Efficacy of Phosphine on Different Life Stages of Alphitobius diaperinus and Tenebrio molitor (Coleoptera: Tenebrionidae)
Round 1
Reviewer 1 Report
Please provide a few words on LT50, LT95, and LT99. What are they?
What is the origin of the phosphine that was used in this study?
In Bioassay I, why didn't the investigators consider studying if the phosphine concentration was below 50 ppm?
In Bioassay III, why didn’t the investigators consider studying the phosphine concentration below 3000 ppm?
Did the authors observe any morphological changes in the mealworms? decreased body size, etc.
Did the authors consider testing the phosphine resistance of individuals who survived the first phosphine exposure? There may be enhanced resistivity to phosphine for the second generation.
The authors forgot to add a conclusions section.
Author Response
The authors would like to thank the reviewer for the comments and recommendations on improving the quality of the manuscript before resubmission. A revised version is submitted with the proposed corrections/additions addressed. A point by point reply on each comment/correction can be found below.
Please provide a few words on LT50, LT95, and LT99. What are they?
REPLY: Done as suggested. Please see line 145. LT50, LT95 and LT99 stand for the lethal time for killing 50, 95, and 99%of the exposed individuals. Please see the attachment.
What is the origin of the phosphine that was used in this study?
REPLY: The phosphine production conducted in the lab as described by Agrafioti et al. 2019 the by using the Phosphine Tolerance Test (PTT, Detia Degesch GmbH, Germany. See line 13 and line 117.
In Bioassay I, why didn't the investigators consider studying if the phosphine concentration was below 50 ppm?
REPLY: Since exposure to 50 ppm, didn’t lead to complete mortality of eggs, we considered testing a higher, and not a lower concentration. That’s why we also evaluated the exposure at 100 ppm, which was adequate for killing all life stages tested, and not higher concentrations. Exposure to lower concentration than 50 ppm for 3 days do not lead to complete mortality and higher than 100 ppm is not necessary.
In Bioassay III, why didn’t the investigators consider studying the phosphine concentration below 3000 ppm?
REPLY: This bioassay is based on a standard resistant screening protocol by Detia Degesch GmbH, Germany. Standard resisting protocols are used in studies evaluating the effect of phosphine, so that the results are comparable with those of other studies on the same or different species (Agrafioti et al., 2019, Aulicky et al., 2019, Athanassiou et al., 2019).
Did the authors observe any morphological changes in the mealworms? decreased body size, etc.
REPLY: No morphological changes were observed in living insects. However, apparently the appearance of the dead was different from that of the living.
Did the authors consider testing the phosphine resistance of individuals who survived the first phosphine exposure? There may be enhanced resistivity to phosphine for the second generation.
REPLY: Testing the phosphine resistance of individuals who survived the first phosphine exposure is a very interesting and important issue. However, this is not included in the objectives of this particular investigation. In addition, as the number of insects survived was very low, conducting such an experiment would have been impossible. The authors consider investigating this effect in subsequent research by using protocols aiming this particular issue.
The authors forgot to add a conclusions section.
REPLY: according to the journal's guidelines, a conclusion section is not necessary
Reviewer 2 Report
The authors need to add stattistical analysis error bar in the Figure 1 and 2.
over all writign of the study goals and results are good.
Author Response
Reviewer 2: The authors need to add stattistical analysis error bar in the Figure 1 and 2.over all writign of the study goals and results are good.
REPLY: The authors would like to thank the reviewer for the comments and recommendations on improving the quality of the manuscript before resubmission. A revised version is submitted with the proposed corrections/additions addressed. A point by point reply on each comment/correction can be found below.
On the bars where the mortality is 100%, the statistical error is 0, and it is not visible in the Figures. On the bars representing different percentages you can see the standard error bars.
Reviewer 3 Report
Dear authors
Please see my comments in the attached file.
Regards
Comments for author File: 
Comments.pdf
Author Response
Reviewer 3:
Dear authors
Please see my comments in the attached file.
Regards
REPLΥ: The authors would like to thank the reviewer for the comments and recommendations on improving the quality of the manuscript before resubmission. A revised version is submitted with the proposed corrections/additions addressed. A point by point reply on each comment/correction can be found below.
All the suggested changes have been incorporated in the manuscript.
