Impact of aquaculture on benthic virus-prokaryote interactions in the Mediterranean Sea.

We investigated the effects of organic enrichment due to the biodeposition from fish farms on benthic prokaryotic and viral abundance and production, viral-induced prokaryotic mortality, enzymatic activities and bacterial diversity. We compared four areas across the Mediterranean Sea, from Cyprus to Spain, and two different habitats: sediments covered by the seagrass Posidonia oceanica and soft-bottom unvegetated sediments. In several cases, the sediments beneath the cages showed higher prokaryotic and viral abundance and production, and higher rates of organic matter decomposition. However, the differences between impact and control sediments were not consistent at all regions and habitats. Benthic bacterial diversity was always lower below the cages, where high viral-induced bacterial mortality rates were also observed. The δ- and γ-Proteobacteria dominated in both impacted and control sediments, but the relative importance of sulphate-reducing δ-Proteobacteria increased beneath the cages. We conclude that aquaculture can have a significant impact on benthic prokaryotes and viruses, by stimulating prokaryotic metabolism and viral infections, reducing bacterial diversity and altering assemblage composition. However, these impacts vary depending upon the sediment type and the habitat characteristics.

Impacts of marine aquaculture at large spatial scales: evidences from N and P catchment loading and phytoplankton biomass.

While several studies point at off-shore aquaculture as a possible source of impacts on the local marine environment, very few have analysed its effects at large scales such as at the bay, gulf or basin levels. Similar analyses are hampered by the multiple sources of disturbance that may concomitantly affect a given area. The present paper addresses these issues taking the Gulf of Castellammare (Southern Tyrrhenian Sea) as an example. Nitrogen (N) and phosphorous (P) loads were calculated for the period 1970-2007, and compared to chlorophyll-a concentration as measured inside and outside the Gulf over the same period. Results indicate that N and P catchment loading has constantly decreased because of improved environmental management. Nevertheless, nutrient concentration in the Gulf has steadily increased since the establishment of aquaculture facilities in 1999. Chlorophyll-a concentration followed this trend, showing a marked increase from 2001 onwards. In the same period, chlorophyll-a concentrations measured inside and outside the Gulf have significantly diverged. As all the other possible causes can be ruled out, aquaculture remains the sole explanation for the observed situation. This paper demonstrates for the first time ever that off-shore aquaculture may affect the marine ecosystem well beyond the local scale and provides an additional element of concern to be kept into consideration when allocating oceans' space for new fish-farming activities.

A structured training program for awake fiber optic intubation: teaching the complete package.

BACKGROUND: We organized a training program for oral fiber optic intubation (FOI) under conscious sedation. The efficacy of the program was evaluated by comparing the performances of experts and novices. METHODS: The training procedure was divided into two sessions: a theoretical session on difficult airways, the fiber optic bronchoscope (FOB), remifentanil, topical anesthesia and patient interactions; and a session involving simulations of the FOI technique on dummies. For in vivo FOI, we enrolled patients requiring orotracheal intubation for elective surgery. Electrocardiograms, mean arterial pressure (MAP) values, peripheral O(2) saturation (SpO(2)) values, respiratory rates (RR) and sedation scores (OAA/S) were acquired. Remifentanil infusion was started at 0.05-0.1 microg*kg(-1)*min(-1), and patients' upper airways were anesthetized with lidocaine. Remifentanil was titrated to achieve an OAA/S of 9-12. FOBs were inserted, and topical laryngeal anesthesia was achieved ("spray as you go" technique). The instrument was passed into the trachea, the OT tube was railroaded over the fiberscope, and tracheal intubation was completed. The procedure duration and patients' vital parameters and satisfaction were recorded. RESULTS: Three experts and four less-experienced anesthesiologists who performed 29 (10, 10 and 9) and 25 (6, 6, 6 and 7) FOIs, respectively, joined the study. To reach the target OAA/S, the remifentanil dosage was progressively increased to an average dose of 0.15+/-0.05 microg*Kg(-1)*min(-1). MAP and SpO(2) values were stable throughout the procedures, HR was slightly increased (from 77+/-16 to 90+/-23 bpm, P=0.02), and RR was decreased (from 16+/-3 to 12+/-4 bpm, P<0.05). No differences were recorded between the experts and less-experienced anesthesiologists. The average duration of FOI was 3.3+/-2.0 min for experts and 4.2+/-2.4 min for novices (P=0.03). Procedures were successful in both groups, with patients in each group being equally satisfied with the procedures. CONCLUSION: This study highlights the importance of a structured FOI training program, demonstrating that it is possible to learn to perform FOI proficiently by practicing on dummies.

Extraintestinal Escherichia coli carrying virulence genes in coastal marine sediments.

Despite the recognized potential of long-term survival or even growth of fecal indicators bacteria (FIB) in marine sediments, this compartment is largely ignored by health protection authorities. We conducted a large-scale study over approximately 50 km of the Marche coasts (Adriatic Sea) at depths ranging from 2 to 5 m. Total and fecal coliforms (FC) were counted by culture-based methods. Escherichia coli was also quantified using fluorescence in situ hybridization targeting specific 16S rRNA sequences, which yielded significantly higher abundances than culture-based methods, suggesting the potential importance of viable but nonculturable E. coli cells. Fecal coliforms displayed high abundances at most sites and showed a prevalence of E. coli. FC isolates (n = 113) were identified by API 20E, additional biochemical tests, and internal transcribed spacer-PCR. E. coli strains, representing 96% of isolates, were then characterized for genomic relatedness and phylogenetic group (A, B1, B2, and D) of origin by randomly amplified polymorphic DNA and multiplex-PCR. The results indicated that E. coli displayed a wide genotypic diversity, also among isolates from the same station, and that 44 of the 109 E. coli isolates belonged to groups B2 and D. Further characterization of B2 and D isolates for the presence of 11 virulence factor genes (pap, sfa/foc, afa, eaeA, ibeA, traT, hlyA, stx(1), stx(2), aer, and fyuA) showed that 90% of B2 and 65% of D isolates were positive for at least one of these. Most of the variance of both E. coli abundance and assemblage composition (>62%) was explained by a combination of physical-chemical and trophic variables. These findings indicate that coastal sediments could represent a potential reservoir for commensal and pathogenic E. coli and that E. coli distribution in marine coastal sediments largely depends upon the physical and trophic status of the sediment. We conclude that future sampling designs aimed at monitoring the microbiological quality of marine coastal areas should not further neglect the analysis of the sediment and that monitoring of these environments can be improved by including molecular methods as a complement of culture-based techniques.

Exo-enzymatic activities and dissolved organic pools in relation with mucilage development in the Northern Adriatic Sea.

We tested the hypothesis that the appearance of mucilage in the Northern Adriatic Sea was related with the accumulation of dissolved organic compounds released by intensive enzymatic activities and not utilized as direct substrate for microbial growth. To do this enzymatic activities and dissolved organic and inorganic pools in periods characterized by the presence of mucilage and in the same seasons but in absence of mucilage were compared. Extracellular enzymatic activities (aminopeptidase, beta-glucosidase and alkaline phosphatase), nutrient pool concentrations (total dissolved nitrogen, dissolved organic nitrogen, total dissolved phosphorus, dissolved organic phosphorus) and the biochemical composition of particulate and dissolved organic matter (in terms of proteins and carbohydrates) were determined on a monthly basis over a period of 3 years. Aminopeptidase and alkaline phosphatase activities displayed higher values in springs preceding the appearance of mucilage than in spring when no mucilage was observed. Beta-Glucosidase activity showed significantly higher values in summer periods characterized by the massive production of mucilage than in summers without mucilage events. The months preceding mucilage events were also characterized by an increase of the alkaline phosphatase to aminopeptidase activity ratio and by a significant accumulation of dissolved proteins. These findings, together with the significant increase of the DON/DOP ratio, suggest that mucilage formation is favoured by the deficiency of organic P. The present study provides compelling evidences that mucilage formation is favoured by the unbalance between organic matter mobilization by enzymatic activities and the accumulation of labile dissolved organic-N compounds.

Assessing the trophic state and eutrophication of coastal marine systems: a new approach based on the biochemical composition of sediment organic matter.

We used a biochemical approach based on the analysis of the quality and quantity of sedimentary organic matter for identifying new descriptors of the trophic state and environmental quality of coastal marine systems. A large-scale study, including 99 stations, belonging to 33 transects, was carried out along 250 km of the Apulian coasts (Mediterranean Sea) in March and September 2000. The investigated area covered a wide range of anthropogenic impacts (industrial ports, tourist harbours, areas affected by power plants and industrial wastes, mariculture areas). Other sites, including marine protected areas (i.e., without any apparent impact), were used as "control". Water column and benthic parameters provided different indications and classifications of the trophic state of coastal marine systems. We found that phytopigment content of the sediments changed in response to all different sources of anthropogenic impact and resulted in a useful descriptor of the trophic state and environmental quality. Highest sediment chlorophyll-a concentrations, indicating conditions of increasing eutrophication, were found in areas impacted by the discharge of heated waters from a power plant. In particular, the contribution of the autotrophic biomass to the biopolymeric carbon pool appeared to be a good descriptor of the decreasing environmental quality. Independently from the sampling period or the pollution source such contribution was significantly lower in transects subjected to anthropogenic impact than in control areas. Differences in trophic conditions were evident both in terms of quantity (i.e., total organic matter content) and quality (i.e., biochemical composition) of sediment organic matter. In particular, sediment protein concentration appeared to be a good descriptor of the trophic state of the benthic systems at different spatial scales. Multivariate (MDS) analysis allowed identifying areas characterised by hypertrophic, eutrophic and meso-oligotrophic conditions and to define relative threshold levels. A classification of the trophic state of coastal systems based on protein and carbohydrate concentrations is proposed.

[Epidural vs general anaesthesia]. / Anestesia epidurale vs generale.

BACKGROUND: Aim of this study is to determine if and how the anaesthesia technique can significantly influence the outcome in patients after major orthopaedic surgery in terms of: patrimony of red blood cells (blood loss and erythropoiesis), incidence of intra and postoperative complications, postoperative pain control and hospital stay. METHODS: 210 patients, ASA physical status I-III, undergoing elective primary total hip replacement were randomly allocated in three groups of 70 patients to receive either epidural anaesthesia (Group EA), general anaesthesia (GA), or epidural anaesthesia integrated with mild general anaesthesia (IA). RESULTS: Data show a significant difference between the amount of pain measured by VRS immediately after surgery: prevalently absent in groups IA (84.3%) and EA (85.7%) and prevalently severe and moderate in group AG (34.3%). The measurement of the basic circulating erythrocyte mass in the first, third and fifth postoperative day, calculated by the Mercuriali formula, which considers blood loss, autologous and homologous transfusions and erythropoiesis, showed that general anaesthesia leads to a significant delay in the resumption of haemopoiesis. This result was attenuated by its combination with epidural anaesthesia. CONCLUSIONS: On the basis of the literature and the results of our study, epidural anaesthesia seems to be the most appropriate technique for patients scheduled for total hip replacement: due to its simpler analgesic cover, its tendency to be associated with a lower incidence of complications in the first 24 hours after surgery. The incidence of relevant hypotension is minor compared to integrated anaesthesia. General anaesthesia produced a significant decrease in postoperative erythropoiesis.

Small-Scale Distribution of Bacteria, Enzymatic Activities, and Organic Matter in Coastal Sediments.

The small-scale distribution of several structural (bacterial abundance, phytopigment, total and soluble protein, and carbohydrate content) and functional ecological variables (enzymatic activities, frequency of dividing cells) was investigated in coastal sediments during a spring bloom. For bacterial abundance, enzymatic activity, and organic matter determinations, samples were collected at 5-m depth from a sediment surface delimited by a 42 x 42-cm frame, divided into 49 squares. In order to test the influence of the bottom microtopography on the investigated variables, the size of this frame was defined to cover the distance between two subsequent ripples. As indicated by Fisher's index, benthic bacteria, enzymatic activities, proteins, carbohydrates, and their soluble fractions exhibited an aggregate distribution. Sampling size (i.e., sample unit of 36 cm2) was appropriate for all variables, except for chlorophyll a and frequency of dividing cells that displayed a contagious distribution. To estimate the reliability of the current sampling strategy, we compared the mean values from three randomly selected sample units with the average value of the entire sediment surface (i.e., 49 samples). For all variables reported in this study, the use of three replicates was representative of the mean values of the sampled area with a confidence limit within +/-20%. Bacterial population sizes did not correlate with their potential food sources (e.g., phytopigments, proteins, and carbohydrates), or with enzymatic activities, suggesting the presence of possible time lags between organic inputs and microbial response. Chlorophyll a during the spring bloom displayed much higher concentrations than phaeopigments and correlated with carbohydrates. Chlorophyll a distribution was autocorrelated and displayed a large patch size (1,134 cm2). Phaeopigments and proteins accumulated in the central depression of the ripple-mark structure, apparently depending upon a passive accumulation due to the reduced current action. In contrast, microphytobenthic biomass and bacterial numbers were highest in the two opposite ripples, suggesting that different driving forces operate selectively on the living components.