RAPD-PCR investigations of systematic relationships among four species of eels (Teleostei: Anguillidae), particularly Anguilla anguilla and A. rostrata

 We examined the genetic identities of 49 individuals of four species of eels in the genus Anguilla Shaw, A. anguilla, A. japonica, A. reinhardti and A. rostrata, using the random(ly) amplified polymorphic DNA (RAPD)-PCR technique. We used 15 random decamer primers. Out of the 22 148 amplification products detected, 454 markers were evaluated using Nei's distance coefficient, two numerical methods (neighbor-joining, UPGMA), and phylogenetic analysis using parsimony (PAUP). Both numerical and parsimony methods were congruent and confirmed the specific status of all four examined Anguilla species. In all dendrograms, all individuals of each of the species clustered together within four closed groups with highly significant bootstrap values (between 97 and 100%). Additionally 1 to 18 species-specific monomorphic diagnostic fragments were detected in three of the species; none were detected in A. anguilla. Mean interspecific genetic distances ranged from 0.384 to 0.559, with the minimum between A. anguilla and A. rostrata and the maximum between A. anguilla and A. reinhardti. Individuals collected at one locality and belonging to a single species did not cluster together, and neither were any diagnostic monomorphic fragments found for individuals of single localities. This probably reflects a random dispersal of larvae in ocean currents before they reach coastal waters. Among all possible species pairs, only the two Atlantic species were consistently clustered in all dendrograms, with highly significant bootstrap values (100%). Additionally, we detected ten diagnostic markers for this pair of eel species. From RAPD data, we suggest a phylogram which was routed by the descending analysis method: A. reinhardti appears to have been the first species to diverge from a putative common ancestor of the four eel species. Later, A. japonica separated from the hypothetical ancestor of the Atlantic species, which then split recently into A. anguilla and A. rostrata. The close relationship of A. anguilla and A. rostrata is indicated by their low genetic distance (GD = 0.384) and a high degree of shared RAPD fragments (SF = 71.2%); this compares with the following means for all other species pairs: ( = 0.531,  = 44.2%). The prerequisites for using RAPD data for the reconstruction of phylogenies are discussed. Received: 2 September 1999 / Accepted: 8 May 2000     

Parthenogenetic reproduction of Diaphanosoma celebensis (Crustacea: Cladocera): influence of salinity on feeding, survival, growth and neonate production

 Effect of salinity on the feeding rate and parthenogenetic reproduction of asexual females of the cladoceran Diaphanosoma celebensis Stingelin was studied. Short-term (10 h) grazing experiments were conducted using Isochrysis galbana as feed at 5, 17, 25 and 30 psu salinity. Gut pigment concentration showed a significantly higher rate of feeding at lower salinities. Survival, growth, maturity attainment and neonate production of asexual females reared in the above four test salinities indicated preference for lower salinities (5 and 17 psu). The mean size of adult females decreased from 909 to 593 μm, mean life span from 24 to 5 d, mean neonate production from 12 to 2 and mean size of neonates from 434 to 400 μm as the salinity increased from 5 to 30 psu. Salinity variations also affected the size and age of primiparous females. Resting egg formation and sexual reproduction did not occur at the tested salinities. The results indicate that D. celebensis is adapted to low saline, estuarine environments. Received: 14 January 2000 / Accepted: 24 March 2000     

Genetic differentiation in a pelagic crustacean (Meganyctiphanes norvegica: Euphausiacea) from the North East Atlantic and the Mediterranean Sea

Meganyctiphanes norvegica (M. Sars)is a pelagic crustacean that plays a key role in marine food webs of North Atlantic Ocean and marginal seas. We studied eight population samples collected in the European Atlantic and Mediterranean Sea. By means of single strand conformation polymorphism analysis (SSCP) and direct sequencing, we investigated a segment of 158 base pairs of the mitochondrial gene coding for the subunit 1 of NADH dehydrogenase. We found 12 sequence variants among the 385 individuals studied. Analysis of molecular variance (AMOVA) showed that 14.75% of the total genetic variability was explained by differences between populations, thus indicating absence of panmixia for these populations. Pairwise comparisons revealed three distinct genetic pools: the first one represented by Cadiz Bay, the second one by the Ligurian Sea, and the third one included all the NE Atlantic samples. We also investigated one population from the Alboran Sea (within the Mediterranean basin, east of the Strait of Gibraltar). This population was found to be genetically intermediate between the NE Atlantic samples and the Ligurian sample, suggesting that the restriction to the gene flow is not associated with the Strait of Gibraltar, but possibly with the Oran–Almeria oceanographic front. The present work indicates that M. norvegica, although endowed with a high dispersal capacity because of its pelagic habit, can develop separate breeding units inside the same oceanic basin (the Atlantic). Furthermore, the Ligurian sample should be considered as a distinct evolutionary entity, separated from the Atlantic population. Received: 2 May 1999 / Accepted: 26 November 1999     

Effects of dissolved ammonium addition and host feeding with Artemia salina on photoacclimation of the hermatypic coral Stylophora pistillata

 Effects of nutrient treatments on photoacclimation of the hermatypic coral Stylophora pistillata (Esper) were studied. Studies on photoacclimation of colonies from different light regimes in the field were evaluated and used to design laboratory experiments. Coral colonies were collected in the Gulf of Eilat (Israel) from January to March 1993. Exterior branches of colonies from different depths (1 to 40 m) displayed different trends in production characteristics at reduced and very low levels of illumination. From 24 ± 3% to 12 ± 2% of incident surface photosynthetic active radiation (PAR_o), zooxanthella population density and chlorophyll a+c per 10⁶ zooxanthellae increased, a trend seen in the range of light levels optimal for coral growth (90 to 30% PAR_o). The P _max of CO₂ per 10⁶ zooxanthellae decreased, while P _max of CO₂ per 10³ polyps increased, indicating an increase in zooxanthella population density at low light levels. Proliferous zooxanthella frequency (PZF, a measure of zooxanthella division) declined significantly at light levels <18 ± 3% PAR_o. At the lowest levels of illumination (<5% PAR_o), zooxanthella population density decreased, as did the PZF; chl a+c per 10⁶ zooxanthellae was unchanged. In 28-d experiments, exterior coral branches from the upper surfaces of colonies from 3 m depth (65 ± 4% PAR_o) were incubated in aquaria under bright (80 to 90% PAR_o), reduced (20 to 30% PAR_o), and extremely low (2 to 4% PAR_o) light intensities. At each light intensity, the corals were maintained in three feeding treatments: sea water (SW); ammonium enriched SW (SW + N); SW with Artemia salina nauplii (SW + A). An increase in P _max of CO₂ per 10³ polyps was found in corals acclimated to reduced light (20 to 30% PAR_o) in nutrient-enriched SW, while in SW, where the increase in zooxanthella population density was smaller, it did not occur. Nutrient enrichments (SW + N at 2 to 4% PAR_o and SW + A at 20 to 30% PAR_o) increased zooxanthella population density, but had no effect on chl a+c per 10⁶ zooxanthellae. Acclimation for 14 d to reduced (10 to 20% PAR_o) and extremely low (1 to 3% PAR_o) light intensities shifted ¹⁴C photoassimilation into glycerol and other compounds (probably glycerides), rather than sugars. Both ammonium addition and feeding with Artemia salina nauplii resulted in an increase in photosynthetic assimilation of ¹⁴C into amino acids. We conclude that acclimation to reduced light consists of two processes: an increase in photosynthetic pigments and in zooxanthella population density. Both processes require nitrogen, the increase in zooxanthella population density needing more; this adaptation is therefore limited in nitrogen-poor sea water. Received: 19 June 1998 / Accepted: 13 June 2000     

Metabolic rate and lipofuscin accumulation in juvenile European lobster (Homarus gammarus ) in relation to simulated seasonal changes in temperature

 The accumulation of lipofuscin, which is an indictor of physiological age, in the brain of juvenile European lobster (Homarus gammarus L.) was monitored for 22 mo in three experimental temperature regimes that simulated seasonal variation in temperature in the geographic range of this species. Metabolic rate responses to changes in temperature were estimated by measuring the activity of the electron transport system (ETS) in muscle tissue and in vivo rates of oxygen consumption. Lipofuscin accumulation oscillated with simulated seasonal changes in temperature and was described by seasonalised von Bertalanffy growth functions. The incremental accumulation in lipofuscin between sampling dates was linearly related to the number of degree days that accumulated between dates, irrespective of the amplitude of temperature fluctuation that had occurred. ETS activity increased with acclimation temperature and was modelled using a polynomial function. This indicated a lower temperature sensitivity in the temperature mid-range (12 to 16 °C), although the Q₁₀ for this mid-range was 2.1. ETS activity in lobsters acclimated to 8 and 18 °C and assayed at 13 °C was similar, indicating no compensation for changes in environmental temperature. Oxygen consumption rate was significantly higher at 14 °C than at 10.5 °C and had a Q₁₀ of 3.6, again suggesting no compensation to temperature change. The absence of metabolic compensation in response to temperature change in H. gammarus is consistent with the predictability of changes in temperature and food availability in the sub-littoral environment of this species. As lipofuscin accumulates according to metabolic rate, and metabolic rate in H. gammarus is directly correlated with temperature, geographic differences and long-term temporal trends in temperature will need to be considered when converting physiological age indices, obtained from lipofuscin estimates, to a chronological scale. Received: 27 April 2000 / Accepted: 21 July 2000     

Interannual, seasonal, local and body size variations in reproduction of the prawn Penaeus (Marsupenaeus) japonicus (Crustacea: Decapoda: Penaeidae) in the Ariake Sea and Tachibana Bay, Japan

 The reproductive biology of female Penaeus japonicus Bate was investigated in the Ariake Sea and Tachibana Bay (located outside the Ariake Sea), Japan from 1994 to 1996. Interannual, seasonal, individual female body size and spatial influences on the incidence of spawning were examined. The proportion of inseminated females, increased with increasing body size up to 170 mm body length (BL) and decreased thereafter. The minimum size at maturity was similar between years; 130 to 140 mm BL. The minimum size of individuals with developing ovaries or spermatophores differed from that of ripe females by 20 to 25 mm. Spawning occurred in the central part of the Ariake Sea and Tachibana Bay but rarely in the inner part. P. japonicus had a clear reproductive cycle. Spawning started earlier and ended later, occurring from mid-May to mid-October, in the eastern central part of and outside of the Ariake Sea compared with the western central part, where spawning occurred from mid-June to mid-September. Spawning dynamics differed across seasons, body sizes and areas but not across years. The seasonal peak in the proportion of ripe females varied with body size. In small individuals (130 to 169 mm BL), no peak was observed, whereas in large individuals (>170 mm BL) the proportion of ripe individuals peaked in June. The proportion of ripe individuals increased with increasing body size and was high outside the Ariake Sea. Factors causing the variation in spawning dynamics are discussed. Received: 30 November 1998 / Accepted: 12 October 1999     

Reproductive variability over a four-year period in the sea urchin Evechinus chloroticus (Echinoidea: Echinodermata) from differing habitats in New Zealand

 The endemic New Zealand echinoid, Evechinus chloroticus (Valenciennes), was sampled approximately monthly from September 1990 to October 1994 at three sites in Tory Channel, Marlborough Sounds, New Zealand. These channel sites (outer, mid and inner) were up to 20 km from the open ocean and differed in their shore type, exposure to wave action and macrophyte abundance. E. chloroticus showed an annual reproductive cycle at each site, with gametogenesis commencing in the late austral winter and spawning in summer. Maximum gonad indices (reproductive potential) varied spatially, with the outer site generally having higher maximum gonad indices than the inner site, and temporally, with maximum gonad indices occurring earlier in 3 of the 4 seasons at the outer site than the inner site. The mid and inner sites showed much greater variation in maximum gonad indices (range 15.83 to 26.99% and 11.87 to 20.90%, respectively) than the outer site (range 19.31 to 22.95%). Reproductive output (weight of gametes released per gram echinoid) also varied, with the different sites showing significantly different outputs in the different years. A regression of maximum reproductive potential against reproductive output was significant ( p < 0.001), and had a positive slope with an r ² of 0.79. While, the initiation of gametogenesis was relatively synchronous between sites and years, and is possibly cued by increasing daylength, it progressed at different rates among populations. Spawning did show spatial and temporal variability, occurring near the time of highest sea-surface temperatures (∼15 °C). The observed variations in reproductive cycle may be related to small-scale variability in diet and environmental conditions. Furthermore, asynchronous spawning, variable spawning duration, and variable reproductive output are likely to strongly influence annual recruitment variability in E. chloroticus, with different larval subpopulations contributing unequally in different years. The ecological consequences of this, both for the ability of E. chloroticus to propagate itself in space and time and for the management of the developing E. chloroticus fishery in New Zealand, are discussed. Received: 9 December 1998 / Accepted: 13 June 2000     

Predation on meiofaunal and macrofaunal invertebrates by western sandpipers (Calidris mauri): evidence for dual foraging modes

Western sandpiper (Calidris mauri) predation was examined by concurrent experiments and direct observations of foraging behaviour on high intertidal mudflats of the Fraser River estuary, British Columbia. Western sandpipers foraged by either “pecking” on the surface (64% of observational time) or probing into sediment (29%). The first experiment (probe-mark method) consisted of collecting small-volume cores (21.2 cm³) of probed (experimental) and non-probed (control) sediment on the tidal flat, following a 22.5-min feeding period. The second experiment (exclosure method) involved deploying exclosures immediately prior to the feeding period and subsequent collection of cores from inside (control) and outside (experimental) the exclosures. Sediment cores were analysed for both macrofaunal and meiofaunal size fractions. Comparisons between macro- and meiofaunal invertebrate densities in experimental and control sediments revealed significant differences, attributed to shorebird predation, for both experiments. The probe-mark experiment detected the removal of large infaunal polychaetes (∼ 20 mm), while the exclosure experiment showed depletion of epifaunal harpacticoid copepods (0.063–0.5 mm). Predation on macrofaunal cumaceans was detected in both experiments. Invertebrates selected by western sandpipers neither fell within traditional infaunal size classifications (macro- vs. meiofauna; 500 μm delineation) nor corresponded to the highest densities of taxa. Rather, inference from experimental results and observations is that western sandpipers forage in two modes, by: (1) surface gleaning of epibenthic copepods and cumaceans in the macro- and meiofaunal size ranges and (2) selective probing for larger infauna, such as polychaetes. These findings were facilitated by the combination of methodologies employed. Received: 29 December 1999 / Accepted: 11 September 2000     

Biomass partitioning of benthic microbes in a Baltic inlet: relationships between bacteria, algae, heterotrophic flagellates and ciliates

 The structure of a benthic microbial food web and its seasonal changes were studied in the shallow brackish waters of the island of Hiddensee, northeastern Germany, at two sites in close proximity by monthly or bimonthly sampling from July 1995 to June 1996. Abundance and biomass of phototrophic and non-phototrophic bacteria, heterotrophic flagellates (HF) and ciliates as well as the biomass of microphytobenthos were determined in the upper 0.3 cm sediment layer. Abundance of organisms showed strong positive correlation with water temperature, with the exception of the bacteria. Non-phototrophic bacterial numbers ranged from 7 × 10⁸ to 6.7 × 10⁹ cells cm⁻³ and phototrophic bacterial abundance from 4 × 10⁷ to 2.7 × 10⁸. Heterotrophic protist abundance ranged from 8 × 10³ to 104 × 10³ ind cm⁻³ for HF and from 39 to 747 ind cm⁻³ for ciliates. The biomass partitioning demonstrated the primary importance of non-phototrophic bacteria (min. 0.83, max. 84.87 μg C cm⁻³), followed by the microphytobenthos (min. 1.32, max. 50.93 μg C cm⁻³). The heterotrophic protists contributed roughly the same fraction to the total microbial biomass, with the biomass of the HF being slightly higher (HF 0.23 to 1.76 μg C cm⁻³, ciliates 0.04 to 1.17 μg C cm⁻³). Taxonomic classification of the benthic HF revealed the euglenids to be the most important group in terms of abundance and biomass, followed by thaumatomastigids and kinetoplastids. Other important groups were apusomonads, cercomonads, pedinellids and cryptomonads. The structure of the HF assemblage showed strong seasonal changes with euglenids being the most abundant taxa in summer, while apusomonads and thaumatomastigids were predominant in winter. Similar to the pelagic microbial food web, benthic picophototrophic bacteria were occasionally abundant, and the feeding modes of heterotrophic protists exhibited a great variety (predominantly omnivores, bacterivores, herbivores or predators). Filter-feeding HF were of little importance. Contrary to the pelagic environment, a top-down control on total benthic bacterial numbers by HF seemed unlikely at the studied stations which were characterised by muddy sand. Received: 6 January 1999 / Accepted: 21 October 1999     

Grazing rates of organic matter and living fungal biomass of decaying Spartina alterniflora by three species of salt-marsh invertebrates

 The pathway for the flow of salt-marsh grass production into marsh food-webs is still not well defined. We compared the abilities of three marsh macroinvertebrates [salt marsh periwinkles, Littoraria irrorata (Say) (=Littorina irrorata), salt-marsh coffee-bean snails, Melampus bidentatus (Say); and a talitrid amphipod, Uhlorchestia spartinophila Bounsfield and Heard] to access standing-dead leaves of smooth cordgrass (Spartina alterniflora Loisel). The invertebrates were incubated with naturally-decaying leaves, and the rates of removal of organic matter and living fungal biomass (ergosterol) were measured. The impact of invertebrate activity upon fungal growth rates was measured as rates of fungal-membrane synthesis (incorporation of radioacetate into ergosterol). The removal rates of organic leaf biomass per mg individual biomass were highest for amphipods (700 μg mg⁻¹ d⁻¹) and lowest for periwinkles (90 μg mg⁻¹ d⁻¹), but the relatively large biomass of the snails made their removal rates per individual greater than those of amphipods. Net removal of ergosterol by all three invertebrates was >50% for yellow-brown (early-decay) leaf blades. For fully-brown (advanced-decay) blades, >50% removal of ergosterol was found only for periwinkles; exposure to coffee-bean snails and amphipods resulted in a net ergosterol reduction of ≤20%. The lower net reduction of living fungal biomass by coffee-bean snails and amphipods may have been due to fungal-growth stimulation (2.3-fold stimulation in coffee-bean snails and 1.5-fold stimulation in amphipods). Grazing by periwinkles did not stimulate fungal growth, possibly because of its high intensity. Grazing by these three salt-marsh shredders may affect marsh-grass shoot-decay in different ways. Periwinkles may abbreviate the period of fungal production, and incorporate the decaying material relatively quickly into snail biomass and fecal-pellet rain to the sediments. Coffee-bean snails and amphipods may enhance and prolong fungal production, along with the formation of fecal-pellet rain. All three invertebrates fed preferentially on leaf blades rather than leaf sheaths, and feeding rates of gastropods were higher during the night than during the day. Received: 25 November 1998 / Accepted: 4 November 1999