Victor Quintino

Benthic biotopes remote sensing using acoustics

Abstract The present work applies novel methodologies to the study of sublittoral benthic biotopes, by combining the information given by underwater acoustic and biological survey techniques. The acoustic seabed classification system QTC VIEW™ was used to map the acoustic diversity between 5 and 40 m water depth on the shelf off “Ria de Aveiro”, Western coast of Portugal. Ground-truth was undertaken using an analysis of superficial sediments grain-size, and compared to the species composition and distribution of macrofaunal communities. Sedimentary and biological data were submitted to ordination analysis, and the acoustic data to both ordination and cluster analysis. The acoustic classes identified were mapped using a geographical information system. The acoustic results showed a very clear geographic pattern, with the acoustic classification being coincident where survey lines crossed, confirming the stability of the classification procedure. At the optimal splitting level, three acoustic classes were obtained. These classes were correlated to differences in coarse, fine and very fine sands. Additional real bottom differences in the grain-size of the coarser sand classes were not detected by the acoustic method, possibly due to the fact that they showed a similar degree of compactness. The benthic ecological data suggests only two main communities, which correspond to the outcomes of a two-class acoustic split. Therefore, a mismatch was noticed between the optimal acoustic split and the number of major biological communities present. However, by dropping the acoustic solution down to two classes, an optimal relationship to benthic communities is achieved. Overall, results suggest that the acoustic system provides very valuable and important data for mapping soft sediment biotopes, even in areas of relative bottom monotony such as the one analysed, but careful ground-truth is required to ensure that the acoustic class splits are biologically relevant.

Looking for suitable biomarkers in benthic macroinvertebrates inhabiting coastal areas with low metal contamination: Comparison between the bivalve Cerastoderma edule and the Polychaete Diopatra neapolitana

Metals accumulated in marine sediments are often a threat to benthic communities. With the recognized importance and wide use of stress biochemical responses as indicators of metal contamination it becomes essential to compare these markers between different species and verify their ubiquity and accuracy. Using wild Diopatra neapolitana and Cerastoderma edule, collected at several areas differing in metal contamination, this study aimed to assess the use of these two macrobenthic species as sentinel organisms and to determine the applicability of currently used biomarkers in benthic species exposed to a range of low metal and As concentrations. Total metal accumulation and intracellular partitioning was analyzed and metal-induced alterations were assessed through the analysis of several biochemical parameters in both organisms, including stress-induced reactive oxygen species (ROS), lipid peroxidation and protein content, the activity of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and of glutathione S-transferase (GST). Metal chelation by Metallothioneins (MTs) was also determined. Results revealed that D. neapolitana accumulated higher levels of metals when compared to C. edule, independently of the sediment concentration. Results also showed strong species-specific responses to metals and differences in the ability to sequester metals. Overall, C. edule showed to be more efficient metal chelator and precipitatior than D. neapolitana, which was less tolerant and presented oxidative stress. MTs proved to be a good predictor of metal accumulation in both species, even under low metal exposures. On the other hand, lipid peroxidation was a good indicator of oxidative damage, only observed in D. neapolitana, which was a result of higher metal retention in the soluble fraction.

Spatial patterns of benthic macroinvertebrates in intertidal areas of a Southern European estuary: the Tagus, Portugal

This study characterizes the composition and spatial distribution patterns of the benthic macrofauna in the intertidal mudflats of the Tagus estuary, western Portugal. A total of 68 species, more than 226,000 specimens with a total wet weight biomass of approximately 1170 g were identified in 380 sites. The species Streblospio shrubsolii, Cyathura carinata, Tharyx sp., Hydrobia ulvae and Tubificids were the most common and abundant. Scrobicularia plana strongly dominated the biomass. The invertebrate macrofauna of the Tagus estuary shows similarities to what is known from other temperate mudflats. The diversity of species, their overall abundance and the ratio of Molluscs plus Crustaceans to Polychaete species corroborate the distinctiveness between temperate and tropical mudflats and sandflats. The spatial distribution of the fauna reflects the sediment characteristics but the relationship between the environmental and the biological data is not as strong as obtained for sublittoral areas. This relationship diminishes from the sublittoral shelf to sublittoral estuarine areas, showing minimum values in this study, suggesting that such a relationship is less straightforward as natural disturbance increases. Nevertheless, a mixture of grain-size, elevation (inundation time) and particular habitats (relic oyster beds) form the best explanatory factors for the spatial distribution patterns of the intertidal benthic macrofauna of the Tagus estuary.

Acoustic seabed classification of marine habitats: studies in the western coastal-shelf area of Portugal

Two single-beam, seabed-classification systems, QTC VIEW Series IV and QTC VIEW Series V, were used to identify and map biosedimentary gradients in a mid-shelf area off Western Portugal. The survey area has a moderate slope, a depth ranging from 30 to 90 m along a 3.5-km axis perpendicular to the shoreline, and is characterized by smooth sedimentary and biological gradients. Ground truth for sediment grain size and macrofaunal communities was based on grab sampling at 20 sites. The sedimentary and biological data were analysed using classification and ordination techniques. The acoustic data were analysed with qtc impact software and classified into acoustic classes. The affinity groups obtained in each data set were mapped using a Geographics Information System. All showed good agreement and identified prevailing gradients along a northwest-southeast direction. Three acoustic classes were identified, corresponding to the predominant sediment types, namely fine sand with low silt and clay content, silty, very fine sand, and mud. A close relationship with benthic communities was also verified, although less marked because benthic communities continuously change along the northwest-southeast gradient. Overall, the acoustic system coupled with ground-truthing data was able to discriminate and characterize the various benthic biotopes in the survey area.

Molecular analysis of bacterial communities and detection of potential pathogens in a recirculating aquaculture system for Scophthalmus maximus and Solea senegalensis.

The present study combined a DGGE and barcoded 16S rRNA pyrosequencing approach to assess bacterial composition in the water of a recirculating aquaculture system (RAS) with a shallow raceway system (SRS) for turbot (Scophthalmus maximus) and sole (Solea senegalensis). Barcoded pyrosequencing results were also used to determine the potential pathogen load in the RAS studied. Samples were collected from the water supply pipeline (Sup), fish production tanks (Pro), sedimentation filter (Sed), biofilter tank (Bio), and protein skimmer (Ozo; also used as an ozone reaction chamber) of twin RAS operating in parallel (one for each fish species). Our results revealed pronounced differences in bacterial community composition between turbot and sole RAS, suggesting that in the systems studied there is a strong species-specific effect on water bacterial communities. Proteobacteria was the most abundant phylum in the water supply and all RAS compartments. Other important taxonomic groups included the phylum Bacteriodetes. The saltwater supplied displayed a markedly lower richness and appeared to have very little influence on bacterial composition. The following potentially pathogenic species were detected: Photobacterium damselae in turbot (all compartments), Tenacibaculum discolor in turbot and sole (all compartments), Tenacibaculum soleae in turbot (all compartments) and sole (Pro, Sed and Bio), and Serratia marcescens in turbot (Sup, Sed, Bio and Ozo) and sole (only Sed) RAS. Despite the presence of these pathogens, no symptomatic fish were observed. Although we were able to identify potential pathogens, this approach should be employed with caution when monitoring aquaculture systems, as the required phylogenetic resolution for reliable identification of pathogens may not always be possible to achieve when employing 16S rRNA gene fragments.

Health concerns of consuming cockles (Cerastoderma edule L.) from a low contaminated coastal system.

Commercial and recreational harvesting of shellfish within the coastal systems is usually very extensive. Since these ecosystems are frequently subjected to contamination, namely from agricultural, urban and industrial activities, and shellfish generally display a high capacity to bioaccumulate metals, populations may be at risk in terms of toxic metal exposure as a consequence of the harvesting and ingestion of near shore coastal marine organisms. Shellfish is regularly tested for concentrations of metals and other contaminants by legal authorities for commercial purposes, but although health officials use total metal as standards of food safety, only a part of the metal accumulated in shellfish is available to be assimilated and to cause toxic effect. In order to elucidate these issues an investigation on cockles inhabiting the Aveiro estuary was conducted. Element levels in sediments and wild Cerastoderma edule from sampling areas with different levels of contamination were measured; total element burden of cockles was related to accessible fraction for assimilation (TAM); element concentrations in wild C. edule were compared to EFSA (European Food Safe Authorities), USFDA (United States Food and Drug Administration) and FSANZ (Food Standards Australia and New Zealand) maximum levels (MLs); and the amount of cockle flesh needed to be consumed to exceed provisional tolerable weekly intake (PTWI) was determined. The present work showed that although sediment metal and metalloid contamination in Aveiro estuary is low the concentration of elements in C. edule does not reflect the contamination of the sediment. Aluminium (Al) and mercury (Hg) were the less and nickel (Ni), lead (Pb), zinc (Zn) and cadmium (Cd) were the most bioaccumulated metals by cockles. Comparison of MLs from international organisations with the concentration of elements in C. edule showed that arsenic (As) and Pb exceeded standard levels. The ingestion of less than 1 kg for As and 1.5 kg for Pb of cockles would result in exceeding the PTWI threshold (0.015 and 0.025 mg kg⁻¹ week⁻¹ respectively) in any of the areas considered in the study. Cd and Al also appear to be limiting elements for human consumption. Indeed, consumption of more than 3.1 kg and 2.1 kg of whole cockle soft part from one of the study areas during a single week would lead to exceedance of the recommended PTWI value for Cd (0.007 mg kg⁻¹ week⁻¹) and Al (7 mg kg⁻¹ week⁻¹) respectively. The health concerns to humans from cockle consumption from Aveiro estuary are discussed.

Coastal sediments under the influence of multiple organic enrichment sources: An evaluation using carbon and nitrogen stable isotopes

Sediment descriptors (grain size, total volatile solids, redox potential) and stable carbon and nitrogen isotopes were used to trace the origin of organic matter in a coastal area under multiple organic enrichment sources (urban outfall and a major estuary). The sediments fines content and total volatile solids were similar to outfall pre-operation period (1994), but the incorporation of terrestrial organic matter within the sediments located closer to the outfall was diagnosed by depleted (13)C values (-24.2+/-0.38 per thousand) and (15)N values (2.4+/-0.93 per thousand). Data also indicated depleted nitrogen signature at larger distances from the outfall than the carbon signature, due to confounding sediment grain size properties. Analysis in the bivalve Abra alba gave the same results for both isotopes and thus allowed a coherent interpretation of the spatial extent of the organic enrichment, highlighting the importance of bringing a biological element into the environmental assessment.

Horizontal biosedimentary gradients across the Sado estuary, W. Portugal

The topography of the Sado estuary, the second largest of Portugal, comprises the outer estuary inside the entrance channel and the inner estuary, on the inward side of which begins the tidal mudflats. The outer estuary subtidal area covers approximately 70 km2 and presents a series of longitudinal intertidal sandbanks, separating a northern and a southern channel.A benthic survey was undertaken in the outer estuary during June 1986, in which superficial sediments and macrofauna were sampled at 133 locations. The environmental variables measured in the superficial sediments were the temperature, the granulometric structure, the silt, sand and the gravel content, and the total organic matter content. The primary macrofauna biological variables studied were the species composition, abundance and biomass, calculated on wet, dry and ash-free dry weight.The granulometry and the organic content of superficial sediments agreed with the transient and the residual currents velocity field, simulated in a 2-D hydrodynamic model previously elaborated for the outer estuary. The northern channel superficial sediments showed higher silt and total organic matter content, while the model also suggested lower transient and residual velocities, water flow and shear stress in this channel.The distribution patterns of the subtidal macrofauna were separated into two main groups of species, one comprising taxa essentially settled near the estuarine mouth and the other inwards. Biological primary variables also showed consistent patterns, comparable to other Portuguese estuaries. The major subtidal benthic biotopes were obtained through classification analysis and related to the prevailing hydrophysical and sedimentary conditions in the outer estuary.

Estuarine sediment acute toxicity testing with the European amphipod Corophium multisetosum Stock, 1952

This study assessed the use of the European amphipod Corophium multisetosum Stock [Stock, J.H., 1952. Some notes on the taxonomy, the distribution and the ecology of four species of the genus Corophium (Crustacea, Malacostraca). Beaufortia 21, 1-10] in estuarine sediment acute toxicity testing. The sensitivity of adults to the reference toxicant CdCl(2) was determined in water-only 96 h exposures in salinity 2. LC(50) values ranged from 0.33mgCd(2+)L(-1) at 22 degrees C to 0.57mgCd(2+)L(-1) at 15 degrees C. Adult survival was studied in control sediment with water salinity from 0 to 36 and with fine particles content (<63 microm) from 2% to 97% of total sediment, dry weight. Experiments were conducted at 15, 18 and 22 degrees C and the results indicate that the species can be used under the full salinity range although higher mortality was observed at the lower salinity in the higher water temperature, and at the higher salinity in the lower water temperature. The species also tolerated the studied range of sediment fines content and showed the highest sensitivity at intermediate values of fines, especially at the higher temperature, thus advising that tests which have to accommodate sediments with a wide range in fines content should preferably be conducted at 15 degrees C rather than at 22 degrees C. The response in natural sediments was studied in samples collected yearly from 1997 to 2006, at a site located off the Tagus Estuary, western Portugal. A major flood event in winter 2000-2001 induced detectable alterations in sediment baseline descriptors (grain-size, redox potential and total volatile solids), organic contaminants (PAHs, PCBs, DDT metabolites and gamma-HCH) and the macrofauna benthic community. Mortality of the amphipod diminished significantly from the before to the after flood period, in close agreement with diminishing sediment contamination and increasing benthic fauna diversity, in the same time period. C. multisetosum is suitable to conduct acute sediment toxicity tests and presents good potential for the development of a full life-cycle sediment test, due to its amenability to laboratory culture and high survival in the control sediment.

Spatial heterogeneity recognition in estuarine intertidal benthic macrofaunal communities : Influence of sieve mesh-size and sampling depth

Estuarine intertidal soft-bottom macrobenthic infauna of the Tagus estuary was characterised using different mesh size sieves and sediment sampling depth. The study sampled 105 sites using a hand held 0.01 m2 corer. The top layer (0–5 cm) was sieved through nested 1.0 and 0.5 mm meshes whereas the bottom layer (5–20 cm) was through a 1 mm mesh. The total survey took 26 taxa of more than 5800 individuals and a total wet weight biomass of over 650 g. The top layer, using both sieves, gathered 23 taxa (92% of the total), more than 5600 specimens (96%) but less than 8 g of biomass (1%) whereas the 1.0 mm sieve retained 21 taxa (91%), more than 1700 specimens (31%) and almost 7 g of biomass (1%). Abundance was dominated by small annelids, of which Streblospio shrubsolii was 68%, whereas biomass was dominated by molluscs, with the bivalve Scrobicularia plana representing 98%. Multivariate analyses showed an abundance pattern where the top layer data was very similar to that obtained with both layers. The bottom layer data were needed to accurately represent the total biomass pattern. The macrofaunal spatial pattern identified with the 0.5 mm sieve data differed from that identified by the 1.0 mm and was essential to discriminate a faunal assemblage located along the upper part of the shore. It was concluded that in order to characterize the macrofauna community structure, based on the presence/absence of taxa, the top layer and a 1.0 mm sieve would be sufficient. An abundance-based characterization requires the top layer and a 0.5 mm sieve whereas a biomass-based characterization requires data for both layers but it is sufficient to use the 1.0 mm sieve.