Original scientific article UDC 574.587(262.32) Received: 2010-11-01 EVALUATION ON THE ECOLOGICAL STATUS OF THE MACROZOOBENTHIC COMMUNITIES IN THE MARANO AND GRADO LAGOON (NORTHERN ADRIATIC SEA) Nicola BETTOSO, Ida Floriana ALEFFI, Lisa FARESI, Pietro ROSSIN & Giorgio MATTASSI ARPA-FVG Osservatorio Alto Adriático, I-33057 Palmanova (UD), via Cairoli 14, Italy E-mail: nicola.bettoso@arpa.fvg.it Patrizia CRIVELLARO Dipartimento di Scienze della Vita, Universita degli studi di Trieste, I-34100 Trieste, via Valerio 28/A, Italy ABSTRACT The status of the macrozoobenthic community in the Maraño and Grado Lagoon was evaluated, according to the application of the Water Framework Directive for the transitional waters. Benthos samplings were carried out in 2008. At forty-two sampled stations 14,522 organisms from 163 taxa were identified. The number of taxa and diversity indexes decreased from inlets towards the inner bank of the lagoon, as a function of a salinity gradient identified with three water types. Multivariate analysis and analysis of benthic biocenoses revealed the existence of three distinct macrozoobenthic communities related to the closeness and/or the proximity of sea. Dominant species resulted to be typical resident of lagoonal environment, accompanied with opportunistic species able to tolerate large variations of chemical and physical parameters in the transitional environments. M-AMBI index assigned the ecological quality status in relation to biodiversity degree. Key words: macrozoobenthos, Lagoon of Maraño and Grado, ecological status, Water Framework Directive, northern Adriatic Sea VALUTAZIONE DELLO STATO ECOLOGICO DELLE COMUNITA' MACROZOOBENTONICHE NELLA LAGUNA DI MARANO E GRADO (ADRIATICO SETTENTRIONALE) SINTESI Lo stato ecologico delle comunita macrozoobentoniche della Laguna di Marano e Grado e stato esaminato in applicazione della Direttiva Comunitaria sulle acque superficiali per gli ambienti di transizione. Il macrozoobenthos e stato campionato nel 2008 su 42 stazioni e sono stati identificati 14.522 individui per un totale di 163 taxa. Il numero di taxa e gli indici di diversita hanno presentato valori decrescenti dalle bocche lagunari verso le aree interne della laguna, in funzione del gradiente di salinita identificato nei tre tipi idrici. L'analisi multivariata e l'analisi bioce-notica hanno rilevato la presenza di tre distinte comunita bentoniche in relazione alla vicinanza e/o lontananza dal mare. Le specie dominanti sono risultate quelle tipiche degli ambienti lagunari, accompagnate da specie opportuniste capaci di tollerare l'elevata variabilita ambientale degli ambienti di transizione. L'indice M-AMBI ha evidenziato come lo stato di qualita ecologica venga attribuito in funzione del grado di biodiversita. Parole chiave: macrozoobenthos, Laguna di Marano e Grado, stato ecologico, Direttiva Quadro sulle Acque, Adriatico Settentrionale INTRODUCTION Lagoons are classified as transitional waters sited between marine and continental domains. These systems are brackish or hyperhaline water bodies, separated from sea by barrier islands, formed in transgres-sional regime due to the presence of abundant terrigenous supplies and significant coastal transports (Bram-bati, 1988). Until 1950's lagoons, and generally all transitional waters, were classified according to salinity, but the tools employed were quite different. Finally in 1958 the "Venice system" was proposed as a water classification system according to fixed salinity values. The Water Framework Directive (WFD 2000/60/CE) defines transitional waters as "all the surface water bodies in the vicinity of river mouths which are partly saline in character as a result of their proximity to coastal waters but which are substantially influenced by fresh water flows". According to this definition, transitional waters are all ecotones situated between terrestrial, freshwater and marine ecosystems, characterized by high spatial heterogeneity and temporal variability (Basset et al., 2006a). On this basis, transitional waters include fjords, river mouths, deltas, rias, lagoons, coastal ponds and estuaries (McLusky & Elliott, 2007). Transitional waters are heterogeneous and dynamic ecosystems (Gomez et al., 1998; Benedetti-Cecchi et al., 2001) which morphology and hydrology change quickly under the influence of high sedimentation rates, natural coastal dynamics and frequent human activities (Ver et al., 1999; Pastres et al., 2004). These habitats often show high trophic fluxes, broad ranges of chemical and physical parameters with fast biogeochemical cycles (Herbert, 1999; Petihakis et al., 1999; De Wit et al., 2001). In addition, due to their shallow depth and scarce renewal of waters, most of the transitional ecosystems are very vulnerable to eutrophication and chemical pollution (Barnes, 1999), leading to rapid and often unpredictable changes in communities' composition and functioning (Herbert, 1999; Sfriso et al., 2001; Mistri et al., 2002a). The conservation and management of transitional waters requires monitoring activity, integrating chemical and physical evaluation with biological assessment (Gibson et al., 2000; Logan & Furse, 2002). In shallow water systems, such as lagoons, benthic compartment plays a crucial role controlling the main ecological processes; therefore changes in its structure could affect the whole ecosystem (Snelgrove et al., 1997; Weslawski et al., 2004; Tenore et al., 2006). Due to this, it is possible to estimate the effects of different ecological drivers on the ecosystems' functioning, by analyzing modifications of lagoon benthic communities over time (Pranovi et al., 2008). The Marano and Grado Lagoon is a part of the lagoon system of the northern Adriatic Sea, stretching between the mouths of the Po and Isonzo rivers (Bram- bati, 1988). The lagoon, which is located among the Isonzo river to the East and the Tagliamento river to the West, has a total surface area of 160 km2, and extends parallel to the coastline along 32 km (Falace et al., 2009). Aristocle Vatova has provided most of the main studies and notions dealing with structure of benthic communities in the northern Adriatic lagoons. Since 1930 he studied hydrology, benthic flora and fauna in the Venice Lagoon (Vatova, 1940, 1949), subsequently benthic fauna and productivity of the Marano and Grado Lagoon (Vatova, 1964a, 1964b, 1965). Vatova (1964a) described the general features of the Marano and Grado lagoons, focusing on differences between the two basins. In particular the author measured a lower mean salinity in the Marano Lagoon (21%o) than the Grado Lagoon (26%o) and this difference allowed distinguishing the basins and the related distribution of benthic communities. Benthic fauna of the Marano was in fact poorer than that of Grado, both in term of species and abundance, due to higher freshwater inputs. On the other hand, it was more productive in term of biomass. The most recent and exhaustive study on macrozoo-benthos in the Marano and Grado Lagoon was carried out during a three-year study, from 1993 to 1995 (Orel et al., 2001; Zamboni, 2008). Authors substantially confirmed the observations pointed out by Vatova (1964a): a decreasing gradient of biodiversity was observed moving from the Grado to the Marano, as well as from inlets to inner areas of the lagoon. Orel et al. (2001) and Zamboni (2008) identified the zonation of benthos on the basis of confinement degree in the paralic environments as proposed by Guelorget & Perthuisot (1983), who defined paralic environments the aquatic ecosystems which have, or had, relation with the sea. In 1986 the Manila clam (Tapes philippinarum) was introduced in the Marano Lagoon for aquaculture purposes. In the time its irrational harvesting with mechanical dredges out of farming areas impacted the benthic community (Orel et al., 2002, 2005). Since 2006 the uncontrolled use of dredges was stopped and manual harvesting is permitted beyond the farming areas. The aim of the paper was to evaluate the ecological status of the macrozoobenthic community in the Marano and Grado Lagoon, as required by the application in the WFD. MATERIAL AND METHODS The Marano and Grado Lagoon is defined as a coastal microtidal lagoon with large dimensions (Italian Ministry of Environment Decree n.131/08) and the related water types were established as a function of salinity values. In detail, we can discriminate between the mesohaline lagoon (5-20 psu), polyhaline lagoon (2030 psu) and euhaline lagoon (30-40 psu). Forty-two Rivera Q Sampling dations Water body I y peí Mesoha line f I Polyhaline I I Eu ha line I I Heavily modified Fig. 1: Study area and sampling stations. SI. 1: Obravnavano območje in vzorčevalne postaje. sampling sites were selected both on the basis of water types, surface and potential gradient of confinement from sea inlets to inner areas (Fig. 1 ). Benthic samples were collected with a 0.047 m2 van Veen grab in May 2008. At each station four grabs were taken. The sediment was sieved on a 1 mm mesh and fixed in 4% buffered formaldehyde solution stained with Bengal Rose, and then the fauna was separated and identified to the lowest possible taxonomical level. Uni- and multivariate techniques were employed to analyze the communities' structure including: abundance, number of taxa, diversity indexes (ShannonWiener diversity index (H') on log2 basis (Shannon & Weaver, 1949), Margalef's index (d) (Margalef, 1958) and Pielou's evenness index (J) (Pielou, 1966). The Bray-Curtis similarity coefficient was calculated on square-root transformed data, using complete linkage; subsequently, one-way ANOSIM, .-dominance curves and SIMPER analysis were applied to evaluate similarity and/or differences among groups (PRIMER software package developed at the Plymouth Marine Laboratory). Bionomic percentage affinity (A%) was calculated by considering characteristic species according to Pérès & Picard (1964). The correction coefficient C was first calculated as a percentage of characteristic species of bio- cenosis j respect to the ones of other biocenosis. Then, the absolute affinity of each station was calculated as: A¡= n¡ (100-Cj) where n¡ is the number of characteristic species of biocenosis j in the considered station. Finally, using a simple proportion, this parameter was expressed as percentage affinity (A%). A% was calculated for each biocenosis found in the lagoon: Euryhaline and Eurythermal Lagoon biocenosis (LEE French acronym for biocoenose Lagunaire Eury-haline et Eurytherme), fine well-sorted sand biocenosis (SFBC biocoenose des Sables Fins Bien Calibres), superficial muddy sand in sheltered areas biocenosis (SVMC biocoenose des Sables Vaseux Superficiels en Mode Calme), coastal terrigenous muds biocenosis (VTC biocoenose des Vases Terrigenes Cotieres), Posidonia oceanica meadow biocenosis (HP biocoenose de l'Herbier de Posidonies), fine superficial sand biocenosis (SFS biocoenose des Sables Fins Superficiels) and coastal detriti-cal bottoms biocenosis (DC biocoenose des fonds Detritiques Cotiers). The Bray-Curtis similarity coefficient was calculated on not transformed A% data, using complete linkage. Furthermore RELATE procedure was used to compare clusters derived from abundance and A% data. Tab. 1: Average values of number of taxa, abundance, H', d, - and summary of Kruskal-Wallis one-way analysis of variance applied to the water types. Tab. 1: Povprečne vrednosti števila taksonov, številčnosti, H', d, - in povzetek Kruskal-Wallisove enosmerne analize variance na tipih voda. No. taxa Abundance (ind./m2) H' d J Euhaline 38±14 1,775±1,773 3.79±0.68 6.46±2.00 0.74±0.10 Polyhaline 26±8 1,937±1,698 3.01 ±0.81 4.37±1.19 0.65±0.15 Mesohaline 13±4 1,711±892 2.13±0.50 2.06±0.70 0.59±0.09 Kruskal-Wallis (H) 18.49 0.026 16 20.2 5.23 p-value <0.0001 <0.99 <0.0003 <0.0001 <0.073 df 2 2 2 2 2 AMBI and Biotic Index (Bl) were applied (Borja et a/., 2000) using the AMBI program (AZTI Marine Biotic Index) (www.azti.es). These indexes are based on the classification of the benthic species in five (I-V) ecological groups (EG), according to their tolerance to pollution (from EG-I = species very sensitive to organic enrichment, intolerant to pollution, EG-II = species indifferent to enrichment, EG-III = species tolerant to enrichment, slightly unbalanced environments, EG-IV = second-order opportunistic species, slight to pronounced unbalanced environments, to EG-V = first-order opportunistic species, pronounced unbalanced environment), then applying an algorithm to calculate the AMBI on a scale of increasing pollution (from 1 to 6) and obtaining the corresponding BI (from 0-1 = unpolluted sites, 2 = slightly polluted, 3 = moderately polluted, 4-5 = moderately to heavily polluted, 6 = heavily polluted to 7 = extremely polluted, azoic state). M-AMBI (Multivariate AMBI) was calculated to assess the ecological quality status (EcoQS): High, Good, Moderate, Poor and Bad according to WFD. This index includes the species richness, Shannon-Wiener diversity and AMBI at the very same time (Muxika et a/., 2007). EcoQS was evaluated on the basis of the reference conditions proposed for Italian transitional waters (ISPRA, 2010). RESULTS 14,522 organisms from 163 taxa (142 species determined) were identified. Polychaetes were by far the dominant group with 72 species followed by the molluscs (36 species), crustaceans (25 species), echinoderms (6 species) and "other". This latter usually represents scarce groups such as ascidians, anthozoans, sipuncu-lids, nemertines, phoronids, turbellarians and larvae of insects. Taking into consideration the three water types, 147 taxa were recorded in the euhaline water type (16 sampling stations) for a total of 5,339 individuals; 106 taxa were collected in the polyhaline water type (20 sampling stations), for a total of 7,283 individuals and finally 36 taxa in the mesohaline water type (6 sampling stations), for a total of 1,930 individuals. The percentage abundance of groups was quite proportional in each water type, but the disappearance of echinoderms in the mesohaline lagoon was notable (Fig. 2). Table 1 shows mean values in each water type of the number of taxa, abundance, H', d and J. A clear decreasing gradient from euhaline lagoon to mesohaline basin was significative for taxa, H' and d (Tab. 1); mean abundance did not show any gradient from inlet to inner areas, whereas mean J was decreasing but not in a significative manner. Fig. 2: Percentage of taxa for different animal phyla detected in euhaline, polyhaline and mesohaline water types. Sl. 2: Delež taksonov za različna živalska debla v evhalinem, polihalinem in mezohalinem tipu vode. The dendrogram resulting from the Bray-Curtis similarity matrix showed three different groups tested with one-way ANOSIM (R=0.578; p<0.001): stations close to inlets (group 1), stations among inlets and the inner bank (group 2) and stations closed to the inner bank (group 3) (Fig. 3). Fig. 3: Dendrogram obtained by taxa abundance values. SI. 3: Dendrogram številčnosti taksonov. Tab. 2: Cumulative percentage of dominant taxa in each group revealed by SIMPER analysis. Tab. 2: Kumulativni odstotek dominantnih taksonov v vsaki skupini, ugotovljen s SIMPER analizo. Group Taxa Cum. % Paralic group Abra segmentum 34.7 Hediste diversicolor 55.05 Streblospio shrubsolii 66.94 Mixed group Chaetozone sp. 17.75 Abra segmentum 34.76 Heteromastus filiformis 44.91 Streblospio shrubsolii 53.34 Oligochaeta indet. 61.48 Marine group Minuspio cirrifera 12.02 Mediomastus capensis 21.97 Heteromastus filiformis 30.16 Abra segmentum 37.55 Pseudoleiocapitella fauveli 44.59 Capitella capitata 51.12 Oligochaeta indet. 56.84 Myriochele oculata 62.1 Trend of k-dominance curves identified the community structure in each of the identified groups: group 1, or marine group, with the highest number of species and a quite regular and homogeneous trend; group 3, or paralic group, with higher slope and the lowest number of species; group 2, or mixed group, with an intermediate shape (Fig. 4). SIMPER analysis identified the mostly involved species in the three groups (Tab. 2). The paralic group was mainly represented by the bivalve Abra segmentum, the nereid polychaet Hediste diversicolor and the spionid Streblospio shrubsolii. The mixed group was character- ized by the abundance of the capitellid Heteromastus fi-liformis, the cirratulid Chaetozone sp. and oligochaets, in addition to A. segmentum and S. shrubsolii. The Marine group, in addition to A. segmentum, H. filiformis and oligochaets, recorded the abundance of the capitel-lids Mediomastus capensis, Pseudoleiocapitella fauveli, Capitella capitata, the spionid Minuspio cirrifera and the owenid Myriochele oculata. Overall, 26 exclusive and preferential species for 7 biocenoses were found (Tab. 3). The dendrogram obtained from the Bray-Curtis similarity matrix applied on A% data, highlighted three groups of stations on the ba- sis of the different A% values for Eurythermal and Eury-haline biocoenosis (LEE) (Fig. 5): the Paralic group having A°/olee >65%, the Marine group with A%LEE <30% and Mixed group 30%< A%LEE <65%. The RELATE procedure revealed a similarity of the groups identified by two dendrograms (Rho=0.546; p<0.0001). Tab. 3: Characteristic species and biocenoses detected in the study area: Euryhaline and Eurythermal Lagoon biocenosis (LEE), fine well-sorted sand biocenosis (SFBC), superficial muddy sand in sheltered areas biocenosis (SVMC), coastal terrigenous muds biocenosis (VTC), Posidonia oceanica meadow biocenosis (HP), coastal detritical bottoms biocenosis (DC) and fine superficial sand biocenosis (SFS). Tab. 3: Značilne vrste in biocenonoze v obravnavanem območju: biocenoza evrihaline in evritermne lagune (LEE), biocenoza na finem sortiranem pesku (SFBC), biocenoza zamuljenih peskov v zaščitenih predelih (SVMC), biocenoza obalnega terigenega mulja (VTC), biocenoza podvodnih travnikov Posidonia oceanica (HP), biocenoza obalnega detritičnega dna (DC) in biocenoza finega površinskega peska (SFS). The EcoQS assigned with M-AMBI, was Good for 44% of euhaline stations, Moderate for 38%, Poor for 12% (st. 76 and 100) and Bad for 6% (st. 99) (Fig. 6). In polyhaline type the stations with High status corresponded to 20%, Good to 50%, Moderate to 25% and Poor to 5% (st. 86) (Fig. 6). In the mesohaline type one station (st. 43) was Good, three stations were assessed as Moderate (50%) and two were Poor (st. 13 and 17) (Fig. 6). The summarizing Table 4 reports M-AMBI as EcoQS, AMBI as disturbance classification and the groups (Marine, Mixed, Paralic) assigned with clusters. The mean number of taxa and average Shannon-Wiener index (H') were calculated for each EcoQS of the water types. Within the euhaline type (salinity 30-40 psu) all stations with Good ecological quality status possessed Marine characteristics, with 50 identified taxa and H' >4. Stations with Moderate EcoQS possessed normally Mixed characteristics, on average 29 taxa and mean H'=3.5. Stations having Poor EcoQS possessed 33 taxa and H'=2.9: st. 100 had Marine characteristics but a Poor status was assigned probably because of high dominance of oligochaetes, indicating strongly unbalanced conditions. Bad EcoQS was assigned to st. 99, which was very poor in term of taxa. In the polyhaline type (salinity 20-30 psu) only two stations have Marine characteristics (st. 25 and 35), but normally stations showed Mixed or sometimes Paralic conditions. Stations with High EcoQS possessed on average 38 taxa and H'=4. Stations with Good EcoQS had 24 taxa and H'>3. In mesohaline type (salinity 5-20 psu), stations have Paralic conditions, except for st. 43 which is Mixed-Paralic with Good EcoQS (17 taxa, H'=2.5). Disturbance classification detected with AMBI varied from slightly disturbed to moderately disturbed and it seems not linked to water type and cluster groups. DISCUSSION The aim of this study was to characterize the macro-zoobenthos living in the Marano and Grado Lagoon, 15 years after the studies carried out in 1993-1995 (Drioli, 1995/1996; Zamboni, 1995/1996; Orel et al., 2001; Zamboni, 2008), and to provide a preliminary application of the WFD 2000/60/CE in this lagoon. Previously, several authors classified lagoons and coastal ponds as a function of salinity values (Redeke, 1922, 1933; Brunelli, 1933; D'Ancona, 1959). The salinity is a key parameter affecting the biological organization and the expression of biodiversity in Italian lagoons (Basset et al., 2006b) and Greek brackish environments (Reizopoulou & Nicolaidou, 2004). Three water types were identified and analyzed in Marano and Grado according to the actual Italian legislations. Characteristic species Biocenosis Gibbula adriatica LEE Cerastoderma glaucum Abra segmentum Tapes philippinarum Hediste diversicolor Streblospio shrubsolii Carcinus mediterraneus Nephtys hombergi SFBC Owenia fusiformis Prionospio caspersi Diogenes pugilator Tellimya ferruginosa Thracia papyracea Euclymene oerstedi Upogebia pusilla SVMC Loripes lacteus Tapes decussates Paphia aurea Petaloproctus terricolus Sternaspis scutata VTC Ampharete acutifrons Laonice cirrata Venus verrucosa HP Euclymene lumbricoides Abra prismatica DC Glycera tridactyla SFS Tab. 4: Summarizing table. Tab. 4: Pregledna tabela. sampling station water type EcoQS No. taxa H' groups cluster groups cluster AMBI (M-AMBI) (mean values) (mean values) A% abundance disturbance classification 46 EU GOOD 50 4.4 Marine Marine Slightly 56 EU Marine Marine Slightly 59 EU Marine Marine Slightly 73 EU Marine Marine Moderately 78 EU Mixed Marine Moderately 80 EU Marine Marine Moderately 84 EU Marine Mixed Slightly 29 EU MODERATE 29 3.5 Mixed Mixed Slightly 38 EU Mixed Mixed Moderately 52 EU Marine Paralic Slightly 89 EU Mixed Mixed Moderately 92 EU Mixed Mixed Moderately 97 EU Mixed Marine Slightly 76 EU POOR 33 2.9 Mixed Mixed Moderately 100 EU Marine Marine Moderately 99 EU BAD 14 3 Mixed Mixed Moderately 2 POLY HIGH 38 4 Paralic Marine Moderately 23 POLY Paralic Paralic Slightly 31 POLY Mixed Mixed Slightly 53 POLY Paralic Mixed Slightly 5 POLY GOOD 24 3.2 Mixed Mixed Slightly 19 POLY Mixed Mixed Moderately 20 POLY Mixed Mixed Moderately 26 POLY Paralic Paralic Moderately 34 POLY Mixed Paralic Slightly 45 POLY Mixed Mixed Moderately 60 POLY Paralic Paralic Slightly 62 POLY Mixed Paralic Slightly 65 POLY Mixed Mixed Moderately 71 POLY Mixed Mixed Moderately 6 POLY MODERATE 22 2.2 Paralic Paralic Moderately 25 POLY Marine Marine Slightly 35 POLY Marine Marine Moderately 82 POLY Paralic Mixed Moderately 91 POLY Mixed Mixed Moderately 86 POLY POOR 10 1.6 Paralic Paralic Slightly 43 MESO GOOD 17 2.5 Mixed Paralic Slightly 10 MESO MODERATE 14 2.4 Paralic Paralic Slightly 22 MESO Paralic Paralic Slightly 44 MESO Paralic Paralic Moderately 13 MESO POOR 9 1.6 Paralic Paralic Slightly 17 MESO Paralic Paralic Slightly Fig. 4: K-dominance curves relative to three different groups detected by dendrogram. SI. 4: Krivulje K-dominance za tri različne skupine, ugotovljene z dendrogramom. The Marano Lagoon is embedded in the inland, coupled to a less marine water exchange and very abundant freshwater inputs from Stella, Corno and Aussa rivers (Fig. 1); on the contrary the Grado Lagoon is characterized by scarce freshwater inputs and a wider exchange of marine water. The subdivision of the whole basin into three water types reflects quite well the previous observations of Vatova (1964a, 1964b, 1965) and more recently those of Orel et al. (2001) and Zamboni (2008). During 2008 taxa richness, diversity and evenness indices showed high values particularly in nearby inlets. In these areas Shannon-Wiener index (H') was >4, which can be considered a very high value for soft bottom macrozoobenthic communities (Gray, 2000). Therefore high indices values and relevant number of species recorded in 2008 (163 taxa) compared to the 90's, (85 taxa in the three years; Zamboni, 2008), could indicate the increasing richness of taxa in the Marano and Grado Lagoon during the last 15 years. The number of taxa decreased moving from the inlets towards the inner bank of the lagoon, as described also in previous studies. On the contrary, abundance did not show any distribution gradient. On the whole, the partition of benthos into three salinity bands was confirmed by a decreasing diversity from the inlets towards the inner bank. Due to this, a monitoring program based on water types is fully supported by observations carried out in the first studies (Vatova, 1964a, 1965; Orel et al., 2001; Zamboni, 2008) and the 2008 survey. Guelorgèt & Perthuisot (1982, 1983, 1992) referred on the existence of peculiar euryhaline species. This feature was previously emphasized by Pérès & Picard (1964), who defined the Euryhaline and Eurythermal Lagoon biocenosis. This biocenosis is typical of unstable environments where higher or lower salinity and/or wider salinity and temperature variations occur during the year. This peculiarity is mainly due to river floods, rainfalls and high summer rate of evaporation. Moreover, living communities are able to recover rather quickly their original structures when environmental disturbances and/or dystrophic crisis occur. Due to this great resilience, rather than stability of the communities, paralic environments prevail. The previously mentioned peculiar characteristics of the communities led Guelorget & Perthuisot (1982, 1983, 1992) to divide the paralic domain into two subsets. The first one as near paralic, close to the inlets and with chemical-physical parameters similar to sea, whereas the second one as far paralic, farther from sea with chemical-physical parameters deeply different to marine domain. As previously discussed, the salinity is considered a key parameter determining a transition gradient from a typical marine community to a freshwater one. In fact, the fresh/marine waters exchange and circulation play an important role in the dynamics of a lagoon. In this way, hydrology is the fundamental factor conditioning any lagoon environment. In the same context Guelorget & Perthuisot (1992) defined the confinement as the time of renewal of Fig. 5: Dendrogram obtained by percentage affinity values. SI. 5: Dendrogram, narejen na podlagi afinitetnih vrednosti (v %). marine elements in relation to the extension of paralic domain. However, this definition should be considered on a larger scale since it is dependent on many other parameters such as tidal range, width inlets, freshwaters inputs, wind regime, depth, trophic conditions, transparency, oxygen content and so on. Multivariate analysis in 2008 defined three different areas as a function of closeness to inlets and freshwaters inputs. The group of stations close to inlets could be defined as a lagoon community having marine characteristics; stations nearby the inner bank as a strictly paralic community; finally stations showing a combination of marine and paralic properties as a mixed community. This kind of partition was confirmed and validated by ANOSIM analysis; moreover k-dominance curves focused a basic community structure in each group. In the same way SIMPER analysis punctually identified the mostly involved species of the mentioned groups. Three characteristic species of paralic environments and LEE biocenosis represent at least 60% of relative abundance in the Paralic group and dominate in inner zones where marine species cannot survive: Abra seg-mentum, Hediste diversicolor and Streblospio shrubsolii. A. segmentum is typically euryhaline, frequent in oligo-hyperhaline waters, tolerates a wide range of salinities, from 3 to 41 psu (Marazanof, 1969; Kevrekidis, 2004). It is common and frequently abundant in the Mediterranean coastal lagoons, where it plays a dominant role, both in terms of number and biomass, in the infauna of these habitats and represents an important food for fish (Kevrekidis & Kasapis, 2009). H. diversicolor preferentially lives in muddy sediments. It is a typical inhabitant of European brackish water habitats and shows a high level of tolerance towards different types and concentrations of contaminants (Volpi et al., 1999). H. diversi-color tolerates a wide salinity range, preferring, in any case, areas characterized by low salinity values (Guer-zoni & Tagliapietra, 2006) and it is largely distributed in Mediterranean lagoons (BazaTri et al., 2003; Nicolaidou et al., 2005). S. shrubsolii is a typical lagoon species widely distributed also in other Mediterranean lagoons (Mistri et al., 2002b; Rossi & Lardicci, 2002; BazaTri et al., 2003; Dauer et al., 2003). Furthermore, Mixed and Marine groups were characterized by the dominance of polychaetes belonging to Capitellidae and Spionidae families represented by small size species capable to settle in habitats having a strong variability, such as transitional environments (Holte & Oug, 1996; Mistri et al., 2002b) and adapted to colonize habitats with organic enrichment (Mistri et al., 2001; Thouzeau et al., 2007). Oligochaets, indicating desalinized water (Nicolaidou et al., 2005), were abundant, too. In addition, the owenid Myriochele oculata, a sandy species especially diffused nearby inlets, contributed to define the Marine group. The bionomic analysis (Péres & Picard, 1964) revealed the existence of seven biocenoses. The main were LEE and SFBC, this latter is typical of marine domain and largely diffused in Mediterranean Sea as a soft bottom infralittoral biocenosis. The second dendrogram obtained by A% values was comparable to abundance cluster as revealed by RELATE procedure. In this way stations having Marine affinities are mostly represented by SFBC, whereas LEE is meanly <30%. LEE affinity in Paralic stations exceeds 65%, and LEE affinity in Mixed group ranges between 30 and 65%. The Marine, Mixed and Paralic groups could correspond to zones II, III and IV-V respectively, as defined by Guelorget & Perthuisot's benthic zonation (Guelorget & Perthuisot, 1982, 1983, 1992). In fact, Marine group resembles the zone II, as a corresponding entrance into lagoon domain with the presence of most tolerant marine species belonging to SFBC and Fine Superficial Sands (SFS) biocenosis. Mixed group was similar to zone III with a remarkable scarcity or even disappearance of echinoderms. Vatova (1964a, 1965) noticed that the holothurioid Trachythyone elongata is the only capable to withstand sudden salinity changes and, in fact, only 7 specimens of 4 echinoderms species were found in the polyhaline water types (T. elongata, Asterina gibbosa, Amphiura chiajei and Amphipholis squamata). Finally, Paralic group resembled zone IV, because of the total disappearance of most tolerant marine species, such as echinoderms, and an absolute dominance of strictly paralic species such as A. segmentum, Cerastoderma glaucum and H. diversicolor. In some limited areas Paralic group resembled also zone V because of the appearance of chironomids larvae. No specimens of the characteristic lagoon bivalve Scrobicularia plana were found in 2008. This species already recorded in the Marano and Grado Lagoon by Vatova (1964a, 1964b, 1965) and considered as particularly sensitive to river floods, was scarcely present in the 70's because of pollution phenomena (Battaglia et al., 1972) and now it results to be absent since the 90's (Zamboni, 2008). Macrozoobenthos in 2008 embraced a distribution comparable to the three years study 1993-1995 (Orel et al., 2001; Zamboni, 2008). In the lagoon the salinity constitutes a sort of barrier to strictly marine species, leading to the selection of more tolerant organisms. In the inner part of the lagoon there is a clear dominance of LEE species accompanied by strongly opportunistic annelids. The final goal was the application of indices proposed by WFD, in order to define the ecological quality status in each sampling station. M-AMBI showed a significant Good-Moderate quality status in euhaline stations. However, some sites located within heavily modified water bodies showed a Poor or Bad status. In polyhaline type 70% of stations showed Good to High EcoQS. Mesohaline type, the inner part of the Marano Lagoon, showed almost a Moderate status, due to natural selectivity of paralic habitats, characterized by low biodiversity. As suggested by Dauvin et al. (2009), the main issue in the application of indexes for the classification is that in order to determine anthropogenic stress, Fig. 6: EcoQS of euhaline, polyhaline and mesohaline stations according to M-AMBI index. Sl. 6: Ekološko stanje kakovosti (EcoQS) za evhaline, polihaline in mezohaline postaje na podlagi izračuna indeksa M-AMBI. they account to relative abundances of stress-tolerant species. However, these latter may also be tolerant to natural stressors. Moreover, due to the high variability of both physical and chemical parameters, transitional environments are generally characterized by low benthic diversity if compared to the marine environment with several opportunistic and tolerant organisms, withstanding these conditions. AMBI provides ecological group assignments based on expert opinion for a lot of taxa. Moreover the ecological behaviour of several species is quite different when they have to adapt to la-goonal conditions (Cognetti & Maltagliati, 2008) or to different geographical areas. The reference conditions for the final EcoQS are established by the legislation. Actually, in Italy, a draft document proposed by Italian Environmental Ministry provides the same reference condition for oligo-, meso- and polyhaline waters. As aforementioned the Marano and Grado Lagoon shows a quite different benthic community in the mesohaline waters compared to polyhaline waters. Hence the application of a single reference condition for these different types causes to classify mesohaline waters with Moderate or Poor quality. In this situation, in order to achieve at least the Good EcoQS in the whole Marano and Grado Lagoon, macrozoobenthic community should reach a number of species comparable to a typical soft bottom community of marine environments, thus loosing the characteristics and ecological functions of paralic environments. ACKNOWLEDGEMENTS Authors are grateful to Dr. Alessandro Acquavita and two anonymous referees for the critical evaluation of the manuscript. We dedicate this paper to the memory of Ziga Dobrajc and Samo Alajbegovic, who tragically passed away last summer. OCENA EKOLOŠKEGA STATUSA ZDRUŽB VODNIH NEVRETENČARJEV V MARANSKI IN GRADEŠKI LAGUNI (SEVERNI JADRAN) Nicola BETTOSO, Ida Floriana ALEFFI, Lisa FARESI, Pietro ROSSIN & Giorgio MATTASSI ARPA-FVG Osservatorio Alto Adriatico, I-33057 Palmanova (UD), via Cairoli 14, Italy E-mail: nicola.bettoso@arpa.fvg.it Patrizia CRIVELLARO Dipartimento di Scienze della Vita, Universita degli studi di Trieste, I-34100 Trieste, via Valerio 28/A, Italy POVZETEK Ocenjenjen je bil status združb vodnih nevretenčarjev v Maranski in Gradeški laguni, v skladu z aplikacijo Okvirne direktive o vodah za somornice. Vzorčenja bentosa so bila izvedena leta 2008. Na 42 vzorčevalnih postajah je bilo identificiranih 14.522 organizmov, pripadajočih 163 taksonom. Število taksonov in indeksi pestrosti so se zman-ševali od mesta dotokov proti notranjemu bregu lagune, in sicer kot funkcija gradienta slanosti treh tipov vode. Mul-tivariatna analiza in analiza bentoških biocenoz sta razkrili obstoj treh združb vodnih nevretenčarjev, vezanih na bližino/oddaljenost od morja. Dominantne vrste so tipični prebivalci lagunskega okolja, spremljajo pa jih oportunis-tične vrste, ki lahko tolerirajo velike variacije kemičnih in fizikalnih parametrov v somornicah. Indeks M-AMBI opredeljuje ekološko stanje glede na biotsko raznolikost. Ključne besede: vodni nevretenčarji, Maranska in Gradeška laguna, ekološki status, Okvirna direktiva o vodah, severni Jadran REFERENCES Barnes, R. S. K. (1999): What determines the distribution of coastal hydrobiid mudsnails within north-western Europe. Mar. Ecol. P.S.Z.N.I., 20, 97-110. Basset, A., L. Sabetta, A. Fonnesu, D. Mouillot, T. Do Chi, P. Viaroli, G. Giordani, S. Reizopoulou, M. Abbiati & G. C. Carrada (2006a): Typology in Mediterranean transitional waters: new challenger and perspective. Aquat. Conserv.: Mar. Freshw. Ecosyst., 16, 441-455. Basset, A., N. Galuppo & L. 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