First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium &amp;lt;i&amp;gt;Aphanizomenon ovalisporum&amp;lt;/i&amp;gt; in Water

Campo, Elena; Lezcano, María Ángeles; Agha, Ramsy; Cirés, Samuel; Quesada, Antonio; El-Shehawy, Rehab

doi:10.4236/aim.2013.35058

Cited by 13 publications

(7 citation statements)

References 45 publications

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“…A TaqMan-based qPCR assay for quantifying A. ovalisporum CYN production was developed by Campo et al [ 90 ]. This assay was able to discriminate CYN-producing A. ovalisporum strains from other Nostocales, such as C. raciborskii and A. bergii .…”

Section: Qpcr For Cyanotoxinsmentioning

confidence: 99%

Is qPCR a Reliable Indicator of Cyanotoxin Risk in Freshwater?

Pacheco

Guedes

Azevedo

2016

Toxins

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The wide distribution of cyanobacteria in aquatic environments leads to the risk of water contamination by cyanotoxins, which generate environmental and public health issues. Measurements of cell densities or pigment contents allow both the early detection of cellular growth and bloom monitoring, but these methods are not sufficiently accurate to predict actual cyanobacterial risk. To quantify cyanotoxins, analytical methods are considered the gold standards, but they are laborious, expensive, time-consuming and available in a limited number of laboratories. In cyanobacterial species with toxic potential, cyanotoxin production is restricted to some strains, and blooms can contain varying proportions of both toxic and non-toxic cells, which are morphologically indistinguishable. The sequencing of cyanobacterial genomes led to the description of gene clusters responsible for cyanotoxin production, which paved the way for the use of these genes as targets for PCR and then quantitative PCR (qPCR). Thus, the quantification of cyanotoxin genes appeared as a new method for estimating the potential toxicity of blooms. This raises a question concerning whether qPCR-based methods would be a reliable indicator of toxin concentration in the environment. Here, we review studies that report the parallel detection of microcystin genes and microcystin concentrations in natural populations and also a smaller number of studies dedicated to cylindrospermopsin and saxitoxin. We discuss the possible issues associated with the contradictory findings reported to date, present methodological limitations and consider the use of qPCR as an indicator of cyanotoxin risk.

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Section: Qpcr For Cyanotoxinsmentioning

confidence: 99%

Is qPCR a Reliable Indicator of Cyanotoxin Risk in Freshwater?

Pacheco

Guedes

Azevedo

2016

Toxins

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“…For our study, we have chosen previously published assays mcyE-Ana, mcyE-Mic, mcyE-Pla, cyrJ and sxtA, targeting microcystin-producers from genera Dolichospermum (ex Anabaena ), Microcystis and Planktothrix , cylindrospermopsin-producers and saxitoxin-producers, respectively ([ 6 , 15 , 16 , 17 , 18 ]; Table 1 ). First, we evaluated the selected qPCR assays in terms of their specificity, sensitivity and robustness.…”

Section: Resultsmentioning

confidence: 99%

“…We focused on water bodies in different regions of Slovenia (central Europe) and on three groups of cyanotoxins: microcystins, cylindrospermopsins and saxitoxins. We employed five previously published qPCR assays for detection of the microcystin- [ 15 , 16 , 17 ], cylindrospermopsin- [ 18 ] and saxitoxin-producing cyanobacteria [ 6 ]. Although the cyanotoxin potential is not necessarily linked to cyanotoxin concentration, we evaluated the correlation between the number of gene copies, microscopically determined cell number of potentially toxic species and cyanotoxin concentration.…”

Section: Introductionmentioning

confidence: 99%

Potentially Toxic Planktic and Benthic Cyanobacteria in Slovenian Freshwater Bodies: Detection by Quantitative PCR

Zupančič

Kogovšek

Šter

et al. 2021

Toxins

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Due to increased frequency of cyanobacterial blooms and emerging evidence of cyanotoxicity in biofilm, reliable methods for early cyanotoxin threat detection are of major importance for protection of human, animal and environmental health. To complement the current methods of risk assessment, this study aimed to evaluate selected qPCR assays for detection of potentially toxic cyanobacteria in environmental samples. In the course of one year, 25 plankton and 23 biofilm samples were collected from 15 water bodies in Slovenia. Three different analyses were performed and compared to each other; qPCR targeting mcyE, cyrJ and sxtA genes involved in cyanotoxin production, LC-MS/MS quantifying microcystin, cylindrospermopsin and saxitoxin concentration, and microscopic analyses identifying potentially toxic cyanobacterial taxa. qPCR analyses detected potentially toxic Microcystis in 10 lake plankton samples, and potentially toxic Planktothrix cells in 12 lake plankton and one lake biofilm sample. A positive correlation was observed between numbers of mcyE gene copies and microcystin concentrations. Potential cylindrospermopsin- and saxitoxin-producers were detected in three and seven lake biofilm samples, respectively. The study demonstrated a potential for cyanotoxin production that was left undetected by traditional methods in both plankton and biofilm samples. Thus, the qPCR method could be useful in regular monitoring of water bodies to improve risk assessment and enable timely measures.

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“…quantitative PCR-qPCR) have been applied since the 1980 s and 1990 s in the analysis of microbial DNA, including ''phytoplankton'' (for a review, see Wilmotte et al, 2017). These approaches are tuned to target common regions of the whole genomic DNA extracted from water samples or other substrata, providing information on the existence of specific taxonomic and toxins encoding genes (Campo et al, 2013;Capelli et al, 2018), and the taxonomic composition of the algal community without the need to isolate and cultivate individual strains. In this latter group of methods, probably one of the most used in phytoplankton ecology is the denaturing gradient gel electrophoresis (DGGE; (Strathdee & Free, 2013).…”

Section: Culture Independent Approaches-traditional Methodsmentioning

confidence: 99%

Freshwater phytoplankton diversity: models, drivers and implications for ecosystem properties

et al. 2020

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Our understanding on phytoplankton diversity has largely been progressing since the publication of Hutchinson on the paradox of the plankton. In this paper, we summarise some major steps in phytoplankton ecology in the context of mechanisms underlying phytoplankton diversity. Here, we provide a framework for phytoplankton community assembly and an overview of measures on taxonomic and functional diversity. We show how ecological theories on species competition together with modelling approaches and laboratory experiments helped understand species coexistence and maintenance of diversity in phytoplankton. The nonequilibrium nature of phytoplankton and the role of disturbances in shaping diversity are also discussed. Furthermore, we discuss the role of water body size, productivity of habitats and temperature on phytoplankton species richness, and how diversity may affect the functioning of lake ecosystems. At last, we give an insight into molecular tools that have emerged in the last decades and argue how it has broadened our perspective on microbial diversity. Besides historical backgrounds, some critical comments have also been made.

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First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium <i>Aphanizomenon ovalisporum</i> in Water

Cited by 13 publications

References 45 publications

Is qPCR a Reliable Indicator of Cyanotoxin Risk in Freshwater?

Is qPCR a Reliable Indicator of Cyanotoxin Risk in Freshwater?

Potentially Toxic Planktic and Benthic Cyanobacteria in Slovenian Freshwater Bodies: Detection by Quantitative PCR

Freshwater phytoplankton diversity: models, drivers and implications for ecosystem properties

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