Respiration of the eggs of the giant cuttlefish Sepia apama

On the roofs of subtidal crevices, the giant cuttlefish (Sepia apama) of southern Australia lays clutches of lemon-shaped eggs which hatch after 3 to 5 mo. Diffusion of oxygen through the capsule and chorion membrane to the perivitelline fluid and embryo was modelled using the equation V˙ _O2 = G _O2(P _O2out−P _O2in), where V˙ _O2 = rate of oxygen consumption, G _O2 = oxygen conductance of the capsule, and P _O2 values = oxygen partial pressures across the capsule. During development, V˙ _O2 rose exponentially as the embryo grew, reaching 5.5 μl h⁻¹ at hatching. Throughout development, the capsule dimensions enlarged by absorption of water into the perivitelline space, increasing G _O2 by a combination of increasing surface area, and decreasing thickness of the capsule. These processes maintained P _O2in high enough to allow unrestricted V˙ _O2 until shortly before hatching. Diffusion limitation of respiration in hatching-stage embryos was demonstrated by (1) increased embryonic V˙ _O2 when P _O2out was experimentally raised, (2) greater V˙ _O2 of resting individuals immediately after hatching, and (3) reduced V˙ _O2 of hatchlings at experimental P _O2 levels higher than P _O2in before hatching. Thus, low P _O2in may be the stimulus to hatch. Potential problems of diffusive gas-exchange are mitigated by the relatively low incubation temperature (12 °C), which may be a factor limiting the distribution of the species to cool, southern waters. Received: 14 August 1999 / Accepted: 24 January 2000     

Mycosporine-like amino acids: possible UV protection in eggs of the sea hare Aplysia dactylomela

We investigated mycosporine amino acid (MAA) involvement as protective sunscreens in spawn of the sea hare Aplysia dactylomela to determine if adult diet and ultraviolet (UV) exposure affected the UV sensitivity of developing embryos. Adults were fed a red alga rich in MAAs (Acanthophora spicifera) or a green alga poor in MAAs (Ulva lactuca). Adults on each diet were exposed for 2 wk to ambient solar irradiance with two types of acrylic filters; one allowed exposure to wavelengths >275 nm (designated UV) and one to wavelengths only >410 nm (designated NOUV). Spawn from each adult group was likewise treated with UV or NOUV and monitored during development for differences in mortality and metabolic rate (measured as oxygen consumption: V˙ _O2). Also recorded were number of eggs or embryos per capsule, times to hatching, hatching success, size at hatching, and V˙ _O2 of adults. Spawn from adults eating red algae was almost twice as rich in MAAs as spawn from adults eating green algae, suggesting that MAA content is diet-related. Although overall quantities of MAAs in the spawn reflected MAA contents of the adult diet, specific MAAs were differentially sequestered in the spawn. Thus, porphyra-334, found in high concentration in Aplysia dactylomela's preferred red algal food, was present in only low concentration in the spawn. Conversely, mycosporine-glycine, in low concentration in red algal food, was the most abundant MAA in the spawn. UV treatment of adults had no effect on quantities of MAAs in the spawn. Adults exposed to UV had significantly higher V˙ _O2s and spawned twice as often. The UV-treated adults produced spawn with significantly higher V˙ _O2s and their embryos developed to hatching sooner. The only significant effect of UV exposure of the spawn was to reduce the percentage of veligers hatching from 71 to 50%. There was no significant effect on hatching time or size of the veligers at hatching, nor on number of eggs per capsule. Received: 13 May 1997 / Accepted: 27 June 1997     

Reproductive isolation and reproductive output in two sympatric mussel species (Mytilus edulis, M. trossulus) and their hybrids from Newfoundland

The mussels Mytilus edulis L. and M. trossulus Gould are found sympatrically in most areas of Newfoundland, with a low frequency of hybrids. To assess the potential for reproductive isolation, we sampled mussels from three sites in an eastern Newfoundland Bay from May–October 1996 to determine if there were differences in the reproductive cycles of the two species and their natural hybrids. In mussels with sheil lengths of 38–42 mm, males and females with mature gametes were dominant in June for M. edulis and hybrids, while M. trossulus showed a lower frequency of individuals with mature gametes. M. trossulus and hybrids spawned over a prolonged period (from late spring to early autumn) compared with most M. edulis individuals that spawned over a period of 2–3 weeks in July. This asynchrony in spawning activity between the two species may partially explain the low frequency of hybrids found in previous studies of these mussel populations. Female and male hybrids between M. edulis and M. trossulus showed normal gonad development, ripening and spawning, providing an opportunity for the introgression of genes between the two species. M. trossulus had a higher reproductive output than M. edulis of similar shell length, while hybrids showed intermediate values of reproductive output. M. trossulus females produced smaller eggs than either M. edulis or hybrids. Differences in reproductive traits may partially explain the maintenance of the mussel hybrid zone in Newfoundland. Published online: 13 August 2002     

Essential fatty acids influence metabolic rate and tolerance of hypoxia in Dover sole (Solea solea) larvae and juveniles

Dover sole (Solea solea, Linneaus 1758) were raised from first feeding on brine shrimp (Artemia sp.) with different contents and compositions of the essential fatty acids (EFA) arachidonic acid (ARA, 20:4n − 6); eicosapentaenoic acid (EPA, 20:5n − 3), and docosahexaenoic acid (DHA, 22:6n − 3), and their metabolic rate and tolerance to hypoxia measured prior to and following metamorphosis and settlement. Four dietary Artemia preparations were compared: (1) un-enriched; (2) enriched with a commercial EFA mixture (Easy DHA SELCO Emulsion); (3) enriched with a marine fish oil combination (VEVODAR and Incromega DHA) to provide a high ratio of ARA to DHA, and (4) enriched with these fish oils to provide a low ratio of ARA to DHA. Sole fed un-enriched Artemia were significantly less tolerant to hypoxia than the other dietary groups. Larvae from this group had significantly higher routine metabolic rate (RMR) in normoxia, and significantly higher O₂ partial pressure (PO₂) thresholds in progressive hypoxia for their regulation of RMR (P _crit) and for the onset of agitation, respiratory distress and loss of equilibrium. Metamorphosis was associated with an overall decline in RMR and increase in P _crit, but juveniles fed on un-enriched Artemia still exhibited higher P _crit and agitation thresholds than the other groups. Sole fed un-enriched Artemia had significantly lower contents of EFA in their tissues, both before and after settlement. Thus, enriching live feeds with EFA has significant effects on the respiratory physiology of sole early life stages and improves their in vivo tolerance to hypoxia. We found no evidence, however, for any effect of the ratio of ARA to DHA.     

Testing for relationships between individual crab behavior and metabolic rate across ecological contexts

Two classes of explanations for covariation between activity level and metabolic rate among conspecifics have been proposed. First, individual-level variation in activity exhibited during the measurement of metabolic rate should covary with routine metabolic rate because movement increases respiration (a methodological relationship). Second, energetic-based hypotheses posit relationships between individual activity measured under more natural conditions and baseline measures of metabolic rate, among other behavioral, physiological, and life-history traits (functional relationships). Here, we examined these potential relationships between individual behavior and metabolic rate in the mud crab (Panopeus herbstii). Specifically, we tested for (1) an effect of crab activity in metabolic chambers (activity_chamber) on routine metabolic rate (RMR), and (2) an effect of crab activity in mesocosms that mimicked field conditions (activity_mesocosm) on standard metabolic rate (SMR). To test for context dependence, we assessed both activity-metabolic rate relationships in the absence and presence of predation threat from toadfish (Opsanus tau) in the form of waterborne chemical cues. Individual variation in activity_mesocosm and RMR was repeatable over time. In support of a methodological relationship, individual differences in RMR were partially explained by crab activity_chamber. After accounting for this methodological relationship, individual SMR was inversely related to activity_mesocosm, supporting an allocation model that predicts behavior and baseline metabolic rate compete for finite energy reserves. We found no evidence of context dependence in either activity-metabolic rate relationship. Thus, our study emphasizes the importance of considering methodological artifacts in elucidating functional relationships between individual behavior and energetics.     

A molecular approach to the ecology of a mussel (Mytilus edulis – M. trossulus) hybrid zone on the east coast of Newfoundland, Canada

Mytilus trossulus Gould and M. edulis L. coexist in mixed populations in Atlantic Canadian waters. Although there is evidence that the two species hybridize in natural populations and that hybrids produce progeny through backcrosses, no study of the microgeographic distribution of the two forms and their hybrids has been made. Here we examine subtidal samples of mussels taken in July 1997 from two locations in eastern Newfoundland (Canada) and from wave-exposed and protected environments within each location. Shell lengths ranged from 15 to about 90 mm. Mussels were classified as pure forms or hybrids (F₁, F₂ and from backcrosses) based on four diagnostic markers, two allozyme loci (Mpi and Est-D) and two nuclear PCR-based DNA markers (ITS and Glu-5). In addition, a PCR-based mtDNA marker (COIII) was used to characterize the distribution of mtDNA mitotypes among pure and hybrid individuals. There were differences in the proportions of pure M. edulis and M. trossulus between sites and between environments at one location. M. trossulus was the predominant species at one of the two exposed sites. In all four samples, M. trossulus was also the predominant form among small individuals. The frequency of hybrids was 26% overall and did not differ among samples. Hybrids consisted mostly of backcrosses that were M. trossulus-biased among small mussels and M. edulis-biased among large ones. We conclude that both intrinsic genetic factors and extrinsic environmental factors influence the relative frequency of M. edulis, M. trossulus and their hybrids. Received: 29 June 1998 / Accepted: 4 November 1998     

Heavy metal contamination and physiological variability in the Brazilian mangrove crabs Ucides cordatus and Callinectes danae (Crustacea: Decapoda)

 Physiological studies were made on the crabs Ucides cordatus (L.) and Callinectes danae sampled from populations living in “polluted” mangroves on the southeast littoral of Brazil. Analysis of Cu, Cd, Zn, and Fe of sediments and crab tissues showed interspecific differences in tissue concentrations, and significantly higher levels of Cu, Cd, and Zn in “polluted” populations compared to “unpolluted” crabs living in uncontaminated mangrove in the same geographical area. Individuals of both species from the polluted site showed significantly greater capacities for regulating blood osmotic concentrations at low salinity (9‰). However, U. cordatus showed a reduced hypo-regulatory ability in 34‰S. Differences in ionoregulation were also seen. “Polluted”C. danae showed significantly higher Na/ K-ATPase levels in posterior gills compared to “unpolluted” crabs. Oxygen consumption rates (M˙ _O2) were elevated in U. cordatus, but depressed in C. danae from the “polluted” population. Individuals of both species from this site showed significantly lower O:N ratios, mainly because of an increased net efflux of ammonia. Adenylate energy charge (AEC) values of muscle and hepatopancreas in “unpolluted” and “polluted” populations of both species were not significantly different. These physiological differences are discussed in relation to the known acute physiological and metabolic effects of heavy metals in crustaceans, and interpretated in the light of possible adaptive changes following long-term exposure to contamination. Received: 6 August 1999 / Accepted: 22 June 2000     

Aerobic metabolic rates of swimming juvenile mako sharks, <Emphasis Type="Italic">Isurus oxyrinchus</Emphasis>

The shortfin mako shark, Isurus oxyrinchus, is a highly streamlined epipelagic predator that has several anatomical and physiological specializations hypothesized to increase aerobic swimming performance. A large swim-tunnel respirometer was used to measure oxygen consumption (MO₂) in juvenile mako sharks (swimming under controlled temperature and flow conditions) to test the hypothesis that the mako shark has an elevated maintenance metabolism when compared to other sharks of similar size swimming at the same water temperature. Specimen collections were conducted off the coast of southern California, USA (32.94°N and 117.37°W) in 2001-2002 at sea-surface temperatures of 16.0–21.0°C. Swimming MO₂ and tail beat frequency (TBF) were measured for nine mako sharks [77–107 cm in total length (TL) and 4.4 to 9.5 kg body mass] at speeds from 28 to 54 cm s⁻¹ (0.27–0.65 TL s⁻¹) and water temperatures of 16.5–19.5°C. Standard metabolic rate (SMR) was estimated from the extrapolation to 0-velocity of the linear regression through the LogMO₂ and swimming speed data. The estimated LogSMR (±SE) for the pooled data was 2.0937 ± 0.058 or 124 mg O₂ kg⁻¹ h⁻¹. The routine metabolic rate (RMR) calculated from seventeen MO₂ measurements from all specimens, at all test speeds was (mean ± SE) 344 ± 22 mg O₂ kg⁻¹h⁻¹ at 0.44 ± 0.03 TL s⁻¹. The maximum metabolic rate (MMR) measured for any one shark in this study was 541 mg O₂ kg⁻¹h⁻¹ at 54 cm s⁻¹ (0.65 TL s⁻¹). The mean (±SE) TBF for 39 observations of steady swimming at all test speeds was 1.00 ± 0.01 Hz, which agrees with field observations of 1.03 ± 0.03 Hz in four undisturbed free-swimming mako sharks observed during the same time period. These findings suggest that the estimate of SMR for juvenile makos is comparable to that recorded for other similar-sized, ram-ventilating shark species (when corrected for differences in experimental temperature). However, the mako RMR and MMR are apparently among the highest measured for any shark species.     

Longevity,calling effort,and metabolic rate in two populations of cricket

Intraspecific variation in a resting metabolic rate (RMR) is likely to be an important determinant of energetic-resource use and may influence the resources subsequently available for allocation to traits not directly associated with somatic maintenance. The influence of RMR on resource availability could be especially important for condition-dependent sexual traits, such as cricket calls, that are themselves energetically costly to produce. RMR may also be associated with longevity, either negatively because individuals with a high RMR burn resources faster and die young, or positively as individuals with high RMR are more able to accrue resources to fuel survival. Additionally, the associations between RMR and other characters may vary across populations if differential selection or drift shapes these traits. Here we tested for differences in RMR, body mass, calling effort, and longevity in two populations of cricket Gryllodes sigillatus and then evaluated the potential influence of RMR on calling and longevity. We find that RMR, calling effort, and longevity varied across populations, but mass did not. Controlling for population and mass, RMR was not significantly associated with calling effort, but was negatively associated with longevity. These findings suggest that male crickets that live fast die young.     

A possible hybrid zone in the Mytilus edulis complex in Japan revealed by PCR markers

The adhesive protein allele of mussels collected at 13 points in Japan from Hokkaido to Kyushu was analyzed by the polymerase chain reaction using a set of primers which amplifies a part of the nonrepetitive region of the adhesive protein gene. While most mussels exhibited a 126 bp fragment, characteristic of the pure Mytilus galloprovincialis, 55 of 64 mussels sampled at Hiura and 1 of 14 mussels at Hakodate Port exhibited 168 and 126 bp fragments. Sequence analysis of the two fragments indicated that the 168 and 126 bp fragments are almost identical to previously reported sequences in M. trossulus and M. galloprovincialis, respectively. Since the frequency of heterozygous individuals in Hiura is very high, it is unlikely that they are simple hybrids. However, it is evident that mixing of genes occurred between the two species off Hokkaido. Received: 6 September 1996 / Accepted: 9 October 1996     

Diurnal cycle and kinetics of ammonium assimilation in the green alga <Emphasis Type="Italic">Ulva pertusa</Emphasis>

The kinetics of ammonium assimilation was investigated in Ulva pertusa (Chlorophyceae, Ulvales) from northeastern New Zealand. Ammonium assimilation exhibited Michaelis–Menten kinetics with a maximum rate of assimilation (V _max) of 54 ± 5 μmol g⁻¹ dry weight h⁻¹ and half-saturation constant (K _m) of 23 ± 8 μM. In contrast, values for ammonium uptake were considerably higher with a V _max of 316 ± 59 μmol g⁻¹ dry weight h⁻¹ and K _m of 135 ± 46 μM. At environmentally relevant ammonium concentrations (5 μM), assimilation accounted for most (70%) of the ammonium taken up. Darkness decreased the maximum rate of ammonium assimilation by 83%. We investigated the hypothesis that rates of biosynthetic processes are greater in the early part of the day in Ulva. Consistent with this hypothesis, the maximum rate of ammonium assimilation in U. pertusa peaked in the morning and coincided with low levels of the photosynthetic product sucrose, which peaked in the afternoon. There was a diurnal cycle in the rate of ammonium uptake and assimilation in light and dark, but the amplitude was much greater for assimilation than uptake. Moreover, our data suggest that net ammonium assimilation only occurs during the day in U. pertusa. We suggest that two major roles for diurnal cycles are minimisation of interspecific competition for resources and metabolic costs.     

Tolerance to low oxygen and sulfide in Amphiura filiformis and Ophiura albida (Echinodermata: Ophiuroidea)

The brittle stars Amphiura filiformis (Müller) and Ophiura albida (Forbes) were exposed to different oxygen saturations (100, 10, 5, 3, and <1% oxygen saturation) and to physiological anoxia (<1% oxygen saturation) at different total sulfide concentrations (0, 2, 20, 200 μM). The mortality was followed during experiments and the median survival time (LT₅₀) was determined. The infaunal A. filiformis had a significantly higher tolerance to both hypoxia and sulfide than did the epibenthic O. albida. After exposure to 10% oxygen saturation for a month, only 2.0% A. filiformis and 0% O. albida were dead. In oxygen saturations <1% A. filiformis and O. albida had a LT₅₀ of 7.5 and 2.5 d, respectively. The presence of even very small concentrations of sulfide decreased the survival significantly. Sulfide is shown to be the key factor for the survival of the two species. Received: 11 October 1996 / Accepted: 12 November 1996     

Effect of MS-222 on response to light and rate of metabolism of the little skate Raja erinacea

The effect of tricaine methanesulfonate (MS-222) on the standard (SMR) and routine (RMR) metabolic rates of Raja erinacea was estimated from oxygen-consumption measurements. Data were gathered from a computerized, flow-through respirometry system. Individual trials were run for 96 h at 10 °C on anesthetized and untreated fish. The resulting rates, SMR = 20.1 ± 1.99 SE and RMR = 48.3± 2.5 SE mg O₂ kg⁻¹ h⁻¹, for an unanesthetized skate of 0.5 kg standardized weight, are the lowest reported for any elasmobranch. Periodogram analysis revealed a significant light-response component to the oxygen consumption of these fish, indicating a pattern of nocturnal and crepuscular activity. This activity pattern was disrupted in skates subjected to anesthesia. The use of low-dosage MS-222 in conjunction with the respirometry trials provides positive preliminary evidence that this technique may be useful in giving quick, accurate estimates of SMR in the more intractable elasmobranch fishes. Received: 15 January 1997 / Accepted: 6 February 1997     

Energetics of underwater swimming in the great cormorant (Phalacrocorax carbo sinensis)

Resting metabolic rate (RMR), energy requirements and body core temperature were measured during underwater swimming in great cormorants (Phalacrocorax carbo sinensis) at the zoological garden in Neumünster, Germany, using gas respirometry and stomach temperature loggers. We used a 13 m long still water canal equipped with a respiration chamber at each end. Birds swam voluntarily in the canal at a mean speed of 1.51 ms^-1. Power input during underwater swimming averaged 31.4 W kg^-1. Minimal costs of transport of 19.1 J kg^-1 m^-1 were observed at a speed of 1.92 m s^-1. Body core temperature was stable in all birds within the first 60 min spent in the canal. After that, body temperature dropped at a rate of 0.14°C min^-1 until the birds voluntarily left the water. Our data indicate that great cormorants spend 2.7 times more energy than Adélie penguins (Pygoscelis adeliae) during underwater swimming. This can be essentially attributed to their poor insulation, their mode of locomotion underwater and differences in streamlining. RMR on land was related to body mass via VO₂=0.691 M^0.755 (where VO₂ is O₂-consumption in litre h^-1 and M is body mass in kg). In order to quantify the effects of external devices on energy consumption during underwater swimming, we tested a dummy data logger attached to the back of the cormorants as well as a ring on the leg. The ring had no apparent influence on the swimming energetics of the cormorants. In birds equipped with dummy loggers, swimming speed was not significantly influenced, but both power input and costs of transport increased by a mean of 19% for swimming speeds between 1.4 and 1.8 m s^-1.     

Carbon-dioxide production and metabolic parameters in the ophiurid Ophiothrix fragilis

As part of the evaluation of fluxes between the water column and a rich benthic community of the Dover Strait (Eastern English Channel), laboratory measurements of oxygen consumption were carried out on a common ophiurid, Ophiothrix fragilis (Abildgaard), from February 1993 to February 1995. The mean O₂-consumption rate was evaluated at 0.31 mg O₂ g⁻¹ h⁻¹ (ash-free dry weight). Simultaneous measurements of O₂ consumption and CO₂ production using the pH-alkalinity method revealed an average respiratory quotient of 0.69 proved suitable for converting oxygen demand to carbon flux. A seasonal trend in respiration data was demonstrated by sinusoidal curves fitted to O₂-uptake and CO₂-release data as a function of time. The influence on respiration rate of two seasonal parameters (temperature and food availability) is discussed; linear regression indicated a highly significant relationship between O₂ consumption (or CO₂ production) and temperature; both O₂-consumption and CO₂-production rates decreased with starvation. The average O:N ratio was estimated at 8.46, close to the theoretical value when proteins constitute the catabolic substrate. The annual carbon respired by the O. fragilis community examined and the estimated annual primary production by phytoplankton indicate that the respiration of the O. fragilis community could supply 35% of phytoplankton carbon requirements. Received: 1 August 1996 / Accepted: 4 September 1996     

Haemolymph constituent levels and ammonia efflux rates of Nephrops norvegicus during emersion

Nephropsnorvegicus (L.) were subjected to 8 h of emersion, either between layers of seawater-soaked hessian with periodical (20 min) flushes of seawater (high humidity, HH) or to unprotected emersion (low humidity, LH). Blood ammonia levels rose during emersion in both groups but reached higher levels under LH conditions. Ammonia efflux rates after re-immersion were higher than those of control prawns, and amounts of ammonia excreted at such times were considerably higher than those calculated to have accumulated in the blood during emersion. Possible explanations for such differences are discussed. C _aO2 and C _vO2 decreased rapidly to ca. 10% normoxia values within 2 h of HH and LH emersion and remained low throughout the remaining emersion time. Emersion-induced tissue hypoxia increased blood concentrations of glucose and lactate. Lactate accumulation was higher during LH emersion, compared with HH emersion. Blood pH dropped ca. 0.40 units but increased again after 2 h of re-immersion. Acidosis was probably related more to respiratory difficulties (CO₂ accumulation) than to lactate accumulation, as blood lactate values remained high after 2 h of re-immersion. The ability of N. norvegicus to cope with emersion appears to be little influenced by high humidity conditions. Received: 26 June 1996 / Accepted: 5 August 1996     

Diel cycles of activity,metabolism, and ammonium concentration in tropical holothurians

Movement rate, oxygen consumption, and respiratory tree ammonium concentration were measured in situ in the holothurians Pearsonothuria graeffei and Holothuria edulis in the Agan-an Marine Reserve, Sibulan, Philippines (9°20′30″N, 123°18′31″E). Measurements were made both day and night for both species during April–July 2005. P. graeffei had significantly higher movement rate during the day than at night (1.14 and 0.27 m h⁻¹, respectively; three-way ANOVA, P < 0.05) while H. edulis had higher movement rate at night compared to the day (0.83 and 0.07 m h⁻¹, respectively), spending the daylight hours sheltering under coral. More than 80% of H. edulis had movement rate of zero during the day. Oxygen consumption of P. graeffei was significantly higher during the day than at night (1.61 and 0.83 μmol O₂ g⁻¹ h⁻¹, respectively; two-way ANCOVA, P < 0.05), but the reduction at night was not as pronounced as the reduction in movement. H. edulis had a 75% reduction in oxygen consumption during the day compared to night (0.51 and 1.96 μmol O₂ g⁻¹ h⁻¹, respectively), matching this species’ reduced movement rates during the day. Ammonium concentration in water withdrawn from the respiratory trees of P. graeffei during the day (12.0 μM) was three times higher than in respiratory tree water sampled at night (4.3 μM) and 15 times higher than ambient seawater (0.8 μM; three-way ANOVA, P < 0.05). Ammonium concentration in the respiratory tree water of H. edulis was six times higher at night (14.6 μM) than during the day (2.2 μM) and 16 times higher than that of ambient seawater (0.9 μM). Even though H. edulis and P. graeffei are found within the same coral reef environment, they may affect different substrates and reef organisms due to their different habitats and distinct but opposite diel cycles.     

Genetic structure and allozyme variation of sea bass (Dicentrarchus labrax and D. punctatus) in the Mediterranean Sea

This paper reports data on 28 allozyme loci in wild and artificially reared sea bass (Dicentrarchus labrax) samples, originating from either coastal lagoon or marine sites in the Mediterranean Sea. F _ST analysis (θ estimator) indicated strong genetic structuring among populations; around 34% of the overall genetic variation is due to interpopulation variation. Pairwise θ estimates showed that, on average, the degree of genetic structuring was much higher between marine populations than between samples from lagoons. Six polymorphic loci showed differences in allele frequencies between marine and lagoon samples. Multivariate analyses of individual allozymic profiles and of allele frequencies suggested that different arrays of genotypes prevail in lagoons compared to marine samples, particularly at those loci that, on the basis of previous acclimation experiments, had been implicated in adaptation to freshwater. On the other hand, variation at “neutral” allozyme loci reflects to a greater extent the geographic location of populations. Allozyme differentiation was also studied in a D. labrax population from the Portuguese coast. Average genetic distance between this population and the Mediterranean populations was quite high (Nei's D = 0.236) and calls into question the taxonomic status of the Portuguese population. Finally, genetic relationships between D. labrax and D. punctatus were evaluated. Average Nei's D was 0.648, revealing high genetic differentiation between the two species, even for two sympatric populations of these species in Egypt; thus gene flow was not indicated between species. Received: 24 October 1996 / Accepted: 27 November 1996     

Low-temperature adaptation of deep-sea bacteria isolated from the Japan Trench

Montastraea annularis, M. faveolata, and M.␣franksi are three recently separated species that together dominate reefs of the tropical western Atlantic. Despite morphological, life-history, ecological and genetic differences, the legitimacy of their status as separate species has been questioned. This controversy stems from both the scarcity of unambiguous, diagnostic differences among them, and from the possibility of extensive hybridization associated with their approximately synchronous reproduction in sympatry. Here we report on fertilization trials and the timing of spawning, both of which suggest that the potential for hybridization may be limited. Crosses between M. faveolata (the most genetically distinctive taxon) and the other two species were largely unsuccessful, as were selfed matings for all three species. M. annularis and M. franksi showed no evidence of fertilization barriers, but the timing of spawning typically differed between them by 1 to 2 h. We also found that spawning times in the field of M. annularis and M. faveolata were non-overlapping in 1995, and that the timing of spawning in M. annularis could be experimentally shifted forward by simulating earlier than natural sunsets. These findings from Panamá and Honduras, particularly given their consistency with comparable observations and experiments elsewhere, provide evidence of reproductive isolation and support the separate species status of these three taxa. Some hybridization may occur under natural conditions at these sites, particularly between M. annularis and M.␣franksi. Rampant genetic interchange among these taxa in Central America seems unlikely, however, in the light of the concordant associations between morphology, reproductive biology and other characters which are observed. Received: 22 July 1996 / Accepted: 2 October 1996     

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