A New Species of the Medically Important Scorpion Genus, Hemiscorpius Peters, 1861 (Hemiscorpiidae), from Southern Iran

The manuscript of Amiri et al. is an interesting contribution in a group of scorpions of medical importance, therefore I consider it valuable and worth publishing after solving some minor problems.

The manuscript is in general well written, images are of excellent quality, and the description is complete and made according to modern standards.

Many thanks

I have only some suggestions regarding the diagnosis, the description of the ecology of the species and some minor problems with the description.

Thanks for your valuable comments

The diagnosis should be improved, when authors refer that this species is larger than H. acanthocercus authors should provide more specific data about it: range of variation of both species, number of studied specimens in each case, mean, differences between genders. This applies to any other measurement in the diagnosis, in the description and in the intraspecific variation.

Details are provided in Table 2 and also in Reference 4, which is cited. To avoid repetition, details are not listed here. The sentence was edited.

Authors refer to differences in total length of metasoma in males and females, but they do not provide ranges of variation, nor ratios in each case to support this statement.

Edited

The first paragraph of the results (3.1. morphology, ecology and distribution) should be omitted, since the information provided in it is too vague and not connected, and should be expanded in other parts of the manuscript.

This section removed

I strongly suggest including the description of the hemispermatophore of this species, since this is a structure that has proved to be of highly diagnostic value in other scorpion species.

This is a separate and time-consuming task that can be done to compare different genera or species. It is not possible at this time.

Authors should provide more data about the ecology of this species, the environment where in was collected, the range of altitudes, the type of vegetation, the period of the year, etc.

Added in discussion

Authors mention two species groups in Iran, with differences in their trichobothrial pattern; are these the only morphological differences? can these groups be part of different subgenera or monophyletic species groups? are these differences also present in other species outside Iran?

We have indicated that this question remains to be tested by phylogenetic analysis with a broader sample of species (including species from beyond the borders of Iran).

In figure 3 the specimens could be better positioned.

Because the samples were stored in 96% ethanol, they dried out and remained in this state. Photographing and editing pictures takes a lot of time, this is not currently possible.

I have included other minor suggestions in the manuscript.

The text was corrected based on your comments

please provide some kind of measure of these differences, it could be a ratio respect to the total length, or the range of variaton in each gender. Also include range of variation in size and complete the data of pectinal teeth with number of studied specimens and the median.

Some details added. Details for number of studied specimens are provided in Table 2.

In the reviewed MS a group of scientists from Iran leaded by a world scorpion expert from the U.S. give their very interesting results on morphological and genetic diversity of the scorpions of the genus Hemiscorpius collected along the southern border of Iran and in adjacent areas. The authors provided high quality detailed morphological description of the new species and performed a series of molecular phylogenetic analyses focused on phylogeny of Hemiscorpius. The MS is well-written, however some parts of the MS need clarifications, e.g. data on geography, interpretation of the COI tree, genetic diversity of H.leptus and other Hemiscorpius species analyzed. Tables and Figures need minor revision. Some additional remarks are given below.

Thanks for your valuable comments

Abstract is very short. Please, give some more details on the phylogenetic analyses and what can be concluded from those analyses. Please revise the sentence on the geographic barrier: it is unprepared, sounds very short and poorly linked with other part of the Abstract

More details were added

There are no line numbers

The MS is formatted according to the journal template and does not include line numbers.

Intro, paragraph 1: please add brief general information about the phylogenetic position and phylogenetic relations of Hemiscorpius among Scorpionida based on currently published literature.

Added

Intro, last paragraph: the Jebal Barez Mountains appear to have provided a geographical barrier, separating the new species from its closest relative – 1) this is a hypothesis, it is inappropriate to give in in Introduction, it is better to include this in the Discussion 2) please, indicate the mountains in the Fig. 1. According to what is seen in the Fig. 1, the mountains are in the left and there are no distinct mountainous barrier between inverted blue triangle and green stars. So, what barrier is actually mentioned? Please, explain and demonstrate this unambiguously.

Moved to discussion

Fig.1, additional remark. Only type localities are shoun in Fig.1 Could you please show all findings of different species of Hemiscorpius in Iran so that their distribution areas were more clear?

A comprehensive study has not been conducted for all species of this genus in Iran. Also, some of the data on the distribution of the various species are unreliable (there are many misidentifications). For this reason, only the type localities of each species are mapped.

with citations for 10,000 iterations – please, check if this is correct

Its correct

https://phylo.org – please check if you referenced CIPRES correctly

Edited

mrBayes – please, give ref for this software and for all other soft used in the study

Added

3.2. Molecular Systematics The new COI gene sequences were 624 bp long, containing 438 conserved (70.19%),186 variable (29.81%), and 117 parsimony informative (18.75%) sites. – please, add information on diversity of the aminoacid differences. Are all substitutions synonymous or not? How many aminoacid positions differ among different Hemiscorpius species?

This is not common for articles on this group of animals, and we don’t think it is necessary for this study.

Fig.2. Please add geography information in the phylogenetic tree. Please, comment the COI cladogram in terms of geography: do COI clades and COI divergence coincide/contradict geographical distributions of the Hemiscorpius species?

The focus of this research is on the description of the new species. On the other hand, there is no sequence of all the species present in the region to explain the geographical events and speciation of this genus in the Iranian Plateau.

Fig.2. Please, explain the codes used in the tip names

Edited

Table 1. Please, revise this Table. It appears that you have samples of. H.lepturus from Iraq and iran. Please explain this in Material and Methods. Also, revise the column names in the Table.

We only sequenced one sample for the new species. Other sequences (as mentioned in the Materials and Methods section) were obtained from GenBank.

Table 1. It looks like for some species (H. lepturus) your data indeed may correctly reflect the diversity within species since you have analyzed remote populations, but for other species (e.g. the new one) you refer to genetic diversity within one population, because all specimens were collected very close to each other, aren’t they? Please, explain this better to avoid misunderstanding

There are only two sequences of H. lepturus, one from Iran and the other from Iraq. Therefore, intraspecific genetic diversity cannot be calculated for them. The minimum genetic distance between the two species is 0.089. It seems that the minimum difference between species is about 0.80 and within species is 0.040.

Etymology. Please, specify gender and part of speech of the new species name

Added

Did you register the new species in Zoobank?

Added in the text

edipalps: Femur elongate, 2.86 times longer than wide - pedipalps

Edited

Two species groups of Hemiscorpius… - please, indicate these groups in the COI phylogeny and map the morphological characters separating these groups on the tree. Does your COI phylogeny support these groups or not? Please, mention this in the discussion.

In this study, the sequence is available from only one species from the first group, which is located at the base of the tree. Given the small number of sequences for all species, this cannot be discussed with certainty.

H. lepturus (both in Results and Discussion) – please explain all what you wish to say about this species in more detail. Currently it is presented suboptimal in the MS, however it is very interesting.

The focus of this study was on describing the new species. However, since only one sequence was available from each of the two Iranian and Iraqi populations of H. lepturus, it was not possible to investigate further.