PONE-D-24-02504Pathogenic characteristics of an aggregated diarrhea event caused by Pseudomonas shigelliides from streamPLOS ONE

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

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Reviewer #1: Reviewer's Comments on the Manuscript:

The paper studied combines epidemiological investigation with laboratory results, analyzing an aggregation diarrhea event which caused by Pseudomonas shigelliides. Pseudomonas shigelliides as an enteropathogen are often underestimated and receive insufficient attention in the world. Most reports of P. shigelliides outbreaks are more years old. This study isolated P. shigelliides from patients and water samples, then analyzing pathogenic characteristics of the strains by drug sensitivity test, PFGE and WGS. The study has a good indicative effect for food contaminated by P. shigelliides in water. But there are some obstacles for the readers to understand, and the following comments should be considered in a revision.

1. Abstract, the content description is inaccurate. The author should state the time of event clearly, because there may be not only one aggregation diarrhea event in Huzhou,2023. It is suggested to describe it as “trace the origin of P. shigelliides” rather than “trace the origin of the pathogenic bacteria”. It is suggested to delete the "of the cases" in line 12.

2.The English expression in the paper needs to be consistent for clarity. After the first appearance of "Pseudomonas shigella", "P. shigelliides" can be used as a substitute in the following. The same to "WGS" replace " whole genome sequencing".

3. Sample collection and detection, it should be noted that which 22 pathogens can be detected by Filmarray GI test strip.

4. Sample collection and detection, which paper or methods were referenced for the isolation and culture. P. shigelliides does not have standards for isolation now.

5. Sequence analysis, the genome sequences of the 7 P. shigelloides isolates should be submitted to GenBank/DDBJ/ENA and acquired the accession numbers.

6. Discussion, in paragraph 1, it is recommended to add a description of local distribution of P. shigelliides formerly and the measures taken by the CDC after this diarrhea aggregation event.

7. Discussion, antibiotic resistance, especially multidrug resistance(MDR), is becoming increasingly severe. It is suggested to add discussion of MDR in paragraph 3.

8. Discussion, in paragraph 6,this study found the most closely related strain was GCF_959021465.1_DRR221358_bin.47_MetaWRAP_v1.3_MAG_genomic. But there is no description of the differences in the number of SNP variation. It is suggested to supplement the explanation in the paper.

Overall, the research holds promise and contributes meaningfully to the field. It is one of the few reports in recent years regarding the outbreak of P. shigelloides. In addition to patients, strains were also isolated from the stream water that is important for studying of P. shigelloides. However, addressing the aforementioned concerns and making the suggested modifications will undoubtedly strengthen the paper, ensuring its readiness. It is suggested to publish after modification.

Reviewer #2: 1. Please verify, it is ‘Pseudomonas shigelliides’ or ‘Pseudomonas shigelloides’ or ‘Plesiomonas shigelloides’. In all the case it seems not ‘Pseudomonas’ See this article : Janda JM, Abbott SL, McIver CJ. Plesiomonas shigelloides Revisited. Clin Microbiol Rev. 2016;29(2):349-74. REALLY IT IS NOT ALLOWED TO MAKE THIS MISTAKES OF CONFUSION.

2. Please correct ‘Gram-negative’ not ‘gram-negative'. Verify this in all your article (see section discussion also)

3. In ‘diarrheal Escherichia coli’, the term ‘diarrheal’ should not be in italic.

4. In section ‘Sample collection and detection’, is this correct ‘pathogens, including Pseudomonas shigella’? please verify all your manuscript according to my first comment.

5. The term ‘MAC’ means ‘MacKonkey’ ? please write the full term without abbreviation, at least for the first time that you mention it.

6. Change the term’ Drug sensitivity test’ by ‘antimicrobial susceptibility test’

7. In all the manuscript, please verify punctuation and there are a lot of word not separated (especially in the section ‘antimicrobial susceptibility test’. Please never write the name of antibiotics with capital letters, write ‘ampicillin (AMP), ampicillin/sulbactam (AMS), cefotaxime (CTX), ceftazidime (CAZ), cefoxitin (CFX), Cefazolin (CFZ), imipenem (IMP), cefotaxime/clavulanic acid (CTX/C), ceftazidime/clavulanic acid (CAZ/C), amoxicilin/clavulanic acid (AMC), cefuroxime (CXM), cefepime (CPM), ceftazidime/avibactam (CZA), meropenem (MEM), ertapenem(ETP), ceftiofur (CEF), ciprofloxacin (CIP), nalidixic acid (NAL), gentamicin (Gen), amikacin(AMK), streptomycin (STR), tetracycline (TET), tigecycline (TIG), florfenicol (FFC), colistin (CT), Polymixin (PB), azithromycin (AZM), trimethoprim-sulfamethoxazole (SXT), and chloramphenicol (CHL). E. coli ATCC29522 was used as quality control strain’. THE ‘E. coli’ must be in italic.

8. The name of the enzyme used in PFGE must be in italic ‘Spe’ and ‘I’ not in italic, this is also for ‘Xba’

9. In ‘Salmonellaenterica serovar Braenderup’ make space between ‘Salmonella’ and enterica’ and write them in italic.

10. Please in section results, mention that kpnH, rsmA, rsmA, adeF and CRP are ‘antibiotic efflux pumps’ or ‘efflux resistance genes’. This could explain the multidrug resistance phenotype caused by each pump.

11. Please correct as follows:’.. Greenlees et al. [18]conducted a study that revealed a higher incidence of reported cases during the warmer months when water temperatures increase, facilitating the proliferation of plesiomonads through sewage contamination.’

12. Please in discussion, add 3 to 4 sentences describing the antibiotic efflux pumps (kpnH, rsmA, rsmA, adeF, CRP).

13. You said ‘which was submitted in June 2023,’ Please say from which country this strain was collected, and its source (food, water, patient???).

14. In conclusion, add ‘s” (several genes), so write ‘All isolates carry the same drug resistance genes and virulence genes except cgsE , and the homology between isolates is high.’

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Reviewer #1: Yes: Qingli Dong

Reviewer #2: Yes: Mohamed Salah Abbassi

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Submitted filename: PONE-D-24-02504.docm

Pathogenic characteristics of an aggregated diarrhea event caused by Plesiomonas shigelloides from stream

PONE-D-24-02504R1

Dear Dr. Yan,

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Kind regards,

Furqan Kabir

Academic Editor

PLOS ONE

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