Molecular characterization of enteroviruses circulating among pigs and goats in two Central African countries, Cameroon and the Central African Republic
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To date, data on animal enteroviruses (EVs) are scarce, especially in Central Africa. The aim of this study was to characterize EVs among pigs and goats in Cameroon and the Central African Republic (CAR). A total of 226 pig and goat faecal samples collected in two previous studies carried out in Cameroon and CAR were pooled and screened with molecular assays targeting EV-Es, EV-Fs and EV-Gs. EV genomes were amplified by RT-PCR and their sequences were obtained by Illumina sequencing and de novo assembly. Based on the capsid sequences, 27 EV-G sequences were identified and assigned to 11 virus types, while no EV-E or EV-F was observed. Phylogenetic analysis revealed that the EV-Gs detected in Central Africa do not form specific clusters compared to EV-Gs previously reported in other continents. This suggests a worldwide circulation of EV-Gs, which is likely due to the massive international trade of live animals. One human EV, EV-C99, which belongs to the species Enterovirus C , was detected in pigs. This is the third detection of such an event in a similar context, reinforcing the hypothesis that some EV-Cs could be infecting pigs. Our work provides new data on the genetic diversity of EVs circulating among domestic animals in Central Africa.
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Thank you very much for submitting your revised manuscript to Access Microbiology and applying the reviewers' suggested corrections. I am pleased to let you know that it has now been accepted for publication. Congratulations!
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Thank you for submitting your manuscript for publication in Access Microbiology. It has been examined by expert reviewers who have concluded that the work is of interest to the readership of Access Microbiology. However, a minor amendment of this manuscript will be required before a decision can be made on its publication. I will be pleased to consider a revised manuscript along with a document including a point by point response to each of the reviewers comments. Your revised manuscript may be returned to one or more of the original reviewers, along with your itemised response to the reviewers’ comments.
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Comments to Author
1. Methodological rigour, reproducibility and availability of underlying data The methodology is clearly detailed. The authors provide all the information necessary to ensure the reproducibility of the analyses. 2. Presentation of results The results are well presented. The positives obtained were confirmed by Illumina technology. The authors made a phylogenetic tree from the entire VP1 gene, two complete genomes of EV-G were obtained, the divergence of EV-Gs is well demonstrated in figure 1. These results are robust and clearly presented in the text. 3. How the style and organization of the paper communicates and represents key findings The article is well structured and the main conclusions are well presented. 4. Literature analysis or discussion All the main results have been well discussed in …
Comments to Author
1. Methodological rigour, reproducibility and availability of underlying data The methodology is clearly detailed. The authors provide all the information necessary to ensure the reproducibility of the analyses. 2. Presentation of results The results are well presented. The positives obtained were confirmed by Illumina technology. The authors made a phylogenetic tree from the entire VP1 gene, two complete genomes of EV-G were obtained, the divergence of EV-Gs is well demonstrated in figure 1. These results are robust and clearly presented in the text. 3. How the style and organization of the paper communicates and represents key findings The article is well structured and the main conclusions are well presented. 4. Literature analysis or discussion All the main results have been well discussed in the text. 5. Any other relevant comments Not all of the 27 EV-G sequences obtained in this study appear in the two trees in Figure 2.
Please rate the manuscript for methodological rigour
Very good
Please rate the quality of the presentation and structure of the manuscript
Good
To what extent are the conclusions supported by the data?
Strongly support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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Comments to Author
The Authors tested faecal specimens, collected from pigs and goats in Cameroon and Central African Republic, to understand more about the genetic diversity of enteroviruses in the livestock of these countries. The work is well performed but the Authors are requested to consider the following points. Line 76. The Authors state the EV-Gs were genetically close to EVs described on other continents, but did they mean "other countries"? The three references quoted refer to Central African Republic, Gabon and Nigeria, which are all within the African continent. This also applies to lines 210 and 249. Line 192. The Authors state the EV-Gs were assigned to 11 virus types already described worldwide. However, the Authors only note reports from Cameroon and CAR. Can the Authors provide references with examples …
Comments to Author
The Authors tested faecal specimens, collected from pigs and goats in Cameroon and Central African Republic, to understand more about the genetic diversity of enteroviruses in the livestock of these countries. The work is well performed but the Authors are requested to consider the following points. Line 76. The Authors state the EV-Gs were genetically close to EVs described on other continents, but did they mean "other countries"? The three references quoted refer to Central African Republic, Gabon and Nigeria, which are all within the African continent. This also applies to lines 210 and 249. Line 192. The Authors state the EV-Gs were assigned to 11 virus types already described worldwide. However, the Authors only note reports from Cameroon and CAR. Can the Authors provide references with examples of other countries that EV-Gs were detected worldwide? Are reports worldwide or in several countries in Africa? The description of EV-Gs worldwide also applies to line 36 in the Abstract. Lines 209-211. The authors are requested to provide a reference for the statement concerning the close relationship between EV-Gs from Cameroon and CAR, and different continents. As noted previously, have EV-Gs been reported worldwide or just countries in the African continent? Line 228-229. The Authors report one detection of EV-C99 in pigs and state it is the third report in non-human species, after earlier detections in dogs and chimpanzees. Have the Authors compared the phylogeny of the three EV-C99 sequences from non-human sources? If so, did the three sequences cluster within the same clade or exhibit any shared phylogeny worth noting? General suggestions and grammar Line 171. Correct "reel time RT-PCR" to real-time RT-PCR. The Authors may like to consider the following text as alternatives to what is written in the manuscript: Line 177. "Both primer pairs target conserved sequences flanking the capsid-encoding region…" Line 207. "…sequences do not cluster by virus types…" Line 208. "…takes place between EV-Gs either side of the capsid-encoding region…" Line 243. Delete "to" as an extraneous word to state "…and can be explained by the high number…"
Please rate the manuscript for methodological rigour
Very good
Please rate the quality of the presentation and structure of the manuscript
Very good
To what extent are the conclusions supported by the data?
Strongly support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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