Nutritional sex-specificity on bacterial metabolites during mosquito development leads to adult sex-ratio distortion

Nutritional sex-specificity on bacterial metabolites during mosquito development leads to adult sex-ratio distortion | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Nutritional sex-specificity on bacterial metabolites during mosquito development leads to adult sex-ratio distortion Mathilde Gendrin, Ottavia Romoli, Javier Serrato-Salas, Chloé Gapp, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3944073/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 02 Dec, 2024 Read the published version in Communications Biology → Version 1 posted You are reading this latest preprint version Abstract Mosquitoes rely on their microbiota for B vitamin synthesis. We previously found that Aedes aegypti third-instar larvae cleared of their microbiota were impaired in their development, notably due to a lack of folic acid (vitamin B9). In this study, we investigated the effects of other B vitamins on the development of germ-free mosquito larvae. We found that diet supplementation using a cocktail of seven B vitamins did not improve mosquito developmental success, but rather had a significant impact on the sex-ratio of the resulting adults, with an enrichment of female mosquitoes emerging from B vitamin-treated larvae. A transcriptomic analysis of male and female larvae identified some sex-specific regulated genes upon vitamin treatment. When treating germ-free larvae with high doses of individual B vitamins, we detected a specific toxic effect related to biotin (vitamin B7) exposure at high concentrations. We then provided germ-free larvae with varying biotin doses or with varying bacterial counts, and showed that males are sensitive to biotin toxicity at a lower concentration than females, and require less bacteria-derived nutrients than females. These findings shed new light on sex-specific nutritional requirements and toxicity thresholds during the development of insect larvae, which impact the sex ratio of adults. Biological sciences/Developmental biology Biological sciences/Microbiology Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SuppFigure1.pdf Supplemental Figure S1. Development success of mosquitoes exposed to B vitamins at 1x concentrations during larval development. Proportion of mosquitoes completing their development (dark grey), blocked at the larval stage (white), or dying (light grey) when reversibly colonized until the beginning of the third instar and kept in standard germ-free conditions (GF) or supplemented with a 1x (VIT1x) B vitamin solution. Bar charts represent the mean ± SEM of three independent replicates of 48 mosquitoes per condition. Suppfigure2.pdf Supplemental Figure S2. Experimental set-up and quality check of transcriptomic analysis of male and female larvae exposed to B vitamins. (A) Experimental set-up: mosquito larvae reared in gnotobiotic conditions with the auxotrophic E. coli strain were decolonized at the beginning of the third instar (L3) to obtain germ-free larvae. Larvae were either kept in germ-free conditions or treated with 1x or 8x B vitamin solutions. After three to six hours since moulting to the fourth instar (L4), up to 150 larvae per condition were individually collected. Three independent replicates were performed. DNA was extracted from individual larvae for sex assignment using the Nix gene, while RNA was extracted for transcriptome sequencing on male and female larvae. A quality control step on the proportion of Nix reads and of overall alignment rates and a second check of initial Nix PCR results, allowed to select samples for data analysis. (B) Raw counts of read mapping on the Nix gene in the sequenced transcriptomes. Total number of reads mapping on the Nix gene in the transcriptomes of female and male larvae reversibly colonized at the beginning of the third instar and kept in germ-free conditions (GF) or supplemented with a 1x (VIT1x) or 8x (VIT8x) B vitamin solution. Results from the three replicates (A, B, and C) are shown separately. (C-G) Principal Component Analysis (PCA) of transcriptomes passing the quality check and used for the final analysis. (C) PCA of transcriptomes of female (star) and male (circle) larvae kept in germ-free conditions (white) or treated with vitamins (VIT8x, yellow). (D, E) PCA of transcriptomes of female (star) and male (circle) larvae kept in germ-free conditions (D) or treated with vitamins (E). (F, G) PCA of transcriptomes of male (F) and female (G) larvae kept in germ-free conditions (white) or treated with vitamins (yellow). Suppfigure3.pdf Supplemental Figure S3. Genes differentially regulated in male and female larvae exposed to B vitamins. (A) Number of genes up- (red) and downregulated (blue) in male vs female mosquitoes when larvae were kept in germ-free conditions or treated with the 8x B vitamin solution. The numbers in parenthesis represent the number of genes that are shared between our study and the Matthews et al transcriptome (B) Number of genes up- (red) and downregulated (blue) in B vitamin vs germ-free conditions in male and female larvae. (C, D) List of genes up- (red) and down- regulated (blue) in male (C) and female (D) larvae treated with the B vitamin solution. For each gene, Vectorbase ID, log2 fold value, padj, description, and transcript type (coding gene: yellow; noncoding RNA (ncRNA): green; pseudogene: blue) are indicated. The last column shows paralog or ortholog genes with the name of the species and a brief function description. Asterisks (*) indicate description copied from FlyBase. SuppFigure4.pdf SuppFigure5.pdf SuppFigure6.pdf SupplementalTableS1.pdf Supplemental Table S1. Statistical analyses. For each figure, the type of analysis, response variable (dependent variable), predictor (independent variable), random effect, sample size in each replicate, test results, and results of multiple comparisons (if applicable) are shown. Glmm: generalised linear mixed model. Lsmeans: least square means. SupplementalTableS2.pdf Supplemental Table S2. Genes differentially regulated between male and female larvae kept in the germ-free condition. List of genes up- (red) and down-regulated (blue) in germ-free male vs female larvae, in the absence of vitamin supplementation. For each gene, Vectorbase ID, log 2 fold value, padj, description, and transcript type are indicated. The last column indicates whether the gene was also identified in 23 as differentially regulated in male vs female larvae. SupplementalTableS3.pdf Supplemental Table S3. Genes differentially regulated between male and female larvae treated with the 8x vitamin solution. List of genes up- (red) and down-regulated (blue) in germ-free male vs female larvae treated with the 8x B vitamin solution. For each gene, Vectorbase ID, log 2 fold value, padj, description, and transcript type are indicated. The last column indicates whether the gene was also identified in 23 as differentially regulated in male vs female larvae. SupplementalTableS4.pdf Supplemental Table S4. Sequences of Nix and Actin primers used in this study. For each gene, the Vectorbase accession, 5’-3’ sequence, and references are indicated. Supplementalmaterial.docx Cite Share Download PDF Status: Published Journal Publication published 02 Dec, 2024 Read the published version in Communications Biology → Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3944073","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":273818332,"identity":"0a9119a4-d09c-4132-9687-a287ce059e70","order_by":0,"name":"Mathilde Gendrin","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0001-6405-6458","institution":"Institut Pasteur de la Guyane","correspondingAuthor":true,"prefix":"","firstName":"Mathilde","middleName":"","lastName":"Gendrin","suffix":""},{"id":273818333,"identity":"f2d3e409-91ef-4f22-b274-7ef85a0d60cd","order_by":1,"name":"Ottavia Romoli","email":"","orcid":"","institution":"Institut Pasteur de la Guyane","correspondingAuthor":false,"prefix":"","firstName":"Ottavia","middleName":"","lastName":"Romoli","suffix":""},{"id":273818334,"identity":"a3925e3e-b8b9-4f56-a102-fd3f95aad1a3","order_by":2,"name":"Javier Serrato-Salas","email":"","orcid":"","institution":"Institut Pasteur de la Guyane","correspondingAuthor":false,"prefix":"","firstName":"Javier","middleName":"","lastName":"Serrato-Salas","suffix":""},{"id":273818335,"identity":"8d901013-286b-4073-adec-14f23b65b388","order_by":3,"name":"Chloé Gapp","email":"","orcid":"","institution":"Institut Pasteur de la Guyane","correspondingAuthor":false,"prefix":"","firstName":"Chloé","middleName":"","lastName":"Gapp","suffix":""},{"id":273818336,"identity":"9471801e-4941-4654-bc1f-e241fc107944","order_by":4,"name":"Yanouk Epelboin","email":"","orcid":"","institution":"Institut Pasteur de la Guyane","correspondingAuthor":false,"prefix":"","firstName":"Yanouk","middleName":"","lastName":"Epelboin","suffix":""},{"id":273818337,"identity":"d840f254-de12-4daf-9734-7501c941386f","order_by":5,"name":"Pol Figueras Ivern","email":"","orcid":"","institution":"Institut Pasteur de la Guyane","correspondingAuthor":false,"prefix":"","firstName":"Pol","middleName":"Figueras","lastName":"Ivern","suffix":""},{"id":273818338,"identity":"4a41ea56-cf43-4460-be71-419f1a38116f","order_by":6,"name":"Frédéric Barras","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Frédéric","middleName":"","lastName":"Barras","suffix":""}],"badges":[],"createdAt":"2024-02-09 20:15:10","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3944073/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3944073/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s42003-024-07319-7","type":"published","date":"2024-12-02T05:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":70441705,"identity":"f26eade1-b719-4803-a279-121a29a29232","added_by":"auto","created_at":"2024-12-03 08:07:15","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":941764,"visible":true,"origin":"","legend":"","description":"","filename":"RomoliSerratoSalasmanuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1_covered_befcb8b9-b0c8-43c8-b901-639247f883fa.pdf"},{"id":52104035,"identity":"dd445d9b-1c27-43f7-a683-a7ab4016d569","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":84378,"visible":true,"origin":"","legend":"\u003cp\u003eSupplemental Figure S1. Development success of mosquitoes exposed to B vitamins at 1x\u003c/p\u003e\n\u003cp\u003econcentrations during larval development. Proportion of mosquitoes completing their\u003c/p\u003e\n\u003cp\u003edevelopment (dark grey), blocked at the larval stage (white), or dying (light grey) when reversibly\u003c/p\u003e\n\u003cp\u003ecolonized until the beginning of the third instar and kept in standard germ-free conditions (GF) or\u003c/p\u003e\n\u003cp\u003esupplemented with a 1x (VIT1x) B vitamin solution. Bar charts represent the mean ± SEM of three\u003c/p\u003e\n\u003cp\u003eindependent replicates of 48 mosquitoes per condition.\u003c/p\u003e","description":"","filename":"SuppFigure1.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/b83d5e0ebff6baa58c9a7cb3.pdf"},{"id":52104038,"identity":"f7013efe-0430-4459-b3e7-f185723c7aa4","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":120654,"visible":true,"origin":"","legend":"\u003cp\u003eSupplemental Figure S2. Experimental set-up and quality check of transcriptomic analysis of male\u003c/p\u003e\n\u003cp\u003eand female larvae exposed to B vitamins. (A) Experimental set-up: mosquito larvae reared in\u003c/p\u003e\n\u003cp\u003egnotobiotic conditions with the auxotrophic E. coli strain were decolonized at the beginning of the\u003c/p\u003e\n\u003cp\u003ethird instar (L3) to obtain germ-free larvae. Larvae were either kept in germ-free conditions or\u003c/p\u003e\n\u003cp\u003etreated with 1x or 8x B vitamin solutions. After three to six hours since moulting to the fourth instar\u003c/p\u003e\n\u003cp\u003e(L4), up to 150 larvae per condition were individually collected. Three independent replicates were\u003c/p\u003e\n\u003cp\u003eperformed. DNA was extracted from individual larvae for sex assignment using the Nix gene, while\u003c/p\u003e\n\u003cp\u003eRNA was extracted for transcriptome sequencing on male and female larvae. A quality control step\u003c/p\u003e\n\u003cp\u003eon the proportion of Nix reads and of overall alignment rates and a second check of initial Nix PCR\u003c/p\u003e\n\u003cp\u003eresults, allowed to select samples for data analysis. (B) Raw counts of read mapping on the Nix gene\u003c/p\u003e\n\u003cp\u003ein the sequenced transcriptomes. Total number of reads mapping on the Nix gene in the\u003c/p\u003e\n\u003cp\u003etranscriptomes of female and male larvae reversibly colonized at the beginning of the third instar\u003c/p\u003e\n\u003cp\u003eand kept in germ-free conditions (GF) or supplemented with a 1x (VIT1x) or 8x (VIT8x) B vitamin\u003c/p\u003e\n\u003cp\u003esolution. Results from the three replicates (A, B, and C) are shown separately. (C-G) Principal\u003c/p\u003e\n\u003cp\u003eComponent Analysis (PCA) of transcriptomes passing the quality check and used for the final\u003c/p\u003e\n\u003cp\u003eanalysis. (C) PCA of transcriptomes of female (star) and male (circle) larvae kept in germ-free\u003c/p\u003e\n\u003cp\u003econditions (white) or treated with vitamins (VIT8x, yellow). (D, E) PCA of transcriptomes of female\u003c/p\u003e\n\u003cp\u003e(star) and male (circle) larvae kept in germ-free conditions (D) or treated with vitamins (E). (F, G)\u003c/p\u003e\n\u003cp\u003ePCA of transcriptomes of male (F) and female (G) larvae kept in germ-free conditions (white) or\u003c/p\u003e\n\u003cp\u003etreated with vitamins (yellow).\u003c/p\u003e","description":"","filename":"Suppfigure2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/18ee34ff8630a82585a35c4d.pdf"},{"id":52104042,"identity":"1c89e19c-082f-4f98-bf91-2bf893446844","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":108359,"visible":true,"origin":"","legend":"\u003cp\u003eSupplemental Figure S3. Genes differentially regulated in male and female larvae exposed to B\u003c/p\u003e\n\u003cp\u003evitamins. (A) Number of genes up- (red) and downregulated (blue) in male vs female mosquitoes when larvae were kept in germ-free conditions or treated with the 8x B vitamin solution. The\u003c/p\u003e\n\u003cp\u003enumbers in parenthesis represent the number of genes that are shared between our study and the\u003c/p\u003e\n\u003cp\u003eMatthews et al transcriptome (B) Number of genes up- (red) and downregulated (blue) in B\u003c/p\u003e\n\u003cp\u003evitamin vs germ-free conditions in male and female larvae. (C, D) List of genes up- (red) and down-\u003c/p\u003e\n\u003cp\u003eregulated (blue) in male (C) and female (D) larvae treated with the B vitamin solution. For each gene,\u003c/p\u003e\n\u003cp\u003eVectorbase ID, log2 fold value, padj, description, and transcript type (coding gene: yellow; noncoding\u003c/p\u003e\n\u003cp\u003eRNA (ncRNA): green; pseudogene: blue) are indicated. The last column shows paralog or ortholog\u003c/p\u003e\n\u003cp\u003egenes with the name of the species and a brief function description. Asterisks (*) indicate\u003c/p\u003e\n\u003cp\u003edescription copied from FlyBase.\u003c/p\u003e","description":"","filename":"Suppfigure3.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/ed89f661fa69644321abb3fa.pdf"},{"id":52104037,"identity":"ff229288-80fd-4376-b2c2-b7deb18b1e26","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":129358,"visible":true,"origin":"","legend":"","description":"","filename":"SuppFigure4.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/4c4a30cea7d8c072d9f92e00.pdf"},{"id":52104036,"identity":"be93c214-079e-4af1-88ea-03376f1016b7","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":169100,"visible":true,"origin":"","legend":"","description":"","filename":"SuppFigure5.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/7df6c0675e11633b37214416.pdf"},{"id":52104039,"identity":"cca47b49-cb9f-4325-b843-38a8fb64ea55","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":146901,"visible":true,"origin":"","legend":"","description":"","filename":"SuppFigure6.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/63f8af09ad4b6da870add18f.pdf"},{"id":52104045,"identity":"207146bd-33b6-4065-bd22-593fcd0b6b11","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":7,"title":"","display":"","copyAsset":false,"role":"supplement","size":138555,"visible":true,"origin":"","legend":"\u003cp\u003eSupplemental Table S1. Statistical analyses. For each figure, the type of analysis, response variable\u003c/p\u003e\n\u003cp\u003e(dependent variable), predictor (independent variable), random effect, sample size in each\u003c/p\u003e\n\u003cp\u003ereplicate, test results, and results of multiple comparisons (if applicable) are shown. Glmm:\u003c/p\u003e\n\u003cp\u003egeneralised linear mixed model. Lsmeans: least square means.\u003c/p\u003e","description":"","filename":"SupplementalTableS1.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/6020efdde5bc94c7c686ac9d.pdf"},{"id":52104040,"identity":"9865d3f1-2d26-4984-85d3-b97cab76538b","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":60892,"visible":true,"origin":"","legend":"\u003cp\u003eSupplemental Table S2. Genes differentially regulated between male and female larvae kept in\u003c/p\u003e\n\u003cp\u003ethe germ-free condition. List of genes up- (red) and down-regulated (blue) in germ-free male vs\u003c/p\u003e\n\u003cp\u003efemale larvae, in the absence of vitamin supplementation. For each gene, Vectorbase ID, log\u003csub\u003e2\u003c/sub\u003e fold value, padj, description, and transcript type are indicated. The last column indicates whether the\u003c/p\u003e\n\u003cp\u003egene was also identified in\u003csup\u003e 23 \u003c/sup\u003eas differentially regulated in male vs female larvae.\u003c/p\u003e","description":"","filename":"SupplementalTableS2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/d90a9d6f2f2c565d4b034829.pdf"},{"id":52104044,"identity":"297515ae-c918-4e91-9001-5b43789cae1f","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":9,"title":"","display":"","copyAsset":false,"role":"supplement","size":50655,"visible":true,"origin":"","legend":"\u003cp\u003eSupplemental Table S3. Genes differentially regulated between male and female larvae treated\u003c/p\u003e\n\u003cp\u003ewith the 8x vitamin solution. List of genes up- (red) and down-regulated (blue) in germ-free male\u003c/p\u003e\n\u003cp\u003evs female larvae treated with the 8x B vitamin solution. For each gene, Vectorbase ID, log\u003csub\u003e2\u003c/sub\u003e fold value,\u003c/p\u003e\n\u003cp\u003epadj, description, and transcript type are indicated. The last column indicates whether the gene was\u003c/p\u003e\n\u003cp\u003ealso identified in \u003csup\u003e23\u003c/sup\u003e as differentially regulated in male vs female larvae.\u003c/p\u003e","description":"","filename":"SupplementalTableS3.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/6e0019c04e8819ddf6994828.pdf"},{"id":52104043,"identity":"ba6876e7-26ed-49b6-9f84-42e1f6423bf8","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"pdf","order_by":10,"title":"","display":"","copyAsset":false,"role":"supplement","size":32356,"visible":true,"origin":"","legend":"\u003cp\u003eSupplemental Table S4. Sequences of Nix and Actin primers used in this study. For each gene, the\u003c/p\u003e\n\u003cp\u003eVectorbase accession, 5’-3’ sequence, and references are indicated.\u003c/p\u003e","description":"","filename":"SupplementalTableS4.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/958a9aa160ad9bb8099020c7.pdf"},{"id":52104041,"identity":"0fc46736-3321-42ac-bc9f-20af95fc3cbb","added_by":"auto","created_at":"2024-03-06 19:22:57","extension":"docx","order_by":11,"title":"","display":"","copyAsset":false,"role":"supplement","size":58649,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementalmaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-3944073/v1/230366f4d80104196aaaba8d.docx"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Nutritional sex-specificity on bacterial metabolites during mosquito development leads to adult sex-ratio distortion","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-3944073/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3944073/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Mosquitoes rely on their microbiota for B vitamin synthesis. We previously found that Aedes aegypti third-instar larvae cleared of their microbiota were impaired in their development, notably due to a lack of folic acid (vitamin B9). In this study, we investigated the effects of other B vitamins on the development of germ-free mosquito larvae. We found that diet supplementation using a cocktail of seven B vitamins did not improve mosquito developmental success, but rather had a significant impact on the sex-ratio of the resulting adults, with an enrichment of female mosquitoes emerging from B vitamin-treated larvae. A transcriptomic analysis of male and female larvae identified some sex-specific regulated genes upon vitamin treatment. When treating germ-free larvae with high doses of individual B vitamins, we detected a specific toxic effect related to biotin (vitamin B7) exposure at high concentrations. We then provided germ-free larvae with varying biotin doses or with varying bacterial counts, and showed that males are sensitive to biotin toxicity at a lower concentration than females, and require less bacteria-derived nutrients than females. These findings shed new light on sex-specific nutritional requirements and toxicity thresholds during the development of insect larvae, which impact the sex ratio of adults.","manuscriptTitle":"Nutritional sex-specificity on bacterial metabolites during mosquito development leads to adult sex-ratio distortion","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-06 19:22:52","doi":"10.21203/rs.3.rs-3944073/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"communications-biology","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"commsbio","sideBox":"Learn more about [Communications Biology](http://www.nature.com/commsbio/)","snPcode":"","submissionUrl":"","title":"Communications Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Communications Series","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"34e986dd-e630-4168-9aab-5c6e93f80c4d","owner":[],"postedDate":"March 6th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":28856305,"name":"Biological sciences/Developmental biology"},{"id":28856306,"name":"Biological sciences/Microbiology"}],"tags":[],"updatedAt":"2024-12-03T08:07:07+00:00","versionOfRecord":{"articleIdentity":"rs-3944073","link":"https://doi.org/10.1038/s42003-024-07319-7","journal":{"identity":"communications-biology","isVorOnly":false,"title":"Communications Biology"},"publishedOn":"2024-12-02 05:00:00","publishedOnDateReadable":"December 2nd, 2024"},"versionCreatedAt":"2024-03-06 19:22:52","video":"","vorDoi":"10.1038/s42003-024-07319-7","vorDoiUrl":"https://doi.org/10.1038/s42003-024-07319-7","workflowStages":[]},"version":"v1","identity":"rs-3944073","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3944073","identity":"rs-3944073","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}