Tissue and sub-cellular distribution of Fe,Cu, Zn and 210Po in the abalone Haliotis rubra

The tissue and sub-cellular distribution of Fe, Cu, Zn and the naturally occurring radionuclide polonium-210 was determined in the gastropod mollusc Haliotis rubra collected from Western Port Bay, Australia, between March and July 1988. The highest concentrations of the metals, with the exception of Cu, were found in the digestive gland. Copper was more uniformly distributed, with tissues that are more vasculated having higher concentrations. Ultrastructural examination of the digestive gland, gill and kidney showed dense membrane-bound granules within the cytoplasm. Elemental analysis of the granules by electron probe x-ray microanalysis indicated that the granules in the digestive gland and gill contained high concentrations of iron, with small amounts of copper and zinc. In contrast, the metal-containing granules in the kidney were predominantly composed of iron, copper and phosphorus, with variable contributions of sodium potassium, and calcium. Homogenisation and fractionation of the digestive gland by differential centrifugation confirmed that approximately 80, 10, 90 and 50% of the total homogenate Fe, Cu, Zn and ²¹⁰Po, respectively, sedimented at 1200xg. In the haemolymph, all the elements studied were associated with the soluble high molecular weight component of the serum, not with the amoebocytes. ²¹⁰Po was present in the mucus-secreting hypobranchial glands at about half the concentration found in gill tissue.     

Effects of dietary copper exposure on accumulation,growth, and hematological parameters in Cyprinus carpio

The influence of dietary copper (Cu) exposure on accumulation, growth, and hematological parameters was investigated in Cyprinus carpio after sub-chronic ingestion of 0, 250, 500, 750 or 1000?mg?kg^?1 for 60 days. The profile of Cu accumulation among tissues in C. carpio was dependent on the exposure period and Cu concentration. Liver of C. carpio was the predominant storage tissue and the order of Cu accumulation in tissues was liver?>?intestine?> gill?>?kidney?>?muscle. Cu concentration at >125?mg?kg^?1 reduced growth rate, and was inversely related to growth. The RNA?:?DNA ratios were not affected by exposure and there was no correlation between growth rate and RNA?:?DNA ratio in liver and muscle. There were no significant effects of exposure on blood parameters except for magnesium. Cu exposure time and dose increased the serum glutamic-oxaloacetic transaminase (SGOT) and serum glutamic-pyruvic transaminase (SGPT) activity levels.     

Copper and zinc interaction on water clearance and tissue metal distribution in the freshwater mussel, Cristaria plicata, under laboratory conditions

Copper and zinc interaction on clearance from water and distribution in different tissues was investigated for the freshwater mussel, Cristaria plicata, under laboratory conditions. Clearance rate of Cu or Zn from water was highly dependent on exposure concentration. Interaction effect was most evident at 300 ??g??L^?1 Cu exposure and depressed the Zn clearance rate significantly (p<0.05). However, the presence of 100 ??g??L^?1 and 300 ??g??L^?1 Zn hardly affected the Cu clearance rate. The 300 ??g??L^?1 Cu presence enhanced Cu accumulation in each tissue most significantly (p<0.01), but caused Zn content to decrease in the gills by 62% (p<0.05), viscera by 49% (p<0.05) and foot by 31% (p<0.05), and increase in the mantle by 97% (p<0.05) and the muscles by 243% (p<0.05) for different Zn exposure treatments. The response of metal accumulation in various tissues of the test mussels indicated that Zn transferred from the gills, viscera and foot to the mantle and muscles might be one of the important characteristics of the Zn regulatory mechanism by leading to a narrow range of Zn concentration in the different tissues.     

Effects of environmental hypercapnia on hemato-immunological parameters,carbonic anhydrase,and Na+, K+-ATPase enzyme activities in rainbow trout (Oncorhynchus mykiss) tissues

In this study, the effects of environmental hypercapnia on hemato-immunological parameters and the activities of respiratory enzymes such as carbonic anhydrase (CA) and Na⁺, K⁺-ATPase were investigated in rainbow trout (Oncorhynchus mykiss) tissues (gill, liver and kidney). Batches of 12 fish were exposed to 4.5 mg L^?1 (control) and 14 mg L^?1 CO₂. No mortalities occurred during the 14 days of the experimental period. Red blood cell (RBC), hemoglobin (Hb), and hematocrit (Ht) levels, and innate immune parameters such as nitro blue tetrazolium (NBT), lysozyme, and myeloperoxidase activities, and the melano-macrophage frequency were negatively affected by elevated CO₂ levels. Patterns of change in CA activity differed among the gill, liver, and kidney. Compared with the activities of CA in the control group, the CA enzyme was significantly stimulated at day 7 in the gill tissue, whereas it was stimulated at day 14 of the experiment in the liver tissue of fish exposed to 14 mg L^?1 CO₂ (P < 0.05). In contrast to the pattern of CA enzyme activities, the Na⁺, K⁺-ATPase enzymes were stimulated significantly in the liver after day 7 but inhibited in the kidney and gill (P < 0.05). These results suggest that a subchronic exposure to hypercapnia of rainbow trout tissues may lead to adaptive changes in the respiratory enzymes and negatively affects hemato-immunological parameters.     

Phenotypic variations in the gills of the symbiont-containing bivalve Lucinoma aequizonata

The marine bivalve Lucinoma aequizonata (Lucinidae) maintains a population of sulfide-oxidizing chemoautotrophic bacteria in its gill tissue. These are housed in large numbers intracellularly in specialized host cells, termed bacteriocytes. In a natural population of L. aequizonata, striking variations of the gill colors occur, ranging from yellow to grey, brown and black. The aim of the present study was to investigate how this phenomenon relates to the physiology and numbers of the symbiont population. Our results show that in aquarium-maintained animals, black gills contained fewer numbers of bacteria as well as lower concentrations of sulfur and total protein. Nitrate respiration was stimulated by sulfide (but not by thiosulfate) 33-fold in homogenates of black gills and threefold in yellow gill homogenates. The total rates of sulfide-stimulated nitrate respiration were the same. Oxygen respiration could be measured in animals with yellow gills but not in animals with black gills. The cumulative data suggest that black-gilled clams maintained in the aquarium represent a starvation state. When collected from their natural habitat black gills contain the same number of bacteria as yellow gills. Also, no significant difference in glycogen concentrations of the host tissues was observed. Therefore, starvation is unlikely the cause of black gill color in a natural population. Alternative sources of nutrition to sulfur-based metabolism are discussed. Denaturing gradient gel electrophoresis (DGGE) performed on the different gill tissues, as well as on isolated symbionts, resulted in a single gill symbiont amplification product, the sequence of which is identical to published data. These findings provide molecular evidence that one dominant phylotype is present in the morphologically different gill tissues. Nevertheless, the presence of other phylotypes cannot formally be excluded. The implications of this study are that the gill of L. aequizonata is a highly dynamic organ which lends itself to more detailed studies regarding the molecular and cellular processes underlying nutrient transfer, regulation of bacterial numbers and host–symbiont communication. Received: 1 September 1999 / Accepted: 1 February 2000     

Ontogenic change of gill chloride cells in leptocephalus and glass eel stages of the Japanese eel, <Emphasis Type="Italic">Anguilla japonica</Emphasis>

The development of gill chloride cells was examined in premetamorphic larvae (leptocephali) and juveniles (glass eels) of the Japanese eel, Anguilla japonica. Branchial chloride cells were detected by immunocytochemistry using an antiserum specific for Na⁺,K⁺-ATPase. The specificity and availability of the antiserum for the detection of Japanese eel chloride cells were confirmed by Western blot analysis. The chloride cells first appeared on the developing gill filaments in a mid larval stage of leptocephalus (32.2 mm). Both immunoreactivity and the number of chloride cells gradually increased as the fish grew to a late stage of leptocephalus over 54 mm. In glass eels just after metamorphosis, gill lamellae developed from the gill filaments, and a rich population of chloride cells was observed in the gill filaments. In glass eels collected at a coastal area, chloride cells were extensively distributed in the gill filaments. The chloride cell size decreased progressively in glass eels transferred from seawater (SW) to freshwater (FW), whereas there was no difference in cell number. In contrast, some Na⁺,K⁺-ATPase immunoreaction distinct from typical chloride cells was observed in the gill lamellae throughout FW-transferred fish, but disappeared in control fish maintained in SW for 14 days. These findings indicate that the gill and gill chloride cells developed slowly during the extremely long larval stage, followed by rapid differentiation during a short period of metamorphosis. The excellent euryhalinity of glass eels may be due to the presence of the filament chloride cells and lamellar Na⁺,K⁺-ATPase-immunoreaction, presumably being responsible for SW and FW adaptation, respectively.     

Acid–base regulation in the Dungeness crab (Metacarcinus magister)

Homeostatic regulation allows organisms to secure basic physiological processes in a varying environment. To counteract fluctuations in ambient carbonate system speciation due to elevated seawater pCO₂ (hypercapnia), many aquatic crustaceans excrete/accumulate acid–base equivalents through their gills; however, not much is known about the role of ammonia in this response. The present study investigated the effects of hypercapnia on acid–base and ammonia regulation in the Dungeness crab, Metacarcinus magister on the whole animal and isolated gill levels. Hemolymph pCO₂ and [HCO₃ ^?] increased in M. magister acclimated to elevated pCO₂ (330 Pa), while pH remained stable. Additionally, hemolymph [Na⁺], [Ca²⁺], and [SO₄ ^2?] were significantly increased. When challenged with varying pH during gill perfusion, the pH of the artificial hemolymph remained relatively unchanged. Overall, ammonia production and excretion, as well as oxygen consumption, were reduced in crabs acclimated to elevated pCO₂, demonstrating that either (amino acid) oxidation is reduced in response to this particular stress, or nitrogenous wastes are excreted in an alternative form.     

Pathological effects of sublethal diazinon on the blood,gill, liver and kidney of the freshwater fish Capoeta damascina

Capoeta damascina has a large distribution in the Middle East and is one of the most abundant fishes in inland waters of Iran. Histopathological symptoms of the blood, gill, liver and kidney of C. damascina were investigated over 1, 5 and 9-day exposure to 0 (control), 0.5, 1 and 1.5?mg?L^?1 diazinon. The data were analysed using analysis of variance and non-parametric multivariate analysis of variance. Diazinon concentration was more important than the exposure time on changes of the tissues. Diazinon decreased red blood cell (RBC), white blood cell (WBC), haematocrit (Hct) and haemoglobin (Hb). The symptoms of the gill were mainly desquamation, epithelial hyperplasia, hyperplasia at the base of secondary lamella, epithelial lifting and curling. The prevalent symptoms of the liver were degeneration of nuclei, alternation in size and shape of hepatocytes, focal necrosis and pyknosis. Degeneration of Bowman's capsule, necrosis in renal tubule and haematopoietic tissue were the most common symptoms found in the kidney. In conclusion, the blood, gill, liver and kidney of C. damascina are sensitive enough to respond to low concentrations of diazinon over a short period and this species can thus be a bioindicator of diazinon.     

Histology of Sculpin spp. in East Greenland. II. Histopathology and trace element concentrations

Abstract

For many years, the sculpin has been utilized as a sentinel monitoring species for anthropogenic impacts on the marine environment. To further develop its potential as a screening model body burden of several trace elements, including cadmium (Cd), copper (Cu), mercury (Hg), lead (Pb) and selenium (Se), were investigated. In addition, histopathology in shorthorn sculpins (Myoxocephalus scorpius; n?=?20) and fourhorn sculpins (Myoxocephalus quadricornis; n?=?10) were examined at three sites around the Scoresby Sound settlement in Central East Greenland. Fourhorn sculpins caught at the most distant site from the settlement, contained significantly higher hepatic Cu and Cd concentrations as well as significantly elevated gill Pb levels compared to the shorthorn sculpins collected at two other sites closer to the settlement. Histological examinations showed that fish with significantly higher concentrations of hepatic Cd and Se exhibited greater nuclear alterations, interstitial hyperplasia/hypertrophy, interstitial mononuclear infiltrations and granulomas. Further, fish with higher Cd and Cu gill tissue levels displayed a significantly higher number of cytoplasmic alterations and lamellar epithelium lifting, hypertrophic and hyperplastic epithelium along with mucus cell hyperplasia. While the presence of liver lesions were not species or sex-specific, the presence of gill lesions decreased in the order female fourhorn sculpins?>?female shorthorn sculpins?>?male fourhorn sculpins?>?male shorthorn sculpins. Hepatic Hg concentrations exceeded known lowest observable effect doses (LOED) for fish (0.1–0.5?μg g^?1 ww) in 27% of fish, while liver Cd residues in 80% exceeded LOED (0.42–1.8?μg g^?1 ww). Based upon these results, data suggest that using the sculpin as a valuable sentinel fish species histopathology may serve as a reliable tool for assessing marine ecosystem exposure to trace metals. However, confounding physiological and ecological factors also need to be considered.     

单独与混合暴露条件下褶纹冠蚌对Cu、Zn的积累与分布

采用室内模拟方法,在单独与混合暴露条件下研究了褶纹冠蚌(Cristaria plicata)对Cu、Zn的积累与分布特征.结果表明:褶纹冠蚌对水环境中的Cu、Zn具有良好的积累能力,持续暴露8d内,体内各组织重金属积累量基本呈线性上升,其中鳃与外套积累速率最快(p<0.05).Cu在褶纹冠蚌鳃、内脏团和生殖腺中的积累速率与Zn之间存在显著差异(p<0.05),且Cu、Zn之间在各组织中的积累拮抗作用明显.暴露前,Cu含量在褶纹冠蚌体内依次为:生殖腺>外套、内脏团>鳃、足>肌肉(p<0.05),变化幅度为10倍左右;Zn含量依次为:鳃>内脏团>生殖腺>足、外套>肌肉(p<0.05),变化幅度为4倍左右.暴露后,外套和鳃对Cu、Zn的积累量最高,且Cu、Zn含量在不同组织间的变化幅度明显增加.暴露前后,Cu含量在不同组织间的变化幅度均高于Zn.不同暴露条件下,褶纹冠蚌不同组织对Zn的浓缩系数(BCF)明显高于Cu.     

Lead- and cadmium-induced histopathological changes in gill,kidney and liver tissue of freshwater climbing perch Anabas testudineus (Bloch, 1792)

The objective of this study was to examine the toxicological effect of two major heavy metal pollutants, lead chloride (PbCl₂) and cadmium chloride (CdCl₂), in the freshwater climbing perch, Anabas testudineus. Fish were exposed to 1.0 and 2.0 mg/L both metals and histopathological changes in gill, kidney and liver tissues were studied. Major changes observed in gill tissue were epithelial lifting, proliferation of epithelial cells, fusion of secondary lamellae, hyperplasia and hypertrophy of mucous cells, and necrosis of epithelial cells. Cell necrosis, degenerated kidney tubules, congestion, lymphocytic infiltration and vacuolation were the major abnormalities observed in kidney tissue. The most conspicuous changes in liver were darker nucleoli, irregularly shaped hepatocytes with dilated blood capillaries, and focal as well as single necrosis. Fish specimens exposed to PbCl₂ exhibited pronounced changes in all tissues examined compared with those exposed to CdCl₂. It is evident from this study that heavy metals can cause significant histopathological changes in fish tissue.     

Effects of UV-B radiation on the gills of Catla catla during early development

Scanning electron microscopic (SEM) study of gills of Catla catla catla (17-day-old) exposed to UV-B radiation (145?µW?cm^?2 at the water surface) for three different exposure times: 5, 10 and 15?min was conducted. Fish without UV-B exposure served as control. UV-B radiation damaged both gill filaments and lamellae. The intensity of damage was minimal in 5?min exposed fish, followed by 10?min exposed fish and maximal in 15?min. The gill epithelium was severely damaged in 15?min irradiated fish compared to control. Pavement cells (PVCs) were damaged and the numbers of microridges within PVCs decreased. The deep boundary of PVC was not clear. In some area of gill epithelium, PVCs were destroyed and mitochondrion-rich cells (MRCs) were exposed. The 5?min exposure reduced the number of microridges in the PVCs, but the boundary of PVCs was still visible. MRCs in the gill epithelium were not exposed in 5?min exposed fish. The damage to PVCs and subsequent exposure of MRCs in UV-B irradiated fish may hamper respiratory functions and disturb osmoregulation in catla.     

铜离子对中国花鲈幼鱼的毒性研究

为给中国花鲈(Lateolabrax maculatus)养殖提供污染生物学的理论数据,研究了中国花鲈幼鱼的铜离子中毒症状和半致死浓度(LC50);检测了幼鱼肝脏的谷胱甘肽(GSH)和丙二醛(MDA)含量及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷丙转氨酶(GPT)和谷草转氨酶(GOT)活力;分析了铜离子胁迫对...     

铜在日本青鳉体内的累积特征及其对肝脏组织结构的影响

为了解水体铜(Cu(Ⅱ))在日本青鳉(Oryzias latipes)脑、鳃、肝脏、性腺和肌肉组织中的累积特征,以及Cu(Ⅱ)对肝脏组织结构的影响,本文采用生态学单因子梯度实验,进行20 d的日本青鳉毒性暴露试验。结果显示,日本青鳉各个组织对Cu(Ⅱ)的累积量随着浓度的升高而增多,但不同组织对Cu(Ⅱ)的累积量有较大的差异,铜累积量由大到小依次为:肝脏脑鳃性腺肌肉。铜暴露后的日本青鳉肝细胞出现肿大、大小不一、排列混乱,静脉扩张淤血,部分肝细胞固缩和坏疽等现象。肝脏是日本青鳉蓄积铜的主要器官之一,实验发现高浓度的Cu(Ⅱ)对肝脏组织结构造成了明显的损伤,进而严重影响日本青鳉的存活率。     

Studies on the fusion reaction in two species of Perophora (Ascidiacea)

Histological and electron microscopical studies were made on the fusion reaction between homogeneic stolons of the colonial ascidians Perophora japonica Oka and P. sagamiensis Tokioka. When two stolons make a tip-to-side contact, the epidermal cells of the lateral portion of one stolon become thick and form a protrusion. In the fusion reaction of P. japonica, epidermal cells of each stolon at the contact area extend cell processes immediately after contact. After an occurrence of degenerated epidermal cells, blood interchange between the two stolons is initiated by a partial elimination of the epidermal cells. Even after the establishment of blood interchange, the cuticlelike structure is often observed to remain at the contact area. In the fusion reaction of P. japonica, many amoebocytes are attached to the epidermal cells until the complete elimination of epidermal cells. In the fusion reaction of P. sagamiensis, many lymphocytes as well as amoebocytes are attached to the epidermal cells at the contact area.Contribution No. 461 of the Shimoda Marine Research Center     

Effects of three different nanoparticles on bioaccumulation,oxidative stress,osmoregulatory, and immune responses of Carcinus aestuarii

Abstract

In this study, the toxicity of CuO (40?nm), α-Al₂O₃ (40?nm), and α-Fe₂O₃ (20–40?nm) nanoparticles was comparatively investigated on Carcinus aestuarii. Crabs were semi-statically exposed to 1?mg/L of each for 14?days and their accumulation and distribution in tissue and hemolymph, potential oxidative stress mechanism, total hemocyte counts and types, and the osmoregulatory and ionoregulatory responses were determined. The tissue distribution of CuO nanoparticles was hepatopancreas?>?hemolymph?≥?gill?> muscle, for α-Fe₂O₃ gill?>?hepatopancreas?>?muscle?> hemolymph, and for α-Al₂O₃ gill?>?muscle?≥?hemolymph?> hepatopancreas. While α-Al₂O₃ and α-Fe₂O₃ NPs, induced lipid peroxidation and changes in antioxidant enzyme activity in the hepatopancreas tissue, the oxidative damage caused by the CuO nanoparticles was minimal. All three nanoparticles, copper in particular, elicit osmoregulatory and ionoregulatory toxicity at this concentration, due to the inhibition of Na⁺, K⁺-ATPase activity in the gill and depletion of hemolymph and carcass ion concentrations.     

Ultrastructure of the gill of the hydrothermal-vent mytilid Bathymodiolus sp.

Specimens of Bathymodiolus sp. were collected at 2 620 m depth during the Biocyarise Cruise (12°5880N; 13°5660W) in March 1984, and samples of the gill were fixed for ultrastructural observations. The gill of this hydrothermal-vent mytilid constitutes the main organ in the nutritional processes. The lamellae display abundant ciliation, normally comprised of frontal cilia, compound latero-frontal cirri and lateral cilia. At the ventral margin of each demi-branch, a longitudinal, ciliated, feeding groove is present. the lamellae are composed of numerous homorhabdic filaments connected by tufts of cilia. Each filament is made of a thin wall overlying a central lumen containing amoebocytes. Ultrastructural observations revealed the filament wall to be composed of four types of cells: (1) The ciliated cells of the frontal, latero-frontal and lateral ciliation, characterized by an abundance of mitochondria. (2) Mucous cells present to some degree among the ciliated cells, but more abundant on the distal edge and containing dense droplets of mucus. (3) Cells colonized at their apical pole by numerous bacteria enclosed in membrane-delimited clear spaces and composing the major part of the filament wall. (4) Thin ciliated cells separating the bacterial cells and characterised by a dense fringe of microvilli at their apical pole. The lumen of the filament contains amoebocytes of different morphological aspects which seem to accumulate electron-dense granules, possibly related to detoxification processes.     

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.     

Histopathological effects of copper oxide nanoparticles on the gill and intestine of common carp (Cyprinus carpio) in the presence of titanium dioxide nanoparticles

In current research, the combined effects of copper oxide nanoparticles (CuO NPs) and titanium dioxide nanoparticles (TiO₂ NPs) on the histopathological anomalies of gill and intestine tissues in common carp (Cyprinus carpio) were studied. Common carp were exposed to TiO₂ NPs (10.0?mg L^?1), CuO NPs (2.5 and 5.0?mg L^?1), and mixture of TiO₂ NPs (10.0?mg L^?1)?+?CuO NPs (2.5 and 5.0 mg?L^?1) for two periods of exposure (10 and 20 days) and recovery (30 and 40 days). The most common histopathological anomalies in the gill of common carp such as hyperplasia, oedema, curvature, fusion, aneurism, and necrosis were observed. The synergistic effect of co-existing TiO₂ NPs and CuO NPs reduced the length of secondary lamella and increased the diameters of the gill filaments and secondary lamellae. Moreover, the presence of TiO₂ NPs increased the CuO NPs effects on the histopathological anomalies of intestine tissue and the synergistic effect of TiO₂ NPs and CuO mixture leads to an increase in the severity of histopathological lesions such as degeneration, swelling of goblet cells, and necrosis - erosion in the intestine tissue. In conclusion, the presence of TiO₂ NPs increased the toxicity of CuO NPs.     

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.