An in vivo fluorescence discrimination technique for phytoplankton population was developed by using Wavelet packet transform, cluster analysis and non-negative least squares. The technique was used to analyze water samples from different sea regions. For simulative mixtures, when dominant species account for 60%, 70%, 80%, 90% at the division level, the correct discrimination ratios (CDRs) are 83.0%, 99.1%, 99.7% and 99.9% with the relative contents of 58.5%, 68.4%, 77.7% and 86.3%, respectively; when the algae dominance are 60%, 70%, 80%, 90%, 100% at the genus level, the CDRs are 86.1%, 94.9%, 95.2%, 96.8% and 96.7%, respectively. For laboratory mixtures, the CDRs are 88.1% and 78.4% at the division and genus level, respectively. For field samples, the CDRs were 91.7% and 80% at the division and genus level, respectively (mesocosm experiments), and the CDRs were 100% and 66.7% at the division and genus level, respectively (Jiaozhou Bay). The fluorometric technique was used to estimate the phytoplankton community composition and relative abundance of different classes for the April 2010 cruise in the Yellow Sea with the results agreeing with those in published papers by other authors.
DUAN YaliSU RongguoSHI XiaoyongWANG XiulinZHU ChenjianSUN Yan
The discrete excitation-emission-matrix fluorescence spectra (EEMS) at 12 excitation wavelengths (400, 430, 450, 460, 470, 490, 500, 510, 525, 550, 570, and 590 nm) and emission wavelengths ranging from 600-750 nm were determined for 43 phytoplankton species. A two-rank fluorescence spectra database was established by wavelet analysis and a fluorometric discrimination technique for determining phytoplankton population was developed. For laboratory simulatively mixed samples, the samples mixed from 43 algal species (the algae of one division accounted for 25%, 50%, 75%, 85%, and 100% of the gross biomass, respectively), the average discrimination rates at the level of division were 65.0%, 87.5%, 98.6%, 99.0%, and 99.1%, with average relative contents of 18.9%, 44.5%, 68.9%, 73.4%, and 82.9%, respectively; the samples mixed from 32 red tide algal species (the dominant species accounted for 60%, 70%, 80%, 90%, and 100% of the gross biomass, respectively), the average correct discrimination rates of the dominant species at the level of genus were 63.3%, 74.2%, 78.8%, 83.4%, and 79.4%, respectively. For the 81 laboratory mixed samples with the dominant species accounting for 75% of the gross biomass (chlorophyll), the discrimination rates of the dominant species were 95.1% and 72.8% at the level of division and genus, respectively. For the 12 samples collected from the mesocosm experiment in Maidao Bay of Qingdao in August 2007, the dominant species of the 11 samples were recognized at the division level and the dominant species of four of the five samples in which the dominant species accounted for more than 80% of the gross biomass were discriminated at the genus level; for the 12 samples obtained from Jiaozhou Bay in August 2007, the dominant species of all the 12 samples were recognized at the division level. The technique can be directly applied to fluorescence spectrophotometers and to the developing of an in situ algae fluorescence auto-analyzer for phytoplankton population.
The 3D fluorescence discrimination of phytoplankton classes was investigated by SA4 multiwavelet,GHM multiwavelet,and coifman-2(coif2) wavelet analysis.Belonging to 35 genera of 7 major phytoplankton divisions in the inshore area of China Sea,Single species cultures of 51 phytoplankton species were employed.The second scale vector (Ca2) of SA4,Ca2 of GHM and the third scale vector (Ca3) of coif2 were selected as feature spectra by Bayesian discriminate analysis (BDA).The reference spectra were obtained via hierarchical cluster analysis (HCA).With average high correct discrimination ratios (CDRs),reference spectra were representative to phytoplankton species.For one-algae samples,the average CDRs were 95.6% at genus level and 86.7% at division level.For the laboratory mixed samples,the average CDRs (one division accounted for 25%,75% or 100% of the total biomass) were 86.6%,91.4% and 100% at division level.Moreover,the average CDRs of the dominant species (accounted for 75%) was 79.8% at genus level.Results for the in situ samples were coincided with the microscopic ones at division level with the relative contents of 54.3%-96.5%.The fluorometric discriminating technique was further tested during the cruise in Bohai Sea recently.
