@article{fdi:010072970,
  title = {{R}apid characterization of microcystin-producing cyanobacteria in freshwaterlakes by {TSA}-{FISH} ({T}yramid {S}ignal {A}mplification-{F}luorescent in {S}itu {H}ybridization)},
  author = {{B}rient, {L}. and {B}en {G}amra, {N}. and {P}eriot, {M}. and {R}oumagnac, {M}. and {Z}eller, {P}. and {B}ormans, {M}. and {M}{\'e}jaen, {A}. and {P}loux, {O}. and {B}iegala, {I}sabelle},
  editor = {},
  language = {{ENG}},
  abstract = {{M}icrocystin ({MC}) is a common and widespread toxin which represents a health hazard to humans and animals. {MC} toxin concentrations are monitored by various direct or proxy techniques ({HPLC}, {LC}-{MS}/{MS}, {ELISA}, {PPIA}), however, these techniques do not discriminate producing species from non-producing ones. {I}n order to simultaneously provide the identity and activity of cyanotoxin producing species in freshwater lakes, we applied simple, and fully detailed, whole cell fluorescent in situ hybridization enhanced by tyramid signal amplification ({TSA}-{FISH}). {DNA} oligonucleotide probes {MICR}3 and {MCYA} were targeting 16{S} r{RNA} and mcy{A}-m{RNA}, respectively. {T}he mcy{A} gene is coding for the {MC} synthetase enzyme involved in {MC} synthesis. {C}ontrols were acquired with the general eubacterial 16{S} r{RNA} probe {EUB}338, for {TSA}-{FISH} assay, and standard {HPLC} and {LC}-{MS}/{MS} as standard methods for the measurements of {MC} concentration. {R}esults obtained from monoclonal strains and natural samples demonstrated a specific identification of {M}icrocystis species and were able to discriminate {MC} producing from non-producing ones. {I}n addition, the {MCYA} probe allowed the specific detection of {MC}-synthetase m{RNA} within {P}lanktothrix isothrix ({O}scillatoriale) filaments. {T}wo kinds of mcy{A}-m{RNA} labeling were observed in these cells, spots like and plasmid like, which illustrates the well-known plasticity of microbial genome to adapt to environmental stresses. {W}e demonstrated that a simple {TSA}-{FISH} assay allows acquiring rapidly dual information of the presence and abundance of potentially toxic species, while identifying species actively producing {MC}-synthetase m{RNA}, a proxy of {MC} toxin. {T}his technique has the potential to be developed into an effective environmental monitoring tool. {I}n addition, detail visualization of cellular m{RNA}s is powerful for the acquisition of ecological and biomolecular studies of toxic cyanobacteria.},
  keywords = {{BACTERIE} ; {EAU} {DOUCE} ; {IDENTIFICATION} ; {TOXICITE} ; {FLUORESCENCE} ; {METHODE} {BIOLOGIQUE}},
  booktitle = {},
  journal = {{F}rontiers in {E}nvironmental {S}cience},
  volume = {5},
  numero = {},
  pages = {art. no 43 [7  ]},
  ISSN = {2296-665{X}},
  year = {2017},
  DOI = {10.3389/fenvs.2017.00043},
  URL = {https://www.documentation.ird.fr/hor/fdi:010072970},
}