{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T22:30:24Z","timestamp":1768516224738,"version":"3.49.0"},"posted":{"date-parts":[[2024,10,24]]},"group-title":"Molecular Biology","reference-count":115,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,10,24]],"date-time":"2024-10-24T00:00:00Z","timestamp":1729728000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,10,24]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Tsetse flies (\n                  <jats:italic>Glossina<\/jats:italic>\n                  spp.) vector African trypanosomes that cause devastating diseases in humans and domestic animals. Within the\n                  <jats:italic>Glossina<\/jats:italic>\n                  genus, species in the Palpalis subgroup exhibit greater resistance to trypanosome infections compared to those in the\n                  <jats:italic>Morsitans<\/jats:italic>\n                  subgroup. Varying microbiota composition and species-specific genetic traits can significantly influence the efficiency of parasite transmission. Notably, infections with the endosymbiotic bacterium\n                  <jats:italic>Spiroplasma<\/jats:italic>\n                  have been documented in several Palpalis subgroup species, including\n                  <jats:italic>Glossina fuscipes fuscipes<\/jats:italic>\n                  (\n                  <jats:italic>Gff<\/jats:italic>\n                  ). While\n                  <jats:italic>Spiroplasma<\/jats:italic>\n                  infections in\n                  <jats:italic>Gff<\/jats:italic>\n                  are known to hinder trypanosome transmission, the underlying mechanisms remain unknown. To investigate\n                  <jats:italic>Spiroplasma-<\/jats:italic>\n                  mediated factors affecting\n                  <jats:italic>Gff<\/jats:italic>\n                  vector competence, we conducted high-throughput RNA sequencing of the midgut tissue along with functional assays. Our findings reveal elevated oxidative stress in the midgut environment in the presence of\n                  <jats:italic>Spiroplasma<\/jats:italic>\n                  , evidenced by increased expression of\n                  <jats:italic>nitric oxide synthase,<\/jats:italic>\n                  which catalyzes the production of trypanocidal nitric oxide. Additionally, we observed impaired lipid biosynthesis leading to a reduction of this important class of nutrients essential for parasite and host physiologies. In contrast, trypanosome infections in\n                  <jats:italic>Gff\u2019s<\/jats:italic>\n                  midgut significantly upregulated various immunity-related genes, including a small peptide,\n                  <jats:italic>Stomoxyn-like<\/jats:italic>\n                  , homologous to Stomoxyns first discovered in the stable fly\n                  <jats:italic>Stomoxys calcitrans<\/jats:italic>\n                  . We observed that the\n                  <jats:italic>Stomoxyn-like<\/jats:italic>\n                  locus is exclusive to the genomes of\n                  <jats:italic>Palpalis<\/jats:italic>\n                  subgroup tsetse species.\n                  <jats:italic>Gff<\/jats:italic>\n                  Stomoxyn is constitutively expressed in the cardia (proventriculus) and synthetic\n                  <jats:italic>Gff<\/jats:italic>\n                  Stomoxyn exhibits potent activity against\n                  <jats:italic>Escherichia coli<\/jats:italic>\n                  and bloodstream form of\n                  <jats:italic>Trypanosoma brucei<\/jats:italic>\n                  parasites, while showing no effect against insect stage procyclic forms or tsetse\u2019s commensal endosymbiont\n                  <jats:italic>Sodalis in vitro<\/jats:italic>\n                  . Reducing\n                  <jats:italic>Gff<\/jats:italic>\n                  Stomoxyn levels significantly increased trypanosome infection prevalence, indicating its potential trypanocidal role\n                  <jats:italic>in vivo<\/jats:italic>\n                  . Collectively, our results suggest that the enhanced resistance to trypanosomes observed in\n                  <jats:italic>Spiroplasma<\/jats:italic>\n                  -infected\n                  <jats:italic>Gff<\/jats:italic>\n                  may be due to the reduced lipid availability necessary for parasite metabolic maintenance. Furthermore,\n                  <jats:italic>Gff<\/jats:italic>\n                  Stomoxyn could play a crucial role in the initial immune response(s) against mammalian parasites early in the infection process in the midgut and prevent gut colonization. We discuss the molecular characteristics of\n                  <jats:italic>Gff<\/jats:italic>\n                  Stomoxyn, its spatial and temporal expression regulation and its microbicidal activity against\n                  <jats:italic>Trypanosome<\/jats:italic>\n                  parasites. Our findings reinforce the nutritional influences of microbiota on host physiology and host-pathogen dynamics.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Author Summary<\/jats:title>\n                  <jats:p>\n                    The tsetse fly,\n                    <jats:italic>Glossina fuscipes fuscipes<\/jats:italic>\n                    (\n                    <jats:italic>Gff<\/jats:italic>\n                    ) is of high public health relevance. Gff exhibits strong innate resistance to trypanosomes, especially when infected with the endosymbiotic bacterium\n                    <jats:italic>Spiroplasma<\/jats:italic>\n                    . This study investigated how the bacterium\n                    <jats:italic>Spiroplasma<\/jats:italic>\n                    inside\n                    <jats:italic>Gff<\/jats:italic>\n                    enables them to be resistant to trypanosome infection. Our results indicate alterations in host lipid metabolism with reduction in levels of triglycerides, suggesting a potential metabolic barrier that limits the viability to parasite. In addition, we discovered a small peptide, stomoxyn, exclusively in\n                    <jats:italic>Gff<\/jats:italic>\n                    and related\n                    <jats:italic>Palpalis<\/jats:italic>\n                    tsetse species. We have shown that\n                    <jats:italic>Gff<\/jats:italic>\n                    synthetic Stomoxyn has antibacterial and antitrypanosomal properties and lowering Stomoxyn levels in\n                    <jats:italic>Gff<\/jats:italic>\n                    correlates with increased parasite prevalence. We suggest that strategies to increase\n                    <jats:italic>Spiroplasma<\/jats:italic>\n                    prevalence or enhance stomoxyn expression through paratransgenic approaches could be promising avenues for reducing trypanosomiasis transmission.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2024.10.24.620045","type":"posted-content","created":{"date-parts":[[2024,10,25]],"date-time":"2024-10-25T01:45:16Z","timestamp":1729820716000},"source":"Crossref","is-referenced-by-count":1,"title":["<i>Spiroplasma<\/i>\n                  endosymbiont reduction of host lipid synthesis and Stomoxyn-like peptide contribute to trypanosome resistance in the tsetse fly\n                  <i>Glossina fuscipes<\/i>"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1330-8296","authenticated-orcid":false,"given":"Erick","family":"Awuoche","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gretchen","family":"Smallenberger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Bruzzese","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessandra","family":"Orfano","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brian L.","family":"Weiss","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9941-143X","authenticated-orcid":false,"given":"Serap","family":"Aksoy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024102813250491000_2024.10.24.620045v1.1","unstructured":"WHO. 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