Environmental sampling for the detection of capripox viruses and peste des petits ruminants virus in households and livestock markets in Plateau State, Nigeria

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Abstract

Multiple transboundary animal diseases (TADs) circulate in Plateau State, Nigeria, where livestock keeping is common and contributes to both the physical and socio-economic well-being of a large proportion of the population. In this study, we explored the potential for environmental sampling to detect viruses causing TADs circulating in the region. Electrostatic dust cloths were used to swab areas of the environment likely to have contact with secretions and excretions from infected animals. Samples were collected monthly from five households, one transhumance site and one livestock market in two local government areas in Plateau State between March and October 2021. These were tested for the presence of peste des petits ruminants virus (PPRV) and capripox viruses using real-time PCR. Of the 458 samples collected, 2.4% ( n = 11) were positive for PPRV RNA and 1.3 % ( n = 6) were positive for capripox virus DNA. A capripox differentiation assay showed that these samples were positive for sheep pox virus ( n = 2), goat pox virus ( n = 2) and lumpy skin disease virus ( n = 2). Our results demonstrate that environmental sampling could be used as part of TAD surveillance in the area. Environmental swabs require little technical knowledge to collect and can be used to detect multiple viruses from a single sample.

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  1. Dear Dr. Simon Gubbins, Thank you very much for submitting your manuscript "Environmental sampling for the detection of capripox viruses and peste des petits ruminants virus in households and livestock markets in Northern Nigeria" for consideration at Access Microbiology. Your manuscript was reviewed by members of the editorial board and by two independent reviewers. In light of the reviews, we are pleased to inform you that the manuscript is provisionally accepted for publication. We believe the study would be a valuable contribution to the existing literature. The reviewers have highlighted minor concerns. Please ensure that you address their comments. When you are ready to resubmit, please upload a detailed list of your responses to the reviewer comments and a description of the changes you have made in the manuscript. Your revised manuscript is also likely to be sent to reviewers for re-evaluation. Please prepare and submit your revised manuscript within 30 days. If you anticipate any delay, please let us know. We hope that you find the peer review process constructive, and we welcome your feedback at any time. Thank you again for your submission. Sincerely,

  2. Comments to Author

    In the research methodology of this manuscript, the description is detailed and well-supported by clear references. The results are presented both in the form of figures and tables, which are appropriately organized and compiled into the article, enhancing the clarity of the content for readers. However, there are some minor amendments that need to be addressed, including additional data presentation and discussion, to further enhance the completeness of the article. Please refer to the recommendations below. Abstract - Line 24: Could you please specify the total number of collected samples and the number of positive samples before calculating and reporting the percentage of positives? - The environmental sampling method used to collect samples in this study should be declared in your abstract. Introduction - Line 49-55: The importance of these diseases should be explained in terms of the morbidity and mortality implications for the ruminant livestock industry, to highlight the significance of this disease surveillance. - Please specify the stability of each pathogen in the environment to justify the use of environmental sampling methods for detecting the pathogens, rather than sampling directly from the animals. - Line 66-67: The introduction should include a detailed comparison of diagnostic methods for the disease, including previous surveillance techniques. Describe how these methods differ in terms of advantages and disadvantages compared to environmental sampling and explain how molecular methods provide advantages over traditional approaches. Materials and Methods - Line 74: What criteria were used to select these three sampling locations, and is there any transportation of animals between them? - Line 80: The time period for sample collection specified in the Materials and Methods does not align with the details provided in the legend of Figure 1. Please review and ensure consistency. - Line 81: A timeline for sample collection should be included, along with details on the types of sampling sites and the number of samples collected. This information should be presented in a summary table. Results - Line 115: The location where GTPV was found in your manuscript does not align with the description in Table 1. Please review and correct this discrepancy. Discussion - Generally, these pathogens can be shed through animal secretions, leading to environmental contamination. Therefore, it is crucial to establish criteria for sampling at various sites to accurately represent pathogen contamination and assess persistence in the environment. Additionally, although the sampling method is non-invasive for the animals, using swabs for environmental sampling may introduce contamination from organic matter, which could potentially affect pathogen detection. Please provide a more detailed discussion on these issues.

    Please rate the manuscript for methodological rigour

    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

  3. Comments to Author

    This study is an offshoot of previous research by the authors that showed that oral and environmental swabs are adequate sampling methods for the early detection of ruminant diseases, especially Foot-and-mouth disease, at animal and herd levels. The objective is to detect etiological agents of transboundary animal diseases (TADs) from environmental samples. The authors collected environmental samples with electrostatic dust cloths from households, transhuman sites, and livestock markets in two Local Government Areas (LGAs) in Plateau State, Nigeria. Samples were processed and then assessed for peste de petit ruminants virus (PPRV) RNA and capripoxvirus DNA via PCR. A species-specific capripoxvirus differentiation assay specifically detected sheeppox virus (SPPV), goatpox virus (GTPV), and lumpy skin disease virus (LSDV). The authors showed data that suggest environmental samples are suitable for TAD surveillance and detection. Nevertheless, there are a couple of suggestions that can make the manuscript better. The authors mentioned that TADs negatively affect animal production, food supply, and production costs, but they did not present any specific figures or data to highlight this impact, especially in the study location. The authors can add morbidity and mortality figures of each TAD mentioned to underscore the socioeconomic importance. The positivity rate of detection from the environmental samples collected is low. Many samples must be collected before achieving single-digit positivity rates based on the data presented. The authors can address this in the discussion section to justify the proposal of environmental sampling despite the seemingly non-commensurate balance between efforts and results. This study happened in two Local Government Areas (LGAs) of Plateau State, one of the nineteen Northern States in Nigeria. Data from this single state is unlikely to be representative of all nineteen. Thus, the authors may consider adjusting the title to reflect that this study occurred in a state located in Northern Nigeria. 1. Methodological rigour, reproducibility and availability of underlying data This was a non-complex study that involved collection of environmental samples using electrostatic dust clothes and ultimate detection of the nucleic acids of some viruses using PCR. The PCR methods were also referenced. The methodology should be reproducible and underlying data was provided. 2. Presentation of results The major results were provided in Table 1 and Supplementary Data 1. The results were presented adequately in the manuscript. 3. How the style and organization of the paper communicates and represents key findings The key findings were well-communicated in the paper and adequately captured in Figure 1. 4. Literature analysis or discussion The utility of environmental samples for detection of TADs was adequately discussed. However, the authors can also give mortality and morbidity figures to highlight the socioeconomic importance of the viruses detected. 5. Any other relevant comments There may be a need to readjust the title of the paper. This study was carried out in Plateau State, one of the nineteen Northen states in Nigeria. The data from only one Northern State cannot be representative of all nineteen. The percentage of positivity from the environmental samples was quite low. Do the authors have any reason(s) or discussion regarding this? This can be added to the discussion section of the paper.

    Please rate the manuscript for methodological rigour

    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