Unlocking the in vitro anti-Trypanosoma cruzi activity of halophyte plants from the southern Portugal

Objective:To evaluate the in vitro anti-Trypanosoma cruzi（T.cruzi） activity of organic extracts prepared from halophyte species collected in the southern coast of Portugal（Algarve）,and chemically characterize the most active samples.Methods:Acetone,dichloromethane and methanol extracts were prepared from 31 halophyte species and tested in vitro against trypomastigotes and intracellular amastigotes of the Tulahuen strain of T.cruzi.The most active extract was fractionated by preparative HPLC-DAD,affording 11 fractions.The most selective fraction was fully characterized by 1H-NMR.Results:From 94 samples tested,one was active,namely the root dichloromethane extract of Juncus acutus(IC50 < 20 μg/mL).This extract was fractionated by HPLC,affording 11 fractions,one of them containing only a pure compound（juncunol）,and tested for anti-parasitic activity.Fraction 8(IC50 = 4.1 μg/mL) was the most active,and was further characterized by 1H-NMR.The major compounds were phenanthrenes,9,10-dihydrophenanthrenes and benzocoumarins.Conclusion:Our results suggest that the compounds identified in fraction 8 are likely responsible for the observed anti parasitic activity.Further research is in progress aiming to isolate and identify the specific active molecules.To the best of our knowledge,this is the first report on the in vitro anti T.cruzi activity of halophyte species.

Re-evaluation of Rostraria bierii Tokioka,1970(Annelida)from Seto,Japan as a magelonid,with a review of the Magelonidae of the western Pacific

The identity of Rostraria bierii,originally described as a larval amphinomid from Cape Setozaki,Pacific coast of Japan,is investigated.Based on the original description and illustrations,reinterpretations conclude the“larva”to represent a partial juvenile or adult magelonid specimen,broken after the first chaetiger.The original figures are compared with several known magelonid species to justify the new placement.The authors suggest the supposed amphinomid larva is a Magelonidae taxon inquirendum.The identity of the species is discussed in line with the current knowledge of the Magelonidae in the western Pacific and a key to all known species within the region is provided to aid identifications.Current gaps in our taxonomic knowledge of the Magelonidae of the western Pacific are highlighted and discussed.

Mesocosm Hatcheries Using Semi-intensive Methodologies and Species Diversification in Aquaculture

Mesocosm hatcheries using semi-intensive culture methodologies are characterized by large tanks （40 m3） for fish larvae production at low densities （2-8 larvae/L）, with live prey production within the tank supplemented by exogenous sources of feed. Due to the high biological quality of larvae produced and low level of technological input, these methodologies are regarded as an interesting option to meet the increasing fry demand from the aquaculture industry. This study aimed to describe the experience at Centro de Maricultura da Calheta, Madeira Island, Portugal, with the establishment of a mesocosm hatchery for the production of marine fish larvae. Production trials were initiated with gilthead seabream Sparus aurata. This larval species presented faster growth in total body length （TL） per day （TL = 3.7516e~ ~293day, R2 = 0.9404）, higher survival rate （SR = 31.9%） and comparable swimbladder inflation rates （SB = 90%） to those of larvae reared with intensive methodologies. High larval performance was obtained with two candidate species for aquaculture： the red porgy Pagruspagrus （TL = 3.212e~~39day, R2 = 0.995; SR = 15.3%; SB = 95.7%） and the white seabream Diplodus sargus （TL = 3.6355e~~413day, R2= 0.9824; SR = 25%; SB --- 100%）. Testing ofa 110 m3 mesocosm rearing tank with white seabream larvae revealed limitations associated to larger tanks namely, the amount and quality of live feed required daily. First trials with striped jack （Pseudocararoc dentex） larvae resulted in low larval survival （0.04%）, but the use of a white tank avoided culture management constraints related to positive phototropism. The present results suggest that a mesocosm using semi-intensive methodologies may contribute to species diversification and consequently, to the sustainable development of aquaculture.

somatostatin1基因突变斑马鱼仔鱼转录组分析

利用CRISPR/Cas9基因编辑技术在斑马鱼中构建了可稳定遗传的生长抑素(somatostatin,SST)基因(sst1)突变体鱼系。通过比较6 dpf(days post fertilization,dpf)的sst1^(-/-)突变体和野生型仔鱼转录组,发现sst1^(-/-)突变体相关生化过程和信号通路发生了显著变化。相比野生型,sst1^(-/-)突变体仔鱼中有354个基因显著上调,504个基因显著下调。GO(gene ontology)富集分析表明,大部分差异基因与氨基酸和蛋白质生物合成相关。KEGG(kyoto encyclopedia of genes and genomes)通路富集显示代谢通路、氨基酸合成、氨基酰基-tRNA合成、核糖体合成和rRNA加工相关通路表达增强。结果表明,sst1^(-/-)突变体蛋白质生物合成和代谢过程更加活跃。RT-qPCR验证结果显示,随机选取的10个基因mRNA表达变化趋势与转录组测序结果一致,说明转录组测序结果真实可靠。6月龄突变体体质量、体长和野生型相比并无显著差异。本研究结果为进一步挖掘生长抑素基因家族的潜在功能奠定基础。

Integrating geochemical(surface waters, stream sediments)and biological(diatoms) approaches to assess AMD environmental impact in a pyritic mining area： Aljustrel(Alentejo, Portugal)

Aljustrel mines were classified as having high environmental hazard due to their large tailings volume and high metal concentrations in waters and sediments.To assess acid mine drainage impacted systems whose environmental conditions change quickly,the use of biological indicators with short generation time such as diatoms is advantageous.This study combined geochemical and diatom data,whose results were highlighted in 3 groups：Group 1,with low p H（1.9–5.1）and high metal/metalloid（Al,As,Cd,Co,Cu,Fe,Mn,Ni,Pb,Zn;0.65–1032 mg/L）and SO4（405–39124 mg/L）concentrations.An acidophilic species,Pinnularia aljustrelica,was perfectly adapted to the adverse conditions;in contrast,teratological forms of Eunotia exigua were found,showing that metal toxicity affected this species.The low availability of metals/metalloids in sediments of this group indicates that metals/metalloids of the exchangeable fractions had been solubilized,which in fact enables metal/metalloid diatom uptake and consequently the occurrence of teratologies;Group 2,with sites of near neutral p H（5.0–6.8）and intermediate metal/metalloid（0.002–6 mg/L）and SO4（302–2179 mg/L）concentrations;this enabled the existence of typical species of uncontaminated streams（Brachysira neglectissima,Achnanthidium minutissimum）;Group 3,with samples from unimpacted sites,showing low metal/metalloid（0–0.8 mg/L）and SO4（10–315 mg/L）concentrations,high pH（7.0–8.4）and Cl contents（10–2119 mg/L）and the presence of brackish to marine species（Entomoneis paludosa）.For similar conditions of acidity,differences in diversity,abundance and teratologies of diatoms can be explained by the levels of metals/metalloids.

Automatic analysis of deep-water remotely operated vehicle footage for estimation of Norway lobster abundance

Underwater imaging is being used increasingly by marine biologists as a means to assess the abundance of marine resources and their biodiversity. Previously, we developed the first automatic approach for estimating the abundance of Norway lobsters and counting their burrows in video sequences captured using a monochrome camera mounted on trawling gear. In this paper, an alternative framework is proposed and tested using deep-water video sequences acquired via a remotely operated vehicle. The proposed framework consists of four modules:(1) preprocessing,(2) object detection and classification,(3) object-tracking, and(4) quantification. Encouraging results were obtained from available test videos for the automatic video-based abundance estimation in comparison with manual counts by human experts(ground truth). For the available test set, the proposed system achieved 100%precision and recall for lobster counting, and around 83% precision and recall for burrow detection.

Somatostatin 4 regulates growth and modulates gametogenesis in zebrafish

Somatostatin(SST)plays important roles in growth and development.In teleost fishes six SST encoding genes(sst1 to sst6)have been identified although few studies have addressed their function.Here we aim to determine the function of the teleost specific sst4 in the zebrafish.A CRISPR/Cas9 sst4 zebrafish mutant with loss of function(sst4−/−)was produced which grew significantly faster and was heavier at the onset of gonadal maturation than the wild type(WT).Consistent with their faster growth,liver igf1,igf2a and igf2b expression was significantly upregulated in the sst4−/−fish compared to the WT.Histological examination of the ovaries and testis indicated that sst4−/−fish had slightly delayed testicular gametogenesis compared to the WT.Significantly lower expression of igf3,amh,insl3,hsd17b3,hsd11b2,hsd20b,cyp11b and cyp17 was consistently observed in the sst4−/−testis.In contrast,the ovaries had lower expression of igf1,igf2a and cyp19a1a but increased expression of igf2b and hsd20b.The gonadotrophin beta subunits(fshb and lhb)in the brain were downregulated indicating the brain-pituitary-gonadal axis was downregulated in the sst4−/−fish and suggesting that the steroid production is compromised in the maturing gonads.In addition,analysis of sst1 and sst3 mRNA levels in sst4−/−fish suggests a dosage compensation effect of sst1 in the brain and liver.Altogether,the results from the zebrafish sst4−/−line support the idea that sst4 is involved in the regulation of igf signalling,somatic growth and reproduction since steroidogenesis and gametogenesis at pubertal onset were compromised.

Uncovering the immunological repertoire of the carpet shell clam Ruditapes decussatus through a transcriptomic-based approach

The grooved carpet-shell clam Ruditapes decussatus is native to the Northern Atlantic and Mediterranean Sea and has a high commercial value.It is one of the main native bivalve species cultured in Europe.The main objective of the present study was to gain further insights into the immunological repertoire of R.decussatus through a transcriptomic approach.Pooled mantle samples of eight R.decussatus individuals were sequenced using Illumina platform.A total of 67132 contigs with more than 800 bp were obtained.Manual annotation of these contigs revealed 146 immune-related genes.The gene families in which the highest number of immune-related genes was observed were:C1q domain-containing proteins(63),tumor necrosis factors(15)and toll-like receptors(TLRs,10).A total of 5359 putative single nucleotide polymorphisms(SNPs)were identified in the 146 immune-related genes.The density of SNPs ranged between 0.04 and 7.92 SNPs/100 bp.The highest and the lowest SNP density were observed in genes of the C1q domain-containing protein family.Due to the importance of TLRs in innate immunity,we focused our attention on these membrane receptors.Ten TLRs were identified based on protein domain organization.Phylogenetic analysis revealed that R.decussatus TLRs were diverse and only 3 showed orthology with TLRs of known immune functions in other bivalve species.Moreover,our analysis suggests that lineage restricted-expansions of TLRs occurred in all mollusc taxa analysed including in venerids.

Editorial: Marine protected areas and small-scale fisheries

Marine Protected Areas(MPAs)have long been recognized to be effective tools for spatial management,conservation of biodiversity and fisheries enhancement(Rodríguez-Rodríguez,2016;Claudet et al.,2020),leading Member States Parties to the Convention on Biological Diversity(CBD)in 2010 to agree to cover 10%of their coastal and marine areas with MPAs by 2020(Aichi Target 11).In 2020 MPAs cover 5.3%world’s oceans,with 2.5%in no-take marine reserves offering protection,even though a global no-take coverage of at least 30%of the world’s oceans by 2030 has been proposed(Marine Conservation Institute;http://www.mpatlas.org/progress/proposals/).

Fisher's perceptions about a marine protected area over time

The perceptions of fishers towards the Arrábida Marine Park,a marine protected area(MPA)in the west coast of Portugal,were studied through face-to-face interviews in two different moments of the MPA life cycle.Fishers'perceptions about the MPA and the impact it had on the fishing activity over time were identified just before the full implementation of the zoning and regulations of the management plan and 10 years later.This study aimed to investigate fishers'knowledge,acceptance and perceptions about the MPA changed with time,if support for the MPA was linked to the impact of the MPA on the fishing activity,and if fishers'perceptions about the impact of the MPA on the fishing activity match with local landings trends.Results show that although knowledge about the marine park significantly improved over time,fishers'acceptance did not.A decrease on fishers'support was not substantial but occurred.Issues such as the disagreement with regulations reinforced concerns raised during the implementation of the marine park,particularly in relation to the top-down decision-making,which commonly confers minor participation,recognition and legitimacy to fishers.Apparently,fisheries benefits were still not perceived by local fishers,though they are central for fishers'support.Further,the perceived negative impacts of the park seemed to be more related to social aspects and individual interests than to impacts on catches.Addressing adequate management,enforcement and participation of local fishers is still possible and are advocated here as to contribute to the expected socioecological outcomes and respective support,leading to the future successful performance of the Arrábida Marine Park.Assessing fishers’perceptions towards an MPA over time is central and should be included on periodical socioecological monitoring and inform an effective adaptive management.

Assessing potential protection effects on commercial fish species in a Cuban MPA

Cuba has been leading marine protected area(MPA)designation in the Caribbean region to ensure conservation of its valuable marine ecosystems.Yet,an efficient monitoring program for MPAs is still to be implemented and will benefit from pre-existing information.The highly regulated MPA of Punta Francés National Park is one of the first Cuban MPAs and was established mainly to protect species and ecosystems for tourism purposes.Monitoring of protection effects on commercial fish species was lacking in this MPA.This study aimed at increasing local scientific knowledge by providing a baseline study about the most commercially fished families(Haemulidae,Lutjanidae and Serranidae)around Punta Francés MPA.Data collected represent only a limited period but can be used as a base point to support future monitoring.Fish abundance in number and biomass,and size were collected to test for differences between inside and adjacent areas outside the Punta Francés MPA,in different coral reef types.The main result of this study was the significantly larger size and biomass of snappers and groupers outside the MPA where intense fisheries occur.An observation consistent with a large spawning aggregation was also recorded outside the MPA.Even with a limited set of data,these results suggest that,at least temporarily,the most targeted species[and sizes]are still highly vulnerable to fisheries.Relevant habitats in the adjacent area,that are apparently missing within the MPA,may support some of the results found.Extending the limits of the Punta Francés MPA to include those habitats outside may be critical to ensure the effectiveness of this MPA in contributing to protect the most commercial species of the region.This should be done together with efficient fisheries management measures in the region,such as the significantly increase of minimum legal sizes and temporal closures during spawning migrations.

Roots and rhizomes of wild Asparagus:Nutritional composition,bioactivity and nanoencapsulation of the most potent extract

The nutritional composition and bioactive properties of roots and rhizomes of Asparagus stipularis were evaluated.Antioxidant activity of extracts obtained by infusion was evaluated using free radicals scavenging and reducing power methods.Porcine liver primary cell was used to check the hepatotoxicity of infusions.Results revealed that Asparagus samples are likely a source of nutrients,such as dietary fibre and essential fatty acids.HPLC-DAD-ESI/MS characterization of infusions allowed the identification and quantitation of 7 phenolic compounds,all hydroxycinnamoyl derivatives,with caffeic acid as the most abundant.Roots infusion contained the highest amounts of these compounds.It also exhibited the highest antioxidant activity in all assays,with EC_(50) values of 0.44±0.01,0.98±0.03 and 0.64±0.01 mg/mL for DPPH,ABTS and FRAP assays,respectively,with no toxicity towards PLP2 primary cell cultures(GI_(50)>400μg/mL).PLGA nanoparticles loaded with root extract were prepared using solvent-evaporation double emulsion method.Nanoparticles size was about 260 nm and a polydispersity index around 0.1,with a zeta potential of about-36 mV,as well as a good encapsulation efficiency of approximately 83%.Their morphology was analysed by SEM and spherical polymeric nanoparticles with a smooth surface were observed.FTIR and DSC were also performed,which allowed corroborating the efficacy of the encapsulation and to confirm the production of a stable and robust system to load Asparagus extracts.The developed nanoparticles are expected to be used as delivery systems for bioactive compounds of A.stipularis and they could be used as an innovative dietary supplement.