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Int. J. Environ. Res. Public Health 2015, 12(5), 4921-4941; doi:10.3390/ijerph120504921

Molecular Detection of a Potentially Toxic Diatom Species

1
Laboratory of Genetics, School of Biosciences and Veterinary Medicine, University of Camerino, Camerino 62032, Italy
2
Environmental, Quality and Fishfarm Unit, Environment & Primary Prevention Department, Istituto Superiore di Sanità, Viale Regina Elena, Rome 299, Italy
*
Author to whom correspondence should be addressed.
Academic Editor: Paul B. Tchounwou
Received: 15 February 2015 / Revised: 24 March 2015 / Accepted: 16 April 2015 / Published: 6 May 2015
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Abstract

A few diatom species produce toxins that affect human and animal health. Among these, members of the Pseudo-nitzschia genus were the first diatoms unambiguously identified as producer of domoic acid, a neurotoxin affecting molluscan shell-fish, birds, marine mammals, and humans. Evidence exists indicating the involvement of another diatom genus, Amphora, as a potential producer of domoic acid. We present a strategy for the detection of the diatom species Amphora coffeaeformis based on the development of species-specific oligonucleotide probes and their application in microarray hybridization experiments. This approach is based on the use of two marker genes highly conserved in all diatoms, but endowed with sufficient genetic divergence to discriminate diatoms at the species level. A region of approximately 450 bp of these previously unexplored marker genes, coding for elongation factor 1-a (eEF1-a) and silicic acid transporter (SIT), was used to design oligonucleotide probes that were tested for specificity in combination with the corresponding fluorescently labeled DNA targets. The results presented in this work suggest a possible use of this DNA chip technology for the selective detection of A. coffeaeformis in environmental settings where the presence of this potential toxin producer may represent a threat to human and animal health. In addition, the same basic approach can be adapted to a wider range of diatoms for the simultaneous detection of microorganisms used as biomarkers of different water quality levels. View Full-Text
Keywords: diatoms; domoic acid; oligonucleotide probes; microarrays diatoms; domoic acid; oligonucleotide probes; microarrays
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Dhar, B.C.; Cimarelli, L.; Singh, K.S.; Brandi, L.; Brandi, A.; Puccinelli, C.; Marcheggiani, S.; Spurio, R. Molecular Detection of a Potentially Toxic Diatom Species. Int. J. Environ. Res. Public Health 2015, 12, 4921-4941.

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