Effect of multiple sources of disturbance on rocky benthic assemblages in some localities of Salento, Apulia

The first part of my work consisted in samplings conduced in nine different localities of the salento peninsula and Apulia (Italy): Costa Merlata (BR), Punta Penne (BR), Santa Cesarea terme (LE), Santa Caterina (LE), Torre Inserraglio (LE), Torre Guaceto (BR), Porto Cesareo (LE), Otranto (LE), Isole Tremiti (FG). I collected data of species percentage covering from the infralittoral rocky zone, using squares of 50x50 cm. We considered 3 sites for location and 10 replicates for each site, which has been taken randomly. Then I took other data about the same places, collected in some years, and I combined them together, to do a spatial analysis. So I started from a data set of 1896 samples but I decided not to consider time as a factor because I have reason to think that in this period of time anthropogenic stressors and their effects (if present), didn’t change considerably. The response variable I’ve analysed is the covering percentage of an amount of 243 species (subsequently merged into 32 functional groups), including seaweeds, invertebrates, sediment and rock. 2 After the sampling, I have been spent a period of two months at the Hopkins Marine Station of Stanford University, in Monterey (California,USA), at Fiorenza Micheli's laboratory. I've been carried out statistical analysis on my data set, using the software PRIMER 6. My explorative analysis starts with a nMDS in PRIMER 6, considering the original data matrix without, for the moment, the effect of stressors. What comes out is a good separation between localities and it confirms the result of ANOSIM analysis conduced on the original data matrix. What is possible to ensure is that there is not a separation led by a geographic pattern, but there should be something else that leads the differences. Is clear the presence of at least three groups: one composed by Porto cesareo, Torre Guaceto and Isole tremiti (the only marine protected areas considered in this work); another one by Otranto, and the last one by the rest of little, impacted localities. Inside the localities that include MPA(Marine Protected Areas), is also possible to observe a sort of grouping between protected and controlled areas. What comes out from SIMPER analysis is that the most of the species involved in leading differences between populations are not rare species, like: Cystoseira spp., Mytilus sp. and ECR. Moreover I assigned discrete values (0,1,2) of each stressor to all the sites I considered, in relation to the intensity with which the anthropogenic factor affect the localities. 3 Then I tried to estabilish if there were some significant interactions between stressors: by using Spearman rank correlation and Spearman tables of significance, and taking into account 17 grades of freedom, the outcome shows some significant stressors interactions. Then I built a nMDS considering the stressors as response variable. The result was positive: localities are well separeted by stressors. Consequently I related the matrix with 'localities and species' with the 'localities and stressors' one. Stressors combination explains with a good significance level the variability inside my populations. I tried with all the possible data transformations (none, square root, fourth root, log (X+1), P/A), but the fourth root seemed to be the best one, with the highest level of significativity, meaning that also rare species can influence the result. The challenge will be to characterize better which kind of stressors (including also natural ones), act on the ecosystem; and give them a quantitative and more accurate values, trying to understand how they interact (in an additive or non-additive way).

Effects of Paramuricea clavata (Risso, 1826) (Anthozoa: Plexauridae) forests on settlement of benthic species: an experimental approach

Rationale: Coralligenous habitat is considered the second most important subtidal “hot spot” of species diversity in the Mediterranean Sea after the Posidonia oceanica meadows. It can be defined as a typical Mediterranean biogenic hard bottom, mainly produced by the accumulation of calcareous encrusting algae that, together with other builder organisms, form a multidimensional framework with a high micro-spatial variability. The development of this habitat depends on physical factors (i.e. light, hydrodynamism, nutrients, etc.), but also biologic interactions can play a relevant role in structuring the benthic assemblages. This great environmental heterogeneity allows several different assemblages to coexist in a reduced space. One of the most beautiful is that characterised by the Mediterranean gorgonian Paramuricea clavata (Risso, 1826) that can contribute to above 40% of total biomass of the community and brings significant structural complexity into the coralligenous habitat. In sites moderately exposed to waves and currents, P. clavata can form high-density populations (up to 60 colonies m-2) between 20 – 70 m in depth. Being a suspension feeder, where it forms dense populations, P. clavata plays a significant role in transferring energy from planktonic to benthic system. The effects of the branched colonies of P. clavata could be comparable to those of the forests on land. They can affect the micro scale hydrodynamism and light, promoting or inhibiting the growth of other species. Unfortunately, gorgonians are threatened by several anthropogenic disturbance factors (i.e. fishing, pollution, tourism) and by climatic anomalies, linked to the global changes, that are responsible of thermal stress, development of mucilage and enhanced pathogens activity, leading to mass mortality events in last decades. Till now, the possible effects of gorgonian forest loss are largely unknown. Our goal was to analyse the ecological role of these sea fan forests on the coralligenous benthic assemblages. Experimental setup and main results: The influence of P. clavata in the settlement and recruitment of epibenthic organisms was analysed by a field experiment carried out in two randomly selected places: Tavolara island and Portofino promontory. The experiment consisted in recreate the presence and absence of the gorgonian forest on recruitment panels, arranged in four plots per type (forested and non-forested), interspersed each other, and deployed at the same depth. On every forested panel 3 gorgonian colonies about 20 cm height were grafted with the use of Eppendorf tubes and epoxy resin bicomponent simulating a density of 190 sea fans per m-2. This density corresponds to a mean biomass of 825 g DW m-2,3 which is of the same order of magnitude of the natural high-density populations. After about 4 months, the panels were collected and analysed in laboratory in order to estimate the percent cover of all the species that have colonized the substrata. The gorgonian forest effects were tested by multivariate and univariate permutational analyses of the variance (PERMANOVA). Recruited assemblages largely differed between the two study sites, probably due to different environmental conditions including water quality and turbidity. On overall, the presence of P. clavata reduced the settlement and recruitment of several algae: the shadow caused by the gorgonian might reduce light availability and therefore their growth. This effect might be greater in places where the waters are on average more clear, since at Portofino it is less visible and could be masked by the high turbidity of the water. The same pattern was registered for forams, more abundant outside gorgonian forest, probably linked with algal distribution, shadowing effect or alimentary competition. The last one hypothesis could be valid also for serpulids polychaetes that growth mainly on non-forested panels. An opposite trend, was showed by a species of bryozoan and by an hydroid that is facilitated by the presence of P. clavata, probably because it attenuates irradiance level and hydrodynamism. Species diversity was significantly reduced by the presence of P. clavata forests at both sites. This seems in contrast with what we expected, but the result may be influenced by the large algal component on non-forested panels. The analysis confirmed the presence of differences in the species diversity among plots and between sites respectively due to natural high variability of the coralligenous system and to different local environment conditions. The reduction of species diversity due to the presence of gorgonians appeared related to a worst evenness rather than to less species richness. With our experiment it is demonstrated that the presence of P. clavata forests can significantly alter local coralligenous assemblages patterns, promoting or inhibiting the recruitment of some species, modifying trophic relationships and adding heterogeneity and complexity to the habitat. Moreover, P. clavata could have a stabilising effect on the coralligenous assemblages.

Natural patterns and anthropogenic disturbance in north Adriatic marine benthic assemblages: descriptive and methodological studies

Marine soft bottom systems show a high variability across multiple spatial and temporal scales. Both natural and anthropogenic sources of disturbance act together in affecting benthic sedimentary characteristics and species distribution. The description of such spatial variability is required to understand the ecological processes behind them. However, in order to have a better estimate of spatial patterns, methods that take into account the complexity of the sedimentary system are required. This PhD thesis aims to give a significant contribution both in improving the methodological approaches to the study of biological variability in soft bottom habitats and in increasing the knowledge of the effect that different process (both natural and anthropogenic) could have on the benthic communities of a large area in the North Adriatic Sea. Beta diversity is a measure of the variability in species composition, and Whittaker’s index has become the most widely used measure of beta-diversity. However, application of the Whittaker index to soft bottom assemblages of the Adriatic Sea highlighted its sensitivity to rare species (species recorded in a single sample). This over-weighting of rare species induces biased estimates of the heterogeneity, thus it becomes difficult to compare assemblages containing a high proportion of rare species. In benthic communities, the unusual large number of rare species is frequently attributed to a combination of sampling errors and insufficient sampling effort. In order to reduce the influence of rare species on the measure of beta diversity, I have developed an alternative index based on simple probabilistic considerations. It turns out that this probability index is an ordinary Michaelis-Menten transformation of Whittaker's index but behaves more favourably when species heterogeneity increases. The suggested index therefore seems appropriate when comparing patterns of complexity in marine benthic assemblages. Although the new index makes an important contribution to the study of biodiversity in sedimentary environment, it remains to be seen which processes, and at what scales, influence benthic patterns. The ability to predict the effects of ecological phenomena on benthic fauna highly depends on both spatial and temporal scales of variation. Once defined, implicitly or explicitly, these scales influence the questions asked, the methodological approaches and the interpretation of results. Problem often arise when representative samples are not taken and results are over-generalized, as can happen when results from small-scale experiments are used for resource planning and management. Such issues, although globally recognized, are far from been resolved in the North Adriatic Sea. This area is potentially affected by both natural (e.g. river inflow, eutrophication) and anthropogenic (e.g. gas extraction, fish-trawling) sources of disturbance. Although few studies in this area aimed at understanding which of these processes mainly affect macrobenthos, these have been conducted at a small spatial scale, as they were designated to examine local changes in benthic communities or particular species. However, in order to better describe all the putative processes occurring in the entire area, a high sampling effort performed at a large spatial scale is required. The sedimentary environment of the western part of the Adriatic Sea was extensively studied in this thesis. I have described, in detail, spatial patterns both in terms of sedimentary characteristics and macrobenthic organisms and have suggested putative processes (natural or of human origin) that might affect the benthic environment of the entire area. In particular I have examined the effect of off shore gas platforms on benthic diversity and tested their effect over a background of natural spatial variability. The results obtained suggest that natural processes in the North Adriatic such as river outflow and euthrophication show an inter-annual variability that might have important consequences on benthic assemblages, affecting for example their spatial pattern moving away from the coast and along a North to South gradient. Depth-related factors, such as food supply, light, temperature and salinity play an important role in explaining large scale benthic spatial variability (i.e., affecting both the abundance patterns and beta diversity). Nonetheless, more locally, effects probably related to an organic enrichment or pollution from Po river input has been observed. All these processes, together with few human-induced sources of variability (e.g. fishing disturbance), have a higher effect on macrofauna distribution than any effect related to the presence of gas platforms. The main effect of gas platforms is restricted mainly to small spatial scales and related to a change in habitat complexity due to a natural dislodgement or structure cleaning of mussels that colonize their legs. The accumulation of mussels on the sediment reasonably affects benthic infauna composition. All the components of the study presented in this thesis highlight the need to carefully consider methodological aspects related to the study of sedimentary habitats. With particular regards to the North Adriatic Sea, a multi-scale analysis along natural and anthopogenic gradients was useful for detecting the influence of all the processes affecting the sedimentary environment. In the future, applying a similar approach may lead to an unambiguous assessment of the state of the benthic community in the North Adriatic Sea. Such assessment may be useful in understanding if any anthropogenic source of disturbance has a negative effect on the marine environment, and if so, planning sustainable strategies for a proper management of the affected area.

Effect of Paramuricea clavata (Risso, 1826) "forest" on the benthic populations of Tremiti Islands Effetto "foresta" di Paramuricea clavata (Risso, 1826) sui popolamenti bentonici delle Isole Tremiti

Mediterranean coralligenous habitats are biogenic reefs characterised by high species diversity and built mainly by encrusting calcareous red algae, growing in dim light conditions. The global climate change and several human activities may threaten species living in these habitats, especially some of those that are considered particularly relevant in structuring and in maintaining the complexity and diversity of the benthic assemblages. Among them, the red gorgonian, Paramuricea clavata (Risso, 1826), which can form dense populations, in the last decades showed worrying mass mortality events. Understand the role of this “animal forests” in the coralligenous assemblages is of fundamental importance in order to design appropriate monitoring programs and conservation policies, especially in the marine protected areas. For this purpose, benthic assemblages were studied in presence and absence of red gorgonians at two sites at the Tremiti islands. Overall, the benthic assemblages significantly differed among sites, nevertheless in both places, clear differences between assemblages associated and not associated to the gorgonian forests were found. In particular, encrusting corallinered algae were significantly more abundant in the gorgonian understories at both sites. This result indicates that the gorgonians may promote the development of calcareous algae, which are the main builders of the coralligenous habitats. Moreover species diversity resulted higher in the assemblages associated to the gorgonians. The present study highlights the role of Paramuricea clavata as a relevant ecosystem engineer in the coralligenous habitats.

Effects of temperature and acidity on Mediterranean benthic calcifiers, in the face of globale climate change

The Mediterranean Sea is expected to react faster to global change compared to the ocean and is already showing more pronounced warming and acidification rates. A study performed along the Italian western coast showed that porosity of the skeleton increases with temperature in the zooxanthellate (i.e. symbiotic with unicellular algae named zooxanthellae) solitary scleractinian Balanophyllia europaea while it does not vary with temperature in the solitary non-zooxanthellate Leptopsammia pruvoti. These results were confirmed by another study that indicated that the increase in porosity was accompanied by an increase of the fraction of the largest pores in the pore-space, perhaps due to an inhibition of the photosynthetic process at elevated temperatures, causing an attenuation of calcification. B. europaea, L. pruvoti and the colonial non-zooxanthellate Astroides calycularis, transplanted along a natural pH gradient, showed that high temperature exacerbated the negative effect of lowered pH on their mortality rates. The growth of the zooxanthellate species did not react to reduced pH, while the growth of the two non-zooxanthellate species was negatively affected. Reduced abundance of naturally occurring B. europaea, a mollusk, a calcifying and a non-calcifying macroalgae were observed along the gradient while no variation was seen in the abundance of a calcifying green alga. With decreasing pH, the mineralogy of the coral and mollusk did not change, while the two calcifying algae decreased the content of aragonite in favor of the less soluble calcium sulphates and whewellite (calcium oxalate), possibly as a mechanism of phenotypic plasticity. Increased values of porosity and macroporosity with CO2 were observed in B. europaea specimens, indicating reduces the resistance of its skeletons to mechanical stresses with increasing acidity. These findings, added to the negative effect of temperature on various biological parameters, generate concern on the sensitivity of this zooxanthellate species to the envisaged global climate change scenarios.

Temporal dynamics of benthic assemblages along a gradient of ocean acidification at a CO2 vent’s system of the Ischia Island

Global warming and ocean acidification, due to rising atmospheric levels of CO2, represent an actual threat to terrestrial and marine environments. Since Industrial Revolution, in less of 250 years, pH of surface seawater decreased on average of 0.1 unit, and is expected to further decreases of approximately 0.3-0.4 units by the end of this century. Naturally acidified marine areas, such as CO2 vent systems at the Ischia Island, allow to study acclimatation and adaptation of individual species as well as the structure of communities, and ecosystems to OA. The main aim of this thesis was to study how hard bottom sublittoral benthic assemblages changed trough time along a pH gradient. For this purpose, the temporal dynamics of mature assemblages established on artificial substrates (volcanic tiles) over a 3 year- period were analysed. Our results revealed how composition and dynamics of the community were altered and highly simplified at different level of seawater acidification. In fact, extreme low values of pH (approximately 6.9), affected strongly the assemblages, reducing diversity both in terms of taxa and functional groups, respect to lower acidification levels (mean pH 7.8) and ambient conditions (8.1 unit). Temporal variation was observed in terms of species composition but not in functional groups. Variability was related to species belonging to the same functional group, suggesting the occurrence of functional redundancy. Therefore, the analysis of functional groups kept information on the structure, but lost information on species diversity and dynamics. Decreasing in ocean pH is only one of many future global changes that will occur at the end of this century (increase of ocean temperature, sea level rise, eutrophication etc.). The interaction between these factors and OA could exacerbate the community and ecosystem effects showed by this thesis.

Metacommunity Dynamics Of Benthic Macroinvertebrates In A Large River

Metacommunity ecology focuses on the interaction between local communities and is inherently linked to dispersal as a result. Within this framework, communities are structured by a combination of in-site responses to the immediate environment (species sorting), stochasticity (patch dynamics), and connections to other communities via distance between communities and dispersal (neutrality), and source-sink dynamics (mass effects; see Chapter 1 for a detailed description of metacommunity theory, the study site, and macroinvertebrate communities found). In Chapter 2 I describe spatial scale of study and dispersal ability as both have the ability to influence the degree to which communities interact. However, little is known about how these factors influence the importance of all metacommunity dynamics. I compared dispersal mode of immature aquatic insects and dispersal ability of winged adults across multiple spatial scales in a large river. The strongest drivers of river communities were patch dynamics, followed by species sorting, then neutrality. Active dispersers during aquatic lifestages on average exhibited lower patch dynamics, higher species sorting, and significant mass effects compared to passive dispersers. Active and strong dispersers also had a scale-independent influence of neutrality, while neutrality was stronger at broader spatial scale for passive and weak dispersers. These results indicate as dispersal ability increases patch dynamics decreases, species sorting increases, and neutrality should decrease. The perceived influence of neutrality may also be dependent on spatial scale and dispersal ability. In Chapter 3 I describe how river benthic macroinvertebrate communities may influence tributary invertebrate communities via adult flight and tributaries may influence mainstem communities via immature drift. This relationship may also depend on relative mainstem and tributary size, as well as abiotic tributary influence on mainstem habitat. To investigate the interaction between a larger river and tributary I sampled mainstem benthic invertebrate communities and quantified habitat of a 7th order river (West Branch Susquehanna River) above and below a 5th order tributary confluence, as well as 0.95-3.2 km upstream in the tributary. Non-metric multidimensional scaling showed similar patterns of clustering between sampling locations for both habitat characteristics and invertebrate communities. In addition, mainstem river communities and habitat directly downstream of the tributary confluence cluster tightly together, intermediate between tributary and mid-channel river samples. In Bray-Curtis dissimilarity comparisons between tributary and mainstem river communities the furthest upstream tributary communities were least similar to river communities. Middle tributary samples were also closest by Euclidean distance to the upstream mainstem riffle and exhibited higher similarity to mid-channel samples than the furthest downstream tributary communities. My results indicate river and tributary benthic invertebrate communities may interact and likely result in direct and indirect mass effects of a tributary on the downstream mainstem community by invertebrate drift and habitat restructuring via material delivery from the tributary. I also showed likely direct effects of adult dispersal from the river and oviposition in proximal tributary locations where Euclidian, rather than river, distance may be more important in determining river-tributary interactions.

Tributary and Mainstem Benthic Macroinvertebrate Communities Linked By Direct Dispersal and Indirect Habitat Alteration

Benthic communities in tributary-mainstem networks might interact via downstream drift of invertebrates or material from tributaries and adult dispersal from the mainstem. Depending on the strength of these interactions, mainstem downstream communities are expected to be more similar to tributary communities due to drift or habitat alteration. Communities not connected by flow are expected to be similar due to adult dispersal but decreasing in similarity with distance from the mainstem. We investigated interactions between invertebrate communities of a 7th order river and 5th order tributary by comparing benthic community structure in the river upstream and downstream of the tributary confluence and upstream in the tributary. Non-metric multidimensional scaling showed invertebrate communities and habitat traits from river locations directly downstream of the tributary clustered tightly, intermediate between tributary and mid-channel river locations. In addition, Bray-Curtis dissimilarity increased between the mainstem and tributary with distance upstream in the tributary. Our results indicate that similarities between mainstem and tributary communities are potentially caused by direct mass effects from tributary to downstream mainstem communities by invertebrate drift and indirect mass effects by habitat restructuring via material delivery from the tributary, as well as potential effects of adult dispersal from the river on proximal tributary communities.

Hidden Biodiversity in an Ecologically Important Freshwater Amphipod: Differences in Genetic Structure between Two Cryptic Species

Cryptic species, i.e. species that are morphologically hard to distinguish, have been detected repeatedly in various taxa and ecosystems. In order to evaluate the importance of this finding, we have to know in how far cryptic species differ in various aspects of their biology. The amphipod Gammarus fossarum is a key invertebrate in freshwater streams and contains several cryptic species. We examined the population genetic structure, genetic diversity and demographic history of two of them (type A and type B) using microsatellite markers and asked whether they show significant differences. We present results of population genetic analyses based on a total of 37 populations from the headwaters of two major European drainages, Rhine and Rhone. We found that, in both species, genetic diversity was geographically structured among and within drainages. For type A in the Rhine and type B in the Rhone, we detected significant patterns of isolation by distance. The increase of genetic differentiation with geographical distance, however, was much higher in type A than in type B. This result indicates substantial interspecific differences in population history and/or the extent of current gene flow between populations. In the Rhine, type B does not show evidence of isolation by distance, and population differentiation is relatively low across hundreds of kilometres. The majority of these populations also show signatures of recent bottlenecks. These patterns are consistent with a recent expansion of type B into the Rhine drainage. In summary, our results suggest considerable and previously unrecognized interspecific differences in the genetic structure of these cryptic keystone species.