Chattonella species, C. marina and C. ovata, are harmful raphidophycean flagellates known to have hemolytic effects on many marine organisms and resulting in massive ecological damage worldwide. However, knowledge of the toxigenic mechanism of these ichthyotoxic flagellates is still limited. Light was reported to be responsible for the hemolytic activity (HA) of Chattonella species. Therefore, the response of photoprotective, photosynthetic accessory pigments, the photosystem II (PSII) electron transport chain, as well as HA were investigated in non-axenic C. marina and C. ovata cultures under variable environmental conditions (light, iron and addition of photosynthetic inhibitors). HA and hydrogen peroxide (H2O2) were quantified using erythrocytes and pHPA assay. Results confirmed that% HA of Chattonella was initiated by light, but was not always elicited during cell division. Exponential growth of C. marina and C. ovata under the light over 100 µmol m−2 s−1 or iron-sufficient conditions elicited high hemolytic activity. Inhibitors of PSII reduced the HA of C. marina, but had no effect on C. ovata. The toxicological response indicated that HA in Chattonella was not associated with the photoprotective system, i.e., xanthophyll cycle and regulation of reactive oxygen species, nor the PSII electron transport chain, but most likely occurred during energy transport through the light-harvesting antenna pigments. A positive, highly significant relationship between HA and chlorophyll (chl) biosynthesis pigments, especially chl c2 and chl a, in both species, indicated that hemolytic toxin may be generated during electron/energy transfer through the chl c2 biosynthesis pathway.

In this paper, an algal identification and concentration determination method based on discrete excitation fluorescence spectra is proposed for online algae identification and concentration prediction. The discrete excitation fluorescence spectra of eight species of harmful algae from four algal categories were assessed. After determining typical excitation wavelengths according to the distribution of photosynthetic pigments and eliminating strongly correlated wavelengths by applying the hierarchical clustering, seven characteristic excitation wavelengths (405, 435, 470, 490, 535, 555, and 590 nm) were selected. By adding the ratios between feature points (435 and 470 nm, 470 and 490 nm, as well as 535 and 555 nm), standard feature spectra were established for classification. The classification accuracy in pure samples exceeded 95%, and that of dominant algae species in a mixed sample was 77.4%. Prediction of algae concentration was achieved by establishing linear regression models between fluorescence intensity at seven characteristic excitation wavelengths and concentrations. All models performed better at low concentrations, not exceeding the threshold concentration of red tide algae outbreak, which provides a proximate cell density of dominant algal species.

Summary Phaeocystis globosa frequently proliferates in eutrophic waters and forms ichthyotoxic algal blooms that cause massive fish mortalities in marine ecosystems. One of the ichthyotoxic metabolites was identified as the glycolipid‐like hemolytic toxin, reported to be initiated under light conditions. However, the association between hemolytic activity (HA) and photosynthesis of P. globosa remained unclear. Light spectra (blue, red, green, and white) and 3‐(3,4‐dichlorophenyl)‐1,1‐dimethylurea (DCMU) were selected as the stressors to stimulate the hemolytic response of P. globosa in relation to the light and dark photosynthesis reaction. Hemolytic activity in P. globosa was sensitive to the light spectrum as it decreased from 93% to nearly undetectable (1.6%) within 10 min of transfer from red (630 nm) to green light (520 nm). This indicates that the vertical transformation of P. globosa from deep to surface waters (dominated by green light and all light spectra, respectively) may drive the hemolytic response in coastal waters. However, regulation of photosynthetic electron transfer in the light reaction of P. globosa was excluded by the evidence of inconsistent response of HA to photosynthetic activity. The biosynthesis of HA may interfere with the pathway of photopigments diadinoxanthin or fucoxanthin, and the metabolism of three‐ and five‐carbon sugars (GAP and Ru5P, respectively), which ultimately lead to changes in the alga's hemolytic carbohydrate metabolism.