ALGADEC - Detection of toxic algae with a semi-automated nucleic acid biosensor - Bewerkingsoverzicht

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 *[[Real-time algae monitoring]]
 *[[Eutrophication in coastal environments]]
 ===External Links===

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 In the course of the ALGADEC-project it was possible to develop a semi-automatic nucleic acid biosensor for the detection of toxic algae. The functionality of the device, even in the hands of lay persons, was shown with laboratory algae cultures, field samples spiked with algae cultures and field samples with naturally occurring toxic algae. However, in the future, the system has to be calibrated and optimised in respect to sensitivity for the detection of the target organisms. The sensitivity of the device is a crucial issue and has to be adapted, to the reference values for toxic algae e.g. toxic ''Alexandrium sp.'' in sea water of around ~100 – 250 cells/liter.
 The original idea of the ALGADEC project was to develop a nucleic acid biosensor for the detection of toxic algae. But the technology suggests an adaptation e.g. to the monitoring of microalgae in general. Therefore, molecular probes will be developed for key species of the phytoplankton of the North Sea. Furthermore, we are currently working on the automation of all steps involved in the analysis of water samples. In the long term a fully automated nucleic acid biosensor will be available that could work on its own or be implemented to the [[Ships_of_opportunity_and_ferries_as_instrument_carriers|FerryBox-System]] in order to monitor microalgae autonomously at species level.
 ==References==

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 {{featured}}
 ===Introduction===
-Microalgae are the major producers of [[Biomass|biomass]] and organic compounds in the aquatic environment. Among the marine microalgae there are 97 toxic species (mainly dinoflagellates) known to have the potential to form "[[Harmful_algal_blooms|Harmful Algal Blooms]]", the so called HABs (Fig. 1). [[Image:ALGADEC_1.jpg|thumb|left|'''Figure 1''' Bloom of ''Noctiluca scintillas'' in October 2002, Leigh, New Zealand.]] In recent decades, the public health and economic impacts of toxic algae species appear to have increased in frequency, intensity and geographic distribution (Zingone and Enevoldsen, 2000<ref name="Z&E">Zingone, A. & Enevoldsen, H.O. (2000). The diversity of harmful algal blooms: a challenge for science and management. Ocean & coastal management, 43, 725-748.</ref>, Daranas et al., 2001<ref name="D">Daranas A. H., Norte M. and Fernandez, J. J. (2001). Toxic marine microalgae, Toxicon, 39, 1101-1132.</ref>, Hallegraeff, 2003<ref name="H">Hallegraeff G. M. (2003). Harmful algal blooms: a global overview. In G.M. Hallegraeff, D.M. Anderson & A.D. Cembella (Eds.), Manual on Harmful Marine Microalgae (pp. 25-49). United Nations Educational, Scientific and Cultural Organization.</ref>, Moestrup, 2004.<ref name="M"> Moestrup, O. (2004). IOC Taxonomic Reference list of Toxic Algae. In O. Moestrup (Ed.), IOC taxonomic reference list of toxic algae. Intergovernmental Oceanographic Commission of the UNESCO.</ref>). In order to minimise the damage to human health or living resources, such as shellfish and fish, as well as economic losses to fisherman, aquaculture and the tourist industry, efficient [[State of the Art Overview on Field Observation Techniques (Theme 9)|monitoring methods]] are required for monitoring potentially toxic algal species (identification and quantification) (Andersen et al., 2003.<ref name="A"> Andersen P., Enevoldsen H. and Anderson, D.M. (2003). Harmful algal monitoring programme and action plan design. In G.M. Hallegraeff, D.M. Anderson & A.D. Cembella (Eds.), Manual on Harmful Marine Microalgae (pp. 627-647). United Nations Educational, Scientific and Cultural Organization.</ref>).  [[Image:ALGADEC_3.jpg|thumb|right|'''Figure 2''' Schematic drawing of a sandwich hybridisation. The target organism is identified by binding of two species specific molecular probes to the ribosomal RNA (rRNA). One of the probes is immobilised on the surface of the sensorchip and the other is coupled to digoxigenin, which binds to an antibody enzyme-complex. The enzyme catalyses a redox-reaction that can be measured as an electrochemical signal.]] The identification of unicelluar algae with conventional methods like light microscopy requires a thorough taxonomic expertise and is time-consuming. It is also costly if larger numbers of samples need to be processed. In some cases toxic and non-toxic varieties (strains) belong to the same species. They are morphologically identical, and cannot be distinguished by conventional methods. Consequently, improved monitoring methods that allow rapid detection and counting of toxic algae are needed. In the past decade, a variety of molecular methods have been adapted for the detection of harmful algae. Most of these techniques focus on the genetic information in the DNA or RNA (both nucleic acids) of the organisms. However, most of these new techniques are lab-based and not suited to be carried out in the field.
+Microalgae are the major producers of [[Biomass|biomass]] and organic compounds in the aquatic environment. Among the marine microalgae there are 97 toxic species (mainly dinoflagellates) known to have the potential to form "[[Harmful_algal_blooms|Harmful Algal Blooms]]", the so called HABs (Fig. 1). [[Image:ALGADEC_1.jpg|thumb|left|'''Figure 1''' Bloom of ''Noctiluca scintillas'' in October 2002, Leigh, New Zealand.]] In recent decades, the public health and economic impacts of toxic algae species appear to have increased in frequency, intensity and geographic distribution (Zingone and Enevoldsen, 2000<ref name="Z&E">Zingone, A. & Enevoldsen, H.O. (2000). The diversity of harmful algal blooms: a challenge for science and management. Ocean & coastal management, 43, 725-748.</ref>, Daranas et al., 2001<ref name="D">Daranas A. H., Norte M. and Fernandez, J. J. (2001). Toxic marine microalgae, Toxicon, 39, 1101-1132.</ref>, Hallegraeff, 2003<ref name="H">Hallegraeff G. M. (2003). Harmful algal blooms: a global overview. In G.M. Hallegraeff, D.M. Anderson & A.D. Cembella (Eds.), Manual on Harmful Marine Microalgae (pp. 25-49). United Nations Educational, Scientific and Cultural Organization.</ref>, Moestrup, 2004.<ref name="M"> Moestrup, O. (2004). IOC Taxonomic Reference list of Toxic Algae. In O. Moestrup (Ed.), IOC taxonomic reference list of toxic algae. Intergovernmental Oceanographic Commission of the UNESCO.</ref>). In order to minimise the damage to human health or living resources, such as shellfish and fish, as well as economic losses to fishermen, aquaculture and the tourist industry, efficient [[State of the Art Overview on Field Observation Techniques (Theme 9)|monitoring methods]] are required for monitoring potentially toxic algal species (identification and quantification) (Andersen et al., 2003.<ref name="A"> Andersen P., Enevoldsen H. and Anderson, D.M. (2003). Harmful algal monitoring programme and action plan design. In G.M. Hallegraeff, D.M. Anderson & A.D. Cembella (Eds.), Manual on Harmful Marine Microalgae (pp. 627-647). United Nations Educational, Scientific and Cultural Organization.</ref>).  [[Image:ALGADEC_3.jpg|thumb|right|'''Figure 2''' Schematic drawing of a sandwich hybridisation. The target organism is identified by binding of two species specific molecular probes to the ribosomal RNA (rRNA). One of the probes is immobilised on the surface of the sensorchip and the other is coupled to digoxigenin, which binds to an antibody enzyme-complex. The enzyme catalyses a redox-reaction that can be measured as an electrochemical signal.]] The identification of unicelluar algae with conventional methods like light microscopy requires a thorough taxonomic expertise and is time-consuming. It is also costly if larger numbers of samples need to be processed. In some cases toxic and non-toxic varieties (strains) belong to the same species. They are morphologically identical, and cannot be distinguished by conventional methods. Consequently, improved monitoring methods that allow rapid detection and counting of toxic algae are needed. In the past decade, a variety of molecular methods have been adapted for the detection of harmful algae. Most of these techniques focus on the genetic information in the DNA or RNA (both nucleic acids) of the organisms. However, most of these new techniques are lab-based and not suited to be carried out in the field.
 ===Methods and techniques===

@@ Regel 1: / Regel 1: @@
 {{featured}}
 ===Introduction===
-Microalgae are the major producers of [[Biomass|biomass]] and organic compounds in the aquatic environment. Among the marine microalgae there are 97 toxic species (mainly dinoflagellates) known to have the potential to form "[[Harmful_algal_blooms|Harmful Algal Blooms]]", the so called HABs (Fig. 1). [[Image:ALGADEC_1.jpg|thumb|left|'''Figure 1''' Bloom of ''Noctiluca scintillas'' in October 2002, Leigh, New Zealand.]] In recent decades, the public health and economic impacts of toxic algae species appear to have increased in frequency, intensity and geographic distribution (Zingone and Enevoldsen, 2000<ref name="Z&E">Zingone, A. & Enevoldsen, H.O. (2000). The diversity of harmful algal blooms: a challenge for science and management. Ocean & coastal management, 43, 725-748.</ref>, Daranas et al., 2001<ref name="D">Daranas A. H., Norte M. and Fernandez, J. J. (2001). Toxic marine microalgae, Toxicon, 39, 1101-1132.</ref>, Hallegraeff, 2003<ref name="H">Hallegraeff G. M. (2003). Harmful algal blooms: a global overview. In G.M. Hallegraeff, D.M. Anderson & A.D. Cembella (Eds.), Manual on Harmful Marine Microalgae (pp. 25-49). United Nations Educational, Scientific and Cultural Organization.</ref>, Moestrup, 2004.<ref name="M"> Moestrup, O. (2004). IOC Taxonomic Reference list of Toxic Algae. In O. Moestrup (Ed.), IOC taxonomic reference list of toxic algae. Intergovernmental Oceanographic Commission of the UNESCO.</ref>). In order to minimise the damage to human health or living resources, such as shellfish and fish, as well as economic losses to fisherman, aquaculture and the tourist industry, efficient [[State of the Art Overview on Field Observation Techniques (Theme 9)
+Microalgae are the major producers of [[Biomass|biomass]] and organic compounds in the aquatic environment. Among the marine microalgae there are 97 toxic species (mainly dinoflagellates) known to have the potential to form "[[Harmful_algal_blooms|Harmful Algal Blooms]]", the so called HABs (Fig. 1). [[Image:ALGADEC_1.jpg|thumb|left|'''Figure 1''' Bloom of ''Noctiluca scintillas'' in October 2002, Leigh, New Zealand.]] In recent decades, the public health and economic impacts of toxic algae species appear to have increased in frequency, intensity and geographic distribution (Zingone and Enevoldsen, 2000<ref name="Z&E">Zingone, A. & Enevoldsen, H.O. (2000). The diversity of harmful algal blooms: a challenge for science and management. Ocean & coastal management, 43, 725-748.</ref>, Daranas et al., 2001<ref name="D">Daranas A. H., Norte M. and Fernandez, J. J. (2001). Toxic marine microalgae, Toxicon, 39, 1101-1132.</ref>, Hallegraeff, 2003<ref name="H">Hallegraeff G. M. (2003). Harmful algal blooms: a global overview. In G.M. Hallegraeff, D.M. Anderson & A.D. Cembella (Eds.), Manual on Harmful Marine Microalgae (pp. 25-49). United Nations Educational, Scientific and Cultural Organization.</ref>, Moestrup, 2004.<ref name="M"> Moestrup, O. (2004). IOC Taxonomic Reference list of Toxic Algae. In O. Moestrup (Ed.), IOC taxonomic reference list of toxic algae. Intergovernmental Oceanographic Commission of the UNESCO.</ref>). In order to minimise the damage to human health or living resources, such as shellfish and fish, as well as economic losses to fisherman, aquaculture and the tourist industry, efficient [[State of the Art Overview on Field Observation Techniques (Theme 9)|monitoring methods]] are required for monitoring potentially toxic algal species (identification and quantification) (Andersen et al., 2003.<ref name="A"> Andersen P., Enevoldsen H. and Anderson, D.M. (2003). Harmful algal monitoring programme and action plan design. In G.M. Hallegraeff, D.M. Anderson & A.D. Cembella (Eds.), Manual on Harmful Marine Microalgae (pp. 627-647). United Nations Educational, Scientific and Cultural Organization.</ref>).  [[Image:ALGADEC_3.jpg|thumb|right|'''Figure 2''' Schematic drawing of a sandwich hybridisation. The target organism is identified by binding of two species specific molecular probes to the ribosomal RNA (rRNA). One of the probes is immobilised on the surface of the sensorchip and the other is coupled to digoxigenin, which binds to an antibody enzyme-complex. The enzyme catalyses a redox-reaction that can be measured as an electrochemical signal.]] The identification of unicelluar algae with conventional methods like light microscopy requires a thorough taxonomic expertise and is time-consuming. It is also costly if larger numbers of samples need to be processed. In some cases toxic and non-toxic varieties (strains) belong to the same species. They are morphologically identical, and cannot be distinguished by conventional methods. Consequently, improved monitoring methods that allow rapid detection and counting of toxic algae are needed. In the past decade, a variety of molecular methods have been adapted for the detection of harmful algae. Most of these techniques focus on the genetic information in the DNA or RNA (both nucleic acids) of the organisms. However, most of these new techniques are lab-based and not suited to be carried out in the field.
 |monitoring methods]] are required for monitoring potentially toxic algal species (identification and quantification) (Andersen et al., 2003.<ref name="A"> Andersen P., Enevoldsen H. and Anderson, D.M. (2003). Harmful algal monitoring programme and action plan design. In G.M. Hallegraeff, D.M. Anderson & A.D. Cembella (Eds.), Manual on Harmful Marine Microalgae (pp. 627-647). United Nations Educational, Scientific and Cultural Organization.</ref>).  [[Image:ALGADEC_3.jpg|thumb|right|'''Figure 2''' Schematic drawing of a sandwich hybridisation. The target organism is identified by binding of two species specific molecular probes to the ribosomal RNA (rRNA). One of the probes is immobilised on the surface of the sensorchip and the other is coupled to digoxigenin, which binds to an antibody enzyme-complex. The enzyme catalyses a redox-reaction that can be measured as an electrochemical signal.]] The identification of unicelluar algae with conventional methods like light microscopy requires a thorough taxonomic expertise and is time-consuming. It is also costly if larger numbers of samples need to be processed. In some cases toxic and non-toxic varieties (strains) belong to the same species. They are morphologically identical, and cannot be distinguished by conventional methods. Consequently, improved monitoring methods that allow rapid detection and counting of toxic algae are needed. In the past decade, a variety of molecular methods have been adapted for the detection of harmful algae. Most of these techniques focus on the genetic information in the DNA or RNA (both nucleic acids) of the organisms. However, most of these new techniques are lab-based and not suited to be carried out in the field.
 ===Methods and techniques===