Dietary Salicornia ramosissima improves the European seabass (Dicentrarchus labrax) inflammatory response against Photobacterium damselae piscicida.

Introduction: Modern fish farming faces challenges in sourcing feed ingredients, most related with their prices, 21 availability, and specifically for plant protein sources, competition for the limited cultivation space for 22 vegetable crops. In that sense, halophytes have the added value of being rich in valuable bioactive compounds and salt tolerant. This study assessed the inclusion of non-food fractions of S. ramosissima in European seabass diets. Methods: Different levels (2.5%, 5%, and 10%) were incorporated into seabass diets, replacing wheat meal (diets ST2.5, ST5, and ST10) or without inclusion (CTRL). Experimental diets were administered to seabass juveniles (8.62 ± 0.63 g) for 34 and 62 days and subsequent inflammatory responses to a heat-inactivated Photobacterium damselae subsp. piscicida (Phdp) were evaluated in a time-course manner (4, 24, 48, and 72 h after the challenge). At each sampling point, seabass haematological profile, plasma immune parameters, and head-kidney immune-related gene expression were evaluated. Results: After both feeding periods, most parameters remained unaltered by S. ramosissima inclusion; nonetheless, seabass fed ST10 showed an upregulation of macrophage colony-stimulating factor 1 receptor 1 (mcsf1r1) and cluster of differentiation 8 (cd8ß) compared with those fed CTRL after 62 days of feeding. Regarding the inflammatory response, seabass fed ST10 showed lower plasma lysozyme levels than their counterparts fed ST2.5 and ST5 at 24 h following injection, while 4 h after the inflammatory stimulus, seabass fed ST10 presented higher numbers of peritoneal leucocytes than fish fed CTRL. Moreover, at 4 h, fish fed ST2.5, ST5, and ST10 showed a higher expression of interleukin 1ß (il1ß), while fish fed ST5 showed higher levels of ornithine decarboxylase (odc) than those fed CTRL. An upregulation of macrophage colony-stimulating factor 1 receptor 1 (mcsf1r1) and glutathione peroxidase (gpx) was also observed at 72 h in fish fed ST10 or ST5 and ST10 compared with CTRL, respectively. Discussion: In conclusion, incorporating up to 10% of the non-food fraction S. ramosissima in feed did not compromise seabass growth or immune status after 62 days, aligning with circular economy principles. However, S. ramosissima inclusion improved the leucocyte response and upregulated key immune-related genes in seabass challenged with an inactivated pathogen.

Salicornia ramosissima Biomass as a Partial Replacement of Wheat Meal in Diets for Juvenile European Seabass (Dicentrarchus labrax).

The green tips of Salicornia ramosissima are used for human consumption, while, in a production scenario, the rest of the plant is considered a residue. This study evaluated the potential of incorporating salicornia by-products in diets for juvenile European seabass, partially replacing wheat meal, aspiring to contribute to their valorization. A standard diet and three experimental diets including salicornia in 2.5%, 5% and 10% inclusion levels were tested in triplicate. After 62 days of feeding, no significant differences between treatments were observed in fish growth performances, feeding efficiency and economic conversation ratio. Nutrient digestibility of the experimental diets was unaffected by the inclusion of salicornia when compared to a standard diet. Additionally, salicornia had significant modulatory effects on the fish muscle biochemical profiles, namely by significantly decreasing lactic acid and increasing succinic acid levels, which can potentially signal health-promoting effects for the fish. Increases in DHA levels in fish fed a diet containing 10% salicornia were also shown. Therefore, the results suggest that salicornia by-products are a viable alternative to partially replace wheat meal in diets for juvenile European seabass, contributing to the valorization of a residue and the implementation of a circular economy paradigm in halophyte farming and aquaculture.

Impacts of humic substances, elevated temperature, and UVB radiation on bacterial communities of the marine sponge Chondrilla sp.

Sponges are abundant components of coral reefs known for their filtration capabilities and intricate interactions with microbes. They play a crucial role in maintaining the ecological balance of coral reefs. Humic substances (HS) affect bacterial communities across terrestrial, freshwater, and marine ecosystems. However, the specific effects of HS on sponge-associated microbial symbionts have largely been neglected. Here, we used a randomized-controlled microcosm setup to investigate the independent and interactive effects of HS, elevated temperature, and UVB radiation on bacterial communities associated with the sponge Chondrilla sp. Our results indicated the presence of a core bacterial community consisting of relatively abundant members, apparently resilient to the tested environmental perturbations, alongside a variable bacterial community. Elevated temperature positively affected the relative abundances of ASVs related to Planctomycetales and members of the families Pseudohongiellaceae and Hyphomonadaceae. HS increased the relative abundances of several ASVs potentially involved in recalcitrant organic matter degradation (e.g., the BD2-11 terrestrial group, Saccharimonadales, and SAR202 clade). There was no significant independent effect of UVB and there were no significant interactive effects of HS, heat, and UVB on bacterial diversity and composition. The significant, independent impact of HS on the composition of sponge bacterial communities suggests that alterations to HS inputs may have cascading effects on adjacent marine ecosystems.

Facemasks: An insight into their abundance in wetlands, degradation, and potential ecotoxicity.

Disposable facemasks represent a new form of environmental contamination worldwide. This study aimed at addressing the abundance of facemasks in an overlooked natural environment with high ecological and economic value - the wetlands (Ria de Aveiro, Portugal, as study case), evaluating their potential biodegradation using naturally occurring fungi and assessing the potential ecotoxicity of released microfibres on local bivalves. All masks collected within 6500 m2 area of Aveiro wetland were 100 % disposable ones (PP-based, confirmed by Fourier transform infrared spectroscopy - FTIR) with an initial abundance of 0.0023 items/m2 in Sept. 2021, which was reduced by ∼40 % in Apr. 2022 and ∼87 % in Sept. 2022, as a reflection of the government policies. Analysis of the carbonyl index (0.03 to 1.79) underlined their state of degradation, primarily due to sun exposure during low tides. In laboratory conditions, 1 mm2 microplastics obtained from new disposable facemasks were prone to biodegradation by Penicillium brevicompactum and Zalerion maritimum inferred from microplastics mass loss (∼22 to -26 % and ∼40 to 50 %, respectively) and FTIR spectra (particularly in the hydroxyl and carbonyl groups). In addition, microfibres released from facemasks induced sublethal effects on the clam, Venerupis corrugata, mostly in their UV-aged form when compared to pristine ones, characterised by a decrease in cellular energy allocation (CEA) and an increase in aerobic energy metabolism (ETS). Concomitantly, clams exposed to 1250 items/L of UV-aged microplastics (similar to field-reported concentrations) expressed greater clearance capacity, indicating a need to compensate for the potential energy unbalance. This study provides the first baseline monitoring of facemasks in wetlands while bringing new evidence on their biodegradation and ecotoxicity, considering environmentally relevant conditions and keystone organisms in such environments. Such studies require scientific attention for rapid regulatory action against this emerging and persistent pollutant, also targeting remediation and mitigation strategies considering these items under pandemic scenarios.

Microencapsulated Diets as an Alternative to Bivalve Feeding: Particle Size and Microalga Content Affect Feed Intake.

Bivalve mollusks represent a nutritious source with a low environmental impact; as a result, they are one of the most attractive aquaculture options. Advances in microencapsulation technology offer great potential to face key bivalve nutrition problems, and an alga-based microencapsulated diet can turn enriched bivalves into potential functional foods. The central goal of this study was the evaluation of food intake as a function of particle size and microalga content following the supply of four microencapsulated diets, incorporating as core material Nannochloropsis sp. or Tetraselmis sp. in 20 or 40 µm diameter pellets (diets N20, T20, N40, and T40, respectively) in five bivalve species (Magallana gigas, Solen marginatus, Ruditapes decussatus, Ruditapes philippinarum, and Cerastoderma edule). Overall, all tested diets were easily ingested, although food intake was higher for N20 (except for the S. marginatus, which showed a higher rate for the diet T40). Concerning a size-related analysis, C. edule and S. marginatus favored, respectively, smaller and bigger pellet-sized diets, with no signs of selectivity for microalga species. The diet T20 was the lesser ingested, except for C. edule. This knowledge enables a better selection of feed with appropriate and species-adjusted profiles, contributing to the optimization of microencapsulated diets for bivalve rearing and a better final product.

Health-Promoting Additives Supplemented in Inert Microdiets for Whiteleg Shrimp (Penaeus vannamei) Post-Larvae: Effects on Growth, Survival, and Health Status.

Dietary additives have the potential to stimulate the whiteleg shrimp immune system, but information is scarce on their use in diets for larval/post-larval stages. The potential beneficial effects of vitamins C and E, ß-glucans, taurine, and methionine were evaluated. Four experimental microdiets were tested: a positive control diet (PC); the PC with decreased levels of vitamin C and E as negative control (NC); the PC with increased taurine and methionine levels (T + M); and the PC supplemented with ß-glucans (BG). No changes in growth performance and survival were observed. However, post-larvae shrimp fed the NC had lower relative expressions of pen-3 than those fed the PC, suggesting that lower levels of vitamins C and E may impact the shrimp immune status. Lipid peroxidation levels dropped significantly in the BG compared to the PC, indicating that ß-glucans improved the post-larvae antioxidant mechanisms. Furthermore, when compared with the NC diet, PL fed with BG showed significant increases in tGSH levels and in the relative expression of crus and pen-3, suggesting a synergistic effect between vitamins C and E and ß-glucans. Amongst the additives tested, ß-glucans seems to be the most promising even when compared to a high-quality control diet.

Exploring the Antioxidant and Genoprotective Potential of Salicornia ramosissima Incorporation in the Diet of the European Seabass (Dicentrarchus labrax).

The identification of novel feed materials as a source of functional ingredients is a topical priority in the finfish aquaculture sector. Due to the agrotechnical practices associated and phytochemical profiling, halophytes emerge as a new source of feedstuff for aquafeeds, with the potential to boost productivity and environmental sustainability. Therefore, the present study aimed to assess the potential of Salicornia ramosissima incorporation (2.5, 5, and 10%), for 2 months, in the diet of juvenile European seabass, seeking antioxidant (in the liver, gills, and blood) and genoprotective (DNA and chromosomal integrity in blood) benefits. Halophyte inclusion showed no impairments on growth performance. Moreover, a tissue-specific antioxidant improvement was apparent, namely through the GSH-related defense subsystem, but revealing multiple and complex mechanisms. A genotoxic trigger (regarded as a pro-genoprotective mechanism) was identified in the first month of supplementation. A clear protection of DNA integrity was detected in the second month, for all the supplementation levels (and the most prominent melioration at 10%). Overall, these results pointed out a functionality of S. ramosissima-supplemented diets and a promising way to improve aquaculture practices, also unraveling a complementary novel, low-value raw material, and a path to its valorization.

Can the toxicity of polyethylene microplastics and engineered nanoclays on flatfish (Solea senegalensis) be influenced by the presence of each other?

Microplastics and nanomaterials are applied in a myriad of commercial and industrial applications. When leaked to natural environments, such small particles might threaten living organisms' health, particularly when considering their potential combination that remains poorly investigated. This study investigated the physiological and biochemical effects of polyethylene (PE; 64-125 µm in size, 0.1, 1.0, and 10.0 mg·L-1) single and combined with an engineered nanomaterial applied in antifouling coatings, the copper-aluminium layered double hydroxides (Cu-Al LDH; 0.33, 1.0, and 3.33 mg·L-1) in the flatfish Solea senegalensis larvae (8 dph) after 3 h exposure, in a full factorial design. Particles ingestion, histopathology, and biochemical biomarkers were assessed. Fish larvae presented <1 PE particles in their gut, independently of their concentration in the medium. The histological health index showed minimal pathological alterations at PE combined exposure, with a higher value observed at 1 mg LDH·L-1 × 0.1 mg PE·L-1. Gut deformity and increased antioxidant defences (catalase), neurotransmission (acetylcholinesterase), and aerobic energy production (electron transport system) were observed at PE ≥ 1.0 mg·L-1. No oxidative damage (lipid peroxidation) or alterations in the detoxification capacity (glutathione-S-transferase) was observed on single and combined exposures. PE, combined or not with Cu-Al LDH, does not seem to compromise larvae's homeostasis considering levels reported so far in the marine and aquaculture environments. However, harsh effects are expected with MP contamination rise, as projections suggest.

Dietary Natural Plant Extracts Can Promote Growth and Modulate Oxidative Status of Senegalese Sole Postlarvae under Standard/Challenge Conditions.

Plant extracts are known for their high content and diversity of polyphenols, which can improve fish oxidative status. A growth trial with Senegalese sole postlarvae (45 days after hatching) fed with one of four experimental diets-control (CTRL), and supplemented with curcumin (CC), green tea (GT), and grape seed (GS) extracts-was performed to assess if supplementation could improve growth performance and oxidative status. At the end of the growth trial, postlarvae were submitted to a thermal stress to assess their robustness. Sole growth was improved by CC and GS diets when compared to those fed the CTRL. CC and CTRL postlarvae presented the lowest oxidative damage (lipid peroxidation and protein carbonylation values). Stress-related biomarkers (heat shock protein 70 and glutathione-S-transferase) decreased in CC fish compared to those fed the CTRL diet, which might be due to a direct antioxidant capacity. In contrast, oxidative damage increased in GT and GS sole reared in standard conditions. However, after a thermal stress, GT and GS diets prevented the increase of protein carbonylation content and the decrease of antioxidant glutathione, depending on exposure time. Overall, dietary supplementation with natural extracts modulated oxidative status and stress response after a short/long-term exposure to temperature.

Ocean Warming May Enhance Biochemical Alterations Induced by an Invasive Seaweed Exudate in the Mussel Mytilus galloprovincialis.

Ocean warming and biological invasions are among the most pervasive factors threatening coastal ecosystems with a potential to interact. Ongoing temperature rise may affect physiological and cellular mechanisms in marine organisms. Moreover, non-indigenous species spread has been a major challenge to biodiversity and ecosystem functions and services. The invasive red seaweed Asparagopsis armata has become successfully established in Europe. Its exudate has been considered deleterious to surrounding native species, but no information exists on its effect under forecasted temperature increase. This study evaluated the combined effects of temperature rise and A. armata exudate exposure on the native mussel Mytilus galloprovincialis. Oxidative stress, neurophysiological and metabolism related biomarkers were evaluated after a 96 h-exposure to exudate (0% and 2%) under present (20 °C) and warming (24 °C) temperature scenarios. Short-term exposure to A. armata exudate affected the oxidative stress status and neurophysiology of the mussels, with a tendency to an increasing toxic action under warming. Significant oxidative damage at protein level was observed in the digestive gland and muscle of individuals exposed simultaneously to the exudate and temperature rise. Thus, under a climate change scenario, it may be expected that prolonged exposure to the combined action of both stressors may compromise M. galloprovincialis fitness and survival.

Organic solvents alter photophysiological and oxidative stress profiles of the coral Zoanthus sp. - Towards an optimization of ecotoxicological protocols.

Coral reefs are declining, affected by climate change and escalating anthropogenic pressures, such as pollution or habitat alteration. Consequently, ecotoxicological assays with tropical corals have increased, specifically towards the study of emergent or persistent pollutants. However, standardized methodology to test for corals is non-existent, and their response to organic solvents, recurrently required in ecotoxicological appraisals, remains unknown. Therefore, we aimed to establish a threshold for the safe use of the selected solvents in ecotoxicological studies with these organisms. We assessed the oxidative stress response (antioxidant response and oxidative damage), cellular energy allocation and photophysiology of the photosynthetic coral Zoanthus sp. (Anthozoa, Hexacorallia) exposed to six doses of three different organic solvents (ethanol, methanol and dimethyl sulfoxide - DMSO). Our results suggest that the coral is more sensitive to methanol and DMSO than to ethanol. Methanol and DMSO LOEC were 0.01 mL L-1 affecting maximum quantum yield (Fv/Fm) and glutathione S-transferase (GST) activity, respectively, while for ethanol was 0.03 mL L-1, influencing Fv/Fm. Despite the higher tolerance of Zoanthus sp. to ethanol, 2.9 mL L-1 of this organic solvent was the only treatment causing mortality. Based on these findings, thresholds for the use of organic solvents with tropical corals can now be adopted. Nevertheless, species specificities should not be overlooked.

Vulnerability of Tritia reticulata (L.) early life stages to ocean acidification and warming.

Ocean acidification and warming (OA-W) result mainly from the absorption of carbon dioxide and heat by the oceans, altering its physical and chemical properties and affecting carbonate secretion by marine calcifiers such as gastropods. These processes are ongoing, and the projections of their aggravation are not encouraging. This work assesses the concomitant effect of the predicted pH decrease and temperature rise on early life stages of the neogastropod Tritia reticulata (L.), a common scavenger of high ecological importance on coastal ecosystems of the NE Atlantic. Veligers were exposed for 14 days to 12 OA-W experimental scenarios generated by a factorial design of three pH levels (targeting 8.1, 7.8 and 7.5) at four temperatures (16, 18, 20 and 22 °C). Results reveal effects of both pH and temperature (T °C) on larval development, growth, shell integrity and survival, individually or interactively at different exposure times. All endpoints were initially driven by pH, with impaired development and high mortalities being recorded in the first week, constrained by the most acidic scenarios (pHtarget 7.5). Development was also significantly driven by T °C, and its acceleration with warming was observed for the remaining exposure time. Still, by the end of this 2-weeks trial, larval performance and survival were highly affected by the interaction between pH and T °C: growth under warming was evident but only for T °C ≤ 20 °C and carbonate saturation (pHtarget ≥ 7.8). In fact, carbonate undersaturation rendered critical larval mortality (100%) at 22 °C, and the occurrence of extremely vulnerable, unshelled specimens in all other tested temperatures. As recruitment cohorts are the foundation for future populations, our results point towards the extreme vulnerability of this species in case tested scenarios become effective that, according to the IPCC, are projected for the northern hemisphere, where this species is ubiquitous, by the end of the century. Increased veliger mortality associated with reduced growth rates, shell dissolution and loss under OA-W projected scenarios will reduce larval performance, jeopardizing T. reticulata subsistence.

The impacts of warming on the toxicity of carbon nanotubes in mussels.

With the increased production and research on nanoparticles, the presence of carbon nanotubes (CNTs) in aquatic systems is very likely to increase. Although it has been shown that CNTs may cause toxicity in marine organisms, to our knowledge, the possible impacts under global temperature increase is still unknown. For this reason, biochemical and physiological impacts induced in Mytilus galloprovincialis due to the presence of functionalized multi-walled CNTs (f-MWCNTs) and increased temperature were investigated in the present study. The mussels exposed to increased temperature alone presented higher metabolic capacity and expenditure of glycogen as an energy resource to fuel up defense mechanisms and thus preventing oxidative damage. Contrarily, organisms exposed to f-MWCNTs alone seemed not stressed enough to demonstrate differences in the metabolism capacity. Furthermore, f-MWCNTs seemed not able to significantly activate their antioxidant and biotransformation enzymes, which in turn may led to oxidative damage in the cells especially when organisms were exposed to a warmer temperature. In fact, at higher temperature, the antioxidant response of organisms exposed to f-MWCNTs was not effective and oxidative damage levels were observed. Nevertheless, no additive or synergetic effects were observed when mussels were exposed to both stressors simultaneously.

Are the effects induced by increased temperature enhanced in Mytilus galloprovincialis submitted to air exposure?

Intertidal mussel species are frequently exposed to changes of environmental parameters related to tidal regimes that include a multitude of stressors that they must avoid or tolerate by developing adaptive strategies. In particular, besides air exposure during low tides, intertidal mussels are also subjected to warming and, consequently, to higher risk of desiccation. However, scarce information is available regarding the responses of mussels to tidal regimes, particularly in the presence of other stressors such as increased temperature. Investigating the impacts of such combination of conditions will allow to understand the possible impacts that both factors interaction may generate to these intertidal organisms. To this end, the present study evaluated the impacts of different temperatures (18 °C and 21 °C) on Mytilus galloprovincialis when continuously submersed or exposed to a tidal regime for 14 days. Results showed that in mussels exposed to increased temperature under submersion conditions, the stress induced was enough to activate mussels' antioxidant defenses (namely glutathione peroxidase, GPx), preventing oxidative damage (lipid peroxidation, LPO; protein carbonylation, PC). In mussels exposed to tides at control temperature, metabolic capacity increased (electron transport system activity, ETS), and GPx was induced, despite resulting in increased LPO levels. Moreover, the combination of tides and temperature increase led to a significant decrease of lipid (LIP) content, activation of antioxidant defenses (superoxide dismutase, SOD; GPx) and increase of oxidized glutathione (GSSG), despite these mechanisms were not sufficient to prevent increased cellular damage. Therefore, the combination of increased temperature and air exposure induced higher oxidative stress in mussels. These findings indicate that increasing global warming could be more impacting to intertidal organisms compared to organisms continuously submersed. Furthermore, our results indicate that air exposure can act as a confounding factor when assessing the impacts of different stressors in organisms living in coastal systems.

Are the impacts of carbon nanotubes enhanced in Mytilus galloprovincialis submitted to air exposure?

Intertidal species are frequently exposed to environmental changes associated with multiple stressors, which they must either avoid or tolerate by developing physiological and biochemical strategies. Some of the natural environmental changes are related with the tidal cycle which forces organisms to tolerate the differences between an aquatic and an aerial environment. Furthermore, in these environments, organisms are also subjected to pollutants from anthropogenic sources. The present study evaluated the impacts in Mytilus galloprovincialis exposed to multi-walled carbon nanotubes (0.01 mg/L MWCNTs) when continuously submersed or exposed to tides (5 h of low tide, 7 h of high tide) for 14 days. Our results demonstrated that mussels were physiologically and biochemically affected by MWCNTs, especially when exposed to tides. In fact, when only exposed to the carbon nanoparticles or only exposed to tides, the stress induced was not enough to activate mussels' antioxidant defenses which resulted in oxidative damage. However, when mussels were exposed to the combination of tides and MWCNTs increased metabolism was observed, associated with a possible higher production of reactive oxygen species (ROS), leading to a significant increase in the activities of antioxidant enzymes (superoxide dismutase, SOD and glutathione peroxide, GPx) and oxidized glutathione content (GSSG), preventing the occurrence of cellular damage, expressed as no lipid peroxidation (LPO) or protein carbonylation (PC). Therefore, organisms seemed to be able to tolerate MWCNTs and air exposure during tidal regime; however, the combination of both stressors induced higher oxidative stress. These findings indicate that the increasing presence of carbon nanoparticles in marine ecosystems can induce higher toxic impacts in intertidal organisms compared to organisms continuously submerged. Also, our results may indicate that air exposure can act as a cofounding factor on the assessment of different stressors in organisms living in coastal systems.

Invasive Species Mediate Insecticide Effects on Community and Ecosystem Functioning.

Anthropogenic activities increase pesticide contamination and biological invasions in freshwater ecosystems. Understanding their combined effects on community structure and on ecosystem functioning presents challenges for an improved ecological risk assessment. This study focuses on an artificial stream mesocosms experiment testing for direct and indirect effects of insecticide (chlorantraniliprole - CAP) exposure on the structure of a benthic macroinvertebrate freshwater community and on ecosystem functioning (leaf decomposition, primary production). To understand how predator identity and resource quality alter the community responses to chemical stress, the mediating effects of an invasive predator species (crayfish Procambarus clarkii) and detritus quality (tested by using leaves of the invasive Eucalyptus globulus) on insecticide toxicity were also investigated. Low concentrations of CAP reduced the abundance of shredders and grazers, decreasing leaf decomposition and increasing primary production. Replacement of autochthonous predators and leaf litter by invasive species decreased macroinvertebrate survival, reduced leaf decomposition, and enhanced primary production. Structural equation modeling (SEM) highlighted that CAP toxicity to macroinvertebrates was mediated by the presence of crayfish or eucalypt leaf litter which are now common in many Mediterranean freshwaters. In summary, our results demonstrate that the presence of these two invasive species alters the effects of insecticide exposure on benthic freshwater communities. The approach used here also allowed for a mechanistic evaluation of indirect effects of these stressors and of their interaction on ecosystem functional endpoint, emphasizing the value of incorporating biotic stressors in ecotoxicological experiments.

Effect of tidal environment on the trophic balance of mixotrophic hexacorals using biochemical profile and photochemical performance as indicators.

Fluctuations of environmental factors in intertidal habitats can disrupt the trophic balance of mixotrophic cnidarians. We investigated the effect of tidal environments (subtidal, tidal pools and emerged areas) on fatty acid (FA) content of Zoanthus sociatus and Siderastrea stellata. Effect on photophysiology was also accessed as an autotrophy proxy. There was a general tendency of a lower percentage of zooplankton-associated FAs in colonies from emerged areas or tidal pools when compared with colonies from the subtidal environment. Moreover, tidal environment significantly affected the photophysiology of both species. Colonies from the subtidal generally showed lower values of α, ETRmax and Ek when compared with their conspecifics from tidal pools or emerged areas. However, the absence of consistent patterns in Fv/Fm and in dinoflagellate-associated FAs, suggest that these corals are well adapted to intertidal conditions. This suggests that intertidal pressures may disturb the trophic balance, mainly by affecting heterotrophy of these species.

Trophic and stoichiometric consequences of nutrification for the intertidal tropical zoanthid Zoanthus sociatus.

Zoanthids are conspicuous and abundant members of intertidal environments, where they are exposed to large environmental fluctuations and subject to increasing loads of anthropogenic nutrients. Here we assess the trophic ecology and stoichiometric consequences of nutrient loading for symbiotic zoanthids inhabiting different intertidal habitats. More specifically, we analysed the stable isotope signature (δ13C and δ15N), elemental composition (C, N and P) and stoichiometry (C:N, C:P, N:P) of Zoanthus sociatus differently exposed to nutrification. Results suggest that autotrophy is the main feeding mode of zoanthids and that the effect water nutrient content differently affects the elemental phenotype of zoanthids depending on tidal habitat. Additionally, habitat effects on Z. sociatus P-related stoichiometric traits highlight functional differences likely associated with variation in Symbiodinium density. These findings provide an innovative approach to assess how cnidarian-dinoflagellate symbioses response to ecosystem changes in environmentally dynamic reef flats, particularly nutrient loading.

Application of phage therapy during bivalve depuration improves Escherichia coli decontamination.

The present study investigated the potential application of the bacteriophage (or phage) phT4A, ECA2 and the phage cocktail phT4A/ECA2 to decrease the concentration of Escherichia coli during the depuration of natural and artificially contaminated cockles. Depuration in static seawater at multiplicity of infection (MOI) of 1 with single phage suspensions of phT4A and ECA2 was the best condition, as it decreased by ∼2.0 log CFU/g the concentration of E. coli in artificially contaminated cockles after a 4 h of treatment. When naturally contaminated cockles were treated in static seawater with single phage suspensions and the phage cocktail, similar decreases in the concentration of E. coli (∼0.7 log CFU/g) were achieved. However, when employing the phage cocktail, a longer treatment time was required to obtain comparable results to those achieved when using single phage suspensions. When naturally contaminated cockles were depurated with phage phT4A in a recirculated seawater system (mimicking industrial depuration conditions), a 0.6 log CFU/g reduction of E. coli was achieved after a 2 h of treatment. When the depuration process was performed without phage addition, a 4 h treatment was necessary to obtain a similar decrease. By combining phage therapy and depuration procedures, a reduction in bivalves depuration period can be achieved for, thus decreasing the cost associated with this procedure and even enhance the quality and safety of depurated bivalves destined for human consumption.

Application of bacteriophages during depuration reduces the load of Salmonella Typhimurium in cockles.

As bivalve molluscs are filter feeder, often consumed raw or lightly cooked and are frequently cultivated in contaminated waters, they are implicated in food-borne disease transmission to human. The present study investigated the potential application of bacteriophage (or phage) phSE-2, phage phSE-5 and phage cocktail phSE-2/phSE-5 to decrease the concentration of Salmonella enterica serovar Typhimurium (Salmonella Typhimurium) during the depuration of natural and artificially contaminated cockles (Cerastoderma edule). Cockles were artificially infected with 105 and 106colony-forming units (CFU)/mL of S. Typhimurium in static seawater and infected group were treated with phages at four different MOI values: 0.1, 1, 10 and 100. Depuration in static seawater at multiplicity of infection (MOI) of 0.1 with single phage suspensions of phSE-2 and phSE-5 provided the best results, as it decreased by ~1.3 and 1.7 logCFU/g, respectively, the concentration of Salmonella spp. after a 4h treatment. At a MOI of 0.1, the rate of inactivation with single phage suspensions was higher when compared with the results obtained using the phage cocktail. However, in naturally contaminated cockles treated in static seawater with single phage suspensions and phage cocktail phSE-2/phSE-5, similar decreases in cultivable bacteria concentration (~0.7-0.9 logCFU/g) were achieved after 6h of treatment. When artificially contaminated cockles were depurated with phage phSE-5 in a recirculated seawater system (mimicking industrial depuration conditions), a 0.9 and 2.0 logCFU/g reduction of Salmonella spp. was reached after 4 and 6h treatment. Once the depuration process was performed without phage, a 6h treatment was needed to obtain a 1.1 logCFU/g reduction of Salmonella spp. Results indicated that combining phage biocontrol with depuration procedures enhance bivalve microbial safety for human consumption by improving decontamination efficiency, proving that this technology can be transposed to the bivalves industry. Moreover, this approach also displays the advantage of reducing the time required for depuration and consequently its associated costs.