Occurrence of four species of algae in the marine water of Hong Kong.

Harmful algal blooms (HABs) have broken out frequently throughout the world in recent decades; they are caused by the rapid multiplication of algal cells in near-coastal waters polluted with nitrogen and phosphorus and greatly affect the quality of marine water and human health. Over the past several decades, climate change and increasing environmental degradation have provided favourable growth conditions for certain phytoplankton species. Therefore, it is essential to rapidly identify and enumerate harmful marine algae to control these species. In this study, quantitative PCR (qPCR) was used to detect four representative species of HABs that are widespread in the marine water of Hong Kong, namely, Alexandrium catenella, Pseudo-nitzschia spp., Karenia mikimotoi and Heterosigma akashiwo. We applied qPCR with the dye SYBR Green to detect Alexandrium spp. and Pseudo-nitzschia spp. and used TaqMan probe for the enumeration of Karenia mikimotoi and Heterosigma akashiwo. The total genomic DNA of these algae from Hong Kong marine water was extracted successfully using the CTAB method, and for each kind of alga, we constructed a ten-fold series of recombinant plasmid solutions containing certain gene fragments of 18S rDNA and ITS1-5.8S-ITS2 as standard samples. Ten-fold dilutions of the DNA of known numbers of the extracted algal cells were also used to create an additional standard curve. In this way, the relationship between the cell number and the related plasmid copy number was established. The qPCR assay displayed high sensitivity in monitoring marine water samples in which the low concentrations of harmful algae were not detected accurately by traditional methods. The results showed that the cell numbers of the four species were all in low abundance. For Alexandrium catenella, the cell abundances at 12 sites ranged from 3.8×102 to 4.3×103 cellsL-1 , while H. akashiwo, K. mikimotoi and Pseudo-nitzschia ranged from 1.1×102 to 1.3×103 , from 23 to 6.5×102 and from 45 to 3.3×103 cellsL-1 , respectively. The concentrations of these algae were much lower than those observed during outbreaks of HABs in Hong Kong. These results may be useful for local aquaculture development and may provide effective suggestions and a theoretical basis for HAB monitoring and management.