Rapid Enzyme-linked Immunosorbent Assay for Detection of the Algal Toxin Domoic Acid

Litaker, R. Wayne; Stewart, Thomas N.; Eberhart, Bich-Thuy L.; Wekell, John C.; Trainer, Vera L.; Kudela, Raphael M.; Miller, Peter E.; Roberts, A; Hertz, Cassandra; Johnson, Tyler A.; Frankfurter, Greg; Smith, G. Jason; Schnetzer, Astrid; Schumacker, Joe; Bastian, Jonnette L.; Odell, Anthony; Gentien, Patrick; Gal, Dominique Le; Hardison, D. Ransom; Tester, Patricia A.

doi:10.2983/0730-8000-27.5.1301

Cited by 42 publications

(24 citation statements)

References 32 publications

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“…Cultures were then filtered with 25 mm Whatman GF/C filters, and stored in À 80 1C until processing. DA was extracted by adding 1.5 ml 20% methanol, sonicated on ice for 3 min, centrifuged for 10 mins at 3000 g and the supernatant filtered with a 0.2 mm syringe filter (Litaker et al, 2008). DA concentration was measured on a microplate reader (Spectromax M2, Molecular Devices, Sunnyvale, CA, USA) following the procedure described in Mercury Science DA kit (Mercury Science, Durham, NC, USA).…”

Section: Da Measurement In Binary Culturementioning

confidence: 99%

Host-specific adaptation governs the interaction of the marine diatom, Pseudo-nitzschia and their microbiota

et al. 2013

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The association of phytoplankton with bacteria is ubiquitous in nature and the bacteria that associate with different phytoplankton species are very diverse. The influence of these bacteria in the physiology and ecology of the host and the evolutionary forces that shape the relationship are still not understood. In this study, we used the Pseudo-nitzschia-microbiota association to determine (1) if algal species with distinct domoic acid (DA) production are selection factors that structures the bacterial community, (2) if host-specificity and co-adaptation govern the association, (3) the functional roles of isolated member of microbiota on diatom-hosts fitness and (4) the influence of microbiota in changing the phenotype of the diatom hosts with regards to toxin production. Analysis of the pyrosequencing-derived 16S rDNA data suggests that the three tested species of Pseudo-nitzschia, which vary in toxin production, have phylogenetically distinct bacterial communities, and toxic Pseudo-nitzschia have lower microbial diversity than non-toxic Pseudonitzschia. Transplant experiments showed that isolated members of the microbiota are mutualistic to their native hosts but some are commensal or parasitic to foreign hosts, hinting at co-evolution between partners. Moreover, Pseudo-nitzschia host can gain protection from algalytic bacteria by maintaining association with its microbiota. Pseudo-nitzschia also exhibit different phenotypic expression with regards to DA production, and this depends on the bacterial species with which the host associates. Hence, the influences of the microbiota on diatom host physiology should be considered when studying the biology and ecology of marine diatoms.

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Section: Da Measurement In Binary Culturementioning

confidence: 99%

Host-specific adaptation governs the interaction of the marine diatom, Pseudo-nitzschia and their microbiota

et al. 2013

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“…An example of an antibody test is the Jellett test kit [5], which indicates the presence of high levels of a toxin. A more advanced test kit is one produced by Mercury Science, Inc. that tests for several levels of domoic acid concentration [6]. This approach offers considerable value for monitoring programs by quickly identifying both samples that are above regulatory limits and those that require more precise testing.…”

Section: Rapid Detection Methodsmentioning

confidence: 99%

Integration of Data for Nowcasting of Harmful Algal Blooms

Stumpf

Fleming-Lehtinen²,

Granéli

2010

Proceedings of OceanObs'09: Sustained Ocean Observations and Information for Society

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Harmful algal blooms (HABs) are a significant and potentially expanding problem around the world. Resource management and public health protection require sufficient information to reduce the impacts of HABs by response strategies and through warnings and advisories. To be effective, these programs can best be served by an integration of improved detection methods with both evolving monitoring systems and new communications capabilities. Data sets are typically collected from a variety of sources, these can be considered as several types: point data, such as water samples; transects, such as from shipboard continuous sampling; and synoptic, such as from satellite imagery. Generation of a field of the HAB distribution requires all of these sampling approaches. This means that the data sets need to be interpreted and analyzed with each other to create the field or distribution of the HAB. The HAB field is also a necessary input into models that forecast blooms. Several systems have developed strategies that demonstrate these approaches. These range from data sets collected at key sites, such as swimming beaches, to automated collection systems, to integration of interpreted satellite data. Improved data collection, particularly in speed and cost, will be one of the advances of the next few years. Methods to improve creation of the HAB field from the variety of data types will be necessary for routine nowcasting and forecasting of HABs.

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“…DA was measured in fish viscera using 2 methods: HPLC and Rapid ELISA (Mercury Science). Both methods are comparable, but ELISA is a more desirable method due to its lower LOD and faster, more cost-effective procedure (Litaker et al 2008). DA was measured with ELISA in the following fish samples: (1) all muscle tissue samples, except for anchovy collected in 2007, when DA in muscle tissue was measured using HPLC; (2) viscera of 6 sardine specimens examined in the DA degradation and diffusion experiment; and (3) fish collected in May 2009.…”

Section: Methodsmentioning

confidence: 99%