Methionine and Tryptophan Play Different Modulatory Roles in the European Seabass (Dicentrarchus labrax) Innate Immune Response and Apoptosis Signaling—An In Vitro Study

The range of metabolic pathways that are dependent on a proper supply of specific amino acids (AA) unveils their importance in the support of health. AA play central roles in key pathways vital for immune support and individual AA supplementation has shown to be able to modulate fish immunity. In vitro trials are important tools to evaluate the immunomodulatory role of AA, and the present study was conceived to evaluate methionine and tryptophan roles in immune-related mechanisms aiming to understand their effects in leucocyte functioning and AA pathways. For that purpose, head-kidney leucocytes were isolated and a primary cell culture established. The effect of methionine or tryptophan surplus on cell viability was assessed. Medium L-15 10% FBS without AA addition (0.5mM of L-methionine, 0.1 mM of L-tryptophan) was used as control. To that, L-methionine or L-tryptophan were supplemented at 1 and 2 times (M1x or M2x, and T1x or T2x). Nitric oxide, ATP, total antioxidant capacity, and immune-related genes were evaluated in response to lipopolysaccharides extracted from Photobacterium damselae subsp. piscicida or UV-inactivated bacteria). Moreover, caspase 3 activity and apoptosis-related genes were evaluated in response to the apoptosis-inducing protein, AIP56. Distinct roles in leucocytes’ immune response were observed, with contrasting outcomes in the modulation of individual pathways. Methionine surplus improved cell viability, polyamine production, and methionine-related genes expression in response to an inflammatory agent. Also, methionine supplementation lowered signals of apoptosis by AIP56, presenting lower caspase 3 activity and higher il1ß and nf-¿b expression. Cells cultured in tryptophan supplemented medium presented signals of an attenuated inflammatory response, with decreased ATP and enhanced expression of anti-inflammatory and catabolism-related genes in macrophages. In response to AIP56, leucocytes cultured in a tryptophan-rich medium presented lower resilience to the toxin, higher caspase 3 activity and expression of caspase 8, and lower expression of several genes, including nf-¿b and p65. This study showed the ability of methionine surplus to improve leucocytes’ response to an inflammatory agent and to lower signals of apoptosis by AIP56 induction, while tryptophan attenuated several cellular signals of the inflammatory response to UV-inactivated bacteria and lowered leucocyte resilience to AIP56.This work was partially supported by UIDB/04423/2020, UIDP/ 04423/2020 and INFLAMMAA (reference PTDC/CVT-CVT/ 32349/2017), financed by Portugal and the European Union through FEDER and COMPETE 2020, and national funds through Fundação para a Ciência e a Tecnologia (FCT, Portugal). MM and BC were supported by FCT, Portugal (SFRH/BD/108243/2015 and IF/00197/2015, respectively)

Bayesian spatio-temporal CPUE standardization: Case study of European sardine (Sardina pilchardus) along the western coast of Portugal

Understanding the key factors influencing population dynamics of fish stocks requires knowledge of their spatial distribution and seasonal habitat selection, but these spatio-temporal dynamics are often not explicitly included in ecological studies and stock assessment models. This study standardized the data of sardine fishery-dependent catch-per- unit- effort (CPUE) from the west coast of Portugal using Bayesian hierarchical spatio-temporal models (BHSTM) with the integrated nested Laplace approximation (INLA). Sardine CPUE was best explained by length of the vessel, vessel ID, month, year, and location (latitude, longitude). In terms of spatio-temporal distribution, sardine biomass prediction maps showed a constant pattern that changed every quarter of the year. In addition, sardine CPUE index showed a cyclical trend along the year with minimum values in July and maximum peak in November. This approach provided insights on variables and corresponding modelling effects that may be relevant in spatio-temporal fishery-dependent data standardization, and that could be applied to other fish species and areas.En prens

Bayesian spatio-temporal CPUE standardization: case study of European sardine (Sardina pilchardus) along the western coast of Portugal

Fishery data is one of the most accessible sources of information currently used for ecological studies and stock assessments. Unlike scientific surveys that are usually restricted to a given time of the year, fisheries dependent data is almost continuously available in time. Moreover, the information collected from the fisheries is less expensive and time consuming. However, for use as a relative abundance index, fishery-dependent data requires standardization as catch-per-unit-effort (CPUE) in order to remove the impact of vessel-specific differences and fishing behavior. Understanding the key factors that influence the population dynamics of fish species implies assessment of their spatial distribution and seasonal habitat selection but, spatio-temporal dependence issues are often not explicitly included in the modeling process. This study standardizes sardine fishery-dependent data obtained from the west coast of Portugal as CPUE by means of a Bayesian hierarchical spatio-temporal model using integrated nested Laplace approximation (INLA). This is one of the first studies of the region to provide maps of the relative abundance of this species for all months of the year. The best model included length of the vessel, vessel ID, month, year and location (latitude, longitude), while none of the five environmental covariates (Chl-a, SST, bathymetry, current velocity and direction) were relevant. In terms of spatial distribution, sardines were more abundant in the northern area, especially during the last quarter of the year. The applied methodology has contributed to improve our knowledge of European sardine distribution throughout the year, providing accurate predictive maps and insights into the standardization process of fishery-dependent data that could also be applied to other fish species and areas

Development of New Canned Chub Mackerel Products Incorporating Edible Seaweeds—Influence on the Minerals and Trace Elements Composition

This study aimed to develop new canned chub mackerel products incorporating edible seaweeds (Ascophyllum nodosum, Fucus spiralis, Saccorhiza polyschides, Chondrus crispus, Porphyra sp. and Ulva sp.) harvested in the Portuguese North-Central coast, with simultaneous sensory improvement and minerals enrichment. Two processes were compared, namely the addition of seaweeds in i) the canning step and ii) in the brining step (as the replacement for salt). The concentrations of four macrominerals (Na, K, Ca and Mg), chloride, and twelve trace elements (Co, Cu, Fe, I, Li, Mn, Mo, Rb, Se, Sr, V and Zn) were determined by high-resolution continuum source flame atomic absorption spectrometry (HR-CS-FAAS) and inductively coupled plasma mass spectrometry (ICP-MS), respectively. Results showed that canned chub mackerel incorporating C. crispus and F. spiralis was found to be the preferred sensory option, also exhibiting contents enriched with Cl, Co, Cu, Fe, I, Li, Mg, Mn, Mo, Na, Rb, Se, and Sr. This effect was more pronounced when both seaweed species were added to replace the salt added in the brining step.This work received financial support from project iCanSea—Conservas com macroalgas para diferenciação nutricional e sensorial, sponsored by Portugal 2020 under the Programa Operacional Competitividade e Internacionalização (Ref.3171/Portugal 2020). The Fundação para a Ciência e a Tecnologia (FCT) and the FEDER, under Programme PT2020 (Project UID/QUI/50006/2019) and the project Qualidade e Segurança Alimentar—uma abordagem (nano)tecnológica (NORTE-01-0145-FEDER-000011) are also acknowledged for the financial funding.info:eu-repo/semantics/publishedVersio

Oral vaccination of fish against vibriosis using spore-display technology

Publisher Copyright: Copyright © 2022 Gonçalves, Santos, Coutinho, Pedrosa, Curado, Machado, Costas, Bonneville, Serrano, Carvalho, Díaz-Rosales, Oliva-Teles, Couto and Serra.Oral vaccines are highly demanded by the aquaculture sector, to allow mass delivery of antigens without using the expensive and labor-intensive injectable vaccines. These later require individual handling of fish, provoking stress-related mortalities. One possible strategy to create injection-free vaccine delivery vehicles is the use of bacterial spores, extremely resistant structures with wide biotechnological applications, including as probiotics, display systems, or adjuvants. Bacterial spores, in particular those of Bacillus subtilis, have been shown to behave as mucosal vaccine adjuvants in mice models. However, such technology has not been extensively explored against fish bacterial disease. In this study, we used a laboratory strain of B. subtilis, for which a variety of genetic manipulation tools are available, to display at its spores surface either a Vibrio antigenic protein, OmpK, or the green fluorescence protein, GFP. When previously vaccinated by immersion with the OmpK- carrying spores, zebrafish survival upon a bacterial challenge with V. anguillarum and V. parahaemolyticus, increased up to 50 - 90% depending on the pathogen targeted. Further, we were able to detect anti-GFP-antibodies in the serum of European seabass juveniles fed diets containing the GFP-carrying spores and anti-V. anguillarum antibodies in the serum of European seabass juveniles fed the OmpK-carrying spores containing diet. More important, seabass survival was increased from 60 to 86% when previously orally vaccinated with in-feed OmpK- carrying spores. Our results indicate that B. subtilis spores can effectively be used as antigen-carriers for oral vaccine delivery in fish.publishersversionpublishe

European Sea Bass (Dicentrarchus labrax) immune status and disease resistance are impaired by arginine dietary supplementation

Infectious diseases and fish feeds management are probably the major expenses in the aquaculture business. Hence, it is a priority to define sustainable strategies which simultaneously avoid therapeutic procedures and reinforce fish immunity. Currently, one preferred approach is the use of immunostimulants which can be supplemented to the fish diets. Arginine is a versatile amino acid with important mechanisms closely related to the immune response. Aiming at finding out how arginine affects the innate immune status or improve disease resistance of European seabass (Dicentrarchus labrax) against vibriosis, fish were fed two arginine-supplemented diets (1% and 2% arginine supplementation). A third diet meeting arginine requirement level for seabass served as control diet. Following 15 or 29 days of feeding, fish were sampled for blood, spleen and gut to assess cell-mediated immune parameters and immune-related gene expression. At the same time, fish from each dietary group were challenged against Vibrio anguillarum and survival was monitored. Cell-mediated immune parameters such as the extracellular superoxide and nitric oxide decreased in fish fed arginine-supplemented diets. Interleukins and immune-cell marker transcripts were down-regulated by the highest supplementation level. Disease resistance data were in accordance with a generally depressed immune status, with increased susceptibility to vibriosis in fish fed arginine supplemented diets. Altogether, these results suggest a general inhibitory effect of arginine on the immune defences and disease resistance of European seabass. Still, further research will certainly clarify arginine immunomodulation pathways thereby allowing the validation of its potential as a prophylactic strategy.European Union's Seventh Framework Programme AQUAEXCEL (Aquaculture Infrastructures for Excellence in European Fish Research) [262336]; AQUAIMPROV [NORTE-07-0124-FEDER-000038]; North Portugal Regional Operational Programme (ON. 2 - O Novo Norte) , under the National Strategic Reference Framework, through the European Regional Development Fund; North Portugal Regional Operational Programme (ON. 2 - O Novo Norte), under the National Strategic Reference Framework through the COMPETE - Operational Competitiveness Programme; Fundacao para a Ciencia e Tecnologia; Fundacao para a Ciencia e Tecnologia [SFRH/BD/89457/2012, SFRH/BPD/77210/2011]; Generalitat Valenciana through the project REVIDPAQUA [ISIC/2012/003]; [PEst-C/MAR/LA0015/2013]; [UID/Multi/04423/2013]info:eu-repo/semantics/publishedVersio

Evaluation of SMOS L4 Sea Surface Salinity Product in the Western Iberian Coast

Special issue Moving Forward on Remote Sensing of Sea Surface Salinity.-- 24 pages, 14 figures, supplementary materials https://www.mdpi.com/article/10.3390/rs14020423/s1.-- Data Availability Statement: Data sharing not applicableSalinity is one of the oldest parameters being measured in oceanography and one of the most important to study in the context of climate change. However, its quantification by satellite remote sensing has been a relatively recent achievement. Currently, after over ten years of data gathering, there are still many challenges in quantifying salinity from space, especially when it is intended for coastal environments study. That is mainly due to the spatial resolution of the available products. Recently, a new higher resolution (5 km) L4 SMOS sea surface salinity (SSS) product was developed by the Barcelona Expert Center (BEC). In this study, the quality of this product was tested along the Western Iberian Coast through its comparison with in situ observations and modelled salinity estimates (CMEMS IBI Ocean Reanalysis system). Moreover, several parameters such as the temperature and depth of in situ measurements were tested to identify the variables or processes that induced higher errors in the product or influenced its performance. Lastly, a seasonal and interannual analysis was conducted considering data between 2011 to 2019 to test the product as a potential tool for long-term studies. The results obtained in the present analysis showed a high potential of using the L4 BEC SSS SMOS product in extended temporal and spatial analyses along the Portuguese coast. A good correlation between the satellite and the in situ datasets was observed, and the satellite dataset showed lower errors in retrieving coastal salinities than the oceanic model. Overall, the distance to the coast and the closest rivers were the factors that most influenced the quality of the product. The present analysis showed that great progress has been made in deriving coastal salinity over the years and that the SMOS SSS product is a valuable contribution to worldwide climatological studies. In addition, these results reinforce the need to continue developing satellite remote sensing products as a global and cost-effective methodology for long-term studiesThis work was conducted within the framework of the project AQUIMAR—Marine Knowledge Supporting Aquaculture (MAR-02.01.01-FEAMP-0107), funded by the Mar 2020—Operational Program Mar2020. B.B. was funded by a grant from Mar2020 under AQUIMAR project and also by a PhD grant awarded by Fundação para a Ciência e a Tecnologia (FCT) within the scope of the MIT Portugal Program. A.C.B. was funded by FCT through the Scientific Employment Stimulus Programme (CEECIND/0095/2017). A.T. was funded by Project SARDINHA2020 (MAR-01.04.02-FEAMP-0009), funded by the Operational Program Mar2020. This work benefited from the Infrastructure CoastNet (http://geoportal.coastnet.pt, accessed on 30 September 2021), funded by FCT and the European Regional Development Fund (FEDER), through LISBOA2020 and ALENTEJO2020 regional operational programs, in the framework of the National Roadmap of Research Infrastructures of strategic relevance (PINFRA/22128/2016). This study also received further support from FCT through MARE’s strategic program (UID/MAR/04292/2019). This work represents a contribution to CSIC Thematic Interdisciplinary Platform PTI Teledetect, with the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S). This publication was also funded by the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement N810139: Project Portugal Twinning for Innovation and Excellence in Marine Science and Earth Observation—PORTWIMSPeer reviewe

Diabetes as an independent predictor of high atherosclerotic burden assessed by coronary computed tomography angiography: The coronary artery disease equivalent revisited

(1) To study the prevalence and severity of coronary artery disease (CAD) in diabetic patients. (2) To provide a detailed characterization of the coronary atherosclerotic burden, including the localization, degree of stenosis and plaque composition by coronary computed tomography angiography (CCTA). Single center prospective registry including a total of 581 consecutive stable patients (April 2011-March 2012) undergoing CCTA (Dual-source CT) for the evaluation of suspected CAD without previous myocardial infarction or revascularization procedures. Different coronary plaque burden indexes and plaque type and distribution patterns were compared between patients with (n = 85) and without diabetes (n = 496). The prevalence of CAD (any plaque; 74.1 vs. 56 %; p = 0.002) and obstructive CAD (≥50 % stenosis; 31.8 vs. 10.3 %; p<0.001) were significantly higher in diabetic patients. The remaining coronary atherosclerotic burden indexes evaluated (plaque in LM-3v-2v with prox. LAD; SIS; SSS; CT-LeSc) were also significantly higher in diabetic patients. In the per segment analysis, diabetics had a higher percentage of segments with plaque in every vessel (2.6/13.1/7.5/10.5 % for diabetics vs. 1.4/7.1/3.3/4.4 % for nondiabetics for LM, LAD, LCx, RCA respectively; p<0.001 for all) and of both calcified (19.3 vs. 9.2 %, p<0.001) and noncalcified or mixed types (14.4 vs. 7.0 %; p<0.001); the ratio of proximal-to-distal relative plaque distribution (calculated as LM/proximal vs. mid/distal/branches) was lower for diabetics (0.75 vs. 1.04; p = 0.009). Diabetes was an independent predictor of CAD and was also associated with more advanced CAD, evaluated by indexes of coronary atherosclerotic burden. Diabetics had a significantly higher prevalence of plaques in every anatomical subset and for the different plaque composition. In this report, the relative geographic distribution of the plaques within each subgroup, favored a more mid-to-distal localization in the diabetic patients