Molecular cloning of superoxide dismutase (Cu/Zn-SOD) from aquatic molluscs

The potential of the first line of the active oxygen-scavenging system, partial cDNA encoding Cu/Zn superoxide dismutase (SOD) was isolated in three aquatic mollusc species: Ruditapes decussatus (marine clam), Dreissena polymorpha (continental water mussel) and Bathymodiolus azoricus (hydrothermal vent mussel). These SOD cDNA fragments were amplified by PCR with degenerate oligonucleotide primers derived from the amino acid sequence conserved in the Cu/Zn-SOD from several other organisms. A partial cDNA of CuZn-SOD was obtained for R. decussates (510 bp), D. polymorpha (510 bp) and B. azoricus (195 bp). The deduced amino acid sequence showed high similarity among the three mollusc species (57-63%) and among other species (50-65%). The residues involved in coordinating copper (His-47, 49, 64, 121) and zinc (His-64, 72, 81 and Asp-84) were well conserved among the three Cu/Zn-SOD sequences.     

Tissue Cu, Fe and Mn concentrations in different-aged and different functional thallus regions of three brown algae from western Ireland

Copper and iron concentrations in three brown algae, Ascophyllum nodosum, Fucus vesiculosus and Laminaria digitata (and additionally Mn in L. digitata) from the Irish west coast were determined using flame atomic absorption spectrophotometry. Metal concentrations in the three species were indicative of prevailing bioavailable metal concentrations in situ but varied greatly between functional tissue parts, between sites and over time. Cu concentrations in actively growing tips of A. nodosum decreased over a 4-month period during autumn/winter, while Fe concentrations increased. Both Fe and Cu concentrations in different thallus sections of A. nodosum and F. vesiculosus increased with increasing age of thallus part in a clean site, but there was no consistent trend for F. vesiculosus from an industrialized site. Within sites, concentrations of all Cu and Fe were similar in both fucoids, but concentrations at the industrialized site were about twice as high as at the pristine site. In L. digitata, all three metals were highest in holdfasts, but had distinctly different distribution patterns in stipes and blade sections, which were most likely related to growth pattern and tissue function. Fe was lowest in meristematic and young blade regions, suggesting small-scale Fe limitation in actively growing tissue. Mn concentrations were higher in distal blade sections than in stipes, and Cu concentrations were highest in meristematic and young thallus parts.     

The impact of musk ketone on reproduction in zebrafish (Danio rerio)

Nitro musks are a group of nitrated benzenes mainly used by the fragrance industry as a substitute for natural musk. Two of the most common nitro musks, musk xylene and musk ketone, have been detected in water, fish, human adipose tissue, as well as in human breast milk. In this study, female zebrafish were dietary-exposed to musk ketone, prior to spawning with unexposed males. The fish were exposed for 8 weeks at two different dose levels. Accumulation of the compound in the fish as well as reproductive success were studied. Exposed females had reduced body weight and length, as well as reduced liver- and gonad somatic index. The results from the reproduction study showed a dose-dependent reduction in fecundity. Early life-stage mortality was increased and the median survival time was reduced. In addition, an embryo/larvae toxicity test was performed, using newly fertilized zebrafish eggs from unexposed parental fish. Eggs were exposed to a series of different concentrations of musk ketone via the surrounding water. A NOEC value of 10 μg/l and a LOEC value of 33 μg/l was determined. Conclusively, the present study clearly shows that musk ketone negatively affects reproduction and early life-stage survival in zebrafish.     

养殖水体中氨氮对大菱鲆的急性毒性效应研究

为了评估大菱鲆(Scophthalmus maximus)耐氨氮性状的遗传参数和为循环水养殖大菱鲆提供科学指导,作者研究了养殖海水中氨氮对4月龄和8月龄大菱鲆的急性毒性效应,同时对比了氨氮对白化和正常个体、雌性和雄性个体的急性毒性效应差别。结果表明:非离子氨对4月龄大菱鲆的24、48、72、96 h LC50分别为2.19、1.94、1.80、1.72 mg/L,对8月龄大菱鲆的24、48、72、96 h LC50分别为3.64、3.02、2.93、2.86 mg/L,非离子氨对4月龄和8月龄大菱鲆的安全质量浓度分别为0.17和0.29 mg/L;氨氮对白化和正常大菱鲆以及雌性和雄性大菱鲆的急性毒性效应均没有显著性差异,研究结果为进一步选育大菱鲆的耐氨氮品系提供科学依据。     

Influence of combined changes in salinity and copper modulation on growth and copper uptake in the tropical green macroalga Ulva reticulata

The influence of salinity on growth and Cu uptake in the green macroalga Ulva reticulata collected from the intertidal area in the Western Indian Ocean was studied under controlled laboratory conditions. Exposure concentrations ranged from 5 to 500 μg Cu l⁻¹ at five salinities (ranging 20–40). The accumulation of Cu increased with decreasing salinity, so that the uptake at 500 μg Cu l⁻¹ was approximately 2.7, 2.4 and 2.0 times higher at salinities of 20, 25, and 30 respectively, than uptake at salinity of 35, and with uptake being lowest at salinity of 40. Ulva maintained a positive growth rate over the whole salinity range (20–40), with highest rates at salinity of 35. When exposing to Cu at low salinities (20 and 25), the growth rate of Ulva was strongly inhibited suggesting an increase in toxicity of Cu with decreasing salinity. EC50 and NOEC increased with increase in salinity, implying a reduced Cu toxicity at high salinities. It was concluded that salinity needs to be considered when using macroalgae, such as U. reticulata, as a bioindicator of heavy metals in areas with heavy rainfall, underground fresh water intrusion or in estuaries, as they might accumulate more metals and be more negatively affected.     

Histological and morphological alterations induced by copper exposure in Laeonereis acuta (Polychaeta, Nereididae)

Laeonereis acuta (Polychaeta, Nereididae) was collected in an unpolluted (UP) and an polluted (P) site at the Patos Lagoon estuary (Southern Brazil) and maintained under control conditions (UPC and PC, respectively) or exposed to waterborne copper (UPCu and PCu; 500 μg Cu/l), for 48 h. Four groups (aaUPC, aaPC, aaUPCu, and aaPCu) were also pre-exposed for 48 h to ascorbic acid (aa; 0.1 mM) before copper exposure. Histological and morphological alterations, as well as oxygen consumption changes were evaluated. Independently of the sampling site and the pre-exposure to the ascorbic acid, morphological abnormalities were evident in more than 80% of worms exposed to copper. Conspicuous histological changes (coeloma obliteration, cuticle separation from the epidermis, and absence of dorsal vessel) were also observed. In addition, PCu worms showed loss of the digestive epithelium and coiling behavior. Similar oxygen consumption values were observed in control and copper exposed worms.     

Arm regeneration frequency in eight species of ophiuroidea (Echinodermata) from European sea areas

Eight ophiuroid species, six from the northern Kattegat-eastern Skagerrak and three from the northern Adriatic Sea, were examined for regeneration of arms. The species were separated into groups based upon mode of feeding and habitat. Comparison between groups collected in the northern Kattegat-eastern Skagerrak showed that infaunal suspension- and deposit-feeding species (Amphiura filiformis and A. chiajei) had significantly more scars per arm (mean number 0.78) than epibenthic suspension feeders (Ophiothrix fragilis and Ophiocomina nigra, 0.29) or epibenthic carnivores and deposit feeders (Ophiura ophiura and O. albida, 0.13). Spatial variation in arm regeneration incidence was found between sampling sites in the northern Kattegat-eastern Skagerrak for Amphiura filiformis and in the northern Adriatic Sea for Ophiothrix quinquemaculata. The ash-free dry weight (AFDW) and nitrogen (N) contents were measured in arms of six species of brittle-stars from the northern Kattegat-eastern Skagerrak. Differences between species were found, with highest concentrations of AFDW and N in Amphiura filiformis, intermediate in A. chiajei, Ophiocomina nigra and Ophiothrix fragilis, and lowest in Ophiura ophiura and O. albida. As the infaunal suspension- and deposit-feeding brittle-stars (Amphiura spp.) had the highest proportions of damaged arms and highest AFDW and N contents in their arms in this comparison, it is suggested that selective cropping of arms by demersal fish is the main cause of arm damage on Amphiura spp. in this area.     

Feeding ecology of five commercial shark species of the Celtic Sea through stable isotope and trace metal analysis

In order to trace their feeding habits, stable carbon and nitrogen isotope ratios (delta15N and delta13C), as well as trace metal concentrations (Zn, Cd, Fe, Cu, Se and Hg) were analysed in the tissues of five commercial shark species from the Celtic Sea: the tope shark Galeorhinus galeus, the black-mouthed catshark Galeus melastomus, the starry smooth hound Mustelus asterias, the spiny dogfish Squalus acanthias and the lesser-spotted dogfish Scyliorhinus canicula. Our results were compared to previously described stomach contents and isotopic composition of potential preys. Isotopic ratio delta15N suggested that tope sharks fed at a higher trophic level (16.7 per thousand in the muscle) than the other species, reflecting its piscivorous diet. The lower values of spiny dogfish (11.6 per thousand in the muscle) might be explained, amongst other things, by either its migratory behaviour or its preference for preys from lower trophic levels. Cd and Hg were correlated with isotopic ratios delta13C and delta15N, and were shown to be diet-related whereas Zn, Fe and Cu seemed much more linked to species-specific metabolism. Although this multidisciplinary approach is revealed as a useful tool for the study of shark ecology, the lack of known trophic fractionation suggests that isotopic data be compared to traditional diet analyses.     

改性粘土对虾夷扇贝(Mizuhopecten yessoensis)体内麻痹性贝毒的影响

目前,改性粘土技术已成为国内外普遍认可的有害赤潮治理方法,但在利用该技术治理产毒藻赤潮过程中,对底栖滤食性贝类体内毒素累积和排出的影响尚不清楚。通过室内实验,考察了聚合氯化铝改性粘土(MC I)絮凝典型产毒藻——太平洋亚历山大藻(Alexandrium pacificum)后,虾夷扇贝食用组织和消化腺组织中麻痹性贝毒(paralytic shellfish poisoning,PSP)含量、组分的变化情况。研究结果表明,在前期毒素累积阶段,虾夷扇贝暴露于A. pacificum 3 h后,未添加改性粘土的对照组中虾夷扇贝食用组织毒素水平由空白组的未检出迅速升高至13.24µg STXeq/kg,消化腺组织毒素水平由0.68µg STXeq/kg升高至42.97µg STXeq/kg,第4 d时对照组食用组织、消化腺组织毒素水平达到最高,分别为258.67和3 208.40µg STXeq/kg。添加0.2 g/L MC I的实验组中,3 h后水体中97%的A.pacificum藻细胞被絮凝沉降,此时虾夷扇贝食用组织和消化腺组织毒素水平远低于对照组,分别为6.33和18.39µg STXeq/kg,第2 d时达到98.92和574.54µg STXeq/kg,均显著低于对照组(P<0.05),随后呈下降趋势。在随后的毒素排出阶段,实验组虾夷扇贝食用组织和消化腺组织毒素水平显著低于对照组(P<0.05)。另外,在累积阶段,对照组和实验组虾夷扇贝食用组织和消化腺组织中PSP各组分相对含量差异不大,说明改性粘土未对虾夷扇贝体内的PSP转化造成明显影响。由实验结果可见,改性粘土可有效絮凝去除水体中的A.pacificum藻细胞,减少虾夷扇贝对其的滤食,从而降低了PSP在贝类体内的积累,该研究结果将为改性粘土治理底栖贝类养殖水体有毒赤潮提供科学依据。     

Acute toxicity of heavy metals, tributyltin, ammonia and polycyclic aromatic hydrocarbons to benthic amphipodgrandidierella japonica

Benthic amphipod,Grandidierella japonica widely inhabits the Korean coastal waters and is developed as a standard test species for sediment toxicity tests. We exposedG . japonica to various pollutants including 4 kinds of inorganic metals (Ag, Cd, Cu and Hg), tributyltin [TBT], ammonia and 7 polycyclic aromatic hydrocarbon (PAH) compounds (acenaphthene, chrysene, fluoranthene, fluorene, naphthalene, phenanthrene and pyrene) to estimate the no observed effect concentration (NOEC) and the median lethal concentration (LC50) of each pollutant during the 96-hour acute exposure. Among all tested pollutants, TBT was most toxic toG. japonica, and Hg was most toxic among inorganic metals. The toxicity of pyrene toG. japonica was greatest among PAH compounds, followed by fluoranthene, phenanathrene, acenaphthene, fluorene and naphthalene. The toxicity of PAH compounds was closely related to their physico-chemical characteristics such as K_ow and water solubility.G. japonica responded adequately to pollutant concentrations and exposure durations, and the sensitivity ofG. japonica to various inorganic and organic pollutants was generally comparable to other amphipods used as standard test species in ecotoxicological studies, indicating this species can be applied in the assessment of environments polluted by various harmful substances.     

Recent trends in organochlorine residues in mussels (Mytilus edulis) from the Mersey Estuary.

Blue mussels (Mytilus edulis) taken from several sites in the Mersey Estuary, an urban-industrial water body in NW England, have been analyzed for residues of the persistent organochlorines, DDT, PCB congeners nos. 28, 52, 101, 118, 138, 153 and 180 (ICES7), and alpha-MHCH (alpha-methyl hexachlorocyclohexane). The concentration range for sigmaPCB (ICES7) was 13.9-34.9 microg kg(-1) in 1994 compared with 9.6-31.9 microg kg(-1) in 1998, whilst the equivalent concentrations of sigmaDDT were 8.9-32.4 microg kg(-1) and 4.5-16.8 microg kg(-1), respectively. The concentration of alpha-MHCH, an organochlorine largely restricted to the Mersey Estuary, was 1.2-11 microg kg(-1) in 1994 and 0.3-1.3 microg kg(-1) in 1998. At both sampling dates the inner estuary sites of Rock Ferry and Egremont were more contaminated than the outer estuary sites of New Brighton. Dove Point and Caldy Blacks. The lower concentrations of organochlorines in mussels in 1998 compared with 1994 are thought to be early evidence of significant improvements in water quality derived from major capital investment in the treatment of industrial effluents and sewage wastewater, linked to a combination of stricter legislative controls and the environmental benefits from the adoption of new, clean technologies in manufacturing.     

Cd2+和Cu2+对泥蚶的急性毒性和联合毒性试验

为了解重金属Cd2+和Cu2+对泥蚶的毒害程度和泥蚶对重金属Cd2+和Cu2+的解毒能力,用毒理学实验方法研究了Cd2+和Cu2+对泥蚶(Tegillarca granosa)的急性毒性和联合毒性效应。结果表明:Cd2+、Cu2+对泥蚶的96 h半致死质量浓度分别为6.189、0.460 mg/L;安全质量浓度分别为0.062、0.005 mg/L;相对毒性Cu2+Cd2+。在1︰1毒性单位的联合作用下,Cd2+和Cu2+对泥蚶96 h的半致死质量浓度分别为1.984、0.147 mg/L;安全质量浓度分别为0.020、0.001 mg/L。2种重金属离子的联合毒性大于任一重金属离子的毒性,联合毒性表现为协同作用。     

Nursery areas of red mullet (Mullus barbatus), hake (Merluccius merluccius) and deep-water rose shrimp (Parapenaeus longirostris) in the Eastern-Central Mediterranean Sea

The spatial pattern of the nursery areas of red mullet (Mullus barbatus), hake (Merluccius merluccius) (Linnaeus, 1758) and deep-water rose shrimp (Parapenaeus longirostris) (Lucas, 1846) was studied in the South Adriatic and North Ionian Seas (Eastern-Central Mediterranean) applying geostatistical techniques and data from time series trawl surveys conducted in the area. The analysed variables were: R (number of recruits/km²) and R/Tot (fraction of recruits on the total sampled population). The structural analysis showed a spatial pattern of both variables characterized by continuity on a small scale. Predictions of nursery area localization with probability of finding recruits at different threshold values were obtained through median indicator kriging. For the red mullet the nurseries were mainly identified in the South Adriatic Sea off the Gargano peninsula and between Molfetta and Monopoli within 50 m in depth. The main concentration of hake juveniles was found to be between 100 and 200 m in depth along the Gargano peninsula and between Otranto and Santa Maria di Leuca, where a nursery of deep-water rose shrimp was also detected. An overlapping depth, between 100 and 200 m, was identified for hake and deep-water rose shrimp nurseries. Protection of these areas through limitations of fishing pressure is discussed.     

Metallothioneins and cytosolic metals in Neomysis integer exposed to cadmium at different salinities

In the present study the induction of metallothioneins (MTs) and its relation to cytosolic metal concentrations (Zn, Cu and Cd) in the euryhaline crustacean Neomysis integer exposed to Cd at different salinities was studied. N. integer was exposed to the same free cadmium ion activity of 5.74 x 10(-9) mol l(-1) (i.e. 1/5 of the 96 h LC(50) value expressed as cadmium activity) in hypo-osmotic (5 psu), isosmotic (16 psu) and hyper-osmotic media (25 psu) for 7 days. In this way, the effect of salinity on cadmium speciation was eliminated and therefore the physiological effect of salinity on Cd accumulation and MT induction could be studied. The accumulation of cytosolic Cd in N. integer changed with salinity from 1.11+/-0.05 micromol l(-1) at 5 psu up to 1.43+/-0.17 micromol l(-1) at 25 psu. This could indicate that the physiological response of euryhaline estuarine invertebrates like N. integer to salinity changes can influence the rate of trace metal uptake from solution. While the salinity changes did not cause significant differences in cytosolic Zn concentrations (mean value of all tested salinities: 34.4+/-2.8 micromol l(-1)), an inverse relationship between salinity and cytosolic Cu concentration was observed. The highest concentration of 15.7+/-2.3 micromol Cul(-1) was determined at 5 psu and the lowest 10.9+/-1.4 micromol Cul(-1) at 25 psu. This could point to a possible relationship between the copper concentration and the hemocyanin metabolism in N. integer. This is the first time that differential pulse voltammetry method was applied to MT assays with N. integer. Although the exposure to Cd resulted in a higher Cd cytosolic concentration, no subsequent MT increase was detected. The significant positive correlation between MT levels and cytosolic Cu concentrations (Spearman correlation coefficient r(s)=0.356, p=0.009) implies a strong relationship between MT and Cu in N. integer.     

钦州湾桐花树群落沉积物及植物体中重金属元素累积与分异研究

Mangrove wetlands can reduce heavy metal pollution by trapping heavy metals. In this study, the concentration,transport and bioaccumulation of Cr, Cd, Cu, Zn and Pb in the sediments and different parts of Aegiceras corniculatum at four different sites in the Qinzhou Bay in southwestern China were investigated. The results showed that although the potential ecological risk of all five heavy metals was slight, the concentration of Cr was at a moderate pollution level due to the emissions of industries and aquaculture waste water. Core sediment records indicated that the concentrations of heavy metals at the depth of 0–20 cm were relatively high, showing an increasing trend of heavy metals over the past 20–30 years. Cr, Cu, Pb and Cd accumulated mainly in the roots of A. corniculatum, while Zn accumulated mainly in the stems. Aegiceras corniculatum showed the strongest transport capacity for Zn and Cu and the strongest bioaccumulation ability for Cd. Compared with other mangrove communities, A. corniculatum can be chosen as a restoration species in tropical and subtropical coastal zones polluted by Zn, Cu and Cd.     

Cu2+对泥蚶血红蛋白(Tg-HbⅡ)的过氧化物酶活性与结构的影响

本文利用Cu2+处理泥蚶血红蛋白（Tg-HbⅡ），分别检测Cu2+对Tg-HbⅡ的过氧化物酶活性及其结构的影响，同时检测Pro对Cu2+作用下Tg-HbⅡ的保护作用。结果表明:随着Cu2+浓度的增加，Tg-HbⅡ的过氧化物酶活性逐渐丧失，Cu2+抑制Tg-HbⅡ的过氧化物酶活性类型属于非竞争-竞争混合型抑制，其失活速率具有浓度依赖性，失活过程呈单相变化，表现为一级反应动力学，并且不同浓度Cu2+能影响Tg-HbⅡ的三级结构变化；另外，Pro对Tg-HbⅡ的空间结构及其过氧化物酶活性具有保护作用，能阻止Tg-HbⅡ的聚沉、恢复Tg-HbⅡ的结构与过氧化物酶活性。     

Short-term consequences of a benthic cyanobacterial bloom (Lyngbya majuscula Gomont) for fish and penaeid prawns in Moreton Bay (Queensland, Australia)

This study examined the phenology and ecological consequences of a benthic filamentous cyanobacterial bloom (Lyngbya majuscula) in Deception Bay (Moreton Bay, Queensland, Australia). Bloom initiation occurred in mid December 1999 and expanded to encompass an 8 km² area by April 2000. Small fish and penaeid prawns (<25 cm total length) were quantitatively sampled through periods designated as before, during and after the bloom using a combination of pop-netting within mangroves and beam trawling over adjacent seagrass beds. Data on larger-bodied fish were compiled from daily fishing logs provided by local commercial fishers. Changes in dry mass of bloom material caught in nets and changes in water chemistry were also measured. Mean concentrations of ammonia-N in residual water within mangroves were several orders of magnitude higher in the affected area than in the control and dissolved oxygen was markedly lower in affected areas. Across the study area, mean density, live mass and number of species declined during the bloom, with fish assemblages using mangroves showing greater decline than assemblages using seagrasses. Response at the species level was highly variable; generally, epibenthic species showed a more sustained decline than demersals. Mean monthly fish catch was significantly lower in bloom than non-bloom years. This study has also demonstrated that throughout the bloom, the affected area continued to support a highly diverse and abundant fish and prawn assemblage, and probably maintained its function as an important nursery habitat for many species.     

Influence of sample manipulation on contaminant flux and toxicity at the sediment-water interface

Toxicities of sediments from San Diego and San Francisco Bays were compared in laboratory experiments using sea urchin (Strongylocentrotus purpuratus) embryos exposed to pore water and at the sediment-water interface (SWI). Toxicity was consistently greater to embryos exposed at the SWI to intact (unhomogenized) sediment samples relative to homogenized samples. Measurement of selected trace metals indicated considerably greater fluxes of copper, zinc, and cadmium into overlying waters of intact sediment samples. Inhibition of sea urchin embryo development was generally greater in sediment pore waters relative to SWI exposures. Pore water toxicity may have been due to elevated unionized ammonia concentrations in some samples. The results indicate that invertebrate embryos are amenable to SWI exposures, a more ecologically relevant exposure system, and that sediment homogenization may create artifacts in laboratory toxicity experiments.     

针胞藻海洋卡盾藻(Chattonella marina)和硅藻中肋骨条藻(Skeletonema costatum)藻际细菌的分离、鉴定及影响研究

对实验室培养的两株海洋微藻海洋卡盾藻和中肋骨条藻的可培养细菌进行了分离鉴定，分别在海洋卡盾藻和硅藻中肋骨条藻不同生长时期分离得到48株和34株可培养细菌，去除重复序列后得到12株不同的菌株。这些菌株分属α-变形杆菌纲、γ-变形杆菌纲、拟杆菌和放线菌。α-变形杆菌纲在两种藻的藻际环境中占据优势，其中红杆菌科是最为常见的科，中肋骨条藻的藻际细菌比海洋卡盾藻更为多样化。海洋卡盾藻生长后期细菌密度大幅度上升，可能导致了藻细胞的衰亡。中肋骨条藻相关细菌中有一株对旋链角毛藻的生长具有明显抑制作用，而大部分海洋卡盾藻相关细菌对角毛藻的生长具有抑制作用。本研究结果说明海洋卡盾藻的藻际细菌也许对其与硅藻的种间竞争中具有一定作用。     

Seasonal variation in the concentrations of nutrients in two green macroalgae and nutrient levels in sediments in the Rías Baixas (NW Spain)

Seasonal monitoring of the levels of carbon, nitrogen and phosphorus in two green macroalgae (Ulva and Enteromorpha) was carried out at 22 sampling sites in four embayments (rías) on the northwest coast of Spain. Sediments were also analysed to determine the concentrations of these elements as well as organic matter, organic carbon, iron and texture. In addition, accumulations of Ulva were monitored. Nitrogen levels in algae were similar to those found in comparable studies, whereas carbon concentrations were generally higher and those of phosphorus were lower. As a result the macroalgae were relatively enriched by carbon and nitrogen compared with phosphorus. Seasonal variations in algal tissue nitrogen and phosphorus followed the usual pattern for temperate zones, with minimum levels in summer and maximum levels in winter. Variations in carbon concentrations were much less accentuated. The low levels of P in the algae appear to indicate limitation by this element, however, comparison of the monthly changes in nitrogen and phosphorus in Ulva with the critical concentrations of these elements suggests dual nutrient limitation in this alga in summer. By contrast, the sediments were more enriched by phosphorus; high retention of this element by the sediment may explain the low levels in algae. The highest accumulations of Ulva occurred in spring and summer; hydrodynamic factors appeared to be important in determining the extent of these accumulations.