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Harpur Fellowship in Parasitology, McGill University"},{"name":"Graduate Excellence Fellowship, McGill University"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Parasites Vectors"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Current measures for the prevention of dirofilariasis, caused by the dog heartworm,\n                      <jats:italic>Dirofilaria immitis<\/jats:italic>\n                      , rely on macrocyclic lactones, but evidence of drug-resistant isolates has called for alternative approaches to disease intervention. As microfilariae are known to be in a state of developmental arrest in their mammalian host and then undergo two molts once inside the arthropod, the aim of this study was to look at the developmental regulation of\n                      <jats:italic>D. immitis<\/jats:italic>\n                      microfilariae that occurs in their arthropod host using\n                      <jats:italic>in vitro<\/jats:italic>\n                      approaches and to investigate the role of the ecdysone signaling system in this development regulation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Dirofilaria immitis<\/jats:italic>\n                      microfilariae extracted from dog blood were incubated under various culture conditions to identify those most suitable for\n                      <jats:italic>in vitro<\/jats:italic>\n                      culture and development of the microfilariae, and to determine the effects of fetal bovine serum (FBS), mosquito cells, and ecdysteroid on the development of the microfilariae. Transcript levels of the ecdysone signaling pathway components were measured with droplet digital PCR (ddPCR).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      <jats:italic>In vitro<\/jats:italic>\n                      conditions that best promote early development of\n                      <jats:italic>D. immitis<\/jats:italic>\n                      microfilariae to the \u201clate sausage stage\u201d have been identified, although shedding of the cuticle was not observed. FBS had inhibitory effects on the development and motility of the microfilariae, but media conditioned with\n                      <jats:italic>Anopheles gambiae<\/jats:italic>\n                      cells were favorable to microfilarial growth. The transcript level study using ddPCR also showed that ecdysone signaling system components were upregulated in developing microfilariae and that 20-hydroxyecdysone increased the proportion of larvae developing to the sausage and late sausage stages\n                      <jats:italic>in vitro<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      The arthropod host environment provides cues required for the rapid development of\n                      <jats:italic>D. immitis<\/jats:italic>\n                      microfilariae, and the ecdysone signaling system may play an important role in filarial nematode developmental transitions. This study contributes to a better understanding of the developmental process of\n                      <jats:italic>D. immitis<\/jats:italic>\n                      microfilariae.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical Abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1186\/s13071-020-04480-w","type":"journal-article","created":{"date-parts":[[2020,12,9]],"date-time":"2020-12-09T06:23:52Z","timestamp":1607495032000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Developmental regulation of Dirofilaria immitis microfilariae and evaluation of ecdysone signaling pathway transcript level using droplet digital PCR"],"prefix":"10.1186","volume":"13","author":[{"given":"Tsai-Chi","family":"Shang Kuan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roger K.","family":"Prichard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,12,9]]},"reference":[{"key":"4480_CR1","unstructured":"American Veterinary Medical Association. https:\/\/www.avma.org\/. 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