Isotopic niche provides an insight into the ecology of a symbiont during its geographic expansion.

The study of the recent colonization of a symbiont and its interaction with host communities in new locations is an opportunity to understand how they interact. The use of isotopic ratios in trophic ecology can provide measurements of a species' isotopic niche, as well as knowledge about how the isotopic niches between symbiont and host species overlap. Stable isotope measurements were used to assess the sources of carbon assimilated by the host species (the bivalves Mytilus galloprovincialis and Scrobicularia plana) and their associated symbiont pea crab Afropinnotheres monodi, which occurs within these bivalves' mantle cavities. The mixing model estimates suggest that all of them assimilate carbon from similar sources, particularly from pseudofaeces and particulate organic matter in this symbiotic system based on filter feeding. The symbiotic species occupy comparable trophic levels and its association seems to be commensal or parasitic depending on the duration of such association. The pea crab A. monodi reflects a sex-specific diet, where males are more generalist than the soft females because the latter's habitat is restricted to the host bivalve. The high isotopic overlap between soft females and M. galloprovincialis may reflect a good commensal relationship with the host.

Null alleles are ubiquitous at microsatellite loci in the Wedge Clam (Donax trunculus).

Recent studies have reported an unusually high frequency of nonamplifying alleles at microsatellite loci in bivalves. Null alleles have been associated with heterozygous deficits in many studies. While several studies have tested for its presence using different analytical tools, few have empirically tested for its consequences in estimating population structure and differentiation. We characterised 16 newly developed microsatellite loci and show that null alleles are ubiquitous in the wedge clam, Donax trunculus. We carried out several tests to demonstrate that the large heterozygous deficits observed in the newly characterised loci were most likely due to null alleles. We tested the robustness of microsatellite genotyping for population assignment by showing that well-recognised biogeographic regions of the south Atlantic and south Mediterranean coast of Spain harbour genetically different populations.

Implications for management and conservation of the population genetic structure of the wedge clam Donax trunculus across two biogeographic boundaries.

In a resource management perspective, the understanding of the relative influence of the physical factors on species connectivity remains a major challenge and is also of great ecological and conservation biology interest. Despite the overfishing threat on the wedge clam Donax trunculus in Europe, relatively little information is known about its population genetic structure and connectivity and their consequences on conservation policies. We employed 16 microsatellite loci to characterise the genetic diversity and population structure of D. trunculus. A total of 514 samples from seven different localities along the Atlantic-Mediterranean transition, from the Atlantic (Gulf of Cádiz) to the north-western Mediterranean were genotyped. The analysis of the population genetic structure displayed a clear distinction along the Atlantic-Mediterranean transition with different clusters in the Atlantic Ocean, the Alboran Sea and the northwestern Mediterranean. Consequently, we recommend that these three areas should be considered as different management units. We showed that all populations seem to be at high long-term risk of extinction with the exception of the protected Doñana National Park population which still seems to have evolutionary potential. Therefore, our results emphasized the necessity of protection of this economic resource and the validity of molecular tools to evaluate the population dynamics.

Influence of temperature on toxicity of single pharmaceuticals and mixtures, in the crustacean A. desmarestii.

Lethal and sublethal responses of the shrimp Atyaephyra desmarestii exposed to three pharmaceutical compounds, Diclofenac (DF), Ibuprofen (IB) and Carbamazepine (CBZ), individually and in mixtures, were evaluated under two temperature scenarios. LC50 (96h) values were obtained individually at 20° and 25°C. At 25°C, mortality in binary and ternary mixtures is higher than at 20°C. The toxicity of the mixtures was predicted on the basis of individual mortality data using two toxicity models: Concentration addition (CA) and Independent action (IA). Our results showed that neither CA nor IA unequivocally predicted the observed toxicity of binary and ternary mixtures. For sublethal toxicity, selected endpoints were: ingestion rate, osmoregulatory capacity and respiration rate. Regarding osmoregulatory capacity, no significant differences were found. The highest ingestion rates were recorded in organisms exposed at 25°C, irrespective of the compound, after 30 and 60min of exposure. At 20°C, there was a significant decrease in respiration rate (Dunnett́s test p<0.05) under conditions of severe anoxia (1mg O2L(-1)) in organisms exposed to 13.3µgL(-1) of DF. At 25°C a significantly lower respiration rate with respect to the control (Dunnett́s test p<0.05) was found in organisms exposed to 13.8µgL(-1) of CBZ under conditions of moderate hypoxia and well-oxygenated water (3 and 5mg O2L(-1), respectively). The respiratory independence of organisms exposed to the higher temperature (25°C) also decreased. This study shows that CBZ and DF individually, even at relatively low concentrations, may produce respiratory deficiencies in the freshwater shrimp, Atyaephyra desmarestii under certain temperature and water oxygenation conditions, thus reducing its ability to function.

A salt bath will keep you going? Euryhalinity tests and genetic structure of caridean shrimps from Iberian rivers.

We assessed the role of euryhalinity and life-history traits on the population genetic structure of the four main caridean shrimp species from the Iberian Peninsula (Atyaephyra desmarestii, Dugastella valentina, Palaemon varians and Palaemon zariquieyi) able to complete their life cycle in freshwater/oligohaline habitats. Seawater exposure experiments indicated that A. desmarestii, D. valentina and P. zariquieyi are more sensitive to high salinity waters than P. varians and confirm the relationship between osmolality regulation and spatial distribution of species. The limited or no survival in seawater could explain the restricted distributions observed in D. valentina and P. zariquieyi, whereas the current A. desmarestii distribution could be due to either past river dynamics and/or human-mediated water transfers. Conversely, the high tolerance of P. varians to a large salinity range (euryhalinity) could explain its capacity to colonize geographically distant estuaries. In agreement with osmoregulation results, the phylogeography patterns of the cytochrome oxidase 1 (Cox 1) gene fragment revealed significant genetic differentiation among river systems whatever the species considered. Atyidae species presented higher nucleotide diversity levels than Palaemonidae species, while isolation-by-distance patterns were only found for the latter. Our results have important implications for the management and conservation of freshwater species, since the inter-catchment connectivity may affect the speciation processes.

Freshwater scarcity effects on the aquatic macrofauna of a European Mediterranean-climate estuary.

In the Mediterranean-climate zone, recurrent drought events and increasing water demand generally lead to a decrease in freshwater input to estuaries. This water scarcity may alter the proper function of estuaries as nursery areas for marine species and as permanent habitat for estuarine species. A 12-year data set of the aquatic macrofauna (fish, decapod and mysid crustaceans) in a Mediterranean estuary (Guadalquivir estuary, South Spain) was analysed to test if water scarcity favours the nursery function of regional estuaries to the detriment of permanent estuarine inhabitants. Target species typically displayed a salinity-related distribution and estuarine salinisation in dry years resulted in a general upstream community displacement. However, annual densities of marine species were neither consistently higher in dry years nor estuarine species during wet years. Exceptions included the estuarine mysid Neomysis integer and the marine shrimp Crangon crangon, which were more abundant in wet and dry years, respectively. High and persistent turbidity, a collateral effect of water scarcity, altered both the structural (salinity-related pattern) and functional (key prey species and predator density) community characteristics, chiefly after the second drought period of the analysis. The observed high inter-year environmental variability, as well as species-specific effects of water scarcity, suggests that exhaustive and long-term sampling programmes will be required for rigorously monitoring the estuarine communities of the Mediterranean-climate region.

Is Atyaephyra desmarestii a useful candidate for lethal and sub-lethal toxicity tests on pharmaceutical compounds?

Single and mixture toxicity tests on three pharmaceutical compounds, Diclofenac (DF), Ibuprofen (IB) and Carbamazepine (CBZ), were carried out with the freshwater shrimp Atyaephyra desmarestii. Lethal and sublethal responses were analyzed for single compounds. Lethal concentrations (LC50) obtained for each individual compound, after 96 h of exposure, were 6.3 mg L(-1) for DF, 13.3 mg L(-1) for IB and 94.3 mg L(-1) for CBZ. The selected sublethal endpoints of food ingestion, osmoregulatory capacity and respiration rates were not affected by the exposures to environmentally-relevant concentrations. Based on mortality data obtained, the predictive no effect concentration (PNEC) was calculated for each of the compounds, and compared with predicted environmental concentrations (PEC) reported in surface waters. The environmental risk of each compound was estimated as the ratio between PEC/PNEC, and indicated that IB could represent a medium risk in freshwater environments. Additionally, binary and ternary mixture toxicity assays of the selected compounds were carried out. The data obtained was applied to two predictive toxicity models: Concentration Addition (CA) and Independent Action (IA). Finally, risk assessment was estimated using risk quotients (RQ) for the compound mixtures based on EC50 and LC50 values.

Keratocyte density 3 months, 15 months, and 3 years after corneal surface ablation with mitomycin C.

PURPOSE: To study the effects of surface ablation with mitomycin C (MMC) on keratocyte population. DESIGN: Prospective, nonrandomized, interventional, comparative case series. METHODS: Thirty two eyes treated with surface ablation with 0.02% MMC were compared with nontreated eyes at Vissum Santa Hortensia, Madrid, Spain. Keratocyte density was measured with the Heidelberg Retina Tomograph II (Rostock Cornea Module) 3, 15, and 36 to 42 months after the surgery in the anterior, mid, and posterior stroma, and compared with control eyes. RESULTS: Three months postoperatively, we found a lower stromal bed density compared to controls (16 993 ± 8001 vs 29 660 ± 5904 cells/mm(3), P = .0001), while there was a significantly higher cell density in the mid (30 783 ± 9300 vs 18 505 ± 1996 cells/mm(3), P = .0001) and deep stroma (30 268 ± 8321 vs 18 438 ± 2139 cells/mm(3), P = .0001). Three years after the surgery, the cellularity in the stromal bed had not significantly changed from the 3-month follow-up, but the density in the mid (18 889 ± 3474 cells/mm(3)) and posterior stroma (18 992 ± 3402 cells/mm(3)) had decreased to show no difference from controls. The mean cell density between the anterior, mid, and posterior stroma was not significantly different from controls 15 months and 3 years after the surgery. CONCLUSION: Our study suggests that there is a reorganization of the stromal cell population soon after surface ablation with MMC, with a decrease in the stromal bed compensated initially with an increase in the mid and posterior stroma. Corneal cellularity tends to normalize over time, and 3 years postoperatively the mean cell density throughout the cornea seems to maintain normal values.

Keratocyte density after laser-assisted subepithelial keratectomy with mitomycin C.

PURPOSE: To study the effects of laser-assisted subepithelial keratectomy (LASEK) with mitomycin C (MMC) on the keratocyte population. DESIGN: Prospective, nonrandomized, interventional, comparative case series. METHODS: Fifty-six eyes treated at Vissum Santa Hortensia, Madrid, Spain, were included in the study. We compared 28 eyes treated with LASEK with intraoperative 0.02% MMC versus 28 non-treated eyes. Keratocyte density was measured 3 months after the surgery in the anterior, mid, and posterior stroma and was compared with the corresponding layers in the control eyes. The anterior layer in the LASEK group was compared with 2 layers in the control group: the most anterior stromal layer and the 80 µm-deep layer, because that was the mean ablation depth performed in eyes that underwent LASEK. RESULTS: We found a statistically significantly lower keratocyte population in the most anterior stromal layer after LASEK with MMC compared with both the most anterior stromal layer and the 80 µm-deep layer in controls. On the contrary, the treated group showed a significantly higher keratocyte density in both the mid stroma and the deep stroma. The comparison between the average densities through the entire cornea showed a significantly higher keratocyte population in the LASEK with MMC group. CONCLUSIONS: LASEK with MMC seems to cause a decrease in the anterior stromal cells 3 months after the surgery compared with nonoperated corneas. There seems to be a compensating proliferation of keratocytes in the deeper corneal layers, suggesting that the ability of keratocytes to repopulate the cornea is maintained after the surgical procedure.

Comparison between femtosecond laser-assisted sub-Bowman keratomileusis vs laser subepithelial keratectomy to correct myopia.

PURPOSE: To compare femtosecond laser sub-Bowman keratomileusis (FSBK) vs laser subepithelial keratectomy (LASEK) to correct myopia. DESIGN: Retrospective, nonrandomized, interventional, comparative case series. METHODS: Two thousand one hundred and eight eyes were included in the study. We compared 1,072 eyes treated with FSBK vs 1,036 eyes treated with LASEK with or without mitomycin C (MMC). Visual and refractive results were evaluated 1 day, 1 week, and 1 and 3 months postoperatively. RESULTS: Preoperative mean sphere and best spectacle-corrected visual acuity (BSCVA) were -3.93 diopters (D) vs -3.87 D (P = .5) and 1.12 vs 1.12 (P = .8) in FSBK and LASEK, respectively. Uncorrected visual acuity (UCVA) was 0.92 vs 0.62, 0.98 vs 0.78, 0.96 vs 0.91, and 1.06 vs 1.03 in FSBK and LASEK, respectively, at 1 day, 1 week, and 1 and 3 months after surgery (P < .01 for all comparisons). Three months postoperatively, BSCVA was 1.13 and 1.10, respectively (P = .001). At that moment, 20 eyes (1.93%) in the LASEK group vs 9 eyes (0.84%) in the FSBK group had lost 2 or more lines of BSCVA. Ten eyes (0.96%) in the LASEK group gained 2 or more lines of BSCVA, whereas 3 eyes (0.28%) in the FSBK group gained 2 lines. Six months postoperatively, only 2 LASEK eyes (0.19%) showed loss of 2 or more lines of BSCVA, compared to 3 FSBK-treated eyes (0.28%). CONCLUSION: Both FSBK and LASEK are safe and effective procedures to correct myopia. Slightly better visual and refractive results were observed in FSBK-treated eyes in a 3-month follow-up.

Physiological and behavioral responses of the mud snails Hydrobia glyca and Hydrobia ulvae to extreme water temperatures and salinities: implications for their spatial distribution within a system of temperate lagoons.

Physiological responses (oxygen consumption) and behavioral responses (feeding and activity) of the mud snails Hydrobia ulvae and Hydrobia glyca at different salinities (20 per thousand-80 per thousand) and temperatures (20 degrees and 30 degrees C) were studied. After 24 h under experimental conditions, both Hydrobia species already showed maximal activities (>90%) for a wide salinity range (30 per thousand-70 per thousand), with significant differences in activity between species only outside the usual salinity range of the studied lagoon. In contrast, egestion rates of H. glyca were significantly higher at the lowest salinities tested (30 per thousand and 40 per thousand) irrespective of water temperature, whereas egestion rates of H. ulvae were always significantly higher (57% on average) at 20 degrees C than at 30 degrees C and at the usual salinities found in the field (40 per thousand and 50 per thousand). Both species showed an oxyregulatory response to dissolved oxygen concentrations ranging from saturation to 1.5 mg O(2) L(-1), although specific oxygen consumption rates were significantly higher at 30 degrees C than at 20 degrees C (Q(10)=1.47+/-0.08 for H. ulvae and Q(10)=12.1+/-0.06 for H. glyca) and at the lowest salinities (30 per thousand-50 per thousand for H. ulvae and 30 per thousand-40 per thousand for H. glyca). On average, specific rates were higher for the smaller-sized H. glyca (1.64+/-0.03 microg O(2) mg(-1) ash-free dry weight [AFDW]) than for H. ulvae (1.35+/-0.03 microg O(2) mg(-1) AFDW). Despite the overlapping of their tolerances to high temperatures and salinities, the observed interspecies differences could play a certain role in the distribution of H. ulvae and H. glyca in the studied habitat. In particular, the decreasing feeding activity but increasing respiration of H. ulvae at 30 degrees C for salinities that usually occur in the studied lagoon could represent disadvantages to H. glyca during the warm period.

Oxygen consumption and osmoregulatory capacity in Neomysis integer reduce competition for resources among mysid shrimp in a temperate estuary.

Results of field surveys and laboratory measurements of oxygen consumption and body fluid osmolality at different salinities in the mysids Neomysis integer, Mesopodopsis slabberi, and Rhopalophthalmus mediterraneus from the Guadalquivir estuary (southwest Spain) were used to test the hypothesis that osmotic stress (oxygen consumption vs. isosmotic points) was lowest at salinities that field distributions suggest are optimal. The three species showed overlapping spatial distributions within the estuary but clear segregation along the salinity gradient: N. integer, M. slabberi, and R. mediterraneus displayed maximal densities at lower, intermediate, and higher salinities, respectively. Adults of N. integer were extremely efficient hyperregulators (isosmotic point 30 per thousand) over the full salinity range tested (3 per thousand-32 per thousand), and their oxygen consumption rates were independent of salinity; adults of M. slabberi were strong hyper- and hyporegulators at salinities between 7 per thousand and 29 per thousand (isosmotic point, 21 per thousand) and showed higher oxygen consumptions at the lowest salinity (6 per thousand); adults of R. mediterraneus hyperregulated at salinities between 19 per thousand and seawater (isosmotic point, 36 per thousand), with the lowest oxygen consumption at salinity around their isosmotic point (35 per thousand). Thus, the osmoregulation capabilities of M. slabberi and R. mediterraneus seem to determine the salinity ranges in which most of their adults live, but this is not so for adults of N. integer. Moreover, maximal field densities of M. slabberi (males and females) and R. mediterraneus (males) occur at the same salinities as the lowest oxygen consumption. In contrast, field distribution of N. integer was clearly biased toward the lower end of the salinity ranges within which it osmoregulated. We hypothesize that the greater euryhalinity of N. integer makes it possible for this species to avoid competition with R. mediterraneus by inhabiting the more stressful oligohaline zone.