Carbon dioxide use by chemoautotrophic endosymbionts of hydrothermal vent vestimentiferans: affinities for carbon dioxide, absence of carboxysomes, and δ13C values

The hydrothermal vent vestimentiferans Riftia pachyptila Jones, 1981 and Ridgeia piscesae Jones, 1985 live in habitats with different abundances of external CO₂. R. pachyptila is found in areas with a high input of hydrothermal fluid, and therefore with a high [CO₂]. R. piscesae is found in a range of habitats with low to high levels of hydrothermal fluid input, with a correspondingly broad range of CO₂ concentrations. We examined the strategies for dissolved inorganic carbon (DIC) use by the symbionts from these two species. R. pachyptila were collected from the East Pacific Rise (9°50′N; 104°20′W) in March 1996, and R. piscesae were collected from the Juan de Fuca Ridge (47°57′N; 129°07′W) during September of 1996 and 1997. The differences in the hosts' habitats were reflected by the internal pools of DIC in these organisms. The concentrations of DIC in coelomic fluid from R. piscesae were 3.1 to 10.5 mM, lower than those previously reported for R. pachyptila, which often exceed 30 mM. When symbionts from both hosts were incubated at in situ pressures, their carbon fixation rates increased with the extracellular concentration of CO₂, and not HCO₃ ⁻, and symbionts from R. piscesae had a higher affinity for CO₂ than those from R. pachyptila (K _1/2 of 7.6 μM versus 49 μM). Transmission electron micrographs showed that symbionts from R. piscesae lack carboxysomes, irrespective of the coelomic fluid [DIC] of their host. This suggests that the higher affinity for CO₂ of R. piscesae symbionts may be their sole means of compensating for lower DIC concentrations. The δ¹³C values of tissues from R. piscesae with higher [DIC] in the coelomic fluid were more positive, opposite to the trend previously described for other autotrophs. Factors which may contribute to this trend are discussed. Received: 24 September 1998 / Accepted: 12 May 1999     

Release of phagocytosis-stimulating factor(s) by morula cells in a colonial ascidian

In vitro yeast phagocytosis by haemocytes of the compound ascidian Botryllus schlosseri was studied, with particular attention to interactions among different immunocyte types. It is demonstrated that the supernatant from haemocyte cultures matched with yeast cells contains factor(s) able to enhance yeast ingestion by Botryllus phagocytes. The increase in phagocytosis is not the consequence of yeast opsonisation, as the phagocytic index does not significantly increase when yeast cells, previously incubated in the culture media, are washed and re-suspended in filtered sea water. When haemocytes were fractionated by density gradient centrifugation and each band was incubated with yeast, the ability to stimulate phagocytosis was found in the supernatants from haemocyte cultures of fractions rich in morula cells (MC). Previous studies have demonstrated that MC express molecules recognised by anti-cytokine antibodies, as a consequence of the recognition of foreign molecules or cells. Our results indicate that molecules immunoreactive with anti-cytokine antibodies are required for modulating phagocyte activity, as the above-reported enhancing effect is completely absent in the presence of anti-IL-1-α and anti-TNF-α, but not of anti-rabbit-IgG antibodies, and they also highlight the presence of ‘cross-talk‘ between MC and phagocytes. A new scenario is therefore sketched, in which MC actively recognise non-self molecular patterns and, upon this recognition, release humoural factor(s) recognised by phagocytes, which modulate phagocytosis.     

Origin and development of the amoebocytes of Nicolea zostericola (Polychaeta: Terebellidae) with a discussion of their possible role in oogenesis

Three types of amoebocytes are present at different stages in the life history of Nicolea zostericola (Grube, 1860): a granular amoebocyte or spindle cell and 2 agranular amoebocytes, designated Type I and Type II. The agranular amoebocytes originate from specific sites on the lateral parietal peritoneum of the coelomic cavity in all the thoracic and a few abdominal segments. The origin of the granular amoebocyte is unknown. In juvenile worms the Type I amoebocyte enters the coelomic fluid and accumulates lipid and glycogen, presumably derived from reserves stored in the parietal peritoneum. Near the beginning of the vitellogenic phase of oogenesis, the Type I amoebocyte disappears and a Type II form is produced by the peritoneum. Type II cells differ cytologically from the Type I form by containing extensive rough ER (endoplasmic reticulum), Golgi complexes and free ribosomes; they appear to synthesize protein for export. At the end of vitellogenesis and near spawning, amoebocytes undergo alterations which include the migration of the nucleus to a central position, the formation of stacks and whirls of rough ER cisternae, and the appearance of vacuoles throughout the cytoplasm. It is suggested that the agranular amoebocytes obtain nutrients from food reserves stored in the parietal peritoneum and transfer them to the oocytes for use in yolk synthesis. The function of the granular amoebocytes is unknown.Contribution No.37 from the Marine Science Institute, Northeastern University, Nahant, Massachusetts, USA.Contribution No. 50, Harbor Branch Foundation, Inc.     

Genetic evidence of the presence of two species of Crassostrea (Bivalvia: Ostreidae) on the coast of Brazil

 Although oysters are commercially very important in Brazil, there is still much dispute about the number of Crassostrea species occurring on the Brazilian coast. The dispute is centered around C. brasiliana, considered by some authors to be a junior synonym of C. rhizophorae. In this paper we compared, by allozyme electrophoresis, sympatric and allopatric populations of the two putative species. Of the 17 loci analysed, five were diagnostic for the two species in sympatry (gene identity = 0.46 to 0.47), clearly demonstrating that they are distinct biological species. Heterozygosity (h) levels were high for both species (h = 0.24 to 0.28), and no heterozygote deficiencies were observed in any population (local inbreeding, F _IS  = 0.141; P > 0.70). Levels of population structure in C. rhizophorae along 1300 km of coast were very low (population inbreeding, F _ST  = 0.026; P > 0.15), indicating that the planktonic, planktotrophic larvae of these species are capable of long-range dispersal. Received: 14 January 1999 / Accepted: 8 December 1999     

Genetic differentiation in Hediste diversicolor (Polychaeta: Nereididae) for the North Sea and the Baltic Sea

Genetic differentiation between North Sea and Baltic Sea Hediste diversicolor (O.F. Müller, 1776) (Polychaeta: Nereididae) populations was studied by allozyme electrophoresis on starch gel. Thirteen loci were analyzed in eight populations. The level of genetic variation was very low (mean H _o = 0.000 to 0.015). Differentiation between H. diversicolor populations is quite high (F _ST = 0.892) and reflected by three enzyme loci (MDH-I*, MDH-II*, IDH-I*). The reduced gene flow (N _m<1) may be explained by the limited dispersal capacity of the species. Regardless of whether found in the North Sea or Baltic Sea, there appear to be two different genetic types which are parapatric or sympatric in some places. The two types hybridize at three localities, but no signs of hybridization have been found at one (Tallinn). Received: 26 June 1997 / Accepted: 10 September 1997     

Cysteine-glutathione disulfide, the sperm-release pheromone of the marine polychaete Nereis succinea (Annelida: Polychaeta)

Summary. In the marine polychaete Nereis succinea (Frey & Leuckart 1847) a sex pheromone was isolated from the coelomic fluid of sexually mature females and identified by NMR studies and independent synthesis. This pheromone is released by the females during reproduction together with eggs and coelomic fluid into the free water column and induces sperm release of surrounding males. Its structure was ascertained as L-cysteine-glutathione disulfide. It exhibited a response threshold of 0.6 · 10⁻⁷ M. Received 15 July 1997; accepted 25 November 1997.     

Restricted gene flow and evolutionary divergence between geographically separated populations of the Antarctic octopus Pareledone turqueti

Samples of the Antarctic octopus Pareledone turqueti were taken from three locations on the Scotia Ridge in the Southern Ocean. The genetic homogeneity of these populations was investigated using isozyme electrophoresis. Whilst panmixia appeared to be maintained around South Georgia (F _ST = 0) gene flow between this island and Shag Rocks, an island only 150 km away but separated by great depths, was extremely limited (F _ST = 0.74). These results are examined with respect to the discontinuous distribution of P. turqueti throughout Antarctica. An estimate of effective population size was also calculated (N _e = 3600). Received: 7 March 1997 / Accepted: 27 March 1997     

Genetic relationships between marine and marginal-marine populations of Cerithium species from the Mediterranean Sea

Results of isozyme electrophoresis were used to explore the genetic relationships between several Mediterranean morphs of Cerithium (Gastropoda: Prosobranchia), for which taxonomy is currently uncertain because of high intraspecific variability and low interspecific differentiation. The large species, classically known as C. vulgatum Bruguière, 1789 was identified at four sites (two in the French Mediterranean and two in southern Spain). Two different larval types were found in the French sites, but poecilogony could not be demonstrated. Individuals collected from harbours were not genetically distinct from open-sea populations of classic C. vulgatum. However, a population in the Embiez lagoon (French Mediterranean) which morphologically resembles C. vulgatum did display distinct genetic traits, supporting its status as a separate species. Of the small Cerithium species usually known as C. rupestre, two sympatric species (C.“rupestre” Risso, 1826 and C. lividulum Risso, 1826) were distinguished. Genotype frequencies within the analysed populations revealed much heterozygote deficiency. F _ST values (fixation index measuring the effects of population subdivision) suggest a higher genetic differentiation for C. lividulum populations than for C. vulgatum populations. We assume that a high larval dispersal capability (via planktotrophy) allows a high gene flow between populations of C. vulgatum. Received: 24 November 1998 / Accepted: 24 September 1999     

Role of protein in larval swimming and metamorphosis of Bugula neritina (Bryozoa: Cheilostomatida)

Larvae of the marine cheilostomatid bryozoan Bugula neritina (L.) were prevented from settling for 1, 4 and 8 h by mechanical agitation, following which settlement and metamorphosis success were examined. Settlement rates were significantly affected by swimming time, which decreased from 100% after 2 h to 93.7 ± 4.3% after 8 h. Similarly, metamorphosis to the feeding ancestrula was significantly impaired following a swimming time of 8 h, declining from 93.7 ± 4.3% after 1 h to 65.9 ± 7.0% after 8 h. The resultant colonies grew well for the first 3 wk, following which time, growth patterns became erratic. Growth rate was in all cases highly variable, and did not correlate with enforced swimming times. Larval protein composition was examined after 1, 4 and 8 h swimming time, and post-larval composition 1, 2, 5, 24 and 48 h after settlement using sodium-dodecyl-sulphate polyacrylamide-gel electrophoresis (SDS-PAGE). Individual protein content was measured using a densitometer. Larvae did not consume protein during swimming, however a protein measuring 170 kdaltons was consumed during metamorphosis. These results are discussed in the context of larval settlement and energetics. Received: 19 July 1998 / Accepted: 3 December 1998     

Genetic divergence and phylogenetic inferences in five species of Mugilidae (Pisces: Perciformes)

Allozyme electrophoresis was used to compare the genetic divergence of global populations of Mugil cephalus Linnaeus, 1758 and two congeneric [M.␣curema Cuvier & Valenciennes, 1836; M. gyrans (Jordan & Gilbert, 1885)] and two more distantly related [Liza ramada (Risso, 1826); Xenomugil thoburni (Jordan & Starks, 1896)] species on the basis of 18 enzyme loci. The amount of genetic divergence among the species examined is in agreement with their present systematic status, the differences being larger among non-congeneric species than among species of the same genus. Intraspecific genetic distances in M. cephalus (average Nei's D= 0.154), although higher than those reported for conspecific populations of fish, appear to be small when compared to the interspecific values among mugilid species (0.821 ≤ Nei's D ≤ 1.744). Phylogenetic trees obtained by genetic distance methods and discrete character parsimony analysis were of similar topology, except for the relationships within the genus Mugil and for the arrangement of M. cephalus populations. Received: 7 April 1997 / Accepted: 4 February 1998     

Damage to grass shrimp (Palaemonetes pugio) embryo DNA by summer sunlight followed by DNA repair in the dark

 DNA strand damage, using the single-cell gel electrophoresis (comet) method, was determined in different-stage embryos of grass shrimp (Palaemonetes pugio) collected from surface waters of a local estuary near Savannah, Georgia, USA. Late-stage embryos collected from the estuarine river at midday in the summer or placed in a solar simulator showed extensive DNA strand damage. The solar simulator, which produced the total irradiance found at midday in the summer at 34°N caused DNA strand damage in embryos similar to that found in sunlight-exposed embryos. A large increase in cyclobutane pyrimidine dimers (18 cyclobutane pyrimidine dimers per 1000 kilobases) were detected in DNA from the late-stage embryos exposed to the solar simulator for 1 h (solar irradiance of 126 μW cm⁻²). DNA repair took place within a few hours when late-stage embryos collected at midday from the river were transferred to the dark. Early grass-shrimp embryo stages showed no DNA strand damage either after placement in the solar simulator or when collected at midday in the summer. This lack of solar-damaged DNA in early-stage embryos was probably due to the presence of high concentrations of carotenoids, which can act as anti-oxidants to prevent damage from activated oxygen species produced by cells exposed to ultraviolet light. These carotenoids are utilized by the developing embryos, and only low concentrations of carotenoids were present in late embryo stages. Received: 24 May 1999 / Accepted: 11 May 2000     

Genetic variation and clonal structure in the scleractinian coral Pocillopora damicornis in the Ryukyu Archipelago, southern Japan

Samples of the scleractinian coral Pocillopora damicornis were collected from six sites located around four islands in the Ryukyu Archipelago, southern Japan, and subjected to allozyme electrophoresis. Seven polymorphic loci were examined for their allelic patterns. The ratio of observed to expected genotypic diversity (0.30 < G _o :G _e  < 0.64), the ratio of the observed number of genotypes to the number of individuals (0.47 < N _g :N _i  < 0.75), and deviations from Hardy–Weinberg equilibrium indicated that asexual reproduction plays a major role in the maintenance of established populations. However, populations were not completely dominated by a single or a few clones, and most clones were represented by only a few individual samples. The high frequency of typhoons in the region suggests that, in P. damicornis, fragmentation caused through occasional exposure to powerful waves is a major mode of asexual reproduction, but asexual production of planulae may also be contributing to the maintenance of populations. A significant genetic differentiation (F ST) was found between the six populations examined (0.027 < F ST < 0.092, average F ST = 0.056). The moderate gene flow is discussed according to characteristics of the larval stage of the species, and to circulation patterns in the region. Received: 7 August 1998 / Accepted: 18 May 1999     

Sexuality and cyst formation of the spring-bloom dinoflagellate Scrippsiella hangoei in the coastal northern Baltic Sea

The temporal sequence and the magnitude of the sexual reproduction and subsequent cyst deposition of the common spring-bloom dinoflagellate Scrippsiella hangoei (Schiller) Larsen was studied during spring 1996 on the SW coast of Finland, Baltic Sea. The abundances of the different size of fractions of S. hangoei (14 to 18 μm, 18 to 22 μm and >22 μm) were monitored in the water column, and the deposition of resting cysts was measured using moored sediment traps. Cyst sedimentation rates were measured throughout the seasonal cycle in order to estimate cyst resuspension rates for the quantitative assessment of the fraction of population undergoing encystment. The onset of sexual reproduction, indicated by a significant increase of the small cells (14 to 18 μm) representing gametes, occurred in a nutrient replete environment well before the exponential growth phase and peak abundances of vegetative cells. Gamete formation was followed by high abundances of large cells (>22 μm) representing planozygotes, and subsequent sedimentation of resting cysts. Approximately 60% of the asexually growing bloom population was estimated to form planozygotes, suggesting that encystment was an important factor in bloom termination and possibly plays a role in the regulation of the magnitude of the bloom. Finally encystment accounted for 40% of the entire S. hangoei population, resulting in a considerable loss of the bloom population and an input of the vernal phytoplankton biomass to the benthos. Received: 11 December 1998 / Accepted: 8 April 1999     

Population genetics of the Antarctic heteronemertean Parbolasia corrugatus from the South Orkney Islands

An allozyme survey, using starch-gel electrophoresis, was carried out on eight populations of the Antarctic nemertean worm Parborlasia corrugatus (McIntosh, 1876) collected from locations around the South Orkney Islands, Antarctica. These populations were separated by distances in the order of tens of kilometres. Genetic variation was estimated over 22 enzyme loci for all populations examined, giving an observed heterozygosity of 0.142. This was much lower than the expected heterozygosity (H _e  = 0.201), and it was found that there was a significant deficiency of het‐erozygotes across four enzyme loci ( p ≤ 0.01). A more detailed examination of this deficiency of heterozygotes was undertaken for the six populations and six variable enzyme loci for which the most complete data sets existed. A significant deficiency of heterozygotes was found at the enzyme locus Odh-1 for four of the six populations examined ( p ≤ 0.01). Mean F _is (0.240) indicated a significant ( p ≤ 0.01) within-population component of the heterozygote deficiency estimated for the six populations sampled, and this was mainly due to the␣Ap-1, Odh-1 and Pgm-1 loci. The mean F _st value (0.036) was also significant ( p ≤ 0.01), indicating a degree of genetic differentiation between populations. The observed levels of genetic differentiation between populations of P. corrugatus and the significant heterozygote deficiencies were unexpected, because this species has been reported to have a long-lived planktotrophic larva. It is hypothesised that recruitment of P. corrugatus in the South Orkney Islands originates from genetically distinct populations located in the Weddell Sea and to the west of the Antarctic Peninsula. Shifts in the relative position of the Weddell Sea Front, Weddell–Scotia Confluence and Scotia Front, relative to the South Orkney Islands, provide a mechanism for variation in the origin of recruits over time. Received: 24 July 1997 / Accepted: 31 October 1997     

Is Syllis gracilis (Polychaeta: Syllidae) a species complex? An allozyme perspective

The genetic relationships between morphologically indistinguishable marine and brackish populations of Syllis gracilis Grube, 1840 (Polychaeta: Syllidae) were studied by means of allozyme electrophoresis. Samples of S. gracilis from marine coastal and brackish-water habitats were examined for variation at 13 presumptive loci. In addition, a sample of the closely related species S. prolifera (Krohn, 1852) was analysed. Five loci were multiallelic in at least one population of S. gracilis and eight loci in S. prolifera. Low to moderate levels of within-population genetic variability were found, with average expected heterozygosity values ranging from H = 0.068 (±0.043 SE) to 0.187 (±0.069 SE) in the populations of S. gracilis; higher values were found in S. prolifera (H = 0.325 ± 0.076). The presence of various private alleles indicated a marked genetic divergence among populations of S. gracilis, with Nei's genetic distances ranging from D = 0.000 to 0.833 and a highly significant F _ST value. Furthermore, evidence for strong genetic heterogeneity between two sympatric marine populations was found. UPGMA cluster analysis and multidimensional scaling pointed out a clear genetic divergence between brackish and marine populations. At least two genetically divergent entities occurred in marine and brackish habitats. This could be due to local adaptation of individuals coming from marine populations to brackish habitats, but more presumably to the occurrence of a species complex within S. gracilis. Received: 6 June 1999 / Accepted: 7 February 2000     

Effects of host-tissue homogenate of the scleractinian coral Plesiastrea versipora on glycerol metabolism in isolated symbiotic dinoflagellates

Symbiotic dinoflagellate algae (Symbiodinium sp.) isolated from the scleractinian coral Plesiastrea versipora and incubated in homogenized host tissue released 4 to 7 times as much glycerol (14 to 46 nmol glycerol/10⁶ algae) as those incubated in seawater (3 to 6 nmol glycerol/10⁶ algae) after 4 h incubation in the light. During this period, no release of triglycerides was detected. Intracellular glycerol increased 2- to 3-fold in algae incubated in host homogenate, but remained unchanged in algae incubated in seawater at a concentration of 0.82 ± 0.47 nmol glycerol/10⁶ algae. In each incubation condition, intracellular triglyceride levels increased. However, in algae incubated in host homogenate, the intracellular levels of triglycerides reached only about 75% of the amount reached in algae incubated in seawater (max. 18.55 ± 2.40 nmol glycerol/10⁶ cells). Host homogenate did not stimulate the release of glycerol from algae during dark incubation. These data show that the glycerol released by algae incubated in host-tissue homogenate was derived from increased synthesis of glycerol or from diversion of some glycerol or other photosynthetic intermediates from incorporation into algal triglyceride stores, and did not come from existing stores. Received: 20 December 1996 / Accepted: 9 January 1997     

Resting spore formation and biochemical composition of the marine planktonic diatom Chaetoceros pseudocurvisetus in culture: ecological significance of decreased nucleotide content and activation of the xanthophyll cycle by resting spore formation

The biochemical composition of vegetative cells and resting spores of Chaetoceros pseudocurvisetus Mangin was compared in cultures under various nutrient and light conditions. The cellular content of major nucleotides such as AMP, ADP, ATP and UTP decreased in the order: vegetative cells, nutrient-starved (vegetative) cells and resting spores, indicating that the general metabolism of resting spores is relatively inactive. ADP-glucose was only abundant in nutrient-starved vegetative cells, suggesting metabolic imbalance in these cells. The chl a–specific fluorescence yield of vegetative cells grown under all culture conditions was low, but very high in resting spores. The ratios of the cellular contents of diadinoxanthin to chl a and of diatoxanthin to chl a were higher in resting spores and nutrient-starved vegetative cells than in nutrient-replete vegetative cells. The diadinoxanthin–diatoxanthin xanthophyll cycle was active in resting spores; the xanthophyll cycle was synchronized with a 14 h light:10 h dark photoperiod. Also, the ratios of cellular content of diadinoxanthin and diatoxanthin to cellular content of chl a in resting spores were relatively high in high irradiance, and decreased gradually in conditions of darkness over long culture periods. Under conditions of strong light and high temperature, most resting spores survived more than 40 d while nutrient-starved vegetative cells died within 33 d. These results suggest that resting spore formation is a strategy for enhancing protection and lowering metabolic rate for survival. These physiological changes accompanying spore formation enable resting spores not only to overwinter but also to “oversummer” in the coastal euphotic layer. Received: 23 March 1999 / Accepted: 11 August 1999     

Role of adipocytes in the muscle tissue of Atlantic salmon (Salmo salar ) in the uptake, release and retention of water-soluble fraction of crude oil hydrocarbons

The uptake and depuration of the water-sol‐uble fraction (WSF) of hydrocarbons of crude petroleum by Atlantic salmon (Salmosalar) has previously been examined in terms of whole muscle. The hypothesis that the tainting WSF in the muscle was retained primarily by adipocytes has been investigated by the isolation of adipocytes and the subsequent analysis for hydrocarbons in adipocytes. After 96 h exposure of market-sized Atlantic salmon to 0.2 ppm WSF, adipocytes isolated from the belly flap region of the muscle tissue accumulated 14.3 times more WSF (59.4 ppm) than the dorsal white muscle (4.2 ppm), while 54% of the tainting WSF in the dorsal white muscle was found to be stored in associated adipocytes. When returned to clean seawater, WSF accumulated in the dorsal white muscle was released much faster than that in the adipocytes. These results indicated that the loose association of WSF with the nonlipid portion of white muscle, mainly muscle cells and intercellular fluid, is responsible for the rapid discharge of WSF from the dorsal muscle tissue in the early stages of depuration. After 4 d of depuration, the adipocytes became the principal storage site of residual WSF in white muscle and the depuration of WSF from muscle tissue then reflected the release of WSF from adipocytes in the muscle tissue. After 20 d of depuration, 10.7 ppm of tainting WSF in the form of high molecular weight aromatic hydrocarbons (mainly C₄-benzenes, naphthalene and alkylated naphthalenes) were still present in adipocytes, while in the dorsal white muscle only a trace of total WSF was detected. Increases in the number of aromatic rings and the alkylations on the rings enhanced the accumulation and retention of individual hydrocarbons in both adipocytes and white muscle. From these studies we conclude that it is the adipocytes in the muscle tissue which control the actual accumulation and release of hydrocarbons in the whole muscle tissue of Atlantic salmon. Received: 21 August 1996 / Accepted: 26 September 1996     

Effects of organochlorines on the ultrastructure of the liver of the damselfish Parma microlepis from reefs in New South Wales, Australia

  Parma microlepis (Günther) were collected from Malabar, an urban location close to the centre of Sydney, Australia, and from Jervis Bay, a reference location 170 km south of the city centre. At each location, fish were collected from two sites separated by 100 to 200 m. The ultrastructure of normal liver tissue is described based on 20 female fish collected from Jervis Bay, where fish are known to be exposed to low levels of organochlorine contaminants. Alterations in the endoplasmic reticulum, mitochondria, lysosomes and nuclei of hepatocytes were identified and quantified in the liver tissue of fish from this location and compared to alterations in 20 female fish collected from Malabar, where fish are exposed to higher concentrations of organochlorine pesticides such as DDT compounds. There were significant differences in the percentage of hepatocytes with swollen mitochondria (F = 124.025, df = 2, 2, P = 0.008) and atypical nuclei (F = 22.198, df = 2, 2, P = 0.043) between sites (100 to 200 m apart), but there were no clear differences between the percentage of structural alterations in the hepatocytes of P. microlepis from Jervis Bay and Malabar. Associations between liver morphology and the organochlorines aldrin, dieldrin, DDE and chlordane were examined using a Pearson correlation matrix. Significant correlations were detected between the percentage of hepatocytes with dilated endoplasmic reticulum and the concentrations of the pesticide aldrin (r = 0.600, df = 11, r _{crit(α = 0.05)} = 0.553). Significant associations were also detected between the percentage of hepatocytes with disorganised endoplasmic reticulum and the concentrations of dieldrin and DDE residues in fish (r = 0.576, r = 0.567, respectively, df = 13, r _{crit (α = 0.05)} = 0.514). However, there was little evidence that ultrastructural alterations in fish responded to increasing concentrations of these pesticides in a consistent dose-response manner. Received: 20 October 1998 / Accepted: 24 November 1999     

Importance of cysts in the population dynamics of the red tide flagellate Heterosigma akashiwo (Raphidophyceae)

To elucidate roles of cysts in occurrences of Heterosigma akashiwo blooms, cyst dynamics were studied in northern Hiroshima Bay, the Seto Inland Sea of Japan, where H. akashiwo regularly forms red tide in June. Monthly measurements of seasonal changes in the densities of vegetative cells of H. akashiwo and their germinable cysts in surface sediments (top 1-cm layer) were made for 2 years at three stations. Vegetative cells of H. akashiwo could be detected from April through December throughout the water column, and the existence of vegetative cells was confirmed in surface waters even in winter after incubation of sampled seawater in culture medium. Germinable cysts, enumerated by the extinction dilution method, existed in sediments in all seasons, even before and after the seasonal bloom. The effects of incubation temperature on the germination of natural cysts of H. akashiwo in sediments were examined. Germination was not observed at 5 °C, was low at 10 °C, while it increased at 15 °C, and maintained a high level to 25 °C. The bottom water temperature reached 15 °C (suitable for the germination of cysts) and the surface about 18 °C or more (suitable for the growth of vegetative cells) 2 to 3 weeks before the blooms. The dark survival of H. akashiwo cysts was tested, and it was found that the cysts were viable for at least 650 d at 11 °C, and for 165 d at 25 °C, indicating a significant role of cysts in the survival during winter and summer seasons. The cysts presumably also play an important role in seeding primary populations into water columns when the bottom water reaches a suitable temperature (around 15 °C); thereafter the populations develop with great annual regularity to bloom in June. These results suggest that initiation of H. akashiwo red tides in the Seto Inland Sea could be triggered by bottom water temperature. Received: 3 July 1998 / Accepted: 12 January 1999