Search

The protandric shrimp Hippolyte inermis is the only known marine invertebrate whose sex determination is strongly influenced by the composition of its food. In H. inermis, a sex reversal is triggered by the ingestion of diatoms of the genus Cocconeis associated with leaves of the seagrass Posidonia oceanica. These diatoms contain compounds that promote programmed cell death (PCD) in H. inermis and also in human cancer cells. Transcriptomic analyses suggested that ferroptosis is the primary trigger of the shrimp’s sex reversal, leading to the rapid destruction of the androgen gland (AG) followed by a chain of apoptotic events transforming the testes into ovaries. Here, we propose a molecular approach to detect the effects of compounds stimulating the PCD. An RNA extraction method, suitable for young shrimp post-larvae (five days after metamorphosis; PL5 stage), was established. In addition, six genes involved in apoptosis, four involved in ferroptosis, and seven involved in the AG switch were mined from the transcriptome, and their expression levels were followed using real-time qPCR in PL5 fed on Cocconeis spp., compared to PL5 fed on a basic control feed. Our molecular approach, which detected early signals of sex reversal, represents a powerful instrument for investigating physiological progression and patterns of PCD in marine invertebrates. It exemplifies the physiological changes that may start a few days after the settlement of post-larvae and determine the life destiny of an individual.

A considerable amount of coastal contamination is caused by wastes deriving from household and the degradation and the metabolism of plants and animals, even if our attention is commonly focused on industrial pollutants and contaminants. Waste pollutants are mainly represented by highly diluted soluble compounds and particles deriving from dead organisms. This complex combination, consisting of suspended particles and dissolved nutrients, has a significant impact on coastal planktonic and benthic organisms, also playing an active role in the global cycles of carbon. In addition, production practices are nowadays shifting towards recirculated aquaculture systems (RAS) and the genic responses of target organisms to the pollution deriving from animal metabolism are still scarcely addressed by scientific investigations. The reservoir of organic matter dissolved in the seawater is by far the least understood if compared to that on land, cause only a few compounds have been identified and their impacts on animals and plants are poorly understood. The tendency of these compounds to concentrate at interfaces facilitates the absorption of dissolved organic compound (DOC) onto suspended particles. Some DOC components are chemically combined with dissolved metals and form complexes, affecting the chemical properties of the seawater and the life of the coastal biota. In this research, we compared the reproductive performances of the common sea urchin Paracentrotus lividus cultured in open-cycle tanks to those cultured in a recirculating aquaculture system (RAS), where pollution progressively increased during the experiment due to animal escretions. Sea urchins were cultured for 7 months under these two conditions and their gametes were collected. Embryos resulting by in vitro fertilization were analyzed by Real Time qPCR to identify possible effects of pollution-induced stress. The fertility of sea urchins was evaluated, as well as the gonadosomatic indices and the histological features of gonads. Our results indicate that pollution due to excess of nutrients, event at sub-lethal concentrations, may hardly impact the reproductive potential of this key species and that chronic effects of stress are revealed by the analyses of survival rates and gene expression.

Communication among marine organisms are generally based on production, transmission, and interpretation of chemical cues. Volatile organic compounds (VOCs) can act as infochemicals, and ocean acidification can alter their production in the source organisms as well as the interpretation of the information they drive to target organisms. Two diatoms (Cocconeis scutellum var. parva and Diploneis sp.) and a macroalga (Ulva prolifera), all common epiphytes of Posidonia oceanica leaves, were isolated and cultured at two pH conditions (8.2 and 7.7). Their biomass was collected, and the VOCs produced upon wounding were extracted and analyzed using gas chromatography. Chemotactic reactions of invertebrates triggered by VOCs were tested using a static choice experimental arena and a flow-through flume system. Odor choice experiments were performed on several invertebrates associated with P. oceanica meadows to investigate the modification of behavioral responses due to the growth of algae in acidified environments. Complex patterns of behavioral responses were recorded after exposure to algal VOCs. This study demonstrated that a) ocean acidification alters the bouquet of VOCs released by diatoms and macroalgae and b) these compounds act as infochemicals and trigger peculiar behavioral responses in benthic invertebrates. In addition, behavioral responses are species-specific, dose-dependent, and are modified by environmental constraints. In fact, the static diffusion in choice arenas produced different responses as compared to flow-through flume systems. In conclusion, we demonstrate that in future marine environments higher CO2 concentrations (leading to a pH 7.7 by the end of this century) will modify the production of VOCs by micro- and macroalgae as well as the recognition of these infochemicals by marine invertebrates.

In recent decades, various species of Mediterranean sea urchins, including Paracentrotus lividus, have been subject to widespread seasonal episodes of mass mortality whose causative agents are still unclear. In particular, P. lividus is subject to late winter events of mortality, due to a disease manifested by a massive loss of spines and the presence of greenish amorphous material on the tests (i.e., the sea urchin skeleton consisting of spongeous calcite). Documented mortality events show a seasonal epidemic diffusion and might produce economic losses also in aquaculture facilities, besides the environmental constraints to its diffusion. We collected individuals showing conspicuous lesions on the body surface and reared them in recirculated aquaria. Samples of external mucous were collected along with coelomic liquids and cultured to isolate bacterial and fungal strains, further submitted to molecular identification through the amplification of prokaryotic 16S rDNA. In addition, pools of infected sea urchins were reared in recirculated tanks after short baths in a formulated therapeutic compound and their survival rates were compared to non-treated individuals for variable periods. Here, we aimed at a redescription of the etiopathogenetic nature of the parasites and tested the efficacy of a possible treatment, to be proposed for aquaculture purposes.

Paracentrotus lividus gonads, often referred to as “roe” or “uni” for gastronomical purposes, are among the most appreciated seafood delicacies in the Mediterranean area and worldwide. However, the increasing demand for human consumption has caused a growing pressure on its wild stocks, prompting the need to develop efficient aquaculture systems for its production. The set-up of effective feeds for various life stages and optimal procedures for breeding post-larvae and adult sea urchins still need to be improved. Here, for the first time, we aim at developing artificial feeds for the post-larvae of P. lividus because a critical step to improve our productive skills is post-larval growth. We tested various natural and prepared components to speed up the growth and enhance the survival rates of post-larvae, while taking into account the abiotic and biotic factors influencing the culture conditions in three replicate sets of tanks, characterized by different volumes. We tested formulated feeds and compared them with the effect of fresh foods in the frame of canonical culture practices. Our results indicated the efficiency of a feed composed of spirulina and Ulva rigida. Statistical analyses demonstrated the positive impact of this diet on the growth of post-larvae, behavior and survival rates. In addition, we demonstrated the efficacy of prepared feeds in the sea urchin aquaculture system, because they facilitated manipulation and control of the culture procedures for the satisfactory growth of P. lividus post-larvae.

Monitoring of aquatic ecosystems has been historically accomplished by intensive campaigns of direct measurements (by probes and other boat instruments) and indirect extensive methods such as aero-photogrammetry and satellite detection. These measurements characterized the research in the last century, with significant but limited improvements within those technological boundaries. The newest advances in the field of smart devices and increased networking capabilities provided by emerging tools, such as the Internet of Things (IoT), offer increasing opportunities to provide accurate and precise measurements over larger areas. These perspectives also correspond to an increasing need to promptly respond to frequent catastrophic impacts produced by drilling stations and intense transportation activities of dangerous materials over ocean routes. The shape of coastal ecosystems continuously varies due to increasing anthropic activities and climatic changes, aside from touristic activities, industrial impacts, and conservation practices. Smart buoy networks (SBNs), autonomous underwater vehicles (AUVs), and multi-sensor microsystems (MSMs) such as smart cable water (SCW) are able to learn specific patterns of ecological conditions, along with electronic “noses”, permitting them to set innovative low-cost monitoring stations reacting in real time to the signals of marine environments by autonomously adapting their monitoring programs and eventually sending alarm messages to prompt human intervention. These opportunities, according to multimodal scenarios, are dramatically changing both the coastal monitoring operations and the investigations over large oceanic areas by yielding huge amounts of information and partially computing them in order to provide intelligent responses. However, the major effects of these tools on the management of marine environments are still to be realized, and they are likely to become evident in the next decade. In this review, we examined from an ecological perspective the most striking innovations applied by various research groups around the world and analyzed their advantages and limits to depict scenarios of monitoring activities made possible for the next decade.

Cyanobacteria may live in the water column and in the benthos of aquatic environments, or be symbionts of other organisms, as in the case of Phormidium-like cyanobacteria, known to influence the ecology of freshwater and marine ecosystems. A strain of Phormidium-like cyanobacteria has been recently isolated as a free-living epiphyte of leaves of Posidonia oceanica (L.) Delile in the Mediterranean sea and its biology and ecology are herein investigated. It was identified as Halomicronema metazoicum, previously known uniquely as a symbiont of marine sponges. We cultivated it in a range of light irradiances, temperatures and salinities, to establish the most suitable conditions for the production of allelopathic and toxic compounds. The bioactivity of its spent culture medium was measured by means of standard toxicity tests performed on two model organisms. Our results indicate that at least two bioactive compounds are produced, at low and high irradiance levels and at two temperatures. The main compounds influencing the survival of model organisms are produced at the highest temperature and high or intermediate irradiance levels. The present research contributes to the understanding of critical toxigenic relationships among cyanobacteria and invertebrates, possibly influencing the ecology of such a complex environment as P. oceanica. Future isolation, identification and production of bioactive compounds will permit their exploitation for biotechnologies in the field of ecological conservation and medical applications.

Physiological effects of algal metabolites is a key step for the isolation of interesting bioactive compounds. Invertebrate grazers may be fed on live diatoms or dried, pelletized, and added to compound feeds. Any method may reveal some shortcomings, due to the leaking of wound-activated compounds in the water prior to ingestion. For this reason, encapsulation may represent an important step of bioassay-guided fractionation, because it may assure timely preservation of the active compounds. Here we test the effects of the inclusion in alginate (biocompatible and non-toxic delivery system) matrices to produce beads containing two benthic diatoms for sea urchin Paracentrotus lividus feeding. In particular, we compared the effects of a diatom whose influence on P. lividus was known (Nanofrustulum shiloi) and those of a diatom suspected to be harmful to marine invertebrates, because it is often present in blooms (Striatella unipunctata). Dried N. shiloi and S. unipunctata were offered for one month after encapsulation in alginate hydrogel beads and the larvae produced by sea urchins were checked for viability and malformations. The results indicated that N. shiloi, already known for its toxigenic effects on sea urchin larvae, fully conserved its activity after inclusion in alginate beads. On the whole, benthic diatoms affected the embryogenesis of P. lividus, altering the expression of several genes involved in stress response, development, skeletogenesis and detoxification processes. Interactomic analysis suggested that both diatoms activated a similar stress response pathway, through the up-regulation of hsp60, hsp70, NF-κB, 14-3-3 ε and MDR1 genes. This research also demonstrates that the inclusion in alginate beads may represent a feasible technique to isolate diatom-derived bioactive compounds.

Organisms adaptable to extreme conditions share the ability to establish protective biofilms or secrete defence toxins. The extracellular substances that are secreted may contain monosaccharides and other toxic compounds, but environmental conditions influence biofilm characteristics. Microorganisms that are present in the same environment achieve similar compositions, regardless of their phylogenetic relationships. Alternatively, cyanobacteria phylogenetically related may live in different environments, but we ignore if their physiological answers may be similar. To test this hypothesis, two strains of cyanobacteria that were both ascribed to the genus Halomicronema were isolated. H. metazoicum was isolated in marine waters off the island of Ischia (Bay of Naples, Italy), free living on leaves of Posidonia oceanica. Halomicronema sp. was isolated in adjacent thermal waters. Thus, two congeneric species adapted to different environments but diffused in the same area were polyphasically characterized by microscopy, molecular, and toxicity analyses. A variable pattern of toxicity was exhibited, in accordance with the constraints imposed by the host environments. Cyanobacteria adapted to extreme environments of thermal waters face a few competitors and exhibit a low toxicity; in contrast, congeneric strains that have adapted to stable and complex environments as seagrass meadows compete with several organisms for space and resources, and they produce toxic compounds that are constitutively secreted in the surrounding waters.