A Temperature-Controlled Fluorescence Fingerprint for Identifying Pseudo-nitzschia hasleana in Harmful Algal Blooms
, Sergey Voznesenskiy 1, Tatiana Dunkai 2,3, Andrei Leonov 1 and Tatiana Orlova 2
Erica L. Seubert
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThere is a large disconnect between the reasons presented for the paper purpose and the results that are given. It is stated in the introduction that this FTC method could be used for HAB monitoring, it is stated in the conclusion that you have shown FTC could be used for HAB monitoring, but there is nothing in between that actually demonstrates this fact. The connection to HABs is solely the fact that a toxic diatom species was chosen as one of the cultures. Why are the other cultures chosen? How would this be beneficial to a monitoring program? Why is this specialized method necessary for monitoring? What does this information do to push us forward in knowing about these microalgae? The paper is failing to communicate to readers the point of this work and why it is something anyone should consider for their own laboratory.
Comments for author File: 
Comments.pdf
The introduction especially has some odd sentences that are not worded well, mudding the communication of the author's point. In the attached file I highlighted some specifically egregious sentences that were incomprehensible.
Author Response
Please see the attachment
Author Response File: Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe study investigates the effect of heating rate on fluorescence temperature curves (FTCs) of toxic Pseudo-nitzschia hasleana, compares it with non-toxic algae, and designs a detection method, finding distinct FTC features of P. hasleana at specific heating rates. It contributes to environmental monitoring by providing a non-invasive optical tool for early harmful algal bloom (HAB) detection and links microalgal physiology to optical signals. This article has certain innovativeness, but it is recommended to undergo the following thorough and sufficient revisions before being accepted.
There are grammatical errors in the manuscript. "heating race" should be corrected to "heating rate"
The abstract lacks specific details about the experimental design. It does not clearly mention the specific range of heating rates used in the study, which is a core experimental condition for readers to quickly grasp the research framework.
The introduction fails to clearly explain why the previous laser-induced fluorescence (LIF) method cannot distinguish between toxic and non-toxic microalgae.
The materials and methods section does not provide detailed components of the culture media used for microalgal cultivation. The specific formulations of f/2 medium (for P. hasleana) and Goldberg medium (for non-toxic algae) are not described, which is critical for other researchers to repeat the experiment.
The results section lacks quantitative data on the changes in normalized fluorescence temperature curves (NFTCs) of P. hasleana under different heating rates.
The discussion does not deeply explore the biochemical mechanism behind the unique fluorescence response of P. hasleana.
In the abstract, the statement "yielding fourfold higher fluorescence intensity at 70-80 °C compared to slower rates" does not clearly specify the comparison object. It is not clear which specific slower heating rate (e.g., 0.5 °C/min or 1 °C/min) the fourfold increase refers to, leading to ambiguity.
The introduction does not mention the specific type of toxin produced by P. hasleana when describing the hazards of HABs. It only generally states that toxic microalgae pose risks to marine ecosystems and human health, but without specifying the toxin type, readers cannot fully understand the specific risks of P. hasleana.
The materials and methods section does not describe the stability control measures for laser power during laser excitation.
Author Response
Please see the attachment
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe argument as to why this paper relates to HABs and why it is a good method to be picked up by plankton monitoring efforts is still weak but has been improved over the last version.
Author Response
Please see the attachment
Author Response File: Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe author has made comprehensive revisions to the article, but there is no content in Figure 1. Besides that, I don't have any further comments.
Author Response
Please see the attachment
Author Response File: Author Response.pdf
