Opportunities for Seaweed Aquaculture Development in the Azores

Jornadas "Ciência nos Açores – que futuro? Tema Ciências Naturais e Ambiente", Ponta Delgada, 7-8 de Junho de 2013.As macroalgas marinhas têm sido largamente usadas para diversos fins. Nos Açores, várias espécies têm sido usadas tradicionalmente na alimentação humana (e.g. Fucus spiralis, Porphyra spp., Laurencia spp. e Osmundea spp.) e para extracção de compostos com interesse na indústria dos ficocolóides (Pterocladiella capilacea e Gelidium spp.). As exigências no controlo da qualidade e as práticas actuais de colheita de macroalgas marinhas selvagens na Europa levantam preocupações ambientais sérias que tornam premente a necessidade se implementarem métodos de produção de biomassa controlados, como é o caso da aquacultura de macroalgas marinhas. Apesar da importância da exploração sustentável dos recursos marinhos existentes nos Açores, não existe informação sobre a viabilidade do cultivo de macroalgas marinhas no Arquipélago. O conhecimento sobre os requisitos básicos para o cultivo em grande escala das espécies nativas seleccionadas e os locais mais apropriados para a sua implementação está em falta. O objectivo principal do presente projecto é avaliar o potencial de cultivo de espécies de macroalgas marinhas seleccionadas, bem como identificar os métodos de cultivo mais adequados. Os resultados do programa de doutoramento serão de extrema importância quer em termos científicos quer em termos empresariais. Permitirão a transferência de tecnologia para o tecido empresarial regional e para a implementação de empresas de base tecnológica indo ao encontro das futuras políticas de financiamento europeias no âmbito do Programa Europeu Horizonte 2020.ABSTRACT: Seaweeds have a wide range of applications. In the Azores, several species of seaweeds were traditionally used either as food (e.g. Fucus spiralis, Porphyra spp., Laurencia spp. and Osmundea spp.) or for extraction of chemical products (Pterocladiella capilacea e Gelidium spp.). The product quality control requirements and concerns regarding the environmental sustainability of current wild seaweed biomass harvesting practices in Europe demand for controlled seaweed aquaculture. Despite the interest in exploiting Azorean seaweed resources, there is no information on the feasibility of cultivating seaweed in the Azores. Basic knowledge on large scale cultivation requirements of the selected native species is missing. The present project it’s aimed at evaluating the culture potential of selected Azorean species. The resulting outputs will be extremely important for both academic and economic purposes, bringing together the research and the market. Innovative enterprise will benefit from this project and develop technological breakthroughs into viable products with real commercial potential. This main objective is in according with the principal strategy of the Horizon 2020 that will tackle societal challenges after the end of FP7.Fundação para a Ciência e Tecnologi

Short term effects of irradiance on the growth of Pterocladiella capillacea (Gelidiales, Rhodophyta)

Pterocladiella capillacea has been economically exploited for agar extraction in the Azores for many years. Harvesting dropped to a full stop in the early 1990s due to a population collapse, but restarted in 2013. Since then it has been intensively harvested and overexploitation must be prevented, with both sustainable harvesting and effective cultivation practices. This study represents the first attempt to determine optimal conditions for P. capillacea production in the Azores, and evaluates its vegetative growth in two experiments using von Stosch’s medium designed to test entire thallus and tips portions response to different irradiances (30, 70 and 150 μmol photons m¯² s¯¹). The best relative growth rate (RGR) was recorded at 150 μmol photons m¯² s¯¹ for the entire thalli and tips after two-weeks and three-weeks, respectively, indicating that an acclimation period is necessary to assure the growth of this alga under experimental conditions. Higher RGR was obtained at higher irradiance (3.98 ± 2.10% fm day¯¹), but overall, growth rates were low or negative. Epiphytes were a serious problem towards the end of the entire thallus experiments, where Feldmannia irregularis proliferate at all irradiances. Future cultivation approaches complemented with other relevant environmental factors (e.g. pH, photoperiod, salinity), are recommended.FCT – Fundação para a Ciência e Tecnologia projects UID/BIA/00329/2013, 2015 - 2018 and UID/BIA/00329/2019, CIRN (Centro de Investigação de Recursos Naturais, University of the Azores), and CIIMAR (Interdisciplinary Centre of Marine and Environmental Research, Porto, Portugal). RFP was supported by a doctoral grant M3.1.2/F/024/2011, Fundo Regional para a Ciência e Tecnologia.info:eu-repo/semantics/publishedVersio

Efficient microbial bioconversion of brown macroalgae obtained through profitable high-density sea cultivation using modified microbial strains to produce commodity and specialty chemicals: A developing blue chemical industry in Chile

Plant biomass is considered a promising feedstock for large scale sustainable bio-based green chemistry. However, only the use of agricultural or forestry residues is viable, since they do not compete for land with feed crops and have competitive costs. Moreover, carbohydrate recovery from these sources is always difficult due to their high lignin content. Alternatively, macroalgae are competitive sources of carbohydrate-rich biomass not requiring land or fresh water for its production. Macrocystis pyrifera is one of the fastest-growing macroalgal species with high CO2 fixation efficiency, highly-abundant and accessible carbohydrates. We demonstrated that it can be cultured in temperate seas, yielding 124 ton/Ha/yr, and can be economically profitable at a 10-hectare scale 1,2. Microbial and enzymatic algal biomass bioprocessing has been also undertaken by our group. We demonstrated the technical feasibility of producing ethanol at a pilot industrial scale by fermenting algal carbohydrates with a genetically modified Escherichia coli 3. However, ethanol production, even with high productivities, was not commercially viable. To make algal biomass bioconversion profitable, we performed a large metabolic engineering and synthetic biology project to discover combinations of metabolic pathways, regulation, carbohydrate sources –algal or not– and alternative bioproducts that maximize microbial efficiency and commercial viability. Using a genome-scale reconstruction of Saccharomyces cerevisiae’s metabolism, we demonstrated that redox ratio constraints and the preferential use of NADH or NADPH for alginate metabolism were key for S. cerevisiae conversion of alginate:mannitol carbohydrate sources 4. However, yeast use makes chemical processes technically and economically unfeasible for low value products due to their inability to produce extracellular enzymes for alginate lysis. By means of dynamic metabolic models developed for E. coli, we demonstrated that the main metabolic process bottleneck is microbial carbohydrate metabolization and that algal carbohydrate composition is a key determinant of fermentation efficiency. Using a multi-objective optimization strategy focused on microorganism growth, energy levels and redox ratio conservation, we also showed that ethanol production from algal biomass is incompatible with E. coli’s metabolism, due to low energetic and redox efficiencies obtained from alginate using host microorganism metabolic pathways. We then used high-performance parallel computing to develop a metabolic potentiality map for E. coli in which we explored more than 10.000 possible combinations of metabolic pathways that could be built in our strain to convert brown macroalgae carbohydrates with high efficiency, considering the best combinations of knock-outs and overexpressions to be introduced in E. coli’s central metabolic pathways. With this technique, we identified other valuable chemicals, such as succinic, aspartic, gluconic and levulinic acids, and complex aromatic and aliphatic biomolecules can be efficiently produced from Macrocystis with specifically modified strains for each product. The bulk of our research fostering algal feedstock production and industrial bioconversion in Chile will be presented in this work. 1. Buschmann, A. H. et al. The Status of Kelp Exploitation and Marine Agronomy, with Emphasis on Macrocystis pyrifera, in Chile. Advances in Botanical Research 71, 161–188 (2014). 2. Camus, C., Infante, J. & Buschmann, A. H. Overview of 3 year precommercial seafarming of Macrocystis pyrifera along the Chilean coast. Reviews in Aquaculture 10, 543–559 (2018). 3. Camus, C. et al. Scaling up bioethanol production from the farmed brown macroalga Macrocystis pyrifera in Chile. Biofuels, Bioproducts and Biorefining 10, 673–685 (2016). 4. Contador, C. A. et al. Analyzing redox balance in a synthetic yeast platform to improve utilization of brown macroalgae as feedstock. Metabolic Engineering Communications 2, 76–84 (2015)

Growth responses of Macrocystis pyrifera (Laminariales), Southern Chile, juvenile sporophytes to nutrient limitation

1st Mares Conference on Marine Ecosystems Health and Conservation. Olhão, Portugal 17-21 November 2014.Kelp forests represent some of the most conspicuous coastal habitats and today we recognize only one giant kelp species (Macrocystis pyrifera) distributed globally [1, 2]. M. pyrifera is recognized as a perennial kelp species with a low capacity of energy storage, whereas its high productivity is associated the availability of nitrogen from the water column [3]. The relation between M. pyrifera growth and biomass production results from a plastic response of the sporophytes to temporal and spatial variability in nitrogen availability [4, 5]. However, the low storage capacity of giant kelp [6, 7] is clearly disadvantageous during periods of suboptimal environmental conditions; as those that occur seasonally in California and the inland waters of southern Chile. Due to an increased demand for kelp biomass in Chile for the world alginate industry and abalone farming in Chile [8, 9] there is an increased demand of raw material and interest for developing kelp aquaculture technologies [10]. The present study evaluates the effect of different nitrogen availability on the growth and regeneration of juvenile fronds of M. pyrifera sporophytes from southern Chile and explore its consequences for the development of seeding strategies of kelp farming in southern Chile

State of knowledge regarding the potential of macroalgae cultivation in providing climate-related and other ecosystem services

Macroalgae (or seaweed) aquaculture can potentially provide many ecosystem services, including climate change mitigation, coastal protection, preservation of biodiversity and improvement of water quality. Nevertheless, there are still many constraints and knowledge gaps that need to be overcome, as well as potential negative impacts or scale-dependent effects that need to be considered, before macroalgae cultivation in Europe can be scaled up successfully and sustainably. To investigate these uncertainties, the Expert Working Group (EWG) on Macroalgae was established. Its role was to determine the state of knowledge regarding the potential of macroalgae culture in providing climate-related and other ecosystem services (ES) and to identify specific knowledge gaps that must be addressed before harvesting this potential. The methodological framework combined a multiple expert consultation with Delphi process and a Quick Scoping Review (QSR). To analyse the outcome of both approaches, the EWG classified the findings under the categories Political, Environmental, Social, Technical, Economic and Legal (PESTEL approach) and categorised the ES based on the CICES 5.1 classification. Although representative stakeholders from many different disciplines were contacted, the majority of responses to the Delphi process were from representatives of academia or research. While the results of each method differed in many ways, both methods identified the following top six ecosystem services provided by seaweed cultivation: i) provisioning food, ii) provisioning hydrocolloids and feed, iii) regulating water quality, iv) provisioning habitats, v) provisioning of nurseries and vi) regulating climate. Diverse technological knowledge gaps were identified by both methods at all scales of the macroalgae cultivation process, followed by economic and environmental knowledge gaps depending on the method used. Based on suggestions from the expert respondents in the Delphi process, there is a clear need for an European-wide strategy for reducing risks for seaweed producers, providing clear standards and guidelines for obtaining permits, and providing financial support to improve technological innovation, that will ensure consistent quality. Legal (e.g., safety regulations), economic (e.g., lack of demand for seaweeds in many countries) and technological (e.g., production at large scale) constraints represented almost 70% of the total responses in the Delphi process, whereas environmental and technical constraints were more dominant in the literature. The most commonly identified potential negative impacts of macroalgae cultivation both among the expert responses and the reviewed articles were unknown environmental impacts, e.g. to deep sea, benthic and pelagic ecosystems. The present study provides an assessment of the state of knowledge regarding ES provided by seaweed cultivation and identifies the associated knowledge gaps, constraints and potential negative impacts. One of the main hurdles recognised by the EWG was the understanding of ES themselves by the different stakeholders, as well as the issue of scale. Studies providing clear evidence of ES provided by seaweed cultivation and/or valorisation of these services were lacking in the literature, and some aspects, like cultural impact etc. were missing in the responses to the questionnaires during the Delphi process. The issue of scale and scaling-up was omnipresent both in assessing the ES provided by seaweed cultivation and in identifying knowledge gaps, constraints and potential negative impacts. For example, the ES provided will depend on the scale of cultivation, the main technological knowledge gaps were often related to scale of cultivation. Likewise at a large scale of operations, there could be multiple associated potential side effects, which need to be further investigated. Based on the outcomes of this investigation, we provide an outlook with open questions that need to be answered to support the sustainable scaling-up of seaweed cultivation in Europe

Salmon Aquaculture and Antimicrobial Resistance in the Marine Environment

Antimicrobials used in salmon aquaculture pass into the marine environment. This could have negative impacts on marine environmental biodiversity, and on terrestrial animal and human health as a result of selection for bacteria containing antimicrobial resistance genes. We therefore measured the numbers of culturable bacteria and antimicrobial-resistant bacteria in marine sediments in the Calbuco Archipelago, Chile, over 12-month period at a salmon aquaculture site approximately 20 m from a salmon farm and at a control site 8 km distant without observable aquaculture activities. Three antimicrobials extensively used in Chilean salmon aquaculture (oxytetracycline, oxolinic acid, and florfenicol) were studied. Although none of these antimicrobials was detected in sediments from either site, traces of flumequine, a fluoroquinolone antimicrobial also widely used in Chile, were present in sediments from both sites during this period. There were significant increases in bacterial numbers and antimicrobial-resistant fractions to oxytetracycline, oxolinic acid, and florfenicol in sediments from the aquaculture site compared to those from the control site. Interestingly, there were similar numbers of presumably plasmid-mediated resistance genes for oxytetracycline, oxolinic acid and florfenicol in unselected marine bacteria isolated from both aquaculture and control sites. These preliminary findings in one location may suggest that the current use of large amounts of antimicrobials in Chilean aquaculture has the potential to select for antimicrobial-resistant bacteria in marine sediments

Antimicrobial Resistance in Chile and The One Health Paradigm: Dealing with Threats to Human and Veterinary Health Resulting from Antimicrobial Use in Salmon Aquaculture and the Clinic

The emergence and dissemination of antimicrobial-resistant bacteria (ARB) is currently seen as one of the major threats to human and animal public health. Veterinary use of antimicrobials in both developing and developed countries is many-fold greater than their use in human medicine and is an important determinant in selection of ARB. In light of the recently outlined National Plan Against Antimicrobial Resistance in Chile, our findings on antimicrobial use in salmon aquaculture and their impact on the environment and human health are highly relevant. Ninety-five percent of tetracyclines, phenicols and quinolones imported into Chile between 1998 and 2015 were for veterinary use, mostly in salmon aquaculture. Excessive use of antimicrobials at aquaculture sites was associated with antimicrobial residues in marine sediments 8 km distant and the presence of resistant marine bacteria harboring easily transmissible resistance genes, in mobile genetic elements, to these same antimicrobials. Moreover, quinolone and integron resistance genes in human pathogens isolated from patients in coastal regions adjacent to aquaculture sites were identical to genes isolated from regional marine bacteria, consistent with genetic communication between bacteria in these different environments. Passage of antimicrobials into the marine environment can potentially diminish environmental diversity, contaminate wild fish for human consumption, and facilitate the appearance of harmful algal blooms and resistant zoonotic and human pathogens. Our findings suggest that changes in aquaculture in Chile that prevent fish infections and decrease antimicrobial usage will prove a determining factor in preventing human and animal infections with multiply-resistant ARB in accord with the modern paradigm of One Health

Tension-Compression Loading with Chemical Stimulation Results in Additive Increases to Functional Properties of Anatomic Meniscal Constructs

Objective: This study aimed to improve the functional properties of anatomically-shaped meniscus constructs through simultaneous tension and compression mechanical stimulation in conjunction with chemical stimulation. Methods: Scaffoldless meniscal constructs were subjected to simultaneous tension and compressive stimulation and chemical stimulation. The temporal aspect of mechanical loadingwas studied by employing two separate five day stimulation periods. Chemical stimulation consisted of the application of a catabolic GAG-depleting enzyme, chondroitinase ABC (C-ABC), and an anabolic growth factor, TGF-b1. Mechanical and chemical stimulation combinations were studied through a full-factorial experimental design and assessed for histological, biochemical, and biomechanical properties following 4 wks of culture. Results: Mechanical loading applied from days 10–14 resulted in significant increases in compressive, tensile, and biochemical properties of meniscal constructs. When mechanical and chemical stimuliwere combined significant additive increases in collagen per wet weight (4-fold), compressive instantaneous (3-fold) and relaxation (2-fold) moduli, and tensile moduli in the circumferential (4-fold) and radial (6-fold) directions were obtained. Conclusions: This study demonstrates that a stimulation regimen of simultaneous tension and compression mechanical stimulation, C-ABC, and TGF-b1 is able to create anatomic meniscus constructs replicating the compressive mechanica

2021 Taxonomic update of phylum Negarnaviricota (Riboviria: Orthornavirae), including the large orders Bunyavirales and Mononegavirales.

Correction to: 2021 Taxonomic update of phylum Negarnaviricota (Riboviria: Orthornavirae), including the large orders Bunyavirales and Mononegavirales. Archives of Virology (2021) 166:3567–3579. https://doi.org/10.1007/s00705-021-05266-wIn March 2021, following the annual International Committee on Taxonomy of Viruses (ICTV) ratification vote on newly proposed taxa, the phylum Negarnaviricota was amended and emended. The phylum was expanded by four families (Aliusviridae, Crepuscuviridae, Myriaviridae, and Natareviridae), three subfamilies (Alpharhabdovirinae, Betarhabdovirinae, and Gammarhabdovirinae), 42 genera, and 200 species. Thirty-nine species were renamed and/or moved and seven species were abolished. This article presents the updated taxonomy of Negarnaviricota as now accepted by the ICTV.This work was supported in part through Laulima Government Solutions, LLC prime contract with the US National Institute of Allergy and Infectious Diseases (NIAID) under Contract No. HHSN272201800013C. J.H.K. performed this work as an employee of Tunnell Government Services (TGS), a subcontractor of Laulima Government Solutions, LLC under Contract No. HHSN272201800013C. This work was also supported in part with federal funds from the National Cancer Institute (NCI), National Institutes of Health (NIH), under Contract No. 75N91019D00024, Task Order No. 75N91019F00130 to I.C., who was supported by the Clinical Monitoring Research Program Directorate, Frederick National Lab for Cancer Research. This work was also funded in part by Contract No. HSHQDC-15-C-00064 awarded by DHS S&T for the management and operation of The National Biodefense Analysis and Countermeasures Center, a federally funded research and development center operated by the Battelle National Biodefense Institute (V.W.); and NIH contract HHSN272201000040I/HHSN27200004/D04 and grant R24AI120942 (N.V., R.B.T.). S.S. acknowledges partial support from the Special Research Initiative of Mississippi Agricultural and Forestry Experiment Station (MAFES), Mississippi State University, and the National Institute of Food and Agriculture, US Department of Agriculture, Hatch Project 1021494. Part of this work was supported by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001030), the UK Medical Research Council (FC001030), and the Wellcome Trust (FC001030).S