Characterizing niche differentiation among marine consumers with amino acid δ13C fingerprinting

Marine food webs are highly compartmentalized, and characterizing the trophic niches among consumers is important for predicting how impact from human activities affects the structuring and functioning of marine food webs. Biomarkers such as bulk stable isotopes have proven to be powerful tools to elucidate trophic niches, but they may lack in resolution, particularly when spatiotemporal variability in a system is high. To close this gap, we investigated whether carbon isotope (δ¹³C) patterns of essential amino acids (EAAs), also termed δ¹³C_AA fingerprints, can characterize niche differentiation in a highly dynamic marine system. Specifically, we tested the ability of δ¹³C_AA fingerprints to differentiate trophic niches among six functional groups and ten individual species in the Baltic Sea. We also tested whether fingerprints of the common zooplanktivorous fishes, herring and sprat, differ among four Baltic Sea regions with different biochemical conditions and phytoplankton assemblages. Additionally, we investigated how these results compared to bulk C and N isotope data for the same sample set. We found significantly different δ¹³C_AA fingerprints among all six functional groups. Species differentiation was in comparison less distinct, due to partial convergence of the species' fingerprints within functional groups. Herring and sprat displayed region‐specific δ¹³C_AA fingerprints indicating that this approach could be used as a migratory marker. Niche metrics analyses showed that bulk isotope data had a lower power to differentiate between trophic niches than δ¹³C_AA fingerprinting. We conclude that δ¹³C_AA fingerprinting has a strong potential to advance our understanding of ecological niches, and trophic linkages from producers to higher trophic levels in dynamic marine systems. Given how management practices of marine resources and habitats are reshaping the structure and function of marine food webs, implementing new and powerful tracer methods are urgently needed to improve the knowledge base for policy makers.     

Assessing the genetic diversity in Argopecten nucleus (Bivalvia: Pectinidae), a functional hermaphrodite species with extremely low population density and self‐fertilization: Effect of null alleles

Argopecten nucleus is a functional hermaphroditic pectinid species that exhibits self‐fertilization, whose natural populations have usually very low densities. In the present study, the genetic diversity of a wild population from Neguanje Bay, Santa Marta (Colombia), was estimated using microsatellite markers, and the effect of the presence of null alleles on this estimation was assessed. A total of 8 microsatellite markers were developed, the first described for this species, and their amplification conditions were standardized. They were used to determine the genotype of 48 wild individuals from Naguanje Bay, and 1,010 individuals derived from the offspring of 38 directed crosses. For each locus, the frequencies of the identified alleles, including null alleles, were estimated using the statistical package Micro‐Checker, and the parental genotypes were confirmed using segregation analysis. Three to 8 alleles per locus with frequencies from 0.001 to 0.632 were detected. The frequencies of null alleles ranged from 0.10 to 0.45, with Ho from 0.0 to 0.79, and He from 0.53 to 0.80. All loci were in H‐W disequilibrium. The null allele frequencies values were high, with lower estimations using segregation analysis than estimated using Micro‐Checker. The present results show high levels of population genetic diversity and indicate that null alleles were not the only cause of deviation from H‐W equilibrium in all loci, suggesting that the wild population under study presents signs of inbreeding and Wahlund effect.     

Spatial and temporal variability in ringed seal (Pusa hispida) stable isotopes in the Beaufort Sea

Arctic ecosystem dynamics are shifting in response to warming temperatures and sea ice loss. Such ecosystems may be monitored by examining the diet of upper trophic level species, which varies with prey availability. To assess interannual variation in the Beaufort Sea ecosystem, we examined spatial and temporal trends in ringed seal (Pusa hispida) δ¹³C and δ¹⁵N in claw growth layers grown from 1964 to 2011. Stable isotopes were correlated with climate indices, environmental conditions, seal population productivity, and geographic location. Sex and age did not influence stable isotopes. Enriched ¹³C was linked to cyclonic circulation regimes, seal productivity, and westward sampling locations. Higher δ¹⁵N was linked to lower sea surface temperatures, a higher percentage of pups in the subsistence harvest, and sample locations that were eastward and further from shore. From the 1960s to 2000s, ringed seal niche width expanded, suggesting a diversification of diet due to expansion of prey and/or seal space use. Overall, trends in ringed seal stable isotopes indicate changes within the Beaufort Sea ecosystem affected by water temperatures and circulation regimes. We suggest that continued monitoring of upper trophic level species will yield insights into changing ecosystem structure with climate change.     

Acidocalcisomes

Acidocalcisomes are acidic organelles containing calcium and a high concentration of phosphorus in the form of pyrophosphate (PP_i) and polyphosphate (poly P). Organelles with these characteristics have been found from bacteria to human cells implying an early appearance and persistence over evolutionary time or their appearance by convergent evolution. Acidification of the organelles is driven by the presence of vacuolar proton pumps, one of which, the vacuolar proton pyrophosphatase, is absent in animals, where it is substituted by a vacuolar proton ATPase. A number of other pumps, antiporters, and channels have been described in acidocalcisomes of different species and are responsible for their internal content. Enzymes involved in the synthesis and degradation of PP_i and poly P are present within the organelle. Acidocalcisomes function as storage sites for cations and phosphorus, and participate in PP_i and poly P metabolism, calcium homeostasis, maintenance of intracellular pH, and osmoregulation. Experiments in which the acidocalcisome Ca²⁺-ATPase of different parasites were downregulated or eliminated, or acidocalcisome Ca²⁺ was depleted revealed the importance of this store in Ca²⁺ signaling needed for host invasion and virulence. Acidocalcisomes interact with other organelles in a number of organisms suggesting their association with the endosomal/lysosomal pathway, and are considered part of the lysosome-related group of organelles.     

Overview of sperm whale Physeter macrocephalus mortality events in the Adriatic Sea, 1555–2009

• 1 In the Mediterranean Sea, the sperm whale Physeter macrocephalus is one of eight regular cetacean species. Poor knowledge of its ecology and status, together with suspected decline in numbers, make studies of historical and present occurrence especially relevant. Long‐term time series of stranding events are the most reliable data to provide a scientific framework for testing hypotheses that seek to explain the mechanisms responsible for cetacean strandings.
• 2 We present a comprehensive overview of cases of sperm whale mortality and human response to such events encompassing five centuries (1555–2009) within a portion of the Mediterranean Sea that offers a wealth of historical information – the Adriatic Sea.
• 3 A total of 36 mortality events were validated, involving 68 animals. Two findings of skeletal materials are also reported. The geographic distribution of strandings within the basin clearly was uneven: 44% of records (n = 16) were clustered along a 280km portion of the western Adriatic coast. A relatively high number of mortality events occurred along gently sloping sandy beaches away from suitable sperm whale habitat.
• 4 Until the first half of the 20th century, live‐stranded animals were routinely killed: all but one cases with known human response elicited killing attempts. Starting from the 1980s, killing was replaced by efforts to rescue the animals.
• 5 Mass strandings of sperm whales have occurred since historical times in the Adriatic Sea. Mortality events involving multiple individuals accounted for at least 17% of the total sample (6 of 36 mortality events). At least 29% of live strandings (6 of 21) involved more than one individual.
• 6 This study contributes a long‐term dataset based on careful validation of historical information, suitable for hypothesis testing aimed at investigating spatial and temporal correlates of sperm whale strandings – particularly live strandings – as a clue to their causes.

     

Factors leading to cannibalism in Lytechinus variegatus (Echinodermata: Echinoidia) held in intensive culture

Ecologically, sea urchins are an important species in marine habitats around the world. Economically, sea urchins are prized for their gonads (uni). With fisheries declining worldwide, intensive sea urchin culture has been proposed. For urchins in commercial culture, any factor that affects survivorship negatively should be addressed to maximize cost benefit. One potential obstacle to optimizing culture of sea urchins is cannibalism. Approximately 2000 adult and juvenile Lytechinus variegatus (1 g-45 g) were collected from Port Saint Joseph Peninsula State Park, FL between June and September 2009. Urchins were held in recirculating tanks at different sizes, densities, and feeding regimes for 4 weeks. Starvation and high density contributed to the highest level of cannibalism among small (12-21 g) urchins (percent cannibalism = 18.8%), whereas fed, high density conditions contributed to the highest level of cannibalism among large (32-37 g) urchins (percent cannibalism = 18.4%). These results suggest that (1) small urchins cannibalize at higher rates than large urchins, and (2) increased density is an important contributing factor leading to cannibalism. We quantified stress, defined as a decrease in production as a result of environmental conditions, by evaluating weight gain within each treatment and suggest that weight loss or minimal weight gain is an indicator of stress. We hypothesize increased stress caused by competitive interference can lead to increased cannibalism and decreased growth rates, even when food is not limiting. Ecologically, there are no reports of cannibalism of urchins in wild populations. Consequently, the role of cannibalism in regulating sea urchin community structure is not known. However, factors affecting cannibalism of L. variegatus in the laboratory may provide insight into the conditions that could result in cannibalism in wild populations. From an aquaculture perspective, it is important to determine those factors that contribute to the incidence of cannibalism in sea urchins so that the appropriate culture conditions can be maintained to reduce the incidence of cannibalism.     

Influence of short-term nutritional variations on digestive enzyme and fatty acid patterns of the calanoid copepod Temora longicornis

Temora longicornis, a dominant calanoid copepod species in the North Sea, is characterised by low lipid reserves and high biomass turnover rates. To survive and reproduce successfully, this species needs continuous food supply and thus requires a highly flexible digestive system to exploit various food sources. Information on the capacity of digestive enzymes is scarce and therefore the aim of our study was to investigate the enzymatic capability to respond to quickly changing nutritional conditions. We conducted two feeding experiments with female T. longicornis from the southern North Sea off Helgoland. In the first experiment in 2005, we tested how digestive enzyme activities and enzyme patterns as revealed by substrate SDS-PAGE (sodium dodecylsulfate polyacrylamide gel electrophoresis) responded to changes in food composition. Females were incubated for three days fed ad libitum with either the heterotrophic dinoflagellate Oxyrrhis marina or the diatom Thalassiosira weissflogii. At the beginning and at the end of the experiment, copepods were deep-frozen for analyses. The lipolytic enzyme activity did not change over the course of the experiment but the enzyme patterns did, indicating a distinct diet-induced response. In a second experiment in 2008, we therefore focused on the enzyme patterns, testing how fast changes occur and whether feeding on the same algal species leads to similar patterns. In this experiment, we kept the females for 4 days at surplus food while changing the algal food species daily. At day 1, copepods were offered O. marina. On day 2, females received the cryptophycean Rhodomonas baltica followed by T. weissflogii on day 3. On day 4 copepods were again fed with O. marina. Each day, copepods were frozen for analysis by means of substrate SDS-PAGE. This showed that within 24 h new digestive enzymes appeared on the electrophoresis gels while others disappeared with the introduction of a new food species, and that the patterns were similar on day 1 and 4, when females were fed with O. marina. In addition, we monitored the fatty acid compositions of the copepods, and this indicated that specific algal fatty acids were quickly incorporated. With such short time lags between substrate availability and enzyme response, T. longicornis can successfully exploit short-term food sources and is thus well adapted to changes in food availability, as they often occur in its natural environment due seasonal variations in phyto- and microzooplankton distribution.     

Burrow ventilation and associated porewater irrigation by the polychaete Marenzelleria viridis

Burrow ventilation of benthic infauna generates water currents that irrigate the interstices of the sediments surrounding the burrow walls. Such activities have associated effects on biogeochemical processes affecting ultimately important ecosystem processes. In this study, the ventilation and irrigation behavior of Marenzelleria viridis, an invasive polychaete species in Europe, was analyzed using different approaches. M. viridis showed to perform two types of ventilation: (1) muscular pumping of water out of the burrow and (2) cilia pumping of water into the burrow. Flowmeter measurements presented muscular pumping in time averaged rates of 0.15 ml min⁻¹. Oxygen needle electrodes positioned above the burrow openings revealed that muscular undulation of the worm body pumps anoxic water out of the burrow. On the other hand, microscope observations of the animal showed that ventilation of oxygen-rich water in the burrow occurs by ciliary action. The volume of water irrigated by M. viridis appears to vary linearly within the first 24 h incubation, with rates ranging from 0.003 to 0.01 ml min⁻¹. From those rates we could estimate that the time averaged rate of cilia ventilation should be about 0.16 ml min⁻¹. Since the cilia pumping into the burrow occurs in periods of 24 ± 12 min and at 50-70% of the measured time, considerable amounts of water from deeper sediments may percolate upwards to the sediment surface. This water is rich in reduced compounds and nutrients and may have important associated ecological implications in the ecosystem (e.g. affecting redox conditions, organic matter degradation, benthic recruitment and primary production).     

Phylogeography of the Atlanto-Mediterranean sea cucumber Holothuria (Holothuria) mammata: the combined effects of historical processes and current oceanographical pattern

We assessed the genetic structure of populations of the widely distributed sea cucumber Holothuria (Holothuria) mammata Grube, 1840, and investigated the effects of marine barriers to gene flow and historical processes. Several potential genetic breaks were considered, which would separate the Atlantic and Mediterranean basins, the isolated Macaronesian Islands from the other locations analysed, and the Western Mediterranean and Aegean Sea (Eastern Mediterranean). We analysed mitochondrial 16S and COI gene sequences from 177 individuals from four Atlantic locations and four Mediterranean locations. Haplotype diversity was high (H=0.9307 for 16S and 0.9203 for COI), and the haplotypes were closely related (π=0.0058 for 16S and 0.0071 for COI). The lowest genetic diversities were found in the Aegean Sea population. Our results showed that the COI gene was more variable and more useful for the detection of population structure than the 16S gene. The distribution of mtDNA haplotypes, the pairwise F(ST) values and the results of exact tests and amova revealed: (i) a significant genetic break between the population in the Aegean Sea and those in the other locations, as supported by both mitochondrial genes, and (ii) weak differentiation of the Canary and Azores Islands from the other populations; however, the populations from the Macaronesian Islands, Algarve and West Mediterranean could be considered to be a panmictic metapopulation. Isolation by distance was not identified in H. (H.) mammata. Historical events behind the observed findings, together with the current oceanographic patterns, were proposed and discussed as the main factors that determine the population structure and genetic signature of H. (H.) mammata.