Recruitment, early survival and growth of the Mediterranean red coral Corallium rubrum (L 1758), a 4-year study

Recruitment, early survival and growth rates of the precious Mediterranean red coral Corallium rubrum (L 1758) are poorly known. We examined these life history traits by means of artificial long-term settlement plates. Eighteen marble tiles placed off the coast of Leghorn (Tuscany, Italy) at two depths (25 and 35 m) were photographed monthly over the 4-year period from 1998 to 2002. Overall, 864 transparencies were examined to follow the individual life histories of red coral colonies belonging to four successive cohorts. Red coral planulae settled on tiles each year between July and September. Overall, 388 settlers colonized the tiles (244 at 25 m and 144 at 35 m), and their respective densities varied between 12.37±6.1 and 2.75±2.4 dm⁻². Heavy mortality affected these colonies (−24.35±9.12 colonies % y⁻¹), but, after 4 years, the tiles still harboured a persistent population (19±4.97 and 9.75±2.87 colonies dm⁻², respectively, at 25 and 35 m) with positive net recruitment rates. Only in 1999 did the net recruitment rate show a negative trend, although only at the shallower depth. At the same time (late summer 1999), a thermal anomaly affected several epibenthic communities in the Ligurian Sea. After 4 years, the tiles were removed, and the colonies that settled on them were measured. The average annual growth rate of colonies was low (0.62±0.19 mm y⁻¹ in diameter), and a marked reduction in growth with age was observed. Our findings suggest that the populations of this slow-growing long-lived octocoral exhibit a high capacity for colonization and seem to be quite resilient to environmental variability.     

A molecular tool for genetic surveys in the red coral (Corallium rubrum): An Inter-Simple Sequence Repeats (ISSRs) perspective

Corallium rubrum is a colonial cnidarian, distributed in Mediterranean and Atlantic rocky bottoms. The species has been largely exploited for use in jewellery since 17th century, raising increasing concerns for the sustainability of the present rate of harvesting. The aim of this study was to standardise a simple and not expensive molecular technique to perform genetic analysis on populations of C. rubrum, in order to provide a useful tool for future surveys. Twelve samples collected from one shallow-water and two deep sites located at the Capo Caccia–Isola Piana MPA (North-western Sardinia, Italy) were surveyed by means of 10 ISSR (Inter-Simple Sequence Repeat) primers. Our results confirmed the ease of use and the reliability of ISSR markers, and their usefulness in detecting genetic variability even among individuals.     

A new method for measuring growth and age in the precious red coral<Emphasis Type="Italic"> Corallium rubrum</Emphasis> (L.)

A new technique for aging red coral Corallium rubrum (L.) colonies based on staining the organic matrix found in the axial calcareous skeleton is presented and validated. This method provided clear-cut images of growth rings of red coral. To demonstrate their annual periodicity, two approaches have been used: (i) applying this technique to adult colonies of known age (more than 20 years old), and (ii) labeling colonies with calcein and allowing them to grow for 1 year. Results provided evidence of the annual periodicity of growth rings. This new method assesses colony age with an underestimate of true age by 3–4 years. The difference between estimated age and actual age could be attributed to the phase of initial growth during which rings are not formed. Colonies from different sites, depths, and habitats (n=33) were analyzed yielding preliminary data on longevity and mean growth rates in red coral. Colonies from shallow habitats (15–62 m) examined here with basal diameter of about 7 mm were at least 30–40 years old. Mean growth rate of basal diameter was 0.35±0.15 mm year^–1. Comparison with previous aging methods used for red coral resulted in considerable differences in estimations of age (about 10 years greater in this study) and growth rates (about four times lower). The application of this method to red coral will provide key data as a basis for developing management and conservation plans for this valuable species.Communicated by: H.R. Lasker     

Coexistence of Abies alba (Mill.) - Fagus sylvatica (L.) and climate change impact in the Iberian Peninsula: A climatic-niche perspective approach

Abies alba Mill. (European silver fir) and Fagus sylvatica L. (beech) are Eurosiberian species dispersed over the Iberian Peninsula. Climate change predictions indicate a rise in temperature and a decrease in precipitation in this region, threatening the future existence of these species. In the present study we analyzed the future topo-climatic suitability of Abies alba and Fagus sylvatica and the mixed forests of these two species, using the General Linear Models technique and data from the third National Forest Inventory (Ministerio de Agricultura PyA, 2007). We considered two modeling approaches based on niche theory: modeling community (Abieti-Fagetum) and overlapping individual species models. General trends showed an overall decrease in both species’ topo-climatic suitability and indicated that the Pyrenees will play a crucial role as a climatic refuge. The modeling approaches markedly differed, however, in their current and future spatial agreement. Despite good accuracy results, community modeling through co-occurrence does not encompass the environmental space of individual species prejudicing future assessments in new environmental situations, suggesting a need for future studies in community modeling.     

Phenology of the Mediterranean seagrass Posidonia oceanica (L.) Delile: Medium and long-term cycles and climate inferences

The results of 15 years of monitoring of Posidonia oceanica in the “Cinque Terre” Marine Protected Area (NW Mediterranean) are presented. Seasonal data on meadow characteristics (cover and shoot density), plant phenology (leaf number, leaf length and width, leaf brown portion, undamaged leaves), lepidochronology, leaf epiphyte cover and herbivore pressure collected from three stations at 5, 10 and 17 m depth were compared. Time-series analyses showed both medium-term (5 < years) and long-term cycles (from 5 to more than 20 years). The comparison of annual cycles with sea surface temperatures (SST) and rainfall showed correlations that differed in relation to depth and, in the case of epiphytes, with each side (internal and external) of the leaf blade. Meadow parameters (visual cover, shoot percent cover) and plant parameters (leaf number, number of undamaged leaves, number of scales shoot⁻¹) showed a positive trend in accordance with the rise of air and sea surface temperature recorded over these last decades. Shoot density and leaf width showed exceptions. Leaf length, leaf brown portion length and the number of undamaged leaves shoot⁻¹ showed positive or negative long-term trends, whose variability could not be related to climate data alone. The two major groups of epiphytes (encrusting algae and the bryozoan Electra posidoniae) showed negative trends. Grazing variability could be explained only partially by climate parameters. Epiphyte cover was found to be related to the NAO index.In conclusion, data showed that the effects of the climate change in terms of both sea surface temperature rising and rainfall decreasing may affect the growth cycles of P. oceanica on two levels: on a decadal level, with positive or negative trends in meadow and plant characteristics and in epiphyte cover; on yearly and seasonal levels, influencing endogenous plant growth rhythms, as in the case of leaf production cycle.     

Genetic structure of Phlebotomus (Larroussius) ariasi populations, the vector of Leishmania infantum in the western Mediterranean: Epidemiological implications

In recent years there has been growing interest in analyzing the geographical variations between populations of different Phlebotomus spp. by comparing the sequences of various genes. However, little is known about the genetic structure of Phlebotomus ariasi. In this study, we were able to sequence a fragment of the mitochondrial Cyt b gene in 133 sandflies morphologically identified as P. ariasi and proceeding from a wide geographical range covering 35 locations in 11 different regions from five countries. The intra-specific diversity of P. ariasi is high, with 45 haplotypes differing from each other by one to 26 bases and they are distributed in two mitochondrial lineages, one limited geographically to Algeria and the other widely dispersed across Mediterranean countries. The Algerian lineage is characterized by having 13 fixed polymorphisms and is made up of one sole haplotype. The European/Moroccan P. ariasi lineage is characterized by being made up of a great diversity of haplotypes (44) which display some geographical structuring. This could be one of the multiple factors involved in the epidemiological heterogeneity of the foci of leishmaniasis. Phlebotomus chadlii is the sister group of European/Moroccan P. ariasi. The separation of the Algerian haplotype, H45, from the rest of the specimens, European/Moroccan P. ariasi and P. chadlii, is well supported by the bootstrap analysis.     

Assessing effects of fishing prohibition on Posidonia oceanica seagrass meadows in the Marine Natural Reserve of Cerbère-Banyuls

The potential of alleviated fishing pressure measures established at the Marine Natural Reserve of Cerbère-Banyuls to affect phenological characters of P. oceanica, the main canopy-forming seagrass, was studied. Our results show differences in some leaf parameters between meadows under fishing prohibition measures compared to those without. In addition shallow P. oceanica meadows inside MPAs had lower non-structural carbohydrate content, yet are able to maintain themselves and, moreover, with an increased density. On the other hand, sexual reproduction was reduced inside protected areas.     

Effects of salinity on leaf growth and survival of the Mediterranean seagrass Posidonia oceanica (L.) Delile

The main aim of this study was to estimate the effects of salinity variation on the Mediterranean seagrass Posidonia oceanica (L.) Delile and its attached epiphytes. Leaf growth and survival of this plant were tested in several short-term (15 days) mesocosms experiments under controlled conditions between February 2001 and November 2001. Plants collected from shallow meadows at Alicante (SE Spain), with an ambient salinity of 36.8-38.0 psu, were placed in tanks of 300 L with an additional overhead light and exposed to different salinity treatments (ranging from 25 to 57 psu) during 15 days. To estimate the mortality and growth recuperation, in some experiments shoots were returned to control salinity (38 psu). Leaf growth was measured in the laboratory where epiphytic fauna and flora were removed from leaves, with a razor blade, to determine their biomass.P. oceanica was negatively influenced by increased salinity. Shoots showed a significant decrease in growth and survival, whereas epiphyte biomass did not show a clear response because of their high variability. Maximum leaf growth occurred between 25 and 39 psu. In addition, plants suffered considerable mortality at salinities above 42 psu and below 29 psu, with 100% mortality at 50 psu. In salinities between 39 and 46 psu, surviving plants were able to regain their original growth rate when returned to normal seawater salinity (38 psu). These results suggest that P. oceanica is one of the most sensitive seagrasses to salinity increments it is more tolerant to salinity reductions (25.0-36.4 psu), perhaps due to the terrestrial origin of seagrasses.     

Motility of zooxanthellae isolated from the Red Sea soft coral Heteroxenia fuscescens (Cnidaria)

Unicellular dinoflagellate algae are among the best examples of organisms that exhibit biological clocks. This study examined the effect of light regime on rhythmicity of motility in the symbiotic dinoflagellate Symbiodinium sp., freshly isolated from the soft coral Heteroxenia fuscescens (Ehrenberg). Freshly isolated algal cells, placed under a 12-h L:12-h D cycle, exhibited motility with a diel rhythm. This motility occurred only during the period of illumination and lasted 8-9 h, with a peak at 2.5-4 h after lights on. Algal cells placed in an inverted light regime inverted their motility pattern. The response to the L/D regime was very precise, and even a 1-h shift backward or forward affected initiation of motility and time of its maximal peak. When placed in either constant light or dark, algal motility ceased until the L/D cycle was restored. These findings suggest that the rhythm is entrained by light cues and is not due to an endogenous circadian rhythm. Further, we provide evidence that the presence of juvenile hosts does not affect the algal motility pattern. These results offer the first evidence for the lack of impact by the host on rhythmicity of motility of free-living algal cells. The motility pattern found in freshly isolated algae may indicate the presence of light-induced diel rhythmicity in yet-to-be described free-living Symbiodinium.     

The role of epibotic bacteria from the surface of the soft coral Dendronephthya sp. in the inhibition of larval settlement

It has been suggested that bacteria associated with soft-bodied organisms are suggested to produce bioactive compounds against the attachment of invertebrate larvae and bacteria onto the surface of these organisms. Our recent study has demonstrated that epibiotic bacteria from the surface of the soft coral Dendronephthya sp. (Coelenterata: Octocoralia, Alcyonacea) inhibit the growth of bacteria commonly found in marine natural biofilms. In the present study, the effect of 11 epibiotic bacteria isolated from the surface of Dendronephthya sp. on larval settlement of the tubeworms Hydroides elegans was examined using laboratory bioassay. Among 11 bacterial isolates, 2 strains (18%) inhibited the larval settlement of H. elegans (Haswell), 4 strains (36%) were “inductive” to larvae and the remaining 5 strains (46%) were “non-inductive”. There was no correlation between the antifouling activities of bacterial isolates and their phylogenetic origin, i.e. closely related bacterial strains showed different effects on larval settlement of H. elegans. When all “inductive”, “non-inductive” and “inhibitive” bacterial isolates were mixed in a 1:1:1 ratio, the effect of the resultant multispecies film on larval settlement became “inhibitive”. Waterborne compounds of Vibrio sp. and an unidentified α-Proteobacterium, which suppressed the settlement of H. elegans and Bugula neritina (L.) larvae, were further investigated using size fractionation and bioassay-guided enzymatic analysis. It was found that antilarval settlement compounds from these bacteria were heat-stable polysaccharides with a molecular weight >100 kDa. The results indicate that the bacteria associated with the soft coral Dendronephthya sp. may contribute to the antifouling mechanisms of the soft-bodied organisms by producing compounds that are against bacterial growth and settlement of macrofoulers on the surface of their host.     

Phenomorphology and ecomorphological characters of Maytenus senegalensis L. Shrublands in the Iberian Peninsula: A comparison with other Mediterranean plant communities

This work sets out to establish ecomorphological and phenomorphological patterns in the constituent flora of an Iberian Maytenus senegalensis plant community in order to relate it with or separate it from other types of Mediterranean vegetation. This plant community contains tall shrubs, scrubs, palms and vines of a variety of families. Typical ecomorphological adaptations to the Mediterranean climate were observed in the community studied: an abundance of thorny species or species with tomentose leaves and photosynthetic stems, small leaves and a high presence of species with organs capable of post-fire regeneration. However, the community also showed hybrid characteristics between scrubland (predominantly malacophyllous leaves lasting 9 months on average and low biomass) and tall shrubland communities (predominance of phanerophytes up to 3 m in height, predominantly smooth bark, periodical reposition of leaves, mean lifespan of plants 25 years and mainly fleshy fruit). The community also presents an outstanding quantity of amphiphytes (29%). From a phenological point of view, the formation of buds in winter, flowering in spring, summer–autumn seed dispersal and leaf shedding in summer are typical of other Mediterranean communities, while fructification and growth lasting three seasons, as in the case of Maytenus shrubland, is not. The RVA phenophasic index (balance between reproductive and vegetative phenophases) with values of 0.4–2 shows an intermediate value between typical shrubland (0.3–1) and forest community (1–2) values. The APC index (period of phenophasic activity of the community) differentiates it from others, since it remains at 100% for seven months, three of which are winter months. Phenophasic patterns (overlapping, or not, of flower bud formation, flowering and growth) show how similar M. senegalensis shrubland is to forests because of the absence of phenophase pattern type A (characteristic in open-scrublands); in addition, we have detected a new pattern (type G), still regarded as exclusive to M. senegalensis. As regards the existence of coenomorphs (group of species with similar morpho-functional strategies), the studied community showed much greater diversity than other communities studied to date, with an Strategies Richness index (SRi) of 0.79 (11 groups covering the 14 species existing), pointing to the co-existence of a great variety of eco-phenomorphological strategies, each with a successful response to the same ecological conditions. Finally, some of the species in the community, e.g. Asparagus horridus, Chamaerops humilis and M. senegalensis, stand out because of their unique ecomorphological and phenological characteristics.     

Ontogenetic and environmental effects on otolith shape variability in three Mediterranean European eel (Anguilla anguilla, L.) local stocks

Otolith morphology is an efficient tool for the discrimination of fish stocks, populations and species when comparative genetic data are not available. Currently, the relationship between environmental factors and otolith shape is poorly characterized for the European eel (Anguilla anguilla), a highly migratory catadromous species constituting a single, randomly mating stock. The present study analyses the differences in otolith morphology between three Mediterranean eel local stocks from different environmental contexts (i.e. two brackish lagoons and one river). The relationship between otolith shape and otolith size was studied by means of Elliptic Fourier analysis and multivariate statistics. Otolith profile was digitally acquired and Cartesian coordinates were extracted. Partial Least Square (PLS) analysis pointed to continuous allometric growth in size and shape in otoliths from all three sites. In the three environments, shape variations occurred during growth as indicated by the presence of a significant and positive relationship between otolith size and the first PLS latent vector (i.e. which bears most of the information regarding otolith outline). Differences between smaller and larger sized otoliths were investigated using PLS Discriminant Analysis (PLSDA) and cluster analysis. Results indicate that otolith shape is highly uniform at smaller than at larger sizes. These shape differences apparently overlap the initial differentiation of the small otolith outlines acquired by eels during the growing phase as elvers in the marine environment. Data were discussed considering that the physical and chemical habitat variability in brackish lagoons and river could underlie a marked change in otolith shape during the animals' growth.     

Experimental analysis of the response and recovery of Zostera marina (L.) and Halodule wrightii (Ascher.) to repeated light-limitation stress

Seagrasses are considered important indicators of decline in water quality resulting in increased light attenuation that negatively influences their growth and survival. Chronic light-limitation interspersed with unpredictable acute attenuation events have had poorly understood effects on seagrass recovery dynamics. Zostera marina (eelgrass) and Halodule wrightii (shoalgrass) were subject to a matrix of light-deprivation events followed by recovery periods to mimic repeated acute shading events. Plant survival, morphology, biomass, chlorophyll content, and Fv/Fm were assessed over time to determine recovery. At the end of the experiment, all plants were harvested and species-specific treatment effects were determined. Significant differences due to treatments were noted in all parameters measured. In general, responses were similar for both life-stages and between species, suggesting similar physiological tolerance to repeated acute light-attenuation events. Only plants in treatments where light-deprivation was followed by a recovery interval of at least the same duration showed signs of long-term survival. Chlorophyll fluorescence (Fv/Fm) was an important metric for assessing recovery, but it failed to detect the onset of mortality in many plants. Other metrics of plant condition need to be assessed and coupled with chlorophyll fluorescence data to assess seagrass “health”. This is of particular importance in field studies, where the history of the plants is largely unknown.     

Effect of trimethyllead chloride on slowly activating (SV) channels in red beet (Beta vulgaris L.) taproots

The patch-clamp technique was used to examine the effect of trimethyllead chloride (Met₃PbCl) on SV channel activity in red beet (Beta vulgaris L.) taproot vacuoles. It was found that in the control bath the macroscopic currents showed the typical slow activation and a strong outward rectification of the steady-state currents. An addition of Met₃PbCl to the bath solution blocked, in a concentration-dependent manner, SV currents in red beet vacuoles. The time constant τ increased several times in the presence of 100 μM trimethyllead chloride at all voltages tested. When single channel properties were analyzed, only little channel activity could be recorded in the presence of 100 μM Met₃PbCl. Trimethyllead chloride decreased significantly (by about one order of magnitude) the open probability of single channels. The recordings of single channel activity obtained in the presence and absence of Met₃PbCl showed that organolead only slightly (by ca. 10%) decreased the unitary conductance of single channels. It was also found that Met₃PbCl diminished significantly the number of SV channel openings, whereas it did not change the opening times of the channels. Taken together, these results suggest that Met₃PbCl binding site is located outside the channel selectivity filter.     

The presence of exotic Hypnea flexicaulis (Rhodophyta) in the Mediterranean Sea as indicated by morphology, rbcL and cox1 analyses

Here we report the first finding of Hypnea flexicaulis Yamagishi and Masuda in the Mediterranean Sea (Lagoon of Venice, Italy), identified through molecular analyses using the plastid ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) and the mitochondrial protein-coding cytochrome c oxidase subunit I (cox1) genes. The phylogenetic reconstruction, based on rbcL + cox1 multiple alignment, showed that all specimens of H. flexicaulis from Venice, Korea, Philippines and Taiwan were included in a monophyletic group supported by a bootstrap value of 100%.It is highly probable that H. flexicaulis has been introduced from Indo-Pacific populations, in particular the Korean one, probably via ship traffic or shellfish transfers.The use of DNA barcoding combined with morphological observations was, in this case, a rapid way to identify this allochthonous species.     

Genetic variation of isolated and peripheral populations of Pinus sylvestris (L.) from glacial refugia

Analysis of variation at 10 polymorphic isoenzyme loci in 51 European populations of Pinus sylvestris both from its main continuous range and from isolated, marginal populations out of the continuous range, revealed differences in genetic structure. The highest genetic richness, measured as mean number of alleles per locus, was recorded for populations from the Iberian Peninsula, followed by those from Scotland and the Balkans. Marginal, isolated populations were characterized by much greater interpopulation variation than populations from the continuous range of the species. The highest mean observed heterozygosity was recorded for the Iberian populations. The highest gene flow was observed among populations in the continuous range of the species. The populations could be classified into five groups based on genetic similarities. Populations from the continuous range formed one group, those from Spain two groups, pines from Scotland one group and those from the Balkans one group. One population from Scotland was found to be most distinct from the other Scottish pines and was found to be grouped with the Balkan populations. The occurrence and frequencies of rare alleles in the populations showed a characteristic pattern, suggesting that populations from the Iberian Peninsula probably did not participate in the colonization of Europe by Pinus sylvestris after the last glaciation. In contrast, the migration of populations from the Balkan refugium into Central and Western Europe had a significant effect on the contemporary gene pools of populations of Pinus sylvestris in its continuous range.     

Changes in growth and reproduction of green sea urchins, Strongylocentrotus droebachiensis (Müller), during repopulation of the shallow subtidal zone after mass mortality

Following a disease outbreak that caused mass mortality of green sea urchins, Strongylocentrotus droebachiensis, along the Atlantic coast of Nova Scotia in September 1999, changes in growth and reproduction were monitored over 3.75 years as surviving individuals migrated from deep water to repopulate the shallow subtidal zone at a wave-exposed site. Urchins were sampled at 4 depth strata: at 24 m on a boulder field where the population was unaffected by the disease, at 12 and 16 m on a steeply sloping bedrock ramp, and at 8-10 m along the lower margin of a kelp bed (Laminaria digitata) where urchins formed a grazing front by January 2002. Urchins migrating shoreward from the deep-water refuge responded rapidly to increased algal productivity in the shallows through increased growth and reproduction. Measures of annual increments of skeletal elements (rotules) from urchins across the depth gradient indicated that the fastest growing individuals from the source population formed the grazing front. Urchins in the front reached a larger asymptotic size and produced larger gonads than urchins lower on the ramp. The annual cycle in gonad index showed a pronounced spring spawning period across all depths; a secondary fall spawning was evident at the front and 12 m. The presence of mature, fertilizable ova and short response time to spawning induction in both spring and fall supported the occurrence of two spawning periods.     

Ontogenetic- and condition-related effects of starvation on responsiveness in herring larvae (Clupea harengus L.) during repeated attacks by a model predator

The escape behaviour of fed and food-deprived herring larvae (20-30 mm SL) was studied during repeated attacks with a glass probe in order to investigate the causal mechanisms of starvation-induced changes in reaction to predators. Two experiments were set up: one where the fed and starved larvae were of the same age but of different sizes and one with groups of similar size but different ages. Biochemical and morphological condition measures described the condition of individual larvae. Starved larvae showed a lower responsiveness than fed larger larvae of the same age, and the responsiveness in this experiment decreased with decreasing nutritional status of the larvae. However, starved larvae that were of the same size but older than fed larvae showed a higher responsiveness, which could be explained by differences in development of sensory systems. A combination of condition and developmental factors thus explained the differences between starved and fed larvae. Both starved and fed larvae showed decreasing responsiveness over time with repeated attacks with no difference between starved and fed larvae in the relative change over time. The results emphasise the importance of taking into account individual age and/or development when the nutritional condition of wild-caught larvae is incorporated in survival models.