qPCR Assays for the Detection and Quantification of Multiple Paralytic Shellfish Toxin-Producing Species of Alexandrium

Ruvindy, Rendy; Bolch, Christopher J. S.; MacKenzie, Lincoln; Smith, Kirsty F.; Murray, Shauna A.

doi:10.3389/fmicb.2018.03153

Cited by 37 publications

(27 citation statements)

References 56 publications

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“…qPCR analysis was carried out using assays specific to A. catenella , A. pacificum , and A. australiense [ 31 ]. Species-specific primers were used to identify the species present in the sample ( Table 2 ).…”

Section: Methodsmentioning

confidence: 99%

First Detection of Paralytic Shellfish Toxins from Alexandrium pacificum above the Regulatory Limit in Blue Mussels (Mytilus galloprovincialis) in New South Wales, Australia

et al. 2020

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In 2016, 2017 and 2018, elevated levels of the species Alexandrium pacificum were detected within a blue mussel (Mytilus galloprovincialis) aquaculture area at Twofold Bay on the south coast of New South Wales, Australia. In 2016, the bloom persisted for at least eight weeks and maximum cell concentrations of 89,000 cells L−1 of A. pacificum were reported. The identity of A. pacificum was confirmed using molecular genetic tools (qPCR and amplicon sequencing) and complemented by light and scanning electron microscopy of cultured strains. Maximum reported concentrations of paralytic shellfish toxins (PSTs) in mussel tissue was 7.2 mg/kg PST STX equivalent. Elevated cell concentrations of A. pacificum were reported along the adjacent coastal shelf areas, and positive PST results were reported from nearby oyster producing estuaries during 2016. This is the first record of PSTs above the regulatory limit (0.8 mg/kg) in commercial aquaculture in New South Wales since the establishment of routine biotoxin monitoring in 2005. The intensity and duration of the 2016 A. pacificum bloom were unusual given the relatively low abundances of A. pacificum in estuarine and coastal waters of the region found in the prior 10 years.

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Section: Methodsmentioning

confidence: 99%

First Detection of Paralytic Shellfish Toxins from Alexandrium pacificum above the Regulatory Limit in Blue Mussels (Mytilus galloprovincialis) in New South Wales, Australia

et al. 2020

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“…Several qPCR assays for the detection of Alexandrium spp. have been developed based on rRNA genes (e.g., References [216,217,218,219,220,221]), with specificities and sensitivities down to one cell per litre [221]. However, these assays do not provide any indication regarding the potential toxicity of a bloom as they only detect the presence of the Alexandrium cells.…”

Section: Dinoflagellate Toxinsmentioning

confidence: 99%

“…Furthermore, studies have shown that rRNA gene copy numbers are highly variable [216,217,222], and this may lead to over- or underestimation of cell densities [221,223]. Characterization of sxt genes has made the development of PST-producing strain specific qPCR assays [214,215,224,225], and studies have shown that the genomic copy numbers of sxt genes vary less compared to rRNA genes, allowing for more accurate cell density estimates [197,223,224].…”

Section: Dinoflagellate Toxinsmentioning

confidence: 99%

The Genetic Basis of Toxin Biosynthesis in Dinoflagellates

et al. 2019

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In marine ecosystems, dinoflagellates can become highly abundant and even dominant at times, despite their comparatively slow growth rates. One factor that may play a role in their ecological success is the production of complex secondary metabolite compounds that can have anti-predator, allelopathic, or other toxic effects on marine organisms, and also cause seafood poisoning in humans. Our knowledge about the genes involved in toxin biosynthesis in dinoflagellates is currently limited due to the complex genomic features of these organisms. Most recently, the sequencing of dinoflagellate transcriptomes has provided us with valuable insights into the biosynthesis of polyketide and alkaloid-based toxin molecules in dinoflagellate species. This review synthesizes the recent progress that has been made in understanding the evolution, biosynthetic pathways, and gene regulation in dinoflagellates with the aid of transcriptomic and other molecular genetic tools, and provides a pathway for future studies of dinoflagellates in this exciting omics era.

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“…By including standard curves of target DNA, the sensitive, qualitative detection is complemented with quantitative estimations of the target organism. Currently, the detection and quantification of toxic microalgae via qPCR has become a standard procedure in many studies and monitoring programs [28][29][30].…”

Section: Introductionmentioning

confidence: 99%

Distribution and abundance of azaspiracid-producing dinophyte species and their toxins in North Atlantic and North Sea waters in summer 2018

et al. 2020

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Representatives of the marine dinophyte family Amphidomataceae produce lipophilic phycotoxins called azaspiracids (AZA) which may cause azaspiracid shellfish poisoning (AZP) in humans after consumption of contaminated seafood. Three of the four known toxigenic species are observed frequently in the eastern North Atlantic. In 2018, a research survey was performed to strengthen knowledge on the distribution and abundance of toxigenic Amphidomataceae and their respective toxins in Irish coastal waters and in the North Sea. Species-specific quantification of the three toxigenic species (Azadinium spinosum, Azadinium poporum and Amphidoma languida) was based on recently developed qPCR assays, whose performance was successfully validated and tested with specificity tests and spike experiments. The multi-method approach of on-board live microscopy, qPCR assays and chemical AZA-analysis revealed the presence of Amphidomataceae in the North Atlantic including the three targeted toxigenic species and their respective AZA analogues (AZA-1,-2,-33,-38,-39). Azadinium spinosum was detected at the majority of Irish stations with a peak density of 8.3 x 10 4 cells L-1 and AZA (AZA-1,-2,-33) abundances up to 1,274 pg L-1. Amphidoma languida was also present at most Irish stations but appeared in highest abundance in a bloom at a central North Sea station with a density of 1.2 x 10 5 cells L-1 and an AZA (AZA-38,-39) abundances of 618 pg L-1. Azadinium poporum was detected sporadically at the Irish south coast and North Sea and was rather low in abundance during this study. The results confirmed the wide distribution and frequent occurrence of the target species in the North Atlantic area and revealed, for the first time, bloom abundances of toxigenic Amphidomataceae in this area. This emphasizes the importance of future studies and monitoring of amphidomatacean species and their respective AZA analogues in the North Atlantic.

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qPCR Assays for the Detection and Quantification of Multiple Paralytic Shellfish Toxin-Producing Species of Alexandrium

Cited by 37 publications

References 56 publications

First Detection of Paralytic Shellfish Toxins from Alexandrium pacificum above the Regulatory Limit in Blue Mussels (Mytilus galloprovincialis) in New South Wales, Australia

First Detection of Paralytic Shellfish Toxins from Alexandrium pacificum above the Regulatory Limit in Blue Mussels (Mytilus galloprovincialis) in New South Wales, Australia

The Genetic Basis of Toxin Biosynthesis in Dinoflagellates

Distribution and abundance of azaspiracid-producing dinophyte species and their toxins in North Atlantic and North Sea waters in summer 2018

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