An Integrated Monitoring Approach to the Evaluation of the Environmental Impact of an Inshore Mariculture Plant (Mar Grande of Taranto, Ionian Sea).

The results of an ex-ante survey aiming to assess the impact of a fish farm in the Mar Grande of Taranto (southern Italy, Mediterranean Sea) on the surrounding environment are reported. There, the implementation of an innovative IMTA plant was planned, with the goals of environment bioremediation and commercially exploitable biomass production. Analyses were conducted in February and July 2018. Both seawater and sediments were sampled at the four corners of the fish farm to detect the existing biological and physico-chemical features. The investigation was performed to identify the best area of the farming plant for positioning the bioremediating system, but also to obtain a data baseline, to compare to the environmental status after the bioremediating action. Data were also analyzed by canonical analysis of principal coordinates (CAP). All the measurements, in particular, microbiology and macrobenthic community characterization using AZTI's Marine Biotic Index (AMBI) and the Multivariate-AMBI (M-AMBI) indices, suggest that the effect of fish farm waste was concentrated and limited to a small portion of the investigated area in relation to the direction of the main current. A site named A3, which was found to be the most impacted by the aquaculture activities, especially during the summer season, was chosen to place the bioremediation system.

Filtration of the Microalga Amphidinium carterae by the Polychaetes Sabella spallanzanii and Branchiomma luctuosum: A New Tool for the Control of Harmful Algal Blooms?

Harmful algal blooms (HABs) are extreme biological events representing a major issue in marine, brackish, and freshwater systems worldwide. Their proliferation is certainly a problem from both ecological and socioeconomic contexts, as harmful algae can affect human health and activities, the marine ecosystem functioning, and the economy of coastal areas. Once HABs establish, valuable and environmentally friendly control actions are needed to reduce their negative impacts. In this study, the influence exerted by the filter-feeding activity of the two sabellid polychaetes Branchiomma luctuosum (Grube) and Sabella spallanzanii (Gmelin) on a harmful dinoflagellate was investigated. Clearance rates (C) and retention efficiencies were estimated by employing the microalga Amphidinium carterae Hulburt. The Cmax was 1.15 ± 0.204 L h-1 g-1 DW for B. luctuosum and 0.936 ± 0.151 L h-1 g-1 DW for S. spallanzanii. The retention efficiency was 72% for B. luctuosum and 68% for S. spallanzanii. Maximum retention was recorded after 30 min for both species. The obtained results contribute to the knowledge of the two polychaetes' filtration activity and to characterize the filtration process on harmful microalgae in light of the protection of water resources and human health. Both species, indeed, were extremely efficient in removing A. carterae from seawater, thus suggesting their employment as a new tool in mitigation technologies for the control of harmful algae in marine environments, as well as in the aquaculture facilities where HABs are one of the most critical threats.

The Pandora's box: Morphological diversity within the genus emAmphiglena/em Claparède, 1864 (Sabellidae, Annelida) in the Mediterranean Sea, with description of nine new species.

We report the description of nine new taxa of sabellid polychaetes belonging to the genus Amphiglena, of which diversity in the Mediterranean Sea has been widely underestimated. Examined material derived from both new collections along the Italian coast, including four CO2 vents/hydrothermal systems, and from a re-examination of older material previously attributed to A. mediterranera (Leydig, 1851) which was so far the only species of the genus reported for the Mediterranean area. The analysis revealed the presence of different taxa also consistent with a previous molecular analysis conducted on material from the Gulf of Naples and the Salento coast (Ionian Sea). This led to an increase in the number of species in the genus and to highlight the occurrence in the Mediterranean Sea of a high diversity within the genus. A key to the Mediterranean Sea species of Amphiglena is also provided. Some taxa, however, remain for the moment undescribed due to the poor preservation of the old material, and the lack of the type material for this taxon. A major revision of all the Mediterranean material previously attributed to A. mediterranea from both morphological and molecular points of view is needed.

Integrated Multitrophic Aquaculture By-Products with Added Value: The Polychaete Sabella spallanzanii and the Seaweed Chaetomorpha linum as Potential Dietary Ingredients.

Aquaculture expansion is limited by the negative environmental impact of the waste and the need for alternative sources in the diet of reared fish. In this framework, for the first time, the survival rates, biomass gain, and fatty acid profiles of the polychaete Sabella spallanzanii and the macroalga Chaetomorpha linum, reared/cultivated as bioremediators in an integrated multitrophic aquaculture system (IMTA), were evaluated for their potential reuse applications. Results showed that these organisms represent a natural source of omega-3 and omega-6. On account of the overall results and the high biomass obtained as by-products, a preliminary study was performed employing both S. spallanzanii and C. linum as new dietary ingredients to feed different sized Dicentrarchus labrax. Fish survival rate, biomass growth, and specific growth rate were determined resulting in no significant differences between control and treated fishes. Histological analyses showed no alterations of the stomach tunica mucosa and submucosa in treated fishes. The eco-friendly approaches applied in the here-realized IMTA system could guarantee the achievement of sustainable by-products represented by the bioremediators S. spallanzanii and C. linum, as well as their reliability as a natural source of compounds beneficial to fish and human health.

First Insight on the Mucus of the Annelid Myxicola infundibulum (Polychaeta, Sabellidae) as a Potential Prospect for Drug Discovery.

Many marine organisms, including invertebrates, produce mucosal matrices having different functions. Besides mechanical protection, the mucus of many invertebrates contains specific compounds to make the animal poisonous and/or distasteful or irritating. The presence of antibiotic molecules is more advantageous for some invertebrates to contrast bacterial attack. In the present study we investigated the mucus of the Mediterranean annelid species Myxicola infundibulum living in a gelatinous envelope made up of dense mucus. Antimicrobial lysozyme-like and antioxidant activities were investigated to highlight the potential interest of the worm mucus as a source of bioactive compounds for biotechnological applications. In order to understand which kind of compounds could be responsible for the detected activities, the mucus of M. infundibulum was chemically characterized in terms of elemental composition, protein, lipid and carbohydrate content. Further chemical characterization was achieved by the advanced analytical technique of multinuclear and multidimensional NMR spectroscopy. NMR spectroscopy revealed the scarcity of lipids which preferentially resulted of alcoholic origin, or otherwise hydroxylate and several aminoacids (valine, leucine and alanine) in the aqueous extract in relation to the protein nature of M. infundibulum mucus. The mucus indeed is mainly composed by water (94% ± 0.7%) whereas its dry weight is made of proteins (36% ± 2.3%) followed by lipids (2.9% ± 0.07%) and carbohydrates (2% ± 0.31%). The mucus exerted a natural antibacterial lysozyme-like activity corresponding to 1.14 mg mL-1 of hen egg-white lysozyme and an antioxidant activity corresponding to 483.00 ± 79.22 nmolTE (Trolox equivalent)/mL sample as Trolox Equivalent Antioxidant Capacity (TEAC) and 276.26 ± 50.76 nmolTE/mL sample as Oxygen Radical Absorbance Capacity (ORAC). Therefore, our findings have potential implications due to the ongoing explosion of antibiotic resistant infections and the need to discover antibacterial agents. Additionally, the observed antioxidant activity is intriguing taking into account the need to find natural antioxidants useful for human health.

Heavy metals in five Sabellidae species (Annelida, Polychaeta): ecological implications.

The present work analyzed three hard-bottom and two soft-bottom species of sabellid polychaetes to determine the content of several heavy metals in their branchial crown and body. The highest concentrations of heavy metals were recorded in the hard-bottom species Branchiomma bairdi, a recent Mediterranean introduction. Differences in the metal concentrations were most notable in the high trace metal levels of the branchial crown for all the studied species. Statistical analysis showed that the Mediterranean hard-bottom species were similar each other in their heavy metal content in the body as well as in the branchial crown and appeared separated from all the other species. Arsenic and vanadium hyperaccumulation in the branchial crowns of the considered sabellid species probably acts as a deterrent for predation. The observed differences among the examined species were discussed not only at the light of habitat colonization but also in terms of the phylogeny.

Filtering activity on a pure culture of Vibrio alginolyticus by the solitary ascidian Styela plicata and the colonial ascidian Polyandrocarpa zorritensis: a potential service to improve microbiological seawater quality economically.

We investigated and compared, by laboratory experiments, the filter-feeding activity on bacteria by the solitary ascidian Styela plicata and the colonial ascidian Polyandrocarpa zorritensis. Clearance rates and retention efficiencies were estimated by using, as only food source, the bacterial species Vibrio alginolyticus selected on account of its importance in aquaculture pathogenicity. The Cmax was 1.4±0.17Lh-1g-1 DW for S. plicata and 1.745Lh-1g-1 DW for P. zorritensis. The highest retention efficiency was 41% corresponding to a removed bacterial biomass of 16.34+1.71 µgCL-1g-1 DW for P. zorritensis and 81% corresponding to a bacterial biomass of 32.28+2.15 µgCL-1g-1 DW for S. plicata. Styela plicata resulted higher efficient than P. zorritensis in removing V. alginolyticus from seawater in experimental tanks, thus representing a more suitable biofilter to restore the quality of microbiologically contaminated waters including those where aquaculture is practiced. Present laboratory experiments represent the first contribution to the comparison of the filtration activity of the two ascidians, as well as to characterize the filtration process on bacterioplankton and pone the basis for future field works aimed to restore bacteriological polluted seawater.

The co-occurrence of the demosponge Hymeniacidon perlevis and the edible mussel Mytilus galloprovincialis as a new tool for bacterial load mitigation in aquaculture.

Pollutants in marine coastal areas are mainly a consequence of anthropogenic inputs, and microorganisms often play a major role in determining the extent of this pollution. Thus, practical and eco-friendly techniques are urgently required in order to control or minimise the pathogenic bacterial problem. The bacterial accumulation of Mytilus galloprovincialis (Lamarck 1919) in the presence or absence of another filter feeder, the demosponge Hymeniacidon perlevis (Montagu 1818) on sewage flowing into the Northern Ionian Sea has been estimated in a laboratory study. On account of the interesting results obtained, we also evaluated the bioremediation capability of the sponges when reared in co-culture with mussels. Specimens of M. galloprovincialis and H. perlevis were collected from the Mar Grande and from the Second Inlet of the Mar Piccolo of Taranto (Northern Ionian Sea, Italy), respectively. In the laboratory, we detected the bacterial abundances in the sewage, in sponge homogenates (both sponges alone and sponges that have been added to sewage with mussels) and in mussel homogenates (both mussels alone and mussels that have been added to sewage with sponges). In the field, we estimated the bacterial concentration in both the seawater within the mussels culture and the seawater collected where mussels were reared in co-culture with sponges. The bacteriological analyses were performed analysing the following parameters: the density of culturable heterotrophic bacteria by spread plate on marine agar, total culturable bacteria at 37 °C on plate count agar and vibrios on thiosulphate-citrate-bile-sucrose-salt (TCBS) agar. Total coliforms, Escherichia coli and intestinal streptococci concentrations were detected by the MPN method. The study demonstrates a higher efficiency of the sponges in removing all the considered bacterial groups compared to the mussels. Due to the conspicuous bacterial accumulation by the sponge, we can conclude that the co-occurrence of the filter-feeder H. perlevis with M. galloprovincialis is a powerful tool in reducing the bacterial load in shellfish culture areas thus playing a role in mitigating the health hazard related to the consumption of edible mussels.

The Mediterranean non-indigenous ascidian Polyandrocarpa zorritensis: Microbiological accumulation capability and environmental implications.

We investigated the bacterial accumulation and digestion capability of Polyandrocarpa zorritensis, a non-indigenous colonial ascidian originally described in Peru and later found in the Mediterranean. Microbiological analyses were carried out on homogenates from "unstarved" and "starved" ascidians and seawater from the same sampling site (Adriatic Sea, Italy). Culturable heterotrophic bacteria (22 °C), total culturable bacteria (37 °C) and vibrios abundances were determined on Marine Agar 2216, Plate Count Agar and TCBS Agar, respectively. Microbial pollution indicators were measured by the most probable number method. All the examined microbiological groups were accumulated by ascidians but differently digested. An interesting outcome is the capability of P. zorritensis to digest allochthonous microorganisms such as coliforms as well as culturable bacteria at 37 °C, counteracting the effects of microbial pollution. Thus, the potential exploitation of these filter feeders to restore polluted seawater should be taken into consideration in the management of this alien species.

Microbiological accumulation by the Mediterranean invasive alien species Branchiomma bairdi (Annelida, Sabellidae): potential tool for bioremediation.

We examined the bacterial accumulation and digestion in the alien polychaete Branchiomma bairdi. Microbiological analyses were performed on worm homogenates from "unstarved" and "starved" individuals and on seawater from the same sampling site (Ionian Sea, Italy). Densities of culturable heterotrophic bacteria (22 °C), total culturable bacteria (37 °C) and vibrios were measured on Marine Agar 2216, Plate Count Agar and TCBS Agar, respectively. Microbial pollution indicators were determined by the most probable number method. B. bairdi was able to accumulate all the six considered microbiological groups which, however, differ in their resistance to digestion. B. bairdi results more efficient than the other two co-occurring sabellids in removing bacteria suggesting that it may counteract the effects of microbial pollution playing a potential role for in situ bioremediation. Thus a potential risk, such as the invasion of an alien species, could be transformed into a benefit with high potential commercial gain and economic feasibility.

Managing the marine aquarium trade: revealing the data gaps using ornamental polychaetes.

The marine aquarium industry has great potential to generate jobs in low-income coastal communities creating incentives for the maintenance of a healthy coral reef, if effectively managed. In the absence of current monitoring or legislation to govern the trade, baseline information regarding the species, number and source location of animals traded is missing despite being critical for its successful management and sustainability. An industry assessment to establish the number and provenance of species of ornamental polychaetes (sabellids and serpulids) traded was undertaken across UK wholesalers and retailers. Six geographical regions exporting fan worms were identified. Singapore contributed the highest percentage of imports, but of only one worm "type" whereas Bali, the second largest source, supplied five different worm "types". Over 50% of UK retailers were supplied by one wholesaler while the remainder were stocked by a mixture of one other wholesaler and/or direct imports from the source country. We estimate that up to 18,500 ornamental polychaetes (16,980 sabellids and 1,018 serpulids) are sold annually in the UK revealing a drastic underestimation of currently accepted trade figures. Incorrect identification (based on exporting region or visual characteristics) of traded animals exacerbates the inaccuracy in market quantification, although identification of preserved sabellids using published keys proved just as inconclusive with high within-species variability and the potential for new or cryptic species. A re-description of the polychaete groups traded using a combination of molecular and morphological techniques is necessary for effective identification and market quantification. This study provides the first assessment of ornamental polychaetes but more importantly highlights the issues surrounding the collection of baseline information necessary to manage the aquarium trade. We recommend that future management should be community based and site-specific with financial and educational support from NGOs, local governments and industry members.

Bioremediation of bacteria in aquaculture waste using the polychaete Sabella spallanzanii.

The excessive release of bacterial pathogens from animal waste into the aquaculture environment has become a major concern for the aquaculture industry. The biological filtration by macroinvertebrates contributes to water purification as a result of the bacterioplankton removal from the water. The filter-feeder polychaete Sabella spallanzanii is known for its ability to accumulate bacteria from the marine environment. In the present study we evaluated the survival, growth and capability of this species to remove several bacterial groups from aquaculture waste in order to ascertain its employment as bioremediator in a farming scenario coupled with the conversion of the wastes into polychaete protein-rich biomass of potentially marketable value. In comparison to other technologies, the employment of S. spallanzanii in waste treatment represents a more attractive option to reduce bacterial loads.

Effects of offshore platforms on soft-bottom macro-benthic assemblages: a case study in a Mediterranean gas field.

The exploitation of fossil fuels in the Mediterranean Sea will likely lead to an increase in the number of offshore platforms, a recognized threat for marine biodiversity. To date, in this basin, few attempts have been made to assess the impact of offshore gas and oil platforms on the biodiversity of benthic assemblages. Here, we adopted a structured experimental design coupled with high taxonomic resolution to outline putative effects of gas platforms on soft-bottom macrofauna assemblages in the North Ionian Sea. The analysis was based on a total of 20,295 specimens of 405 taxa, almost entirely identified at species level. Multivariate and univariate analyses showed idiosyncratic patterns of assemblage change with increasing distance from the platforms. Potential reasons underlying such inconsistency are analyzed and the view that structured experimental monitoring is a crucial tool to quantify the extent and magnitude of potential threats and to provide sound baseline information on biodiversity patterns is supported.

Evaluation of microbiological accumulation capability of the commercial sponge Spongia officinalis var. adriatica (Schmidt) (Porifera, Demospongiae).

This study was carried out to evaluate the microbiological accumulation capability of the demosponge Spongia officinalis var. adriatica. Six microbiological parameters were researched in two sampling periods in the water and in reared sponge samples coming from sites with different degrees of microbial contamination: an off-shore fish farm displaced off the Apulian coast (Southern Adriatic Sea) and a no-impacted area displaced into the Marine Protected Area of Porto Cesareo (Apulian coast-Ionian Sea). We detected the density of culturable heterotrophic bacteria by spread plate on marine agar, total culturable bacteria at 37 degrees C on Plate Count Agar and vibrios on thiosulphate-citrate-bile-sucrose-salt (TCBS) agar. Total and fecal coliforms as well as fecal streptococci concentrations were detected by the MPN method. Bacterial densities were always higher in the sponge homogenates compared with the corresponding seawater in the sampling points and in both sampling periods. As regard vibrios, total culturable bacteria at 37 degrees C and fecal streptococci concentrations, the highest values were observed in the sponge samples coming from the off-shore fish farm during the summer period. The ability of Spongia officinalis var. adriatica to accumulate the microbial pollution indicators suggests that this species can be employed as a bioindicator for monitoring water quality.

Filter-feeder macroinvertebrates as key players in culturable bacteria biodiversity control: a case of study with Sabella spallanzanii (Polychaeta: Sabellidae).

The present study investigates the effect of the filtering activity of Sabella spallanzanii on the culturable heterotrophic bacterial community through the comparison of the bacterial diversity in transplanted polychaetes and the surrounding seawater. For isolation of culturable heterotrophic bacteria, seawater samples as well as polychaete homogenates were plated in triplicates onto Bacto Marine Agar 2216 (Difco). All the colonies grown were isolated, subcultured and identified by several morphological, biochemical and cultural methods. Some bacterial genera showed higher average abundances in polychaetes than in seawater (i.e. Lucibacterium and Photobacterium). Aeromonas represented a conspicuous component of the bacterial community both in S. spallanzanii and seawater. The presence of Cytophaga and Pseudomonas was also relevant in the examined seawater samples. The selective concentrations of some bacterial genera inside S. spallanzanii either by grazing on bacteria or their capability as bacterial reservoirs, provides evidence for the role of macrobenthic invertebrates as key determinants for microbial diversity.

Bacterial accumulation by Branchiomma luctuosum (Annelida: Polychaeta): a tool for biomonitoring marine systems and restoring polluted waters.

In this study, we examined the bacterial accumulation in the filter feeder polychaete Branchiomma luctuosumGrube (Sabellidae). Analyses were performed on worm homogenates from 'unstarved' and 'starved' individuals, and seawater from the same sampling site (Gulf of Taranto, Western Mediterranean, Italy). Densities of culturable heterotrophic bacteria (22 degrees C), total culturable bacteria at 37 degrees C and halophilic vibrios at 22 and 35 degrees C were measured on Marine Agar 2216, Plate Count Agar and thiosulphate-citrate-bile-salt-agar (TCBS) plus 2% NaCl, respectively. Total and faecal coliforms as well as faecal streptococci were determined by the Most Probable Number method. Results showed that B. luctuosum is able to accumulate all the considered six microbiological groups with a higher efficiency for autochthonous bacteria. The analysis also indicated that bacterial groups differ in their resistance to digestion by B. luctuosum. Our data suggest the potential role of B. luctuosum as an useful bioindicator to assess low levels of microbiological water pollution as well as bioremediator of microbial polluted waters.

Filtering activity of Spongia officinalis var. adriatica (Schmidt) (Porifera, Demospongiae) on bacterioplankton: implications for bioremediation of polluted seawater.

A study on the filtering activity has been carried out on reared specimens of the demosponge Spongia officinalis var. adriatica coming from an off-shore farm displaced off the Apulian coast (Ionian Sea). The experience was carried out under laboratory conditions, by using natural seawater collected from the sponge environment. The study demonstrates a high efficiency of the sponge in removing bacteria. Bacterial concentration significantly decreases in presence of the sponge, with a marked drop after 2 h from the start of the experience. The maximum clearance rate was 210 ml h(-1) g(-1) DW at 60 min. Retention efficiency reached the highest value of 61% at 120 min. The bacterial density removed by the S. officinalis filtering activity was 12.3 +/- 1.8 x 10(4) cells ml(-1) corresponding to a biomass of about 11.7 +/- 1.4 microg Cl(-1). The sponge fed preferentially large- and medium-size bacteria, whereas the small ones are fed after the removal of the largest size categories. The results obtained suggest that S. offcinalis is a suitable species for marine environmental bioremediation.

Sabella spallanzanii filter-feeding on bacterial community: ecological implications and applications.

The filtration process of Sabella spallanzanii Gmelin on bacterial community was studied in a coastal area of the Northern Ionian Sea (Mediterranean Sea) at three sites, S. Vito, Lido Gandoli and Lido Silvana, where some specimens of S. spallanzanii were transplanted. Analyses were performed both on water and worm samples. A total of six microbial groups were examined: culturable heterotrophic bacteria, total culturable bacteria at 37 degrees C, culturable vibrios, total and fecal coliforms and fecal streptococci. The bacterial densities were usually orders of magnitude higher in the worm homogenates than in the corresponding seawater and the highest values were observed in August. The ability of S. spallanzanii to accumulate the microbial pollution indicators suggests this species can be employed as a bioindicator for monitoring water quality. Moreover, the accumulation capability of S. spallanzanii for specific micro-organisms provides a potential role in sewage bioremediation.

Polychaetes as environmental indicators revisited.

The utilization of polychaetes in descriptive ecology is reviewed in the light of recent research especially concerning the biota hard bottom environments. Polychaetes, often linked in the past to the concept of opportunistic species able to proliferate after an increase in organic matter, have played an important role especially with regard to impacted soft-bottom habitats. Increased knowledge of the group, suggests that not only opportunistic species can be utilised as indicators, so that these organisms can be disengaged from the old concept of opportunistic taxa. Moreover, recent researches conducted on this group allowed demonstrating as surrogacy is not always applicable. Among polychaetes inhabiting hard bottom environment, the analysis of family Syllidae appears particularly promising. Studied conducted in our laboratory demonstrated as syllid species decrease in abundance or completely disappear under varying sources of negative impact. The distribution of species also appeared indicative in underlying effects of marine protected areas (MPA) functioning, or in describing different climatic areas within biogeographical sectors. It is obvious that good results can only be obtained on the basis of good taxonomic resolution. We suggested that, in monitoring studies, operational time could be optimized not only by working at a higher-level on the whole invertebrate data set, but by also selecting a particularly indicative group and working at fine level.

Clearance rates of Sabella spallanzanii and Branchiomma luctuosum (Annelida: Polychaeta) on a pure culture of Vibrio alginolyticus.

The influence exerted by filter-feeding activity on bacterial density by two sabellid species from the Mediterranean Sea (Ionian Sea, Italy), Branchiomma luctuosum Grube and Sabella spallanzanii Gmelin (Annelida: Polychaeta) was investigated. Clearance rates and retention efficiencies were estimated utilizing the species Vibrio alginolyticus selected on account of previous field studies and its importance in fish culture pathogenicity. The Cmax was 43.2+/-2.63 L h(-1) g(-1) DW for B. luctuosum and 12.4+/-2.22 L h(-1) g(-1) DW for S. spallanzanii. The Retention efficiency was 98% corresponding to a removed bacterial biomass of 44.8+7.88 microgC L(-1) g(-1) DW for B. luctuosum and 70% corresponding to a bacterial biomass of 23.8+2.95 microgC L(-1) g(-1) DW for S. spallanzanii. Maximum retention was recorded after 20 min for the first species and after 30 min for the second one. Present laboratory experiments represent a contribution to the knowledge of the filtration activity of the two polychaetes, characterizing the filtration process on bacterioplankton. Both species resulted extremely efficient in removing V. alginolyticus from seawater in experimental tanks, thus confirming the previous data from the field studies and suggesting their employment as biofilters of microbially contaminated waters in intensive aquaculture.