High-Performance Liquid Chromatography of Domoic Acid, a Marine Neurotoxin, with Application to Shellfish and Plankton

Quilliam, Michael A.; Sim, P. G.; Mcculloch, A. W.; McInnes, A. G.

doi:10.1080/03067318908026867

Cited by 145 publications

(52 citation statements)

References 11 publications

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“…The samples were analysed using a slightly modified HPLC-UV method according to Quilliam et al (1989), in which the same Develosil ODS-5 column as described in Expt 1 and a mobile phase of 10% acetonitrile in phosphate buffer (pH 2.5) were used. The detection limit was 0.1 µg ml -1 .…”

Section: Methodsmentioning

confidence: 99%

Lack of allelopathic effects of the domoic acid-producing marine diatom Pseudo-nitzschia multiseries

Lundholm¹,

Hansen²,

Kotaki³

2005

Mar. Ecol. Prog. Ser.

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Many Pseudo-nitzschia species produce the toxin domoic acid, which accumulates in the food web during blooms, sometimes causing amnesic shellfish poisoning (ASP) in higher trophic levels, including humans. In addition, Pseudo-nitzschia species have been reported to form longlasting monospecific blooms, and a possible explanation for this could be allelopathic effects of the toxin, since domoic acid has been detected in high amounts in the surrounding medium in stationary growth phase. We therefore examined the potential allelopathic effects of P. multiseries and its toxin domoic acid. In mixed-batch culture studies of domoic acid-producing P. multiseries and the algal test species Chrysochromulina ericina, Heterocapsa triquetra, Eutreptiella gymnastica and Rhodomonas marina, no allelopathic effects of P. multiseries were found. Different growth results using 2 different strains of P. multiseries grown with C. ericina were explained by minor differences in pH. Bioassays testing the effect of domoic acid itself on 9 different phytoplankton species, namely C. ericina, E. gymnastica, Karenia mikimotoi, H. triquetra, Heterosigma akashiwo, Prorocentrum minimum, P. micans, Pyramimonas propulsa and R. marina confirmed a lack of allelopathic effects of the toxin. This lack of allelopathic effect of the shellfish-poisoning toxin domoic acid seems to correspond with the results of phytoplankton species causing DSP (diarrheic shellfish poisoning) and PSP (paralytic shellfish poisoning), where it appears that these shellfish-poisoning toxins do not cause allelopathic effects either.

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Section: Methodsmentioning

confidence: 99%

Lack of allelopathic effects of the domoic acid-producing marine diatom Pseudo-nitzschia multiseries

Lundholm¹,

Hansen²,

Kotaki³

2005

Mar. Ecol. Prog. Ser.

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“…This poisoning resulted from consumption of farmed blue mussels Mytilus edulis from Cardigan, Prince Edward Island, Canada that possessed concentrations of DA ranging up to 900 µg g -1 of soft tissue (Quilliam et al 1989, Wright et al 1989. The source of the DA was the diatom (Subba Rao et al 1988, Bates et al 1989) Pseudo-nitzschia multiseries (Hasle 1995) upon which the mussels had been feeding.…”

Section: Introductionmentioning

confidence: 99%

Bacterial involvement in determining domoic acid levels in Pseudo-nitzschia multiseries cultures

Stewart¹

2008

Aquat. Microb. Ecol.

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“…The levels of these low-molecular-weight toxins are typically measured using either bioassay or high-performance liquid chromatography methods (16,23), both of which are time-consuming, relatively low-throughput, and often expensive procedures (11,13). Immunoassays, by contrast, are more suited to highthroughput screening, while being sensitive and highly specific (30), and recent changes in European Union regulations governing the sale of shellfish (8) indicate that immunoassays are becoming more acceptable for shellfish monitoring programs.…”

mentioning

confidence: 99%

Generation of High-Affinity Chicken Single-Chain Fv Antibody Fragments for Measurement of thePseudonitzschia pungensToxin Domoic Acid

Finlay

Shaw

Reilly

et al. 2006

Appl Environ Microbiol

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Antibody-based assay systems are now accepted by regulatory authorities for detection of the toxins produced by phytoplankton that accumulate in shellfish tissues. However, the generation of suitable antibodies for sensitive assay development remains a major challenge. We have examined the potential of using the chicken immune system to generate high-affinity, high-specificity recombinant antibody fragments against phytotoxins. Following immunization of the chicken with domoic acid-bovine serum albumin, a single-chain antibody variable region (scFv) gene library was generated from single V H and V L genes isolated from the immune cells in the spleen and bone marrow. scFvs reacting with domoic acid were isolated by phage display and affinity matured by light chain shuffling, resulting in an approximate 10-fold increase in sensitivity. The isolated scFvs were effectively expressed in Escherichia coli and readily purified by affinity chromatography. They were then used to develop a convenient and sensitive indirect competitive enzyme-linked immunosorbent assay for domoic acid, with a 50% effective dose of 156 ng/ml, which could be used reliably with shellfish extracts. This study demonstrates that chickens provide a valuable model system for the simplified, rapid generation of high-affinity recombinant antibody fragments with specificity for small toxin molecules.The accurate quantification of algal toxins in environmental and food samples is critical for consumer protection. The levels of these low-molecular-weight toxins are typically measured using either bioassay or high-performance liquid chromatography methods (16, 23), both of which are time-consuming, relatively low-throughput, and often expensive procedures (11, 13). Immunoassays, by contrast, are more suited to highthroughput screening, while being sensitive and highly specific (30), and recent changes in European Union regulations governing the sale of shellfish (8) indicate that immunoassays are becoming more acceptable for shellfish monitoring programs.Immunoassay techniques have previously relied on hyperimmune polyclonal sera from rabbits, sheep, or other mammals. These are relatively easy to produce in large quantities but do not offer the consistency required for large-scale application or commercial production. More recently, monoclonal antibodies have been favored because they can be produced in unlimited supply and are more easily standardized. However, monoclonal antibodies have their own set of difficulties, as they are almost exclusively murine in origin (10) and are laborious to screen, and small mammals such as mice do not always provide the high-affinity antibody response to small hapten molecules needed for sensitive assay development (22).The limitations of traditional techniques have led several research groups to investigate the use of phage display to produce antihapten antibodies. The phage display of recombinant antibody libraries is a robust monoclonal antibody technology that is an increasingly attractive method, as it allows the ...

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High-Performance Liquid Chromatography of Domoic Acid, a Marine Neurotoxin, with Application to Shellfish and Plankton

Cited by 145 publications

References 11 publications

Lack of allelopathic effects of the domoic acid-producing marine diatom Pseudo-nitzschia multiseries

Lack of allelopathic effects of the domoic acid-producing marine diatom Pseudo-nitzschia multiseries

Bacterial involvement in determining domoic acid levels in Pseudo-nitzschia multiseries cultures

Generation of High-Affinity Chicken Single-Chain Fv Antibody Fragments for Measurement of thePseudonitzschia pungensToxin Domoic Acid

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