References of "Boutte, Christophe"
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See detailSeasonal cyanobacterial dynamics in a mesoeutrophic reservoir: microscopic counts and DGGE (Denaturing Gradient Gel Electrophoresis)
Willame, Raphael; Boutte, Christophe; Grubisic, Stana ULg et al

in Algological Studies (2009), 129

The seasonal planktic cyanobacterial dynamics was assessed during the year 2000 by microscopic and DGGE techniques, on the basis of 22 samples collected from the Haute-Sûre reservoir (Grand-Duchy of ... [more ▼]

The seasonal planktic cyanobacterial dynamics was assessed during the year 2000 by microscopic and DGGE techniques, on the basis of 22 samples collected from the Haute-Sûre reservoir (Grand-Duchy of Luxembourg). Microscopic investigations were carried out according to the standard Utermöhl procedure while 16S rRNA gene fragments obtained by semi-nested PCR were subsequently separated by DGGE. Sequencing of selected excised bands was performed to genotypically define the cyanobacterial assemblages. The dynamics of cyanobacterial communities obtained by both approaches were compared. Several discordances were pointed out. The counting procedure failed to detect cyanobacteria with small dimensions or in very low abundances, whereas DGGE had a lower detection limit when cyanobacteria were scarce (e.g. in spring) and performed better for the study of picosized forms. Generally, only the dominant cyanobacteria were revealed by these two methods. Actually, both techniques appeared to be complementary rather than equivalent. This study underlines the necessity to use multidisciplinary approaches to obtain a more complete view of the microbial diversity and of the community structure [less ▲]

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See detailA new approach to analyze genotypes of colony-forming cyanobacteria from environmental samples.
Lara, Yannick ULg; BOUTTE, Christophe; PERETYATKO, Anatoly et al

Poster (2008, August 31)

Several studies have shown the efficiency of sequences as rRNA-ITS, cpcBA, rbcLX and other housekeeping genes to study taxonomy [1, 2, 3], population, community structure of cyanobacteria, or for Multi ... [more ▼]

Several studies have shown the efficiency of sequences as rRNA-ITS, cpcBA, rbcLX and other housekeeping genes to study taxonomy [1, 2, 3], population, community structure of cyanobacteria, or for Multi Locus Sequence Analysis [4]. Recently, the genotypic analysis of single colonies and single filaments directly isolated from the environment has been carried out by other authors. It appears that different genotypes of Microcystis are present in one population in one lake. Besides, succession of toxic and non-toxic genotypes may have a critical influence on toxin concentrations during the blooms [5]. Genotypic analysis of colony-forming cyanobacteria requires enough DNA. So far, the genotypes of environmental single colonies of Microcystis were characterized on the basis of one or two PCR [6]. As the DNA content of one single colony only allows for a few PCR reactions, we have developed a new approach using Whole Genome Amplification with Phi29 polymerase to allow for the Multi Locus Sequences Typing analysis of a single colony or filament. For the first time, we were able to amplify and sequence more than one locus of the genome of a single colony of Microcystis. In addition, we have obtained the first sequences of rpoC1, rbcLX and rRNA-ITS from a single colony of the genus Woronichinia ( identified by microscopy). This approach allows to work with a small amount of DNA, and represents a concrete answer to the lack of data on non-cultivable cyanobacteria. This research is supported by the Belgian Science policy under the science for a sustainable development (SSD) and Fonds de la Recherche Scientifique-FNRS with a FRIA fellowship. References: [1] Otsuka S, et al (1999) FEMS Microbiology Letters 172 15-21 [2] Gugger M, et al (2002) Int J Syst Evol Microbiol 52 1867-1880 [3] Haverkamp T, et al (2008) Environmental Microbiology 10(1) 174-188 [4] Lodders N, et al (2005) Environmental Microbiology 7 (3) 434-442 [5] Kardinaal E, and Visser P (2005) In Harmfuf cyanobacteria, Springer Dordrecht pp 41-64 [6] Janse I, et al (2004) Appl Environm Microbiol 70 (7) 3979–3987 [less ▲]

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See detailDiversity of planktonic cyanobacteria and microcystin occurrence in Polish water bodies investigated using a polyphasic approach
Boutte, Christophe; Mankiewicz-Boczek, Joanna; Komarkova, Jarka et al

in Aquatic Microbial Ecology (2008), 51

Microscopic measurements of fresh biomass and 16S rRNA gene sequences from clone libraries and denaturing gradient gel electrophoresis (DGGE) were used to investigate cyanobacterial diversity in Polish ... [more ▼]

Microscopic measurements of fresh biomass and 16S rRNA gene sequences from clone libraries and denaturing gradient gel electrophoresis (DGGE) were used to investigate cyanobacterial diversity in Polish water bodies in 2002. In addition, measurements of microcystin (MC) concentrations were made. Thirty water samples were taken from 11 water bodies; of these samples, 18 were obtained from the Sulejow Reservoir during regular monitoring from June to October. Intraand extracellular MC concentrations in Sulejow samples were measured by high performance liquid chromatography (HPLC). The extracellular MC concentration was assessed using a protein phosphatase inhibition assay (PPIA) in additional lakes. Additionally, physicochemical parameters were measured (total nitrogen [TN], total phosphorus [TP], TN:TP ratio, chlorophyll a concentration, temperature). In Sulejow, high intracellular MC concentrations corresponded to large cyanobacterial biovolumes and to low TN:TP ratios. In the other lakes, extracellular MCs were not linked to any measured parameters. The combination of the microscopic and molecular data showed that Aphanizomenon and Microcystis were the dominant genera during the summer period in the Sulejow Reservoir. At the genetic level, there was a succession of 2 different operational taxonomic units (OTUs) belonging to the lineage Anabaena/Aphanizomenon. In the other water bodies, the most frequent populations were Aphanizomenon, Anabaena, Microcystis and Planktothrix. Small populations of Romeria, Snowella, Woronichinia, Limnothrix and Pseudanabaena were observed, and an enigmatic cluster affiliated with Prochlorothrix was genetically retrieved. Anabaena and Microcystis were presumed to be the main genera responsible for the MC production. [less ▲]

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See detailDiversité moléculaire des cyanobactéries planctonniques dans les eaux de surface belges
Lara, Yannick ULg; Lambion, Alexandre ULg; Simon, Patricia ULg et al

Poster (2008, April 02)

Les développements massifs de cyanobactéries ou ‘blooms’ sont devenus un phénomène récurrent et de plus en plus important dans les eaux douces du monde entier durant la dernière décennie. Ces ... [more ▼]

Les développements massifs de cyanobactéries ou ‘blooms’ sont devenus un phénomène récurrent et de plus en plus important dans les eaux douces du monde entier durant la dernière décennie. Ces efflorescences présentent des risques potentiels majeurs pour la santé humaine et animale et interfèrent négativement avec l'utilisation des eaux de surface par exemple, pour le captage d'eau potable, les loisirs nautiques, l'irrigation, les exploitations piscicoles. Entre 25 et 70% des blooms sont toxiques. Comme beaucoup de pays la Belgique n'a pas échappé au problème des efflorescences de cyanobactéries toxiques, mais il y a encore relativement peu de données. Durant la dernière décennie, trois projets européens et nationaux (MIDICHIP 1999-2003, B-BLOOMS 2003-2005, B-BLOOMS 2 2007-2011) se sont intéressés à la diversité des cyanobactéries dans les eaux de surfaces belges. Nous présentons ici un arbre phylogénétique élaboré à partir d’ un pool de 249 séquences partielles du gène codant pour l’ARNr 16S obtenu à partir de 31 échantillons d’eaux belges issus de ces projets. Cet arbre représente la mise à jour d’une base de données qui constitue l’inventaire des cyanobactéries d’eaux douces belges. Cette base de données permet le suivi de l’évolution de la diversité observable de ces organismes en Belgique et la surveillance de l’apparition d’espèces tropicales comme conséquence aux changements climatiques globaux. [less ▲]

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See detailCovariation between zooplankton community composition and cyanobacterial community dynamics in Lake Blaarmeersen (Belgium)
van Gremberghe, Ineke; Van Wichelen, Jeroen; Van der Gucht, Kathleen et al

in FEMS Microbiology Ecology (2008), 63

The cyanobacterial community composition in the mesotrophic Lake Blaarmeersen was determined by denaturing gradient gel electrophoresis (DGGE) of PCRamplified 16S rRNA gene fragments during two ... [more ▼]

The cyanobacterial community composition in the mesotrophic Lake Blaarmeersen was determined by denaturing gradient gel electrophoresis (DGGE) of PCRamplified 16S rRNA gene fragments during two consecutive years to assess the importance of different classes of explanatory variables (bottom-up and top-down factors, physical variables and phytoplankton) in cyanobacterial community dynamics. The most dominant cyanobacteria in Lake Blaarmeersen were Synechococcus (three genotypes), Limnothrix redekei and Anabaena/Aphanizomenon. Analyses of Similarity revealed that the cyanobacterial community in Lake Blaarmeersen differed significantly between the growing season and the winter season as well as between the epilimnion and hypolimnion during the stratified periods. Mantel tests revealed significant correlations between the DGGE data and bottom-up factors, physical variables, the phytoplankton community composition and, interestingly, the zooplankton community composition. In general, the zooplankton community composition (especially the cladoceran community) was more important in structuring the cyanobacterial community than the total zooplankton biomass. This study shows that grazing zooplankton communities can have a relatively strong impact on the cyanobacterial community dynamics and that this impact can be equally important as bottom-up processes regulated by nutrient concentrations and/or physical variables. [less ▲]

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See detailA polyphasic approach to assess the cyanobacterial diversity of summer samples from Czech reservoirs
Boutte, Christophe; Komarkova, Jarka; Grubisic, Stana ULg et al

in Algological Studies (2005), 117

We used a polyphasic approach combining data from microscopic assessment of fresh biomass and from clone libraries and DGGE fingerprints based on 16S rRNA gene sequences to investigate the cyanobacterial ... [more ▼]

We used a polyphasic approach combining data from microscopic assessment of fresh biomass and from clone libraries and DGGE fingerprints based on 16S rRNA gene sequences to investigate the cyanobacterial diversity of Czech reservoirs during the summer in 2001 and 2002. In total, 15 genera were identified using the microscopic analysis in 38 samples analysed. They were Aphanizomenon, Anabaena, Anabaenopsis, Aphanocapsa, Aphanothece, Pseudanabaena, Planktothrix, Planktolyngbya, Limnothrix, Woronichinia, Snowella, Romeria, Microcystis, Merismopedia, and Coelomoron. We recovered 113 DGGE band sequences from the same samples. In addition, 128 partial 16S rRNA sequences were obtained from two clone libraries of reservoirs Pilská and Orlík. The phylogenetic comparison with the currently available rRNA sequences in databases showed that our sequences belonged to 8 clusters: Woronichinia, Microcystis, Synechococcus, Snowella, Planktothrix, Anabaena/Aphanizomenon, Limnothrix and a plastid related to Chrysochromulina polylepis. The microscopic enumeration and the molecular results were generally congruent concerning the major populations determined in these samples (for 32 samples among 38). Anabaena/Aphanizomenon, Microcystis and Woronichinia were the major genera in the Czech reservoirs during summer, and were present in most of the samples. This study showed some discrepancies between the genera retrieved by the traditional method and the molecular analyses. Differences concerned the presence of minor populations belonging to Aphanothece, Romeria, Merismopedia, Synechococcus, Snowella and Pseudanabena. These differences could be explained by biases specific to each method (competitive amplification, difficulty to obtain sequences from DGGE bands, not precise microscopic observation of the small-sized genera). [less ▲]

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See detail(WO/2004/104211) METHOD FOR DETECTING TOXIC AND NON-TOXIC CYANOBACTERIA
SIVONEN, Kaarina; RANTALA, Anne; Rouhianen, Leo et al

Patent (2004)

This invention is related to a method for detecting toxic and non-toxic cyanobacteria. The method comprises that nucleic acid from a biological sample is brought into contact with an oligonucleotide ... [more ▼]

This invention is related to a method for detecting toxic and non-toxic cyanobacteria. The method comprises that nucleic acid from a biological sample is brought into contact with an oligonucleotide designed to be specific for particular regions of the mcyE gene, the mcyE in combination with mcyD, and with an oligonucleotide designed to be specific for 16SrDNA, and the presence or absence of toxic cyanobacteria is detected by a suitable molecular biology method. The invention is related also to oligonucleotides used in the method. [less ▲]

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See detailDevelopment of a universal microarray based on the ligation detection reaction and 16S rRNA gene polymorphism to target diversity of cyanobacteria
Castiglioni, Bianca; Rizzi, Ermann; Frosini, Andrea et al

in Applied and Environmental Microbiology (2004), 70(12), 7161-7172

The cyanobacteria are photosynthetic prokaryotes of significant ecological and biotechnological interest, since they strongly contribute to primary production and are a rich source of bioactive compounds ... [more ▼]

The cyanobacteria are photosynthetic prokaryotes of significant ecological and biotechnological interest, since they strongly contribute to primary production and are a rich source of bioactive compounds. In eutrophic fresh and brackish waters, their mass occurrences (water blooms) are often toxic and constitute a high potential risk for human health. Therefore, rapid and reliable identification of cyanobacterial species in complex environmental samples is important. Here we describe the development and validation of a microarray for the identification of cyanobacteria in aquatic environments. Our approach is based on the use of a ligation detection reaction coupled to a universal array. Probes were designed for detecting 19 cyanobacterial groups including Anabaena/Aphanizomenon, Calothrix, Cylindrospermopsis, Cylindrospermum, Gloeothece, halotolerants, Leptolyngbya, Palau Lyngbya, Microcystis, Nodularia, Nostoc, Planktothrix, Antarctic Phormidium, Prochlorococcus, Spirulina, Synechococcus, Synechocystis, Trichodesmium, and Woronichinia. These groups were identified based on an alignment of over 300 cyanobacterial 16S rRNA sequences. For validation of the microarrays, 95 samples (24 axenic strains from culture collections, 27 isolated strains, and 44 cloned fragments recovered from environmental samples) were tested. The results demonstrated a high discriminative power and sensitivity to 1 fmol of the PCR-amplified 16S rRNA gene. Accurate identification of target strains was also achieved with unbalanced mixes of PCR amplicons from different cyanobacteria and an environmental sample. Our universal array method shows great potential for rapid and reliable identification of cyanobacteria. It can be easily adapted to future development and could thus be applied both in research and environmental monitoring. [less ▲]

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