{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,13]],"date-time":"2026-03-13T14:25:21Z","timestamp":1773411921632,"version":"3.50.1"},"reference-count":27,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2020,3,10]],"date-time":"2020-03-10T00:00:00Z","timestamp":1583798400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100004423","name":"World Health Organization","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100004423","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000865","name":"Bill and Melinda Gates Foundation","doi-asserted-by":"publisher","award":["OPP1120972"],"award-info":[{"award-number":["OPP1120972"]}],"id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["gatesopenresearch.org"],"crossmark-restriction":false},"short-container-title":["Gates Open Res"],"abstract":"<ns4:p>To eliminate soil-transmitted helminth (STH) infections as a public health problem, the administration of benzimidazole (BZ) drugs to children has recently intensified. But, as drug pressure increases, the development of anthelmintic drug resistance (AR) becomes a major concern. Currently, there is no global surveillance system to monitor drug efficacy and the emergence of AR. Consequently, it is unclear what the current efficacy of the used drugs is and whether AR is already present. The aim of this study is to pilot a global surveillance system to assess anthelmintic drug efficacy and the emergence of AR in STH control programs. For this, we will incorporate drug efficacy trials into national STH control programs of eight countries (Bangladesh, Cambodia, Lao PDR, Vietnam, Ghana, Rwanda, Senegal and a yet to be defined country in the Americas). In each country, one trial will be performed in one program implementation unit to assess the efficacy of BZ drugs against STHs in school-aged children by faecal egg count reduction test. Stool samples will be collected before and after treatment with BZs for Kato-Katz analysis and preserved to purify parasite DNA. The presence and frequency of known single nucleotide polymorphisms (SNPs) in the \u03b2-tubulin genes of the different STHs will subsequently be assessed. This study will provide a global pattern of drug efficacy and emergence of AR in STH control programs. The results will provide complementary insights on the validity of known SNPs in the \u00df-tubulin gene as a marker for AR in human STHs as well as information on the technical and financial resources required to set up a surveillance system. Finally, the collected stool samples will be an important resource to validate different molecular technologies for the detection of AR markers or to identify novel potential molecular markers associated with AR in STH.<\/ns4:p>","DOI":"10.12688\/gatesopenres.13115.1","type":"journal-article","created":{"date-parts":[[2020,3,10]],"date-time":"2020-03-10T08:15:09Z","timestamp":1583828109000},"page":"28","update-policy":"https:\/\/doi.org\/10.12688\/gatesopenres.crossmark-policy","source":"Crossref","is-referenced-by-count":21,"title":["Piloting a surveillance system to monitor the global patterns of drug efficacy and the emergence of anthelmintic resistance in soil-transmitted helminth control programs: a Starworms study protocol"],"prefix":"10.12688","volume":"4","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3601-9825","authenticated-orcid":false,"given":"Johnny","family":"Vlaminck","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Piet","family":"Cools","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marco","family":"Albonico","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shaali","family":"Ame","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thipphavanh","family":"Chanthapaseuth","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vanisaveth","family":"Viengxay","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dung","family":"Do 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Report of a WHO Expert Committee","year":"2002"},{"key":"ref-26","doi-asserted-by":"publisher","first-page":"71-78","DOI":"10.1016\/j.vetpar.2018.02.004","article-title":"How to improve the standardization and the diagnostic performance of the fecal egg count reduction test?","volume":"253","author":"B Levecke","year":"2018","journal-title":"Vet Parasitol."},{"key":"ref-27","article-title":"StarwormsWP02","author":"J Vlaminck","year":"2020"}],"container-title":["Gates Open Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/gatesopenresearch.org\/articles\/4-28\/v1\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/gatesopenresearch.org\/articles\/4-28\/v1\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/gatesopenresearch.org\/articles\/4-28\/v1\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,5,27]],"date-time":"2021-05-27T07:40:33Z","timestamp":1622101233000},"score":1,"resource":{"primary":{"URL":"https:\/\/gatesopenresearch.org\/articles\/4-28\/v1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,10]]},"references-count":27,"URL":"https:\/\/doi.org\/10.12688\/gatesopenres.13115.1","relation":{"has-review":[{"id-type":"doi","id":"10.21956\/gatesopenres.14292.r28674","asserted-by":"subject"},{"id-type":"doi","id":"10.21956\/gatesopenres.14292.r28673","asserted-by":"subject"},{"id-type":"doi","id":"10.21956\/gatesopenres.14292.r28674","asserted-by":"object"},{"id-type":"doi","id":"10.21956\/gatesopenres.14292.r28673","asserted-by":"object"}]},"ISSN":["2572-4754"],"issn-type":[{"value":"2572-4754","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,3,10]]},"assertion":[{"value":"Indexed","URL":"https:\/\/gatesopenresearch.org\/articles\/4-28\/v1#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.21956\/gatesopenres.14292.r28674, D. Timothy J. Littlewood, Department of Life Sciences, Natural History Museum, London, UK, 30 Mar 2020, version 1, indexed","URL":"https:\/\/gatesopenresearch.org\/articles\/4-28\/v1#referee-response-28674","order":0,"name":"referee-response-28674","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Johnny Vlaminck<\/b>; \n<i>Posted: 14 Apr 2020<\/i>; \n<b>First of all we would like to thank Mr. Lttlewood for the time and effort he put into reviewing our manucript and for the questions. Below we have provided a reply for each specific question asked. We hope this can help further clarify the manuscript.  1) <\/b>\n<b>Although the protocol for fixing and storing stool samples for downstream molecular analysis such as qPCR is reasonably well established I would be concerned that a final concentration of 50-70%ethanol (1:1 [stool:100%EtOH] held at room temperature might not be ideal for proposed\/envisaged NGS analyses.<\/b> &nbsp; The protocol for fixing and storing stool samples for downstream molecular analysis has been previously validated by Ayana et al., 2019 in PLoS NTD (DIO: https:\/\/doi.org\/10.1371\/journal.pntd.0007778). To this end, we stored stool aliquots in either 96% ethanol, 5% potassium dichromate or RNA later at 4\u02daC for over a year (425 days). The results of this study highlight that there was little to no reduction in STH specific DNA concentration measured by qPCR. Given the cost associated with RNA-later and both the toxicity and difficulty to acquire potassium-dichromate, we recommended ethanol as the best preservative for stool samples for further molecular research on STH. Our findings were further underpinned by the findings of Papaiakovou et al. 2018 (doi: 10.1371\/ journal.pntd.0006130), who also concluded that, when balancing logistical factors and the need to preserve target DNA, under most circumstances 95% ethanol provides the most pragmatic choice for preserving stool samples in the field. The impact of our storage protocols on NGS has not been explored. Preliminary results of NGS analysis of previously collected and preserved samples (Vlaminck et al., 2019; https:\/\/doi.org\/10.1371\/journal.pntd.0007471) highlight that analyses are not always successful. Failure of analysis mainly observed when the concentration of STH-specific DNA is low. We strongly believe that the amount of STH material included for DNA extraction, and not necessarily the preservation protocol, is the most important parameter for assay success. We are currently exploring different means to increase the DNA yield (extracting more stool \n<i>vs.<\/i> purifying eggs) &nbsp; &nbsp; \n<b>2) I am concerned that resistance detected with a single marker gene will not in itself be efficacious, even before the proposed detection has been established as reliable. Perhaps an understanding that samples (stool\/DNA) will be available for further screening studies within and beyond this study might add value to this wider investment?<\/b> &nbsp; We are aware that the current evidence for the currently known SNPs as a marker for resistance in human STHs is not yet crystal clear. As highlighted in the last paragraph of the discussion of the manuscript, the collected and stored stool samples will allow validation of future molecular technologies to detect drug resistance or to identify novel potential molecular markers associated with resistance in STHs. The samples will be stored at the Neglected Tropical Disease biobank hosted at Ghent University, and they will be incorporated into the virtual STH sample inventory, so that they become available for the wider STH-community. &nbsp; \n<b>3) Is the longevity of the VSSI (and samples) limited by the time-span of the investment or will this somehow be sustained beyond?<\/b> &nbsp; The longevity of the VSSI is not limited to the time-span of the investment. As World Health Organization Collaborating Centre for monitoring the efficacy of anthelminthic drug efficacy for STH (https:\/\/apps.who.int\/whocc\/Detail.aspx?cc_ref=BEL-42&amp;cc_code=bel&amp;), we are putting efforts in place to keep the project website as well as its content online and openly accessible, even after when the current investment ends. &nbsp; \n<b>4) Similarly, publishing data on the project website suggests access will not be sustained; can these data be deposited on a recognised data portal or data site for future access?<\/b> &nbsp; These data will indeed be published on the website. However, the data will also be added as supplementary information to the research papers that describe trial results. To ensure that the wider STH-community has access to our work, we only publish our work in journals that embrace open access, as illustrated by the Starworms collection in PLoS NTD https:\/\/collections.plos.org\/starworms). All the data from our completed studies are available through the supplementary files.","URL":"https:\/\/gatesopenresearch.org\/articles\/4-28\/v1#referee-comment-3309","order":1,"name":"referee-comment-3309","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.21956\/gatesopenres.14292.r28673, Taniawati Supali, Department of Parasitology, Faculty of Medicine, University of Indonesia, Jakarta, Indonesia, 02 Apr 2020, version 1, indexed","URL":"https:\/\/gatesopenresearch.org\/articles\/4-28\/v1#referee-response-28673","order":2,"name":"referee-response-28673","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Johnny Vlaminck<\/b>; \n<i>Posted: 14 Apr 2020<\/i>; \n<b>First of all we would like to thank Mrs. Supali for the time and effort she put into reviewing our manucript and for providing us with some questions. Below we reply to each specific question asked. We hope this further clarifies the manuscript. <\/b> &nbsp; \n<b>1) In the background, the authors wrote that the study would be conducted in eight countries with high Preventive chemotherapy (PC) coverage for at least five years. It is necessary to explain the definition of \u201chigh\u201d in PC coverage \u2013 I would say high is such a relative word, that needs more explanation regarding the number cut offs.&nbsp;In Table 2, the authors wrote the median national coverage of SAC was 59% in Ghana and 60% in Senegal. Are those two countries considered as high PC coverage?&nbsp;If Ghana and Senegal were included as countries with high PC coverage, then I assumed the \u201chigh\u201d range of PC coverage (&gt; 50% - 100% (wide range) was used in the study.<\/b>  Study sites were selected in two consecutive steps. First, countries were selected, after which potential study sites within each country were identified. An important criterion for the selection of the countries was the median national PC coverage of at least 50% (2012 \u2013 2016). Although this value is both arbitrary and low, it allowed the inclusion of countries (i) where PC coverage was increasing over time and (ii) where PC coverage was high in some areas, but low in other areas, this included both Senegal and Ghana. Despite the sometimes low national coverage, the coverage in the selected implementation units in these countries is high (&gt;90%). &nbsp; \n<b>2) In this protocol, the authors were supposed to assess the drug efficacy of BZ drugs (Albendazole and mebendazole).&nbsp; Why were there only two countries (Nicaragua and Cambodia) receiving mebendazole alone?<\/b> &nbsp; The type of benzimidazole drug was not included as a parameter for the country selection. We focused on the coverage of PC program only. As consequence of this, it was purely coincidence that we have two countries receiving mebendazole alone in our final selection. Nevertheless, we foresee that each of the 4 Asian countries (Bangladesh, Cambodia, Lao PDR and Vietnam) will evaluate mebendazole efficacy, while the African countries (Senegal, Ghana and Rwanda) will evaluate albendazole efficacy, and hence we will have efficacy data on both drugs. &nbsp; \n<b>3) There was also a note below Table 2 that Nicaragua has not yet confirmed for the involvement of this study.<\/b> &nbsp; This is correct. At the point of submission Nicaragua has not yet officially confirmed its participation to this study. Co-authors Ana Lucian\u0303ez and Ruben Santiago Nicholls from PAHO are currently following-up on this &nbsp; \n<b>4) It was known as well that the efficacy of Mebendazole is different (see table 4) in Hookworms compared to Albendazole.<\/b> &nbsp; Efficacy of both drugs are indeed different for hookworm infections. This is apparent from the expected efficacy for both drugs (Table 4). Regardless of this difference in intrinsic therapeutic efficacy, it remains crucial to verify any change in efficacy for both drugs in countries where they have been administered on a large scale for a long period of time. &nbsp; \n<b>5) In the methodology, ethanol was needed to preserve the stool for detecting BZ resistance-associated molecular markers. Most of STH endemic areas are remote areas. It would be difficult to bring the ethanol to endemic areas by plane. Several studies had reported that FTA cards can be used to collect the stool samples for PCR. In my opinion, this method would be easier to perform in the field as well as to transport the samples from the endemic areas to the central lab.<\/b> &nbsp; We do not foresee any issues with using ethanol as a preservative in any of our study sites. Discussions with the local research groups has revealed that ethanol is usually easy to obtain. It is also not expected that we will need to take any domestic flights to reach any of the selected study sites. The use of FTA cards could indeed be an alternative and we should look into this for future purposes. However, at this time, we have little experience with this method and our downstream protocol for DNA extraction and sample processing has already been fine-tuned on the use of ethanol-stored stool samples (see also our response to comments made by the other reviewer).","URL":"https:\/\/gatesopenresearch.org\/articles\/4-28\/v1#referee-comment-3310","order":3,"name":"referee-comment-3310","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This work was supported by the Bill and Melinda Gates Foundation [OPP1120972]; and the World Health Organization. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":4,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}