Flavio Luis de Oliveira, Raquel Silva, J. Morais, Pedro Cruz, V. Vasconcelos
: Cyanobacteria are microorganisms that have remarkable adaptability and can inhabit various types of aquatic and terrestrial ecosystems worldwide, including extreme environments. This group of organisms is considered a rich source of secondary metabolites with potential biotech-nological applications and has the capability to produce some potent cyanotoxins that can induce consequences to human health. The Blue Biotechnology and Ecotoxicology Culture Collection (LEGE-CC) is a biological resource center located at the Interdisciplinary Centre of Marine and Environmental Research (CIIMAR), comprising more than 1200 different cyanobacterial and microalgae strains. Until now, 36 strains have been reported as producers of cyanotoxins distributed within different cyanobacterial orders. Recently, LEGE-CC has increased in numbers due to the isolation effort that has been made. In this work, a screening of more than 200 cyanobacterial isolates from subaerial and freshwater environments targeted the genes involved in the biosynthesis of cyanotoxins. As expected, genes involved in cylindrospermopsin, saxitoxin, anatoxin and microcystin production were detected by molecular biology tools. The strains where the genes were detected were grown and sent to liquid chromatography–mass spectrometry (LC-MS) to confirm the production of cyanotoxins. As a culture collection, the screening of cyanotoxins is an essential aspect of cyanobacterial research and provides a comprehensive idea of the production of these toxins for future works.
{"title":"Cyanobacterial Toxins—An Update of Toxins from Blue Biotechnology and Ecotoxicology Culture Collection (LEGE-CC)","authors":"Flavio Luis de Oliveira, Raquel Silva, J. Morais, Pedro Cruz, V. Vasconcelos","doi":"10.3390/blsf2022014039","DOIUrl":"https://doi.org/10.3390/blsf2022014039","url":null,"abstract":": Cyanobacteria are microorganisms that have remarkable adaptability and can inhabit various types of aquatic and terrestrial ecosystems worldwide, including extreme environments. This group of organisms is considered a rich source of secondary metabolites with potential biotech-nological applications and has the capability to produce some potent cyanotoxins that can induce consequences to human health. The Blue Biotechnology and Ecotoxicology Culture Collection (LEGE-CC) is a biological resource center located at the Interdisciplinary Centre of Marine and Environmental Research (CIIMAR), comprising more than 1200 different cyanobacterial and microalgae strains. Until now, 36 strains have been reported as producers of cyanotoxins distributed within different cyanobacterial orders. Recently, LEGE-CC has increased in numbers due to the isolation effort that has been made. In this work, a screening of more than 200 cyanobacterial isolates from subaerial and freshwater environments targeted the genes involved in the biosynthesis of cyanotoxins. As expected, genes involved in cylindrospermopsin, saxitoxin, anatoxin and microcystin production were detected by molecular biology tools. The strains where the genes were detected were grown and sent to liquid chromatography–mass spectrometry (LC-MS) to confirm the production of cyanotoxins. As a culture collection, the screening of cyanotoxins is an essential aspect of cyanobacterial research and provides a comprehensive idea of the production of these toxins for future works.","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133265458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
S. Pereira, Leonor Ferreira, Catarina I. Gonçalves, V. Vasconcelos, Mariana A. Reis, J. Almeida
: The usage of paints and coatings with toxic components for the mitigation of marine biofouling in submerged surfaces continues to cause economic, environmental and human health-related problems worldwide. Natural products have the potential to provide solutions for antifouling applications that are effective and ecologically compatible. The diversity of the secondary metabolites that are produced by cyanobacteria make these organisms a promising source of bioactive compounds, especially when antifouling activity has already been documented. The purpose of this study was to explore the metabolic diversity of a range of cyanobacterial strains from the Blue Biotechnology and Ecotoxicology Culture Collection (LEGE-CC) in search of eco-friendly bioactive compounds for antifouling purposes. A library of fractions, derived from methanolic extracts, belonging to different cyanobacterial strains, was tested towards a prominent macrofouling organism settlement ( Mytilus galloprovincialis larvae). Promising fractions were submitted to a bioassay guided sub-fractioning that led to the isolation of two compounds. Their structure elucidation was determined by 1D and 2D nuclear magnetic resonance and by mass spectrometry. Anti-settlement effectiveness was assessed through an EC50 bioassay with mussel larvae, as well as antifouling bioactivity towards the growth of five marine biofilm-forming bacteria. The results showed bioactivity against the mussel larvae settlement and low toxicity, but no bacterial growth inhibition was found for the nucleosides (<10% of inhibition). Moreover, general ecotoxicity to the marine environment was evaluated, and the compounds also presented no toxicity against Artemia salina , proving them to be ecologically compatible. These promising results confirm the inherent potential of cyanobacteria to provide more sustainable antifouling ingredients to be incorporated in marine coatings.
{"title":"Cyanobacteria as a Source of New Antifouling Sustainable Solutions","authors":"S. Pereira, Leonor Ferreira, Catarina I. Gonçalves, V. Vasconcelos, Mariana A. Reis, J. Almeida","doi":"10.3390/blsf2022014037","DOIUrl":"https://doi.org/10.3390/blsf2022014037","url":null,"abstract":": The usage of paints and coatings with toxic components for the mitigation of marine biofouling in submerged surfaces continues to cause economic, environmental and human health-related problems worldwide. Natural products have the potential to provide solutions for antifouling applications that are effective and ecologically compatible. The diversity of the secondary metabolites that are produced by cyanobacteria make these organisms a promising source of bioactive compounds, especially when antifouling activity has already been documented. The purpose of this study was to explore the metabolic diversity of a range of cyanobacterial strains from the Blue Biotechnology and Ecotoxicology Culture Collection (LEGE-CC) in search of eco-friendly bioactive compounds for antifouling purposes. A library of fractions, derived from methanolic extracts, belonging to different cyanobacterial strains, was tested towards a prominent macrofouling organism settlement ( Mytilus galloprovincialis larvae). Promising fractions were submitted to a bioassay guided sub-fractioning that led to the isolation of two compounds. Their structure elucidation was determined by 1D and 2D nuclear magnetic resonance and by mass spectrometry. Anti-settlement effectiveness was assessed through an EC50 bioassay with mussel larvae, as well as antifouling bioactivity towards the growth of five marine biofilm-forming bacteria. The results showed bioactivity against the mussel larvae settlement and low toxicity, but no bacterial growth inhibition was found for the nucleosides (<10% of inhibition). Moreover, general ecotoxicity to the marine environment was evaluated, and the compounds also presented no toxicity against Artemia salina , proving them to be ecologically compatible. These promising results confirm the inherent potential of cyanobacteria to provide more sustainable antifouling ingredients to be incorporated in marine coatings.","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125920098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
N. G. Silva, M. Preto, V. Vasconcelos, R. Urbatzka
: Environmental and lifestyle adaptations over the last decades have contributed to enhanced man ´ s longevity, however it also paved the ground for different diseases to unfold. Today one of the main public health challenges is obesity and its related metabolic dysfunctions. In recent years, the pharmacological options are reported as being insufficient, therefore, the discovery and pharmacological development of new therapeutic approaches are required to overcome this epidemic. Vitamin K are a fat-soluble family of compounds implicated in a number of essential biological functions. Vitamin K1 and K2 are two naturally occurring compounds, while vitamin K3 is the most common synthetic form. A bioactivity-guided approach was used with the Nile red fat metabolism assay in zebrafish larvae to successfully isolate an analog of vitamin K1 for the first time from Tychonema sp. LEGE 07196. The structure confirmation was based on NMR spectroscopy and mass spectrometry. This analog was first isolated in 1965 from the cyanobacteria Synechococcus elongatus , but little is known on its bioactivity. The anti-obesity effects of all vitamin K forms, including the K1-analog, were studied. The compounds did not cause any general toxicity or malformations and showed significant neutral lipid-reducing activity after 48 h of exposure. The different vitamin forms displayed different levels of activity which shows the importance of the naphthoquinone ring, as well as the impact of the saturation and polarity of the aliphatic side-chain for the structure–activity relationship. Metabolomics approaches were employed to establish the distinct bioavailability and biotransformation of the different forms of vitamins in the organisms. The metabolite profiling was achieved using different databases and MetaboAnalyst was used for associated analysis. The organism has a clear preference to transform the various vitamins into K1 and K1-analog forms, regardless of the supplementation, and each exposed vitamin significatively altered the expression of different metabolites indicating that different metabolic targets are involved. This work is ongoing, and the final aim is to understand the effects of vitamin K family on obesity and related co-morbidities, which could lay the ground to develop a future nutraceutical with lipid reduction activity. conceptualization,
{"title":"The Role of Vitamin K Family in Obesity","authors":"N. G. Silva, M. Preto, V. Vasconcelos, R. Urbatzka","doi":"10.3390/blsf2022014036","DOIUrl":"https://doi.org/10.3390/blsf2022014036","url":null,"abstract":": Environmental and lifestyle adaptations over the last decades have contributed to enhanced man ´ s longevity, however it also paved the ground for different diseases to unfold. Today one of the main public health challenges is obesity and its related metabolic dysfunctions. In recent years, the pharmacological options are reported as being insufficient, therefore, the discovery and pharmacological development of new therapeutic approaches are required to overcome this epidemic. Vitamin K are a fat-soluble family of compounds implicated in a number of essential biological functions. Vitamin K1 and K2 are two naturally occurring compounds, while vitamin K3 is the most common synthetic form. A bioactivity-guided approach was used with the Nile red fat metabolism assay in zebrafish larvae to successfully isolate an analog of vitamin K1 for the first time from Tychonema sp. LEGE 07196. The structure confirmation was based on NMR spectroscopy and mass spectrometry. This analog was first isolated in 1965 from the cyanobacteria Synechococcus elongatus , but little is known on its bioactivity. The anti-obesity effects of all vitamin K forms, including the K1-analog, were studied. The compounds did not cause any general toxicity or malformations and showed significant neutral lipid-reducing activity after 48 h of exposure. The different vitamin forms displayed different levels of activity which shows the importance of the naphthoquinone ring, as well as the impact of the saturation and polarity of the aliphatic side-chain for the structure–activity relationship. Metabolomics approaches were employed to establish the distinct bioavailability and biotransformation of the different forms of vitamins in the organisms. The metabolite profiling was achieved using different databases and MetaboAnalyst was used for associated analysis. The organism has a clear preference to transform the various vitamins into K1 and K1-analog forms, regardless of the supplementation, and each exposed vitamin significatively altered the expression of different metabolites indicating that different metabolic targets are involved. This work is ongoing, and the final aim is to understand the effects of vitamin K family on obesity and related co-morbidities, which could lay the ground to develop a future nutraceutical with lipid reduction activity. conceptualization,","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129656980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Claudia Fournier, A. Quesada, A. Justel, A. Monteoliva, Jordi Cirera, C. Sola, A. Munné, Juan C. García, José Javier Rodríguez, S. Cirés
: Cyanobacterial harmful algal blooms (CyanoHABs) are expanding globally, representing a major risk for lakes and reservoirs due to their toxicity and economic impacts. Therefore, antic-ipating their occurrence and understanding the main factors related to CyanoHABs are critical to improve decision-making processes and water resource management. In this context, we present two modelling options for the analysis and prediction of cyanoHABs in two drinking water reservoirs from Spain. This case represents a unique opportunity to combine efforts from different academic disciplines (i.e., aquatic ecology and data science), environmental companies, and public water managers to address this increasingly severe issue. Susqueda (Ter basin, Catalonia) is a eutrophic, large and deep reservoir (Z max = 110 m) where monitoring efforts in recent years have focused on a monthly measurement in more than 30 physico-chemical, hydrological, meteorological and biological parameters, some of them involving expert intervention and costly efforts that could not be held at a higher temporary frequency. Cuerda del Pozo (Duero basin, Castilla y Le ó n) is a deep reservoir (Z max = 30 m) where monitoring efforts have focused on daily data collection through probes mounted in automatic profilers. This strategy allowed a higher monitoring frequency for fewer parameters and a narrower time span. In both cases, the parameter chosen as a proxy of cyanobacterial proliferation (output of models) is fluorometric measurements of chlorophyll-a and phycocyanin. The results of our machine-learning-based analyses suggest that the selected modelling path mainly depends on two aspects: (1) the time span where data are collected, and (2) the frequency and type of data measured (i.e., one discrete measurement at the surface vs. many measurements along the water column). Thus, a Susqueda dataset analysis led to more interpretative results, al-lowing for a better understanding of the system and the main factors related to CyanoHABs with limited predictive capacity. Meanwhile, the Cuerda del Pozo dataset is treated as a time series where autoregressive forecasting techniques, combined with information of exogenous parameters, are applied to foresee cyanobacterial blooms before they occur, losing part of the interpretability in the process. The results from this work are expected to provide an effective tool to boost smart and goal-orientated sampling planning, while improving data-driven decision-making processes essential for the water management of cyanobacterial blooms.
蓝藻有害藻华(CyanoHABs)正在全球范围内扩大，由于其毒性和经济影响，对湖泊和水库构成了重大风险。因此，预测其发生并了解与蓝藻有害藻华有关的主要因素对改善决策过程和水资源管理至关重要。在这种情况下，我们提出了两种建模方案，用于分析和预测西班牙两个饮用水水库中的蓝藻有害藻华。这个案例提供了一个独特的机会，可以将不同学科(即水生生态学和数据科学)、环境公司和公共水管理人员的努力结合起来，解决这一日益严重的问题。Susqueda (Ter basin, Catalonia)是一个富营养化、大而深的水库(zmax = 110 m)，近年来的监测工作主要集中在每月测量30多种物理化学、水文、气象和生物参数，其中一些涉及专家干预和昂贵的努力，无法以更高的临时频率进行。Cuerda del Pozo (Castilla y Le Duero basin， ó n)是一个深层油藏(最大水深30米)，监测工作主要集中在通过安装在自动剖面仪上的探针收集日常数据。该策略允许对更少的参数和更短的时间跨度进行更高的监测频率。在这两种情况下，选择作为蓝藻增殖代理的参数(模型的输出)是叶绿素-a和藻蓝蛋白的荧光测量。我们基于机器学习的分析结果表明，所选择的建模路径主要取决于两个方面:(1)收集数据的时间跨度，(2)测量数据的频率和类型(即，在地表进行一次离散测量与沿着水柱进行多次测量)。因此，通过对Susqueda数据集的分析，可以获得更具解释性的结果，从而更好地了解该系统以及与蓝藻藻华相关的主要因素，但预测能力有限。同时，Cuerda del Pozo数据集被视为一个时间序列，其中自回归预测技术结合外源参数信息，应用于在蓝藻繁殖发生之前预测，在此过程中失去了部分可解释性。这项工作的结果有望提供一个有效的工具，以促进智能和目标导向的采样计划，同时改善数据驱动的决策过程，对蓝藻华的水管理至关重要。
{"title":"Machine Learning Approach towards the Early Warning of Cyanobacterial Blooms in Drinking Water Reservoirs","authors":"Claudia Fournier, A. Quesada, A. Justel, A. Monteoliva, Jordi Cirera, C. Sola, A. Munné, Juan C. García, José Javier Rodríguez, S. Cirés","doi":"10.3390/blsf2022014038","DOIUrl":"https://doi.org/10.3390/blsf2022014038","url":null,"abstract":": Cyanobacterial harmful algal blooms (CyanoHABs) are expanding globally, representing a major risk for lakes and reservoirs due to their toxicity and economic impacts. Therefore, antic-ipating their occurrence and understanding the main factors related to CyanoHABs are critical to improve decision-making processes and water resource management. In this context, we present two modelling options for the analysis and prediction of cyanoHABs in two drinking water reservoirs from Spain. This case represents a unique opportunity to combine efforts from different academic disciplines (i.e., aquatic ecology and data science), environmental companies, and public water managers to address this increasingly severe issue. Susqueda (Ter basin, Catalonia) is a eutrophic, large and deep reservoir (Z max = 110 m) where monitoring efforts in recent years have focused on a monthly measurement in more than 30 physico-chemical, hydrological, meteorological and biological parameters, some of them involving expert intervention and costly efforts that could not be held at a higher temporary frequency. Cuerda del Pozo (Duero basin, Castilla y Le ó n) is a deep reservoir (Z max = 30 m) where monitoring efforts have focused on daily data collection through probes mounted in automatic profilers. This strategy allowed a higher monitoring frequency for fewer parameters and a narrower time span. In both cases, the parameter chosen as a proxy of cyanobacterial proliferation (output of models) is fluorometric measurements of chlorophyll-a and phycocyanin. The results of our machine-learning-based analyses suggest that the selected modelling path mainly depends on two aspects: (1) the time span where data are collected, and (2) the frequency and type of data measured (i.e., one discrete measurement at the surface vs. many measurements along the water column). Thus, a Susqueda dataset analysis led to more interpretative results, al-lowing for a better understanding of the system and the main factors related to CyanoHABs with limited predictive capacity. Meanwhile, the Cuerda del Pozo dataset is treated as a time series where autoregressive forecasting techniques, combined with information of exogenous parameters, are applied to foresee cyanobacterial blooms before they occur, losing part of the interpretability in the process. The results from this work are expected to provide an effective tool to boost smart and goal-orientated sampling planning, while improving data-driven decision-making processes essential for the water management of cyanobacterial blooms.","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129299144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
E. Santos, A. Massa, J. Azevedo, Diogo Martins, Mariana Reimão, Vítor Vasconcelos, A. Campos, M. Freitas
: The use of cyanobacterial biomass as a biofertilizer is promising in terms of sustainable agri-culture. Nevertheless, cyanobacteria can be considered a threat to human and environmental health due to the potential presence of cyanotoxins, since some studies report that the use of contaminated water for agricultural irrigation can impair plant growth and lead to contamination of food products. Interestingly, at environmentally relevant concentrations, cylindrospermopsin (CYN) seems to cause no deleterious effects in plants, and it might even promote their yield. However, studies assessing CYN accumulation in the edible tissues at environmental concentrations are lacking. The objective of this work was to evaluate the effects of cyanobacterial biomass CYN producing or non-producing on lettuce plant growth, and that of CYN accumulation in edible tissues. This study consisted of growing lettuce plants, under controlled conditions, for 25 days in soil (1) with no extra nutrient addition (control) and supplementation with (2) cyanobacterial biomass that did not produce CYN, (3) cyanobacterial biomass that produced CYN (~10 µ g of dissolved CYN), and (4) cyanobacterial biomass that produced CYN, treated by boiling for 5 min (~25 µ g of dissolved CYN). At the end of the exposure, lettuce growth was assessed, as well as CYN accumulation in tissues and soil. The results showed that leaf growth was significantly increased ( p < 0.05) in lettuce plants supplemented with cyanobacterial biomass, especially at condition (3), which was five-fold higher compared with the control group. Regarding CYN accumulation, for conditions (3) and (4), the toxin was detected in the tissues of plants, as well as in soil at the following decreasing order of concentrations: soil > roots > leaves. Interestingly, the concentration determined in lettuce leaves in condition (4) was three-fold lower when compared with the condition (3). Nevertheless, for both conditions, although CYN has been detected in lettuce leaves, the concentration in the edible part did not exceed the proposed provi-sional tolerable daily intake (TDI) of 0.03 µ g/kg/BW. In conclusion, these results suggest that the use of cyanobacterial biomass as lettuce biofertilizer, even containing CYN at environmentally relevant concentrations, can positively influence plant growth and development without compromising the safety of edible tissues. Contributions: Conceptualization, methodology, É D.M., software, É A.M., J.A. A.C.; validation, É A.M., J.A., M.F. and A.C.; formal analysis, É .S., A.M. and J.A.; investigation, É .S., A.M., J.A., M.F. and A.C.; resources, É .S., A.M., J.A., D.M., M.R., M.F., A.C. and V.V.; data curation, É .S., A.M. and J.A.; writing—original draft preparation, É A.M. and M.F.; writing—review and editing, É A.M., A.C., M.F. and V.V.; supervision, M.F. and A.C.; project administration, A.C. and V.V.; funding acquisition, A.C. and V.V.
{"title":"Cyanobacterial Biomass Used as Biofertilizer in Lettuce Plants: Effects on Growth and Cyanotoxin Accumulation","authors":"E. Santos, A. Massa, J. Azevedo, Diogo Martins, Mariana Reimão, Vítor Vasconcelos, A. Campos, M. Freitas","doi":"10.3390/blsf2022014035","DOIUrl":"https://doi.org/10.3390/blsf2022014035","url":null,"abstract":": The use of cyanobacterial biomass as a biofertilizer is promising in terms of sustainable agri-culture. Nevertheless, cyanobacteria can be considered a threat to human and environmental health due to the potential presence of cyanotoxins, since some studies report that the use of contaminated water for agricultural irrigation can impair plant growth and lead to contamination of food products. Interestingly, at environmentally relevant concentrations, cylindrospermopsin (CYN) seems to cause no deleterious effects in plants, and it might even promote their yield. However, studies assessing CYN accumulation in the edible tissues at environmental concentrations are lacking. The objective of this work was to evaluate the effects of cyanobacterial biomass CYN producing or non-producing on lettuce plant growth, and that of CYN accumulation in edible tissues. This study consisted of growing lettuce plants, under controlled conditions, for 25 days in soil (1) with no extra nutrient addition (control) and supplementation with (2) cyanobacterial biomass that did not produce CYN, (3) cyanobacterial biomass that produced CYN (~10 µ g of dissolved CYN), and (4) cyanobacterial biomass that produced CYN, treated by boiling for 5 min (~25 µ g of dissolved CYN). At the end of the exposure, lettuce growth was assessed, as well as CYN accumulation in tissues and soil. The results showed that leaf growth was significantly increased ( p < 0.05) in lettuce plants supplemented with cyanobacterial biomass, especially at condition (3), which was five-fold higher compared with the control group. Regarding CYN accumulation, for conditions (3) and (4), the toxin was detected in the tissues of plants, as well as in soil at the following decreasing order of concentrations: soil > roots > leaves. Interestingly, the concentration determined in lettuce leaves in condition (4) was three-fold lower when compared with the condition (3). Nevertheless, for both conditions, although CYN has been detected in lettuce leaves, the concentration in the edible part did not exceed the proposed provi-sional tolerable daily intake (TDI) of 0.03 µ g/kg/BW. In conclusion, these results suggest that the use of cyanobacterial biomass as lettuce biofertilizer, even containing CYN at environmentally relevant concentrations, can positively influence plant growth and development without compromising the safety of edible tissues. Contributions: Conceptualization, methodology, É D.M., software, É A.M., J.A. A.C.; validation, É A.M., J.A., M.F. and A.C.; formal analysis, É .S., A.M. and J.A.; investigation, É .S., A.M., J.A., M.F. and A.C.; resources, É .S., A.M., J.A., D.M., M.R., M.F., A.C. and V.V.; data curation, É .S., A.M. and J.A.; writing—original draft preparation, É A.M. and M.F.; writing—review and editing, É A.M., A.C., M.F. and V.V.; supervision, M.F. and A.C.; project administration, A.C. and V.V.; funding acquisition, A.C. and V.V.","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129515663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Joana Andrade, Catarina Silva, Luís Vieira, M. Pinto, J. P. Gomes, E. Valério
: Cyanobacteria are a ubiquitous and diverse group of phototrophic prokaryotes, which mainly inhabit aquatic ecosystems. In certain optimal environmental conditions, there may be a rapid increase in cyanobacteria populations, leading to the formation of blooms, which are frequently associated with the presence of cyanotoxins. Microcystins (MCs) are the most frequent hepatotoxin produced by cyanobacteria. Scarce previous studies have shown that the growth of aquatic heterotrophic bacteria, which co-occur with cyanobacteria, may not be affected by the presence of MCs, or may present a reduction, never being totally inhibited by their presence. In this study, we examined the effects of three microcystin variants (MCLR, MCRR and MCYR) on a set of heterotrophic aquatic bacteria living in the same ecosystem as cyanobacteria. In particular, the impact of microcystins on the growth of heterotrophic bacteria was tested, and a PCR screening for the presence of microcystin-degrading genes ( mlr ) was performed. The growth assays supported the hypothesis from previous studies, where most heterotrophic bacteria were only slightly or not at all affected by exposure to MCs. Moreover, it seems that the behavior of the isolates when exposed to these cyanotoxins was strain specific. A new bacteria, mlr +, was identified, belonging to Flectobacillus sp. Furthermore, we decided to perform a genomic study of 14 isolates from a set of potentially interesting bacteria, including Flavobacterium spp. and Aeromonas spp., to search for xenobiotic-related genes that could be involved in MC degradation. The whole-genome sequencing analysis of these 14 isolates revealed that no COG genes (COG0625; COG0841; COG1566) were present; however, genes similar to CAAX genes were present in the Aeromonas spp. isolates analyzed. These results shed new light into alternative molecular mechanisms for microcystin degradation.
{"title":"Assessing the Reason Why Heterotrophic Bacteria Present in Aquatic Environments Are Not Affected by Microcystins and Unraveling Alternative Genes for Microcystin Degradation","authors":"Joana Andrade, Catarina Silva, Luís Vieira, M. Pinto, J. P. Gomes, E. Valério","doi":"10.3390/blsf2022014033","DOIUrl":"https://doi.org/10.3390/blsf2022014033","url":null,"abstract":": Cyanobacteria are a ubiquitous and diverse group of phototrophic prokaryotes, which mainly inhabit aquatic ecosystems. In certain optimal environmental conditions, there may be a rapid increase in cyanobacteria populations, leading to the formation of blooms, which are frequently associated with the presence of cyanotoxins. Microcystins (MCs) are the most frequent hepatotoxin produced by cyanobacteria. Scarce previous studies have shown that the growth of aquatic heterotrophic bacteria, which co-occur with cyanobacteria, may not be affected by the presence of MCs, or may present a reduction, never being totally inhibited by their presence. In this study, we examined the effects of three microcystin variants (MCLR, MCRR and MCYR) on a set of heterotrophic aquatic bacteria living in the same ecosystem as cyanobacteria. In particular, the impact of microcystins on the growth of heterotrophic bacteria was tested, and a PCR screening for the presence of microcystin-degrading genes ( mlr ) was performed. The growth assays supported the hypothesis from previous studies, where most heterotrophic bacteria were only slightly or not at all affected by exposure to MCs. Moreover, it seems that the behavior of the isolates when exposed to these cyanotoxins was strain specific. A new bacteria, mlr +, was identified, belonging to Flectobacillus sp. Furthermore, we decided to perform a genomic study of 14 isolates from a set of potentially interesting bacteria, including Flavobacterium spp. and Aeromonas spp., to search for xenobiotic-related genes that could be involved in MC degradation. The whole-genome sequencing analysis of these 14 isolates revealed that no COG genes (COG0625; COG0841; COG1566) were present; however, genes similar to CAAX genes were present in the Aeromonas spp. isolates analyzed. These results shed new light into alternative molecular mechanisms for microcystin degradation.","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131247862","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T. Rosado, Duarte Balata, R. de la Rivière, V. Manageiro, D. Carpinteiro, Luís Vieira, F. Pina-Martins, C. Menezes, O. Paulo, M. Caniça, E. Dias
: The aquatic environment is a reservoir of antibiotics, pathogenic and native microorganisms, and antibiotic resistance genes (ARGs), constituting a key aspect of the One Health approach. Thus, the problem of antibiotic resistance is no longer restricted to bacterial pathogens but is a consequence of the interplay between distinct clinical and environmental drivers. In the project “ExplorAR— Exploring the Aquatic Resistome”, we investigated the putative role of cyanobacteria (CB) in the freshwater resistome, using a multidisciplinary approach based on phenotypic/genotypic/genomic tools. In this presentation, we disclose the main results of the project: (i) the development of a microdilution-based antibiotic susceptibility assay for CB; (ii) the establishment of an antibiotic susceptibility profile of CB genus/species; (iii) the identification of CB strains with reduced susceptibility to multiple antibiotic classes; (iv) the identification of ARGs in CB genomes; (v) the characterization of the antibiotic resistance profile of CB-associated bacteria; (vi) mapping the occurrence of CB and ARGs in surface freshwater reservoirs by high-throughput sequencing approaches. Overall, ExplorAR strongly supports the hypothesis that CB are environmental players in the emergence and dissemination of ARGs in water environments, contributing to the problem, and challenges, of antibiotic resistance.
{"title":"The Role of Cyanobacteria in the Aquatic Resistome","authors":"T. Rosado, Duarte Balata, R. de la Rivière, V. Manageiro, D. Carpinteiro, Luís Vieira, F. Pina-Martins, C. Menezes, O. Paulo, M. Caniça, E. Dias","doi":"10.3390/blsf2022014032","DOIUrl":"https://doi.org/10.3390/blsf2022014032","url":null,"abstract":": The aquatic environment is a reservoir of antibiotics, pathogenic and native microorganisms, and antibiotic resistance genes (ARGs), constituting a key aspect of the One Health approach. Thus, the problem of antibiotic resistance is no longer restricted to bacterial pathogens but is a consequence of the interplay between distinct clinical and environmental drivers. In the project “ExplorAR— Exploring the Aquatic Resistome”, we investigated the putative role of cyanobacteria (CB) in the freshwater resistome, using a multidisciplinary approach based on phenotypic/genotypic/genomic tools. In this presentation, we disclose the main results of the project: (i) the development of a microdilution-based antibiotic susceptibility assay for CB; (ii) the establishment of an antibiotic susceptibility profile of CB genus/species; (iii) the identification of CB strains with reduced susceptibility to multiple antibiotic classes; (iv) the identification of ARGs in CB genomes; (v) the characterization of the antibiotic resistance profile of CB-associated bacteria; (vi) mapping the occurrence of CB and ARGs in surface freshwater reservoirs by high-throughput sequencing approaches. Overall, ExplorAR strongly supports the hypothesis that CB are environmental players in the emergence and dissemination of ARGs in water environments, contributing to the problem, and challenges, of antibiotic resistance.","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127684198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Paula Soms, Rebeca Pérez-González, X. Sòria-Perpinyà, J. Soria, E. Vicente
: Some cyanobacteria are considered beneficial to humans, while others are considered harmful due to their ability to synthesize and release cyanotoxins. Cyanobacterial blooms are proliferations of cyanobacteria due to the high concentration of nutrients such as nitrogen and phosphorus, as well as the intervention of other physicochemical parameters such as solar radiation and temperature, worsening the trophic state of the water body. Cyanobacterial bloom can produce cyanotoxins, which directly decrease water quality for human use. Since the 1980s, urban and agricultural intensification, together with the effects of climate change, produced the current poor trophic state of the Albufera of Valencia. It is a coastal lagoon located in the east of the Iberian Peninsula, which requires continuous monitoring in order to monitor its ecological quality. The objective of this study is to monitor the concentration of phycocyanin and its relationship with the density of cyanobacteria during the winter–spring of 2022. For this, physicochemical variables were measured, and phytoplankton communities were determined; finally, the presence of phycocyanin was measured in situ and by remote sensing using Sentinel-2 imagery. The lagoon presents a state of permanent turbidity with an average transparency of 30 cm according to the Secchi Disk. The results obtained show an increasing concentration of phycocyanin from winter to summer related to higher phosphorus concentrations, reaching values of 561.3 µ g/L for phycocyanin and 8.7 mg P/L of total phosphorus. According to the trophic state reference values of the Water Framework Directive, the concentration of total phosphorus is excessive, indicating a hypertrophic state. According to the World Health Organization, the concentration of phycocyanin found would indicate a state of high alert. Remote sensing serves as a suitable tool for monitoring and tracking the status of the lagoon.
{"title":"Monitoring Cyanobacteria and Phycocyanin: A Case Study in the Albufera of Valencia","authors":"Paula Soms, Rebeca Pérez-González, X. Sòria-Perpinyà, J. Soria, E. Vicente","doi":"10.3390/blsf2022014031","DOIUrl":"https://doi.org/10.3390/blsf2022014031","url":null,"abstract":": Some cyanobacteria are considered beneficial to humans, while others are considered harmful due to their ability to synthesize and release cyanotoxins. Cyanobacterial blooms are proliferations of cyanobacteria due to the high concentration of nutrients such as nitrogen and phosphorus, as well as the intervention of other physicochemical parameters such as solar radiation and temperature, worsening the trophic state of the water body. Cyanobacterial bloom can produce cyanotoxins, which directly decrease water quality for human use. Since the 1980s, urban and agricultural intensification, together with the effects of climate change, produced the current poor trophic state of the Albufera of Valencia. It is a coastal lagoon located in the east of the Iberian Peninsula, which requires continuous monitoring in order to monitor its ecological quality. The objective of this study is to monitor the concentration of phycocyanin and its relationship with the density of cyanobacteria during the winter–spring of 2022. For this, physicochemical variables were measured, and phytoplankton communities were determined; finally, the presence of phycocyanin was measured in situ and by remote sensing using Sentinel-2 imagery. The lagoon presents a state of permanent turbidity with an average transparency of 30 cm according to the Secchi Disk. The results obtained show an increasing concentration of phycocyanin from winter to summer related to higher phosphorus concentrations, reaching values of 561.3 µ g/L for phycocyanin and 8.7 mg P/L of total phosphorus. According to the trophic state reference values of the Water Framework Directive, the concentration of total phosphorus is excessive, indicating a hypertrophic state. According to the World Health Organization, the concentration of phycocyanin found would indicate a state of high alert. Remote sensing serves as a suitable tool for monitoring and tracking the status of the lagoon.","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114410284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
: Cyanotoxins are chemical pollutants produced and released by Cyanobacteria, the oldest living prokaryotes, and have well-established implications for human and aquatic biota health. Due to current global conditions, it has become essential to offer an overview of the dynamics of cyanotoxins occurrence in order to improve the prediction and control of our water resources. In Portugal, in recent years, the impacts on global warming have contributed to hot abnormal events and a rise in temperature. Between May and October in two consecutive years (2017–2018), seven freshwater ecosystems located in the north and central regions of Portugal were assessed for the occurrence of microcystins (mcyA), cylindrospermopsins (cyrC), anatoxin-a (anaC), and saxitoxins (sxtI) by applying genetics methods. Our findings illustrate that, in a year with two heat waves (2017), the occurrence of blooms increased, as did the number on cyanotoxins detected compared with the less warm year (2018). Despite this trend, two ecosystems escaped and maintained an elevated risk with regard to cyanotoxin detection. DNA sequencing revealed the presence of potentially toxic cyanobacteria in all sampled ecosystems. The data retrieved highlighted genotypes for all main cyanotoxins. Continuous monitoring efforts are demanded in Portugal for improving knowledge of the occurrence of cyanotoxins and for future national regulations. Preliminary data also revealed that the impact of global change on Portugal has improved cyanotoxin detection.
{"title":"Cyanotoxins Dynamics on Portuguese Freshwater Ecosystems on Current Global Changes","authors":"Cristiana Moreira, V. Vasconcelos, A. Antunes","doi":"10.3390/blsf2022014030","DOIUrl":"https://doi.org/10.3390/blsf2022014030","url":null,"abstract":": Cyanotoxins are chemical pollutants produced and released by Cyanobacteria, the oldest living prokaryotes, and have well-established implications for human and aquatic biota health. Due to current global conditions, it has become essential to offer an overview of the dynamics of cyanotoxins occurrence in order to improve the prediction and control of our water resources. In Portugal, in recent years, the impacts on global warming have contributed to hot abnormal events and a rise in temperature. Between May and October in two consecutive years (2017–2018), seven freshwater ecosystems located in the north and central regions of Portugal were assessed for the occurrence of microcystins (mcyA), cylindrospermopsins (cyrC), anatoxin-a (anaC), and saxitoxins (sxtI) by applying genetics methods. Our findings illustrate that, in a year with two heat waves (2017), the occurrence of blooms increased, as did the number on cyanotoxins detected compared with the less warm year (2018). Despite this trend, two ecosystems escaped and maintained an elevated risk with regard to cyanotoxin detection. DNA sequencing revealed the presence of potentially toxic cyanobacteria in all sampled ecosystems. The data retrieved highlighted genotypes for all main cyanotoxins. Continuous monitoring efforts are demanded in Portugal for improving knowledge of the occurrence of cyanotoxins and for future national regulations. Preliminary data also revealed that the impact of global change on Portugal has improved cyanotoxin detection.","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125130596","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Campos, M. Freitas, B. Oudra, Vítor Vasconcelos
: Water contaminated with microcystins (MCs) or other cyanotoxins is recurrently used in agriculture and for crop irrigation. Several deleterious effects of MCs in plants that may impair crop productivity, including a decrease in growth and tissue necrosis, as well as an inhibition of photosynthesis and metabolic changes, have been reported. Studies also revealed a significant accumulation of MCs in edible tissues and plant organs, which raise concerns related to food safety. The European project TOXICROP precisely tackles this environmental problem. The main aims of the project are to map agricultural risk areas of cyanotoxin occurrence, to assess the fate of cyanotoxins in crops, and evaluate the impacts of using low-quality water for crop irrigation. The project also develops research on water remediation, exploring nature-based technologies. Here, we review part of the research carried out in the project, concerning the toxicity of cyanotoxins in crops. The research from TOXICROP Consortium has revealed for instance that adult strawberry or faba bean plants are susceptible to moderate concentrations of MCs (10 to 20 µ g MCs/L). Furthermore, experiments with faba bean and common wheat grown in sterile (microorganism-free) and non-sterile (microorganism-rich) soil, watered with 100 µ g MCs/L, revealed that native rhizospheric microbiota play an important role in the mitigation of the phytotoxic impact of MCs on plant growth, reducing toxin accumulation in both soils and plant tissues. Our studies also revealed that leaf vegetables, such as lettuce and spinach, growing in hydroponics are more susceptible to MCs than to the toxin, cylindrospermopsin (CYN). The lowest toxin concentrations affecting spinach and lettuce growth were 5 + 5 and 25 + 25 µ g/L CYN/MC mixtures, respectively. The results also reveal that the accumulation of MCs and CYN in plants depends on the conditions in which plants grow and concentrations of toxins in the irrigation water. In some cases, MCs are accumulated in plant tissues and exceed the tolerable daily intake proposed by the World Health Organization. We highlight the importance and contributions of this research to the definition and implementation of regulatory limits for cyanotoxins in irrigation waters.
{"title":"Contributions of TOXICROP Project for the Assessment of the Impacts of Toxic Cyanobacteria in Agriculture","authors":"A. Campos, M. Freitas, B. Oudra, Vítor Vasconcelos","doi":"10.3390/blsf2022014027","DOIUrl":"https://doi.org/10.3390/blsf2022014027","url":null,"abstract":": Water contaminated with microcystins (MCs) or other cyanotoxins is recurrently used in agriculture and for crop irrigation. Several deleterious effects of MCs in plants that may impair crop productivity, including a decrease in growth and tissue necrosis, as well as an inhibition of photosynthesis and metabolic changes, have been reported. Studies also revealed a significant accumulation of MCs in edible tissues and plant organs, which raise concerns related to food safety. The European project TOXICROP precisely tackles this environmental problem. The main aims of the project are to map agricultural risk areas of cyanotoxin occurrence, to assess the fate of cyanotoxins in crops, and evaluate the impacts of using low-quality water for crop irrigation. The project also develops research on water remediation, exploring nature-based technologies. Here, we review part of the research carried out in the project, concerning the toxicity of cyanotoxins in crops. The research from TOXICROP Consortium has revealed for instance that adult strawberry or faba bean plants are susceptible to moderate concentrations of MCs (10 to 20 µ g MCs/L). Furthermore, experiments with faba bean and common wheat grown in sterile (microorganism-free) and non-sterile (microorganism-rich) soil, watered with 100 µ g MCs/L, revealed that native rhizospheric microbiota play an important role in the mitigation of the phytotoxic impact of MCs on plant growth, reducing toxin accumulation in both soils and plant tissues. Our studies also revealed that leaf vegetables, such as lettuce and spinach, growing in hydroponics are more susceptible to MCs than to the toxin, cylindrospermopsin (CYN). The lowest toxin concentrations affecting spinach and lettuce growth were 5 + 5 and 25 + 25 µ g/L CYN/MC mixtures, respectively. The results also reveal that the accumulation of MCs and CYN in plants depends on the conditions in which plants grow and concentrations of toxins in the irrigation water. In some cases, MCs are accumulated in plant tissues and exceed the tolerable daily intake proposed by the World Health Organization. We highlight the importance and contributions of this research to the definition and implementation of regulatory limits for cyanotoxins in irrigation waters.","PeriodicalId":198127,"journal":{"name":"The 7th Iberian Congress on Cyanotoxins/3rd Iberoamerican Congress on Cyanotoxins","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116295625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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