The response of the algae Fucus virsoides (Fucales, Ochrophyta) to Roundup® solution exposure: A metabolomics approach.

Glyphosate, as a broad-spectrum herbicide, is frequently detected in water and several studies have investigated its effects on several freshwater aquatic organisms. Yet, only few investigations have been performed on marine macroalgae. Here, we studied both the metabolomics responses and the effect on primary production in the endemic brown algae Fucus virsoides exposed to different concentration (0, 0.5, 1.5 and 2.5 mg L-1) of a commercial glyphosate-based herbicide, namely Roundup®. Our results show that Roundup® significantly reduced quantum yield of photosynthesis (Fv/Fm) and caused alteration in the metabolomic profiles of exposed thalli compared to controls. Together with the decrease in the aromatic amino acids (phenylalanine and tyrosine), an increase in shikimate content was detected. The branched-amino acids differently varied according to levels of herbicide exposure, as well as observed for the content of choline, formate, glucose, malonate and fumarate. Our results suggest that marine primary producers could be largely affected by the agricultural land use, this asking for further studies addressing the ecosystem-level effects of glyphosate-based herbicides in coastal waters.

Tumor grade and molecular subtypes on local control in breast cancer radiotherapy: Does fractionation really matter? A retrospective control study group.

The aim of this current study was to assess whether the tumour grade and molecular subtypes have influenced local control in the whole breast hypofractionated radiotherapy (HRT) over standard radiotherapy (SRT) in early breast node negative cancer patients by a retrospective control group study. Data of 215 patients treated with hypofractionated radiotherapy at our institution from 2008 to 2011 were prospectively collected and then compared with 215 pts treated with SRT in a control group study. The local relapse free survival (LRFS) in both arms was compared on the basis of variables defined by tumour grade (Nottingham Grading System), and Molecular subtypes. Kaplan-Meier method was applied to estimate the LRFS in both groups. Chi-squared and univariate Cox proportional hazards model were conducted for all variables in both groups to assess the impact on local control. Statistical significance was assumed at P < .05. Statistical significant variables at univariate analysis were then included in multivariate Cox proportional hazards model. The median follow up duration was 9.5 years (7-13 yrs); the Kaplan Meyer 8 year LRFS did not reach any statistical significant difference between the two groups (P = . 836). At univariate Cox analysis tumour grade 3 was significantly related to local relapse only in the SRT group (P = .041) while, among molecular subtypes, no differences were found for all groups; for Her2 + noL no difference was found (P = .233). Multivariate analysis confirmed Her2 non-luminal subtype as an independent variable for local relapse regardless the fractionation arm (P = .045). Breast cancer subtypes show a different radiosensitivity, which is independent by fractionation.

Assessing the effectiveness of surrogates for species over time: Evidence from decadal monitoring of a Mediterranean transitional water ecosystem.

The use of higher taxa or alternative approach to species surrogacy, such as the BestAgg procedure, could represent cost-effective solutions to avoid expensive species-level identifications in monitoring activities, especially on the long term. However, whether a set of surrogates would be effective in subsequent reiteration of the same assessment remains largely unsolved. We used a long-term dataset on macro-benthic assemblages to test the hypothesis that family-level and BestAgg surrogates which are effective for a limited period of monitoring could be successfully applied to quantify community patterns also in subsequent monitoring programmes. The effectiveness of surrogates in detecting temporal variations in assemblage structure as at species level remained basically unaffected over a decade. Recognizing once and for all if species surrogacy may have a practical value for monitoring will strongly depend on future assessments of the potential of surrogates to reflect community changes and to retain this prerogative over time.

A regional assessment of cumulative impact mapping on Mediterranean coralligenous outcrops.

In the last decade, the 'Cumulative Pressure and Impact Assessment' (CPIA) approach emerged as a tool to map expected impacts on marine ecosystems. However, CPIA assumes a linear response of ecosystems to increasing level of cumulative pressure weighting sensitivity to different anthropogenic pressures through expert judgement. We applied CPIA to Mediterranean coralligenous outcrops over 1000 km of the Italian coastline. Extensive field surveys were conducted to assess the actual condition of coralligenous assemblages at varying levels of human pressure. As pressure increased, a clear shift from bioconstructors to turf-dominated assemblages was found. The linear model originally assumed for CPIA did not fit the actual relationship between expected cumulative impact versus assemblage degradation. A log-log model, instead, best fitted the data and predicted a different map of cumulative impact in the study area able to appreciate the whole range of impact scenarios. Hence, the relative importance of different drivers in explaining the observed pattern of degradation was not aligned with weights from the expert opinion. Such findings stress the need for more incisive efforts to collect empirical evidence on ecosystem-specific responses to human pressure in order to refine CPIA predictions.

VMAT radiation-induced nausea and vomiting in adjuvant breast cancer radiotherapy: The incidental effect of low-dose bath exposure.

BACKGROUND AND PURPOSE: To investigate the hypothesis on low-dose bath exposure related to radiation-induced nausea and vomiting (RINV) in adjuvant breast volumetric modulated arch therapy (VMAT). METHODS AND MATERIALS: A total of 106 consecutive breast cancer patients (pts) treated with adjuvant radiotherapy (RT) with VMAT from January 2013 to May 2016 were evaluated retrospectively. For each pt, a planning CT was reimported and the coeliac plexus and gastroesophageal junction with gastric mouth (GEJCPs) were contoured as a new organ at risk (OAR) in the upper abdominal area. RINV was associated with Dmax and Dmean to GEJCPs. Univariate analysis with χ2, t-test, and Pearson's covariance was used for statistical analysis. RESULTS: Of 106 pts, 64% complained of acute RINV according to the Common Terminology Criteria for Adverse Events (CTCAE) version 4.03. RINV was related to Dmax > 10 Gy and Dmean > 3 Gy to GEJCPs (P < 0.005). The radiation breast side and planning target volume (PTV) correlated with RINV. CONCLUSIONS: RINV in VMAT breast radiotherapy could be a new emerging acute side effect due to a low dose bath to upper abdominal structures such as the GEJCPs. A Dmax < 10 Gy and Dmean < 3 Gy to GEJCPs should be constrained in VMAT planning to minimize RINV risk in breast radiotherapy.

Impact of offshore gas platforms on the structural and functional biodiversity of nematodes.

The Mediterranean Sea hosts hundreds of offshore gas platforms, whose activity represents a potential threat to marine ecosystems. Evidence from several studies indicates that nematodes can be highly sensitive to changes in the environmental quality. Here, we investigated the response of nematode assemblages to the presence of offshore gas platforms (located in the central Mediterranean Sea) in terms of spatial heterogeneity, structural and functional diversity. Since the effect of the investigated offshore platforms on macrofaunal assemblages were previously assessed by Terlizzi et al. (2008), the study provided also the opportunity to compare the response of different benthic compartments to the same impact related to fossil fuel extraction on marine environments. The platforms had a significant impact on nematode assemblages up to 1000 m distance from the structure. The effects were evident in term of: a) more homogeneous spatial distribution of nematode assemblages, b) increased trophic diversity of deposit feeders and c) changes in life strategies with an increase of opportunistic species in sediments closer to the platforms. Such effects seemed to be related to the dimension of the platform structures, rather than to chemical pollution or changes in food availability. These findings suggest that the platforms exert a physical alteration of the surrounding environment that is reflected by altered structural and functional traits of nematode biodiversity. The use of nematodes for monitoring the effects of the platforms only partially matched with the results obtained using macrofauna, providing further insights on potential outcomes on the functional response of marine assemblages to fossil fuel extraction.

Could molecular effects of Caulerpa racemosa metabolites modulate the impact on fish populations of Diplodus sargus?

The green alga Caulerpa racemosa is a non-native, invasive species in the Mediterranean, and an important stressor for several native organisms. The algal capacity to produce secondary metabolites has been suggested to modulate success of the C. racemosa invasion, although many of potentially involved biological pathways still remain unexplored. In this respect, the aim of the present study was to investigate some molecular and cellular effects in the white seabream Diplodus sargus, an ecologically key species, which included the alien C. racemosa in its diet. Organisms were sampled in 2 seasonal periods from 3 locations of Southern Italy, each characterized by different levels of algal abundance. The level of caulerpin, one of the main secondary algal metabolites, in fish tissues has been used as an indicator of the trophic exposure to the seaweed and related with molecular and cellular responses. Chemical analyses indicated that fish from invaded sites can accumulate caulerpin, with liver concentrations ranging from a few up to hundreds of µg/g. Biomarkers analyses revealed only limited alterations of the main antioxidant defences, such as glutathione reductase and levels of glutathione; on the other hand, increased enzymatic activities of cytochrome P450, glutathione S-transferases and acyl CoA oxidase, as well as enhanced gene transcription for peroxisome proliferator-activated receptor alpha, cytochrome P4501A and vitellogenin 1 were observed in fish more exposed to C. racemosa as indicated by liver concentrations of caulerpin higher than 50 µg/g. Despite a direct molecular relationship with this algal metabolite could not be established, our results suggest that a C. racemosa enriched diet can modulate biotransformation and fatty acids metabolism of D. sargus. Assessing whether similar effects represent short- or long-term effects will be of crucial importance to understand consequences on the general health status and reproductive performance of exposed key fish species in the Mediterranean region.

Human trampling as short-term disturbance on intertidal mudflats: effects on macrofauna biodiversity and population dynamics of bivalves.

The effect of physical disturbance in the form of trampling on the benthic environment of an intertidal mudflat was investigated. Intense trampling was created as unintended side-effect by benthic ecologists during field experiments in spring and summer 2005, when a mid-shore area of 25 × 25 m was visited twice per month by on average five researchers for a period of 8 months. At the putatively-impacted location (I) (25 × 25 m) and two nearby control locations (Cs) (25 × 25 m each), three sites (4 × 4 m) were randomly selected and at each site, three plots (50 × 50 cm) were sampled after 18 and 40 days from the end of the disturbance. Multivariate and univariate asymmetrical analyses tested for changes in the macrofaunal assemblage, biomass of microphytobenthos and various sediment properties (grain-size, water content, NH4 and NO3 concentrations in the pore water) between the two control locations (Cs) and the putatively-impacted location (I). There were no detectable changes in the sediment properties and microphytobenthos biomass, but variability at small scale was observed. Microphytobenthos and NH4 were correlated at I to the number of footprints, as estimated by the percentage cover of physical depressions. This indicated that trampling could have an impact at small scales, but more investigation is needed. Trampling, instead, clearly modified the abundance and population dynamics of the clam Macoma balthica (L.) and the cockle Cerastoderma edule (L.). There was a negative impact on adults of both species, probably because footsteps directly killed or buried the animals, provoking asphyxia. Conversely, trampling indirectly enhanced recruitment rate of M. balthica, while small-sized C. edule did not react to the trampling. It was likely that small animals could recover more quickly because trampling occurred during the growing season and there was a continuous supply of larvae and juveniles. In addition, trampling might have weakened negative adult-juvenile interactions between adult cockles and juvenile M. balthica, thus facilitating the recruitment. Our findings indicated that human trampling is a relevant source of disturbance for the conservation and management of mudflats. During the growing season recovery can be fast, but in the long-term it might lead towards the dominance of M. balthica to the cost of C. edule, thereby affecting ecosystem functioning.

Structural and functional response of meiofauna rocky assemblages to sewage pollution.

Despite meiofauna being one of the most popular tool for detecting the ecological effects of different sources of impact, the application of structured experimental designs to this benthic component is typically neglected, especially in subtidal rocky shores. In this study, an asymmetrical design was used to detect hard-bottom meiofaunal abundance and biodiversity response to sewage discharge. Sampling was carried out at a depth of 3-4 m by SCUBA diving, by means of a modified manual corer. Six replicate cores were collected at three sites (80-100 m apart), at each of three locations (one purportedly impacted [I] and two controls [Cs]). A total of 84,994 specimens were collected. The outfall affected meiofaunal assemblages in terms of taxon richness, by determining a significant reduction of the number of taxa in the disturbed location, and community structure, by causing functional changes in terms of a decrease of the abundance of nematodes and an increase of the hydrozoan component. Nematodes and syllid polychaetes showed significant lower average abundance at I. Multivariate analyses showed that both meiofaunal assemblage and syllids were significantly different at I compared with Cs. The sewage outfall also affected patterns of spatial distribution at the scale of site (100 m apart) and of replicate units (centimetres apart), both in syllids and in nematodes. Our results provide evidence that the selection of multiple controls is crucial to prevent the widespread risk of Type II error, highlighting the need of more accurate experimental designs when dealing with meiofauna.

Taxonomic sufficiency and the increasing insufficiency of taxonomic expertise.

Taxonomic sufficiency (TS) involves the identification of taxa only to a level of taxonomic resolution sufficient to permit the detection of changes in stressed assemblages. Recently, however, TS has been proposed also for conservation issues as a tool to estimate biodiversity over large areas and in poorly known environments. This paper briefly reviews the use of TS in environmental impact studies and the effects of TS on sampling procedures and data analyses. The risk of possible loss of information depending on TS and the studied environment are discussed. Concluding remarks deal with the dangers of loss of taxonomic expertise in marine biological studies and assess critically the proposal of TS as a tool to describe biodiversity at a taxonomic level higher than species.

The effects of sewage discharge on shallow hard substrate sessile assemblages.

On rocky shores, sewage discharges can modify natural distribution patterns of sessile organisms. The impact of sewage on shallow hard substrate assemblages has been assessed along SW Apulian coast (Ionian Sea, Italy), providing a framework to evaluate the benefits of future sewage displacement to deeper waters. Four locations (three controls and one putatively impacted) were selected and three sites were chosen at each location. Each site was sampled by 10 replicate photographic records. Univariate analyses revealed that the outfall did not affect the spatial distribution of number of taxa, total cover and abundance of some dominant taxa (mostly algae, sponges and bryozoans). The outfall negatively influenced the natural distribution pattern of filamentous green algae, whilst some algae (i.e. Gelidiales and Colpomenia sinuosa) were exclusively present at the impacted location. Multivariate analyses revealed that the outfall heavily modified the natural pattern of variability in the structure of the assemblage.

Coastal fish indicate human-induced changes in the Mediterranean littoral.

Coastal fish assemblages were studied to assess two sorts of human impacts in southwestern Apulia (SE Italy, Mediterranean Sea). Fish assemblages were evaluated by visual census along two rocky locations impacted by a sewage outfall discharging nearshore (S) and by date-mussel (Lithophaga lithophaga) fisheries (F), respectively, and at two control locations (Cs). Multivariate analyses showed that fish assemblage structures at S and F differed from those at Cs. Asymmetric ANOVAs indicated that species richness were significantly lower both at S (approximately 27%) and at F (approximately 35%) compared with Cs. Total fish abundance was 5- to 7-fold higher at S than at Cs, while the values recorded at F were comparable to those of Cs. At S, average abundances of planktivorous fish and POM feeders were higher, and those of labrids and sparids of the genus Diplodus were lower, respectively, than at Cs. Labrids of the genus Symphodus and small serranids were significantly less abundant at F than Cs. Data suggested that coastal fish respond to the impact caused by the sewage discharge and provided a framework to assess potential benefits of its future displacement to deeper waters. For the first time, moreover, this study provided suggestive evidence that the habitat destruction caused by the illegal date-mussel fisheries may affect fish assemblages.