[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-0-en-105":3,"doc-seo-446620-105":59,"doc-detail-446620-en":130},{"code":4,"msg":5,"data":6},0,"success",[7,13,18,23,28,33,38,43,48,51,55],{"id":8,"doc_module":4,"doc_module_name":9,"category_name":10,"show_sort_weight":11,"slug":12},1,"Document","Story & Novel",90,"story-novel",{"id":14,"doc_module":4,"doc_module_name":9,"category_name":15,"show_sort_weight":16,"slug":17},2,"Literature",80,"literature",{"id":19,"doc_module":4,"doc_module_name":9,"category_name":20,"show_sort_weight":21,"slug":22},4,"Exam",70,"exam",{"id":24,"doc_module":4,"doc_module_name":9,"category_name":25,"show_sort_weight":26,"slug":27},5,"Comic",60,"comic",{"id":29,"doc_module":4,"doc_module_name":9,"category_name":30,"show_sort_weight":31,"slug":32},6,"Technology",50,"technology",{"id":34,"doc_module":4,"doc_module_name":9,"category_name":35,"show_sort_weight":36,"slug":37},7,"Healthcare",40,"healthcare",{"id":39,"doc_module":4,"doc_module_name":9,"category_name":40,"show_sort_weight":41,"slug":42},8,"Research & Report",30,"research-report",{"id":44,"doc_module":4,"doc_module_name":9,"category_name":45,"show_sort_weight":46,"slug":47},9,"Religion & Spirituality",20,"religion-spirituality",{"id":46,"doc_module":4,"doc_module_name":9,"category_name":49,"show_sort_weight":46,"slug":50},"World Cup","world-cup",{"id":52,"doc_module":4,"doc_module_name":9,"category_name":53,"show_sort_weight":52,"slug":54},10,"Lifestyle","lifestyle",{"id":56,"doc_module":4,"doc_module_name":9,"category_name":57,"show_sort_weight":24,"slug":58},19,"General","general",{"code":4,"msg":60,"data":61},"ok",{"site_id":62,"language":63,"slug":64,"title":65,"keywords":66,"description":67,"schema_data":68,"social_meta":123,"head_meta":125,"extra_data":127,"updated_unix":129},105,"en","evaluating-preservation-effects-on-honeybee-gut-microbiota-inocula","Evaluating preservation effects on honeybee gut microbiota inocula","","The honeybee (Apis mellifera) gut microbiota serves as a model for host–microbe studies, and fresh gut homogenates are often preferred for colonization but may be constrained by practical needs. This study tests how short-term storage conditions shape colonization efficiency and community integrity. Germ-free bees received gut homogenates either immediately, refrigerated for 1 or 3 days, or frozen for 7 days with or without glycerol. Survival was similar across groups, while 3-day refrigeration reduced gut mass. Absolute 16S rRNA quantification plus diversity and composition analyses showed broadly maintained total loads, but taxon-specific shifts, with Gilliamella especially sensitive to freezing. Short refrigeration best matched fresh community features, and the work emphasizes absolute quantification for accurate microbiome colonization evaluation.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":40,"@type":76,"position":81},"https://docshare.wps.com/document/research-report/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/evaluating-preservation-effects-on-honeybee-gut-microbiota-inocula/446620/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/evaluating-preservation-effects-on-honeybee-gut-microbiota-inocula/446620.png","ImageObject",300,407,{"name":92,"@type":93},"McGucket","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-09-30","2026-09-29",true,{"@type":102,"interactionType":103,"userInteractionCount":14},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"How were the preservation conditions applied to honeybee gut microbiota inocula?","Question",{"text":112,"@type":113},"Gut homogenates were prepared and either used immediately, refrigerated for one day or three days, or frozen for seven days with or without glycerol before feeding to germ-free bees.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"Did survival rates of inoculated bees differ across storage treatments?",{"text":117,"@type":113},"No significant differences in survival rates were observed among treatments.",{"name":119,"@type":110,"acceptedAnswer":120},"Why does the paper emphasize absolute quantification instead of relative abundance alone?",{"text":121,"@type":113},"Relative and absolute measurements showed discrepancies, and absolute quantification was necessary to accurately evaluate colonization and microbial load changes across preservation conditions.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},446620,1790741887,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":39,"category_name":40,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":14,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145},1236954412713,"https://us-avatar.wpscdn.com/davatar_29158cc5080c5b710cf443261637dec0","| Applied and Industrial Microbiology | Research Article  \nEvaluating preservation effects on honeybee gut microbiota inocula  \nKang Wang,1 Zixuan Xu,1 Jinmeng Ma,1 Ming Zheng,1 Yi Zhang,1 Zheguang Lin,1 Ting Ji1 AUTHOR AFFILIATION See affiliation list on p. 10.  \nABSTRACT The honeybee (Apis mellifera) gut microbiota serves as a model for host– microbe studies, as germ-free bees enable a relatively sterile system. Fresh gut homo genates are optimal for colonization, but immediate use may be limited by practical considerations, such as pathogen screening and the need to minimize repeated preparation for animal welfare. In this study, we systematically evaluated how short-term preservation conditions influence the colonization efficiency and structural integrity of the honeybee gut microbiota. Gut homogenates were prepared and either used immediately, refrigerated for one day or three days, or frozen for seven days with or without glycerol, before being fed to germ-free bees. Colonization outcomes were assessed by combining absolute quantification of bacterial 16S ribosomal RNA gene copy numbers with diversity indices and community composition analyses. Survival rates of inoculated bees did not differ significantly among treatments, but bees receiving inocula refrigerated for three days showed reduced gut mass, suggesting compromised microbial activity. Absolute quantification revealed that total bacterial loads were broadly maintained, yet taxon-specific responses varied, with Gilliamella showing marked sensitivity to freezing. Discrepancies between relative and absolute measurements highlighted the importance of absolute quantification for accurately evaluating colonization. Overall, inocula retained the ability to establish the characteristic core microbiota under all preservation conditions. Notably, refrigeration for one day preserved community features without any detectable differences from freshly preparedinocula, whereas longer refrigeration and freezing introduced measurable perturbations. These results provide methodological guidance for honeybee microbiota transplantation experiments and underscore the necessity of absolute quantification in microbiome research.  \nIMPORTANCE The gut microbes of honeybees are essential for their nutrition, immunity, and overall health. To study these microbes, scientists often use germ-free bees colonized with gut communities from donor bees. Fresh preparations are ideal, but repeated preparation is time-consuming, requires pathogen checks, and may involve sacrificing many bees. In this study, we tested practical preservation methods and found that both refrigerated and frozen samples retained the ability to colonize germ-free bees. However, one day of refrigeration preserved microbial abundance and composition most similar to fresh samples, whereas longer refrigeration and freezing introduced detectable changes. These findings provide researchers with clear guidance for preparing standardized microbial inocula while also minimizing repeated dissection of donor bees, thereby improving both experimental reproducibility and animal welfare.  \nKEYWORDS honeybees, gut microbiome, intestinal colonization, storage condition, absolute quantification  \nEditor John M. Chaston, Brigham Young University, Provo, Utah, USA  \nAddress correspondence to Ting Ji, [tji@yzu.edu.cn](tji@yzu.edu.cn). authors declare conflict of interest  \nH oneybees (Apis mellifera) are essential pollinators in agricultural and natural  \necosystems, providing critical services for crop production and biodiversity maintenance (1) . The honeybee gut microbiota has become a focal point of increasing research interest, given its critical roles in nutrition, immunity, pathogen defense, and the regulation of host behavior (2–4) . Adult worker bees harbor a relatively simple but highly specialized gut consortium dominated by Lactobacillus Firm-5, Bombilactobacillus Firm-4, Bifidobacterium, Snodgrassella alvi, and Gilliamella (5, ","cbCailsR0TaFu0rL","https://ap.wps.com/l/cbCailsR0TaFu0rL","pdf",983583,11,"English","# Abstract\n# Importance\n# Keywords","[{\"question\":\"How were the preservation conditions applied to honeybee gut microbiota inocula?\",\"answer\":\"Gut homogenates were prepared and either used immediately, refrigerated for one day or three days, or frozen for seven days with or without glycerol before feeding to germ-free bees.\"},{\"question\":\"Did survival rates of inoculated bees differ across storage treatments?\",\"answer\":\"No significant differences in survival rates were observed among treatments.\"},{\"question\":\"Why does the paper emphasize absolute quantification instead of relative abundance alone?\",\"answer\":\"Relative and absolute measurements showed discrepancies, and absolute quantification was necessary to accurately evaluate colonization and microbial load changes across preservation conditions.\"}]","Evaluating preservation effects on honeybee gut microbiota inocula | PDF",1790716324,28]