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Viruses are the most abundant 'biological entities' in the world's oceans. However, technical and methodological constraints limit our understanding of their diversity, particularly in benthic abyssal ecosystems (>4000 m depth). To verify advantages and limitations of analyzing virome DNA subjected either to random amplification or unamplified, we applied shotgun sequencing-by-synthesis to two sample pairs obtained from benthic abyssal sites located in the North-eastern Atlantic Ocean at ca. 4700 m depth. One amplified DNA sample was also subjected to single-molecule long-read sequencing for comparative purposes. Overall, we identified 24,828 viral Operational Taxonomic Units (vOTUs), belonging to 22 viral families. Viral reads were more abundant in the amplified DNA samples (38.5-49.9%) compared to the unamplified ones (4.4-5.8%), with the latter showing a greater viral diversity and 11-16% of dsDNA viruses almost undetectable in the amplified samples. From a procedural point of view, the viromes obtained by direct sequencing (without amplification step) provided a broader overview of both ss and dsDNA viral diversity. Nevertheless, our results suggest that the contextual use of random amplification of the same sample and long-read technology can improve the assessment of viral assemblages by reducing off-target reads.

Amidst a global biodiversity crisis 1 , the word 'biodiversity' has become indispensable for conservation and management 2 . Yet, biodiversity is often used as a buzzword in scientific literature. Resonant titles of papers claiming to have studied 'global biodiversity' may be used to promote research focused on a few taxonomic groups, habitats, or facets of biodiversity - taxonomic, (phylo)genetic, or functional. This usage may lead to extrapolating results outside the target systems of these studies with direct consequences for our understanding of life on Earth and its practical conservation. Here, we used a random sample of papers with the word 'biodiversity' in their title to take a long view of the use of this term. Despite improvements in analytical tools, monitoring technologies, and data availability 3 , 4 , we found that the taxonomic scope of research articles has not increased in recent years. We also show that studies with a wider taxonomic scope attract more citations and online attention. Our results have broad ramifications for understanding how extrapolating from studies with narrow taxonomic scope affects our view of global biodiversity and conservation., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Coastal tourism is a growing industry sector in the Mediterranean Basin. This and the other human activities occurring along the coastline share space and resources, leading to conflicts for divergent uses. Moreover, the overexploitation of natural resources degrades and depletes coastal habitats, with negative feedback effects for all human activities. Hence, both tourism and the other human activities have to consider their dependence on coastal ecosystem services, and act at technical and policy level to reach a compromise that preserves natural resources in the long term. Here we provide a conceptual framework illustrating the complex relationships and trade-offs among threats from coastal tourism and from other human activities and coastal ecosystem services, with a focus on cultural ones. We discuss the negative feedbacks on tourism development and provide examples of geospatial analysis on cumulative threats generated by other human activities and affecting tourism itself. The proposed conceptual framework and the threat analysis aim at highlighting the negative feedback effects of human driven threats on the development of Mediterranean coastal tourism, through an ecosystem service perspective. Both tools provide valuable insight for supporting decision makers and planners in achieving integrated coastal management, with a focus on sustainable tourism., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Estuaries are known to export huge quantities of terrigenous/riverine organic matter and nutrients to coastal areas, subsidizing food sources to their communities; however, this process is seldom investigated in complex multichannel delta-prodelta systems. Stable isotope analysis was used to investigate the contribution of organic sources originated from the Po River Delta, one of the widest deltaic wetlands in the northern Mediterranean Sea, to the diet of the nearby coastal macrobenthic consumers, and their influence on invertebrates' trophic structure and biomass. Following intense river flood events, macrofauna samples were collected in the prodelta area at increasing distance from the main river distributary mouth. Potential primary resources were collected within the delta and the prodelta areas. A terrestrial signal in the primary consumers' diet was evident as a shift in their δ 13 C, being this more 13 C depleted near the main river distributary. The Bayesian mixing models indicated an important contribution of deltaic sources, in particular C4-grass detritus (25-57%), to primary consumers' diet, confirming a strong energetic link between delta and prodelta areas. Overall, most of the consumers' biomass were concentrated at sites close to the main distributary mouth, mainly as suspension and surface deposit feeders. A simplification of the macrobenthic community structure, accompanied by narrower transfer pathways was also evident at sites more distant from the main river distributary, in relation to changes in the quality of resources. Our data, although limited to winter season, suggest that during periods of low in situ productivity but high river flow energy, invertebrates are able to efficiently exploit terrigenous food sources, restraining the detrimental effect of increased turbidity and sedimentation. We conclude that riparian/wetland vegetation associated with river deltas can provide important food sources to marine primary consumers. These results furthermore highlight the need for integrated management and protection strategies of connected land-sea ecosystems., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Dense waters (DW) formation in shelf areas and their cascading off the shelf break play a major role in ventilating deep waters, thus potentially affecting ecosystem functioning and biogeochemical cycles. However, whether DW flow across shelves may affect the composition and structure of plankton communities down to the seafloor and the particles transport over long distances has not been fully investigated. Following the 2012 north Adriatic Sea cold outbreak, DW masses were intercepted at ca. 460 km south the area of origin and compared to resident ones in term of plankton biomass partitioning (pico to micro size) and phytoplankton species composition. Results indicated a relatively higher contribution of heterotrophs in DW than in deep resident water masses, probably as result of DW-mediated advection of fresh organic matter available to consumers. DWs showed unusual high abundances of Skeletonema sp., a diatom that bloomed in the north Adriatic during DW formation. The Lagrangian numerical model set up on this diatom confirmed that DW flow could be an important mechanism for plankton/particles export to deep waters. We conclude that the predicted climate-induced variability in DW formation events could have the potential to affect the ecosystem functioning of the deeper part of the Mediterranean basin, even at significant distance from generation sites.

Growing evidence indicates that dense water formation and flow over the continental shelf is a globally relevant oceanographic process, potentially affecting microbial assemblages down to the deep ocean. However, the extent and consequences of this influence have yet to be investigated. Here it is shown that dense water propagation to the deep ocean increases the abundance of prokaryotic plankton, and stimulates carbon production and organic matter degradation rates. Dense waters spilling off the shelf modifies community composition of deep sea microbial assemblages, leading to the increased relevance of taxa likely originating from the sea surface and the seafloor. This phenomenon can be explained by a combination of factors that interplay during the dense waters propagation, such as the transport of surface microbes to the ocean floor (delivering in our site 0.1 megatons of C), the stimulation of microbial metabolism due to increased ventilation and nutrients availability, the sediment re-suspension, and the mixing with ambient waters along the path. Thus, these results highlight a hitherto unidentified role for dense currents flowing over continental shelves in influencing deep sea microbes. In light of climate projections, this process will affect significantly the microbial functioning and biogeochemical cycling of large sectors of the ocean interior., (© 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Carbon and nitrogen stable isotope analysis was used to examine the food web of the Mar Piccolo of Taranto, a coastal basin experiencing several anthropogenic impacts. Main food sources (algal detritus, seaweeds, particulate organic matter (POM) and sediment organic matter (SOM)) and benthic and pelagic consumers were collected during two contrasting seasons (June and April), at four sites distributed over two inlets, and characterized by different level of confinements, anthropogenic inputs and the presence of mussels farming. δ(13)C values of organic sources revealed an important contribution of POM to both planktonic and benthic pathways, as well as the influence of terrigenous inputs within both inlets, probably due to high seasonal land runoff. Although δ(13)C of both sources and consumers varied little between sampling sites and dates, δ(15)N spatial variability was higher and clearly reflected the organic enrichment in the second inlet as well as the uptake of anthropogenically derived material by benthic consumers. On the other hand, within the first inlet, the isotopic composition of consumers did not change in response to chemical contamination. However, the impact of polluted sediments near the Navy Arsenal in the first inlet was detectable at the level of the macrobenthic trophic structure, showing high dominance of motile, upper level consumers capable to face transient conditions and the reduction of the more resident deposit feeders. We therefore underline the great potential of matching stable isotope analysis with quantitative studies of community structure to assess the effects of multiple anthropogenic stressors.

Knowledge on ecosystem functioning can largely contribute to promote ecosystem-based management and its application. The Mar Piccolo of Taranto is a densely populated area at a high risk of environmental crisis. Here, planktonic primary production (PP) and heterotrophic prokaryotic production (HPP) were measured as proxies of functioning in three sampling sites located in two inlets at different levels of industrial contamination, during three sampling surveys (June 2013, February and April 2014). To have a better overall view and provide some insights into the benthic-pelagic coupling, we integrated PP and HPP in the water column with those in the sediments and then discussed this with the origin of the organic matter pools based on analysis of stable isotopes. Heavy metals and polychlorobiphenyls (PCBs) were also analysed in the surface (1 cm) sediment layer and related to the overall ecosystem functioning. Multidimensional scaling (MDS) analysis, based on the main data, clearly separated the second inlet from the first one, more severely impacted by anthropogenic activities. The stable isotope mixing model suggested the prevalent terrestrial/riverine origin of the particulate organic matter pools (mean 45.5 %) in all sampling periods, whereas phytoplankton contributed up to 29 % in February. Planktonic PP and HPP rates followed the same pattern over the entire study period and seemed to respond to phytoplankton dynamics confirming this community as the main driver for the C cycling in the water column. On the contrary, benthic PP rates were almost negligible while HPP rates were lower or comparable to those in the water column indicating that although the Mar Piccolo is very shallow, the water column is much more productive than the surface sediments. The sediment resuspension is likely responsible for a pulsed input of contaminants into the water column. However, their interference with the proper functioning of the pelagic ecosystem seems to be limited to the bottom layers.

Background: Primin is the main contact allergen found in the ornamental plant Primula obconica Hance., Objective: To analyse the temporal trend of sensitivity to primin in north-eastern Italy and to evaluate the associations with occupations in our geographical area., Methodology: From 1996 to 2012, 24 052 consecutive patients with suspected allergic contact dermatitis were patch tested in north-eastern Italy. Individual characteristics were collected through a standardized questionnaire in eight departments of dermatology or occupational medicine., Results: The overall prevalence of primin sensitization was 1.9%; the prevalence was significantly higher in women (2.6%) than in men (0.5%). The Pordenone area had the higher prevalence of sensitization, which reached 6% in 1999-2001. We found a significant association between primin sensitization and household workers [odds ratio (OR) 2.3; 95% confidence interval (CI): 1.61-3.35], retired people (OR 1.8; 95%CI: 1.22-2.81), woodworkers (OR 2.1; 95%CI: 1.10-6.18), and chemical industry workers (OR 2.9; 95%CI: 1.05-8.29)., Conclusion: Our study showed that contact allergy to primin is still relevant in north-eastern Italy. The frequency of sensitization is decreasing, but for retired and household workers it is still >4%. Our results suggest the need to promote the use of primin-free P. obconica in Italy., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Several seamounts are known as 'oases' of high abundances and biomass and hotspots of biodiversity in contrast to the surrounding deep-sea environments. Recent studies have indicated that each single seamount can exhibit a high intricate habitat turnover. Information on alpha and beta diversity of single seamount is needed in order to fully understand seamounts contribution to regional and global biodiversity. However, while most of the seamount research has been focused on summits, studies considering the whole seamount structure are still rather poor. In the present study we analysed abundance, biomass and diversity of nematodes collected in distinct physiographic sites and surrounding sediments of the Condor Seamount (Azores, North-East Atlantic Ocean). Our study revealed higher nematode biomass in the seamount bases and values 10 times higher in the Condor sediments than in the far-field site. Although biodiversity indices did not showed significant differences comparing seamount sites and far-field sites, significant differences were observed in term of nematode composition. The Condor summit harboured a completely different nematode community when compared to the other seamount sites, with a high number of exclusive species and important differences in term of nematode trophic diversity. The oceanographic conditions observed around the Condor Seamount and the associated sediment mixing, together with the high quality of food resources available in seamount base could explain the observed patterns. Our results support the hypothesis that seamounts maintain high biodiversity through heightened beta diversity and showed that not only summits but also seamount bases can support rich benthic community in terms of standing stocks and diversity. Furthermore functional diversity of nematodes strongly depends on environmental conditions link to the local setting and seamount structure. This finding should be considered in future studies on seamounts, especially in view of the potential impacts due to current and future anthropogenic threats.

Deep-sea fishes provide a unique opportunity to study the physiology and evolutionary adaptation to extreme environments. We carried out a high throughput sequencing analysis on a 454 GS-FLX titanium plate using unnormalized cDNA libraries from six tissues of A. carbo. Assemblage and annotations were performed by Newbler and InterPro/Pfam analyses, respectively. The assembly of 544,491 high quality reads provided 8,319 contigs, 55.6% of which retrieved blast hits against the NCBI nonredundant database or were annotated with ESTscan. Comparison of functional genes at both the protein sequences and protein stability levels, associated with adaptations to depth, revealed similarities between A. carbo and other bathypelagic fishes. A selection of putative genes was standardized to evaluate the correlation between number of contigs and their normalized expression, as determined by qPCR amplification. The screening of the libraries contributed to the identification of new EST simple-sequence repeats (SSRs) and to the design of primer pairs suitable for population genetic studies as well as for tagging and mapping of genes. The characterization of the deep-sea fish A. carbo first transcriptome is expected to provide abundant resources for genetic, evolutionary, and ecological studies of this species and the basis for further investigation of depth-related adaptation processes in fishes.

Decomposition of organic matter in marine sediments is a critical step influencing oxygen and carbon fluxes. In addition to heterotrophic bacteria and fungi, osmoheterotrophic protists may contribute to this process, but the extent of their role as decomposers is still unknown. Among saprophytic protists, the thraustochytrids have been isolated from different habitats and substrates. Recently, they have been reported to be particularly abundant in marine sediments characterized by the presence of recalcitrant organic matter such as seagrass and mangrove detritus where they can reach biomass comparable to those of other protists and bacteria. In addition, their capacity to produce a wide spectrum of enzymes suggests a substantial role of thraustochytrids in sedimentary organic decomposition. Moreover, thraustochytrids may represent a food source for several benthic microorganisms and animals and may be involved in the upgrading of nutrient-poor organic detritus. This chapter presents an overview on studies of thraustochytrids in benthic ecosystems and discusses future prospectives and possible methods to quantify their role in benthic food webs.

The object of this paper is to describe the imaging and clinical characteristics of subependymal nodule (SN) - subependymal giant cell astrocytoma (SGCA) complex in tuberous sclerosis and analyze its evolution in order to attempt early detection and the prevention of intracranial hypertension. We evaluated 22 patients with the pathological diagnosis of SGCA. The diagnosis was made at a median of 10.1 years old. We were able to observe the evolution of SN to ASGC: these SN were localized adjacent to the foramen of Monro and with time they underwent an important development with intense contrast enhancement and hydrocephalus. The acceleration in SN growth and its "transformation" into SGCA occurred at an average of 10 years of age, with a mean diameter of 9 mm. No SN located far from the foramen of Monro evolutioned to SGCA. Fifteen patients (68%) were operated with symptoms of intracranial hypertension. Average age at surgery was 10.8 years old. Six patients presented visual deficit and in these, the average diameter of the tumor was 31.5 mm, a high value when compared to 18.7 mm in the patients without visual deficit. The imaging and clinical follow-up of any subependymal lesion close to the foramen of Monro will permit, at a presymptomatic stage, an anticipation of surgical treatment thus reducing intracranial hypertension incidence. Prospective studies could determine whether the SN-SGCA complex corresponds to the same entity in distinct evolution stages or to two lesions with different growth potential.

Purpose: We studied the electroclinical features and evolution in patients with childhood absence epilepsy (CAE) associated with electroencephalographic findings similar to those of benign focal epilepsies (BFE) with or without clinical manifestations compatible with these focal idiopathic syndromes., Methods: Between June 1994 and June 2002, we found 203 (3.6%) patients with typical electroclinical features of CAE among 8285 children with epilepsy. From this population of 203, we found 30 cases (14.7%) that also showed focal abnormalities of BFE on the EEG. Seven of these 30 cases also had clinical manifestations of BFE that preceded the onset of the absences., Results: There were 20 (66.5%) boys and 10 (33.5%) girls. Age at onset of absences ranged from 2 to 10.5 years, with a mean age of 5.5 years. Of 30, 7 had focal clinical seizures as well. Three of seven had seizures characteristic of Panayiotopoulos syndrome (PS), and the other four had seizures compatible with childhood occipital epilepsy (COE) of Gastaut. The focal seizures started between 3 and 7 years of age. In all patients seizures were under control within 2-24 months (mean: 11 months) after onset. The focal discharges disappeared in 26 patients at a mean age of 8 years (range 4-13 years), 1 year after the typical absences had disappeared. In four patients the focal paroxysms are still present., Conclusion: The association of two different idiopathic focal and generalized epilepsies in the same patient may be merely coincidental, but a close genetic relationship between both epileptic syndromes might be another hypothesis. Another explanation could be that our series of patients represent a subgroup of CAE.

Purpose: We present a series of nine patients with early-onset hydrocephalus who had seizures and continuous spikes and waves during slow sleep (CSWS) associated with neurocognitive and motor deterioration., Methods: Six boys and three girls aged 9-16 years (mean 11.3 years) were studied. [Correction added after online publication 12-Apr-2008: Number of girls and boys has been updated.] All patients underwent clinical examinations, electroencephalographic evaluations, neuroradiological imaging and neuropsychological assessment at first examination. Antiepileptic drugs (AEDs) were given in all cases and changed according to clinical and EEG evolution., Results: Onset of epilepsy occurred from age 8 to 60 months (mean 19.6 months and median 14 months) with focal seizures with or without secondary generalized tonic-clonic seizures. Between ages 6 and 13 years (mean 10.4 years and median 8 years), hyperkinesia, aggressiveness, and poor socialization appeared in all nine cases. Reduced attention span, deterioration of language, and temporospatial disorientation were found in three of them. Negative myoclonus was found in two patients. The EEG showed CSWS. Response to change in treatment was good in all patients. None of the patients had relapses, seven of them have remained seizure free, and two continued having sporadic focal motor seizures during 2-5 years (mean 3 years) of follow-up., Conclusion: In children with early-onset hydrocephalus, particularly with behavioral and language disturbances and/or motor deterioration, CSWS should be considered. Periodic EEG recordings during sleep should be done in these children. The early identification of this particular electroclinical picture is crucial to start adequate treatment to avoid progressive cognitive deterioration.

Background: Coral bleaching (i.e., the release of coral symbiotic zooxanthellae) has negative impacts on biodiversity and functioning of reef ecosystems and their production of goods and services. This increasing world-wide phenomenon is associated with temperature anomalies, high irradiance, pollution, and bacterial diseases. Recently, it has been demonstrated that personal care products, including sunscreens, have an impact on aquatic organisms similar to that of other contaminants., Objectives: Our goal was to evaluate the potential impact of sunscreen ingredients on hard corals and their symbiotic algae., Methods: In situ and laboratory experiments were conducted in several tropical regions (the Atlantic, Indian, and Pacific Oceans, and the Red Sea) by supplementing coral branches with aliquots of sunscreens and common ultraviolet filters contained in sunscreen formula. Zooxanthellae were checked for viral infection by epifluorescence and transmission electron microscopy analyses., Results: Sunscreens cause the rapid and complete bleaching of hard corals, even at extremely low concentrations. The effect of sunscreens is due to organic ultraviolet filters, which are able to induce the lytic viral cycle in symbiotic zooxanthellae with latent infections., Conclusions: We conclude that sunscreens, by promoting viral infection, potentially play an important role in coral bleaching in areas prone to high levels of recreational use by humans.

Although the relationships between trophic conditions and viral dynamics have been widely explored in different pelagic environments, there have been few attempts at independent estimates of both viral production and decay. In this study, we investigated factors controlling the balance between viral production and decay along a trophic gradient in the north Adriatic basin, providing independent estimates of these variables and determining the relative importance of nanoflagellate grazing and viral life strategies. Increasing trophic conditions induced an increase of bacterioplankton growth rates and of the burst sizes. As a result, eutrophic waters displayed highest rates of viral production, which considerably exceeded observed rates of viral decay (up to 2.9 x 10(9) VLP liter(-1) h(-1)). Viral decay was also higher in eutrophic waters, where it accounted for ca. 40% of viral production, and dropped significantly to 1.3 to 10.7% in oligotrophic waters. These results suggest that viral production and decay rates may not necessarily be balanced in the short term, resulting in a net increase of viruses in the system. In eutrophic waters nanoflagellate grazing, dissolved-colloidal substances, and lysogenic infection were responsible together for the removal of ca. 66% of viral production versus 17% in oligotrophic waters. Our results suggest that different causative agents are primarily responsible for the removal of viruses from the water column in different trophic conditions. Factors other than those considered in the past might shed light on processes responsible for the removal and/or decay of viral particles from the water column.

Thraustochytrids are ubiquitous, chemo-organotrophic, marine stramenipilan protists belonging to the class Labyrinthulomycetes. Their taxonomy is largely based on life cycle development stages. We describe here a new species of thraustochytrid isolated from mangroves of Goa, India. The organism is characterized by large zoosporangia with two distinct development cycles. In one, typical thalli with ectoplasmic net elements mature into zoosporangia that divide to form heterokont biflagellate zoospores, leaving behind a proliferation body. In the second type, the thalli develop into amoeboid cells, reminiscent of the genus Ulkenia Gaertner. Unlike Ulkenia, however, the 'amoebae' do not immediately produce zoospores, but round up prior to division into zoospores. The two types of development occur simultaneously in single cell-derived in- vitro cultures. Molecular characterization of the new isolate involving 18S rRNA gene typing and comparative phylogenetic analysis further establish it to be a new and distinct thraustochytrid species with Schizochytrium aggregatum Goldstein and Belsky and Thraustochytrium kinnei Gaertner as the closest forms. We have named this new species as Thraustochytrium gaertnerium, deriving its species name in honour of Dr Alwin Gaertner, a pioneer in the studies of taxonomy and ecology of thraustochytrids.

We investigated the impact of fish farm biodeposition on benthic bacteria, thraustochytrid protists, and heterotrophic protozoa (nanoflagellates and ciliates) in an oligotrophic area of the Mediterranean Sea. The fish farm impact was investigated both on a seagrass (Posidonia oceanica) bed and on soft bottom sediments. In both systems, sediment samples were collected with a multicontrol sampling strategy (i.e., beneath the fish farm and at three control sites per system). The uneaten food pellets supplied to the fish determined the accumulation of sediment organic matter and the enhancement of protein content in impacted sediments (both seagrass bed and soft sediments). In both systems, the abundance and biomass of heterotrophic protists increased significantly beneath the fish farm, but the structure of the protist assemblages responded differently in vegetated and unvegetated sediments. Thraustochytrid abundance increased significantly in impacted seagrass. These results provide evidence that the structure of protist assemblages respond significantly to fish farm biodeposition and indicate that the monitoring of these benthic components provides complementary information for the assessment of the fish farm impact on the benthic systems.

The appearance of pink-blue spots (termed here as pink-blue spot syndrome - PBSS) in the branching coral Acropora eurystoma from the Gulf of Eilat, Red Sea, is described. We monitored 18 transects (10 x 1 m2 each) in front of the H. Steinitz Marine Laboratory (Eilat), at 3, 6 and 9 m depth, during March and August in 2001 and 2002. Transect measurements revealed high frequencies of colonies with PBSS (up to 100% of colonies) between 3 and 9 m depth. Ten PBSS-affected colonies of A. eurystoma were labelled and monitored for the development of spots. From March to August 2001, the number of spots per colony increased and remained constantly high at both sampling dates in 2002. Spot size ranged between 7 and 149 mm2. Spots were primarily recorded in areas where coral tissues contacted foreign biological matter, either around regenerative wounds or when surrounded by encrusting organisms, in fast-growing areas and in allogeneic interactions. A preliminary biochemical examination suggested that the pink-blue pigment in A. eurystoma is part of a family of compounds (pocilloporin) responsible for the pink-blue colours in pocilloporid and acroporid corals. Pink-blue colour could be experimentally induced in A. eurystoma by tissue-to-tissue contacts between distressed and non-distressed allogeneic branches. PBSS was also induced in healthy coral tissue by contact with inert objects, e.g., by bandaging a branch with plastic strips. Any specific pink-blue colour spots faded within 1-3 months from onset. These results suggest that PBSS in A. eurystoma may not be considered a regular coral disease, but rather a locally induced syndrome caused by restricted environmental and/or biological stress conditions.

Background: The aim of this study was to evaluate the incidence of major amputations and critical limb ischemia in an unselected population of western Tuscany., Methods: Out of an overall 1234000 people living in Pisa and surroundings (western Tuscany), a total amount of 306 patients underwent 319 different major/minor amputations during the year 2002 in private and public hospitals, both in Tuscany and in other regions., Results: 84.6% (n = 270) of the amputations were atherosclerotic, while 15.4% (n = 49) were non-atherosclerotic. The patients with unreconstructable critical limb ischemia, who underwent amputations at the symptomatic limbs, had a minor amputation rate (at toes or forefoot) of 46.7%, while the major amputation rate (over the ankle) was 53.3%. The 144 major amputations were performed in 76.4% (n = 110) of the cases above the knee and in the remaining 23.6% (n = 34) of cases below the knee. Early hospital mortality rates of the amputees for ischemic causes were 7.6% in patients who underwent major amputations, and 0.8% in cases with minor amputations, respectively. Overall, in western Tuscany, the incidence of the major amputation rate per million inhabitants was 117 cases per year, with a total of 468 new cases of reconstructable and unreconstructable critical limb ischemia per year (this number was calculated multiplying by 4 the number of major amputations)., Conclusions: On the basis of our real epidemiological data, the theoretical major amputation rates per year were 408 in Tuscany and 6652 in Italy, respectively. The extrapolation of our results showed that the overall critical limb ischemia incidence consists of 1638 cases in Tuscany and of 26676 patients in Italy, respectively.

This field study aims to determine whether increased levels of organically enriched particulate matter released by net pen fish farms (Eilat, Red Sea) would affect the growth of nubbins taken from the branching coral Stylophora pistillata. We followed the survival and growth of 1322 nubbins pruned from five colonies that were transplanted at a depth of 6 m in the vicinity of the fish cages and in a reference site, in front of the Interuniversity Institute (IUI). Nubbins were attached on U-shaped PVC plates in three orientations (up, vertical and down positions). After 50 days, survival was high in both localities and no difference was recorded between the spatial orientations. At the fish farm, however, burial of the nubbin's lateral growths and partial coverage of nubbins by settled particulate matter resulted in significant reduction of the lateral growth rates of nubbins settled in the up position as compared to the reference site. On the other hand, faster growth rates were recorded in the vertical set of nubbins at the fish farm when compared with the IUI site. These results strengthen the conclusion that physical effects, rather than nutrient enrichment, may constitute the main cause of stress for minute coral fragments (resembling coral recruits) growing near the vicinity of a fish farm.