Arnaldo Marín

Dynamics of a peculiar plant-herbivore relationship: the photosynthetic ascoglossan Elysia timida and the chlorophycean Acetabularia acetabulum

The ascoglossan mollusc Elysia timida Risso, 1818 retains functional chloroplasts from its algal food, the chlorophycean Acetabularia acetabulum (L.). Photosynthates from the plastids are an important source of organic nutrients for the mollusc. Chloroplast exploitation has an ecological function, allowing the ascoglossan to live entirely on an algal diet which is of limited, seasonal availability to other herbivores. Between October 1987 and July 1988, the annual evolution of the molluscan and algal populations was studied in a cove of Mazarrón Bay, southeast Spain. The population density of the mollusc is highly dependent on its food supply, being controlled by the seasonal life cycle of the algal population. During its life cycle, the degree of grazing by the mollusc decreases with increasing algal calcification, the cell walls of the alga progressively calcify, and the eventually highly calcificied stalks are completely resistant to ascoglossan grazing. In contrast, the exploitation of the algal chloroplasts retained by the molluscs increases during the seasonal cycle. The progressively increasing scarcity of food during the seasonal cycle may have led to the retention of symbiotic chloroplasts by E. timida. The developmental strategy of the ascoglossan also changes during the year: when food is abundant (in November, December, January, February and March) it is direct, with no planktonic larval phase, when food is scarce (in October, April, May and June) it is lecithotrophic, with a short planktonic larval phase. Chloroplast retention acts as a buffer, alleviating the effects of annual changes in density, structure and abundance of the alga on the nutritional state of the molluse.

Defensive relationships between Caulerpa prolifera and its shelled sacoglossan predators

Abstract The prey-predator relationship between the green alga Caulerpa prolifera and three shelled sacoglossans, Oxynoe olivaceca, Lobiger serradifalci and Cylindrobulla fragilis , have been investigated in order to clarify the role of the chemical molecules in the defense of both alga and molluscs. Caulerpa prolifera contains as main component, among the secondary metabolites, caulerpenyne (1), a small molecule displaying a sesquiterpenoid skeleton. A defensive role, due to the presence of a protected 1,4-dialdehyde, was previously suggested for caulerpenyne (1). In spite of this protection, three sacoglossans, the infaunal C. fragilis and the epifaunal L. serradifalci and O. olivacea , live closely associated with Caulerpa prolifera . The Thyrrhenian O. olivacea is able to modify the algal metabolite into a couple of more toxic compounds, oxytoxin-1 (2) and -2 (3), characterized by the presence of aldehyde groups. The same defensive strategy is adopted by populations of O. olivacea from Spanish coasts and by populations of C. Fragilis and L. serradifalci from S W Italy. The biotransformation of caulerpenyne (1) leads first to oxytoxin-1 (2), a monoaldehyde, which is compartmentalized into the parapodia of L. serradifalci and into the tail of O. olivacea and then, in O. olivacea and C. fragilis , to oxytoxin-2 (3) which displays the 1,4-conjugated dialdehyde partial moiety, present also in the structure of the potent antifeedant polygodial (4). The presence of the organic molecules has been detected by chemical methods, in particular by NMR (Nuclear Magnetic Resonance) of the extracts of the fresh tissues of the molluscs with deuterated solvents, avoiding any Chromatographie step and chemical treatment.

Transfer of secondary metabolites from the spongesDysidea fragilis andPleraplysilla spinifera to the mantle dermal formations (MDFs) of the mudibranchHypserlodoris webbi

The opisthobranch molluscHypselodoris webbi is able to select, among its potential preys, sponges chemically rich in furanosesquiterpenoids. The sequestered secondary metabolites act as defensive allomones against predators and are accumulated in some dorsal glands (MDFs). This transfer from sponges to MDFs has been proven by maintainingH. webbi together with some selected sponges in an aquarium for a prolonged period.

Relationship between sedimentation rates and benthic impact on Maërl beds derived from fish farming in the Mediterranean

The aim of this work was to study the dispersion of particulate wastes derived from marine fish farming and correlate the data with the impact on the seabed. Carbon and nutrients were correlated with the physico-chemical parameters of the sediment and the benthic community structure. The sedimentation rates in the benthic system were 1.09, 0.09 and 0.13 g m⁻² day⁻¹ for particulate organic carbon (POC), particulate organic nitrogen (PON) and total phosphorus (TP), respectively. TP was a reliable parameter for establishing the spatial extent of the fish farm particulate wastes. Fish farming was seen to influence not only physico-chemical and biological parameters but also the functioning of the ecosystem from a trophic point of view, particularly affecting the grazers and the balance among the trophic groups. POC, PON and TP sedimentation dynamics reflected the physico-chemical status of the sediment along the distance gradient studied, while their impact on the benthic community extended further. Therefore, the level of fish farm impact on the benthic community might be underestimated if it is assessed by merely taking into account data obtained from waste dispersion rates. The benthic habitat beneath the fish farm, Maërl bed, was seen to be very sensitive to aquaculture impact compared with other unvegetated benthic habitats, with an estimated POC-carrying capacity to maintain current diversity of 0.087 g C m⁻² day⁻¹ (only 36% greater than the basal POC input). Environmental protection agencies should define different aquaculture waste load thresholds for different benthic communities affected by finfish farming, according to their particular degree of sensitivity, in order to maintain natural ecosystem functions.

Effect of temperature on biogeochemistry of marine organic‐enriched systems: implications in a global warming scenario

Coastal biogeochemical cycles are expected to be affected by global warming. By means of a mesocosm experiment, the effect of increased water temperature on the biogeochemical cycles of coastal sediments affected by organic-matter enrichment was tested, focusing on the carbon, sulfur, and iron cycles. Nereis diversicolor was used as a model species to simulate macrofaunal bioirrigation activity in natural sediments. Although bioirrigation rates of N. diversicolor were not temperature dependent, temperature did have a major effect on the sediment metabolism. Under organic-enrichment conditions, the increase in sediment metabolism was greater than expected and occurred through the enhancement of anaerobic metabolic pathway rates, mainly sulfate reduction. There was a twofold increase in sediment metabolism and the accumulation of reduced sulfur. The increase in the benthic metabolism was maintained by the supply of electron acceptors through bioirrigation and as a result of the availability of iron in the sediment. As long as the sediment buffering capacity toward sulfides is not surpassed, an increase in temperature might promote the recovery of organic-enriched sediments by decreasing the time for mineralization of excess organic matter.

Secondary metabolites from Mediterranean Elysioidea: origin and biological role

Abstract Three Mediterranean sacoglossans (Elysia translucens, Bosellia mimetica and Elysia viridis) have been submitted to chemical investigations. The chemical structure (diterpenoids and polypropionates) of their potential allomones has been clarified. Some metabolites (diterpenoids) are also present in the preferred algal food of the molluscs. E. viridis is able to construct de novo metabolites (polypropionates) absent in the algae. A more general evolutionary trend of Mediterranean Elysioidea molluscs is suggested by adding the results reported herewith to the previous studies on Elysia timida and Thuridilla hopei and by comparing the metabolic pattern of T. hopei with that of the algae Derbesia tenuissima.

Integrative ecotoxicological assessment of sediment in Portmán Bay (southeast Spain).

Portmán Bay, southeast Spain, contains the most seriously metal-contaminated sediments of the Mediterranean Sea. From 1958 to 1991, approximately 50 million tons of mine tailings were dumped into the bay, completely filling up the bay and dispersing over an extensive area of the continental platform and continental slope. The objective of our study was to characterize the nature and extent of metal contamination and the responses of natural communities to it and to assess the toxicity of the sediment deposits 10 years after mining had ceased. We studied the physical and chemical characteristics of the sediments and toxicity (of the porewater and sediment-water interface) using two sea urchin species (Arbacia lixula and Paracentrotus lividus). Metal bioavailability and patterns of macroinvertebrate community composition along the contamination gradient were also studied. Univariate and multivariate analyses showed positive correlation between the sediment metal concentrations associated to the all biological effects (sea urchins toxicity tests and benthic indices). The effects of sediment contamination on the benthic community structure are visible among sampling stations.

Pulse-discharges of mining wastes into a coastal lagoon: Water chemistry and toxicity

Historical mining activities have led to the presence of enormous quantities of mining wastes, which cover large extensions of the Cartagena-La Unión mining district (SE, Spain). In the present work, we study the pulse entrance of mining wastes through two temporary streams (wadis) into the Mar Menor coastal lagoon in two torrential rain events and during a dry period. The characteristics of the runoff pointed to the generation of acid mine drainages in wastes, the acidified stormwater runoff loaded with particulate and dissolved heavy metals causing toxicity in the lagoon waters. The dissolved metals, which mainly affected the sampling stations located close to the wadi discharges, were rapidly eliminated from the water column, whereas the particulate metals were transported further and affected a wider area. Finally, both particulate and dissolved metals are eliminated from the water column and are accumulated in the sediments of the lagoon. The results of the water toxicity tests using sea-urchin embryos indicated that water toxicity disappeared within a few days in the most distant stations, but lasted longer and depended on the climatic conditions of the subsequent days in stations closer to the wadi outlets.

Histological and biochemical bases of defense mechanisms in four species of Polybranchioidea ascoglossan molluscs

The multiple defensive strategies of four Mediterranean ascoglossan molluscan species, belonging to two different Polybranchioidea families, Cyerce cristallina (Trinchese, 1881) and Caliphylla mediterranea (Costa A., 1869) (Polybranchiidae), and Ercolania funerea (Costa A., 1867) and Placida dendritica (Alder and Hancock, 1843) (Stiligeridae), were studied. E. funerea, P. dendritica and C. mediterranea were collected from the lake Fusaro (Arco Felice, Naples) in 1992. C. cristallina was collected from Capo Miseno (Bay of Naples) in 1991. C. cristallina and E. funerea easily undergo autotomy of dorso-lateral appendages (cerata) followed by an extraordinarily quick (8 to 10 d) regeneration of the latter. Histological analyses showed the presence, at the basis of both normal and regenerated cerata of these species, of a muscular sphincter which facilitates the autotomic process. C. mediterranea and P. dendritica, which do not undergo autotomy, lack this anatomical feature but feed upon algae belonging to the Chaetomorpha and Bryopsis genera, and, as shown by electron microscopy studies, retain large quantities of chloroplasts which they use as camouflage amidst algae and to escape predation. E. funerea also exploits this behavioral defense mechanism. Histological investigations also revealed in the cerata of all four species several multi-cellular mucous glands responsible for the secretion of the slime typical for these molluscs. C. cristallina, E. funerea and P. dendritica secrete large amounts of slime, whose extracts displayed ichthyotoxic activity when assayed by the Gambusia affinis ichthyotoxicity assay, while extracts of C. mediterranea slime were not toxic. A chemical analysis of the slime, mantle and cerata led to the isolation of polypropionate α- and γ-pyrones from all species except C. mediterranea. These secondary metabolites possess structures that differ only by the degree of methylation and the geometry of double bonds of the side chain and are specifically distributed in cerata and slime of C. cristallina and E. funerea. The pyrones also display different activities in the Hydra vulgaris regeneration assay and in the G. affinis ichthyotoxicity test and, depending on their structural features and tissue distribution, are likely to play a role either as defense allomones or as supportive inducers of cerata regeneration. In conclusion, combined biological observations and histological and chemical techniques generated valuable information on the defensive behavior of these four ascoglossan species which exploit various combinations of the same behavioral and/or chemical defensive strategies and thus successfully avoid predation.

A manipulative field experiment to evaluate an integrative methodology for assessing sediment pollution in estuarine ecosystems

The assessment of sediment contamination is of crucial importance for the management of estuarine ecosystems. Environmental risk assessment of oil pollution must be specific to these ecosystems because of their unique toxicant bioavailability dynamics, which is not comparable with that of other ecosystems where the environmental parameters are less variable. The goal of this work was to test in two European estuarine areas (Ria de Aveiro, Portugal; La Manga, Spain) whether the common methodology used to evaluate sediment pollution in marine sediment (amphipod toxicity tests and community structure analysis) is suited to these physico-chemically unique systems. Manipulative field experiments were conducted at three oil concentration levels, to compare resulting changes in community structure with laboratory and in situ amphipod toxicity tests carried out with native amphipod species Corophium multisetosum (Atlantic area) and Microdeutopus gryllotalpa (Mediterranean area). The impact of the toxicant was reflected in the community structure and toxicity tests, both of which were correlated with oil concentration. These results point to this methodology being a reliable tool for assessing and monitoring pollution in estuarine areas.