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     

Protection of Genetic Diversity and Maintenance of Connectivity among Reef Corals within Marine Protected Areas

Abstract: High‐latitude coral reefs (HLRs) are potentially vulnerable marine ecosystems facing well‐documented threats to tropical reefs and exposure to suboptimal temperatures and insolation. In addition, because of their geographic isolation, HLRs may have poor or erratic larval connections to tropical reefs and a reduced genetic diversity and capacity to respond to environmental change. On Australia's east coast, a system of marine protected areas (MPAs) has been established with the aim of conserving HLRs in part by providing sources of colonizing larvae. To examine the effectiveness of existing MPAs as networks for dispersal, we compared genetic diversity within and among the HLRs in MPAs and between these HLRs and tropical reefs on the southern Great Barrier Reef (GBR). The 2 coral species best represented on Australian HLRs (the brooding Pocillopora damicornis and the broadcast‐spawning Goniastrea australensis) exhibited sharply contrasting patterns of diversity and connectedness. For P. damicornis, the 8‐locus genetic and genotypic diversity declined dramatically with increasing latitude (N_a= 3.6–1.2, H_e= 0.3–0.03, N_g:N = 0.87–0.06), although population structure was consistent with recruitment derived largely from sexual reproduction (G_o:G_e= 1.28–0.55). Genetic differentiation was high among the HLRs (F_ST[SD]= 0.32 [0.08], p < 0.05) and between the GBR and the HLRs (F_ST= 0.24 [0.06], p < 0.05), which indicates these temperate populations are effectively closed. In contrast for G. australensis, 9‐locus genetic diversity was more consistent across reefs (N_a= 4.2–3.9, H_e= 0.3–0.26, N_g:N = 1–0.61), and there was no differentiation among regions (F_ST= 0.00 [0.004], p > 0.05), which implies the HLRs and the southern GBR are strongly interconnected. Our results demonstrate that although the current MPAs appear to capture most of the genetic diversity present within the HLR systems for these 2 species, their sharply contrasting patterns of connectivity indicate some taxa, such as P. damicornis, will be more vulnerable than others, and this disparity will provide challenges for future management.     

Mitochondrial DNA analysis of the genetic relationships among populations of scad mackerel (Decapterus macarellus, D. macrosoma, and D. russelli) in South-East Asia

The genetic relationships among South-East Asian populations of the scad mackerels Decapterus macarellus, D. macrosoma and D. russelli (Pisces: Carangidae) were investigated. In 1995 and 1996, 216 fish were sampled in seven localities spanning the seas of Indonesia and were examined for restriction-site polymorphisms using ten restriction enzymes for the mitochondrial (mt) DNA control region, amplified by the polymerase chain-reaction. The inferred phylogeny of haplotypes led to the recognition of three distinct mitochondrial lineages or phylads consistent with the distinctions of current taxonomy. All 15 mtDNA haplotypes found in D. macarellus and all 9 haplotypes found in D. macrosoma were arranged as star-like clusters, suggesting recent evolutionary history. In contrast, the phylad formed by 6 haplotypes in D. russelli from the Sulawesi Sea exhibited diffuse topology, suggesting that ancestral lineages of this species have been retained to the present. Average nucleotide-divergence estimates between haplotypes of different phylads were between 0.042 and 0.135, suggesting ancient separation, in consistency with published allozyme data. High levels of haplotype diversity, but no geographical heterogeneity, was detected within D. macarellus from the Molucca Sea and the Banda Sea. Populations of D. macrosoma exhibited both significant differences between adjacent regions (Sunda Strait and Java Sea), and broadscale genetic homogeneity from the South China Sea to the Sulawesi Sea via the Java Sea and Makassar Strait. The geographic isolation of the D. macrosoma population sampled in the Sunda Strait suggests that this region constitute a sharp transition zone between the Indian Ocean and the Sunda Shelf. Near-monomorphism of haplotypes and low nucleotide diversity (d _X) were observed in the samples of D. macrosoma from the continental shelf (haplotype-diversity estimates, h, = 0.00 to 0.25 ± 0.08 and d _X = 0.000 to 0.002). This was in contrast to the comparatively high haplotype and nucleotide diversities observed in other pelagic fish species including D. macarellus (h = 0.82 ± 0.05, d _X = 0.012 to 0.015) and D. russelli (h = 0.63 ± 0.12, d _X = 0.016), and in the oceanic D. macrosoma population sampled in the Sunda Strait (h = 0.67 ± 0.31, d _X = 0.005). We hypothesise that this may be the consequence of recent and perhaps repeated bottleneck events that have affected the D. macrosoma population sampled on the continental shelf. Received: 29 September 1997 / Accepted: 3 September 1999     

Sociality,Bateman’s gradients,and the polygynandrous genetic mating system of round-tailed ground squirrels (<Emphasis Type="Italic">Xerospermophilus tereticaudus</Emphasis>)

Historically, most mammals have been classified as polygynous; although recent molecular evidence suggests that many mammals may be polygynandrous, particularly the ground-dwelling sciurids. We genotyped 351 round-tailed ground squirrels (Xerospermophilus tereticaudus) using seven microsatellite loci to determine paternity in 31 litters from 2004 to 2007. Polygyny was evident in all years except in 2007, when the population size was reduced. Multiple paternity occurred in the majority of litters (55%) with 2.5 ± 0.26 sires/litter (n = 31). Forty-nine percent of resident males (n = 114) sired offspring, and of males that sired offspring (n = 56) 27% sired young in multiple litters in a single breeding season. Litter size was positively correlated with the number of sires. Through an indirect analysis of paternity, we found 21 litters (68%) with an average relatedness of 0.5 or less. Males had a greater opportunity for sexual selection (I _s = 1.60) than females (I _s = 0.40); Bateman’s gradient was also greater in males (1.07 ± 0.04, n = 56) than females (0.82 ± 0.08, n = 31). The mating system in round-tailed ground squirrels defined through genetic analyses and Bateman’s gradients is polygynandrous compared to the previously suggested polygynous mating system as established by behavioral observations and fits within the predictions of the ground squirrel sociality models. Upon evaluating the predictions of the sociality models among sciurid species, we found a negative relationship between the level of sociality with litter size and the average percentage of multiple paternity within a litter. Thus, recent genetic information and reclassification of mating systems support the predictions of the ground-dwelling squirrel sociality models.     

Genetic study of the extent and consequences of sexual and asexual reproduction in the deep-sea epizoic anemones Amphianthus inornata and Kadosactis commensalis (Cnidaria: Anthozoa)

The anemone Amphianthus inornata is found at bathyal depths living on colonies of the gorgonian Acanella arbuscula. Previous studies of the morphology and reproductive stage of this anemone, during different times of the year, have indicated that it reproduces sexually on a seasonal basis. A small proportion of the study population were also reported to be undergoing asexual reproduction by fission. The anemone Kadosactis commensalis is also bathyal, but lives mainly on the holothurian Paroriza prouhoi. Previous morphological studies have indicated that K. commensalis is a protandrous hermaphrodite that exhibits non-seasonal sexual reproduction only. In the present study, allozyme electrophoresis was used to examine the prevalence and genetic consequences of asexual reproduction in a population of Amphianthus inornata from 2 200 m in the Rockall Trough, North Atlantic Ocean. Genetic evidence, from five randomly selected polymorphic enzyme loci, for asexual reproduction in this species was weak. Exact tests indicated that genotype frequencies did not differ significantly from those expected under Hardy–Weinberg equilibrium. F _IS (correlation of homologous alleles with reference to local population, assuming random mating) values also did not differ significantly from zero, and observed heterozygosity (H _o =0.446) and genotypic diversity (G _o =17.0387) were very similar to Hardy–Weinberg expected frequencies (H _e =0.446; G ^* _e =17.0010). Evidence suggests that the contribution of asexual reproduction to recruitment in the study population of A. inornata is low. For a single population of K. commensalis from 4 850 m on the Porcupine Abyssal Plain, North Atlantic Ocean, the hypothesis that inbreeding due to reduced occurrence of outcrossing between anemones on a single holothurian host was examined by electrophoresis of ten randomly selected enzyme loci. Single-locus genotypic frequencies were significantly different from expected frequencies for one locus P≤ 0.05, hexokinase-1 (Hex-1)]. F _IS values were significantly different from zero for two enzyme loci (Hex-1 and Hex-2, P≤ 0.01 and P≤ 0.05, respectively), and the overall observed heterozygosity was lower than the expected heterozygosity (H _o =0.125, H _e =0.140). The hypothesis of inbreeding could not be rejected by the present study, although sample size was small (N=55), leading to possible bias in tests of significance. Genetic variation in A. inornata was higher than that recorded for most eukaryotes, although interlocus sampling error for only five loci is high. High genetic variability has been found in other sea anemones, and has been related to high longevity and mixed reproductive modes. Genetic variation in K. commensalis was in the higher range of that found in other eukaryotes, and is not unusual for anemones. Received: 5 August 1996 / Accepted: 11 December 1996     

Population genetics of the nurse shark (Ginglymostoma cirratum) in the western Atlantic

The nurse shark, Ginglymostoma cirratum, inhabits shallow, tropical, and subtropical waters in the Atlantic and the eastern Pacific. Unlike many other species of sharks, nurse sharks are remarkably sedentary. We assayed the mitochondrial control region and eight microsatellite loci from individuals collected primarily in the western Atlantic to estimate the degree of population subdivision. Two individuals from the eastern Atlantic and one from the Pacific coast of Panama also were genotyped. Overall, the mtDNA haplotype (h = 48 ± 5%) and nucleotide (π = 0.08 ± 0.06%) diversities were low. The microsatellite data mirror the mitochondrial results with the average number of alleles ([`(N)]<sub>A</sub> \bar{N}_{A}  = 9) and observed heterozygosity ([`(H)]_O \bar{H}_{O}  = 0.58) both low. The low levels of diversity seen in both the mtDNA and the microsatellite may be due to historical sea level fluctuations and concomitant loss of shallow water habitat. Eight of the 10 pair-wise western Atlantic F _ST estimates for mtDNA indicated significant genetic subdivision. Pair-wise F _ST values for the microsatellite loci indicated a similar pattern as the mtDNA. The western Atlantic population of nurse sharks is genetically subdivided with the strongest separation seen between the offshore islands and mainland Brazil, likely due to deep water acting as a barrier to dispersal. The eastern and western Atlantic populations were closely related. The eastern Pacific individual is quite different from Atlantic individuals and may be a cryptic, sister species.     

Icy heritage: ecological evolution of the postglacial Baltic Sea reflected in the allozymes of a living fossil,the priapulid Halicryptus spinulosus

Genetic variation of 16 allozyme loci in 397 Halicryptus spinulosus (Priapulida) revealed overall polymorphism of P=0.438 and Hardy-Weinberg expectations for heterozygosity of H _e=0.060 for Baltic Sea stocks, H _e=0.143 for the White Sea and H _e=0.121 for Iceland. Maximal unbiased standard distances of D=0.0693 separated Baltic and White Sea populations. Nordic and Baltic populations could be distinguished by allozymes, but Baltic subsamples proved cohesive. Gene flow amounted to effective exchange values per generation of N _m=2.94 over 650 km of continuous habitat, N _m=10.65 over 175 km, and N _m=13.85 over 20 km. Gene flow started to decrease with geographic distance beyond a dispersal threshold of 20 km, but hierarchical G _ST-statistics indicated light isolation by distance beyond a minimum of 8 km. Gene flow is high for a benthic worm assumed to lack dispersal by pelagic larva, a paradox which cannot be resolved now. Baltic populations are characterized by lower heterozygosity than Nordic stocks. In the Baltic Basin, temporally continuous brackish-water conditions have only existed for the past 7000 years. The two possible colonization routes of H. spinulosus to the geologically young Baltic Sea imply genetic drift, whether by founder effect (sweepstake colonization from Iceland) or by refugial bottlenecking during the Ancylus phase of the Baltic Basin after a direct connection to the White Sea had been sequestered. Continued genetic drift is emphasized by lower heterozygosity in the ecologically unstable Belt Sea compared to the central Baltic. Allozymes falsify the reduced-mutability hypothesis to explain bradytelic evolution of Priapulida. Regional genetic homogeneity, ample polymorphism, and preference for anoxic black mud qualify H. spinulosus populations as indicators of microevolutionary responses to water circulation regimes or pollution in the Baltic Sea.     

Cryptic species and population structuring of the Atlantic and Pacific seabob shrimp species, Xiphopenaeus kroyeri and Xiphopenaeus riveti

Seabob shrimps of the genus Xiphopenaeus are important fishery resources along the Atlantic and Pacific coasts of Central and South America. The genus was considered to comprise two species: the Atlantic Xiphopenaeus kroyeri (Heller, Sitzungsber Math Naturwiss cl kaiserliche Akad Wiss Wien 45:389–426, 1862), and the Pacific Xiphopenaeus riveti (Bouvier, Bull Mus Hist Nat Paris 13:113–116, 1907). In a recent review, Xiphopenaeus was regarded as a monotypic genus, on the basis that no clear morphological differences could be found between Pacific and Atlantic specimens (Pérez Farfante and Kensley, Mem Mus Nat Hist Nat Paris 175:1–79, 1997). In the present work, nuclear (allozymes), and mitochondrial (Cytochrome Oxidase I) genes were used to demonstrate the validity of X. riveti and reveal the presence of two cryptic species of Xiphopenaeus within X. kroyeri in the Atlantic Ocean. The high levels of molecular divergence among these species contrast with their high morphological resemblance. Interspecific sequence divergences (Kimura 2-parameter distance) varied from 0.106 to 0.151, whereas intraspecific distances ranged from 0 to 0.008 in Xiphopenaeus sp. 1, from 0 to 0.003 in Xiphopenaeus sp. 2, and from 0.002 to 0.005 in X. riveti. In addition, five diagnostic allozyme loci were found between sympatric samples of Xiphopenaeus sp. 1 and 2 along the Brazilian coast. The results suggest that Xiphopenaeus sp. 2 from the Atlantic is more closely related to the Pacific X. riveti than to the Atlantic Xiphopenaeus sp. 1. Furthermore, a high level of genetic structuring (Xiphopenaeus sp. 1: F _ST =0.026; P<0.05; Xiphopenaeus sp. 2: F _ST =0.055; P<0.01) was found in the Brazilian Xiphopenaeus populations, indicating the presence of different genetic stocks in both Atlantic species. These findings have important commercial implications as they show that the fisheries of the two Atlantic species must be managed separately, and that each one is comprised of different populations.Communicated by O. Kinne, Oldendorf/Luhe     

Genetic evidence for contrasting patterns of dispersal in solitary and colonial ascidians

In situ and in vitro observations indicate that brooding colonial ascidians commonly display limited larval dispersal, whilst the larvae of most solitary species are assumed to be widely dispersed. We used allozyme data to determine the population genetic consequences of reproduction and dispersal in a broadcast-spawning solitary ascidian and two brooding colonial species along the central and southern coast of New South Wales, Australia. We surveyed genetic variation at 2 to 9 variable loci for samples collected from 6 to 8 local populations of each of the stalked solitary species Pyura gibbosa gibbosa Heller, 1878; the social Stolonica australis Michaelsen, 1927 and the compound Botrylloides magnicoecum Hartmeyer, 1912. Samples from each local population displayed levels and patterns of genotypic diversity that were consistent with expectations for sexually-derived recruitment of both solitary zooids and separate colonies. However, we found clear differences in the structure of the populations of solitary and colonial species. Genotype frequencies within all nine samples of P. gibbosa gibbosa conformed to expectations for random mating (i.e. Hardy–Weinberg equilibria). Moreover, allele frequencies showed little variation among samples [mean standardised genetic variance (F _{S T} ) =0.002], which implies that local populations are strongly connected by larval dispersal. We estimate (via Wright's “island model”) that gene flow (N _e m) within this set of local populations is 125 effective migrants per generation, which is very similar to estimates obtained for other broadcast-spawning taxa in this region. In contrast, genotype frequencies within samples of both colonial species were characterised by large and statistically significant deficits of heterozygotes, consistent with expectations for highly limited dispersal of larvae or sperm. Moreover, local populations were highly differentiated (F _{S T} =0.201 and 0.202 for S. australis and B. magnicoecum, respectively) and N _e m was estimated to be ∼1.0 in each case. These values of F _{S T} and subsequent estimates of N _e m lie within the range of values reported for other New South Wales taxa with direct larval development, and imply that local populations are effectively closed to immigration. Received: 13 February 1997 / Accepted 18 July 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     

Population structure,mortality and growth of <Emphasis Type="Italic">Pinna nobilis</Emphasis> Linnaeus, 1758 (Mollusca,Bivalvia) at different depths in Moraira bay (Alicante,Western Mediterranean)

An investigation to characterize the causes of Pinna nobilis population structure in Moraira bay (Western Mediterranean) was developed. Individuals of two areas of the same Posidonia meadow, located at different depths (A1, −13 and A2, −6 m), were inventoried, tagged, their positions accurately recorded and monitored from July 1997 to July 2002. On each area, different aspects of population demography were studied (i.e. spatial distribution, size structure, displacement evidences, mortality, growth and shell orientation). A comparison between both groups of individuals was carried out, finding important differences between them. In A1, the individuals were more aggregated and mean and maximum size were higher (A1, 10.3 and A2, 6 individuals/100 m²; A1, x = 47.2 ± 9.9; A2, x = 29.8 ± 7.4 cm, P < 0.001, respectively). In A2, growth rate and mortality were higher, the latter concentrated on the largest individuals, in contrast to A1, where the smallest individuals had the higher mortality rate [A1, L = 56.03(1 − e^−0.17t ); A2, L = 37.59(1 − e^−0.40t ), P < 0.001; mean annual mortality A1: 32 dead individuals out of 135, 23.7% and A2: 16 dead individuals out of 36, 44.4%, and total mortality coefficients (z), z _A1(−30) = 0.28, z _A1(31–45) = 0.05, z _A1(46−) = 0.08; z _A2(−30) = 0.15, z _A2(31–45) = 0.25]. A common shell orientation N–S, coincident with the maximum shore exposure, was observed in A2. Spatial distribution in both areas showed not enough evidence to discard a random distribution of the individuals, despite the greater aggregation on the deeper area (A1) (A1, χ ² = 0.41, df = 3, P > 0.5, A2, χ ² = 0.98, df = 2 and 0.3 < P < 0.5). The obtained results have demonstrated that the depth-related size segregation usually shown by P. nobilis is mainly caused by differences in mortality and growth among individuals located at different depths, rather than by the active displacement of individuals previously reported in the literature. Furthermore, dwarf individuals are observed in shallower levels and as a consequence, the relationship between size and age are not comparable even among groups of individuals inhabiting the same meadow at different depths. The final causes of the differences on mortality and growth are also discussed.     

Microsatellite analysis of albacore tuna (<Emphasis Type="Italic">Thunnus alalunga</Emphasis>): population genetic structure in the North-East Atlantic Ocean and Mediterranean Sea

Stock heterogeneity was investigated in albacore tuna (Thunnus alalunga, Bonnaterre 1788), a commercially important species in the North Atlantic Ocean and Mediterranean Sea. Twelve polymorphic microsatellite loci were examined in 581 albacore tuna from nine locations, four in the north-east Atlantic Ocean (NEA), three in the Mediterranean Sea (MED) and two in the south-western Pacific Ocean (SWP). Maximum numbers of alleles per locus ranged from 9 to 38 (sample mean, 5.2–22.6 per locus; overall mean, 14.2 ± 0.47 SE), and observed heterozygosities per locus ranged from 0.44 to 1.00 (overall mean: 0.79 ± 0.19 SE). Significant deficits of heterozygotes were observed in 20% of tests. Multilocus F _ST values were observed ranging from 0.00 to Θ = 0.036 and Θ′ = 0.253, with a mean of Θ = 0.013 and Θ′ = 0.079. Pairwise F _ST values showed that the SWP, NEA and MED stocks were significantly distinct from one another, thus corroborating findings in previous studies based on mitochondrial DNA, nuclear DNA (other than microsatellites) and allozyme analyses. Heterogeneity was observed for the first time between samples within the Mediterranean Sea. GENELAND indicated the potential presence of three populations across the NEA and two separate populations in the Mediterranean Sea. Observed genetic structure may be related to migration patterns and timing of movements of subpopulations to the feeding grounds in either summer or autumn. We suggest that a more intensive survey be conducted throughout the entire fishing season to ratify or refute the currently accepted genetic homogeneity within the NEA albacore stock.     

Sibling species in interstitial flatworms: a case study using<Emphasis Type="Italic"> Monocelis lineata</Emphasis> (Proseriata: Monocelididae)

The genetic relationships between morphologically indistinguishable marine and brackish-water populations of Monocelis lineata (O.F. Müller, 1774) (Proseriata: Monocelididae) were analysed by means of allozyme electrophoresis. Fifteen samples of M. lineata (13 from the Mediterranean and two from the Atlantic) from coastal marine and brackish-water habitats were examined for variation at 18 loci. Eleven loci were polymorphic in at least one population of M. lineata. Low levels of within-population genetic variability were found, with average observed and expected heterozygosity values ranging from H_o=0.015±0.015 to 0.113±0.044, and from H_e=0.028±0.028 to 0.138±0.054, respectively. The occurrence of a number of private alleles indicated a marked genetic divergence among populations of M. lineata, with Rogers genetic distances ranging from D_R=0.003 to 0.676 and a highly significant F_ST value (0.918±0.012, P<0.001). UPGMA (unweighted pair-group method using arithmetic average) cluster analysis and multidimensional scaling showed a clear genetic divergence between marine and brackish-water populations. Moreover, Atlantic and Mediterranean populations were sharply separated. Our results suggest that M. lineata is a complex of sibling species.Communicated by R. Cattaneo-Vietti, Genova     

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     

Comparative population genetic structure of marine gastropods (Littorina spp.) with and without pelagic larval dispersal

 Population genetic theory predicts that marine animal species with planktonic larvae will have less genetic structure than those with direct development. We compared the genetic structure of four species of littorinid snails – two with planktonic egg capsules that hatch as planktonic larvae and two with benthic egg masses that hatch as crawl-away juveniles. We used DNA sequencing and single stranded conformational polymorphism (SSCP) to assess sequence variation in a 480 bp fragment of the mitochondrial cytochrome b gene and then used an analysis of molecular variance (AMOVA) to estimate Φ_st for populations from the northeastern Pacific coast. One of the two direct-developing species, Littorina subrotundata, had a moderate amount of population structure (Φ_st=0.209) as expected but the other direct-developing species, L. sitkana, was nearly fixed for a single haplotype that made it impossible to precisely estimate Φ_st. One of the two planktonic-developing species, L. scutulata, did not show any significant population structure (Φ_st=0.004). In contrast to our expectations, the other planktonic-developing species, L. plena, showed some weak but statistically significant population structure (Φ_st=0.052). We discuss how differences in population genetic structure between species with the same type of development may reflect differences in their historical demography. Received: 22 December 1999 / Accepted: 24 July 2000     

Interactions between NH+4 and NO−3 uptake and assimilation: comparison of diatoms and dinoflagellates at several growth temperatures

Ammonium concentrations of ∼1 M are commonly cited as being the threshold for inhibition of NO₃ ⁻ uptake, but the applicability of this threshold to phytoplankton from different taxonomic classes has rarely been examined. Additionally, little is known about the influence of environmental variables (e.g. growth temperature) on the interaction between ambient NH₄ ⁺ and NO₃ ⁻ uptake. Four species of estuarine phytoplankton, two diatom [Chaetoceros sp., and Thalassiosira weissflogii (Grunow) Fryxell et Hasle] and two dinoflagellate [Prorocentrum minimum (Pavillard) Schiller, and Gyrodinium uncatenum Hulburt], were grown on NO₃ ⁻ at several different temperatures (4, 10, 15, or 20 °C), and the impact of NH₄ ⁺ additions on NO₃ ⁻ uptake/assimilation (non-TCA-extracted) and assimilation (TCA-extracted) was assessed. For all species at all temperatures, NO₃ ⁻ uptake/assimilation and assimilation rates decreased in a roughly exponential manner with increasing NH₄ ⁺ concentrations but were not completely inhibited even at elevated NH₄ ⁺ concentrations of 200 μM. Estimated half-inhibition concentrations (K _i) were significantly greater in the diatom species (mean ± SE; 2.70 ± 0.67 μM) than in the dinoflagellate species (1.26 ± 0.55 μM). Half-inhibition constants were positively related to temperature-limited relative growth rate although not significantly. The observed inhibition of NO₃ ⁻ uptake and assimilation, as a percentage of NO₃ ⁻ uptake in the absence of NH₄ ⁺, averaged about 80% and ranged from 49 to 100%. For all species, a significant (P < 0.001) positive correlation was found between percent inhibition of NO₃ ⁻ assimilation and temperature-limited relative growth rate. Two experiments on Chesapeake Bay phytoplankton during an April 1998 diatom bloom showed that in short-term (∼1 h) temperature manipulation experiments, percent inhibition of NO₃ ⁻ uptake/assimilation was also positively related (P = 0.05) to experimental temperature. The observed relationships between temperature-limited relative growth rate and percent inhibition of NO₃ ⁻ assimilation rates for the species tested suggest that at the enzyme level, the inhibitory mechanism of NO₃ ⁻ assimilation is similar among species, but at the whole cell level may be regulated by species-specific differences in the accumulation of internal metabolites. These findings add not only to our understanding of species-specific variability and the role of growth temperature, but also provide additional data with which to evaluate current models of NH₄ ⁺ and NO₃ ⁻ interactions. Received: 31 August 1998 / Accepted: 7 December 1998     

Ecophysiological constraints associated with aggression,and evolution toward pacifism in Spalax ehrenbergi

Evolution from a solitary way of life to sociality involves changes in the modalities of within-population aggression. This paper is concerned with environmental causes of variation in aggression within the complex of mole-rat species Spalax ehrenbergi. We focus on physiological constraints associated with aggressive behaviour, i.e. those resulting from emotional arousal and thus stress. Aggression, social stress sensitivity and urine osmolarity were studied in males of two chromosomal species of Spalax ehrenbergi, 2n = 52 (occurring in cool humid areas) and 2n = 60 (occurring in warm arid zones). Males with 2n = 52 reached a higher level of within-population aggression during a 3-round tournament (an average, 488 ± 140 acts) than males with 2n = 60 (163 ± 54 acts). The species with 2n = 52 displayed a higher corticosterone response after a social encounter (in average, 5.15 ± 0.54 μg/100 ml) than that with 2n = 60 (3.0 ± 0.3 μg/100 ml) whereas resting levels of this hormone were low and not different between the two species. Urine osmolarity was significantly higher in males with 2n = 60 (331.9 ± 18.6 mmol/kg) than males with 2n = 52 (267.3 ± 10.9 mmol/kg). Considering the overall general trend urine osmolarity tended to be negatively related to the level of aggression, while corticosterone levels tended to vary positively with aggressiveness (Fig. 3). Our results allow further discussion of the hypothesis that high levels of aggression could be selected against in arid habitats due to their physiological correlates (decrease in water economy, increased corticosterone leading to potential energy mobilisation and loss). We propose that behavioural ecophysiology could have contributed to the radiation of the S. ehrenbergi superspecies into increasingly arid environments, and may favour evolution toward social tolerance. Received: 10 June 1995/Accepted after revision: 25 October 1995     

Effect of Ca2+ on survival and development of metamorphosing bonefish (Albula sp.) leptocephali

Bonefish (Albula sp.) larvae (leptocephali) from the Gulf of California complete metamorphosis in ˜10 d in natural seawater (35‰S; Ca²⁺ conc = 10.5 mM). The increase in ossification that occurs near the end of the non-feeding metamorphic period, in addition to the ability of larvae to complete metamorphosis in dilute seawater (8‰ S) prompted the present study, where the effects of varying the external calcium ion concentration, [Ca²⁺]_e, of artificial seawater (ASW) on the survival, development and internal (whole-body) calcium ion content, (Ca²⁺)_i, of unfed metamorphosing larvae were investigated. Early-metamorphosing larvae placed in␣ASW, where [Ca²⁺]_e = 10.1 mM, survived for up to 10 d and developed normally without exogenous nutrients. In shorter-term experiments (4 to 5 d), no differences in survival were found for larvae in ASW with [Ca²⁺]_e rang-ing from 1.5 to 10.1 mM. However, in Ca²⁺-free ASW, most larvae died within 27 h and no larvae survived more than 42 h; the median lethal time (LT₅₀), and its 95% confidence limits, were 14.5 (10.0 to 20.9) h. High mortality (81% after 20 h) also occurred in 1.0 mM Ca²⁺ ASW, but 2 of 16 larvae tested survived for 96 h. The 96 h median tolerance limit (TL_M), corrected for control mortality, was 1.2 mM Ca²⁺. In natural seawater, larval (Ca²⁺)_i remained relatively constant ( = 0.419 mg larva⁻¹)␣in early- and intermediate-metamorphosing larvae, and then increased to a mean value of 0.739 mg larva⁻¹ in advanced larvae, indicating that Ca²⁺ was␣taken up from the medium at this stage; the increase in (Ca²⁺)_i corresponded to the period of ossification of the vertebral column. Internal (whole-body) magnesium ion content (Mg²⁺)_i showed no significant change during metamorphosis ( = 0.089 mg larva⁻¹). No significant differences in (Ca²⁺)_i were found in advanced larvae in natural seawater and those in ASW, with [Ca²⁺]_e ranging from 2.0 to 10.1 mM. However, clearing and staining revealed that ossification of the vertebral column had not yet occurred in advanced larvae from 2.0 to 10.1 mM Ca²⁺ ASW. Also, low [Ca²⁺]_e (1.0 to 2.0 mM) usually produced deformed larvae that swam erratically, at times showing “whirling” behavior. Received: 21 August 1996 / Accepted: 26 August 1996     

Halogenated metabolites in two marine polychaetes and their planktotrophic and lecithotrophic larvae

This study investigated the occurrence and ontogenetic changes of halogenated secondary metabolites in planktotrophic and lecithotrophic larvae and adults of two common, infaunal polychaetes, Streblospio benedicti (Spionidae) and Capitella sp. I (Capitellidae), with different life-history traits. S. benedicti contains at least 11 chlorinated and brominated hydrocarbons (alkyl halides) while Capitella sp. I contains 3 brominated aromatic compounds. These halogenated metabolites are potential defense compounds benefiting both larvae and adults. We hypothesized that: (1) planktotrophic larvae contain halogenated metabolites because they are not protected by adult defenses, (2) quantitative and qualitative variation of planktotrophic larval halogenated metabolites parallels that of adults, and (3) brooded lecithotrophic larvae initiate the production of halogenated metabolites only after metamorphosis. To address these hypotheses, volatile halogenated compounds from polychaete extracts were separated using capillary gas chromatography and identified and quantified using mass spectrometry with selected ion monitoring. All four life stages (pre- and post-release larvae, new recruits, adults) of both S. benedicti and Capitella sp. I contained the halogenated metabolites previously identified from adults. This is the first report of halocompounds identified and quantified in polychaete larvae. Allocation of potential defense compounds to offspring varied as a function of species, feeding type and developmental stage. Pre-release larvae of S. benedicti with planktotrophic development contained the lowest concentration of total halogenated metabolites (1.75 ± 0.65 ng mm⁻³), post-release and new recruits contained intermediate concentrations (8.29 ± 1.72 and 4.73 ± 2.63 ng mm⁻³, respectively), and planktotrophic adults contained significantly greater amounts (28.9 ± 9.7 ng mm⁻³). This pattern of increasing concentrations with increasing stage of development suggests synthesis of metabolites during development. Lecithotrophic S. benedicti post-release larvae contained the greatest concentrations of halometabolites (71.1 ± 10.6 ng mm⁻³) of all S. benedicti life stages and developmental types examined, while the amount was significantly lower in new recruits (34.0 ± 15.4 ng mm⁻³). This pattern is consistent with a previously proposed hypothesis suggesting a strategy of reducing potential autotoxicity during developmental transitions. Pre-release lecithotrophic larvae of Capitella sp. I contained the highest concentration of total halogenated metabolites (1150 ± 681 ng mm⁻³), whereas the adults contained significantly lower total amounts (126 ± 68 ng mm⁻³). All concentrations of these haloaromatics are above those known to deter predation in previously conducted laboratory and field trials. As a means of conferring higher larval survivorship, lecithotrophic females of both species examined may be expending more energy on chemical defenses than their planktotrophic counterparts by supplying their lecithotrophic embryos with more of these compounds, their precursors, or with energy for their synthesis. This strategy appears common among marine lecithotrophic larval forms. Received: 14 July 1999 / Accepted: 20 January 2000     

Evidence of limited gene flow in three species of coral reef fishes in the lagoon of New Caledonia

Allozyme variation was used to investigate the genetic structure of the coral reef fishes Stegastes nigricans, Epinephelus merra and Acanthurus triostegus around New Caledonia. Each species was sampled from each of three sites in the ≃1000 km circumference of the lagoon of New Caledonia. Allelic variation was recorded for each species at 14, 13 and 17 loci, respectively, and heterozygosity diversity (H s) was 0.082, 0.065 and 0.116, respectively. Analysis of genetic differentiation between sites produced inconsistent results between species, with spatial heterogeneity in two species (S. nigricans, F st  = 0.038; A. triostegus, F st = 0.049) and homogeneity in one species (E. merra, F st = 0.000). Hydrological and climatic data from the lagoon suggest that the eastern and western sides of the lagoon are isolated, since they lie in water masses of different origin. This may explain the genetic differentiation and restricted gene flow found at a local scale for S. nigricans and A. triostegus. Homogeneity in populations of E. merra is discussed in relation to its low genetic diversity and its reproductive behaviour. Received: 23 April 1997 / Accepted: 25 September 1997