Harmful algal bloom (HAB) species include members of several phytoplankton taxa that cause a variety of negative effects on aquatic ecosystems, human health, and economies, such as trophic accumulation of toxins, shellfish poisoning, fishery and beach closures, and others (e.g., Smayda 1997; Scholin et al. 2000;Hoagland and Scatasta 2006). Light microscopy has been traditionally used to identify and enumerate HAB species, but this approach is time consuming and accurate species identification is difficult among morphologically similar taxa and when the organism of interest is rare within a complex microbial assemblage (Anderson 1995; Anderson et al. 2005). Consequently, there has been a recent surge in the development of methods that enable rapid and accurate detection and quantification of HAB species and the substances they produce to augment research and management activities that facilitate early warning of bloom events to close fisheries or alert health officials. Examples include antibody tests, (e.g., Campbell et al. 1989;Shapiro et al. 1989), optical detection (e.g., Kirkpatrick et al. 2000;Robbins et al. 2006), molecular approaches (e.g., Anderson 1995;Groben et al. 2004), and observatory platforms (e.g., Sosik et al. 2003;See et al. 2005;Olson and Sosik 2007;Paul et al. 2007;Roman et al. 2007; Scholin et al. 2009). Despite these numerous advances, comparisons between multiple approaches are surprisingly rare. Godhe et al. (2007) describes a novel workshop that entailed cross-comparisons of methods for quantifying the "red tide" dinoflagellate Alexandrium tamarense. While results elucidated the relative performance of several techniques, varying culture conditions and use of shared equipment prevented direct comparisons, highlighting the importance for a rigorous sideby-side methods comparison accomplished by splitting a single sample followed by analyses with multiple methods.
Comparison
AbstractIn this study, we directly compared two molecular techniques, sandwich hybridization assay (SHA) and quantitative PCR (qPCR), for quantifying laboratory cultures of the ichthyotoxic raphidophyte Heterosigma akashiwo. To maximize comparisons, all experiments entailed raising H. akashiwo in the laboratory, generating cellular homogenates then splitting them such that for each sample, the same homogenate was analyzed using both SHA and qPCR. To verify molecular data, all experiments included cell counts using light microscopy and a Sedgewick Rafter chamber. Results of standard curves for several geographically distinct strains using SHA were similar to prior research whereas others exhibited significantly different slopes, suggesting physiological differences between isolates. Data generated by qPCR showed a high degree of correlation with SHA responses. We also investigated the use of each method for quantifying H. akashiwo samples preserved with Lugol's iodine solution. Neither SHA nor qPCR produced results that were significantly different from initial values for Lugol's preserved samples stored under refri...