Residues of [14C]-malachite green in eggs and fry of rainbow trout, Oncorhynchus mykiss (Walbaum), after treatment of eggs

Abstract. Rainbow trout, Oncorhynchus mykiss (Walbaum), eggs were exposed to [methane-¹⁴C] malachite green chloride on day 0 and on every third day thereafter through day 24, with a final treatment administered to fry on day 31. Eggs or fry were sampled before each treatment, and at selected times from day 31 to day 59. Malachite green equivalence in eggs and fry was determined by sample oxidation and liquid scintillation counting. Total malachite green residues increased throughout the exposure period to 0.271 ± 0.042 μg g⁻¹ (± SD) on day 31. Residues declined to 0.055 ± 0.011 μg g⁻¹ on day 59. The depuration phase declined monoexponentially with a half-life of 13.3 days for the absolute amount (μg sample⁻¹) and a half-life of 9.7 days for the concentration of malachite green residues (μg g⁻¹). Growth dilution accounted for the 25% increase in the elimination (9.7 days) of malachite green residues. Extracts from treated eggs and fry were analysed by reversed-phase liquid chromatography. Three peaks were resolved in treated eggs: chromatic malachite green, leuco malachite green and an unknown polar metabolite. Only two peaks were resolved in the fry: leuco malachite green and the unknown polar metabolite. The most prominent residue in all samples was leuco malachite green.     

水产品中孔雀石绿检测方法研究进展

水产品的安全问题日益成为社会关注的焦点,近年来,我国出口的鳗鲡以及国内市场上的水产品都发现了孔雀石绿残留.国内各有关检测机构相继开展了水产品中孔雀石绿残留量的检测.本文对现有的各种检测方法的优点和局限性作一概述,以便研究者能在此基础上进一步改进和发展.     

The Toxicity of Erythromycin, Minocycline, Malachite Green, and Formalin to Nauplii of the Shrimp Penaeus stylirostris

The toxicities of erythromycin, minocycline, malachite green, and formalin to nauplii of the shrimp Penaeus stylirostris were determined in a static bioassay. Toxicity was assessed on the basis of survival of nauplii after 12 and 24 hours of exposure to the compounds and metamorphosis of the nauplii to protozwa. The results suggested that metamorphosis to protozoea is more susceptible to toxic effects than is naupliar survival. Metamorphosis was either reduced or completely inhibited by lower concentrations of erythromycin, minocycline, and malachite green than was naupliar survival at 12 or 24 hours. Metamorphosis was either reduced or completely inhibited by erythromycin, minocycline, malachite green, and formalin concentrations of 80, 100, 0.08, and 27 mg/ L, respectively. Toxic effects were not observed at erythromycin, minocycline, malachite green, and formalin concentrations of 16, 62.5, 0.016, and 2.7 mg/L, respectively. The results suggest that formalin may be toxic at therapeutic levels frequently recommended for post larvae and older penaeids, but that erythromycin, minocycline and malachite green are not.     

Acclimation of the brown trout, Salmo trutta L., to the stress of daily exposure to malachite green

Exposure of brown trout to a single dose of malachite green (2.27 ppm) caused a significant elevation of blood cortisol levels within 1 h. After 4 weeks of daily exposure to malachite green, however, this stress response was totally abolished. Daily treatment had no long-term effects on basal plasma cortisol levels, or on the numbers of circulating erythrocytes, thrombocytes and lymphocytes. Blood neutrophil numbers were significantly reduced in malachite green-treated fish, an effect thought to be due to a decreased ectoparasitic load.     

不同冷冻贮藏时间对三种鱼类肌肉中孔雀石绿残留量的影响

研究鲤鱼、鳗鲡、鳜鱼在加入不同浓度的孔雀石绿标准样品且在不同冷冻贮藏时间下对孔雀石绿残留量的影响。三种基质中,鳗鲡组织中添加有无色孔雀石绿降解速率最慢、鳜鱼次之、鲤鱼较快。三种基质中添加10μg/kg、50μg/kg、100μg/kg有色孔雀石绿平均降解率分别为57.9%、48.0%、23.0%;无色孔雀石绿平均降解率分别为46.6%、43.4%、19.8%,有色孔雀石绿较无色孔雀石绿降解快,低浓度较高浓度降解快。在三种基质鲤鱼、鳗鲡、鳜鱼组织中有色和无色孔雀石绿降解反应均符合准一级反应动力学。分析可能由于鳗鲡组织脂肪较多,其结合方式影响了有色无色孔雀石绿的降解。     

孔雀石绿的污染治理及样品中残留物的分离检测方法

孔雀石绿是一种三苯甲烷类化合物,作为一种重要的化工原料、碱性染料、杀真菌剂、生物染色剂得到了广泛的应用.由于其毒性和可致癌性,已经被禁止用于水产品生产和运输过程中.综述了孔雀石绿污染物的治理和样品中其残留物的分离检测方法.     

An in vitro method for testing synergistic action of parasiticides using malachite green and formalin as a model system

Abstract. An in vitro technique was developed to screen drugs for toxicity to laboratory cultured protozoans. Drugs were diluted using standard microtitration equipment. An easily cultured protozoan, Tetrahymena pyriformis , was added to the dilutions of drugs and viability was checked daily for 10 days. Drugs used as a model system to test the technique were formalin and malachite green. Effective concentrations of formalin determined by this in vitro technique were close to recommended therapeutic treatment levels for Ichthyophthirius . Malachite green was effective at concentrations in excess of recommended therapeutic treatment levels for Ichthyophthirius . The synergistic effect of formalin and malachite green was demonstrated. This technique can he of use in determining effectiveness of new drugs as parasiticides.     

Malachite green: a review

Abstract. A detailed review of malachite green and its chemical and physical properties as they relate to fisheries interests. Many of the confusions and misconceptions about malachite green are discussed. Its mode of action, toxicity and fate are considered.     

渔药孔雀石绿及其关联化合物检测方法研究进展

主要对孔雀石绿、结晶紫及其代谢物之间的关系、毒性和检测方法现状作一概述。对各种检测方法的差异进行了比较，以便研究者能在此基础上进一步发展改进现有的检测方法。     

Sensitivity of juvenile lobsters, Homarus americanus L., to different concentrations of malachite green during various states of the intermoult cycle

Abstract. Survival of the early stages (egg, larva and juvenile) of the American lobster, Homarus americanus L., reared under controlled conditions has often been threatened by a variety of fungi and microbial epibionts. A preliminary study was conducted for the purpose of defining toxicity levels of malachite green for 5th stage juveniles during each of the following intermoult (or fused intermoult) states: A-B, C₄, D₀ and D₂-D₃ Irrespective of exposure time or concentration, all the lobsters treated during intermoult state A-B died. This contrasts with seemingly good survival of lobsters in intermoult states C₄, D₀ and D₂- D₃ treated at concentrations of 900 mg/1 malachite green for exposure periods of 10 min. The findings suggest that an effective treatment combination for lobsters immersed in malachite green may be one which is employed during intermoult state C₄, for about 6min and at concentrations of about 700–800 mg/l.     

Toxicity of malachite green to cultured american lobster larvae

Malachite green has been used to control fungal and epibiotic infestations of larval lobsters. The concentration of malachite green and the exposure period are critical to survival. The effects of increasing concentrations and exposure periods show survival to be adversely effected with treatments above 8 ppm for 16 min and above 20 ppm for 8 min administered every other day throughout the larval period. Development of the larvae is retarded with increasing treatment levels. Survivors of high treatment levels show no immediate post-treatment effects.     

Malachite green: a pharmacokinetic study in rainbow trout, Oncorhynchus mykiss (Walbaum)

Abstract. Pharmacokinetic studies on uptake, distribution and climination of malachite green in rainbow trout, Oncorhynchus mykiss (Walbaum), are described. All three processes were found to be temperature-dependent, with levels and rates increasing with increases in water temperature. Tissues investigated were serum, liver, kidney, muscle and viscera. First-order kinetics were found to apply and, in particular, kidney was observed to be a deep compartment. The long-term persistence of kidney residues may explain the efficacy of malachite green for the control of proliferative kidney disease in rainbow trout. Residue levels in muscle were never excessive and an extrapolation from the experimental data suggests that a suitable withholding period might be 600 degree days.     

Evaluation of the secondary stress response in healthy Nile tilapia (Oreochromis niloticus L.) after treatment with a mixture of formalin, malachite green and methylene blue

To determine the effects of exposure to a mixture of formalin, malachite green and methylene blue (FMC) on the secondary stress indices, changes in glucose, phosphorus, calcium, magnesium, sodium, potassium and haematocrit were monitored in healthy Nile tilapia, Oreochromis niloticus L. Fish were exposed separately to varying concentrations of a mixture of formalin, malachite green and methylene blue (0.1, 0.5 and 1 p.p.m.) for 1, 10 and 60 min. In general, treatment of fish with FMC elicited marked elevations of plasma glucose. Plasma phosphorus levels dropped after FMC treatment. In fish exposed to FMC, calcium levels in general were lower than those of the controls. Magnesium levels were not influenced by FMC treatment. Plasma sodium and potassium levels showed an unclear pattern for differing FMC concentrations and exposure times. Haematocrit values were affected by FMC treatment.     

水产品中的孔雀石绿残留及其研究概况

孔雀石绿,作为重要的工业原料和治疗水产动物水霉病的有效药物,近年来随着国际社会对食品安全卫生的重视以及其高残留、高毒性、致癌、致畸、致突变等毒副作用的披露,引起社会的广泛关注。本文详细介绍了孔雀石绿的性质、毒性、代谢和检测等方面的研究情况,旨在为孔雀石绿的监控工作提供相关信息,为监控工作提供依据。     

河蟹溞状幼体培育期使用孔雀石绿杀灭聚缩虫的试验

孔雀石绿对河蟹(氵蚤)状幼体的48小时半数致死浓度Ⅰ期幼体为1.89ppm,V期幼体为7.45ppm。在河蟹工厂化育苗中如果发生聚缩虫感染,可采用孔雀石绿1ppm浓度全池泼洒、10～12小时内排出药液水、技入新水的方法杀灭聚缩虫,不会影响河蟹(氵蚤)状幼体的变态发育。     

Towards strategic use of fungicides against Saprolegnia parasitica in salmonid fish hatcheries

Abstract. The life-cycle of Saprolegnia parasitica was considered in relation to the use of protective fungicides against it in salmonid fish hatcheries. Malachite green oxalate was the example fungicide, and all experiments were at 10°C. If the fungicide is to be used as a continuous addition to the water, as a protectant, then target stages of the fungus are identified as zoospores and secondary zoospore cysts: the latter, although physically sedimentary, have a remarkably high degree of buoyancy in the water and hence pose a constant threat to the fish. Continuous addition to the water, to give 0·25 mgl⁻¹ malachite green oxalate, would be expected to kill zoospores and zoospore cysts, but 0·05 mgl⁻¹, while allowing zoospore cyst germination, was a strong inhibitor to fungal growth and, aided by natural defences of the fish, might well succeed in disease prevention. If the fungicide is to be used as an intermittent addition to the water (every 24 h), then target stages of the fungus are small mycelia on the fish in addition to zoospores and zoospore cysts in the water. An experimental model was made to consider the vulnerability of these small mycelia, growing on the fish over the preceding 24 h, to malachite green oxalate. If mycelia are of narrow diameter, as expected, then 0·25 mgl⁻¹ malachite green oxalate, applied for 15 min, should control their growth and protect the fish.     

鳗鲡肌肉中孔雀石绿代谢物隐性孔雀石绿染料残留标准物质的研制

为建立鳗鲡肌肉中孔雀石绿代谢物隐性孔雀石绿染料残留标准物质的研制和定值方法,以一定质量浓度孔雀石绿对鳗鲡进行药浴给药,使孔雀石绿在鱼体内自然代谢,从而使鳗鲡体内含有隐性孔雀石绿残留。经均质、真空包装及辐照处理后,获得一批500个独立包装的的鳗鲡肌肉样本。采用超高效液相色谱-串联质谱法对该样本进行均匀性和稳定性检验,经8家独立实验室协同定值及不确定度评估,其特性值为2.82μg/kg,扩展不确定度为0.39μg/kg(k=2)。所建立的制备方法为染料残留鳗鲡基体标准物质的实验室制备提供了一种参考。     

小鳄龟纤毛虫病的防治研究

为筛选孔雀石绿的替代药物,探索小鳄龟纤毛虫病的有效治疗方法,比较了几种常见药物对小鳄龟纤毛虫病的治疗效果及对龟摄食和活动的影响。结果表明,高锰酸钾疗效最好,其次为氯化钠、甲醛溶液,聚维酮碘和硫酸铜无明显疗效;氯化钠和甲醛溶液对龟摄食、活动等影响较大。生产上可采用75g/m~3的高锰酸钾浸洗30分钟防治小鳄龟的纤毛虫病。     

三种水产药物对虹鳟幼鱼的急性毒性

本文研究了水产常规药物高锰酸钾、孔雀石绿、久效磷晶体敌百虫对虹鳟幼鱼的急性毒性。实验发现虹鳟幼鱼对高锰酸钾溶液24、48、96h的中间忍受限(TLM)分别为1.65、1.56和0.35mg/L,高锰酸钾对虹鳟幼鱼的安全浓度为0.035mg/L,虹鳟幼鱼对孔雀石绿溶液12、24、38、96h的中间忍受限分别为0.280、0.250、0.126、0.100mg/L。孔雀石绿对虹鳟幼鱼的安全浓度为0.010mg/L。虹鳟幼鱼对久效磷敌百虫中性溶液12、24、48、96h的中间忍受限分别为6.68、3.74、1.57、0.49mg/L,敌百虫对虹鳟幼鱼的安全浓度为0.049mg/L。     

Disrupting actions of bisphenol A and malachite green on growth hormone receptor gene expression and signal transduction in seabream

Environmental estrogen could mimic natural estrogens thereby disrupting the endocrine systems of human and animals. The actions of such endocrine disruptors have been studied mainly on reproduction and development. However, estrogen could also affect the somatotropic axis via multiple targets such as growth hormone (GH). In the present study, two endocrine disruptors were chosen to investigate their effects on the expression level and signal transduction of growth hormone receptor (GHR) in fish. Using real-time PCR, it was found that exposure to both the estrogenic (bisphenol A) and anti-estrogenic (malachite green) compounds could attenuate the expression levels of GHR1 and GHR2 in black seabream (Acanthopagrus schlegeli) hepatocytes. The expression level of IGF-I, the downstream effector of GHR activation in the liver, was decreased by bisphenol A but not by malachite green. Luciferase reporter assay of the β-casein promoter was used to monitor GHR signaling in transfected cells. In the fish liver cell line Hepa-T1, both GHR1 and GHR2 signaling were attenuated by bisphenol A and malachite green. This attenuation could only occur in the presence of estrogen receptor, indicating that these agents probably produce their actions via the estrogen receptor. Results of the present study demonstrated that estrogenic or anti-estrogenic compounds could down-regulate the somatotropic axis in fish by affecting both the gene expression and signaling of GHR. In view of the increasing prevalence of these compounds in the environment, the impact on fish growth and development both in the wild and in aquaculture would be considerable.