Brevetoxin metabolism and elimination in the Eastern oyster (Crassostrea virginica) after controlled exposures to Karenia brevis

Plakas, Steven M.; Wang, Zhihong; Said, Kathleen R. El; Jester, Edward L.E.; Granade, Hudson R.; Flewelling, Leanne J.; Scott, Paula S.; Dickey, Robert

doi:10.1016/j.toxicon.2004.07.027

Cited by 69 publications

(67 citation statements)

References 17 publications

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“…These organisms have gained recognition as fish-killing phytoplankton, causing massive mortalities of fish and resulting in millions of dollars in damage to the aquaculture industry (Black et al 1991;Horner 1999;Yang et al 1995). In addition, brevetoxin-like compounds produced by Raphidophytes (Haque and Onoue 2002;Khan et al 1996aKhan et al , 1996bKhan et al , 1997) pose a threat to higher trophic levels (including wildlife and humans) since they can potentially be concentrated during food web transfer (Ishida et al 2004;Plakas et al 2004;Stommel and Watters 2004;Woofter et al 2005). Conventional microscopic methods for identifying and estimating the abundance of Raphidophyte species in complex environmental samples are time-consuming and often lack the sensitivity required for background level detection.…”

mentioning

confidence: 99%

Improved quantitative real‐time PCR assays for enumeration of harmful algal species in field samples using an exogenous DNA reference standard

Coyne

Handy

Demir

et al. 2005

Limnology & Ocean Methods

137

152

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Quantitative real-time PCR (QPCR) is a powerful and sensitive method for quantitative detection of microorganisms. Application of this methodology for enumeration of harmful algal bloom (HAB) species has the potential to revolutionize our approach to HAB research, making it possible to identify correlations between cell abundances and factors that regulate bloom dynamics. Its application to ecological studies, however, has produced mixed results. QPCR assays typically rely on the generation of standard curves from plasmids or laboratory cultures that may be unrealistic when compared to amplification of DNA extracted from field samples. In addition, existing methods often fail to incorporate controls to assess variability in extraction and amplification efficiencies, or include controls that are sequence-specific and preclude the investigation of multiple species. Here, we describe the development and rigorous analysis of QPCR assays for two HAB species, Chattonella subsalsa and Heterosigma akashiwo, in which we introduce a known concentration of exogenous DNA plasmid into the extraction buffer as a reference standard. Since the target DNA is extracted in the presence of the reference standard, inherent variability in extraction and amplification efficiencies affect both target and standard equally. Furthermore, the reference standard is applicable to QPCR analysis of any microbial species. Using environmental bloom samples as calibrators, we evaluated the accuracy of the comparative Ct method for enumeration of target species in several field samples. Our investigation demonstrates that the comparative Ct method with an exogenous DNA reference standard provides both accurate and reproducible quantification of HAB species in environmental samples.

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mentioning

confidence: 99%

Improved quantitative real‐time PCR assays for enumeration of harmful algal species in field samples using an exogenous DNA reference standard

Coyne

Handy

Demir

et al. 2005

Limnology & Ocean Methods

137

152

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“…(S-desoxy-Btx-B2 and Btx-B2) antibody to other Btx was reported by several groups (Dechraoui et al, 2007;Maucher et al, 2007;Plakas et al, 2004), thereby raising some doubt as to the specificity of this antibody. Due to recruitment of a magneto-controlled immunosensing platform a lOD of 1 pg Btx-2/ml extract with a range of 1 pg/ml to 10 ng/ml could be reached (tang et al, 2012).…”

Section: Neurotoxic Shellfish Poisoning (Nsp)mentioning

confidence: 87%

“…Functional, biochemical and biomolecular assays lC-MS/MS methodologies have been developed, evaluated and have been used for detection of Btx since the mid 1990s (Dickey et al, 1999Fire et al, 2008;Hua and Cole, 2000;Hua et al, 1995;Ishida et al, 2004a,b,c;Nozawa et al, 2003;Pierce et al, 2006;Plakas et al, 2002Plakas et al, , 2008Plakas et al, , 2004Wang and Cole, 2009;Wang et al, 2004;McNabb et al, 2012). the reported LOQ are 2, 0.2, 0.4, and 0.4 µg/kg shellfish meat for BTX-2, Btx-3, Btx-B5 and Btx-B1, respectively.…”

Section: )mentioning

confidence: 99%

A roadmap for hazard monitoring and risk assessment of marine biotoxins on the basis of chemical and biological test systems

Daneshian

2013

ALTEX

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Summary Aquatic food accounts for over 40% of global animal food products, and the potential contamination with toxins of algal origin -marine biotoxins -poses a health threat for consumers. The gold standards to assess toxins in aquatic food have traditionally been in vivo methodssingle marine biotoxins or combinations of marine biotoxins and intoxication symptomatology as well as monitoring the "success" of controls implemented during fishery and aquaculture production, processing, and retailing. For this reason, a workshop was organized in ermatingen, Switzerland by the Center for Alternatives to Animal testing -europe (CAAteurope) within the framework of the transatlantic think tank for toxicology (t 4 ).In contrast to other food products where hygiene and potential contamination with microbes or mold toxins is the prime issue, the emphasis in aquatic products (shellfish and finfish) is, beyond viral and bacterial contaminations, primarily placed on potential contamination with marine biotoxins owing to the numerous and recurring human intoxications.The gold standards to assess toxins in aquatic food have traditionally been in vivo methods, i.e., the mouse and the rat bioassays. Besides the ethical issues of in vivo bioassays there are specific difficulties, e.g., different exposure route (i.p. versus oral), low inter-species comparability, high intra-species variability, and questionable extrapolation of quantitative risk to humans, thus highlighting the need for the development of more reliable detection and quantification methods. Based on this reasoning, the european Food Safety Authority (eFSA) advocates the use of an analytical method, i.e., LC-MS (Liquid Chromatography coupled Mass Spectrometry), as a substitute for in vivo bioassays for almost all classes of marine toxins. However, although lC-MS is a very sensitive method that has the advantages of being able to detect multiple toxins in a single analysis and being good for confirming the identity of toxins, the quality of quantitative analysis is dependent on the availability of calibration standards. While many new toxin structural analogues have been detected and identified using LC-MS, this technique -as with most other methods -is not good at detecting new toxin types. When new toxin analogues are detected but accurate standards are not yet available, only an approximate quantitation is possible using estimated response factors.For the purpose of risk assessment of food, however, it is imperative to focus on the possible adverse effects, independent of whether they stem from one toxin or a combination of toxins. As toxicity is defined by functional and structural changes of biological systems, the development of a human-relevant in vitro system for appropriate risk assessment of marine biotoxins in seafood stands to reason. Moreover, poor correlation of human intoxications, of acute symptomatology and long-term adverse effects, and of predictive in vitro, in vivo, and analytical detection methodologies of marine biotoxins stems not least from a...

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“…In LC-MS, brevetoxin congeners are chromatographically separated and individually quantified by a mass-tocharge ratio (m/z) relative to standard reference material, and this method was utilized as a confirmatory method for ELISA and RBA data. Selected samples were initially analyzed for PbTx-3 following LC-MS methods described by Plakas et al (2004), using a ThermoFinnigan AqA HPLC-MS equipped with an AqA single quad system, with a limit of quantification of 300 ng g -1…”

Section: Methodsmentioning

confidence: 99%

Prevalence of brevetoxins in prey fish of bottlenose dolphins in Sarasota Bay, Florida

Fire¹,

Flewelling²,

Naar³

et al. 2008

Mar. Ecol. Prog. Ser.

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Blooms of the brevetoxin-producing dinoflagellate Karenia brevis have been linked to high mortality of bottlenose dolphins Tursiops truncatus on Florida's Gulf of Mexico coast. A clear understanding of trophic transfer of brevetoxin from its algal source up the food web to top predators is needed to assess exposure of affected dolphin populations. Prey fish constitute a means of accumulating and transferring brevetoxins and are potential vectors of brevetoxin to dolphins frequently exposed to K. brevis blooms. Here we report results of brevetoxin analyses of the primary fish species consumed by long-term resident bottlenose dolphins inhabiting Sarasota Bay, Florida. Fish collected during K. brevis blooms in 2003 to 2006 were analyzed by competitive enzyme-linked immunosorbent assay (ELISA) and had brevetoxin concentrations ranging from 4 to 10 844 ng PbTx-3 eq g -1 tissue. Receptor binding assay (RBA) and liquid chromatography-mass spectrometry (LC-MS) analysis confirmed toxicity and the presence of parent brevetoxins and known metabolites. Fish collected in the absence of K. brevis blooms tested positive for brevetoxin by ELISA and RBA, with concentrations up to 1500 ng PbTx-3 eq g -1 tissue. These findings implicate prey fish exposed to K. brevis blooms as brevetoxin vectors for their dolphin predators and provide a critical analysis of persistent brevetoxin loads in the food web of dolphins repeatedly exposed to Florida red tides.

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Brevetoxin metabolism and elimination in the Eastern oyster (Crassostrea virginica) after controlled exposures to Karenia brevis

Cited by 69 publications

References 17 publications

Improved quantitative real‐time PCR assays for enumeration of harmful algal species in field samples using an exogenous DNA reference standard

Improved quantitative real‐time PCR assays for enumeration of harmful algal species in field samples using an exogenous DNA reference standard

A roadmap for hazard monitoring and risk assessment of marine biotoxins on the basis of chemical and biological test systems

Prevalence of brevetoxins in prey fish of bottlenose dolphins in Sarasota Bay, Florida

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