{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T07:08:47Z","timestamp":1789456127774,"version":"build-2803163510"},"reference-count":97,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T00:00:00Z","timestamp":1789430400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"\n Introduction<\/jats:title>\n Fresh rumen fluid is a promising donor material for rumen fluid transplantation (RFT), but poor storage stability and batch variability limit its use.<\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n In this study, lyophilized rumen fluid samples from goats fed alfalfa hay or corn stover diets were designated AHP and CSP, respectively. For the\n in vitro<\/jats:italic>\n fermentation assay, portions of lyophilized rumen fluid from each batch within each dietary group were pooled and reconstituted; the substrate-supplemented groups were designated AH and CS, whereas AH-CON and CS-CON served as the corresponding substrate-free controls.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Lyophilization yields were 6.29 and 6.38%, and culturable carbohydrate-utilizing bacterial counts were 3.79 \u00d7 10\u2077 and 3.93 \u00d7 10\u2077 CFU\/g, respectively, with no significant differences between AHP and CSP. AHP contained higher propionate and valerate concentrations (\n p<\/jats:italic>\n &lt; 0.05), whereas CSP contained higher acetate and isovalerate concentrations and had a higher acetate-to-propionate ratio (A\/P;\n p<\/jats:italic>\n &lt; 0.05). Bacterial, fungal, and methanogen community structures differed significantly between groups (\n p<\/jats:italic>\n &lt; 0.01), and CSP showed higher bacterial and fungal Shannon indices (\n p<\/jats:italic>\n &lt; 0.01). AHP was significantly enriched in the bacterial genera\n Xylanibacter<\/jats:italic>\n ,\n Segatella<\/jats:italic>\n ,\n Prevotellaceae_UCG-001<\/jats:italic>\n , and\n Succinivibrio<\/jats:italic>\n ; the fungal genera\n Neocallimastix<\/jats:italic>\n ,\n Piromyces<\/jats:italic>\n ,\n Feramyces<\/jats:italic>\n , and\n Aspergillus<\/jats:italic>\n ; and the methanogen genera\n Methanoculleus<\/jats:italic>\n and\n Methanosphaera<\/jats:italic>\n . CSP was significantly enriched in the bacterial genera\n norank_f__F082<\/jats:italic>\n ,\n Rikenellaceae_RC9_gut_group<\/jats:italic>\n ,\n norank_f__Bacteroidales_BS11_gut_group<\/jats:italic>\n ,\n norank_f__[Eubacterium]_coprostanoligenes_group<\/jats:italic>\n ,\n Ruminococcus<\/jats:italic>\n ,\n Monoglobus<\/jats:italic>\n ,\n norank_f__UCG-010<\/jats:italic>\n , and\n Candidatus Saccharimonas<\/jats:italic>\n ; the fungal genera\n Orpinomyces<\/jats:italic>\n ,\n unclassified_f__Neocallimastigaceae<\/jats:italic>\n ,\n Cyllamyces<\/jats:italic>\n ,\n Caecomyces<\/jats:italic>\n ,\n Pecoramyces<\/jats:italic>\n ,\n Anaeromyces<\/jats:italic>\n ,\n Joblinomyces<\/jats:italic>\n , and\n Alternaria<\/jats:italic>\n ; and the methanogen genera\n Methanobrevibacter<\/jats:italic>\n and\n Methanolacinia<\/jats:italic>\n . Cross-domain correlation analysis showed predominantly positive associations among genera enriched within the same roughage-source group and negative associations between AHP- and CSP-enriched genera. Compared with their substrate-free controls, AH and CS showed increased gas production, bacterial crude protein (BCP), total volatile fatty acids (TVFA), acetate, propionate, and butyrate, and decreased pH (\n p<\/jats:italic>\n &lt; 0.05). CS showed higher pH, acetate, isovalerate, TVFA, and A\/P, whereas AH showed higher gas production, butyrate, and valerate (\n p<\/jats:italic>\n &lt; 0.01).\n <\/jats:p>\n <\/jats:sec>\n \n Discussion<\/jats:title>\n Overall, the roughage source in the donor diets influenced the fermentation characteristics and microbial composition of lyophilized rumen fluid. After reconstitution, both inocula fermented exogenous substrate and exhibited different fermentation profiles, supporting their potential as storable inocula for standardization in RFT.<\/jats:p>\n <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1905439","type":"journal-article","created":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T06:25:10Z","timestamp":1789453510000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Lyophilized rumen fluid from goats fed different roughage-based diets: fermentation characteristics, microbial communities, and post-reconstitution in vitro fermentation performance"],"prefix":"10.3389","volume":"17","author":[{"given":"Meila","family":"Na","sequence":"first","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University","place":["Hohhot, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenliang","family":"Guo","sequence":"additional","affiliation":[{"name":"College of Food and Agriculture, Hetao College","place":["Bayannur, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ran","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University","place":["Hohhot, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haidong","family":"Du","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Inner Mongolia Minzu University","place":["Tongliao, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenan","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Grassland Research of Chinese Academy of Agricultural Sciences","place":["Hohhot, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University","place":["Hohhot, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renhua","family":"Na","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University","place":["Hohhot, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,15]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"122233","DOI":"10.1016\/j.ijpharm.2022.122233","article-title":"Freeze-drying: a flourishing strategy to fabricate stable pharmaceutical and biological products","volume":"628","author":"Abla","year":"2022","journal-title":"Int. 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