| Abstract | Harmful cyanobacterial blooms have been increasing in freshwater ecosystems in recent decades, mainly because of eutrophication and climate change. Of the 150 known cyanobacteria genera, more than 40 species produce toxins, which are natural compounds showing different chemical and toxicological characteristics. Cyanobacterial toxins are responsible for both acute and chronic poisoning in animals and humans. Among the main classes of cyanotoxins, microcystins are among the most frequently found in the environment. Fast and sensitive methods to identify unequivocally Microcystis aeruginosa and Planktothrix agardhii are very useful to discriminate these species with respect to the other non-toxic cyanobacteria. For this purpose, we designed, developed and validated some oligonucleotide probes (GNPlankS02, PkAgD03, MicAerD03) for FISH (Fluorescence In-Situ Hybridization) analysis to detect these species in freshwater samples. The FISH probes were designed using the ARB software with the Silva database in the framework of the MicroCoKit project. We tested different fixative methods to minimise the natural autofluorescence from chlorophyll-a to visualize Microcystis aeruginosa and Planktothrix agardii under a laser confocal microscope. Firstly, the FISH probes designed have been tested on pure cultures of M. aeruginosa and P. agardhii species, and then the probes were successfully applied to natural samples collected from surface waters. |
| Authors | Barra Caracciolo A, Dejana L, Grenni P, Martin M, Mengs G, Saccà ML, Congestri R, Viaggiu E, Medlin L |
| Text | 398651 2018 cyanobacterial blooms FISH CARD FISH Identification of cyanobacteria in surface water by fluorescence in situ hybridization techniques Barra Caracciolo A, Dejana L, Grenni P, Martin M, Mengs G, Sacca ML, Congestri R, Viaggiu E, Medlin L Barra Caracciolo A, Dejana L, Grenni P, IRSA CNR Martin M Facultad de Veterinaria, Complutense University, Madrid, Spain Mengs G Natural Biotec S.L., Parque Cientifico de Madrid, Madrid, Spain Sacca ML Agriculture and Environment Research Center, Consiglio per la Ricerca in Agricoltura e l Analisi dell Economia Agraria, Bologna, Italy Congestri R, Viaggiu E Department of Chemical Sciences and Technologies, University of Rome Tor Vergata, Rome, Italy Medlin L Marine Biological Association of the UK, The Citadel, Plymouth, UK First Scientific Symposium Health and Climate Change Symposium, ABSTRACT BOOK W. Ricciardi, S. Marcheggiani, C. Puccinelli, M. Carere, T. Sofia, F. Giuliano, E. Dogliotti and L. Mancini Published version https //healthclimate2018.iss.it/ p=852 18 C5 First Scientific Symposium Health and Climate Change Symposium Roma 3 5/12/2018 Internazionale Contributo Harmful cyanobacterial blooms have been increasing in freshwater ecosystems in recent decades, mainly because of eutrophication and climate change. Of the 150 known cyanobacteria genera, more than 40 species produce toxins, which are natural compounds showing different chemical and toxicological characteristics. Cyanobacterial toxins are responsible for both acute and chronic poisoning in animals and humans. Among the main classes of cyanotoxins, microcystins are among the most frequently found in the environment. Fast and sensitive methods to identify unequivocally Microcystis aeruginosa and Planktothrix agardhii are very useful to discriminate these species with respect to the other non toxic cyanobacteria. For this purpose, we designed, developed and validated some oligonucleotide probes GNPlankS02, PkAgD03, MicAerD03 for FISH Fluorescence In Situ Hybridization analysis to detect these species in freshwater samples. The FISH probes were designed using the ARB software with the Silva database in the framework of the MicroCoKit project. We tested different fixative methods to minimise the natural autofluorescence from chlorophyll a to visualize Microcystis aeruginosa and Planktothrix agardii under a laser confocal microscope. Firstly, the FISH probes designed have been tested on pure cultures of M. aeruginosa and P. agardhii species, and then the probes were successfully applied to natural samples collected from surface waters. Abstract Book AbstractBook.pdf Abstract in atti di convegno Istituto superiore di sanita. 0393 5620 ISTISAN congressi ISTISAN congressi ISTISAN congr. Istituto superiore di sanita congressi Istituto superiore di sanita. 0393 5620 ISTISAN congressi ISTISAN congressi ISTISAN congr. Istituto superiore di sanita congressi anna.barracaracciolo BARRA CARACCIOLO ANNA paola.grenni GRENNI PAOLA TA.P04.005.011 Vulnerabilita degli ecosistemi delle acque sotterranee e attenuazione naturale degli inquinanti nel suolo e nel sottosuolo |
