European Journal of Phycology最新文献

{"title":"Physiological differences among cryptic species of the Mediterranean crustose coralline alga Lithophyllum stictiforme (Corallinales, Rhodophyta)","authors":"Sophie Martin, Virgile Calvert, Anne Chenuil","doi":"10.1080/09670262.2023.2281484","DOIUrl":"https://doi.org/10.1080/09670262.2023.2281484","url":null,"abstract":"The crustose coralline alga Lithophyllum stictiforme is an important reef builder, forming coralligenous concretions in the Mediterranean Sea. This algal species complex is composed of eight crypti...","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2023-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139029192","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Factors contributing to the potential expansion of Limnomonas gaiensis (Chlamydomonadales, Chlorophyta) in freshwater lakes in Northern Europe","authors":"Sirje Sildever, Rebecca I.A. Stewart, Sylvie V.M. Tesson","doi":"10.1080/09670262.2023.2274080","DOIUrl":"https://doi.org/10.1080/09670262.2023.2274080","url":null,"abstract":"Limnomonas gaiensis is a recently described green microalga inhabiting freshwater lakes in northern Europe (England, Sweden). There is little information on the species’ tolerance to environmental ...","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2023-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138547965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Non-indigenous seaweeds in the Northeast Atlantic Ocean, the Mediterranean Sea and Macaronesia: a critical synthesis of diversity, spatial and temporal patterns","authors":"Luna M. van der Loos, Quinten Bafort, Samuel Bosch, Enric Ballesteros, Ignacio Bárbara, Estibaliz Berecibar, Aurélie Blanfuné, Kenny Bogaert, Silke Bouckenooghe, Charles-François Boudouresque, Juliet Brodie, Ester Cecere, Pilar Díaz-Tapia, Aschwin H. Engelen, Karl Gunnarson, Soha Hamdy Shabaka, Razy Hoffman, Vivian Husa, Álvaro Israel, Mart Karremans, Jessica Knoop, Line Le Gall, Christine A. Maggs, Frédéric Mineur, Manuela Parente, Frank Perk, Antonella Petrocelli, Conxi Rodríguez-Prieto, Sandrine Ruitton, Marta Sansón, Ester A. Serrão, Adriano Sfriso, Kjersti Sjøtun, Valérie Stiger-Pouvreau, Gwladys Surget, Ergün Taşkin, Thierry Thibaut, Konstantinos Tsiamis, Lotte Van De Weghe, Marc Verlaque, Frédérique Viard, Sofie Vranken, Frederik Leliaert, Olivier De Clerck","doi":"10.1080/09670262.2023.2256828","DOIUrl":"https://doi.org/10.1080/09670262.2023.2256828","url":null,"abstract":"Effective monitoring of non-indigenous seaweeds and combatting their effects relies on a solid confirmation of the non-indigenous status of the respective species. We critically analysed the status...","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2023-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138495940","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biodiversity of Tehran freshwater cyanobacteria and remote sensing analysis of reservoirs","authors":"Yasaman Tavakoli, Fatemeh Mohammadipanah, Saeid Hamzeh, Amir Sedighi","doi":"10.1080/09670262.2023.2261113","DOIUrl":"https://doi.org/10.1080/09670262.2023.2261113","url":null,"abstract":"ABSTRACTAnalysis of cyanobacterial diversity is an important step towards identifying new biological resources for production of bioactive compounds. In this study cyanobacterial surface coverage was assessed through satellite image analysis of Tehran drinking water reservoirs in late summer and early autumn of 2018. We also analysed freshwater cyanobacteria including seven local strains purified from surface water samples. Following a polyphasic taxonomic approach combining morphological and molecular methods the culturable strains were classified into Synechococcales and Oscillatoriales, representing at least three families and four genera. Similarity-based analysis of their partial 16S rRNA genes showed 93.4–97.0% sequence identity among the strains from different ecosystems. Considering 95–96% as the overall cut-off for discriminating bacterial species, strains UTMC6011 and UTMC6013 are likely to be new species. Physiological characterization of the new strains and nine others isolated previously by our group from another freshwater ecosystem led to identification of seven nitrogen-fixing, 11 sulphide-tolerant, three salt-tolerant and three heterotrophic strains. Further taxonomic analyses are needed to support investigation of their biotechnological potential.Highlights The biodiversity and physiology of cyanobacteria in the main drinking water reservoirs of the megacity, Tehran, were studied by remote sensing and sample analysis.Seven cyanobacterial strains were isolated from the two freshwater lakes. Similarity-based analysis of obtained 16S rRNA gene sequence showed 93.4–97.0% sequence similarity with the strains from other ecosystems.Cyanobacterial isolates of these freshwater reservoirs included seven N2-fixing, 11 sulphide-tolerant, three salt-tolerant and three heterotrophic strains.KEYWORDS: Biodiversityfreshwater cyanobacteriafreshwater reservoir ecosystemphylogenypolyphasic approachremote sensing AcknowledgementsThe authors would like to thank Prof. Alexander V. Pinevich, Dr Svetlana G. Averina and Dr Olga Gavrilova for their valuable comments related to the isolation and morphological studies of cyanobacterial strains.Disclosure statementThe authors state the absence of any competing interests.Supplementary materialThe following supplementary material is accessible via the Supplementary Content tab on the article’s online page at https://doi.org/10.1080/09670262.2023.2261113.Supplementary fig. S1 Phylogenetic tree constructed based on 16S rDNA partial sequences. Obtained sequences aligned and phylogenetic tree constructed using ClustalW and MEGA7 softwares, respectively. 55 cyanobacterial sequences were included and three species of Chloroflexus were used as outgroups. A Maximum likelihood algorithm involving 1000 times boot-strap analyses was performed. Asterisks show strains isolated from Tehran freshwaters. * the isolated strains in this study.Supplementary table S1. Main characteristics of Tehran dams.Supplementary table S2. M","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135242253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phylogenetic analysis of invasive genus <i>Lophocladia</i> (Rhodomelaceae, Rhodophyta) reveals synonymy of <i>L. lallemandii</i> with <i>L. trichoclados</i> and first record of <i>L. kuetzingii</i> in the NE Atlantic","authors":"Raül Golo, Emma Cebrian, Pilar Díaz-Tapia, Petra Lucic, Razzy Hoffman, Alga Vergés","doi":"10.1080/09670262.2023.2260443","DOIUrl":"https://doi.org/10.1080/09670262.2023.2260443","url":null,"abstract":"ABSTRACTSpecies identification in red algae poses significant challenges when relying solely on morphological characteristics. Consequently, the absence of molecular information often conceals misidentifications, cryptic diversity and introduced cryptic species. Within the genus Lophocladia, species have traditionally been delineated based on subtle morphological traits. Lophocladia trichoclados and Lophocladia lallemandii have been extensively documented in warm and temperate coastal regions, with the latter recognized as an invasive species in the Mediterranean. However, the molecular relationship between these species remains unexplored. To address this gap, a comprehensive taxonomic reevaluation of Lophocladia was conducted in the NE Atlantic, Mediterranean Sea and Red Sea. Through combined molecular and morphological analyses of 75 specimens, two distinct taxa of Lophocladia were identified within the study area. Sequences of the rbcL plastid gene unequivocally demonstrated that L. lallemandii and L. trichoclados are conspecific. Consequently, we propose the synonymization of L. lallemandii with L. trichoclados, which has nomenclatural priority. We report L. kuetzingii, a potentially introduced species from Australia, for the first time in the Macaronesian region of the North Atlantic. This finding underscores the importance of expanding red algal DNA datasets, as such efforts significantly enhance our ability to detect and discern introduced species. Additionally, this research highlights the existence of taxonomic uncertainties surrounding introduced species, even among those already classified as invasive.highlights Molecular tools reveal the synonymy of Lophocladia lallemandii with L. trichoclados.L. trichoclados is a widely distributed species in the Atlantic, Mediterranean and Red Sea.L. kuetzingii is detected as a cryptic introduced species in the Macaronesian region.KEYWORDS: Algal bloomsCeramialescryptic introductionsinvasive speciesMacaronesiaMediterraneanrbcLRed Seataxonomy AcknowledgementsWe acknowledge L. Le Gall for her assistance during A. Vergés’ visit to the Herbarium of the Natural History Museum Paris (PC) and thank Patrik Frödén as curator of the Botanical Museum herbarium (LD) who facilitated the pictures of type specimens. We also thank John M. Huisman for providing photographs and resolving questions, as Marc Verlaque and Wilson Freshwater for his help in finding unpublished sequences. Razy Hoffman acknowledges Tal Perevolotsky for collecting specimens of Lophocladia trichoclados from the Interuniversity Institution in Eilat, Red Sea. The School of Plant Sciences and Food Security of Tel Aviv University is also acknowledged for the use of their microscopes and cameras. Some of the Australian collections were possible through funding from the Holsworth Wildlife Research Endowment and their sequence data were generated in the H. Verbruggen laboratory at the University of Melbourne, through financial support of a National","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135270735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The pH tolerance range of the airborne species <i>Tetracystis vinatzeri</i> (Chlorophyceae, Chlamydomonadales)","authors":"Sylvie V.M. Tesson, Sirje Sildever","doi":"10.1080/09670262.2023.2260444","DOIUrl":"https://doi.org/10.1080/09670262.2023.2260444","url":null,"abstract":"ABSTRACTTo decipher the dispersal range of the cosmopolitan green microalga Tetracystis vinatzeri, two airborne-collected strains were barcoded using the 18S ribosomal DNA gene (18S) and the mitochondrial cytochrome c oxidase I subunit (COI), and their survival capacities were investigated across a stepwise acidic to alkaline pH gradient. Genetic fingerprinting confirmed the identity of the strains as representing the same biological species. Investigations of the tolerance of T. vinatzeri to pH levels refined the delimitation of its ecological niche. T. vinatzeri showed optimal growth in freshwater habitats at pH 4.3–9.6 and strains could cope with exposure to slightly more acidic (down to 3.8) and alkaline (up to 10.4) conditions. However, organismal survival was drastically affected by exposure to pH < 3 and > 11. Morphological observations showed that T. vinatzeri could better cope with exposure to acidic pH, exhibiting less cell membrane damage. Genetic analysis showed that the two airborne strains shared the same haplotype, suggesting that they either originated from the same air mass or that the haplotype has a widespread distribution. Further phylogeographic and physiological investigations are required to define the spreading dynamic of T. vinatzeri.Highlights Tetracystis vinatzeri can grow at pH from 3.8–10.4.Strains from different habitats showed sexual compatibility.Low genetic diversity and cosmopolitan distribution suggest widespread dispersal.KEYWORDS: Chlorophyceaedispersalecologymicroalgaesurvival AcknowledgementsFlow cytometry was performed at the FACS Core Facility, Aarhus University, Denmark. The authors are grateful to A. Champeau for proofreading the manuscript.Disclosure statementNo potential conflict of interest was reported by the author(s).Supplementary informationThe following supplementary material is accessible via the Supplementary Content tab on the article’s online page at https://doi.org/10.1080/09670262.2023.2260444Supplementary fig. S1. Pigment profile of Tetracystis vinatzeri for chlorophyll and the phycobilins (phycoerythrin and allophycocyanin).Supplementary fig. S2. Cell signal distribution for chlorophyll pigment and size fraction,distributed by pH category: acid (A, pH 1-4), neutral (B, pH 7-9), and basic (C, pH 10-13).Positive signal for chlorophylls (red) and propidium iodide (blue, dead cells).Author contributionsS. Tesson: original concept, culture experiment, data analysis, drafting and editing manuscript; S. Sildever: molecular data, editing manuscript.Data availability statementGenetic sequences are available in GenBank for both 18S (OQ402668-OQ402669) and COI (OQ412808-OQ412809) markers. S3MWC-21 and S3MWC-29 strains are deposited at the Norwegian Culture Collection of Algae (NIVA-CHL 196 and NIVA-CHL 197).Additional informationFundingThis research and S.T. are funded by the European Union’s Horizon 2020 Research and Innovation Programme under the Marie Skłodowska-Curie grant agreement no. [754513]","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135616727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced reducing sugar production and extraction for <i>Chlorella vulgaris</i> in mixotrophic cultivation using high hydrostatic pressure processing and ultrasound","authors":"Sibel Uzuner, Sebnem Kurhan, Gulsun Akdemir Evrendilek","doi":"10.1080/09670262.2023.2248216","DOIUrl":"https://doi.org/10.1080/09670262.2023.2248216","url":null,"abstract":"ABSTRACTAlthough extraction of polysaccharides to convert reducing sugars (RS) from microalgae by acid or alkali pretreatments and enzymatic hydrolysis has been extensively studied, few reports exploring the use of high hydrostatic pressure processing (HHP) and ultrasonication (US) as emerging technologies for the extraction of sugars from microalgae biomass exist. Thus, the present study was conducted to determine the effects of mixotrophic growth and stress conditions (NaNO3 and CO2 concentration and light intensity) on RS and protein accumulation in the unicellular green alga Chlorella vulgaris in addition to optimization of the effectiveness of the sequential applications of HHP and US with dilute acid as well as simultaneous enzymatic saccharification on the production of RS from microalga cells. High light intensity, high CO2 concentration and limited nitrogen concentration promoted RS production. The maximum protein content (0.0683 mg g‒1) was achieved at 0.3 g l‒1 NaNO3 concentration, 7000 μmol photons m‒2 s‒1 and 6 l min‒1 CO2 concentration. The highest RS content of C. vulgaris after 48 h enzymatic saccharification (583.86 ± 13.23 mg g‒1) was obtained at 1% (w/w) acid concentration and 80% amplitude for 30 min with 79.4% RS yield. Combined US-assisted dilute acid pretreatment and enzymatic hydrolysis were also found to be more effective than HHP assisted dilute acid pretreatment and enzymatic saccharification. Therefore, microalgal biomass can be considered a suitable renewable feedstock used in fermentation.Highlights The cultivation period of Chlorella vulgaris was reduced from 25 days to 14 days using mixotrophic growing conditions.Mixotrophic conditions enhanced reducing sugar productivity.Novel extraction techniques enhanced the extraction of reducing sugar from microalgae.KEYWORDS: High hydrostatic pressure processinglight intensitymicroalgamixotrophic cultivationnitrogen starvationultrasonication AcknowledgementsThe authors would like to thank the Innovative Food Technologies Development Application and Research Center (YENIGIDAM) of Bolu Abant Izzet Baysal University (BAIBU) for HHP analyses, and I. Isci for his help with construction of the lab-scale photobioreactor.Disclosure statementNo potential conflict of interest was reported by the author(s).Author contributionsS. Uzuner: conducted a research and investigation process, specifically performed the experiments, or data/evidence collection, formal analysis (application of statistical or mathematical techniques to analyze or synthesize study data), wrote original draft (preparation and/or creation of the published work, specifically writing the initial draft); G.A. Evrendilek: specifically performed the experiments, or data/evidence collection, wrote review and edited specifically critical review, commentary or revision-including pre- or post-publication stages; S. Kurhan: performed the experiments, or data/evidence collection.Additional informationFundingThis work was suppor","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136295571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genetic and phenotypic intra-species diversity of alga <i>Tisochrysis lutea</i> reveals original genetic structure and domestication potential","authors":"Grégory Carrier, Jérémy Berthelier, Agathe Maupetit, Elodie Nicolau, Martial Marbouty, Nathalie Schreiber, Aurélie Charrier, Clémence Carcopino, Laura Leroi, Bruno Saint-Jean","doi":"10.1080/09670262.2023.2249073","DOIUrl":"https://doi.org/10.1080/09670262.2023.2249073","url":null,"abstract":"Oceanic phytoplankton species are generally composed of many strains, with intra-species diversity consisting of genetic and phenotypic variability. Despite its importance in ecological and biotechnological contexts, this intra-species diversity and variation among strains has been little studied. We investigated the intra-species diversity of the microalga Tisochrysis lutea, a haptophyte of the Isochrysidales order. Inter-strain diversity of T. lutea was studied because of the economic importance of the species as a feed in aquaculture and for antioxidant metabolite production, particularly fucoxanthin and other carotenoids, which have health benefits. We analysed Tara Ocean datasets which revealed that T. lutea was present in the Pacific, Atlantic and Indian Oceans but not in the Arctic or Austral Oceans. We next made phenotypic and genotypic comparisons of 11 strains of T. lutea from worldwide algal collections. All strains were cultivated in the same controlled conditions for one week, and several phenotypic traits were measured, notably antioxidant content. In parallel, the genomes of each strain were sequenced, and genetic variants identified. At the genetic and phenotypic levels, the strains were distinct from each other and our analysis revealed natural trait variations of interest in relation to further exploitation in domestication programmes. A large number of genetic variations were identified among the strains, but no major differences in genome size were observed. Moreover, limited genetic structure was observed among these strains, which could be a consequence of the complex life history of species within the Isochrysidales. Our study provides new knowledge on the intra-species diversity that should be considered in future environmental studies and breeding programmes.","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135243163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tracing the introduction of Dictyota acutiloba (Dictyotales, Phaeophyceae) in the Mediterranean Sea, with a reassessment of its geographic distribution","authors":"Soria Delva, C. F. de la Hoz, Quinten Bafort, S. D’hondt, S. Shabaka, Sarah Hamdy Rashedy, A. Sherwood, T. Guy‐Haim, Á. Israel, O. De Clerck","doi":"10.1080/09670262.2023.2214184","DOIUrl":"https://doi.org/10.1080/09670262.2023.2214184","url":null,"abstract":"","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2023-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47945809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Morphological and taxonomic implications of the formation of colonies in Cyclostephanos and Stephanodiscus (Bacillariophyta)","authors":"Konrad Schultz, Mirko Dressler, Olivier Jacques, Marcus Frank, A. Springer, T. Hübener","doi":"10.1080/09670262.2023.2222297","DOIUrl":"https://doi.org/10.1080/09670262.2023.2222297","url":null,"abstract":"","PeriodicalId":12032,"journal":{"name":"European Journal of Phycology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2023-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42713108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}