Bacteriota and Antibiotic Resistance in Spiders

Round 1

Reviewer 1 Report

I think the proposed problems have been reasonably modified and improved.

Reviewer 2 Report

Much improved from an earlier version

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

In this study, the microbiota on the surface of spiders collected in chicken farm, slaughterhouse and buildings located in west Slovakia were isolated and identified, and the antibiotic resistance of the microbial species on the surface of spiders was determined. The study demonstrates that 14 spider species carried opportunistic pathogenic  bacteria on their body surfaces that may result in vector-borne transmission of different  pathogens, including zoonoses. Multiresistance and resistance to antimicrobials important for human medicine were recognized in spider isolates. The work could be a contribution in research on microbial compositions of spider body surface and the antimicrobial resistance of their isolates with potential public and environmental health implications. However, there are several main issues that in my opinion should be addressed before this paper can be published:

 

1. The introduction needs to be more focused and the background of the study needs to be laid out much better. In my opinion, the authors should systematically state the research status of endosymbiotic bacteria, intestinal microorganisms and surface microorganisms of spiders, so as to draw the necessity of carrying out the research of spider surface microorganisms. However, from the current introduction part, it is difficult for me to see that the author wants to carry out the relevant research work on spider surface microorganisms. The same case also exists in the abstract.
2. In Table 1, why does the author not mark the collection place; In Table 3, why does the author not list the spider hosts of various microorganisms. If the collection sites of various spiders and the spider hosts of various microorganisms isolated are listed, figure 1-3 will be better understood.
3. There are many languages logic errors in the text: line62, line 72, line 108.
4. There are some mistakes in species taxonomy in this paper: line 64-65.

Reviewer 2 Report

General suggestion
The manuscript shows really interesting results of AMR bacteria isolated from spider species inhabiting Slaughterhouse, Chickens farm and Buildings. But, what is behind the distribution of AMR bacteria find in the different spider species and environments? Despite the important data about antimicrobial resistance, we did not learn nothing about the spider species distribution, even the most frequent (Pholcus phalangioides, Steatoda bipunctata and Steatoda triangulosa), and why this is a health risk as proposed. They are all in the same environment? Which is the risk for humans, animals, etc.? I really think that the manuscript can be improved exploring the correlations among AMR bacteria and spiders species, as well as by exploring the correlations of species and environments (Slaughterhouse, Chickens farm and Buildings). This can be explored by some kinds of networks, but not necessarily. Correlograms can be used. This way, the manuscript will probably reveal some kind of species co-occurrence pattern (bacteria and spiders), that is really difficult to got as is now. Finally, the discussion could be improved based on what should be obtained from the species co-occurrence pattern to be less speculative, and could go a step further of only describe the presence of AMR bacteria.

Most of the Tables and Figures of the current version should be included as Supplemental Material.

Specific suggestions
Title must be changed to "Microbiota and Antibiotic Resistance in spiders"

Intro
lines 59-60 
Suggestion: add a comma
"This phenomena has been referred as hologenomic adaptations, the new properties of microorganisms result from the symbiosis between microorganisms and the hosts they have been inhabiting"

line 62
Suggestion: delete of
"Research on microbiota may reveal potential interaction between the hosts ... "

line 72
Suggestion: that the, ... Review the phrase
"Hu et al. [17] documented the gut microbial profiling and observed that the, ... "

Material & Methods
2.1. Sample preparation
line 98
Suggestion: identify and better describe slaughterhouse, buildings and chicken farms with names, georeferentiation (lat & lon), etc. It is important to have the possibility of compare what was identified here with other samples as well with samples of the same place in other times.

line 106-108
Suggestion: review the phrase
"Tripton Soya agar (TSA), Tripton Sugar Iron agar (TSI), Anaerobic agar (AA), and 106 Blood agar (BA) (Sigma-Aldrich® , St. Louis, USA) were used for detection of microbial 107 count, 
Enterobacteriacea, anaerobic microbiota and unspecified microorganisms. differ- 108 ent spectra of bacteria."

Results
3.1. Qualitative analysis of isolated microbiota from spiders
Suggestion: Species names in Italics

Table 2.
Suggestion: spiders species names simplified, example M. ferruginea without typing (Panzer, 1804) that already is in Table 1 will improve table legibility.

Reviewer 3 Report

The manuscript entitled “Microbiota and Antibiotic Resistance in Arthropods” by Kačániová et al. describes the different bacterial culturing approaches, isolation, and identification of bacteria.  Also, done with antibiotic testing with cultured bacteria.  Although the author's carried out extensive work, almost all the section manuscript was poorly written.  In my view, it requires extensive editorial work.  See some of the specific comments below:

The title needs to be more specific; Arthropods is too broad.

“Introduction” section was narrowly written; much of the relevant existing literature is not mentioned; for example, see the Kumar et al. 2020, https://doi.org/10.3389/fevo.2020.588102; Keiser et al 2019, https://doi.org/10.1098/rsbl.2019.0436 etc.

Line:  “Sample was diluted” from the external or internal sample for the spider?

Line 97:  “slaughterhouse, buildings and chicken farms” I strongly recommend adding more information about the locations, year of the sampling, specific location, etc.

Line 100:  “Supplementary file Table 1” I don’t see any data in the manuscript.

Line 101:  “frozen at −20 °C for 5 min” keeping at freezing temperature also kill the bacteria.

Line 102:  “1 mL of sterile 0.87% (w/v) NaCl” solution used for culture was only from the surface or only from internal? 

Line 103: “microbial” change it to “bacterial”.  The medium used in these studies was designed to culture the bacteria.

Table 1.  I also suggested adding the source of the Spider was obtained.

Line 108:  “unspecified microorganisms” define as?

Line 108:  “different spectra of bacteria”  incomplete sentences

Lines 106 to 107: Authors used Tripton Sugar Iron agar (TSI), Anaerobic agar (AA), and Blood agar (BA) was used culture, whereas identification only TSA.  Are all the bacteria cultured from other mediums also able to on TSA?

Line 113: “three to five bacterial colonies ” on what bases were these colonies selected?

Line 130:  how many “isolated species”?

Table 2: The “log cfu/g” how did the author calculate without measuring the spiders weight?

“3.1.  Qualitative analysis of isolated microbiota from spiders” scientific name in this section need to be italicized.

The result in “Table 2” is not clear how the t-test was done using multiple mediums and different spiders?   

Line 158: “Totally, 28 genera and 56 microbial” was these from all the medium?  Was all the colony isolated and identified?