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Fisheries and Oceans Pêches et Océans Pacific Region August 30, 2016 Dr. Michael V. K. Sukhdeo Editor, Journal of Parasitology Dept. Ecology, Evolution and Notural Resources Rutgers University New Brunswick, New Jersey 08901 Dear Dr. Sukhdeo: Subject: Journal of Parasitology MS 16-24R1 Please find enclosed a revised manuscript (and accompanying files) entitled: “Two novel microsporidia in skeletal muscle of pike-perch Sander lucioperca and burbot Lota lota in Finland”. We have further modified the text as suggested by comments from the editor and a reviewer, and as itemized on the following pages. On behalf of my co-authors, I would like to acknowledge the effort made by the editorial staff and reviewers to improve this manuscript. Sincerely yours, Simon R.M. Jones Pacific Biological Station -2- Associate Editor comments: The revised version of the manuscript addressed most of the reviewer’s concerns and is much improved. I agree with the reviewer that making it very clear the measurements were based on TEM is important in the abstract, methods and results. In the abstract, this could go at the end of the sentence on line 11 (“...wide, based on transmission electron microscopy sections.”). In the results, “.., based on TEM” can follow the reported measurements on lines 119 and169. Changes have been made as suggested (lines 10, 11, 18, 118, 121, 167) I also agree with the reviewer’s second comment that some broader ecological context would benefit the paper. In the last paragraph, the impacts of other microsporidia are mentioned. What impacts, if any, would be expected from these new species? In the Intro, the authors mention that microsporidian infections can degrade host musculature. Is that expected here? The TEM work revealed some host response. Would that lead to tissue damage? Just a little more context would be good to bring this all together. A broader ecological context is now discussed in the last paragraph (line 249 – 266). The sentence refered to in the introduction has been relocated to this part of the discussion. Reviewer #2: The manuscript is significantly improved following this revision and the authors have addressed the majority of comments. I only have two minor comments for the manuscript. One comment is again in regards to the spore sizes being based on TEM measurements. The authors have stated in the Materials and Methods under the electron microscopy section that microsporidian dimensions were made from ultrathin sections viewed in electron micrographs. This can easily be overlooked when reading the MS. It is known that sizes of spores are affected by tissue processing, particularly TEM processing. Considering that spore size is an important morphological criterion for microsporidia and that future readers of this paper will be comparing the sizes of different microsporidian species, it would benefit to clearly state that measurements were from TEM directly when the measurements are stated. This is particularly important in the abstract since the measurements are stated with no reference to this. It would be important to simply write "based on measurements made from TEM" after stating the measurements for each species in the abstract. This will be very helpful to future taxonomists reviewing the morphology of these new species. Additionally authors should consider stating this in the Diagnosis section after mention of spore dimensions in the species description. Changes have been made (see above) -3- The second comment is in reference to the ecological context of the parasites in these fish species. Journal of Parasitology is avoiding direct species descriptions unless they have an important ecological context. The authors have added a few sentences in the Introduction to remedy this, including some catch numbers of both fish species and that parasite infections can degrade appearance of the musculature. This helps, but discussing the ecological context beyond stating catch numbers would strongly benefit the manuscript. The Discussion section includes only morphological aspects of the microsporidia. It would benefit to include a paragraph discussing the ecological context. For example, based on the results and infection intensities do the authors believe that these parasites will have an impact on the musculature of the fish (from the perspective of fisherman)? Health of the fish? Do closely related microsporidian species cause problems in musculature of the fish? Heterosporis is known to cause unsightly lesions in the musculature making the filets of heavily infected fish unusable. Have fisherman or others noticed these infections as problematic in the fish? Maybe compare these new species to other parasites from this perspective. Also any information on biology of the fish, transmission dynamics of microsporidia, and how these may interact. These are just some ideas on what could be included from an ecological perspective to help discuss the importance of these parasites. A paragraph describing a the broader ecological context of our findings has been included in the discussion