Raffaele D'Adamo

Integrated biomonitoring assessment of the Lesina Lagoon (Southern Adriatic Coast, Italy): preliminary results

An integrated biomonitoring program for marine and coastal ecosystems quality assessment combines the chemical characterization of a site with the evaluation of the possible structure alterations of its living communities. This can be considered an useful tool for better identifying the summarized effects of all the components interacting with the biota. Such an integrated procedure was carried out for the assessment of the quality of the Lesina Lagoon (Southern Adriatic Coast, Italy). The water parameters levels showed a high primary production (2–6 μg l−1); the sediment and pore water toxicity bioassays recorded a low or moderate diffused toxicity. Besides, the benthic meiofauna community structure was characterized by prevalent Nematoda taxa with a homogeneous spatial distribution. On these basis, the Lesina Lagoon seems to be characterized by a prevalent organic pollution mainly related to agricultural and zootechnical activities which, due to the lagoon’s conformation, presents a homogeneous spatial distribution.

Gametes and embryos of sea urchins (Paracentrotus lividus, Lmk., 1816) reared in confined conditions: their use in toxicity bioassays

Sea urchins of the species Paracentrotus lividus, collected along the Southern Adriatic coast at the end of their reproductive season, were reared in a recirculating aquarium under controlled temperature and feeding conditions in order to induce off-season maturation of gametes to be used in toxicity bioassays. After 14 weeks of rearing, the gonad index was calculated, histological analyses of the gonads were performed, their fertilisation ability was measured and the EC50 for copper in embryotoxicity bioassays was evaluated. The sperm motility pattern (based on the percentages of total motile and rapid sperm, together with sperm velocity parameters) was assessed using a computerised analyser. All specimens showed a significant increase in gonad yield with respect to field-collected animals and were in pre-spawning or spawning stages. The motility parameters, fertilisation ability and EC50 level of the collected gametes were comparable to those of field-matured specimens. In conclusion, the sea urchins reared under experimental conditions were characterised by faster gonad maturation than the field population, producing viable gametes and larvae whose biological response in toxicity bioassays was similar to that of field-reared specimens of P. lividus.

Cryopreserved semen in ecotoxicological bioassays: sensitivity and reliability of cryopreserved Sparus aurata spermatozoa.

The aim of this study was to evaluate the feasibility of using cryopreserved S. aurata semen in spermiotoxicity tests. Cryopreservation is a biotechnology that can provide viable gametes and embryos on demand, rather than only in the spawning season, thus overcoming a limitation that has hindered the use of some species in ecotoxicological bioassays. Firstly, the sperm motility pattern of cryopreserved semen was evaluated after thawing by means of both visual and computer-assisted analyses. Motility parameters in the cryopreserved semen did not change significantly in the first hour after thawing, meaning that they were maintained for long enough to enable their use in spermiotoxicity tests. In the second phase of the research, bioassays were performed, using cadmium as the reference toxicant, in order to evaluate the sensitivity of cryopreserved S. aurata semen to ecotoxicological contamination. The sensitivity of the sperm motility parameters used as endpoints (motility percentages and velocities) proved to be comparable to what has been recorded for the fresh semen of other aquatic species (LOECs from 0.02 to 0.03 mg L(-1)). The test showed good reliability and was found to be rapid and easy to perform, requiring only a small volume of the sample. Moreover, cryopreserved semen is easy to store and transfer and makes it possible to perform bioassays in different sites or at different times with the same batch of semen. The proposed bioassay is therefore a promising starting point for the development of toxicity tests that are increasingly tailored to the needs of ecotoxicology and environmental quality evaluation strategies.

Gamete Maturation and Gonad Growth in Fed and Starved Sea Urchin Paracentrotus lividus (Lamarck, 1816)

ABSTRACT A 4-wk rearing trial of the sea urchin Paracentrotus lividus was carried out in a closed-circuit system in the presence and absence of food supply to evaluate the short-term response of gametogenesis to different feeding conditions. At the end of the trial, the gonad index (GI) was calculated, histological analyses of the gonads were performed, gamete fertilization ability was evaluated, and sperm motility was assessed by computerized motility analysis. Starvation significantly affected gametogenesis, whereas developing gametes were always observed in fed animals, whose GI had doubled by the end of the 4-wk trial. No differences were recorded between gametes from reared (fed) urchins and wild-collected ones. Although spent gonads frequently contained unspawned motile spermatozoa or morphologically intact eggs, the fertilization ability of gametes from starved urchins was significantly lower. It may be concluded that, although they were at the end of the reproductive season, the specimens fed ad libitum were able to reactivate the gamete maturation process immediately. This ability can be used in short-term procedures for roe enhancement and gamete production, particularly for specimens from areas in which environmental conditions determine slow gametogenesis and a consequently short reproductive season.

Melatonin-mediated effects on killifish reproductive axis

The aim of this study was to investigate the melatonin-mediated effects upon the neuroendocrine axis of the brackish killifish (Fundulus heteroclitus), a suitable experimental model to study reproductive events. The ability of melatonin to enhance reproductive capacity (fecundity, embryo survival and hatching rate) inducing the transcriptional activity of gonadotropin releasing hormone (gnrh), luteinizing hormone receptor (lhr) and melatonin receptor (mtnr) was investigated in adult females. Moreover, the melatonin-mediated enhancement of killifish sperm motility and velocity was found consistent with higher fecundity of melatonin-exposed fishes. As a further extent, Fourier Transform Infrared (FT-IR) microspectroscopy evidenced a reduction of lipid unsaturation level on isolated spermatozoa from treated males. Moreover, the reduction of mtnr gene expression during embryo development and lower biometric parameters documented in the larvae from melatonin-exposed parents suggest that melatonin acts as a hormonal mediator able to transfer the environmental signal to oocytes and then to embryos as inheritance of adaptive environmental changes. These results support the positive role of melatonin on killifish reproduction and its role as a maternal factor on embryo and larval development.

Zooplankton responses to hydrological and trophic variability in a Mediterranean coastal ecosystem (Lesina Lagoon, South Adriatic Sea)

Spatial and temporal variability in zooplankton was studied at eight stations located in the Lesina Lagoon (South Adriatic Sea) Salinity, temperature, dissolved oxygen and chlorophyll a (in the lagoon) at these stations were also assessed. The zooplankton community was characterised by clear seasonal oscillations and mostly represented by copepods and their larvae. The dominant copepod species were Calanipeda aquaedulcis and Acartia tonsa, which exhibited spatial–temporal segregation in the lagoon. C. aquaedulcis copepodites seemed to be better adapted to oligotrophic and oligohaline conditions compared with the A. tonsa population. The invasive species A. tonsa has completely replaced the formerly abundant Acartia margalefi. A positive correlation was found between abundances, total species numbers and trophic state. An increasing abundance trend was shown from the lagoon towards the sea. The highest diversity indices were recorded at the two channel inlets, during high tide due to the absence of a clear dominance of one or more coastal species and the co-occurrence of species of lagoon and marine origin.

Motility of cryopreserved spermatozoa for the ecotoxicological evaluation of aquatic environments

A new approach to environmental studies was investigated by the authors, who propose the use of cryopreserved biological systems in ecotoxicological bioassays. The feasibility of spermiotoxicity tests using cryopreserved semen of the sea bream Sparus aurata, with sperm motility parameters as the endpoint, was evaluated. Thawed sperm was incubated in environmental samples (sediment elutriate and dumpsite leachate) and in a reference toxicant (cadmium) at scaled concentrations. Motility was then evaluated by video-microscopy using both visual and computer-assisted analyses. Activation time, sperm motility and velocity and motility duration were assessed on thawing and at the end of the incubation time, and the difference with respect to the control was statistically evaluated. All the endpoints of the bioassay proved to have good sensitivity even at the highest dilutions of the tested matrices. Observed differences in the sensitivity thresholds of the endpoints were considered to be representative of different aspects of sperm physiology. Therefore the proposed bioassay is a promising starting point for the development of toxicity tests that are increasingly tailored to the needs of ecotoxicology and environmental quality evaluation strategies for aquatic environments.

Phytoplankton diversity in Adriatic ports: Lessons from the port baseline survey for the management of harmful algal species

An inventory of phytoplankton diversity in 12 Adriatic ports was performed with the port baseline survey. Particular emphasis was put on the detection of harmful aquatic organisms and pathogens (HAOP) because of their negative impact on ecosystem, human health, and the economy. Phytoplanktonic HAOP are identified as species, either native or non-indigenous (NIS), which can trigger harmful algal blooms (HAB). A list of 691 taxa was prepared, and among them 52 were classified as HAB and five as NIS. Records of toxigenic NIS (Pseudo-nitzschia multistriata, Ostreopsis species including O. cf. ovata) indicate that the intrusion of non-native invasive phytoplankton species has already occurred in some Adriatic ports. The seasonal occurrence and abundance of HAOP offers a solid baseline for a monitoring design in ports in order to prevent ballast water uptake and possible expansion of HAOP outside their native region.

Agar-based Biocomposites slow down Progression in the Reproductive Cycle Facilitating Synchronization of the Gonads of Reared Specimens of Paracentrotus lividus

Background: Biopolymers have many fields of application. In Echiniculture they are usually employed to bind trial diets, although a systematic study on the performances of biopolymers as feed binders is lacking.