{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T18:21:55Z","timestamp":1774635715912,"version":"3.50.1"},"reference-count":55,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T00:00:00Z","timestamp":1774569600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005416","name":"Norges Forskningsr\u00e5d","doi-asserted-by":"publisher","award":["335244"],"award-info":[{"award-number":["335244"]}],"id":[{"id":"10.13039\/501100005416","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    Honeybees (\n                    <jats:italic>Apis mellifera<\/jats:italic>\n                    ) provide crucial pollination services to agricultural systems globally, however, their healthspan in these contexts is constantly at risk. Agricultural environments impose a variety of sublethal stressors onto honeybees, including parasites, pathogens, pesticides, and poor nutrition. Synergies between age, age-associated tasks, and these stressors are believed to underlie colony failure trends of the past decade. Identifying the mechanisms by which age and stressors impact honeybee physiology is an important priority in protecting honeybee and other pollinator populations. An underexplored physiological structure in honeybees is the blood-brain barrier, a protective layer of cells that surrounds the brain. Here, we assess key dimensions of blood-brain barrier function; paracellular and transcellular permeability to molecules in the hemolymph. We measured these modes of permeability across worker groups that differ in age and foraging experience, as well as in bees exposed to varying levels of infestation by the parasitic mite\n                    <jats:italic>Varroa destructor<\/jats:italic>\n                    during development. Our results demonstrate that the paracellular permeability of the honeybee blood-brain barrier is stable across these age groups and upon\n                    <jats:italic>Varroa<\/jats:italic>\n                    exposure. In contrast, we found that transcellular permeability is increased in honeybees exposed to a high\n                    <jats:italic>Varroa<\/jats:italic>\n                    load. Together, these results demonstrate that age-related variation and parasitic stress differentially impact a primary protective structure of the honeybee central nervous system, which may lead to targeted interventions for protecting honeybee healthspan. The assay developed here may be easily applied to different aging- and stress-related contexts, further enabling studies focused on understanding maintenance and decline of the honeybee blood-brain barrier.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0343142","type":"journal-article","created":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T17:45:32Z","timestamp":1774633532000},"page":"e0343142","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Effect of aging and Varroa parasitism on the paracellular and transcellular permeability of the honeybee blood-brain barrier"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7648-246X","authenticated-orcid":true,"given":"Tyler","family":"Quigley","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gro","family":"Amdam","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,3,27]]},"reference":[{"issue":"2","key":"pone.0343142.ref001","doi-asserted-by":"crossref","DOI":"10.1073\/pnas.2002548117","article-title":"Agricultural intensification and climate change are rapidly decreasing insect biodiversity","volume":"118","author":"PH Raven","year":"2021","journal-title":"Proc Natl Acad Sci U S A"},{"key":"pone.0343142.ref002","volume-title":"The assessment report of the intergovernmental science-policy platform on biodiversity and ecosystem services on pollinators, pollination and food production","author":"IPBES","year":"2016"},{"issue":"1608","key":"pone.0343142.ref003","first-page":"303","article-title":"Importance of pollinators in changing landscapes for world crops","volume":"274","author":"A-M Klein","year":"2007","journal-title":"Proc Biol Sci"},{"issue":"8","key":"pone.0343142.ref004","doi-asserted-by":"crossref","first-page":"688","DOI":"10.3390\/insects12080688","article-title":"Overview of bee pollination and its economic value for crop production","volume":"12","author":"SAM Khalifa","year":"2021","journal-title":"Insects"},{"key":"pone.0343142.ref005","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/j.cois.2018.02.004","article-title":"Drivers of colony losses","volume":"26","author":"N Steinhauer","year":"2018","journal-title":"Curr Opin Insect Sci"},{"key":"pone.0343142.ref006","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1146\/annurev-animal-020518-115045","article-title":"Defining pollinator health: a holistic approach based on ecological, genetic, and physiological factors","volume":"8","author":"MM L\u00f3pez-Uribe","year":"2020","journal-title":"Annu Rev Anim Biosci"},{"key":"pone.0343142.ref007","unstructured":"USDA (US Department of Agriculture. Annual strategic pollinator priorities report. 2022."},{"key":"pone.0343142.ref008","unstructured":"US Department of Agriculture USDA. Science and research strategy, 2023-2026. 2023."},{"issue":"1","key":"pone.0343142.ref009","doi-asserted-by":"crossref","first-page":"74","DOI":"10.2174\/1874213000902010074","article-title":"The economic value of the pollination service, a review across scales","volume":"2","author":"L Hein","year":"2009","journal-title":"The Open Ecol J"},{"issue":"1","key":"pone.0343142.ref010","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/00218839.2023.2264601","article-title":"A national survey of managed honey bee colony losses in the USA: results from the Bee informed partnership for 2020\u201321 and 2021\u201322","volume":"63","author":"D Aurell","year":"2023","journal-title":"J Apicult Res"},{"key":"pone.0343142.ref011","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.cois.2015.04.004","article-title":"Death of the bee hive: understanding the failure of an insect society","volume":"10","author":"AB Barron","year":"2015","journal-title":"Curr Opin Insect Sci"},{"issue":"14","key":"pone.0343142.ref012","doi-asserted-by":"crossref","first-page":"8252","DOI":"10.1021\/acs.est.9b02452","article-title":"Traces of a neonicotinoid induce precocious foraging and reduce foraging performance in honey bees","volume":"53","author":"T Colin","year":"2019","journal-title":"Environ Sci Technol"},{"issue":"5","key":"pone.0343142.ref013","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1093\/cz\/zoab002","article-title":"Varroa destructor mite infestations in capped brood cells of honeybee workers affect emergence development and adult foraging ability","volume":"67","author":"H Yang","year":"2021","journal-title":"Curr Zool"},{"issue":"4","key":"pone.0343142.ref014","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.tree.2016.12.009","article-title":"Why bees are so vulnerable to environmental stressors","volume":"32","author":"S Klein","year":"2017","journal-title":"Trends Ecol Evol"},{"issue":"11","key":"pone.0343142.ref015","doi-asserted-by":"crossref","first-page":"758","DOI":"10.1038\/nrn3357","article-title":"The honeybee as a model for understanding the basis of cognition","volume":"13","author":"R Menzel","year":"2012","journal-title":"Nat Rev Neurosci"},{"issue":"1823","key":"pone.0343142.ref016","doi-asserted-by":"crossref","first-page":"20190738","DOI":"10.1098\/rstb.2019.0738","article-title":"Social modulation of ageing: mechanisms, ecology, evolution","volume":"376","author":"TP Quigley","year":"2021","journal-title":"Philos Trans R Soc Lond B Biol Sci"},{"issue":"1","key":"pone.0343142.ref017","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.nbd.2009.07.030","article-title":"Structure and function of the blood-brain barrier","volume":"37","author":"NJ Abbott","year":"2010","journal-title":"Neurobiol Dis"},{"key":"pone.0343142.ref018","doi-asserted-by":"crossref","first-page":"414","DOI":"10.3389\/fnins.2014.00414","article-title":"Barrier mechanisms in the Drosophila blood-brain barrier","volume":"8","author":"SJ Hindle","year":"2014","journal-title":"Front Neurosci"},{"key":"pone.0343142.ref019","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.nbd.2017.02.008","article-title":"Metabolite transport across the mammalian and insect brain diffusion barriers","volume":"107","author":"A Weiler","year":"2017","journal-title":"Neurobiol Dis"},{"key":"pone.0343142.ref020","article-title":"Brain endothelial cells are exquisite sensors of age-related circulatory cues","volume":"30","author":"MB Chen","year":"2020","journal-title":"Cell Reports"},{"key":"pone.0343142.ref021","doi-asserted-by":"crossref","first-page":"26331055221120252","DOI":"10.1177\/26331055221120252","article-title":"The fly blood-brain barrier fights against nutritional stress","volume":"17","author":"EG Contreras","year":"2022","journal-title":"Neurosci Insights"},{"key":"pone.0343142.ref022","doi-asserted-by":"crossref","unstructured":"DeSalvo MK. The Drosophila surface glia transcriptome: evolutionarily conserved blood-brain barrier processes. 2014.","DOI":"10.3389\/fnins.2014.00346"},{"issue":"5","key":"pone.0343142.ref023","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1016\/j.celrep.2017.10.026","article-title":"Evolutionarily conserved roles for blood-brain barrier xenobiotic transporters in endogenous steroid partitioning and behavior","volume":"21","author":"SJ Hindle","year":"2017","journal-title":"Cell Rep"},{"key":"pone.0343142.ref024","doi-asserted-by":"crossref","first-page":"106071","DOI":"10.1016\/j.nbd.2023.106071","article-title":"The Drosophila blood-brain barrier emerges as a model for understanding human brain diseases","volume":"180","author":"EG Contreras","year":"2023","journal-title":"Neurobiol Dis"},{"issue":"3","key":"pone.0343142.ref025","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1111\/nan.12408","article-title":"Age-associated changes in the blood-brain barrier: comparative studies in human and mouse","volume":"44","author":"EF Goodall","year":"2018","journal-title":"Neuropathol Appl Neurobiol"},{"key":"pone.0343142.ref026","doi-asserted-by":"crossref","first-page":"404","DOI":"10.3389\/fnins.2014.00404","article-title":"The rights and wrongs of blood-brain barrier permeability studies: a walk through 100 years of history","volume":"8","author":"NR Saunders","year":"2014","journal-title":"Front Neurosci"},{"issue":"22","key":"pone.0343142.ref027","doi-asserted-by":"crossref","first-page":"12111","DOI":"10.3390\/ijms222212111","article-title":"Form and function of the vertebrate and invertebrate blood-brain barriers","volume":"22","author":"AD Dunton","year":"2021","journal-title":"Int J Mol Sci"},{"issue":"5","key":"pone.0343142.ref028","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/j.tins.2020.12.002","article-title":"Blood-brain barrier dynamics to maintain brain homeostasis","volume":"44","author":"M Segarra","year":"2021","journal-title":"Trends Neurosci"},{"key":"pone.0343142.ref029","doi-asserted-by":"crossref","first-page":"405","DOI":"10.3389\/fncel.2019.00405","article-title":"The evolving concept of the blood brain barrier (BBB): from a single static barrier to a heterogeneous and dynamic relay center","volume":"13","author":"A Villabona-Rueda","year":"2019","journal-title":"Front Cell Neurosci"},{"issue":"1","key":"pone.0343142.ref030","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1002\/alz.13433","article-title":"Risk of cognitive decline progression is associated to increased blood\u2010brain\u2010barrier permeability: a longitudinal study in a memory unit clinical cohort","volume":"20","author":"A Puig\u2010Pijoan","year":"2023","journal-title":"Alzheimer\u2019s Dementia"},{"key":"pone.0343142.ref031","first-page":"297512","article-title":"Triggers and effectors of oxidative stress at blood-brain barrier level: relevance for brain ageing and neurodegeneration","volume":"2013","author":"A-M Enciu","year":"2013","journal-title":"Oxid Med Cell Longev"},{"key":"pone.0343142.ref032","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1007\/978-1-60761-938-3_17","article-title":"Novel models for studying the blood-brain and blood-eye barriers in Drosophila","volume":"686","author":"RL Pinsonneault","year":"2011","journal-title":"Methods Mol Biol"},{"issue":"1","key":"pone.0343142.ref033","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.cell.2005.07.029","article-title":"moody encodes two GPCRs that regulate cocaine behaviors and blood-brain barrier permeability in Drosophila","volume":"123","author":"RJ Bainton","year":"2005","journal-title":"Cell"},{"issue":"3","key":"pone.0343142.ref034","doi-asserted-by":"crossref","first-page":"1886","DOI":"10.3390\/ijms24031886","article-title":"Optimization and technical considerations for the dye-exclusion protocol used to assess blood-brain barrier integrity in adult Drosophila melanogaster","volume":"24","author":"K Bhasiin","year":"2023","journal-title":"Int J Mol Sci"},{"key":"pone.0343142.ref035","first-page":"533","article-title":"Honeybee workers as models of aging.","volume-title":"Conn\u2019s handbook of models for human aging","author":"TP Quigley","year":"2018"},{"issue":"2","key":"pone.0343142.ref036","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0148242","article-title":"Multi-drug resistance transporters and a mechanism-based strategy for assessing risks of pesticide combinations to honey bees","volume":"11","author":"AJ Guseman","year":"2016","journal-title":"PLoS One"},{"key":"pone.0343142.ref037","doi-asserted-by":"crossref","first-page":"1886","DOI":"10.3390\/ijms24031886","article-title":"Optimization and technical considerations for the dye-exclusion protocol used to assess blood\u2013brain barrier integrity in adult Drosophila melanogaster","volume":"24","author":"K Bhasiin","year":"2023","journal-title":"Int J Mol Sci"},{"issue":"11","key":"pone.0343142.ref038","doi-asserted-by":"crossref","first-page":"3538","DOI":"10.1523\/JNEUROSCI.5564-08.2009","article-title":"Evolutionary conservation of vertebrate blood-brain barrier chemoprotective mechanisms in Drosophila","volume":"29","author":"F Mayer","year":"2009","journal-title":"J Neurosci"},{"issue":"4","key":"pone.0343142.ref039","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0196749","article-title":"Relationship between brain plasticity, learning and foraging performance in honey bees","volume":"13","author":"A Cabirol","year":"2018","journal-title":"PLoS One"},{"issue":"9","key":"pone.0343142.ref040","doi-asserted-by":"crossref","first-page":"1322","DOI":"10.1002\/glia.21147","article-title":"Physiologic and anatomic characterization of the brain surface glia barrier of Drosophila","volume":"59","author":"MK DeSalvo","year":"2011","journal-title":"Glia"},{"key":"pone.0343142.ref041","doi-asserted-by":"crossref","first-page":"825695","DOI":"10.3389\/fncel.2022.825695","article-title":"Glial tiling in the insect nervous system","volume":"16","author":"N Pogodalla","year":"2022","journal-title":"Front Cell Neurosci"},{"issue":"3","key":"pone.0343142.ref042","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1523\/JNEUROSCI.4367-07.2008","article-title":"Organization and function of the blood-brain barrier in Drosophila","volume":"28","author":"T Stork","year":"2008","journal-title":"J Neurosci"},{"issue":"2","key":"pone.0343142.ref043","first-page":"232","article-title":"Dynamic analysis of the mesenchymal-epithelial transition of blood-brain barrier forming glia in Drosophila","volume":"6","author":"T Schwabe","year":"2017","journal-title":"Biol Open"},{"issue":"29","key":"pone.0343142.ref044","doi-asserted-by":"crossref","first-page":"17269","DOI":"10.1073\/pnas.2003909117","article-title":"Dynamic neural and glial responses of a head-specific model for traumatic brain injury in Drosophila","volume":"117","author":"J Saikumar","year":"2020","journal-title":"Proc Natl Acad Sci U S A"},{"key":"pone.0343142.ref045","doi-asserted-by":"crossref","first-page":"1","DOI":"10.7554\/eLife.85410","article-title":"Neural mechanisms of parasite-induced summiting behavior in \u2018zombie\u2019 Drosophila","volume":"12","author":"C Elya","year":"2023","journal-title":"eLife"},{"issue":"11","key":"pone.0343142.ref046","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0026796","article-title":"Killing them with kindness? In-hive medications may inhibit xenobiotic efflux transporters and endanger honey bees","volume":"6","author":"DJ Hawthorne","year":"2011","journal-title":"PLoS One"},{"issue":"6","key":"pone.0343142.ref047","doi-asserted-by":"crossref","first-page":"2659","DOI":"10.1038\/s41380-022-01511-z","article-title":"The blood-brain barrier in aging and neurodegeneration","volume":"27","author":"EG Knox","year":"2022","journal-title":"Mol Psychiatry"},{"issue":"3","key":"pone.0343142.ref048","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1038\/s43587-021-00043-5","article-title":"Healthy aging and the blood-brain barrier","volume":"1","author":"WA Banks","year":"2021","journal-title":"Nat Aging"},{"key":"pone.0343142.ref049","first-page":"533","article-title":"Honeybee workers as models of aging.","volume-title":"Conn\u2019s handbook of models for human aging","author":"TP Quigley","year":"2018"},{"issue":"5","key":"pone.0343142.ref050","doi-asserted-by":"crossref","first-page":"1792","DOI":"10.1073\/pnas.1818371116","article-title":"Varroa destructor feeds primarily on honey bee fat body tissue and not hemolymph","volume":"116","author":"SD Ramsey","year":"2019","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"3","key":"pone.0343142.ref051","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1046\/j.1570-7458.2001.00905.x","article-title":"The effect of the ectoparasitic mite, Varroa destructor on adult worker honeybee (Apis mellifera) emergence weights, water, protein, carbohydrate, and lipid levels","volume":"101","author":"PL Bowen\u2010Walker","year":"2001","journal-title":"Entomologia Exp Applicata"},{"key":"pone.0343142.ref052","doi-asserted-by":"crossref","first-page":"103877","DOI":"10.1016\/j.ibmb.2022.103877","article-title":"Omics-based analysis of honey bee (Apis mellifera) response to Varroa sp. parasitisation and associated factors reveals changes impairing winter bee generation","volume":"152","author":"M Kunc","year":"2023","journal-title":"Insect Biochem Mol Biol"},{"issue":"2","key":"pone.0343142.ref053","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pbio.3003035","article-title":"Macrophage invasion into the Drosophila brain requires JAK\/STAT-dependent MMP activation in the blood-brain barrier","volume":"23","author":"B Winkler","year":"2025","journal-title":"PLoS Biol"},{"issue":"44","key":"pone.0343142.ref054","doi-asserted-by":"crossref","DOI":"10.1126\/sciadv.abh0050","article-title":"Brain inflammation triggers macrophage invasion across the blood-brain barrier in Drosophila during pupal stages","volume":"7","author":"B Winkler","year":"2021","journal-title":"Sci Adv"},{"key":"pone.0343142.ref055","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.cois.2019.08.012","article-title":"Honey bees as bioindicators of changing global agricultural landscapes","volume":"35","author":"TP Quigley","year":"2019","journal-title":"Curr Opin Insect Sci"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0343142","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T17:45:40Z","timestamp":1774633540000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0343142"}},"subtitle":[],"editor":[{"given":"Olav","family":"Rueppell","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2026,3,27]]},"references-count":55,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,3,27]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0343142","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,27]]}}}