Histochemical demonstration of mercury in the intestine,nephridia and epidermis of the marine polychaete Nereis virens exposed to inorganic mercury

Marine polychaetes Nereis virens (Sars) were exposed to 9 ppb Hg as mercuric chloride in the aquarium water. Concentrations of mercury in the water and in the worms were monitored using radiolabelled mercury. After 11 d, the average mercury concentration in the nereids was 8.41 ppm Hg and the bioconcentration factor was calculated to be 930. Accumulations of mercury in the tissues were made visible for light and electron microscopy by autometallographic silver enhancement. Mercury was demonstrated in the intestine, nephridia, epidermis and cuticula. In the intestine, mercury was found to be located predominantly in the apical part of the epithelial cells. In the nephridia, mercury deposits were apparent in the peritoneal cells of the nephridial tubules. Silver-enhanced mercury was also observed at the epicuticula, in the collagen fibres of the endocuticula and in the epidermal basal cells. Intracellularly, the observed mercury accumulations were localised predominantly in lysosomes. Extracellularly, mercury was observed in the basal laminae of the intestine and the epidermis, and also in the intestinal peritrophic membrane. How N. virens copes with mercury toxicity is discussed.     

Biochemical composition of the blue crab Callinectes sapidus exposed to the water-soluble fraction of crude oil

The biochemical composition of juvenile blue crabs, Callinectes sapidus, exposed to sublethal concentrations of the water-soluble fraction of South Louisiana crude oil (0 to 2 504 ppb) for 21 d were examined. Although growth took place in all crabs, tissue content varied inversely with exposure concentration while percentage tissue water varied directly with exposure concentration. Total protein, lipid and RNA content of crabs exposed to crude oil were significantly less than that of control crabs by Day 21. DNA content was not significantly different from that of control crabs, suggesting that the difference in tissue content was due to differences in cell volume and not cell number. There were no consistent differences in the concentration of the major biochemical components, indicating that the relative contribution of each of the components remained stable during the period of sublethal stress. Ratios of RNA:DNA and protein:DNA decreased in exposed crabs and were positively correlated with scope for growth and observed growth. The ratios may be useful as indirect indicators of physiological condition. Analysis of lipid classes indicated that structural lipids in stressed crabs were less affected than were lipids used for energy storage. The changes in biochemical composition suggest that the pattern of energy utilization was altered in crabs exposed to crude oil. Growth in size without comparable growth in tissue resulted in decreased tissue content. Available energy was used for growth, with little being stored in lipid reserves.     

Sodium regulation in the shore crab Carcinus maenas as related to ambient salinity

The activity of Na–K-ATPase was measured in crude homogenates prepared from various organs (leg muscle, pincer muscle, heart, testes, digestive gland, hypodermis, gills 1–9) of shore crabs, Carcinus maenas L., acclimated to salinities ranging between 10 and 50 S (in steps of 10 S). In all salinities tested, Na–K-ATPase activity was highest in posterior gills 7–9 (10–12 mol P_i mg protein^-1 h^-1), followed by anterior gills 1–6 (ca. 2.5 mol P_i mg protein^-1 h^-1) and the other organs (in most cases far below 2mol P_i mg protein^-1 h^-1). In gills only, Na–K-ATPase activity was salinity-dependent, with the highest values in the lowest salinities and vice versa. In gills 7–9, Na–K-ATPase activity was increased more than threefold following a reduction in salinity from 50 to 10 S. Na–K-ATPase activity, expressed as percentage of total ATPase activity, amounted to 60–80% in gills, about 60% in hypodermis and 20–40% in the other organs. Ouabain, a specific inhibitor of Na–K-ATPase activity, reduced serum osmolalities in crabs kept at 9–10 S only when injected into the hemolymph (1 and 5 · 10^-5 M), but had no effect when dissolved in ambient water (10^-4 M). The results obtained underline that crustacean gills are the main organs for ionic regulation, and confirm the hypothesis of the central role of the Na–K-ATPase in active Na uptake as the basic mechanism of hyperregulation in dilute media. Reduction of serum osmolalities following injection of ouabain into the hemolymph confirms previous reports on localization of the sodium pump in the basolateral parts of epithelial cells.     

Study on the gill of field crab,Paratelphusa hydrodromus (Herbst.) exposed to nickel

Paratelphusa hydrodromus (Herbst.) was exposed to 50 ppm nickel chloride solution for a period of 10 days under laboratory conditions. The gills of the treated and non-treated animals were sectioned, stained and examined under a compound microscope and noticed significant changes such as enlargement of gill lamellae, lifting up and rupture of epithelial cells, enlargement of mid rachis, hyperplasia and hypertrophy, appearance of pyknotic nuclei and a general necrosis in the treated gills. Results suggest that a comparatively low concentration of nickel (50 ppm) is enough to elicit pathological changes in Paratelphusa hydrodromus.     

Osmotic and ionic regulation in the fiddler crab Uca rapax acclimated to dilute and hypersaline seawater

Adult male Uca rapax, collected from the central coast of Venezuela in early 1994 were gradually acclimated to salinities ranging from 1.7 to 139S. The hemolymph osmolality (791±15 mOsmol in crabs from 26S) changed less than three-fold over the entire range of concentrations tested. The urine was isosmotic with the hemolymph in crabs exposed to dilutions <26S, and significantly hyperosmotic in those exposed to media >34.8S. The hemolymph levels of Na⁺, Cl^–, K⁺, Ca²⁺ and Mg²⁺ (320±13, 405±17, 7.8±0.7, 7.2±0.1 and 31±2.2 mmol l^–1, respectively, in crabs acclimated to 26S) were maintained fairly constant over the range from 8.7 to 99S, decreasing by 15% in the more dilute media or increasing sharply to about twice those values in crabs from 139S. The excretory organs contributed to the osmoionic regulation of the hemolymph in crabs exposed to <3.5 or to >34.8S, by means of the partial reabsorption or excretion, respectively, of salts from or into the urine. The results described place U. rapax among the most powerful hypo/hyper-regulating crustaceans known.     

Effects of size upon salinity tolerance and volume regulation in the hermit crab Pagurus bernhardus

Whereas small Pagurus bernhardus (L.) occur in large numbers in the intertidal areas of rocky shores, large hermit crabs living in whelk shells are most uncommon, although they are easily collected by sublittoral dredging. Experiments were carried out to assess the salinity tolerance, and the ability to regulate body volume in a dilute medium, of both small and large hermit crabs. Large, offshore hermit crabs are significantly less tolerant of 60% sea water (100%=34 S) than small, littoral specimens. Small littoral crabs increase in weight by about 15% in the first hour in 60% sea water, but increased urine output prevents swelling thereafter; indeed, there is considerable loss in weight, presumably caused by solute loss. Large, offshore, crabs increase in weight by about 15% in the first hour, thus exhibiting a higher permeability to water than small crabs, and continue to increase in weight, since urine output is not sufficient to cope with the water load. Dissections suggest that insufficient release of urine causes the low urine output in large hermit crabs, rather than inadequate production of primary urine. Calculations indicate that the nephropores of large hermit crabs are too small to allow a sufficiently high urine output to remove a large water load, unless greater pressures are developed inside the excretory systems of large hermit crabs than occur in those of small littoral individuals. In view of the soft, flexible nature of the walls of the excretory system and abdomen of P. bernhardus, it seems unlikely that high pressures could be maintained within the excretory system. It is concluded that the stenohalinity of large, offshore hermit crabs may exclude them from the littoral zone, or may be the result of exclusion by other factors, since there will be no selective advantage for large hermit crabs in maintaining the degree of euryhalinity possessed by the small littoral crabs, if the large crabs are confined to the sublittoral zone.     

酸沉降影响下重庆马尾松林的健康监测及调控技术研究

酸沉降影响下重庆马尾松(Pinus massoniana)林的衰退一直受到研究者的重视,并取得一定成果,但森林健康监测和调控技术等方面的研究还很少。以重庆铁山坪马尾松纯林为研究对象,采用欧洲叶量损失率的划分标准,选择叶量损失率为0～20%、20%～40%、40%～60%和60%～99%的优势木,分析其树冠下的土壤化学性质及针叶元素含量;采用随机区组设计,设立在土壤表面撒施石灰石粉0(不撒施石灰石粉,对照)、1、2、3和4t·hm-25个处理,探讨不同石灰石粉剂量处理对优势木针叶元素含量和生长的影响。结果表明:不同叶量损失率的马尾松优势木树冠下的土壤化学性质基本相似,0～60cm土层的pH平均值在3.59～3.80,盐基饱和度平均值低于10%,土壤严重酸化,随叶量损失率的增加,一年生针叶内K、Na、Ca、Mg、Al、P、S、N等元素含量并无明显差异。针叶内P缺乏是营养限制因子。撒施石灰石粉165d后,1、2、3t·hm-2处理的优势木一年生针叶中P、Mg含量增加,增幅以2t·hm-2处理最高,比对照分别增加30.26%和12.65%;所有撒施石灰石粉处理的N含量增加,Ca含量有增有减,变化规律不明显。撒施石灰石粉一年后,各处理优势木的树高和胸径生长量增加,针叶损失率下降,主轴顶梢长度和一年生针叶长度增加。短期内从一年生针叶中P、Mg含量及林木生长和健康情况变化综合来看,石灰石粉一次撒施量以2t·hm-2效果最佳。     

Influence of chloride ions on the reduction of mercury species in the presence of dissolved organic matter

Environmental Geochemistry and Health - Mercuric species, Hg(II), interacts strongly with dissolved organic matter (DOM) through the oxidation, reduction, and complexation that affect the fate,...     

Gene expression profiles of the swimming crab <Emphasis Type=Italic>Portunus trituberculatus</Emphasis> exposed to salinity stress

The swimming crab, Portunus trituberculatus, is an important marine fishery and aquaculture species. Although P. trituberculatus is a euryhaline species, water salinity condition influenced its distribution, migration route, and artificial propagations. To investigate gene expression in the P. trituberculatus exposed to different salinity stresses, 2426 expressed sequence tags (ESTs) from gill cDNA library were selected to spot on a cDNA microarray chip. In total, 417 differentially expressed genes were identified and grouped into eight clusters by hierarchical clustering analysis. Approximately 71.5% of grouped genes belonged to three independent expression patterns, indicating that these three expression patterns may represent three important stress tolerance pathways or networks in P. trituberculatus. Moreover, our cDNA microarray data suggested that there were differences in gene expression patterns of P. trituberculatus for low salinity and high salinity acclimation, suggesting that two salinity challenges resulted in a wide variation of gene expression in P. trituberculatus. In addition, a series of genes such as CCAAT/enhancer-binding protein, Na/K ATPase β-subunit, and heat shock proteins (HSPs) genes were suggested to be key elements during salinity acclimation process. Overall, this work represented an important step toward understanding the molecular processes and mechanisms involved in salinity acclimation of the swimming crab.     

Effects of sulfide exposure history and hemolymph thiosulfate on oxygen-consumption rates and regulation in the hydrothermal vent crab Bythograea thermydron

The hydrothermal vent crab Bythograea thermydron is exposed to high environmental concentrations of sulfide and low levels of oxygen for extended periods of time. It has previously been shown that hydrogen sulfide is oxidized to the relatively non-toxic thiosulfate (S₂O ₃ ^2– ), which accumulates in the hemolymph. Hemolymph thiosulfate levels in freshly captured crabs vary significantly among crabs from different hydrothermal vent sites as well as between crabs from different microhabitats within the same site. Hemolymph thiosulfate concentrations were not significantly different between crabs captured at the same site 6 mo apart. Hemolymph thiosulfate concentrations ranged from 66 mol 1^–1 in a crab captured at a site with relatively low sulfide venting, to 3206 mol 1^–1 in an individual that was netted from an active smoker vent with much higher sulfide exposure. The differences in hemolymph thiosulfate between sites and the stability of hemolymph thiosulfate in crabs captured at the same site at different times suggest that sulfide exposure is significantly different between sites and that this exposure may not vary significantly over the course of a few months. B. thermydron experimentally exposed to sulfide had high levels of thiosulfate in their hemolymph and increased abilities to regulate oxygen consumption in conditions of low oxygen. This enhancement of regulatory abilities suggests that the previously demonstrated increased hemocyaninoxygen (Hc–O₂) affinity due to elevated thiosulfate may be adaptive in vivo. Average oxygen-consumption rates were much higher in crabs experimentally exposed to sulfide than in unexposed crabs. Crabs injected with isosmotic thiosulfate did not have increased oxygen-consumption rates as did the sulfide-exposed individuals, but did show a similar reduction in P _c (the critical partial pressure of oxygen at which crabs can no longer regulate oxygen consumption). This suggests that it is the sulfide exposure and/or detoxification rather than the elimination of thiosulfate that causes the increase in metabolic rate. Thiosulfate diffuses into dead crabs and into live crabs exposed to 15 mmol S₂O ₃ ^2- l^–1, indicating substantial permeability, and yet live crabs are able to eliminate thiosulfate when incubated in sea water containing 1.5 mmol S₂O ₃ ^2- l^–1, suggesting a process that has an active component.     

Toxicity of the insecticide methoxychlor to the Dungeness crab Cancer magister

The toxicity of methoxychlor, determined in acute and chronic studies using larval, juvenile, and adult stages of the Dungeness crab, Cancer magister Dana, was inversely related to the age of the crabs after hatching and increased with the length of exposure. The 96-h LC₅₀'s for zoeae, juveniles, and adults were 0.42, 5.10, and 130 g/l, respectively. Levels causing decreased survival of these same stages during exposures of more than 60 days were 0.05, 0.40, and 4.0 g/l, respectively. Hatching of eggs was not adversely affected by methoxychlor concentrations up to 10 g/l, and molting from prezoeae to zoeae was reduced only 30% from controls at the latter exposure concentration. Methoxychlor concentrations of 0.05 and 4.0 g/l delayed motting of larval and juvenile crabs, respectively, and this delay was as much as 10 days for the juveniles. Sensitivity of adult and juvenile crabs to methoxychlor increased during ecdysis. In uptake experiments, juvenile crabs concentrated methoxychlor more rapidly than did adults. Juveniles exposed to 2.0 g/l of methoxychlor and adults exposed to 1.8 and 7.5 g/l had whole body methoxychlor concentrations after 12 days of 0.88, 0.10, and 0.51 mg/kg, respectively. Loss of the pesticide from adult crabs was rapid, and was 95% complete after 15 days of depuration. Concentrations of methoxychlor in individual tissues were found to be highest in the exoskeleton, gill, and hepatopancreas, in declining order, with less than a two-fold difference among these tissues. About 81% of the methoxychlor measured in whole body samples was associated with the exoskeleton, but it is not known whether or not the pesticide was transported through the culticle to internal tissues.Technical Paper No. 4133, Oregon Agricultural Experiment Station.     

Metabolic disposition of lactate in the horseshoe crab Limulus polyphemus and the stone crab Menippe mercenaria

The metabolism of ¹⁴C-labelled lactate was investigated in the horseshoe crab Limulus polyphemus and the stone crab Menippe mercenaria. When a bolus of (¹⁴C-U)-D-lactate was injected into L. polyphemus, there was substantial release pf ¹⁴CO₂ into the medium. In the case of M. mercenaria, ¹⁴CO₂ release was also observed after injection of (¹⁴C-U)-L-lactate into experimental individuals. Analysis of the distribution of radioactivity in whole body extracts of both species revealed easily detectable amounts of radioactivity in the glycogen fraction, although the bulk of the radioactivity was in the cation, anion/neutral and CO₂ fractions. To investigate the metabolism of lactate further, ¹⁴C-labelled lactate was injected into large individuals of L. polyphemus and M. mercenaria, and the distribution of radioactivity was determined in the hemolymph, muscle and hepatopancreas. Utilization of (¹⁴C-U)-D-lactate by L. polyphemus resulted in the accumulation of significant amounts of labelled glucose in all three body compartments as well as the production of labelled glycogen in the telson levator muscle and hepatopancreas. Utilization of (¹⁴C-U)-L-lactate resulted in a similar pattern of glucose and glycogen labelling in the hemolymph, cheliped muscle and hepatopancreas of M. mercenaria. These studies demonstrate that both L. polyphemus and M. mercenaria have the capacity for glyco- and gluconeogenesis using lactate as the substrate.     

Osmotic and chloride regulation in the hemolymph of the tiger prawn Penaeus monodon during molting in various salinities

The effect of molting on osmotic and chloride concentrations in the blood of the prawn Penaeus monodon Fabricius (20±3 g) at various salinities was investigated. Prawns were obtained from ponds in Iloilo, Philippines, in 1984. They were stocked in salinities of 8, 20, 32 and 44, and their hemolymph was sampled during molt (Time 0), and then 0.125, 0.25, 0.5, 1, 2, 4, 6, 10 and 14 d after molting. Prawns during and immediately after molt tended to conform to the environmental osmolality. Subsequent postmolt (0.5 d) stages displayed more divergence from external salinity. The isosmotic point was higher (940±30 mOsm kg^-1) during molt than during intermolt (663±8 mOsm/kg^-1), suggesting different osmotic requirements in early molt. Hyperregulation of hemolymph chloride below 20 S, as well as isoionic point (301±6 mM), were independent of molting stage. At 20 S and above, newly molted (0 to 0.25 d post-molt) individuals tended to conform to the external chloride concentration while intermolt (0.5 d) post-molt individuals did not. Contribution of hemolymph chloride to hemolymph osmolality was greater during intermolt than during ecdysis, suggesting an important role for other negatively charged ions during molt. When molt occurred in 20 S (the test salinity most similar to the isoionic salinity), there was little or no change in hemolymph osmolality or chloride concentration from 0 to 14 d postmolt. At 8, 32 and 44 S, the change from molt to intermolt values in hemolymph osmotic and chloride concentrations was hyperbolic. Non-linear least-squares regression showed that prawns generally achieved intermolt values within 1 d after molting. Prawns at intermolt regulated hemolymph osmolality (620 to 820 mOsm kg^-1) and chloride concentration (300 to 450 mM) at a much narrower range than during molt (520 to 1 170 mOsm kg^-1 and 250 to 520 mM, respectively). Hemolymph osmolality was a more sensitive indicator of physiological response than hemolymph chloride concentration. Distribution and culture of P. monodon might be limited in low salinities by its ability to maintain a hemolymph osmolality 500 mOsm kg^-1 during molt and 600 mOsm kg^-1 in intermolt, and in high salinities by its capacity to reduce the hemolymph osmolality from values at molt to those in intermolt. Osmotic and chloride concentrations in the blood of P. monodon clearly varied with both molt stage and salinity of the medium. Dependence on external factors, however, gradually declined in older molt stages, suggesting a reduction in integument permeability and greater development of ion absorption/secretion mechanisms as the exoskeleton hardened.SEAFDEC Contribution No. 197     

Hg2+胁迫下玉米生理生态变化的研究

通过砂培实验,研究汞胁迫下玉米的生理生态指标包括叶绿素含量、根系活力、游离脯氨酸含量的变化.结果表明,汞对玉米的胁迫是浓度和时间双重因子的作用在低汞浓度、短时间内,叶绿素含量、根系活力及游离脯氨酸含量均有上升的趋势;在高汞浓度、长时间内,各生理指标均严重下降.各生理指标对汞的敏感度略有差异游离脯氨酸含量>根系活力>叶绿素含量;其中脯氨酸含量可以作为汞污染的一个监测指标.     

Effect of fuel oil and dispersant on cell cycle and macromolecular synthesis in the chlorococcal alga Scenedesmus armatus

Growth and reproductive processes in synchronous cultures of the alga Scenedesmus armatus (isolated from Baltic phytoplankton) were followed in the presence of various concentrations of dispersant DP-105, oil, and mixtures of oil and dispersant. The inhibition of protoplast fission was the most prominent effect of oil. Nuclear division was inhibited to a lesser extent. With an increasing concentration of oil, dispersant, or a mixture of both, the inhibitory effects were expressed earlier. The presence of oil compounds did not effect the timing of DNA replication, but it did reduce the number of replication rounds in a concentration dependent manner. The inhibition of DNA synthesis was accompanied by slightly delayed cessation of RNA and protein synthesis. Starch synthesis was always inhibited to a lesser extent, and at a later time, than other macromolecular syntheses. Pigment synthesis continued almost to the end of the cell cycle. Thereafter, a rapid degradation of all pigments began and the cells became bleached. No inhibitory effect on reproductive processes was found if oil or dispersant were added to cultures transferred into darkness. Furthermore, recovery from inhibition of reproductive processes caused by oil in continuously illuminated cultures was observed in darkened cells. Chemically dispersed fuel oil was only slightly more toxic than mechanically dispersed fuel oil; an additive, rather than synergistic, effect of oil/dispersant mixture was found.     

Rhythmic activity responses of the fiddler crab Uca crenulata to artificial tides and artificial light

The fiddler crab Uca crenulata, from California (USA), was exposed to artificial tides in order to differentiate between the influence of a tidal cycle from that of a light-dark cycle on its locomotor activity. Most crabs could be well synchronized by tides, but the activity patterns of other crabs was merely exogenously reinforced. Under constant conditions, after tidal treatment, crabs showed bimodal or unimodal activity patterns. When a 24 h light-dark cycle and a 12.35 h tidal cycle acted as concurrent stimuli, their effectiveness in synchronizing the crabs' activity was variable, depending on the undividual. It is likely that crabs which tend to exhibit a unimodal activity pattern are more sensitive to a light stimulus, whereas crabs with a bimodal activity pattern respond preferably to the tides. Thus, responses of endogenous activity of U. crenulata to tides and light-dark cycles are similar, suggesting that the basic oscillarory mechanism for circadian and tidal activity may be the same.     

Rhythms in larval release of the sublittoral crab Neopanope sayi and the supralittoral crab Sesarma cinereum (Decapoda: Brachyura)

Ovigerous females of the subtidal xanthid crab Neopanope sayi (Smith) and the high intertidal grapsid crab Sesarma cinereum (Bosc) were collected during the summers of 1986 and 1987 in the Beaufort, North Carolina (USA), area and brought into the laboratory, where rhythms in larval release were monitored. When crabs with late-stage embryos were put under a 14 h light:10 h dark cycle in an otherwise constant-environment room, an apparent tidal rhythm in release of larvae was observed for both species, with N. sayi releasing near the time of day and night high tides, and S. cinereum releasing around the time of night high tides. The time of sunset relative to high tide was a complicating factor, since larval release for both species was often concentrated around sunset when evening high tides fell several hours before sunset. When a group of N. sayi and S. cinereum were brought into the laboratory and placed under constant lowlevel light for 5 d, the release rhythm of the population persisted, thus implying that the rhythm is endogenous. Larval release near the time of high tide and often at night is common among brachyurans living in tidal areas, regardless of specific adult habitat, suggesting a common functional advantage. Possibilities include transport of larvae from areas where predation and the likelihood of stranding and exposure to low-salinity waters are high, as well as a reduced probability of predation on adult females. Results of the present study suggest that the importance of release after darkness may increase with increasing tidal height of the adult.     

Environmental levels of sediment-associated tri-n-butyltin chloride (TBTCl) and ionic regulation in flounders during seawater adaptation

The effects of exposure to sediment-associated tri-n-butyltin chloride (TBTCl) were examined in the euryhaline European flounder, Platichthys flesus (L.). The effects were quantified by measuring the changes in sodium efflux; Na⁺/K⁺-ATPase activity; and the numbers, areas, and distribution of chloride cells in the gills of freshwater-adapted fish, following a rapid transfer to seawater. Following the transfer to seawater, the Na⁺/K⁺-ATPase activity and the sodium efflux were significantly increased in the control group but remained unchanged in the TBTCl-exposed group. The normal morphological changes to the gill epithelium associated with seawater adaptation, which involve chloride cell distribution, took place in the control group but were significantly inhibited or delayed in the TBTCl group. The results presented in this study lead to the conclusion that environmental concentrations of tri-n-butyltin chloride in sediments are capable of significantly disrupting both the physiological and the morphological components of iono-regulatory functions of an estuarine flatfish.     

Use of mercury in agriculture and its relationship to environmental pollution

Linuron samples at 75mg/L concentration were photolyzed with ultraviolet lamps or with natural sunlight. Ten photoproducts were identified and yields were determined for the more significant photoproducts resulting from either sunlight or ultraviolet lamp photolysis.