鱼类甲状腺轴对胚胎发育、仔鱼变态及性别分化的调控作用研究进展

鱼类甲状腺轴由下丘脑、垂体、甲状腺组成,下丘脑通过分泌促甲状腺激素释放激素、促肾上腺皮质激素释放激素调节垂体促甲状腺激素的合成和分泌,促甲状腺激素促进甲状腺滤泡合成并释放甲状腺激素(主要为甲状腺素—T4),T4运输至肝脏等外周组织中,经过脱碘转化形成生物活性更强的三碘甲腺原氨酸(T3),T3主要通过同核受体结合发挥生物学功能。甲状腺激素对鱼类胚胎发育、仔鱼变态及性别分化等早期生长发育过程具有重要调控作用:甲状腺激素为鱼类胚胎发育期所必须,外加适量甲状腺激素能够提高胚胎或仔鱼存活率;仔鱼变态过程中,某些外部形态变化及相关组织分化依赖甲状腺激素的调控,牙鲆等仔鱼变态期体内甲状腺激素水平及其受体表达量会显著升高;性腺发育期,甲状腺激素对调控鱼类性别分化方向可能具有一定作用。     

方斑东风螺早期发育过程中几种消化酶的活性

将方斑东风螺Babylonia areolata(Link)的早期发育从早期面盘幼体到变态后15d的稚贝分成6个阶段,分别测定各阶段蛋白酶(P)、脂肪酶(L)、淀粉酶(A)和纤维素酶(C)活性。研究结果表明,随着幼体发育的推进,蛋白酶比活力逐渐减小,脂肪酶比活力逐渐增加,淀粉酶和纤维素酶比活力则在变态前逐渐增加,而变态后逐渐减小。比较A/P和(A C)/(P L)两种指标指示方斑东风螺幼体发育过程中的食性变化,提出(A C)/(P L)用于指示方斑东风螺食性变化更合理、准确。综合比较幼体发育各阶段4种消化酶活性,发现在面盘幼体向稚贝变态期间,幼体消化能力最弱,原因与幼体的变态过程有关。实验结果也验证了贝类具有分泌纤维素酶的能力,该酶在东风螺对纤维素分解中占主导地位。     

Toxicity of a fuel oil to the eggs of parhyale hawaiensis and amphithoe valida (amphipoda)

Experiments were carried out on the toxicity of a fuel oil to developing embryos and juveniles of two marine amphipods. Parhyale hawaiensis eggs were able to develop from early germinal disc stage (early eggs) to juvenile form in concentrations of 10 (2 ppm) up to 40% (8 ppm) WSF, but deleterious effects were evident in hatching success of eggs and survival of juveniles at concentrations ≥ 10% WSF. The juveniles that hatched from those eggs in higher WSFs had greater mortality than the juveniles that hatched from the eggs exposed to lower WSFs. Survival of juveniles from late eggs (cephalic region differentiated) was also greater than those from early eggs. Amphithoe valida eggs were more sensitive to the WSF of fuel oil than the eggs of P. hawaiensis; < 10% of the test eggs hatched in the 4 ppm WSF compared with 66% of P. hawaiensis eggs at the same concentration. The toxicity of fuel oil to the two amphipod eggs is therefore dependent on the concentration and duration of exposure on the one hand, and on the developmental stages and probably the nature of the egg case on the other.     

The roles of thyroid hormone receptor and T3 in metamorphosis of Haliotis diversicolor

Thyroid hormone is a kind of important hormone which regulates metamorphosis. Its role is well described in amphibian metamorphosis. Thyroid hormones (T3 and T4) have also been demonstrated to play a role in metamorphosis of marine invertebrates. However, the mechanism of thyroid hormone in metamorphosis of marine invertebrates remains unknown. A homolog of vertebrate thyroid hormone receptor (TR) was cloned and identified in abalone Haliotis diversicolor and was named HdTR . The mRNA expressions of HdTR , thyroid peroxidase ( TPO ), thyroid peroxidase 1 ( TPO1 ), idothyronine deiodinase Ⅲ( IDⅢ) and integrin alpha-V ( ITGAV ) had significant diff erence in metamorphosis of H . diversicolor . Metamorphosis rate and mortality rate were significantly diff erent in HdTR RNAi experiment and T3 inducing experiment. In RNAi experiment, ITGAV and CCND1 (cyclin D1) expression of dsRNA HdTR exposing group were significantly lower than those of blank control and negative control. But CTNNB (catenin beta) expression of dsRNA HdTR exposing group was higher than that those of blank control and negative control. ERK (extracellular signal regulated kinases) and PI3K (phosphoinositide-3-kinase) had no significant diff erence in RNAi experiment. Moreover, ITGAV of 1 μmol/L T3 group was significantly lower than that of 0 μmol/L T3 group, PI3K expression of 10 μmol/L T3 group was higher than that of 0 μmol/L T3 group, and the other genes expression had no significant diff erence in T3 inducing experiment. The data of genes expression suggested that CCND1 might be an eff ector gene of TR genomic action, while CTNNB might be regulated by unliganded TR. CCND1 and CTNNB may be involved in cell proliferation of metamorphosis. T3 might regulate the expression level of PI3K via nongenomic way. These results shed light on the mechanism of thyroid hormone in abalone metamorphosis.     

Lipid biomarkers and bacterial lipase activities as indicators of organic matter and bacterial dynamics in contrasted regimes at the DYFAMED site, NW Mediterranean

This study investigated the relationships between dissolved organic matter (DOM) composition and bacterial dynamics on short time scale during spring mesotrophic (March 2003) and summer oligotrophic (June 2003) regimes, in a 0–500 m depth water column with almost no advection, at the DYFAMED site, NW Mediterranean. DOM was characterized by analyzing dissolved organic carbon (DOC), colored dissolved organic matter (CDOM) and lipid class biotracers. Bacterial dynamic was assessed through the measurement of in situ bacterial lipase activity, abundance, production and bacterial community structure. We made the assumption that by coupling the ambient concentration of hydrolysable acyl-lipids with the measurement of their in situ bacterial hydrolysis rates (i.e. the free fatty acids release rate) would provide new insights about bacterial response to change in environmental conditions. The seasonal transition from spring to summer was accompanied by a significant accumulation of excess DOC (+5 μM) (ANOVA, p<0.05, n=8) in the upper layer (0–50 m). In this layer, the free fatty acids release rate to the bacterial carbon demand (BCD) ratio increased from 0.6±0.3 in March to 1.3±1.0 in June (ANOVA, p<0.05, n=8) showing that more uncoupling between the hydrolysis of the acyl-lipids and the BCD occurred during the evolution of the season, and that free fatty acids contributed to the excess DOC. The increase of lipolysis index and CDOM absorbance (from 0.24±0.17 to 0.39±0.13 and from 0.076±0.039 to 0.144±0.068; ANOVA, p<0.05, n=8, respectively), and the higher contribution of triglycerides, wax esters and phospholipids (from <5% to 12–31%) to the lipid pool reflected the change in the DOM quality. In addition to a strong increase of bacterial lipase activity per cell (51.4±29.4–418.3±290.6 Ag C cell⁻¹ h⁻¹), a significant percentage of ribotypes (39%) was different between spring and summer in the deep chlorophyll maximum (DCM) layer in particular, suggesting a shift in the bacterial community structure due to the different trophic conditions. At both seasons, in the chlorophyll layers, diel variations of DOM and bacterial parameters reflected a better bioavailability and/or DOM utilization by bacteria at night (the ratio of free fatty acids release rate to bacterial carbon demand decreased), most likely related to the zooplankton trophic behaviour. In mesotrophic conditions, such day/night pattern was driving changes in the bacterial community structure. In more oligotrophic period, diel variations in bacterial community structure were depth dependent in relation to the strong summer stratification.     

Further investigations on why POC concentrations differ in samples collected by Niskin bottle and in situ pump

Particulate organic carbon (POC) concentrations measured in bottles are often higher than those measured by in situ pumps when samples are taken concurrently. In previous work, we suggested that differential collection of zooplankton might explain this systematic discrepancy in POC between these small volume (bottle) and large volume (in situ pump) techniques. We have now further quantified the carbon contributed by zooplankton collected in the >70-μm particulate fractions from both bottles and pumps at sites in the Mediterranean Sea and Long Island Sound. Our results show that zooplankton abundance and lipid concentrations from zooplankton are one order of magnitude higher in the bottles than in the pumps, supporting the idea that part of the pump–bottle difference is due to collection of more zooplankton by the bottles. Particle washout off the 70-μm mesh used in the in situ pump may cause loss of some particles as well. However, zooplankton in the >70-μm fraction from the bottles contributed only about 1–2 μM POC, which cannot explain the up to 20 μM POC differences observed in this study. Thus, the mechanisms leading to such a large POC difference are still unclear and need to be further investigated. POC concentrations measured using microquartz filters were similar to those using glass fiber filters, suggesting that filter types cannot explain the higher POC observed in bottles, where glass fiber filters are normally used. Furthermore, we investigated several different pump inlet designs to determine how these might affect the ability of pumps to collect and retain large (>70 μm) particles, including zooplankton. The comparison among different pump inlets suggests that inlet design affects the efficiency and retention of large particles and that a sealed filter holder with a narrow right-angle tubular opening is the most efficient at catching/retaining zooplankton.     

Resistance to UV and persulphate oxidation of dissolved organic carbon produced by selected marine phytoplankton

We describe an investigation into the reactivity of dissolved organic carbon (DOC), produced from marine algae, to conventional persulphate and ultraviolet (UV) oxidation methods. Marine algae were grown in batch culture in the presence of ¹⁴C bicarbonate and filtered samples of the phytoplankton dissolved organic carbon (PDOC) were oxidized with persulphate and UV techniques. The amount of fixed label found in solution after the oxidation procedures was compared with the initial amount of labelled DOC. Marine algae examined in this way included: the diatoms, Chaetoceros gracilis, Skeletonema costatum, Phaeodactylum tricornutum; the flagellate, Isochrysis galbana; and the cyanobacterium, Synechococcus strain DC2. It was found that 5–18% of the DOC produced by these phytoplankton resisted persulphate oxidation. Samples were also measured for their resistance to UV oxidation by an autoanalyzer method. It was found that 15–27% of these samples resisted UV oxidation. However, 95% of PDOC was oxidized after exposure for 6 h to high intensity UV irradiation using a variable exposure time system. P. tricornutum and Synechococcus PDOC samples were ultrafiltered into low molecular weight (< 10 000 Da) and colloidal (> 10 000 Da) size fractions. Both species produced mainly (> 68%) low molecular weight material. Slightly greater resistance to persulphate oxidation was generally found for the colloidal Synechococcus PDOC (15–22%) than for the low molecular weight material (14–17%). However, the opposite was found for the P. tricornutum PDOC which generally showed less resistance for the colloidal fraction (5–12%) than for the low molecular weight fraction (10–15%).Experiments were also conducted to determine the effects of short-term (days) and long-term (months) ageing of PDOC solutions in the presence of microbial populations from coastal seawater. Long-term ageing decreased the amount of PDOC that resisted oxidation in all cases. However, the fraction of PDOC resisting persulphate oxidation increased by small amounts over a short-term experiment. Increased resistance was attributed to preferential degradation of biologically and chemically labile components of PDOC by bacteria.The percentages of phytoplankton-produced (and microbially aged) DOC found in this study to resist UV or persulphate oxidation were low (5–27%), compared with those values (50–65%) reported for DOC in surface seawater on the basis of recent high temperature catalytic oxidation analyses.     

Chemical induction of settlement and metamorphosis in two Babylonia (Gastropoda) larvae

INTRODUCTIONStudiesonchemicalinductionofsettlementandmetamorphosisinmarineGastropodalarvaehaveadvancedconsiderablyoverdecades,manyofwhichfocusonHalionsrufescensandPhestillasibogae.AccordingtotheavailabledataItheeffectivefactorswhichcaninduceGastropodalarvaetosettleandmetamorphose,includenatureinducers,neurotransmittersandtheirstructuralanalogs,inorganicion(K )andorganicsolvents.Sincethemechanismofmetamorphosisingastropodlarvaediffersmarkedlyamongvariousspecies,itisnecessarytoexamineandc…     

Pure compound and sediment elutriate embryo larval assay of winter flounder from the Hudson/Raritan Estuary and Long Island Sound

Winter flounder (Pseudopleuronectes americanus), a demersal egg layer, is at risk from dioxins and metals in the sediments of the Hudson/Raritan Estuary (the Estuary). The effects of tetrachlorodibenzo-p-dioxin (TCDD), mercuric chloride and sediment elutriates on the embryos of winter flounder from four sites in the Estuary, and a reference site: Long Island Sound (LI) were examined. Embryos were exposed from 2 days after fertilization until 2 days post hatch. TCDD absorption by embryos/larvae ranged from 13 to 44%, with an average 23.4%. One experiment, with the LI population showed a LD₅₀ of 1.8 pg/larva (2.6 ng/g) and LD₁₀ of 0.9 pg/larva (1.4 ng/g). Major observed lesions included pericardial edema (PE), and spinal lesions (SL). Background lesion occurrence in controls was 0.31 lesions/individual, and 0.27 major lesions/individual. NOELs were observed for PE 0.3–0.5 pg/larva, and SL 1.4–9.0 pg/larva. PE and SL are good indicators of low and high dose effects of TCDD in winter flounder, respectively. With mercuric chloride all populations showed 50% mortality at 100 μg/l, and 50% lesions at 25–50 μg/l. Three of seven sediment elutriates collected from the Estuary were acutely toxic. Site-specific variability in sediment toxicity and breeding stock sensitivity to known contaminants was observed. Future studies should incorporate metamorphosis from larva to juvenile. (USEPA:CR825437, NIEHS:ES07148).     

Changes in serum thyroid hormone levels and thyroid gland activity of artificially maturing female Japanese eel(Anguilla japonica)

In order to evaluate the involvement of the thyroid gland in reproduction, thyroid functioning was described in female cultivated Japanese eel (Anguilla japonica) in progressive stages of sexual maturation induced by chum salmon pituitary homogenate (SPH) treatment. Serum thyroid hormones, thyroxine (T4) and triiodothyronine (T3) were also measured in each stage. Thyroid gland activity (epithelial cell height) was high before SPH injection (previtellogenic stage), further increasing at the early vitellogenic stage, thereafter decreasing to late vitellogenic and migratory nucleus stages. The profiles of both T3 and T4 changed during vitellogenesis, being high during previtellogenic and early vitellogenic stages, and subsequently declining, thus mimicking thyroid gland activity. These results suggest that the thyroid has relation with eel ovarian development during artificial maturation.     

Dynamics of carbohydrates in the Norwegian Sea inferred from monthly profiles collected during 3 years at 66°N, 2°E

Water samples were collected monthly for 3 years at 66°N, 2°E in the Norwegian Sea, 250 nautical miles off the Norwegian coast. Concentrations of mono- and polysaccharides were measured with the 2,4,6-tripyridyl-s-triazine (TPTZ) spectroscopic method. Total dissolved carbohydrates varied from 3.4 to 28.2 μM C of all samples and the ratio of carbohydrate to dissolved organic C (DOC) varied from an average of 14% at 0–25 m depth to 11% at 800–2000 m depth. This indicates that dissolved carbohydrates were a significant constituent of DOC in the Norwegian Sea. Polysaccharides varied from 0.4 to 21.5 μM C and monosaccharides from 0.7 to 11.7 μM C at all depths. The level of monosaccharides was relatively constant at 2.8–3.2 μM C below the euphotic zone, whereas polysaccharides showed more varying concentrations. Dissolved carbohydrates accumulated during the productive season, reaching maximum concentrations during summer although interannual differences were observed. A significant positive correlation between Chl a and soluble carbohydrate was found in one growing season with nutrient analyses. Average values for total carbohydrates were highest in the surface – 0 to 25 m – with 13.3 μM C and decreased to 8.4 μM C at 800–2000 m depth. The ratio of monosaccharides to polysaccharides exhibited a marked seasonal variation, increased from January to a maximum in June of 1.1, and declined to 0.5 in July.     

Effects of β-naphthoflavone on hepatic biotransformation and glutathione biosynthesis in largemouth bass (Micropterus salmoides)

We are investigating the effects of in vivo exposure of prototypical enzyme inducing agents on hepatic biotransformation enzyme expression in largemouth bass (Micropterus salmoides), a predatory game fish found throughout the United States and Canada. The current study targeted those genes involved in biotransformation and oxidative stress that may be regulated by Ah-receptor-dependent pathways. Exposure of bass to β-naphthoflavone (β-NF, 66 mg/kg, i.p.) elicited a 7–9-fold increase in hepatic microsomal cytochrome P4501A-dependent ethoxyresorufin O-deethylase (EROD) activities, but did not affect cytosolic GST catalytic activities toward 1-chloro-2,4-dinitrobenzene (CDNB) or 5-androstene-3,17-dione (ADI). Glutathione S-transferase A (GST-A) mRNA expression exhibited a transient, but non-significant increase following exposure to β-NF, and generally tracked the minimal changes observed in GST–CDNB activities. Expression of the mRNA encoding glutamate-cysteine ligase catalytic subunit (GCLC), the rate-limiting enzyme in glutathione (GSH) biosynthesis, was increased 1.7-fold by β-NF. Changes in GCLC mRNA expression were paralleled by increases in intracellular GSH. In summary, largemouth bass hepatic CYP1A-dependent and GSH biosynthetic pathways, and to a lesser extent GST, are responsive to exposure to β-NF.     

Seasonal dynamics of inorganic nutrients and phytoplankton biomass in the NW Alboran Sea

The seasonal dynamics of inorganic nutrients and phytoplankton biomass (chlorophyll a), and its relation with hydrological features, was studied in the NW Alboran Sea during four cruises conducted in February, April, July and October 2002. In the upper layers, the seasonal pattern of nutrient concentrations and their molar ratios (N:Si:P) was greatly influenced by hydrological conditions. The higher nutrient concentrations were observed during the spring cruise (2.54 μM NO₃⁻, 0.21 μM PO₄³⁻ and 1.55 μM Si(OH)₄, on average), coinciding with the increase of salinity due to upwelling induced by westerlies. The lowest nutrient concentrations were observed during summer (<0.54 μM NO₃⁻, 0.13 μM PO₄³⁻ and 0.75 μM Si(OH)₄, on average), when the lower salinities were detected. Nutrient molar ratios (N:Si:P) followed the same seasonal pattern as nutrient distribution. During all the cruises, the ratio N:P in the top 20 m was lower than 16:1, indicating a NO₃⁻ deficiency relative to PO₄³⁻. The N:P ratio increased with depth, reaching values higher than 16:1 in the deeper layers (200–300 m). The N:Si ratio in the top 20 m was lower than 1:1, excepting during spring when N:Si ratios higher than 1:1 were observed in some stations due to the upwelling event. The N:Si ratio increased with depth, showing a maximum at 50–100 m (>1.5:1), which indicates a shift towards Si-deficiency in these layers. The Si:P ratio was much lower than 16:1 throughout the water column during the four cruises. In general, the spatial and seasonal variation of phytoplankton biomass showed a strong coupling with hydrological and chemical fields. The higher chlorophyll a concentrations at the depth of the chlorophyll maximum were found in April (2.57 mg m⁻³ on average), while the lowest phytoplankton biomass corresponded to the winter cruise (0.74 mg m⁻³ on average). The low nitrate concentrations together with the low N:P ratios found in the upper layers (top 20 m) during the winter, summer and autumn cruises suggest that N-limitation could occur in these layers during great part of the year. However, N-limitation during the spring cruise was temporally overcome by nutrient enrichment caused by an intense wind-driven upwelling event.     

Distribution and sinking rates of phytoplankton, detritus, and particulate biogenic silica in the Laptev Sea and Lena River (Arctic Siberia)

Concentrations and sinking rates of particulate biogenic silica (BSi), chlorophyll a (chl a) and phaeopigments (phae) (< 3 μm, 3–10 μm, > 10 μm and total), as well as the abundances of the major phytoplankton species, were studied during September 1991 in the Eastern Laptev Sea and the lower Lena River (Siberian Arctic). The highest chl a concentrations were found in two major “new” production regimes of the study area: (1) a deep chl a maximum (5.8 mg chl a m⁻³) (formed by the diatom Chaetoceros socialis) at 30 m depth on the outer shelf of the northern Laptev Sea, and (2) in the Lena River, where the phytoplankton community was dominated by fresh water diatoms (1.5 to 4.5 mg chl a m⁻³). Elevated chl a concentrations were also found in the river plume phytoplankton community (dominated by brackish water diatoms), NE of the Lena delta. In the Laptev Sea, the low chl a (0.1 to 3 mg chl a m⁻³) and high phae concentrations (0.5 to 14 mg phae m⁻³) indicated that the phytoplankton community (dominated by picoplanktic algae and nanoflagellates) was already senescent and affected by grazing losses. Biogenic silica values were highest in the Lena River (4 to 17 μM) as compared to the low values found in the Laptev Sea (0.3 to 4 μM). The large chl a size fraction, phae and BSi in the Lena River samples revealed the highest measured sinking rates (1.4, 2.3, and 1.5 m d⁻¹, respectively). The formation of a strong halocline, decreasing turbulence, and possible nutrient deficiency resulted in death, disintegration and rapid sedimentation of fresh water diatoms. This was accompanied by a decrease in the BSi concentration and growth of the picoplanktic size fraction (< 3 μm) in the estuarine mixing zone (Gulf of Buorkhaya). Only a minor part of BSi was bound to intact diatom cells (< 3%) in the surface layer, most of which being apparently associated with detrital particles. In the Lena River, approximately 12% of the total silica was bound to BSi fraction, yet elsewhere in the Laptev Sea and in the estuarine mixing zone the BSi:total silica ratio was ≤ 5%. Thus, the results reflected the successional stage of a late summer phytoplankton community, characterized by dominance of small autotrophs and patchy distribution of senescent diatoms no longer able to affect the relative high levels of dissolved silica supplied by the Lena River.     

Antagonistic effects of xenobiotics on steroid-induced final maturation of Atlantic croaker oocytes in vitro

The antagonistic effects of Kepone (chlordecone) and o,p'-DDD on maturation-inducing steroid (17α,20β,21-trihydroxy-4-pregnen-3-one, 20β-S) induced final maturation of Atlantic croaker (Micropogonias undulatus). Oocytes were investigated in an in vitro germinal vesicle breakdown (GVBD) bioassay. Exposure to Kepone or o,p'-DDD (100 nM-100 μM) prevented the majority of the oocytes from completing GVBD in response to 20β-S; many of them were arrested at the lipid coalescence or germinal vesicle migration stages when exposed to high xenobiotic concentrations (100 μM). In addition, clearing of the ooplasm, oil droplet formation and hydration were incomplete in xenobiotic-exposed oocytes that underwent GVBD. The xenobiotics inhibited GVBD in a concentration-dependent manner. One minute exposure to either of the xenobiotics was sufficient to block GVBD. Washing the follicle-enclosed oocytes after xenobiotic exposure restored their ability to undergo GVBD in response to 20β-S. These results suggest that o,p'-DDD and Kepone exert antagonistic effects on 20β-S action, possibly by competing for the ovarian 20β-S membrane receptor.     

海湾扇贝幼虫变态过程中体内神经递质含量的变化

采用生化方法测定了不同发育阶段及人工诱导后海湾扇贝 (Argopectenirradians)幼虫体内去甲肾上腺素 (NE)、多巴胺 (DA)和 5 羟色胺 ( 5 HT)含量的变化。结果表明 ,海湾扇贝幼虫体内去甲肾上腺素含量在变态前后没有明显变化 ,变态前为 2 35 2pg/mg湿重 ,变态后为2 770pg/mg湿重。多巴胺和 5 羟色胺含量在变态前 (第 1 3天 )急剧增加 ,比第 1 2天的幼虫分别增加了 2 .8倍和 4.7倍 ,变态后急剧下降 ,变态后幼苗比第 1 3天的幼虫分别降低了 2 5 .1倍和 1 6.4倍。海湾扇贝幼虫体内DA∶NE比值和 5 HT∶NE比值在变态前后变化剧烈。DA∶NE比值和 5 HT∶NE比值在变态前 (第 1 3天 )急剧增加 ,比第 1 2天的幼虫增加了 3.0倍 (DA∶NE)和 5 .0倍 ( 5 HT∶NE) ;变态后急剧降低 ,变态后幼苗比第 1 3天的幼虫降低了 2 9.8倍 (DA∶NE)和 1 9.5倍 ( 5 HT∶NE)。海湾扇贝幼虫经氯化钾和氯化钙诱导 2 4h后 ,体内去甲肾上腺素、多巴胺和 5 羟色胺以及DA∶NE比值和 5 HT∶NE比值均有所降低。本实验的结果表明 ,多巴胺和 5 羟色胺可能启动了海湾扇贝幼虫的变态过程。     

Effects of water-borne copper on metallothionein and lipid peroxidation in the marine amphipod gammarus locusta

The aim of the current work was to determine over 10 days the effects of water-borne exposure of the marine amphipod Gammarus locusta to 4 μgCu l⁻¹ on the metallothionein (MT; measured by differential pulse polarography) protection system and lipid peroxidation (LP; thiobarbituric acid-reactive malondialdehyde equivalents) as a measure of oxidative damage. MT levels in exposed animals increased significantly at day 2 (36% > control; P<0.001) and remained high at days 6 and 10 (55 and 38%, respectively, P<0.001). The maximum level of MT at day 6 coincided with the highest Cu body-burden. LP increased within 1 day of exposure, indicative of Cu as an oxidative stressor. However, in contrast to MT, the highest LP level was seen at day 4 (68% > control, P<0.001) before returning to control values by day 6, indicating a protective role of MT against the pro-oxidant effects of Cu.     

Significance of biofilm proteins in modulating cyprid metamorphosis of Balanus amphitrite (Cirripedia: Thoracica)

The role of biofilm proteins in modulating cyprid metamorphosis is not well understood. In the present investigation this possibility was explored by disrupting biofilm proteins using commercially available protease. The influence of natural biofilm and artificial biofilms of Aeromonas salmonicida salmonicida and Bacillus brevis and their culture supernatants and exopolysaccharides obtained under different nutritional conditions was evaluated. Natural biofilm facilitated cyprid metamorphosis in Balanus amphitrite, whereas monospecies biofilms did not. The treatment of biofilms with protease resulted in a decline in metamorphosis, except in B. brevis, where although metamorphosis was negated initially, it increased with the protease concentration and observation days. As the response of cyprids to exopolysaccharides produced by A. salmonicida salmonicida was dependent on protein concentration, protease treatment resulted in a decrease in metamorphosis, whereas in case of B. brevis biofilms, protease hydrolysis products could be important in providing settlement cues. Thus, biofilm bacterial components play an important role in the cyprid metamorphosis of Balanus amphitrite.     

Species and concentrations of selenium and nutrients in Tanabe Bay during red tide due toGymnodinium nagasakiense

A red tide due toGymnodinium nagasakiense was observed in August 1988 in Tanabe Bay, Wakayama Prefecture, Japan. The maximum cell concentration ofG. nagasakiense reached 1×10⁵ cells ml^–1 at the surface water. From May to September 1988, the following were monitored: water temperature, salinity, chlorophylla, D.O., dissolved nutrients (NO₂–N, NO₃–N, NH₄–N, PO₄–P DON, DOP), particulate nutrients (PON, POP) and three dissolved selenium species [Se(IV), Se(VI), Organic Se]. Dissolved inorganic nitrogen (NO₃–N, NH₄–N) decreased but PON, POP, DON, DOP and inorganic phosphate increased at the peak of the bloom. The concentration of organic selenium increased up to the bloom initiation period which started on 5 July, and then the concentration of Se(IV) increased as the concentration of organic selenium decreased at the peak of the bloom (3 August). The strong relationship was found between the concentration of Se(IV) and the cell concentration ofG. nagasakiense (r ²=0.98). The Se(IV) requirement ofG. nagasakiense was 2.89×10^–17 moles cell^–1, which was agreed well with 4.4×10^–17 moles cell^–1 found in a laboratory experiment onG. nagasakiense using selenium spiked artificial sea water medium. The average ratio of Se(IV) to dissolved inorganic nitrogen (DIN) during the red tide bloom was 11441, the ratio of Se(IV) to DIN at the surface with the maximum cell concentration ofG. nagasakiense of 1×10⁵ cells ml^–1 was 1137. These results suggested that selenium may play an important role in red tide outbreak ofG. nagasakiense.     

Determination of adsorbed and absorbed polychlorinated biphenyls (PCBs) in seawater microorganisms

To distinguish between adsorbed and absorbed PCBs in seawater microorganisms, 2,2′,4,4′,5,5′-hexachlorobiphenyl (IUPAC #153; HCB) was added to a pure culture of Chrysochromulina apheles Moestrup et H.A. Thomsen. After the addition of the HCB, the cells were immediately harvested onto 2 μm polycarbonate filters and rinsed with a gradient of ethanol concentrations. Rinsing with 40% ethanol (v/v) was found to remove 80% of the HCB, which was loosely adsorbed to the cell surfaces, but did not extract the interior of the cells, as tested by chlorophyll a analysis. This method was used in a time course experiment which estimated PCBs adsorption and absorption to different groups of plankton organisms. Three different ¹⁴C-PCBs, 4-chlorobiphenyl (IUPAC #3; MCB), 2,2′,5,5′-tetrachlorobiphenyl (IUPAC #52; TCB), and 2,2′,4,4′,5,5′-hexachlorobiphenyl (IUPAC #153, HCB), were incubated in seawater from the northern Baltic Sea during a spring bloom. Samples were taken every third day and separated by filtration into three fractions; 0.2–2 μm (bacteria), 2–10 μm (flagellates), and >10 μm (microplankton; phytoplankton and protozoa). Two subsamples were retained from each size fraction. One of the subsamples was left untreated, to obtain adsorbed plus absorbed PCB, while the other subsample was rinsed with 40% of ethanol, to obtain the absorbed PCB. The sorption was found to vary depending on the hydrophobicity of the compounds, the structure of the cell membranes, and the lipid content and composition of the cells. The absorption increased for the TCB and the HCB in the largest size fraction over time, which coincided with an increase of the neutral and non-polar lipids.