鱼 ,其中大洋性发光鱼类灯笼鱼科的种类最多 ,达1 7种 ,约占总种类数的 2 8% ,其次是钻光鱼科 (6种 )、金枪鱼科 (6种 )和拟白鲑科 (5种 ) ,分别约占总种类数的 1 0 %、 1 0 %和 8%。东、西两区的种类组成有较大的差异 ,共有种仅为 1 7种 ,东区种类数 (48种 )明显多于西区种类数 (3 0种 )。仔稚鱼的密度有明显的空间和时间变化 ,东区仔稚鱼的密度高于西区 ,东区 1 999年仔稚鱼的密度约为 1 997年的 5倍 ,西区 1 999年仔稚鱼的密度约为 1 998年的 3倍。仔稚鱼密度的年间变化可能与 El Nino事件有关     

Biomarker responses to pollution in two invertebrate species: Scrobicularia plana and Nereis diversicolor from the Cádiz bay (SW Spain)

The clam Scrobicularia plana and the polychaete worm Nereis diversicolor were collected in several sites from a littoral enclosure in SW Spain. The aim of our study was to relate various biomarker responses in these species to a pollution gradient caused by untreated domestic discharges and to verify the adequacy of the selected species as sentinels in this habitat. The biomarkers selected were the antioxidant enzymes catalase (CAT), glutathione peroxidase (GPX) and DT-diaphorase (DT-D). In addition, the activities of cytochrome P450-dependent ethoxyresorufin O-deethylase (EROD) activity, the phase II detoxifying enzyme glutathione S-transferase (GST) and the neurotoxicity marker acetylcholinesterase (AChE) were measured. Metallothionein levels were selected as biomarkers of heavy metals exposure in both species. The results suggest a different response in the water filtering organism (clam) and the sediment eater (polychaete), probably as a consequent of different pollution exposure and that samples from the "Ca?o Sancti-Petri" were exposed to biologically active compounds that altered some of their biochemical responses. AChE was the most sensitive biomarker in both species and N. diversicolor proved to be a more robust sentinel in this ecosystem.     

Fish species diversity in the Huanghe Estuary

I~IOXThe Huanghe River is among the largest rivers in the world. It is the cradle of the ChineseciviliZation, and haS a great importance in fisheries. There were some studies on the HuangheRiver fisheries, such as the Repert of the Basic Fishery Biological Su~ of the Huanghai Riber(Anon, 1960), The Fishery Resources Of the Huanghe Riber System (Anon, 1986a). With regard to the fish species diversity in the Huanghe Estuary, there were only some general information from Read of Ichth…     

Environmental monitoring using acetylcholinesterase inhibition in vitro. A case study in two Mexican lagoons.

Cholinesterase inhibition is considered a specific biomarker of exposure and effect for organophosphorous pesticides. Its use for monitoring has been hindered, particularly in tropical countries where organophosphates are widely used for malaria and dengue control, because of the frequent lack of suitable controls. An in vitro technique is proposed as a biochemical method for monitoring pollutant mixtures in sediment toxicity tests. Brain homogenate from the fish Oreochromis niloticus is used as the enzyme source. Optimum incubation time, extraction solvent and effect of crude oil on acetylcholinesterase (AChE) are reported. The method described was used in sediments from two Mexican lagoons, located in an oil extraction area where pesticides are used in agriculture and vector control campaigns. AChE inhibitions from 3 to 21% were found in these lagoons, even in the presence of high concentrations of petroleum.     

黄河口及附近海域鱼卵和仔鱼种类组成及分布特征

根据2009年5月和8月2个航次对黄河口及附近海域18个站位的鱼卵和仔鱼调查资料,对该水域鱼卵仔鱼的种类组成和数量分布特征进行了分析和探讨.结果表明,鱼卵仔鱼的种类组成和数量分布呈现明显的季节变化特征;春季鱼卵仔鱼种类数高于夏季,平面分布范围大于夏季,主要优势种季节更替明显;与历史同期资料相比,鱼卵仔鱼种类和数量大幅下...     

厦门东海域定置网渔获鱼类种类组成及其季节变化

2003年7月～2004年6月对厦门东部海域的定置网渔获鱼类进行采样分析．结果表明厦门东海域至少有鱼类242种，隶属于19个目81科145属，主要是沿岸近底层和底栖的小型鱼类．以鲈形目种类最多，共计127种，占总数的52．5％、经济种类有116种．主要优势种有：青鳞小沙丁鱼、孔鳞小沙丁鱼、斑鲦、马拉邦虫鳗、裸鳍虫鳗、中华须鳗、前鳞鲻、眶棘双边鱼、多鳞鳝、皮氏叫姑鱼、短棘银鲈、列牙垒幸4、黄斑蓝子鱼、褐蓝子鱼、锯塘鳢、锻虎鱼类、褐菖鲇、绿鳍鱼、鳄鲡、卵鳎、中华单角鲍等．渔获种类在秋季及冬初最多，夏季较多，而冬末和春初最少．渔获种类的季节变化比较明显，与海洋表层水温变化有关，种类的月更替率比较高．     

Effects of BPA and DES on longchin goby (Chasmichthys dolichognathus) in vitro during the oocyte maturation

This study was performed in order to assess whether bisphenol (BPA) and diethylstilbestrol (DES) had agonistic or antagonistic effects on oocyte maturation using marine fish. We tested the effects of these chemicals on in vitro maturation, germinal vesicle breakdown (GVBD), assay using oocytes from the longchin goby, Chasmichthys dolichognathus. During the maturation process, low concentrations of BPA and DES triggered GVBD depending on the stage of oocyte development; BPA at 0.044 nM and DES at 0.037, 0.37, and 3.73 nM induced GVBD in 0.82-0.88 mm diameter oocytes (germinal vesicle located near the center of oocytes). In 0.76-0.80 mm diameter oocytes (fully vitellogenic oocytes), BPA induced GVBD at relatively higher concentrations (4.38, 43.8, and 438 nM). In 0.86-0.90 mm diameter oocytes, BPA and DES had no observable effect on GVBD at the concentrations tested. Oocytes with diameters between 0.82 and 0.88 mm appeared to be more sensitive to these chemicals. Moreover, our results showed that BPA and DES did not inhibit GVBD.     

长江口基础生物资源现状及年际变化趋势分析

探讨长江口生物资源可持续利用和对生物栖息环境的保护措施,2004年对长江口海域的基础生物资源进行4次监测。结果表明:浮游植物5月份有5门39属86种,8月份有6门48属139种,河口锋附近水域的浮游植物种类、数量较多,中肋骨条藻(Skeletonema costatum)在数量上处于绝对优势;浮游动物5月份为77种,8月份为91种,中小型浮游动物的数量在河口锋附近水域较高,主要优势种有针刺拟哲水蚤(Paracalanus aculeatus)等,大型浮游动物种类数量较高的区域位于长江冲淡水以外的海域,主要优势种为中华哲水蚤(Calanus sinicus)、五角水母(Muggiaea atlantica)、真刺唇角水蚤(Labidocera euchaeta);2004年共鉴定底栖生物146种,5月份63种,8月份108种,其中多毛类41种、甲壳类41种、软体动物32种、棘皮动物6种、鱼类21种、其他4种;长江口沿岸的崇明东滩、南汇边滩和嵊山岛共鉴定潮间带生物126种,其中5月份86种,8月份71种;鱼卵仔鱼计8目20科30属34种,鱼卵中绝大部分为凤鲚,其数量占95.7%,仔鱼以中华小公鱼出现频次最多。通过年际变化分析表明,长江口门以内区域浮游生物种类明显下降,长江冲淡水区域的生物资源基础饵料生物种类数基本保持稳定,浮游生物群落结构发生变化,硅藻比重减少,甲藻增加,长江口区域的潮下带和潮间带底栖生物种类数也存在不同程度的下降,以口门以内区域下降明显。     

高公岛近岸海域浮游植物群落特征

2009年8月对高公岛附近海域的海洋浮游植物及环境因子进行了调查,在14个调查站位共鉴定浮游植物4门36属81种,其中,硅藻门25属63种,占种数的77.78%;甲藻门9属16种,占种数的19.75%,蓝藻门1属1种,占种数的1.23%,金藻门1属1种,占种数的1.23%。浮游植物生态类型主要以低盐近岸性类群为主,辅以少量的广温广盐性类群。主要优势种为斯氏几内亚藻、短角弯角藻、拟旋链角刺藻、扭链角刺藻。优势度分别为0.688、0.207、0.142、0.103。浮游植物的细胞丰度范围为0.458×105～3.134×105个/L,平均值为1.180×105个/L。群落特征指数变幅较大(种类数为19～43,丰富度指数为1.064～2.364,多样性指数为2.069～3.428,均匀度指数为0.485～0.677)。丰富度指数均值为1.753多样性指数均值为2.926,均匀度指数均值为0.592。相关性分析表明,仅均匀度指数与水温之间具有显著的相关性,相关系数(r)为0.544,P<0.05,水温越高,均匀度指数越高;其它因子之间均没有显著的相关性。高公岛近岸海域水质状况的生物多样性指数评价显示,该海域水质总体处于轻污染,尚适宜浮游植物的生长。     

Effects of diel period and tow duration on estuarine fauna sampled with a beam trawl over bare sediment: Consequences for designing more reliable and efficient surveys

The effects of diel period and tow duration (5, 10 and 20 min) on samples of estuarine fauna in a beam trawl, were tested over bare sediment in Tuggerah Lake (New South Wales, Australia). Mean catch rates (numbers of fish caught 5 min⁻¹) were significantly larger at night for the total numbers of individuals and abundant, economically important species of fish and invertebrates (e.g. Gerres subfasciatus, Metapenaeus macleayi, Penaeus plebejus). Greater proportions of larger fish were also caught at night for some species (e.g. G. subfasciatus, Acanthopagrus australis, Rhabdosargus sarba), but not across all tow durations. Multivariate analyses detected dissimilarities in the composition and structure of assemblages between diel periods, which were driven by species caught predominately, or in larger proportions, at night. Short tows (5 min) were more efficient than longer tows (10 or 20 min) for sampling the diversity of species (i.e. most species were caught in the first 5 min of a tow). There were, however, no clear or consistent patterns relating to the effect of tow duration on the catch rates of other variables, the size ranges of abundant species, or the structure and composition of assemblages. Our data confirm that at night, bare sediment is an important habitat for a wide size- and species-range of estuarine fish and invertebrates. In future, more cost-effective and reliable information concerning these taxa would be achieved by sampling with the beam trawl at night, using tow durations of 5 min. We also highlight a problem inherent in the design of many studies of diel variation of fauna (i.e. the potential non-independence of data among day and night periods) and discuss its solution.