PONE-D-21-33189STUDY ON BOVINE SCHISTOSOMIASIS IN SOME SELECTED AREAS OF SOUTH WOLLO AND OROMIA ZONES OF AMHARA REGION, NORTH-EAST ETHIOPIAPLOS ONE

Dear Dr. Tarekegn Tintagu Gizaw,

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PLOS ONE

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Additional Editor Comments:

In addition the comments of the two reviewers please address the following while revising the manuscript:

1. Please merge study design and study population under materials and methods.

2. Data analysis: Use of Chi-square test only is not enough for the identification of risk factors. The animals are clustered within herd and districts. In this type of data, the mixed-effects logistic regression analysis should be preferred over chi-square test. Initially, the authors should run univariable mixed effect logistic regression model considering district as random intercept [univariable screening]. Then variables with a p-value <0.25 in the univariable screening should be included in the multivariable mixed-effect logistic regression analysis. The authors should describe the methods of model selection, model fit, collinearity, confounding and interaction.

3. The authors tested 384 samples by using both coprological and postmortem methods. So, the authors can easily evaluate the performance [sensitivity, specificity, positive predictive value and negative predictive value] of coprological method considering postmortem as a gold standard. The will add some new results which have diagnostic importance.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Partly

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: No

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: In this study, the authors highlight Schistosoma bovis in cattle relating to its the prevalence and to characterize pathological lesions induced by Schistosoma bovis including risk factor assessment for the animal-level infestation of South Wollo and Oromia Administrative Zones, Amhara regional state, Ethiopia via a cross sectional survey. The study shows the occurrence of S. bovis infection with the overall prevalence of 16.7 % and 17.2 % using fecal sample examination and postmortem examination, respectively. The infection rate was statistically significantly associated with the body condition score, breeds and origins of the studied animals, but not with the sex and age of sampled animal. The overall outlook of the study is interesting. However, there are some lacunae in this manuscript that are needed to address before publication of the manuscript. Details included in the attached file (comments).

Reviewer #2: There are few published articles from the sites included in this study. However, this provides a through characterization of S. bovis from multiple sites. Although the paper contains useful information, I have several concerns that should be addressed before the paper is ready for publication.

General:

Please carefully check the scientific names and make them italic.

I strongly recommend to improve the laboratory coprological examination section with brief description of the laboratory technique.

Please provide necessary references and explanations against the Assumptions made to describe the findings.

Abstract:

1. “In addition to the coprological examination postmortem examination was carried out at Kombolcha and Dessie municipal abattoirs on 384 cattle which came from various livestock markets from the selected districts to recover the adult worms from mesenteric veins and to characterize the gross lesions in liver and intestinal tracts”. Sentence is too long and difficult to understand and realize the meaning. Please rewrite the sentence in break it down in two three sentences.

2. Of the total of 768 faecal samples examined 128 (16.7 %) were found positive for Schistosoma bovis eggs. Provide the 95% CI for the positive percentage.

3. Findings of the abattoir survey showed that 55(14.3%) cattle were positive for schistosoma parasite egg and 66 (17.2%) cattle were found to be positive for adult worm in their mesenteric veins, these findings showed that there was strong measure of agreement (Kappa=0.775) between coprological and post-mortem examination. Provide the 95% CI for the positive percentage.

Introduction:

1. Ethiopia has the largest number of livestock in Africa, [1-2]. Livestock contribute to over 40% of the value of annual agricultural production and no less 15% the gross national product (GNP). Provide few information including total cattle population in the study sites.

2. However, the livestock sector is not efficiently and fully exploited due to several constraints like malnutrition, traditional husbandry practice, poor genetic makeup and prevailing diseases [3-4]. Add few more information about availability or limitations of the veterinary services to diagnose and treat the animal diseases in Ethiopia.

3. In the introduction, write a paragraph on different clinical signs found at different stage of Schistosomiasis in cattle.

4. Therefore, considering the economic importance of the disease, agro ecological variation, different animal husbandry practices and disease prevention and control strategy of the five districts, the current study was designed to determine the prevalence of bovine schistosomiasis (S. bovis), to identify potential risk factors for the occurrence bovine schistosomiasis and to characterize pathological lesions induced by Schistosoma bovis infection. The study objectives is not specific. Please rewrite the study objectives including the study sites name.

Materials and Methods:

1. The study was conducted in some selected districts of South Wollo Zone (Kalu, Tehuledere and Ambasel Districts) around Ardibo, Logo and Golbo lakes and Borkena river) and Oromia Zone at Cheffa valley. Those study areas were selected purposively based the availabilities of permanent water bodies. The word ‘some” is vague. Specify the no. of districts included in the study.

2. Fig 1: From the study map, Amhara district is looking abnormally bigger than others. Please check it and make it clearer to the readers.

3. The study animals were cattle in the field and also the cattle that presented for slaughtering in Dessie and Kombolcha municipal abattoirs from purposely selected study areas without discrimination of their sex, age, origin, breed and body condition. It is quite difficult to understand the district name of Dessie and Kombolcha municipal abattoirs. Rewrite the sentence in which they are situated.

4. Study Design: A cross sectional study was conducted from November 2020 to June 2021 to determine the prevalence of bovine schistosomiasis and associated risk factors. “determine the prevalence of bovine schistosomiasis and associated risk factors” is part of objective. So remove this portion and rewrite the sentence.

5. The cattle population in each study district was obtained from local government veterinary records and the proportion of cattle from each district were weighted according to the estimated population size in that particular district. Create a table and enlist the district names with cattle population from corresponding district.

6. Both in the field and abattoirs the study animals were restrained properly and approximately 10 grams of fresh faecal sample was collected directly from the rectum using gloved hands and then the collected fecal samples were transported with cold chain to Wollo University, School of Veterinary Medicine, Parasitology laboratory. Write this sentences under a separate subheading “sample collection”.

7. What kinds of coprological methods author’s used for this paper is not clear. Please include the methods like direct microscopy or floatation or sedimentation etc. that you have used.

8. The test agreement between Coprological and post-mortem examination was analysed using Kappa statistics. Please mention the different kinds of agreements like weak, moderate, good and strong agreement with references.

9. An official letter of cooperation was obtained from Wollo University ethical clearance committee. Include the issuing no. of the official letter.

Results:

1. Provide 95% CI for all the prevalence or percentage values of S. bovis mentioned throughout the manuscript.

2. Association of Sex, Age, Breed and Body condition of the Study Animals with Schistosoma bovis infection

a. Revise the title as “Association of potential risk factors and Schistosoma bovis infection in cattles of south wollo and oromia zones of amhara region, north-east Ethiopia

b. For this section, firstly write the results with statistical significant values (p<0.005) and then write insignificant findings.

3. Figure 4: Use indicator to identify the specific lesions in the figures as you did for figure 3.

Discussion:

1. The probable reasons for the higher prevalence in the present study might be due to the presence of known water bodies (lakes) in the selected sites may favor the development and multiplication of intermediate hosts and practicing of free grazing on the pasture land might be another predisposing factor. As the authors has no grazing related information of their studied cattles , hence, cite a paper where there is evidence in favor of authors arguments on grazing influencing the prevalence of S. bovis.

2. This is also supported by the explanations of Jesus et al. [40] and Narcis et al. [41] that the prevalence and occurrence of schistosomiasis in a given area could be influenced by local climate conditions, presence of water reservoirs, lakes, rivers, and availability of suitable final and intermediate hosts. In earlier paragraph, you argued that the higher prevalence might be due to the presence of known water bodies (lakes) in the selected sites may favor the development and multiplication of intermediate hosts and now you are giving explanation against that which is not possible. Please read few more papers related to your findings and rewrite the explanation with references.

3. The association of potential host and environmental factors with the occurrence of S.bovis in the selected study sites were assessed and identified. I did not find any environmental factors that you have explored through this study. Please check it.

4. In the present research work prevalence of schistosoma bovis was statistically significantly higher (P<0.005) in local breed animals than the cross breed animals. Write schistosoma bovis as S. bovis and make it italic.

5. This might be due to the fact that local breed cattle were kept in outdoor and repeatedly exposed for Schistosoma infection than the cross breed animals, because cross breeds are kept for the purpose of milking in semi-intensive or intensive management systems with the provision of quality concentrate and roughages feed as well as clean water. Please add references supporting your opinion.

Conclusion and recommendations:

1. Without enlisting the recommendations write them in a single paragraph.

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Reviewer #1: Yes: Sk Shaheenur Islam, Department of Livestock Services, Bangladesh.

Reviewer #2: No

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Attachments

Attachment

Submitted filename: Manuscript_reviewed_track change.doc

Attachment

Submitted filename: Comments_PONE-D-21-33189.docx

PONE-D-21-33189R1Bovine Schistosomiasis in some Selected Areas of South Wollo and Oromia zones of Amhara Region, North-East EthiopiaPLOS ONE

Dear Dr. Gizaw,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

 Academic Editor's comments:Lines 23-27:Please rephrase as suggested below:An abattoir survey was carried out on 384 cattle to evaluate the performance of the sedimentation method. The risk factors were identified using multivariable mixed-effect logistic regression analyses.Please report here the overall prevalence of coprological examination results which also includes the abattoir part. Then report the true prevalence of schistosomiasis based on postmortem results.Please delete  lines 31-41: starting from  "The prevalence" up to "body condition animals"Please rewrite this part by interpreting the odds ratio of three significant variables [breed, body condition, and age] in the multivariable model.Please conclude based on the risk factors. Like, local cattle with medium and poor body condition should be prioritized for deworming and future surveillance.Please delete lines 233-235 up to the citation of R: As univariable mixed-effect logistic regression analyses were performed to assess the association between the outcome and explanatory variables, the Chi-square test is not needed.Line 237 and elsewhere in the manuscript: Please replace univariate with univariable and multivariate with multivariable.Please cut lines 243-250 and paste them at the beginning of the data management and analysis.Please delete lines 273-274: Also, the likelihood ratio (LR) was determined for evaluating the validity of the sedimentation technique using the epiR package.Please add the likelihood ratio in the previous sentence and cite 48 as all of these parameters were estimated using a function of this package. Please delete lines 274 [starting from the "The test] to 278. It is no longer needed when the performance of the sedimentation technique was evaluated which is more informative just than just testing the agreement. Please delete lines 289-295. Please add here the results of the univariable screening. Just mention the name of variables included in the multivariable model citing Table 1. Please merge Tables 1 and 2. It can be done by deleting the last two columns of Table 1 and replacing them with the last two columns of Table 2.The odds ratio estimate shown in Table 2 for sex and age are incorrect.For example, the prevalence of schistosomiasis in male cattle is 15.9%. The male category was considered as a reference [1]. However, the prevalence of schistosomiasis in female cattle is 18.8% higher than that of male cattle. So, the odds ratio for female cattle must be >1 but the authors presented <1. This is also the same for age. In addition, it is better to consider the "young" category of age as a reference for having the lowest prevalence.Please delete this sentence from lines 320-321. They have a significant effect (p<0.05) on bovine Schistosoma.Please delete this sentence from lines 324-325:  Furthermore, young cattle had lower odds (OR=0.64, 95% CI, 0.30-1.35) than old cattle to be shistosoma positive. Because the estimation of the odds ratio for the age group was not correct and also the mentioned odds ratio was not significant. Please recalculate and consider young age as a reference.Please rename the title of Table 5: Evaluation of the efficacy of sedimentation technique for the diagnosis of bovine schistosomiasis (postmortem examination as reference test)Please delete this sentence and Table 4: The findings obtained from the current work showed that there was substantial agreement (Kappa = 0.775) between coprological (sedimentation technique) and post-mortem examination.Please delete lines 431-435 as gender was not significantly associated.Lines 436-445: Here authors made a self-contradictory discussion. Table 3 shows age as a significantly associated variable [although the mentioned odds ratio is not correct] but they are discussing it as non-significant.Please rephrase the long sentence in lines 446-448.Please rephrase the sentence in lines 449-452 [starting from "The current"]Please delete the lines 453-459 as these are results. Please discuss here the six false-positive coprological examination results.lines 467-475:Please rewrite the conclusion based on the results as also suggested earlier.

Please submit your revised manuscript by Jun 09 2022 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

We look forward to receiving your revised manuscript.

Kind regards,

A. K. M. Anisur Rahman, Ph.D.

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

********** 

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The manuscript substantially improved. However, typo-editing errors (scientific name, odd ratio, p value where appropriate) present throughout the text and tables. Author(s) should fix this issue before publication. Additionally, authors should check the plagiarism issue in the final version.

Reviewer #2: Thanks to the authors, they have addressed all the inquiries. Now the manuscript is looking better. I have no further inquiries.

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

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PONE-D-21-33189R2Bovine Schistosomiasis in some Selected Areas of South Wollo and Oromia zones of Amhara Region, North-East EthiopiaPLOS ONE

Dear Mr. Gizaw,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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ACADEMIC EDITOR:Please rephrase Lines 35-38 as given below:Local cattle breed (OR= 2.44, 95%CI= 1.34-4.43), poor body condition (OR= 4.09, 95% CI=2.45-6.83) and adult (OR=1.78, 95% CI= 1.21-3.28) cattle are more likely to acquire schistosomiasis than crossbreed, good body condition, and young cattle.Similarly, please rephrase lines 41-42   In conclusion, adult local cattle with medium and poor body conditions should be prioritized for deworming and future surveillance.Lines 58-63: Please rephrase as, In addition, the existing veterinary service delivery under the current animal health system is considered to be unsatisfactory in both private and public sectors because of the low government attention, inadequate budget allocation, gaps in control of illegal drug circulation, implementation of ethical practices, and shortage of basic laboratory facilities [5].Please rephrase lines 77-81: Out of 10 species reported as naturally infesting cattle, six have received particular attention mainly because of their recognized veterinary significance: please mention those six with veterinary significance [7].Please delete lines: 267-270: "However, the sex of the cattle doesn’t have a significant association (P>0.05) with S. bovis. Accordingly, local breed cattle (OR=2.60; 95% CI= 1.45– 4.67) with poor body condition (OR= 3.70; 95% CI= 1.25–4.18) have a higher risk of acquiring the disease than crossbreed with good body condition cattle." Please start the discussion with the major findings and their importance to control bovine schistosomiasis.Line 363L Please delete "statistically"Please delete "rate" from lines 371, 372, and 376 as prevalence is a proportion [not a rate}.False positivity is related to specificity, not sensitivity. So, if no proper explanation exists then better to delete this sentence: "In this study, six (6) false positive coprological examination results were which might be to the sensitivity (74.24%) of the coprological test relative to postmortem examination."Please delete lines 400-401 as they are general statements, not conclusions: "This study aims to determine the prevalence and associated risk factors of S. bovis in cattle.  In addition, the pathological lesions caused by S. bovis were characterized. Accordingly,"Also, please delete these sentences as they are results, not conclusions: The disease is significantly associated with the age, breed, and body condition of the animals. As a result, adult local cattle with poor body conditions animals were at higher risk to acquire the disease.Please rephrase as, For effective prevention and control of the disease, adult local cattle with medium and poor body conditions should be prioritized for deworming and future surveillance.=====================

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A. K. M. Anisur Rahman, Ph.D.

Academic Editor

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Bovine Schistosomiasis in some Selected Areas of South Wollo and Oromia zones of Amhara Region, North-East Ethiopia

PONE-D-21-33189R3

Dear Dr. Gizaw,

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A. K. M. Anisur Rahman, Ph.D.

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