Do wild-caught Drosophilids cooperatively forage?

Do wild-caught Drosophilids cooperatively forage? | 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 Research Article Do wild-caught Drosophilids cooperatively forage? Reeves Kuhar, Madeline Williamson, Peyton Yee, Guzel Naik, Sean Michael Cursain, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4669901/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 26 Nov, 2024 Read the published version in Journal of Comparative Physiology A → Version 1 posted 9 You are reading this latest preprint version Abstract Animals often form organized cooperative foraging groups, where individual members must adhere to specific rules to maintain cohesiveness. These groups face the challenge of managing potential intruders, who may or may not assist in foraging. In semi-liquid food environments, Drosophila larvae learn to synchronize their movements into clusters, which are thought to make feeding more efficient. Individuals who do not synchronize with the group are excluded from the cluster. Whether clustering behavior occurs in wild-caught larvae, and if so, the extent of their selectivity in group membership, remains unknown. Here, we show that clustering occurs across a number of Drosophilid species, and the capacity to join different clusters varies both between and within species. We collected and observed a larval cluster from rotting fruit in the field, yielding seven Drosophilid species. Subsequent tests for clustering on five stable lines from this collection and 20 other inbred wild-caught lines revealed that all species, except D. suzukii, exhibit clustering behavior. Each line demonstrates varying capacities to become members of different clusters. Additionally, combinations of wild species with lab benchmark strains give varied outcomes in resultant adult fitness. The ability to co-cluster varies between and within species boundaries. However, fly lines that cluster with another tend to impart fitness both to themselves and their host. Our findings demonstrate that multiple species of fly larvae can co-cluster, with variations in clustering ability likely due to genetic factors. This behavior tends to confer mutual benefits to cluster members, suggesting significant ecological implications in Drosophila communities. Drosophila foraging cooperation social behavior group membership fitness wild type behavior Figures Figure 1 Figure 2 Figure 3 Figure 4 Full Text Additional Declarations No competing interests reported. Supplementary Files figs1a.mp4 Supplementary figures Movie s1A Video of an outside cluster on a tomato. This was collected and the resultant larvae used to make lines for this study. figs1b.mp4 Movie s1B Video of 2D clustering of F1 mixed larvae obtained from the wild tomato cluster. FigS2a.tiff Figure s2A Images of clusters in vials of 25 wild-caught Drosophilid lines of which 24 cluster. FigS2b.mp4 Movie s2B Example video of Megaselia scalaris clustering in 2D. FigS2czap.mp4 Movie s2C Example video of transplanted Zap into CS 2D clusters. FigS2dsim2.mp4 Movie s2D Example video of transplanted sim2 in CS 2D clusters. FigS2emeg.mp4 Movie s2E Example video of transplanted sim2 in CS 2D clusters at high resolution. FigS2fhiressim2.mp4 Movie s2F Example video of transplanted sim2 in CS 2D clusters at high resolution. FigS2ghiresDmR.mp4 Movie s2G Example video of transplanted DmR in CS 2D clusters at high resolution. Cite Share Download PDF Status: Published Journal Publication published 26 Nov, 2024 Read the published version in Journal of Comparative Physiology A → Version 1 posted Editorial decision: Revision requested 14 Aug, 2024 Reviews received at journal 14 Aug, 2024 Reviews received at journal 27 Jul, 2024 Reviewers agreed at journal 19 Jul, 2024 Reviewers agreed at journal 16 Jul, 2024 Reviewers invited by journal 06 Jul, 2024 Editor assigned by journal 03 Jul, 2024 Submission checks completed at journal 03 Jul, 2024 First submitted to journal 01 Jul, 2024 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. 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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-4669901","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":329336347,"identity":"52ae45da-a30a-4253-9b08-7ba12d231326","order_by":0,"name":"Reeves Kuhar","email":"","orcid":"","institution":"University of Virginia","correspondingAuthor":false,"prefix":"","firstName":"Reeves","middleName":"","lastName":"Kuhar","suffix":""},{"id":329336348,"identity":"591ac1ff-ff8e-4f0a-8dab-a673854aca68","order_by":1,"name":"Madeline Williamson","email":"","orcid":"","institution":"University of Virginia","correspondingAuthor":false,"prefix":"","firstName":"Madeline","middleName":"","lastName":"Williamson","suffix":""},{"id":329336350,"identity":"2e41f591-819a-46f9-925c-898b86212732","order_by":2,"name":"Peyton Yee","email":"","orcid":"","institution":"University of Virginia","correspondingAuthor":false,"prefix":"","firstName":"Peyton","middleName":"","lastName":"Yee","suffix":""},{"id":329336351,"identity":"0353a5fd-47fb-4ded-879e-3f0668d2a029","order_by":3,"name":"Guzel Naik","email":"","orcid":"","institution":"University of Virginia","correspondingAuthor":false,"prefix":"","firstName":"Guzel","middleName":"","lastName":"Naik","suffix":""},{"id":329336356,"identity":"84953ccf-108c-4f7a-ae46-cbdb09d5d5bd","order_by":4,"name":"Sean Michael Cursain","email":"","orcid":"","institution":"University of Virginia","correspondingAuthor":false,"prefix":"","firstName":"Sean","middleName":"Michael","lastName":"Cursain","suffix":""},{"id":329336357,"identity":"151bdda8-011e-4f7d-a80a-664928749dc9","order_by":5,"name":"Barry Condron","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1ElEQVRIiWNgGAWjYJACCSCWY2AGkglgNoMBUVqMSdeS2IAkgF8Lf3vvwdsVFXfSNxznffzi4Q4LeQb25m0SeG04cy7Z8syZZ7kbDrObWSSekTBs4DlWhleLgUSOmWRj2+Hcmc1sbAaJbRKMDUARorSkS0K12DfIvyFOSwI/MxvzA6CWxAYJHvxaJM6cMbZsOHPYsJ+ZjY0BqCW5jSet2AKfFv72HsObDRWH5dn4jzF//NlWZ9vPfnjjDXxakAEb2D1sxCoHAeYPpKgeBaNgFIyCkQMA6NhBie7mYYsAAAAASUVORK5CYII=","orcid":"","institution":"University of Virginia","correspondingAuthor":true,"prefix":"","firstName":"Barry","middleName":"","lastName":"Condron","suffix":""}],"badges":[],"createdAt":"2024-07-01 18:53:31","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4669901/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4669901/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00359-024-01724-3","type":"published","date":"2024-11-26T15:58:33+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":61193946,"identity":"0bfc4b8b-95e8-40b1-a2ef-7b1627318bd5","added_by":"auto","created_at":"2024-07-26 21:07:15","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":12428964,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eObtaining clustering larvae from the wild\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e1A.\u003c/strong\u003e A tomato that had fallen from a vine had ruptured and the center liquified with visible small larvae. The tomato was cut (dashed white line) and a 50x75mm glass slide was placed against the liquified center. Cluster-like larval groups were seen almost immediately. These were video recorded.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e1B.\u003c/strong\u003e Two still photos from a video shows what fulfill the definition of a cluster(Dombrovski et al. 2017). A 2 cm cube of material including the cluster in B was removed and placed in a vial.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e1C.\u003c/strong\u003e Clusters from the tomato sample were seen in the vial. These larvae were incubated to adults and 7 species of flies and one small wasp hatched. These were allowed to reproduce and lay eggs in a separate vial. From this F1 vial, only 5 species hatched.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e1D.\u003c/strong\u003e Samples from this mixed species F1 vial were tested for clustering in 2D assays using CS processed food. Adults hatching from this vial were separated by species into breeding pairs.\u003c/p\u003e","description":"","filename":"fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/420095f6eb5ed9c7be574b46.png"},{"id":61193947,"identity":"d017a24d-a4e2-4bd7-ab9d-81e634333767","added_by":"auto","created_at":"2024-07-26 21:07:15","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":6479615,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eclustering parameters vary for wild caught larvae\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2A.\u003c/strong\u003e Each species was grouped in separate vials and produced 5 lines of 5 species. From each line, single sibs were chosen and crossed. This was done \u0026gt;10 times to make inbred lines. These were combined with 19 previously established inbred lines (see materials) for further study. All together, these wild flies originate from 4 locations in the Eastern US. The relationships of the 5 species are indicated and the ones used in this study are shaded in blue. Calliphoraida, which might also cluster(Scanvion et al. 2018), was not used in this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2B.\u003c/strong\u003e At least 50 adults from each of the 24 lines were placed in egg cups and 24hr old larvae were obtained. About 200 of these larvae were placed in CS-processed vials and clustering was monitored over a few days. Clusters were observed in all vials except that of D. suzukii. See Fig s2A. 9 lines of 5 species were chosen for further study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2C.\u003c/strong\u003e Larvae were allowed to cluster in CS-processed vials as described(Dombrovski et al. 2017). 40 larvae were removed and placed in a 2D apparatus with CS-processed food. The percentage of larvae in clusters at 4,5 and 6 hours was averaged. All larvae cluster with the exception of D. suzukii. Almost all larvae of Megaselia larvae are in clusters. Points indicate average values and error bars represent the standard error. The number of 2D samples used is indicated on the Xaxis. Statistical probabilities were calculated by ANOVA, after normality tests, followed by Tukey’s method. The probabilities compared to CS are indicated with significance marked by P\u0026lt;0.01 ** and P\u0026lt;0.001 ***. A T-test was performed between simR and sim2 and is significant P\u0026lt;0.0001.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2D.\u003c/strong\u003e Clustering residing times of transplanted larvae. Third instar larvae were removed from crowded vials with clusters and labeled with food coloring. Single larvae were placed over clusters of host larvae and the residence time was measured as described(Dombrovski et al. 2017, 2019). With the exception of M1 and suz, all spend 20-60’ in clusters. suz larvae do not enter CS clusters. There is also asymmetry in many transplantations, like M1-\u0026gt;CS/CS-\u0026gt;M1, DmR-\u0026gt;CS/CS-\u0026gt;DmR and sim2-\u0026gt;CS/CS-\u0026gt;sim2. Points indicate average values and error bars represent the standard error. The number of 2D samples used is indicated on the X-axis. Statistical probabilities were calculated by ANOVA, after normality test, followed by Tukey’s method. The probabilities compared to CS are indicated with significance marked by P\u0026lt;0.01 **, P\u0026lt;0.001 *** and P\u0026lt;0.0001 ****. A T-test was performed between the 3 reciprocal pairs.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2E. \u003c/strong\u003eInter-larval synchrony time was measured for transplanted larvae and compared to residing time in clusters. When measured for the different wild types, synchrony explains some of the variance in cluster residence time. However, some cases of long residence time, like sim2-\u0026gt;CS, seems to happen without a high degree of synchrony.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2F. \u003c/strong\u003eTo test for the potential effects of species-specific food processing, food from 3 species was used to host CS 2D clusters. Plotted is the proportion of CS larvae clustering and resultant wing size of emerging adults from matched vials. Compared to food processed by CS, that processed by megaselia, zaprionus and simulans supports CS is clustering and gives healthy adult wing sizes. Only zaprious food gives lower wing sizes for CS. The data points are the averages of the indicated number of samples. The error bars indicate standard deviation for wing size and standard error for clustering. Statistics were calculated by normality tests followed by ANOVA and Tukey’s test. The probabilities are indicated and * is P\u0026lt;0.05.\u003c/p\u003e","description":"","filename":"fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/7985f11877ae25234338e698.png"},{"id":61193944,"identity":"e3b4cdc9-2ba5-4d6f-84a7-3dff91dcbf07","added_by":"auto","created_at":"2024-07-26 21:07:15","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":5110972,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e3A-3I: \u003c/strong\u003eLarva fitness was measured by emerging female adult wing size from growth in a vial. Second instar larvae were placed in a vial, as 10x, 40x, 10x and 30x wild type D.melanogaster host ‘P, and 30x with 10xP. The effects of intra-specific clustering can be seen in any change from 10x to 40x. The effects of heterospecific blends can be measured by comparing 40x with either 10-30 or 30-10 blends with P. The effects on P are summarized in the right graph for each panel. The blue line represents the average 40xP wing size. The individual wing sizes are shown with the red line marking the mean. Statistical significance was measured by ANOVA post a normality test followed by Tukey’s method. The upper of the two probabilities compares to 40x and the lower to 10x of the same kind of larvae. P\u0026lt;0.05 *, P\u0026lt;0.01 **, P\u0026lt;0.001 *** and P\u0026lt;0.0001 ****. For P host measurements, the probability is compared to 40xP(Liao et al. 2024b).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3A CS:\u003c/strong\u003e data is replotted and analyzed from before(Liao et al. 2024b). CS gains more from P than P from CS\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3B\u003c/strong\u003e DmR neither gains nor loses to P.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3C\u003c/strong\u003e M1 loses fitness to P and P remains the same.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3D\u003c/strong\u003e C8 and P lose fitness in blends.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3E\u003c/strong\u003e simR and P lose fitness in some blends.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3F\u003c/strong\u003e sim2 loses in self clustering, and also loses in blends while P gains.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3G\u003c/strong\u003e suz loses in blends while P gains.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3H\u003c/strong\u003e zap loses in blends while P remains neutral.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3I\u003c/strong\u003e meg and P lose fitness in some blends.\u003c/p\u003e\n\u003cp\u003e3J Summary of fitness gains or losses in blends with P for each of the 9 lines tested. An arrow pointing to the P host indicates a gain in fitness of P and an arrow to the test line a gain of that line. The self referential arrows indicate gains of self clusters. The large arrows indicate gains while the small arrows either a loss or neutral effect. There is not a species pattern to the gains or losses.\u003c/p\u003e","description":"","filename":"fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/3066d55116ff4ad72016a18c.png"},{"id":61193945,"identity":"4fe3b0ef-806b-4f9b-b00c-6ee2e294bc15","added_by":"auto","created_at":"2024-07-26 21:07:15","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":2250406,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCorrelations of larval blends\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eClustering and fitness data from wild caught flies were assembled and correlations examined. Four selected lines are indicated for each graph.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4A\u003c/strong\u003e The relationship between the amount of clustering any line conducted with itself was compared to the gain from that behavior. The gain from self clustering was measured by wing size for 40x larvae in a vial divided by that from only 10 larvae. This was compared to proportion of larvae that cluster in 2D.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4B \u003c/strong\u003eThe gain in fitness of an intruder in P was measured by wing size at 10/30 blend compared to 40 alone. This is plotted against time spent by an intruder in a 2D P cluster.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4C\u003c/strong\u003e The same relationship in 4B is now plotted but now as the gain of the host versus the time spent by the intruder in that host cluster.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4D \u003c/strong\u003eGain of intruder is plotted against the gain of the host.\u003c/p\u003e","description":"","filename":"fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/99f614a54b0a0e47a48fd3b3.png"},{"id":70391289,"identity":"e3fb80eb-3ba1-43ec-a793-781779ab26aa","added_by":"auto","created_at":"2024-12-02 17:30:36","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":8140795,"visible":true,"origin":"","legend":"","description":"","filename":"wtpaperspring2024v5.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1_covered_67d599d5-9c74-44c3-910b-422ce7e40fad.pdf"},{"id":61193949,"identity":"c37b2796-be77-44e2-bdf3-0d73ee2a9414","added_by":"auto","created_at":"2024-07-26 21:07:16","extension":"mp4","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":27014982,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary figures\u003c/p\u003e\n\u003cp\u003eMovie s1A\u003c/p\u003e\n\u003cp\u003eVideo of an outside cluster on a tomato. This was collected and the resultant larvae used to make lines for this study.\u003c/p\u003e","description":"","filename":"figs1a.mp4","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/5aeec71e1119882879302c73.mp4"},{"id":61193948,"identity":"c69bcac3-b7b6-4987-ad83-12b79cf11f93","added_by":"auto","created_at":"2024-07-26 21:07:16","extension":"mp4","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":17061960,"visible":true,"origin":"","legend":"\u003cp\u003eMovie s1B\u003c/p\u003e\n\u003cp\u003eVideo of 2D clustering of F1 mixed larvae obtained from the wild tomato cluster.\u003c/p\u003e","description":"","filename":"figs1b.mp4","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/de0bab525883962ccc5368b7.mp4"},{"id":61193954,"identity":"83d3d298-b043-4cd2-8119-1f7beb911e60","added_by":"auto","created_at":"2024-07-26 21:07:30","extension":"tiff","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":296412158,"visible":true,"origin":"","legend":"\u003cp\u003eFigure s2A\u003c/p\u003e\n\u003cp\u003eImages of clusters in vials of 25 wild-caught Drosophilid lines of which 24 cluster.\u003c/p\u003e","description":"","filename":"FigS2a.tiff","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/5132b73a0f3140a1722b0b6f.tiff"},{"id":61193953,"identity":"4ef85410-23ad-41a3-b67a-7e5abbfe23ae","added_by":"auto","created_at":"2024-07-26 21:07:24","extension":"mp4","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":156418772,"visible":true,"origin":"","legend":"\u003cp\u003eMovie s2B\u003c/p\u003e\n\u003cp\u003eExample video of Megaselia scalaris clustering in 2D.\u003c/p\u003e","description":"","filename":"FigS2b.mp4","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/a27af502ed7d80804b056b31.mp4"},{"id":61193951,"identity":"00aa038f-fc77-44c1-8f04-dc93053f0464","added_by":"auto","created_at":"2024-07-26 21:07:22","extension":"mp4","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":132134247,"visible":true,"origin":"","legend":"\u003cp\u003eMovie s2C\u003c/p\u003e\n\u003cp\u003eExample video of transplanted Zap into CS 2D clusters.\u003c/p\u003e","description":"","filename":"FigS2czap.mp4","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/1f4f2a8468bd4391eb778c97.mp4"},{"id":61193952,"identity":"70f8bacf-8f29-4a72-b202-e2deaa4b1442","added_by":"auto","created_at":"2024-07-26 21:07:22","extension":"mp4","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":123704373,"visible":true,"origin":"","legend":"\u003cp\u003eMovie 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21:07:36","extension":"mp4","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":396181619,"visible":true,"origin":"","legend":"\u003cp\u003eMovie s2F\u003c/p\u003e\n\u003cp\u003eExample video of transplanted sim2 in CS 2D clusters at high resolution.\u003c/p\u003e","description":"","filename":"FigS2fhiressim2.mp4","url":"https://assets-eu.researchsquare.com/files/rs-4669901/v1/b59c9c2ad03fe1860c9cdce8.mp4"},{"id":61193974,"identity":"8d462c69-2ccc-4316-8388-3944b22738aa","added_by":"auto","created_at":"2024-07-26 21:08:01","extension":"mp4","order_by":9,"title":"","display":"","copyAsset":false,"role":"supplement","size":895841092,"visible":true,"origin":"","legend":"\u003cp\u003eMovie s2G\u003c/p\u003e\n\u003cp\u003eExample video of transplanted DmR in CS 2D clusters at high 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Physiology A](http://link.springer.com/journal/359)","snPcode":"359","submissionUrl":"https://submission.nature.com/new-submission/359/3","title":"Journal of Comparative Physiology A","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Drosophila, foraging, cooperation, social behavior, group membership, fitness, wild type behavior","lastPublishedDoi":"10.21203/rs.3.rs-4669901/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4669901/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Animals often form organized cooperative foraging groups, where individual members must adhere to specific rules to maintain cohesiveness. These groups face the challenge of managing potential intruders, who may or may not assist in foraging. In semi-liquid food environments, Drosophila larvae learn to synchronize their movements into clusters, which are thought to make feeding more efficient. Individuals who do not synchronize with the group are excluded from the cluster. Whether clustering behavior occurs in wild-caught larvae, and if so, the extent of their selectivity in group membership, remains unknown. Here, we show that clustering occurs across a number of Drosophilid species, and the capacity to join different clusters varies both between and within species. We collected and observed a larval cluster from rotting fruit in the field, yielding seven Drosophilid species. Subsequent tests for clustering on five stable lines from this collection and 20 other inbred wild-caught lines revealed that all species, except D. suzukii, exhibit clustering behavior. Each line demonstrates varying capacities to become members of different clusters. Additionally, combinations of wild species with lab benchmark strains give varied outcomes in resultant adult fitness. The ability to co-cluster varies between and within species boundaries. However, fly lines that cluster with another tend to impart fitness both to themselves and their host. Our findings demonstrate that multiple species of fly larvae can co-cluster, with variations in clustering ability likely due to genetic factors. This behavior tends to confer mutual benefits to cluster members, suggesting significant ecological implications in Drosophila communities. 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