{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T05:10:28Z","timestamp":1772169028158,"version":"3.50.1"},"update-to":[{"DOI":"10.12688\/wellcomeopenres.14555.1","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2018,8,15]],"date-time":"2018-08-15T00:00:00Z","timestamp":1534291200000}}],"reference-count":52,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2018,8,15]],"date-time":"2018-08-15T00:00:00Z","timestamp":1534291200000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Sciences Research Council","doi-asserted-by":"crossref","award":["BB\/H022406\/1"],"award-info":[{"award-number":["BB\/H022406\/1"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Wellcome Trust","award":["101285"],"award-info":[{"award-number":["101285"]}]}],"content-domain":{"domain":["wellcomeopenresearch.org"],"crossmark-restriction":false},"short-container-title":["Wellcome Open Res"],"abstract":"<ns5:p>\n                    Leishmaniases is a group of vector-borne parasitic diseases transmitted by sand flies that affects 1.3 million people across 98 countries, with limited control strategies due to the lack of an available vaccine and the emergence of insecticide resistance.\u00a0 Novel control strategies that are being explored for mosquito-borne diseases, such as\u00a0\n                    <ns5:italic>Wolbachia\u00a0<\/ns5:italic>\n                    bacterial inhibition of pathogens and genetically modified insects (e.g. using CRISPR-Cas9 editing), rely on the ability to consistently inject eggs of the target species.\u00a0 Here we present a novel method to obtain and inject preblastoderm sand fly eggs of the genus\u00a0\n                    <ns5:italic>Lutzomyia (Lu.)\u00a0<\/ns5:italic>\n                    <ns5:italic>longipalpis<\/ns5:italic>\n                    , the principle vector of zoonotic visceral leishmaniasis in South America. The procedures required to obtain sufficiently young\u00a0\n                    <ns5:italic>Lu. longipalpis\u00a0<\/ns5:italic>\n                    colony eggs are described alongside a microinjection technique that permits rapid injection and minimal handling of small sand fly eggs post-injection. Using a strain of\u00a0\n                    <ns5:italic>Wolbachia\u00a0<\/ns5:italic>\n                    as a \u2018marker\u2019 for successful injection, our protocol produced early generation\u00a0\n                    <ns5:italic>Wolbachia\u00a0<\/ns5:italic>\n                    transinfected\u00a0\n                    <ns5:italic>Lu. longipalpis\u00a0<\/ns5:italic>\n                    lines, demonstrating its potential as the first step for use in novel applied strategies for sand fly control.\n                  <\/ns5:p>","DOI":"10.12688\/wellcomeopenres.14555.2","type":"journal-article","created":{"date-parts":[[2018,8,15]],"date-time":"2018-08-15T07:35:08Z","timestamp":1534318508000},"page":"55","update-policy":"https:\/\/doi.org\/10.12688\/wellcomeopenres.crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment of a method for Lutzomyia longipalpis sand fly egg microinjection: The first step towards potential novel control strategies for leishmaniasis"],"prefix":"10.12688","volume":"3","author":[{"given":"Claire L.","family":"Jeffries","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew E.","family":"Rogers","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3545-012X","authenticated-orcid":false,"given":"Thomas","family":"Walker","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2560","published-online":{"date-parts":[[2018,8,15]]},"reference":[{"key":"ref-1","doi-asserted-by":"publisher","first-page":"873-882","DOI":"10.1038\/nrmicro1748","article-title":"Visceral leishmaniasis: what are the needs for diagnosis, treatment and control?","volume":"5","author":"F Chappuis","year":"2007","journal-title":"Nat Rev 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A male-determining factor in the mosquito Aedes aegypti.","volume":"348","author":"A Hall","year":"2015","journal-title":"Science."},{"key":"ref-51","doi-asserted-by":"publisher","first-page":"78-83","DOI":"10.1038\/nbt.3439","article-title":"A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles gambiae.","volume":"34","author":"A Hammond","year":"2016","journal-title":"Nat Biotechnol."},{"key":"ref-52","article-title":"Establishment of a Method for Lutzomyia Longipalpis Sand Fly Embryo Microinjection: The First Step towards Potential Novel Control Strategies for Leishmaniasis.","author":"T Walker","year":"2018","journal-title":"Open Science Framework."}],"container-title":["Wellcome Open Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/wellcomeopenresearch.org\/articles\/3-55\/v2\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,5,19]],"date-time":"2020-05-19T20:10:05Z","timestamp":1589919005000},"score":1,"resource":{"primary":{"URL":"https:\/\/wellcomeopenresearch.org\/articles\/3-55\/v2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,8,15]]},"references-count":52,"URL":"https:\/\/doi.org\/10.12688\/wellcomeopenres.14555.2","relation":{"has-review":[{"id-type":"doi","id":"10.21956\/wellcomeopenres.16029.r33697","asserted-by":"subject"},{"id-type":"doi","id":"10.21956\/wellcomeopenres.16029.r33697","asserted-by":"object"}]},"ISSN":["2398-502X"],"issn-type":[{"value":"2398-502X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,8,15]]},"assertion":[{"value":"Indexed","URL":"https:\/\/wellcomeopenresearch.org\/articles\/3-55\/v2#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.21956\/wellcomeopenres.15842.r33300, Fernando Ariel Genta, Laboratory of Insect Physiology and Biochemistry, Oswaldo Cruz Institute - Oswaldo Cruz Foundation (IOC-FIOCRUZ), Rio de Janeiro, Brazil, 22 Jun 2018, version 1, 1 approved, 1 approved with reservations","URL":"https:\/\/wellcomeopenresearch.org\/articles\/3-55\/v1#referee-response-33300","order":0,"name":"referee-response-33300","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Thomas Walker<\/b>; \n<i>Posted: 07 Aug 2018<\/i>; Many thanks Fernando for your review and we have addressed your comments as follows (in bold):&nbsp;  Abstract line 9 - &quot;principal&quot; instead of &quot;principle&quot; \n<b>We have changed this in the manuscript<\/b>  Results page 6, sentence &quot;These initial tests...&quot; I understand that they are initial tests, but I missed a more detailed description and comparison of the data. How much better was the chosen condition in comparison to the others? How many eggs were laid in the three media tested? How strong was the effect of light and sand fly number? These considerations would be important for developers and to have a better understanding of the sand fly biology involved. \n<b>This was indeed more observational during initial tests to indicate the suitability of using agar as a substrate for oviposition (plaster of paris used for colony maintenance).&nbsp;&nbsp;As we found that sandflies laid a significant proportion of melanised eggs during the initial trials, we went on to determine the proportion of \u2018injectable\u2019 eggs on days 3-5 post-bloodfeed (Figure 3, raw data file) as that would have a greater impact on the ability to undertake this work rather than overall numbers of eggs laid.&nbsp;&nbsp;<\/b>  Figure 3. The legend of the figure does not describe all charts in part b. Maybe it is just a problem in the colors described, they do not match all the pizza slices in the figure. &nbsp;\n<b>We agree and have modified the figure legend to clarify the difference between egg colour from melanisation and colours on the charts!&nbsp;<\/b>&nbsp; &nbsp; Discussion page 9. Sentence &quot;Secondly, the rapid mortality...&quot; We have just published a report showing second blood feeding and oviposition in sand flies (\n<i>Lutzomyia longipalpis<\/i>&nbsp;of the very same strain used in this article) in large-scale&nbsp;experiments with the same efficiency when compared to the first blood meal\/oviposition cycle\n<u><sup>1<\/sup><\/u>. I think our finding might be useful for the authors in the future and it would be interesting to mention them in this context. &nbsp;\n<b>We have now included this result and reference in our discussion.&nbsp;&nbsp; &nbsp;<\/b>","URL":"https:\/\/wellcomeopenresearch.org\/articles\/3-55\/v2#referee-comment-3665","order":1,"name":"referee-comment-3665","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.21956\/wellcomeopenres.16029.r33697, Marcelo Ramalho-Ortigao, F. Edward Hebert School of Medicine, Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences (USUHS), Bethesda, MD, USA, 22 Aug 2018, version 2, indexed","URL":"https:\/\/wellcomeopenresearch.org\/articles\/3-55\/v2#referee-response-33697","order":2,"name":"referee-response-33697","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"CLJ and TW were supported by a Wellcome Trust\/Royal Society Sir Henry Dale fellowship awarded to TW (101285\/Z\/13\/Z). MER was supported by a BBSRC David Phillips Fellowship (BB\/H022406\/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":3,"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"}]}}