Field depuration and biotransformation of paralytic shellfish toxins in scallop<Emphasis Type="Italic"> Chlamys nobilis</Emphasis> and green-lipped mussel<Emphasis Type="Italic"> Perna viridis</Emphasis>

Under laboratory conditions, the scallop Chlamys nobilis and the mussel Perna viridis were exposed to N-sulfocarbamoyl toxins (C2 toxin), a paralytic shellfish toxin (PST), by feeding a local toxic strain of the dinoflagellate Alexandrium tamarense (ATDP) that produced C2 toxin exclusively. The bivalves were subsequently depurated in the field, and their depuration kinetics, biotransformation and toxin distribution were quantified. Depuration was characterized by a rapid loss within the first day, followed by a secondary slower loss of toxins. In the fast depuration phase, scallops detoxified PSTs more quickly than the mussels (depuration rate constants for scallops and mussels were 1.16 day^–1 and 0.87 day^–1, respectively). In contrast, the mussels detoxified PSTs more quickly than the scallops in the slow depuration phase, and the calculated depuration rate constants (mean+SE) from day 2 to day 13 were 0.063+0.009 day^–1 and 0.040+0.019 day^–1 for mussels and scallops, respectively. The differences in the appearances of gonyautoxins, GTX2 and GTX3, and their decarbamoyl derivatives, dcGTX2, dcGTX3 and GTX5, which are all derivatives of C2 toxin, indicated active and species-specific biotransformation of the algal toxins in the two bivalves. In both species of bivalves, the non-viscera tissue contained fewer toxins and lower concentrations than the viscera-containing tissue compartment. In scallops, very little toxin was distributed in the adductor muscle. In mussels, most of the PSTs were found in the digestive gland with significant transport of toxins into the digestive gland from other tissues during the course of depuration. The toxin profiles of scallops and mussels differed from each other and from that of the toxic algae fed. A significant fraction of GTX5 was detected in the mussels but not in the scallops. Our study demonstrates a species specificity in the depuration kinetics, biotransformation and tissue distribution of PSTs among different bivalves.Communicated by T. Ikeda, Hakodate     

Depuration of twelve trace metals in tissues of the oysters Crassostrea gigas and C. virginica

The depuration of 12 trace metals in the mantle, gill, digestive gland, and kidney of Crassostrea gigas and C. virginica was investigated under natural field conditions; oysters from a relatively contaminated environment (Redwood Creek in south San Francisco Bay) were transplanted to a relatively clean environment (Tomales Bay). In the transplanted oysters, the digestive gland and kidney depurated Cd, Cu, Hg, Ag, and Zn more readily than the mantle and gill. Other trace metals As, Fe, Mn, Ni and Se showed varying depuration patterns. The results for Cr and Pb were inconclusive, since initial concentrations were too low to follow any losses. Interspecific differences in trace metal depuration were observed. Biological half-lives for most trace metals were on the order of 23 to 60 d for C. gigas and on the order of 70 to 180 d for C. virginica.     

Characterisation and expression of four mRNA sequences encoding glutathione S-transferases pi,mu, omega and sigma classes in the Pacific oyster<Emphasis Type="Italic"> Crassostrea gigas</Emphasis> exposed to hydrocarbons and pesticides

Hydrocarbon and pesticide pollution in coastal ecosystems can disturb marine bivalve metabolism. In this study, we characterised four full-length cDNA sequences encoding glutathione S-transferases (GSTs) in the Pacific oyster Crassostrea gigas. A BLAST X search showed that these four sequences encode GSTs from four different classes: GST pi, sigma, mu and omega. A phylogenetic analysis of GST was made to determine the position of oyster GST compared to invertebrate and vertebrate sequences. We developed a semi-quantitative, multiplex RT-PCR to follow the expression of these four GSTs in tissues of oysters exposed to hydrocarbons and two pesticide treatments (glyphosate and a mixture composed of atrazine, diuron and isoproturon) under experimental conditions. Our results showed strong differential expression of these four GSTs that was both tissue specific as well as time and treatment dependent. We observed that expression levels were higher in digestive gland than in gill tissues in pesticide-exposed oysters. Furthermore, omega and mu class GST mRNA expression in the digestive gland might be useful as a possible marker of hydrocarbon exposure, while pi and sigma class GST mRNA expression in the digestive gland may be similarly useful as a marker of pesticide exposure in monitoring programmes.Communicated by S.A. Poulet, Roscoff     

Accumulation,tissue distribution and elimination of 203HgCl2 and CH3 203HgCl in the tissues of the American oyster Crassostrea virginica

Oysters (Crassostrea virginica) were exposed for 3 days to mercury-203 labeled HgCl₂ or CH₃HgCl added directly to artificial seawater or added preconcentrated on the marine diatom Phaeodactylum tricornutum. The concentration of mercury in 5 tissues was measured for 45 days after mercury was removed from the ambient water. At the beginning of the depuration period, the highest concentrations of mercury in tissues were attained in: gill>digestive system>mantle>gonad>muscle in oysters exposed to water containing mercury; and in digestive system>gill>mantle> gonad>muscle in oysters fed labeled algae. This same distribution pattern is seen for both chemical forms of mercury. Although the initial pattern of accumulation was identical for both mercury compounds within each exposure group, the fate of the accumulated mercury was very different after 45-days depuration. In oysters accumulating mercury directly from seawater, inorganic mercury residues rapidly declined in gill and digestive tissue, but were slowly reduced in mantle, gonadal, and muscle tissue. This pattern was duplicated by oysters exposed to methyl mercuric chloride in seawater except that gonadal and muscle residues greatly increased during depuration. In oysters ingesting labeled P. tricornutum cells, mercuric chloride and methyl mercuric chloride residues rapidly declined in gill and digestive tissue, remained constant in the mantle, but sharply increased in gonadal and muscle tissue during depuration.     

How do slugs cope with toxic alkaloids?

Summary. Arion lusitanicus and other slugs are able to feed on a variety of plants, even those containing toxic secondary metabolites, such as alkaloids. Alkaloids, like sparteine, lupanine, quinidine and atropine are avoided in no-choice feeding experiments only when they are present in concentrations above 1 mg/g food pellet. Alkaloids (sparteine, lupanine, cytisine, quinidine, atropine, senecionine, eserine, and harmaline) are toxic to Arion lusitanicus when injected. LD₅₀ values are 10 to 20 fold higher than in vertebrates indicating that slugs have a high tolerance towards food toxins. The tolerance is higher in young animals than in adult slugs. Injected alkaloids are rapidly detoxified within 72 h. Tolerance and detoxification can be induced by feeding slugs on non-lethal doses of lupin alkaloids. Using isolated microsomal preparations from the digestive gland, active detoxification was observed in vitro. Evidence is presented that cytochrome p450 plays an important role in detoxification of the applied alkaloids. A powerful and inducible detoxification systems appears to be the main mechanism that allows slugs to feed on plants rich in secondary metabolites (when no other food is available), that are usually avoided by other herbivores.     

Effect of sexual maturation on the tissue biochemical composition of<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> and<Emphasis Type="Italic"> O</Emphasis>.<Emphasis Type="Italic"> defilippi</Emphasis> (Mollusca: Cephalopoda)

Changes in the protein, lipid, glycogen, cholesterol and energy contents, total amino acid and fatty acid profiles of Octopus vulgaris and O. defilippi tissues (gonad, digestive gland and muscle) during sexual maturation (spermatogenesis and oogenesis) were investigated. Both species showed an increase of amino acids and protein content in the gonad throughout sexual maturation (namely in oogenesis), but allocation of these nitrogen compounds from the digestive gland and muscle was not evident. The major essential amino acids in the three tissues were leucine, lysine and arginine. The major non-essential amino acids were glutamic acid, aspartic acid and alanine. With respect to carbon compounds, a significant increasing trend (P<0.05) in the lipid and fatty acid contents in the three tissues was observed, and, consequently, there was also little evidence of accumulated lipid storage reserves being used for egg production. It seems that for egg production both Octopus species use energy directly from food, rather than from stored products. This direct acquisition model contrasts with the previous model for Octopus vulgaris proposed by ODor and Wells (1978: J Exp Biol 77:15–31). Most of saturated fatty acid content of the three tissues was presented as 16:0 and 18:0, monounsaturated fatty acid content as 18:1 and 20:1 and polyunsaturated fatty acid content as arachidonic acid (20:4n-6), eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3). Though cholesterol is an important precursor of steroid hormones, this sterol content exhibited variations that do not seem to be related with the maturation process. Moreover, significant differences (P<0.05) were obtained between genders, suggesting that perhaps there is a greater physiological demand for cholesterol during spermatogenesis than oogenesis. If the component sterols of octopus are of a dietary origin, considerable variation in the cholesterol content between species might be expected on the basis of the sterol composition of their prey. The glycogen reserves increased significantly in the gonad and decreased significantly (P<0.05) in the digestive gland and muscle of O. vulgaris (these trends were not evident in O. defilippi). Glycogen may play an important role in the maturation process and embryogenesis of these organisms, because carbohydrates are precursors of metabolic intermediates in the production of energy. It was evident that sexual maturation had a significant effect upon the gonad energy content, but the non-significant energy variation (P>0.05) in the digestive gland and muscle revealed no evidence that storage reserves are transferred from tissue to tissue. The biochemical composition of digestive gland and muscle may not be influenced by sexual maturation, but rather by other biotic factors, such as feeding activity, food availability, spawning and brooding.Communicated by S.A. Poulet, Roscoff     

Effects of lead exposure on redox status,DNA and histological structures in Venus verrucosa gills and digestive gland

ABSTRACT

Lead (Pb) is one of the most toxic heavy metals that affect the physiological status of aquatic organisms. The present investigation evaluated the possible toxic effect of lead chloride (PbCl₂) on biomarkers responses, DNA damage and histological alterations in Venus verrucosa gills and digestive gland. Three concentrations of PbCl₂ (D1:1µgL^?1, D2: 10µgL^?1 and D3: 100µgL^?1) were chosen for V. verrucosa exposure during six days. At the end of the trial, it was found that Pb tended to accumulate in both gills and digestive gland in a dose-dependent manner. However, gill tissues exhibited the highest metal burden. Our results showed an increase of malondialdehyde, protein carbonyls and advanced oxidation protein product levels in both organs following PbCl₂exposure. The induction of both non-enzymatic and enzymatic antioxidant systems; as well as the decrease of the acetylcholinesterase activity and degradation of DNA structure was recorded in the gills and digestive gland. The histopathological alterations observed in gills (disruption of lamellas and cilia filaments?…) and digestive gland (lumens occlusion, necrosis and fibrosis) confirmed the aforementioned results. Our data highlighted the short-term toxicity effects of PbCl₂ on V. verrucosa and pointed out a high sensitivity of gills towards this metal.     

Copper sulphate (CuSO4) toxicity on tissue phosphatases activity and carbohydrates turnover in Achatina fulica

A time course study on the sublethal toxicity of CuSO4 on tissue carbohydrate metabolites level and their phosphatases activity in Achatina fulica revealed differential response. The levels of total carbohydrates and glycogen in the body mass muscle, foot muscle and hemolymph revealed their involvement in the endogenous derivation of energy during stress. The same metabolites in digestive gland revealed its importance to reproduction and development. The lactate accumulated in all the tissues implied the mechanism of CuSO4 toxicosis in the metabolic acidosis. The decrease of pyruvate in foot muscle, body mass muscle and hemolymph inferred the preponderance of glycolysis in energy derivation. In contrast, the pyruvate concentration in digestive gland revealed its differential response in the stress metabolic sequence of changes, as a unique tissue. The lactate/pyruvate ratio and the calcium content in tissues constitute direct evidences for the snails adaptation to toxic stress.     

Tissue distribution of the amnesic shellfish toxin,domoic acid,in<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> from the Portuguese coast

Domoic acid (DA), the amnesic shellfish toxin, is a food-web-transferred algal toxin that has been detected in many marine organisms from copepods to whales. However, cephalopods, which are important members of the food chain, have never been implicated in DA transfer or accumulation. Here, we present data showing relevant values of DA detected in the common octopus (Octopus vulgaris) from the Portuguese continental coast. Even though DA is hydrophilic and is not expected to be accumulated in the tissues, DA was always detected in our octopus tissue samples. Tissue distribution of DA revealed that the digestive gland and the branchial hearts are the main organs of DA accumulation. Highly variable DA concentrations, ranging from 1.1 to 166.2 g DA g^–1, were observed in the digestive glands. Low levels of DA were detected in the digestive tract (stomach and intestine) and could be a consequence of high digestion rates or a result of non-exposure to toxic vectors during the sampling period. In fact, octopus prey, such as bivalves, crustaceans and fishes, are known to occasionally work as DA vectors. Consequently, DA uptake into octopus tissues is likely sporadic. Similar low levels were detected in the kidney, gills, systemic heart, posterior salivary glands and mantle, and no DA was found in either the gonads or the ink sac. These data are the necessary first step towards achieving an understanding of the accumulation of phycotoxins in O. vulgaris.Communicated by S.A. Poulet, Roscoff     

有机磷农药对缢蛏[Sinonovacula constricta (Lamarck)]显微结构的影响

在实验室模拟缢蛏的生长环境,研究乐果和三唑磷对缢蛏的毒性效应.以气相色谱法测定缢蛏体内有机磷农药的含量,并通过光镜观察有机磷农药对缢蛏鳃和消化腺显微结构的影响.结果表明:缢蛏体内乐果和三唑磷的含量随着水体中有机磷农药的增加而增加,有很好的剂量-效应关系,但是无线性相关性.乐果和三唑磷对缢蛏鳃显微结构的影响是:鳃丝变短,鳃丝间隔和基底膜结缔组织增生,鳃孔、血管腔堵塞;不同的是:在乐果胁迫下,上皮细胞纤毛脱落严重,而三唑磷胁迫下,上皮细胞纤毛增生.乐果和三唑磷对缢蛏消化腺显微结构的影响主要表现为腺细胞界限模糊,腺泡水肿变形、部分坏死;结缔组织增生,细胞间质疏松,出现许多空泡;有机磷农药的浓度高时消化腺细胞变性、坏死,细胞质囊泡化.     

纳米氧化镍颗粒对长牡蛎(Crassostreagigas)抗氧化防御体系的影响

纳米氧化镍(nNiO)作为一种广泛使用的纳米颗粒,其水生毒理效应研究还很有限。为探索n Ni O对海洋贝类的毒性机制,本研究将长牡蛎(Crassostrea gigas)置于不同浓度(0、1、10、100 mg·L~(-1))的n Ni O中暴露96 h,分别测定鳃和消化腺组织的丙二醛(MDA)含量和超氧化物歧化酶(SOD)、过氧化物酶(POD)以及过氧化氢酶(CAT)活性,并通过实时荧光定量PCR技术测定了鳃和消化腺中应激蛋白HSP70和AOX基因的表达变化。结果显示:在100 mg·L~(-1)n Ni O处理下,2种组织中MDA含量均显著性升高(P0.01),显示纳米颗粒造成了长牡蛎的脂质过氧化,并可能引起相应的氧化损伤。同时,n Ni O暴露也诱导了长牡蛎抗氧化酶(SOD、CAT和POD)活性的改变。其中,SOD和CAT活性在10 mg·L~(-1)浓度处理组达到最高,而POD活性在1 mg·L~(-1)浓度组即达最高值。在高浓度n Ni O(100 mg·L~(-1))胁迫下,3种抗氧化酶的活性均比低浓度(1和10 mg·L~(-1))处理组降低,表明抗氧化酶的保护作用在较低浓度暴露下更有效;而热激蛋白(hsp70)和交替氧化酶(aox)基因却分别在长牡蛎消化腺和鳃组织中上调表达(P0.01),并表现出一定的组织差异。说明高浓度纳米颗粒暴露中主要是应激蛋白发挥了作用。本文结果为纳米氧化镍对海洋双壳贝类的毒性机制研究及生态风险评估提供了基础数据。     

The effect of copper and zinc on the metabolism of the mussel Mytilus edulis

The effects of copper and zinc on the metabolism of the mussel Mytilus edulis (L.) and its component tissues were studied. 500 ppm copper sodium citrate inhibited oxygen consumption of the whole animal and gill tissue, but no similar effect was observed on digestive gland tissue. 500 ppm zinc sodium citrate exerted no effect upon gill or digestive gland respiration, and neither metal salt affected the respiration of homogenates of gill, digestive gland or gonad. Direct observation of gill tissues during exposure to the metals revealed that 500 ppm copper sodium citrate caused inhibition of ciliary activity; exposure of tissues to 2 ppm Cu for 24 h resulted in only partial inhibition of the cilía. It is suggested that metabolic suppression noted in whole animals and gill tissues is due to the inhibition of an energy-consuming process such as ciliary activity rather than interference with respiratory enzyme systems.     

Hydrocarbon pollution of edible shellfish by an oil spill

A spill of 650,000 to 700,000 l of No. 2 fuel oil has contaminated the coastal areas of Buzzards Bay, Massachusetts (USA). Gas chromatography demonstrates the presence of this oil in the sediments of the affected area. Two months after the accident, essentially unchanged oil is still being released from the sediments. The presence of the same pollutant is demonstrated in whole oysters Crassostrea virginica and in the adductor muscle of the scallop Aequipecten irradians. A presumably biochemical modification leads to a gradual depletion of the straight chain and, to a lesser extent, of branched chain hydrocarbons. This does not result in detoxification, as the more toxic aromatic hydrocarbons are retained in the organisms several months after the accident. Scallops from an uncontaminated area contain hydrocarbons in lesser amounts and of very different molecular weight and type distribution; they are accountable entirely from biological sources.Contribution No. 2444 of the Woods Hole Oceanographic Institution.     

Effects of induced paralysis on hemocytes and tissues of the giant lions-paw scallop by paralyzing shellfish poison

In general, bivalves are not affected by exposure to toxic dinoflagellates that produce paralyzing shellfish poisons (PSP). After injection with PSP extracted from the Gymnodinium catenatum, Nodipecten subnodosus is paralyzed, indicating that PSP provokes effects similar to what is observed in vertebrates, including paralysis and metabolic stress. To investigate the processes involved in poisoning by PSP, lions-paw scallops were injected with gonyautoxin (GTX) 2/3 epimers in the adductor muscle. Mild doses provoked adductor muscle contractions and paralysis, mantle retraction, and incapacity of shell closure, but scallops gradually recovered in a clear, dose-time recovery pattern. With high doses of GTX 2/3, scallops were permanently paralyzed, and hemocytes in hemolymph were reduced. Surprisingly, under these conditions, scallops continued normal feeding and did not show any microscopic defect in intestine or gills, but hemocytes infiltrated the adductor muscle and abnormal vitellogenesis and mantle melanization occurred. Paralysis stress was accompanied by negative scallop responses, based on visible effects, generation of nitric oxide, lipid peroxidation, and changes in antioxidant and hydrolytic enzymes in hemocytes and tissues. These data can be used to understand potential side effects of PSP in bivalves.     

Concentration and distribution of heavy metals in tissues of two cephalopods,Eledone cirrhosa and Sepia officinalis,from the French coast of the English Channel

The concentrations of 11 heavy metals (Ag, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, V and Zn) were measured in the tissues (digestive gland, branchial hearts, gills, digestive tract, kidney, genital tract, muscle, skin, shell) of the two cephalopods Eledone cirrhosa (d'Orb.) and Sepia officinalis (L.) collected from the French coast of the English Channel in October 1987. The tissues of both species displayed a similar pattern of heavy-metal accumulation: the digestive gland, branchial hearts and kidney were the major sites of concentration for all 11 metals; the digestive gland accumulated silver, cadmium, cobalt, copper, iron, lead and zinc, the branchial hearts high concentrations of copper, nickel and vanadium, and the kidney high concentrations of manganese, nickel and lead. The digestive gland, which constituted 6 to 10% of the whole-animal tissue, contained >80% of the total body burden of Ag, Cd and Co and from 40 to 80% of the total body burden of the other metals. The ratios between heavy metal concentrations in the digestive gland and those in the muscle separated the elements into three groups, those with a ratio 10 (Cr, Mn, Ni, Pb, V, Zn), those with a ratio >10 to <50 (Co, Cu, Fe), and those with a ratio 50 (Ag, Cd). The digestive gland of cephalopods (carnivorous molluscs whose age can be easily calculated with great accuracy) would seem to constitute a good potential indicator of heavy metal concentrations in the marine environment.     

虾夷扇贝毒素(yessotoxin)诱导的栉孔扇贝(Chlamys farreri)体内P糖蛋白转运活性及其分布研究

大量研究表明P-糖蛋白(P-glycoprotein,P-gp)介导的多型异源物质抗性机制(multi-xentiobiotic resistance,MXR)在贝类耐受或抵抗污染水域中有毒有害物质的毒害中具有重要作用,但是该机制在贝类抵抗或耐受食源性毒素方面作用的研究却很少。本研究以栉孔扇贝(Chlamys farreri)为实验动物,向其投喂产虾夷扇贝毒素(yessotoxins,YTXs)的网状原角藻(Protoceratium reticulatum),通过检测栉孔扇贝鳃和消化腺组织中YTX毒素含量随时间的变化情况,结合2种组织中P-gp介导的MXR活性变化及P-gp在组织中的表达和分布,探讨P-gp在扇贝耐受YTX毒素中可能的作用。研究结果表明,随着毒藻暴露时间的延长,栉孔扇贝鳃和消化腺中YTX含量逐渐增加,且具有明显的时间依赖关系。YTX的大量累积虽未导致扇贝死亡,但是仍然对扇贝组织产生不同程度的毒性。苏木精-伊红(hematoxylin-eosin,HE)染色发现,染毒12 h后,栉孔扇贝鳃组织前端出现明显黑化,消化腺组织导管上皮细胞部分脱落入管腔中,腺管内细胞界限模糊,部分腺管崩解。罗丹明B(rhodamine B,Rho B)外排实验表明YTX毒素暴露能显著增强栉孔扇贝鳃和消化腺组织中P-gp的转运活性,暴露12 h后鳃和消化腺Rho B的排出速率就已分别达到对照组的3.8倍和1.4倍,随着暴露时间的延长,P-gp活性略有增强,72 h后开始下降;P-gp的功能性抑制剂维拉帕米(verapamil,VRP)在实验设定的浓度下没有抑制反而增强了Rho B的外排;免疫定位显示,毒素暴露后栉孔扇贝鳃丝前端纤毛柱状细胞及消化腺导管上皮柱状细胞和腺管消化细胞P-gp免疫阳性信号增强,说明YTX毒素暴露提高了P-gp的表达量。综上推断,YTX毒素可能是P-gp的底物,它能够诱导P-gp转运活性增强,并增加P-gp的表达量来参与扇贝对YTX毒素的耐受或抗性;设定浓度的VRP不能抑制P-gp的活性,这可能是由于种属差异造成的,关于VRP对栉孔扇贝Pgp的作用还有待进一步的研究。     

Geese and dietary allelochemicals — food palatability and geophagy

Summary The palatability of plants was studied in young, immature and adult geese of three species,Anser anser, Anser indicus andBranta canadensis, with respect to secondary plant metabolites. In their first 1–4 weeks of life, hand-reared goslings feed on a wide variety of plants, more or less irrespective of their allelochemical contents. Older birds become more selective, but still consume plants which are normally considered to be unpalatable or even toxic for other animals. Choice experiments were performed with pure secondary metabolites which were offered on otherwise highly palatable food items, such as leaves ofTaraxacum officinale. These experiments revealed a similar trend, in that very young goslings discriminate their food much less than older goslings or adult geese. In general, food contaminated with essential oils was rejected, whereas alkaloids, glycosides, amines and sulfur compounds were tolerated to a remarkable degree. In consequence, especially young, but also adult geese must have a high capacity to tolerate and/or to detoxify dietary allelochemicals. Another detoxification mechanism became evident during the experiments: geese ingest soil and mud quite regularly. It could be shown experimentally that the respective soil had a high capacity to bind alkaloids. We assume that geophagy is a means (besides a presumed active detoxification in the liver) to adsorb and thus reduce the contents of dietary allelochemicals.     

Role of plant growth promoting rhizobacteria in the remediation of metal contaminated soils

Pollution of the biosphere by the toxic metals is a global threat that has accelerated dramatically since the beginning of industrial revolution. The primary source of this pollution includes the industrial operations such as mining, smelting, metal forging, combustion of fossil fuels and sewage sludge application in agronomic practices. The metals released from these sources accumulate in soil and in turn, adversely affect the microbial population density and physico-chemical properties of soils, leading to the loss of soil fertility and yield of crops. The heavy metals in general cannot be biologically degraded to more or less toxic products and hence, persist in the environment. Conventional methods used for metal detoxification produce large quantities of toxic products and are cost-effective. The advent of bioremediation technology has provided an alternative to conventional methods for remediating the metal-poisoned soils. In metal-contaminated soils, the natural role of metal-tolerant plant growth promoting rhizobacteria in maintaining soil fertility is more important than in conventional agriculture, where greater use of agrochemicals minimize their significance. Besides their role in metal detoxification/removal, rhizobacteria also promote the growth of plants by other mechanisms such as production of growth promoting substances and siderophores. Phytoremediation is another emerging low-cost in situ technology employed to remove pollutants from the contaminated soils. The efficiency of phytoremediation can be enhanced by the judicious and careful application of appropriate heavy-metal tolerant, plant growth promoting rhizobacteria including symbiotic nitrogen-fixing organisms. This review presents the results of studies on the recent developments in the utilization of plant growth promoting rhizobacteria for direct application in soils contaminated with heavy metals under a wide range of agro-ecological conditions with a view to restore contaminated soils and consequently, promote crop productivity in metal-polluted soils across the globe and their significance in phytoremediation.     

Paralytic shellfish poison in the “hiogi” scallop Chlamys nobilis

Attempts were made to analyze the toxin composition of the toxic hiogi scallop Chlamys nobilis. The toxins were partially purified from the digestive glands by column chromatography using Bio-Gel P-2 and Bio-Rex 70 (H⁺ form), resulting in separation into protogonyautoxin (PX), gonyautoxin (GTX) and saxitoxin (STX) fractions. Their total potencies were scored to be <100 mouse units (MU), 3200 MU and 3700 MU, respectively. A 5-min hydrolysis with 0.1 N HCl enhanced the potencies of PX and GTX fractions to 450 MU and 12000 MU respectively, whereas no enhancement occurred in the STX fraction at all. Electrophoretic, thin-layer chromatographic and high performance liquid chromatographic analyses demonstrated that the PX fraction consisted mainly of GTX₈ and its epimer, the GTX fraction of GTX₅, GTX₆, along with two unidentified toxins, and the STX fraction exclusively of two unidentified toxins. This rather unique composition suggested a complex metabolism of PSP in this species.     

Effects of the toxic dinoflagellate <Emphasis Type="Italic">Gymnodinium catenatum</Emphasis> on uptake and fate of paralytic shellfish poisons in the Pacific giant lions-paw scallop <Emphasis Type="Italic">Nodipecten subnodosus</Emphasis>

Juvenile Pacific giant lions-paw scallops Nodipecten subnodosus were fed the toxic dinoflagellate Gymnodinium catenatum, a producer of paralytic shellfish poison (PSP), supplied with Isochrysis galbana (a nontoxic microalgae). Short-term (<24 h) experiments were performed to determine clearance and ingestion rates of G. catenatum. Kinetics of PSP was examined in longer-term experiments (>2 days). At high food concentrations, juvenile scallops showed production of pseudofeces, partial shell valve closure, and reduction in feeding. According to HPLC analysis, the only toxins present in the dinoflagellate G. catenatum and in the scallops were the gonyautoxins (GTXs), except in the labial palps and digestive gland, where trace amounts of saxitoxin (STX) were present in scallops. These tissues could play an important role in toxin biotransformation. The ranking of toxin concentration in tissues was: digestive gland > labial palps > intestine > gills > mantle > adductor muscle, where the total contribution of viscera was more than 80% of the total toxin body burden. Juvenile scallops exhibited no apparent detrimental physiological responses during the long-term feeding experiment. The dinoflagellate may contribute nutrients to the scallop, in addition to the microalgae I. galbana. The dinoflagellate may enhance cell uptake and byssus production. Once PSP accumulated during the first 12 days, it was slowly eliminated. The limited capacity for accumulating toxins in the adductor muscle favors domestic marketing of scallops.