Márcio Thomaz dos Santos Varjão , Alysson Wagner Fernandes Duarte , Luiz Henrique Rosa , Magna Suzana Alexandre-Moreira , Aline Cavalcanti de Queiroz
{"title":"Leishmanicidal activity of fungal bioproducts: A systematic review","authors":"Márcio Thomaz dos Santos Varjão , Alysson Wagner Fernandes Duarte , Luiz Henrique Rosa , Magna Suzana Alexandre-Moreira , Aline Cavalcanti de Queiroz","doi":"10.1016/j.fbr.2022.01.001","DOIUrl":"10.1016/j.fbr.2022.01.001","url":null,"abstract":"<div><p><span><span>The genome mining<span> of biosynthetic genes from fungi demonstrates the enormous pharmacological potential that is still little explored. These results have encouraged the scientific community to invest in fungi as a source of innovative alternatives for the treatment of neglected diseases, such as leishmaniasis. Therefore, this work aimed to identify, through a systematic search in the databases of PubMed, Lilacs and Scielo, the existing evidence in the literature regarding the efficacy of the leishmanicidal activity of fungal </span></span>bioproducts that represent new starting points for the advancement of pharmacotherapy of leishmaniasis. During the search process, 59 articles met all the eligibility criteria and, therefore, were included in this review. The studies demonstrate that different prospecting, cultivation, biotechnological and synthetic modification strategies contribute to the discovery and development of new therapeutic fungal compounds. 39 (66.1%) of the studies presented at least one isolated compound with leishmanicidal activity, while 20 (33.9%) evaluated only crude extracts or semipurified fractions. Terpenes, steroids and quinones were the most prevalent chemical classes among the isolated compounds. There are many studies about active compounds that have been isolated from </span><span><em>Penicillium</em></span> and <span><em>Aspergillus</em></span> genera. A large majority (89.8%) of the selected studies been conducted <em>in vitro</em>. Only six studies performed <em>in vivo</em> assay. The species of <span><em>Leishmania amazonensis</em></span> and <span><em>Leishmania donovani</em></span> were the most evaluated. The results support the hypothesis of the pharmacological potential of fungal bioproducts in the treatment of leishmaniasis.</p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"40 ","pages":"Pages 91-113"},"PeriodicalIF":6.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43677402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cyril Henard , Marcia R. Saraiva , Magdalena E. Ściślak , Tahmina Ruba , Debbie McLaggan , Patricia Noguera , Pieter van West
{"title":"Can Ulcerative Dermal Necrosis (UDN) in Atlantic salmon be attributed to ultraviolet radiation and secondary Saprolegnia parasitica infections?","authors":"Cyril Henard , Marcia R. Saraiva , Magdalena E. Ściślak , Tahmina Ruba , Debbie McLaggan , Patricia Noguera , Pieter van West","doi":"10.1016/j.fbr.2022.02.002","DOIUrl":"10.1016/j.fbr.2022.02.002","url":null,"abstract":"<div><p>Ulcerative dermal necrosis (UDN), a chronic skin condition, affects primarily mature wild salmonids returning from the sea to freshwater for their spawning. The involvement of water moulds such as <em>Saprolegnia parasitica</em> as a secondary pathogen in this disease is clear but the identification of a primary cause or of primary pathogen(s) remains elusive. In this opinion article, we re-visit UDN regarding epidemiology, pathology and aetiology and speculate the potential involvement of UV radiation in the initiation of UDN in salmonid fish returning from the sea.</p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"40 ","pages":"Pages 70-75"},"PeriodicalIF":6.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1749461322000094/pdfft?md5=f279db2898378d932701a38674be797c&pid=1-s2.0-S1749461322000094-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42891205","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Siqiao Chen , Paul Daly , Dongmei Zhou , Jingjing Li , Xiaoyu Wang , Sheng Deng , Hui Feng , Chunting Wang , Taha Majid Mahmood Sheikh , Yifan Chen , Taiqiang Xue , Feng Cai , Christian P. Kubicek , Lihui Wei , Irina S. Druzhinina
{"title":"The use of mutant and engineered microbial agents for biological control of plant diseases caused by Pythium: Achievements versus challenges","authors":"Siqiao Chen , Paul Daly , Dongmei Zhou , Jingjing Li , Xiaoyu Wang , Sheng Deng , Hui Feng , Chunting Wang , Taha Majid Mahmood Sheikh , Yifan Chen , Taiqiang Xue , Feng Cai , Christian P. Kubicek , Lihui Wei , Irina S. Druzhinina","doi":"10.1016/j.fbr.2022.03.001","DOIUrl":"10.1016/j.fbr.2022.03.001","url":null,"abstract":"<div><p><span><em>Pythium</em></span> species are devasting pathogens causing major crop losses, <em>e.g.,</em><span> damping-off in sugar beet caused by </span><span><em>Pythium ultimum</em></span> and root-rot of tomato caused by <span><em>Pythium aphanidermatum</em></span>. The use of natural antagonistic microorganisms is a promising environment-friendly approach to control <em>Pythium</em>-caused plant diseases. There are several examples of biocontrol of diseases caused by <em>Pythium</em><span> species but the application of bioeffectors (biological control agents) is limited for various reasons, including the restricted amount of gene-modification based biotechnological progress. The regulations in many countries prevent genetically modified bioeffectors from being routinely deployed in field conditions. Our two connected aims in this review are (1) to compile and assess achievements in genetic modification of bioeffectors which have been tested for parasitism or antagonism towards a </span><em>Pythium</em><span> plant pathogen or biocontrol of a plant disease caused by a </span><em>Pythium</em> species, and (2) discuss how a better performing bioeffector could be engineered to improve biocontrol of <em>Pythium</em><span><span><span><span>-caused plant diseases. We focus on the role of seven key mechanisms: cellulases<span>, carbon catabolite de-repression, glycosylation, reactive oxygen species, </span></span>chitin re-modelling, proteases, and toxic </span>secondary metabolites<span>. Genetic modifications of bioeffectors include gene deletion and overexpression, as well as the replacement of promoter elements to tune the gene expression to the presence of the pathogen. Gene-modifications are limited to fungal and bacterial bioeffectors due to the difficulty of gene modification in </span></span>oomycete bioeffectors such as </span><em>Pythium oligandrum</em>. We assess how previous gene modifications could be combined and what other gene modification techniques could be introduced to make improved bioeffectors for <em>Pythium</em>-caused plant diseases. The broad host-range of <em>Pythium</em> spp. suggests engineering improved antagonistic traits of a bioeffector could be more effective than engineering plant-mediated traits <em>i.e.</em>, engineer a bioeffector to antagonise a plant pathogen in common with multiple plant hosts rather than prime each unique plant host.</p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"40 ","pages":"Pages 76-90"},"PeriodicalIF":6.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45319052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Maryam Hashemi , Dania Tabet , Murilo Sandroni , Clara Benavent-Celma , Jenifer Seematti , Christian B. Andersen , Laura J. Grenville-Briggs
{"title":"The hunt for sustainable biocontrol of oomycete plant pathogens, a case study of Phytophthora infestans","authors":"Maryam Hashemi , Dania Tabet , Murilo Sandroni , Clara Benavent-Celma , Jenifer Seematti , Christian B. Andersen , Laura J. Grenville-Briggs","doi":"10.1016/j.fbr.2021.11.003","DOIUrl":"10.1016/j.fbr.2021.11.003","url":null,"abstract":"<div><p>Late blight caused by the oomycete <em>Phytophthora infestans</em> is considered to be one of the most severe diseases of potato and tomato worldwide. Whilst current synthetic fungicides are efficient at controlling this disease, they are an environmental and economic burden. In line with EU directives to reduce the use of synthetic pesticides and increase the use of sustainable alternative disease control strategies that can form part of integrated pest management systems, practical biological control solutions are urgently needed. Despite the fact that there has been a large body of scientific research into microorganisms with potential for the biological control of late blight disease, relatively few commercial biocontrol agents, licensed to control late blight, exist. Furthermore, the practical uptake of those in Europe is lower than might be expected, suggesting that such solutions are not yet feasible, or effective. Here we review the scientific literature, focusing on the most recent developments in the hunt for efficient and sustainable biological control of late blight disease. We discuss the progress in our mechanistic understanding of mycoparasite–prey interactions, in the context of late blight and the challenges and limitations to the use of such knowledge in practical disease control within a European context.</p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"40 ","pages":"Pages 53-69"},"PeriodicalIF":6.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1749461321000555/pdfft?md5=56c8be4b68ffcb5c19802a74681069d8&pid=1-s2.0-S1749461321000555-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45662413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Thomas Becking , Andrei Kiselev , Valentina Rossi , David Street-Jones , Frédéric Grandjean , Elodie Gaulin
{"title":"Pathogenicity of animal and plant parasitic Aphanomyces spp and their economic impact on aquaculture and agriculture","authors":"Thomas Becking , Andrei Kiselev , Valentina Rossi , David Street-Jones , Frédéric Grandjean , Elodie Gaulin","doi":"10.1016/j.fbr.2021.08.001","DOIUrl":"10.1016/j.fbr.2021.08.001","url":null,"abstract":"<div><p>Parasitic <span><em>Aphanomyces</em></span> species are a global threat to agri- and aquaculture, causing multimillion USD damage every year. Via the global trade, <em>Aphanomyces</em><span> has spread across all continents with exception of South America and Antarctica, and has become a major problem in pea, sugar beet, fish and crayfish production. The widespread </span><em>A. euteiches</em> and <em>A</em>. <em>cochlioides</em> induce root rot diseases in leguminous species and sugar beet respectively. The fish pathogen <em>A. invadans</em><span> is the causative agent of Epizootic Ulcerative Syndrome in various fish species whilst </span><em>A. astaci</em> infects crayfishes causing crayfish plague. <em>Aphanomyces</em> have developed an efficient transmission and infection mechanism which allows a rapid colonization and disruption of the host's infected tissues. This review presents an overview on the current research on <em>Aphanomyces</em> genus. We summarise the latest research efforts on four pathogenic <em>Aphanomyces</em><span> species, shedding light on the biology of these microorganisms, the pathogenicity factors of these parasites, the diseases which they cause, their distribution and finally the strategies to control the diseases.</span></p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"40 ","pages":"Pages 1-18"},"PeriodicalIF":6.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.fbr.2021.08.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42332991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"LysM proteins in mammalian fungal pathogens","authors":"José A. Oguiza","doi":"10.1016/j.fbr.2022.02.001","DOIUrl":"10.1016/j.fbr.2022.02.001","url":null,"abstract":"<div><p>The LysM domain is a highly conserved carbohydrate-binding module that recognizes polysaccharides containing N-acetylglucosamine residues. LysM domains are found in a wide variety of extracellular proteins and receptors from viruses, bacteria, fungi, plants and animals. LysM proteins are also present in many species of mammalian fungal pathogens, although a limited number of studies have focused on the expression and determination of their putative roles in the infection process. This review summarizes the current knowledge and recent studies on LysM proteins in the main morphological groups of fungal pathogens that cause infections in humans and other mammals. Recent advances towards understanding the biological functions of LysM proteins in infections of mammalian hosts and their use as potential targets in antifungal strategies are also discussed.</p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"40 ","pages":"Pages 114-122"},"PeriodicalIF":6.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1749461322000082/pdfft?md5=3485aa362a6567b92b883c6099b71075&pid=1-s2.0-S1749461322000082-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48467450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Clara Benavent-Celma , Noelia López-García , Tahmina Ruba , Magdalena E. Ściślak , David Street-Jones , Pieter van West , Stephen Woodward , Johanna Witzell
{"title":"Current practices and emerging possibilities for reducing the spread of oomycete pathogens in terrestrial and aquatic production systems in the European Union","authors":"Clara Benavent-Celma , Noelia López-García , Tahmina Ruba , Magdalena E. Ściślak , David Street-Jones , Pieter van West , Stephen Woodward , Johanna Witzell","doi":"10.1016/j.fbr.2021.10.001","DOIUrl":"10.1016/j.fbr.2021.10.001","url":null,"abstract":"<div><p>Diseases caused by oomycete<span><span> pathogens are a global threat to forestry<span>, agriculture and aquaculture. Because of their complex life cycles, characterised by dormant resting structures that enable their survival for years under hostile environmental conditions, reducing the spread of oomycetes is a challenging task. In this review, we present an overview of this challenge, starting from the need to understand the natural and anthropogenic dispersal pathways of these pathogens. Focusing on the European Union, we explore current legislation that forms a </span></span>backbone<span> for biosecurity<span> protocols against the spread of oomycetes through trade and transport. We discuss the options for prevention, containment and long-term management of oomycetes in different production settings, emphasising the importance of prevention as the most cost-efficient strategy to reduce the spread of these pathogens. Finally, we highlight some of the new and emerging technologies and strategies as potential tools in the integrated pest management of animal and plant diseases caused by oomycetes. We emphasise the urgency of actions to halt the global spread of these pathogens.</span></span></span></p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"40 ","pages":"Pages 19-36"},"PeriodicalIF":6.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41727391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bikal Ghimire , Marcia Saraiva , Christian B. Andersen , Anupam Gogoi , Mona Saleh , Nicola Zic , Pieter van West , May B. Brurberg
{"title":"Transformation systems, gene silencing and gene editing technologies in oomycetes","authors":"Bikal Ghimire , Marcia Saraiva , Christian B. Andersen , Anupam Gogoi , Mona Saleh , Nicola Zic , Pieter van West , May B. Brurberg","doi":"10.1016/j.fbr.2021.11.001","DOIUrl":"10.1016/j.fbr.2021.11.001","url":null,"abstract":"<div><p>Oomycetes are spore-forming eukaryotic microbes responsible for infections in animal and plant species worldwide, posing a threat to natural ecosystems, biodiversity and food security. Genomics and transcriptomics approaches, together with host interaction studies, give promising results towards better understanding of the infection mechanisms in oomycetes and their general biology. Significant development and progress in oomycetes genomic studies have been achieved over the past decades but further understanding of molecular processes, gene regulations and infection mechanisms are still needed. The use of molecular tools such as CRISPR/Cas and RNAi helped elucidate some of the molecular processes involved in host invasion and infection both in plant and animal pathogenic oomycetes. These methods provide an opportunity for accurate and detailed functional analysis involving various fields of studies such as genomics, epigenomics, proteomics, and interactomics. Functional gene characterisation is essential for filling the knowledge gaps in dynamic biological processes. However, every method has both advantages and limitations that should be considered before choosing the best method for investigating a particular research question. Here we review transformation systems, gene silencing and gene editing techniques in oomycetes, how they function, in which species and what are their main advantages and disadvantages.</p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"40 ","pages":"Pages 37-52"},"PeriodicalIF":6.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1749461321000531/pdfft?md5=48bca88a19c9c32d477434b3cd960243&pid=1-s2.0-S1749461321000531-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44873284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Isabel Vicente , Riccardo Baroncelli , Rosa Hermosa , Enrique Monte , Giovanni Vannacci , Sabrina Sarrocco
{"title":"Role and genetic basis of specialised secondary metabolites in Trichoderma ecophysiology","authors":"Isabel Vicente , Riccardo Baroncelli , Rosa Hermosa , Enrique Monte , Giovanni Vannacci , Sabrina Sarrocco","doi":"10.1016/j.fbr.2021.12.004","DOIUrl":"https://doi.org/10.1016/j.fbr.2021.12.004","url":null,"abstract":"<div><p>Species of fungal genus <span><em>Trichoderma</em></span><span> are characterized by a versatile lifestyle, high adaptability to the changing environmental conditions and the ability to establish sophisticated interactions with other organisms. Due to their ability to antagonize plant pathogens and to elicit the plant defence responses against biotic/abiotic stresses, </span><em>Trichoderma</em><span> spp. are commonly used as commercially biopesticides<span> and biofertilizers. The </span></span><em>Trichoderma</em><span> success in the rhizosphere<span> is supported by a wide arsenal of specialised metabolites (SMs) providing morphological and physiological autoregulation, self-protection and facilitating fungal communication. This review aims to explore the roles of SMs in the biology of fungi, with special emphasis on the genus </span></span><em>Trichoderma</em><span> and on how divergence in the SMs genetic structure determine </span><em>Trichoderma</em> lifestyles. <em>Trichoderma</em><span> genomes are endowed with a high number of SMs biosynthetic genes, and understanding the genetic basis of their biosynthesis is crucial for determining the role of these metabolites in </span><em>Trichoderma</em><span><span> ecophysiology and for expanding their application in </span>crop protection. Recent advances on the characterization of the </span><em>Trichoderma</em> SMs genetic inventory driven by computational biology are discussed.</p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"39 ","pages":"Pages 83-99"},"PeriodicalIF":6.0,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72049254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Cellular heterogeneity: yeast-side story","authors":"Sandrine Pinheiro, Shashank Pandey, Serge Pelet","doi":"10.1016/j.fbr.2021.11.005","DOIUrl":"10.1016/j.fbr.2021.11.005","url":null,"abstract":"<div><p>A major challenge for cells lies in their ability to detect, respond and adapt to changing environments that may threaten their survival. Among the numerous evolutionary strategies, cell-to-cell heterogeneity allows the emergence of different phenotypes within a population. This variability in cellular behaviors can be essential for a small fraction of cells to adapt and survive in various environments. Analyses at the single-cell level have allowed to highlight the great variability that is present between cells within an isogenic population. Numerous molecular mechanisms have been uncovered, allowing to understand the emergence and the role of cellular heterogeneity. These attempts at identifying the source of cellular noise have also provided clues for strategies needed to control heterogeneity. In this review, <em>S. cerevisiae</em> is used as an example to illustrate the different factors leading to cell heterogeneity, ranging from intracellular processes to environmental constraints. In addition, some recent strategies developed to modulate cell-to-cell variability are discussed.</p></div>","PeriodicalId":12563,"journal":{"name":"Fungal Biology Reviews","volume":"39 ","pages":"Pages 34-45"},"PeriodicalIF":6.0,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1749461321000579/pdfft?md5=f94b3732ec697cf3c655927b333df845&pid=1-s2.0-S1749461321000579-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54682785","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}