Flowering of the seagrass Posidonia oceanica in a north-western Mediterranean coastal area: temporal and spatial variations

Sexual reproduction in Posidonia oceanica (L.) Delile is likely to be highly variable both in space and time. However, there has been little effort to identify at what spatial scale variability exists in the distribution and abundance of reproductive shoots or the magnitude of these variations. Such information is important for elucidating several aspects of the reproductive strategy of this species. Here, by using a reconstructing technique, I estimated the rate at which flowering events occurred over the last decades in three P. oceanica meadows along the coastline of the north-western Mediterranean (Italy). Then, variation in inflorescence frequency (i.e. the percentage of shoots that flowered in a particular year of the total number of shoots present in the sample in that year) was examined on a hierarchy of spatial scales, ranging from a few metres to meadows separated by tens of kilometres over a 3-year period. Separate analyses were also conducted to examine the patterns of spatial variability of several plant characteristics (i.e. shoot age and density of vegetative shoots) that may influence the timing of flowering. In total, ten reconstructed flowering events were recorded during the period 1983–1999. Despite the similar climatic conditions experienced in the three meadows surveyed, no inflorescence scars were found in one meadow. In the other two meadows, the flowering rate fluctuated considerably from year to year (0–26%), and simultaneous flowerings were detected only in 3 years (1993, 1994 and 1995). The pattern of spatial variability in the inflorescence frequency varied among years. An investigation of the flowering events recorded along the north-western Mediterranean coast reveals that in the studied period flowering also occurred across a broad range of meadows. Therefore, flowering could be a far more widespread phenomenon in this sector of the Mediterranean basin than previously thought. Analysis of variance showed significant variation in 1993 at the mean inflorescence frequency on very small scales (metres to tens of metres), whereas variation was not found at any of the scales investigated in 1994 and 1995. Overall, variation in the inflorescence frequency within individual meadows (65–93%) was at least an order of magnitude greater than that among separate meadows (0.5–19%). Patterns of spatial variation in the plant characteristics examined did not reflect those of flowering. It may be hypothesised that internal rhythms of populations, in conjunction with changes in environmental conditions, which are consistent over relatively large spatial (kilometres), are involved in the control of flowering induction. Environmental heterogeneity (in terms of nutrient availability) on a very local scale (metres to tens of metres) and intrinsic factors might affect the distribution of reproductive shoots within P. oceanica meadows.Communicated by R. Cattaneo-Vietti, Genova     

A spatial gradient in the potential reproductive output of the sea mussel <Emphasis Type="Italic">Mytilus californianus</Emphasis>

Spatial variation in reproductive output from different populations within a region could have important consequences for recruitment, and cascading effects on populations and communities of marine species, but is rarely examined over meso-scales (i.e., tens to hundreds of kilometers). In this study, reproduction in the dominant mid-intertidal mussel, Mytilus californianus, was examined from sites spanning Point Conception, California over a 6-month period (March–August 2000). There was a dramatic geographic pattern in the relationship between size and potential reproductive output that was qualitatively similar across all 6 months sampled. Increases in allocation to reproductive tissue with increasing body size occurred at all sites, but the slope nearly doubled at sites south of Point Conception compared to northern sites. The spatial variation in size-specific reproductive output, coupled with additional spatial gradients in mussel density and size distributions, combined to increase total reproductive output by over eightfold at southern relative to northern sites. This study highlights the need to explicitly examine spatial patterns of reproductive output at these meso-scales, in order to better understand connectivity and source–sink dynamics in marine systems.     

Spatial patterns of spawning in the coral-reef damselfish Dascyllus albisella

Reproductive patterns of the endemic Hawaiian damselfish Dascyllus albisella were examined around the island of Oahu for 5 m in 1992. Daily variation in spawning was studied at four spatial scales: individual territories; reef transects; whole patch reefs; and locations separated by>20 km. A 6 d, non-lunar nesting cycle was found for individual males, and a synchronized, 6 d, nonhinar spawning cycle was found within reef transects, whole patch reefs, and locations. Spawning synchronicity was maintained among reefs within a location. However, spawning among locations was found to be asynchronous, limiting the spatial scale of reproductive synchrony to>1 km but<20 km. Inherent benefits of synchronous reproduction are discussed for D. albisella, but only the localized swamping of planktonic predators may be an inherent benefit of synchronous spawning for this species. Alternatively, synchronous spawning among populations may exist from adults tracking local, favorable environmental conditions rather than having evolved towards an inherent benefit of synchronization.     

Mode of reproduction,recruitment, and genetic subdivision in the brooding sponge<Emphasis Type="Italic"> Haliclona</Emphasis> sp.

Sponges display a variety of reproductive strategies that have the potential to influence population genetic structure. Histological examination of ten reproductive individuals of the Western Australian sponge Haliclona sp. showed that this species broods embryonic larvae that are potentially limited in dispersal capabilities. Because sponges have the potential to propagate in a number of modes, allozyme electrophoresis was used to assess the relative importance of asexual and sexual reproduction to recruitment, and to quantify genetic subdivision over different spatial scales. Tissue samples from 227 sponges were collected from reefs within two areas 400 km apart: Hamelin Bay and Rottnest Island. Contrary to expectations for highly clonal populations, genotypic diversity within sites was high, no linkage disequilibrium was found, and there was no evidence of genotypic clustering within reefs. There was no genetic evidence that asexual reproduction is important for the maintenance of populations. Genetic comparisons were consistent with mixing of sexually produced recruits within reefs, on a scale up to a few hundred metres, but significant genetic subdivision between reefs (F_ST=0.069 at Hamelin Bay, 0.130 at Rottnest Island) indicated that water gaps of several hundred metres are effective at preventing dispersal. Subdivision between the two areas, separated by 400 km, was moderately greater (F_ST=0.142) than within, but the same alleles were predominant in the two areas. These genetic patterns are consistent with limited dispersal capabilities of brooded larvae.Communicated by G.F. Humphrey, Sydney     

Reproductive synchrony in a habitat-forming kelp and its relationship with environmental conditions

Key biological processes such as the timing and synchrony of reproduction, are sensitive to fluctuations in the environment, as well as alterations in the physiology and behaviour of individuals and the ecology of populations. The main aim of this study was to identify patterns in reproductive timing and relationships with prevailing environmental conditions, for the major habitat-forming alga in temperate Australia, Ecklonia radiata (C. Agardh) J. Agardh. We observed strong synchronous patterns of zoospore production across spatial scales from hundreds of metres to tens of kilometres, with populations of E. radiata reproducing from mid-summer to the end of autumn. High zoospore densities were positively related to sea temperature and E. radiata released zoospores in temperatures well above previously documented thermal tolerance limits for other Laminariales. Reproductive timing was also negatively related to seasonal increase in wave exposure. We conclude that E. radiata undergoes synchronous reproduction which can be linked to its annual growth cycle and natural environmental fluctuations, to promote growth and survival of recruits.     

Non-random mating and population genetic subdivision of two broadcasting corals at Ningaloo Reef,Western Australia

Allozyme electrophoresis of two corals was used to assess whether populations at Ningaloo Reef, Western Australia are primarily self-seeding or whether recruitment is from a broader geographic pool. Significant genetic subdivision across a range of spatial scales (between 6.5 km and 155 km) was found for both Acropora digitifera and A. aspera, with mean F _ST values of 0.010 and 0.067 respectively. Large departures from Hardy-Weinberg expectations were found for both species. Without exception these were due to deficits of heterozygotes; mean D values were –0.341 for A. digitifera and –0.455 for A. aspera. The magnitude of the deficits was consistent both across loci for all sites and across all sites for each locus. Some loci were found to be in linkage disequilibrium but no consistent pattern was observed. Also, multi-locus genotypic diversity values were generally high (between 0.83 and 1.00) and so departures from equilibria cannot be attributed to asexual reproduction. The most plausible explanation for the patterns observed is restricted gene flow at both the planktonic and gametic stages, with mating between close relatives.Communicated by G.F. Humphrey, Sydney     

Evidence for restricted gene flow over small spatial scales in a marine snapping shrimp <Emphasis Type="Italic">Alpheus angulosus</Emphasis>

Despite the apparent absence of geographic barriers, connectivity among marine populations may be restricted by, for example, ecological or behavioral mechanisms. In such cases, populations may show genetic differentiation even over relatively small spatial scales. Here, mitochondrial sequence data from the cytochrome oxidase I (COI) gene and seven polymorphic microsatellite markers were used to investigate fine geographic scale population genetic structure in the snapping shrimp Alpheus angulosus, a member of the A. armillatus species complex, from collections in Florida, Jamaica, and Puerto Rico carried out from 1999 to 2005. The COI data showed a deep divergence that separated these samples into two mitochondrial clades, but this divergence was not supported by the microsatellite data. The COI data reflect past population divergence not reflected in extant population structure on the whole genome level. The microsatellite data also revealed evidence for moderate population structure between populations as close as ∼10 km, and no evidence for isolation by distance, as divergences between near populations were at least as strong as those between more broadly separated populations. Overall, these data suggest a role for restricted gene flow between populations, though the mechanisms that reduce gene flow in this taxon remain unknown.     

Genetic differentiation among populations of the Mediterranean hermaphroditic brooding coral<Emphasis Type="Italic"> Balanophyllia europaea</Emphasis> (Scleractinia: Dendrophylliidae)

Spatial models of genetic structure and potential gene flow were determined for five populations of Balanophyllia europaea, a simultaneous hermaphroditic and brooding coral, endemic to the Mediterranean. Six allozyme loci indicated a genetic structure that departed markedly from Hardy–Weinberg equilibrium, with a significant lack of heterozygotes. The genetic structure observed supports the hypothesis that self-fertilisation characterises the reproductive biology of B. europaea. Populations at small spatial scales (8–40 m) are genetically connected, while those at large scales (36–1,941 km) are genetically fragmented; the genetic differentiation of the populations is not correlated to geographic separation. This spatial model of genetic structure is compatible with an inbreeding mating system. Furthermore, it is also consistent with the expected dispersal potential of swimming larvae of brooding corals, i.e. larvae that are able to produce significant gene flows only within limited spatial scales.Communicated by R. Cattaneo-Vietti, Genova     

Scales of variation in otolith elemental chemistry of juvenile staghorn sculpin (<Emphasis Type="Italic">Leptocottus armatus</Emphasis>) in three Pacific Northwest estuaries

Although distinct otolith elemental signatures are often observed in fish collected from different estuaries, significant differences are also observed among sites within estuaries. Variation at these smaller spatial scales is not well quantified and has the potential to lead to inappropriate interpretations of otolith elemental data. To quantify variation at multiple scales, the otolith elemental composition (Mg:Ca, Mn:Ca, Sr:Ca, Ba:Ca, and Pb:Ca) of juvenile staghorn sculpin (Leptocottus armatus Girard, 1854) collected from five sites within three estuaries, the Columbia River (two sites) and Coos Bay (one site), Oregon, and Humboldt Bay, California (two sites), was examined. Using laser ablation-inductively coupled plasma mass spectrometry, each otolith was sampled at three zones: (1) within the primordium, which represents the egg and early larval periods; (2) at the outer edge, which represents the juvenile period just prior to collection; and (3) midway between the primordial and edge samples, which represents the late larval and early juvenile period. There were significant differences in otolith metal-to-calcium ratios at all scales examined. Using multi-element otolith signatures, fish were classified to estuary and site within estuary with relatively high levels of accuracy (av = 70–90%). The largest differences in metal-to-calcium ratios were observed between sites within estuaries (<5 km apart) and the smallest differences were observed among otolith zones. Variation in otolith chemistry may be used to provide information on probable habitat use by estuarine fish but studies must be carefully designed. Electronic supplementary material  Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Environmental determinants of distribution and foraging behaviour of cormorants (Phalacrocorax spp.) in temperate estuarine habitats

The distribution and behaviour of cormorants in estuarine environments was examined on the central coast of New South Wales, Australia, with respect to habitat associations at different spatial scales. No consistent variation in abundance was found for four species of cormorants (great Phalacrocorax carbo, pied P. varius, little black P. sulcirostris, and little pied P. melanoleucos) with state of tide (high and low) and time of day (early, middle, and late) at five estuarine locations. Differences in abundance were found among locations that were not confounded by short-term temporal variation (i.e. time of day and tide). Differences in abundance were detected among habitats (e.g. bays, creeks, and headlands) separated by hundreds of metres to kilometres in different estuaries. Cormorants of all species were rare on the open coast and near the entrance of estuaries. Abundances of cormorants varied greatly within and among creeks, bays, and river channels. Presence of seagrass beds explained much of this variation and most of the cormorants swimming and feeding were found near seagrass. Mapping of seagrass beds and the positions of cormorants at scales of metres to hundreds of metres showed a close relationship between the presence of swimming and roosting beds and the presence of seagrass beds for P. melanoleucos and for P. carbo. We argue that cormorants make decisions to visit particular estuarine habitats, especially those with seagrass, where many types of prey (e.g. fish and crustaceans) are probably most abundant. These choices must be interpreted in the context of decisions that cormorants make on scales of hundreds to thousands of kilometres during periodic excursions to the interior of Australia. Further, environmental threats to seagrass beds could impinge on these mobile visitors to the same extent as on more permanent residents. Received: 14 February 2000 / Accepted: 17 July 2000     

Accumulation of cadmium from contaminated water and sediment by the shrimp Callianassa australiensis

The burrowing marine shrimp Callianassa australiensis (Dana) was collected from an uncontaminated area in Western Port, Victoria, Australia in 1977. The shrimp were exposed to cadmium-contaminated water and sediment for 8 wk. The concentrations ranged from 0.5 to 63 g Cd 1^-1 for water and 0.5 to 63 g Cd g^-1 for sediment. The shrimp accumulated cadmium from water at a rate commensurate with increases in the concentration of cadmium in water and the duration of the experiment. Although the cadmium concentration in the sediments was 10³ times higher than that in water, it hat no effect on cadmium uptake by the shrimp. The concentration factors decreased with increasing concentration of cadmium in water but increased as the duration of exposure increased. The shrimp dry weight decreased with increasing concentration of cadmium in water and duration of exposure. As was the case with cadmium uptake by the shrimp, these two factors acted interactively on the shrimp dry weight, but the third factor, concentration of cadmium in sediment, had no effect.     

Isolation by distance in the scleractinian coral <Emphasis Type="Italic">Seriatopora hystrix</Emphasis> from the Red Sea

Pelagic dispersal of larvae in sessile marine invertebrates could in principle lead to a homogeneous gene pool over vast distances, yet there is increasing evidence of surprisingly high levels of genetic differentiation on small spatial scale. To evaluate whether larval dispersal is spatially limited and correlated with distance, we conducted a study on the widely distributed, viviparous reef coral Seriatopora hystrix from the Red Sea where we investigated ten populations separated between ~0.150 km and ~610 km. We addressed these questions with newly developed, highly variable microsatellite markers. We detected moderate genetic differentiation among populations based on both F _ST and R _ST (0.089 vs. 0.136, respectively) as well as considerable heterozygote deficits. Mantel tests revealed isolation by distance effects on a small geographic scale (≤20 km), indicating limited dispersal of larvae. Our data did not reveal any evidence against strictly sexual reproduction among the studied populations.     

Time and space: genetic structure of the cohorts of the common sea urchin <Emphasis Type="Italic">Paracentrotus</Emphasis><Emphasis Type="Italic">lividus</Emphasis> in Western Mediterranean

Spatio-temporal variability in settlement and recruitment, high mortality during the first life-history stages, and selection may determine the genetic structure of cohorts of long-lived marine invertebrates at small scales. We conducted a spatial and temporal analysis of the common Mediterranean Sea urchin Paracentrotus lividus to determine the genetic structure of cohorts at different scales. In Tossa de Mar (NW Mediterranean), recruitment was followed over 5 consecutive springs (2006–2010). In spring 2008, recruits and two-year-old individuals were collected at 6 locations along East and South Iberian coasts separated from 200 to over 1,100 km. All cohorts presented a high genetic diversity based on a fragment of mtCOI. Our results showed a marked genetic homogeneity in the temporal monitoring and a low degree of spatial structure in 2006. In 2008, coupled with an abnormality in the usual circulation patterns in the area, the genetic structure of the southern populations studied changed markedly, with arrival of many private haplotypes. This fact highlights the importance of point events in renewing the genetic makeup of populations, which can only be detected through analysis of the cohort structure coupling temporal and spatial perspectives.     

Genetic structure of the subtidal red alga Delisea pulchra

We used random amplified polymorphic DNA (RAPDs) to examine small-scale spatial genetic structure in the red alga Delisea pulchra (Greville) Montagne at two locations near Sydney, Australia. We examined genetic structure among plants at four spatial scales ranging from 2 km apart down to <50 cm apart between locations, among sites within locations, among quadrats within sites, and among plants within quadrats. Haploid stages of D. pulchra were absent from the populations studied, suggesting that they are maintained through asexual reproduction of diploid plants. Consistent with this, we found that 19 RAPD phenotypes scored in this study had multiple individuals, indicating the presence of clones in these populations. However, there were no RAPD phenotypes common to two locations separated by only 2 km. Analysis of molecular variance revealed that strong genetic differences occurred between plants from these two locations, with 46.3% of the total genetic variation occurring at this scale, most probably reflecting limited gene flow. Within each location, <25% of the genetic variation was attributable to differences among sites or quadrats, indicating gene flow at those smaller scales. Most of the variation within each location occurred at the smallest spatial scale, among plants within 0.25 m² quadrats. Nonetheless, some pairwise genetic distances (φST) between sites or quadrats within locations were large, indicating some genetic divergence on smaller scales. Genetic distance was independent of spatial distance within both locations, suggesting that fine-scale differences within locations were most probably caused by variation in fine-scale patterns of water movement or fine-scale natural selection. We assessed the impact of one potential selective agent, grazing sea urchins, on the fine-scale genetic structure of D. pulchra. There was no evidence that grazing by sea urchins affected the genetic structure of D. pulchra. In combination with demographic data, our results indicated that local populations of D. pulchra within locations were relatively open and that fine-scale genetic structure was probably constrained by gene flow. At the larger scale however, strong genetic differentiation indicated little gene flow between locations and restricted dispersal of spores. Received: 22 April 1999 / Accepted: 29 November 1999     

Population connectivity in the temperate damselfish <Emphasis Type="Italic">Parma microlepis</Emphasis>: analyses of genetic structure across multiple spatial scales

This study investigated the utility of microsatellite markers for providing information on levels of population connectivity for a low dispersing reef fish in New South Wales (NSW), Australia, at scales ≤400 km. It was hypothesized that the temperate damselfish Parma microlepis, which produces benthic eggs and has limited post-settlement dispersal, would exhibit spatial genetic structure and a significant pattern of isolation-by-distance (IBD). A fully nested hierarchical sampling design incorporating three spatial scales (sites, location and regions, separated by 1–2, 10–50 and 70–80 km respectively) was used to determine genetic variability at seven microsatellite loci. Broad-scale genetic homogeneity and lack of IBD was well supported by single and multi-locus analyses. The proportion of the total genetic variation attributable to differences among regions, locations or sites was effectively zero (Φ/R-statistics ≤0.007). The geographic distribution of genetic diversity and levels of polymorphism (H _E 0.21–0.95) indicate high mutation rates, large effective population sizes, and high rates of gene flow. Significant gene flow may be driven by factors influencing pre-settlement dispersal, including the East Australian Current (EAC) and habitat continuity. Genetic connectivity may not reflect demographically important connectivity, but does imply that P. microlepis populations are well connected from an evolutionary perspective. Total observed genetic diversity was accounted for within 1–2 km of reef and could be represented within small Marine Protected Areas. Reef fishes in NSW which have life histories similar to P. microlepis (e.g. pre-settlement durations ≥2 weeks) are also likely to exhibit genetic homogeneity. Genetic markers are, therefore, most likely to provide information on demographically relevant connectivity for species with lower dispersal capabilities, small population sizes, short life spans, and whose habitats are rare, or patchily distributed along-shore. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Ecological genetics of the barnacle,<Emphasis Type="Italic"> Hexaminius foliorum</Emphasis>, within mangrove forests near Sydney,Australia

Spatial and temporal distribution of allozyme variation at three loci in a cohort of the barnacle, Hexaminius foliorum, living on leaves of Avicennia marina was studied from recruitment to adulthood at three geographical scales. Analysis of populations shortly after recruitment showed that there were significant divergences in allele frequencies at the coarsest geographical scale studied (between estuaries, 50 km apart, Wright's F statistic=0.016) and at the finest geographic scale (between sites, 50–100 m apart, Wright's F statistic=0.018). There was, however, no significant genetic divergence at an intermediate scale (between bays, 3–4 km apart, Wright's F statistic=0.002). The genetic differences between populations decreased over time due to the selection against the null homozygotes originally present at high frequency at two loci. There was sufficient mortality (ranging from 35.5 to 80%) between seasons to account for the deaths needed for the observed changes in allele frequencies. Differences in the genetic structure between estuaries may be the result of isolation and limited mixing of cyprid larvae among estuaries. Differences in the genetic structure between sites may be due to pre- and post-settlement mortality acting on H. foliorum.Communicated by G.F. Humphrey, Sydney     

Variation at local scales need not impede tests for broader scale patterns

Ecologists are not always mindful of the constraints imposed by their scale of observation and sometimes prematurely attempt broad generalisations or become mesmerised by local details depending on their predilections. We specifically chose a character that is known for its local and unpredictable variation (morphology of kelp) to test the effect of scale on our ability to determine spatial patterns. We compared the morphology of Ecklonia radiata between monospecific and mixed stands of canopy-forming algae across temperate Australia (>5,100 km coastal distance) within a hierarchy of several spatial scales. While E. radiata specimens were generally larger in monospecific than in mixed stands, we failed to observe differences in morphology between stands at many sites and locations. Despite substantial local variation, differences between stands became increasingly clear at broader scales. The frequency of inconsistent differences between stands was greatest at local scales (sites separated by kms), intermediate at intermediate scales (locations separated by 100s of kms) and least at regional scales (regions separated by 1,000s of kms). These observations support the idea that large scale patterns can emerge from apparent stochasticity at small scales, and that unaccountable variation at local scales need not impede tests for similar patterns at broader scales. Most ecologists work at scales where complexity tends to be greatest (i.e. local) and is likely to be explained by special and unique events. It is encouraging, therefore, to observe that patterns can emerge from complexity at local scales to provide new opportunities to answer some of the more interesting questions about the relative importance of processes across the vast parts of the worlds coast.Communicated by M.S. Johnson, Crawley     

Supply-side ecology of the brown mussel,<Emphasis Type="Italic"> Perna perna</Emphasis>: an investigation of spatial and temporal variation in,and coupling between,gamete release and larval supply

Sampling of recruitment-associated variables of Perna perna was done approximately monthly for 14 months at intertidal locations 500 m apart, nested within sites 25 km apart. Paired with intertidal locations were nearshore locations, 600 m to sea. Sampling assessed spawning, densities of larvae in the water column and densities of late plantigrades and juveniles on the shore. Major events in each variable were synchronous over larger scales (10s of kilometres) while subsidiary events were synchronised at smaller scales, varying within sites (100s of metres) or even within locations (metres). This suggests that the processes driving major events operated over large scales while processes operating at much more local scales drove less intense, more localised events. A major spawning event occurred at all locations in May–June 1998. Weaker spawning events occurred at different times in different locations. Larvae were found on 80% of sampling occasions, densities peaking in January–March 1998 and 1999 at all locations. Plantigrades and juveniles showed less clear patterns, with considerable residual variation. There was no sign of strong coupling among variables with few significant direct or cross correlations. The major sources of variability shifted from time to space as one progressed from spawning, to plantigrade density to juvenile density. For spawning, time was the most important source (58%) of heterogeneity and space accounted for little (8%) of the total variance. For larvae and late plantigrades, time was still the most important source of variability (41% and 33%, respectively), but space was a much more substantial component. For juveniles, small-scale (residual) spatial variability dominated total variability (75%). This strongly suggests the importance of hydrography and its effects on variation in delivery of larvae to the intertidal from offshore. These findings also indicate greater spatial heterogeneity as recruits age, reflecting small-scale variations in larval delivery and the increasing importance of post-settlement mortality.Communicated by G.F. Humphrey, Sydney     

Comparisons of genotype-tolerance responses in populations of <Emphasis Type="Italic">Hediste diversicolor</Emphasis> (Polychaeta: Nereididae) exposed to copper stress

The analysis of genotypic shifts in field-exposed populations has been proposed as a tool for monitoring the environmental impacts of contaminants. Previous laboratory experiments showed the occurrence of genotypic shifts induced by copper exposure in samples of Hediste diversicolor from the Pialassa estuary (North Adriatic Sea, Italy). In order to test if genotype-tolerance responses observed were consistent at larger spatial scales, populations of H. diversicolor were sampled in three haphazardly chosen estuaries along the Italian coast (Comacchio, Pialassa, Serchio) and exposed to 0.48 mg l⁻¹ Cu²⁺ under laboratory conditions. Survival analysis procedures were used to test for possible relationships between time-to-death and genotypes at three allozyme loci (ALD, FH, PGI). Genotype-tolerance responses observed at locus ALD were consistent among populations up to 2500 km distant, with genotype ALD ^102/102 associated with the shortest survival times. Comparisons with previous laboratory experiments showed that responses to copper stress at locus ALD were also consistent in time, with genotypes ALD ^102/102 and ALD ^100/102 associated with higher 96 h mortality ratios. Results suggested the occurrence of stable relationships between genotypes at locus ALD and tolerance responses of H. diversicolor to copper stress. Conversely, genotype-tolerance responses at loci PGI and FH were not consistent either in space (among populations from the three estuaries) or in time (between laboratory experiments), being possibly affected by site-specific features of populations or stochastic factors. Results show that genotype-tolerance responses should be carefully verified at different spatial and temporal scales, before considering genotypic shifts among the possible tools for monitoring the effects of environmental stressors.