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1.
The stability of lysosomes in the digestive cells of Mytilus edulis is affected by changes in both the chemical and physical conditions of the environment.1 This type of response must reflect, at least in part, some change in the organisation of the lysosomal membrane, which is a structure which can be resolved by the electron microscope. However, conventional methods of tissue preparation have produced low contrast images of the digestive gland and damage to the lysosomal membranes. Recently a cryopreparation technique2 has produced lysosomal membrane preparations in the digestive cells of the digestive gland of Mytilus which have high contrast and structural integrity (Fig. 1) in control animals. This method has been used to demonstrate pathological alterations in the lysosomal membrane induced by the polynuclear aromatic hydrocarbon (PNAH) phenanthrene. Anthracene, an isomeric PNAH, had no effect on the morphology of the lysosomal membrane. Phenanthrene also induced apparent proliferation of smooth endoplasmic reticulum in the digestive cells. 相似文献
2.
Gorbi S Virno Lamberti C Notti A Benedetti M Fattorini D Moltedo G Regoli F 《Marine environmental research》2008,65(1):34-49
An ecotoxicological protocol with caged mussels, Mytilus galloprovincialis, was developed to evaluate the potential impact of an offshore gas platform in the central Adriatic Sea. Reference organisms were collected on a seasonal basis from an unpolluted site and transplanted for four weeks in both the sampling area and to the investigated platform. Chemical analyses of trace metals in mussel tissues were integrated with a multi-biomarker approach for the early detection of biological responses at several cellular targets. Induction of metallothioneins, peroxisomal proliferation and activity of acetylcholinesterase were measured as markers for specific classes of chemicals. Special attention was given to oxyradical metabolism and appearance of oxidative-mediated toxicity to reveal a more general onset of cellular disturbance. In addition to individual antioxidants (superoxide dismutase, catalase, glutathione S-transferases, glutathione reductase, Se-dependent and Se-independent glutathione peroxidases, and levels of total glutathione), the total oxyradical scavenging capacity (TOSC) allowed a quantification of the overall capability to neutralize specific forms of intracellular reactive oxygen species (ROS; i.e. peroxyl and hydroxyl radicals). Cellular damages were evaluated as lysosomal destabilization (membrane stability, accumulation of lipofuscin and neutral lipids), lipid peroxidation products (malondialdehyde) and DNA integrity (strand breaks and micronuclei); the air survival test was finally applied to evaluate the overall physiological condition of mussels. Concentration of trace metals (As, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Zn) revealed only limited variations in transplanted mussels during various experimental periods and such changes appeared partly related to natural fluctuations. Among biological responses, variations of antioxidants and lysosomal stability were confirmed as sensitive early warning signals for biological disturbance of both natural and anthropogenic origin. The presented protocol with caged mussels allowed marked biological effects caused by the investigated platform to be excluded, and represented a useful approach that is easy to extend for monitoring the impact of offshore activities in the Adriatic sea. 相似文献
3.
Effects of temperature on scope for growth and accumulation of Cd, Co, Cu and Pb by the marine bivalve Mytilus edulis 总被引：7，自引：0，他引：7
In many aquatic organisms including Mytilus edulis, the role of temperature on bioaccumulation of metals is still not clearly understood. In this study, uptake and accumulation of Cu, Co, Cd and Pb in mussels were investigated at different temperatures (6-26 degrees C). Results from exposure of isolated gills showed a positive relationship between temperature and metal uptake. But in whole organism experiments, only the accumulations of non-essential metals (Cd, Pb) showed a similar trend while the two essential metals Co and Cu were independent and inversely related to temperature, respectively. With exception of Cu, elimination process appeared to be independent of temperature. The study also showed that neither changes in scope for growth (SFG) of mussels nor chemical speciation could fully account for the observed temperature-effects. Overall, these results suggest that fundamentally (i.e. at epithelial membranes), temperature-effects on uptake are largely due to changes in solution chemistry and physical kinetics, which favours higher uptake at high temperature. But at whole organism level, complex physiological responses appears to mask the relationship, particularly for biologically essential metals like copper. 相似文献
4.
Lysosomal sequestration of polynuclear aromatic hydrocarbons (PNAHs), a major class of environmental contaminant, is a well-established phenomenon;1 considerably less is known about their pathological effects on lysosomes. Marine molluscs contain a number of lysosome-rich tissues and PNAHs are known to induce deleterious alterations in lysosomal structure and latency of lysosomal enzymes.2 The latter are presumed to involve destabilisation of the lysosomal membrane, resulting in increased permeability and reduced enzyme latency. If lysosomal injury involves derangement of membrane-lipid structure due to the interaction of PNAHs then it would be expected that membrane damage would be closely linked to the structural characteristics of the intruding molecule. Our results show that the effects of the isomeric PNAHs phenanthrene and anthracene on digestive cell lysosomal stability were markedly different in the marine mussel (Mytilus edulis) over the same range of tissue concentrations. Lysosomal membrane stability was determined using a cytochemical test for enzyme latency.3 相似文献
5.
Certain specific aspects of cellular structure-linked functions can be used as rapid and sensitive indicators of cellular responses to environmental stimuli such as chemical contaminants.1 It is possible to observe structural-functional alterations in lysosomal membrane stability in the cells of some marine organisms at an early stage of such a response to cell injury,1,2 thus providing an early-warning signal of a potentially deleterious environmental situation. In order to further investigate lysosomal membrane disturbances, mechanisms of injury and capacity for cellular regeneration, the marine snail Littorina littorea was exposed to phenanthrene, which was used as an environmentally widespread and representative polynuclear aromatic hydrocarbon. Lysosomal membrane stability was measured using cytochemical determination of hydrolase latency,2 and lysosomal lipofuscin was measured as a possible indicator of enhanced lipid peroxidative damage.3 相似文献
6.
A field study was carried out in the Lagoon of Venice (north-east Italy) with the aim of evaluating the potential use of lysosomal destabilisation as a biomarker of anthropogenic stress in the autochthonous mussel Mytilus galloprovincialis. Two different approaches were adopted in biomonitoring six sites in the Lagoon, evaluating indigenous populations of mussels and organisms transplanted from a reference site and checked at several points in time. Lysosomal membrane stability was investigated by means of two tests: neutral red retention assay (NRRA) for evaluating haemocyte lysosomes and lysosomal latency test (LLT) for digestive cell lysosomes. Results indicate that the lysosomal response measured in haemocytes according to NRRA is a more valuable biomarker of anthropogenic stress in the framework both of passive and active biomonitoring in marine coastal environments. 相似文献
7.
This study was conducted to investigate the adaptability of marine mussels Mytilus edulis of increasing age to induced stress and subsequent recovery. Lysosomes, present in large numbers in the molluscan digestive gland, play a major role in intracellular digestion, and the stability of their membranes provides a sensitive biomarker for generalised cell injury which is correlated with the stress response of the whole animal. Lysosomal stability was measured in mussels of three age groups (2–4, 6–8 and ≥ 10 years) during exposure to hypoxia/hyperthermia and, in a separate experiment, to copper (50 ppb: where billion = 109). The lysosomal reactions of all three age groups to both experimental stressors were similar. However, recovery from the induced pathological reactions was most pronounced in the youngest animals and least apparent in the oldest group. These findings indicate that the stress reaction is independent of age but that the potential for recovery of lysosomal integrity is age-related. 相似文献
8.
网箱养殖大黄鱼越冬期间脂肪酸的相对含量变化 总被引：1，自引：0，他引：1
利用气-质法(GC-MS)对越冬期间海水平均温度在20,16,12,10和8 ℃条件下的2龄大黄鱼肌肉和肝脏组织中脂肪酸组成及相对含量进行分析,结果表明:随着海水水温的降低,机体会增加不饱和脂肪酸含量,尤其是EPA[C20:5(n-3)],EPA在肌肉中的含量从20 ℃时5.71%增至8 ℃时8.06%,在肝脏中从20 ℃时6.76%增至8 ℃时8.44%。由此可见,在越冬禁食条件下,大黄鱼机体通过自身的脂肪酸动态转化和代谢,减少饱和脂肪酸含量,而增加不饱和脂肪酸含量,以此增加膜的流动性,抵御寒冷。 相似文献
9.
Protein expression and lysosomal stability were observed in Mytilus edulis exposed to 0, 20, 40, 60, and 80 ppb copper sulfate for 24 h in a static exposure system. Haemolymph was removed from each mussel and analyzed for lysosomal damage using the Neutral Red retention assay expressed as the ratio of lysosomal size to cell area, the lysosomal destabilization ratio. Gill tissue from the same individuals was dissected, homogenized and the proteins extracted and then separated using two-dimensional gel electrophoresis. Two-dimensional protein maps from each concentration were analyzed using MELANIE II 2D gel analysis software (Biorad). Lysosomal damage increased with each dosage, with LDRs ranging from 0.18 +/- 0.01 at 0 ppb copper sulfate to 0.32 +/- 0.11 at 80 ppb copper sulfate. Orthogonal comparisons showed that the control, 20 and 40 ppb classes together differed significantly (P < 0.05) from the 60 and 80 ppb classes together. Specific protein expression signatures were identified at each dosage. 相似文献
10.
A Viarengo M Lafaurie G P Gabrielides R Fabbri A Marro M Roméo 《Marine environmental research》2000,49(1):1-18
The results of an intercalibration exercise among the laboratories participating in the MED POL program for monitoring biological effects of pollutants along the Mediterranean coasts are presented. Three established biomarkers, i.e. lysosomal membrane stability, metallothionein concentration and ethoxyresorufin-O-deethylase (EROD) activity, were intercalibrated. The stability of lysosomal membranes in mussels (Mytilus galloprovincialis Lam.) was assessed with a cytochemical method. The four participating laboratories were able to discriminate between control animals (membrane labilization times ranging from 21 to 35 min) and Cu-exposed animals (40 micrograms/l Cu for 3 days) (labilization times ranging from 4.5 to 7.4 min). The metallothionein concentration was evaluated in digestive gland homogenates of control mussels and of animals exposed to 200 micrograms/l Cd for 7 days. The eight participating laboratories were able to discriminate between controls and treated samples using a spectrophotometric method. The EROD activity was evaluated by 11 laboratories. All laboratories were able to discriminate between liver microsomal preparations obtained from control and from benzo-a-pyrene exposed fish (Dicentrarchus labrax), with values ranging from 0.5 to 15.88 pmol/min/mg protein in controls and from 5.41 to 165.13 pmol/min/mg protein in treated animals. Using S9 fractions, it was possible to correctly identify control and treated fish, with a variation similar to that found using microsomal fractions, albeit with an inevitable difference in specific activity. As a corollary, all laboratories involved produced comparable data and were able to identify pollutant-induced stress syndromes in sentinel organisms. Thus, intercalibration enables the use of biomarkers in large biomonitoring programs. 相似文献
11.
M.N. Moore J. Widdows J.J. Cleary R.K. Pipe P.N. Salkeld P. Donkin S.V. Farrar S.V. Evans P.E. Thomson 《Marine environmental research》1984,14(1-4)
Most investigations of the responses of marine organisms to xenobiotics have concentrated on single contaminants and little is known of possible interactive effects of different classes of xenobiotics. As these latter seldom occur in environmental isolation, it is important to understand any interactions (synergistic or antagonistic) which may occur. This problem has been approached in the mussel Mytilus edulis by exposing estuarine mussels to copper (20 μg litre−1) and phenanthrene (100 μg litre−1) both individually and in combination, and measuring cytochemical subcellular and physiological responses after 3 days exposure and 3 days and 12 days recovery period. Results showed that mussels accumulated both xenobiotics during 3 days exposure. Depuration of copper was complete in 3 days recovery period, while loss of phenanthrene ranged from 30% to 70% of the concentration reached after 3 days exposure. There were no interactive effects on depuration.Both copper and phenanthrene reduced lysosomal hydrolase latency in digestive cells, and copper appeared to have a synergistic effect in preventing recovery of latency of lysosomal N-acetyl-β-hexosaminidase during the recovery period. There was evidence, in the digestive cells, of an antagonistic effect of copper on stimulation of activity of the microsomal respiratory chain (measured as NADPH-neotetrazolium reductase) by phenanthrene. Stimulation of this system by phenanthrene persisted after 12 days recovery period. There was a synergistic interaction of copper and phenanthrene on elevation of oxygen consumption and ammonium excretion. Clearance rates and scope for growth (physiological condition) were depressed by copper but not by phenanthrene after 3 days exposure.These findings are discussed in terms of known effects of copper and phenanthrene and the interactions are considered in terms of environmental effects measurements. 相似文献
12.
A. Viarengo M.N. Moore G. Mancinelli A. Mazzucotelli R.K. Pipe 《Marine environmental research》1985,17(2-4)
Metallothioneins and lysosomes are known to be involved in cellular detoxication and sequestration of certain metals1–3 and both have been identified in this role in elimination of copper from marine mussels (Mytilus edulis/galloprovincialis).3 Cadmium (Cd), however, has been shown to persist in the cells of the digestive gland for long periods with only minimal elimination. An experiment was designed to test the effects of Cd on the fragility of lysosomal membranes in the digestive cells as a measure of cellular injury,4,5 metallothionein content of the digestive gland and cadmium concentration in this organ. Phenanthrene was used also to destabilise lysosomal membranes6 in order to test if increased lysosomal fragility interfered with cadmium metabolism and detoxication. The results demonstrated that Cd induced metallothionein synthesis and that elimination of Cd was minimal after 28 days in clear seawater. Lysosomal fragility was initially increased but this effect was soon reversed, even with continued exposure to Cd. The lysosomal destabiliser, phenanthrene, did not appear to affect accumulation of Cd or levels of metallothionein. 相似文献
13.
This laboratory study describes phenanthrene (Ph) and/or anthracene (An) ability to alter the total thiol redox status (TRS), via depletion of protein free thiols (PSH) and glutathione (GSH) levels, in gills of mussel Mytilus galloprovincialis, after a short-term (7 days) exposure period to each contaminant (at a final concentration of 0.1 mg L−1) or in a mixture (ration 1:1, at a final concentration of 0.2 mg L−1). A number of observable changes, like lysosomal membrane impairment (as detected via the neutral red retention time assay, primarily performed in haemocytes), enhancement of lipid peroxidation byproducts, increased nuclear abnormalities, inhibition of AChE and ALP activity, as well as a significant depletion of PSH and GSH were detected in gills of exposed mussels, in any case. Significant relationships occurred among TRS parameters with each change/stress indices measured in tissues of mussels, could reinforce the use of PSH as a potent biomarker. 相似文献
14.
Mike Boryslawskyj Tony Garrood Mike Stanger Terry Pearson David Woodhead 《Marine environmental research》1988,24(1-4)
The aim of this investigation is to understand the factors influencing uptake of persistent organochlorine pesticides into a freshwater mussel (Sphaerium corneum). Previous work has shown that although a substantial proportion of dieldrin is bound to particulates in the river system, uptake is from the dieldrin in solution. This suggests partitioning of the hydrophobic xenobiotics into membranes as the major route of entry. Membrane composition was established using lipid fractionation into neutral lipids and phospholipids followed by derivatisation to fatty acid methyl esters and estimation using GC/FID. Membrane composition varied with acclimation temperature over the range 5–20°C. Dieldrin concentrations in the mussel at equilibrium with 2000 ng dieldrin/litre water burden were studied at one temperature under laboratory conditions using animals acclimated to different temperatures. Any differences may be explicable in terms of effects of membrane composition on partitioning or other physiological adaptations. To establish which of these factors is most significant a membrane fragment partition system has been analysed in vitro. Preliminary results indicate that although water/lipid partitioning may be important in entry (and exit) of non-polar xenobiotics, significant differences could not be found using current methods. The role of membrane composition and therefore temperature history of the organism may still be significant. The implications of this for attempted correlations between bioaccumulation factors and simple octanol/water partition coefficients are discussed. 相似文献
15.
Lysosomal destabilization assays have been used as valuable biomarkers of pollutant exposures in a variety of bivalve and fish species. The responses of oysters, Crassostrea virginica, deployed at and native to various reference and degraded sites were evaluated for lysosomal destabilization during both summer and winter seasons. In both native and deployed oysters, lysosomal destabilization rates tended to be higher during the winter at both reference and polluted sites. There are at least two hypothetical explanations. Greater lysosomal destabilization rates may be related to physiological changes associated with mobilization of nutrient reserves during the winter and gametogenesis. However, lysosomal destabilization in deployed oysters was correlated with tissue metal concentrations. These data also support a second hypothesis that seasonal differences in physico-chemical factors (such as reduced levels of acid volatile sulfides) may increase the bioavailability of metals during the winter so that adverse effects are more pronounced. 相似文献
16.
17.
Polycyclic aromatic pollutants produced by a range of industrial processes and the combustion of fossil fuels. They eventually enter the marine environment, and are bioaccumulated in the tissues of sessile, filter-feeding bivalves, causing a variety of sublethal effects. Few studies, however, have examined immune inhibition in molluscs by these xenobiotics. Here, the immunotoxic effects of PAHs were investigated in marine mussels (Mytilus edulis) by means of in vivo exposure experiments. Animals were exposed to a PAH cocktail of anthracene, fluoranthene and phenanthrene for 2 or 4 weeks. The haemolymph was then extracted, and phagocytosis and lysosomal neutral red retention assays performed to examine the effects of PAHs on particle uptake by haemocytes and lysosome integrity, respectively. PAHs were found to inhibit phagocytosis and damage lysosomes. It is hypothesized that PAHs exert their toxicity directly on the lysosomes. A possible consequence of such lysosomal damage will be immune impairment in mussels leading to reduced resistance to infectious diseases. 相似文献
18.
The purpose of these studies was to evaluate if there were relationships between lysosomal destabilization or glutathione concentrations and gamete viability of oysters, Crassostrea virginica. Oysters were collected from field sites during the peak spawning period (May-June) during 2001 and 2002. Lysosomal destabilization rates and glutathione concentrations of hepatopancreas tissues (e.g. digestive gland) were determined. Eggs and sperm from the same adults were also used to conduct embryo development assays with reference seawater collected from a clean site, site water, and also a range of Cd concentrations (the Cd exposures were used to determine if there were differences in susceptibility to pollutants). Baseline embryo development success (e.g. percent normal development when the assays were conducted with reference seawater or site water) was related to lysosomal destabilization, but not glutathione status. However, the susceptibility of embryos to metal toxicity was related to glutathione status, i.e., sensitivity to Cd exposures increased with decreasing glutathione levels. These studies support the hypotheses that there are linkages between embryo development success and susceptibility to pollutant stress and cellular biomarker responses. These kinds of effects on reproductive success could lead to subtle but significant long-term effects on recruitment and viability of oyster populations. 相似文献
19.
Environmental prognostics: an integrated model supporting lysosomal stress responses as predictive biomarkers of animal health status 总被引：1，自引：0，他引：1
The potential prognostic use of lysosomal reactions to environmental pollutants is explored in relation to predicting animal health in marine mussels, based on diagnostic biomarker data. Cellular lysosomes are already known to accumulate many metals and organic xenobiotics and the lysosomal accumulation of the carcinogenic polycyclic aromatic hydrocarbon 3-methylcholanthrene (3-MC) is demonstrated here in the hepatopancreatic digestive cells and ovarian oocytes of the blue mussel. Lysosomal membrane integrity or stability appears to be a generic indicator of cellular well-being in eukaryotes; and in bivalve molluscs it is correlated with total oxygen and nitrogen radical scavenging capacity (TOSC), protein synthesis, scope for growth and larval viability; and inversely correlated with DNA damage (micronuclei), as well as lysosomal swelling (volume density), lipidosis and lipofuscinosis, which are all characteristic of failed or incomplete autophagy. Integration of multiple biomarker data is achieved using multivariate statistics and then mapped onto "health status space" by using lysosomal membrane stability as a measure of cellular well-being. This is viewed as a crucial step towards the derivation of explanatory frameworks for prediction of pollutant impact on animal health; and has facilitated the development of a conceptual mechanistic model linking lysosomal damage and autophagic dysfunction with injury to cells, tissues and the whole animal. This model has also complemented the creation and use of a cell-based bioenergetic computational model of molluscan hepatopancreatic cells that simulates lysosomal and cellular reactions to pollutants. More speculatively, the use of coupled empirical measurements of biomarker reactions and modelling is proposed as a practical approach to the development of an operational toolbox for predicting the health of the environment. 相似文献
20.
The purpose of these studies was to evaluate if there were relationships between lysosomal destabilization or glutathione concentrations and gamete viability of oysters, Crassostrea virginica. Oysters were collected from field sites during the peak spawning period (May–June) during 2001 and 2002. Lysosomal destabilization rates and glutathione concentrations of hepatopancreas tissues (e.g. digestive gland) were determined. Eggs and sperm from the same adults were also used to conduct embryo development assays with reference seawater collected from a clean site, site water, and also a range of Cd concentrations (the Cd exposures were used to determine if there were differences in susceptibility to pollutants). Baseline embryo development success (e.g. percent normal development when the assays were conducted with reference seawater or site water) was related to lysosomal destabilization, but not glutathione status. However, the susceptibility of embryos to metal toxicity was related to glutathione status, i.e., sensitivity to Cd exposures increased with decreasing glutathione levels. These studies support the hypotheses that there are linkages between embryo development success and susceptibility to pollutant stress and cellular biomarker responses. These kinds of effects on reproductive success could lead to subtle but significant long-term effects on recruitment and viability of oyster populations. 相似文献