AUTHOR=Hamel Darbandi Marvie , Dalmacio Leslie Michelle M. 
  
TITLE=Differential in vitro and in vivo responses of Akkermansia muciniphila to Odontosoria biflora (Kaulf.) C.Chr. [Lindsaeaceae] hexane extract in diet- and alloxan-induced BALB/c mice
  
JOURNAL=Acta Biochimica Polonica
  
VOLUME=Volume 73 - 2026
  
YEAR=2026
  
URL=https://www.frontierspartnerships.org/journals/acta-biochimica-polonica/articles/10.3389/abp.2026.16199
  
DOI=10.3389/abp.2026.16199
  
ISSN=1734-154X
  
ABSTRACT=Akkermansia muciniphila is a mucin-degrading gut bacterium linked to metabolic health, yet culture-based growth stimulation may not translate to sustained enrichment in vivo. Odontosoria biflora (“tubho”) is traditionally consumed in the Philippines as a herbal tea, but its extract-level activity toward A. muciniphila remains poorly characterized. O. biflora was sequentially extracted (hexane, ethyl acetate, methanol, aqueous) and screened for growth-supporting activity toward A. muciniphila in modified BHI under anaerobic conditions. The most active fraction (hexane; OBE HEX) was evaluated for acute oral tolerability in BALB/c mice according to OECD Tesy No. 423 (up to 2000 mg/kg) and subsequently assessed in a high-fat/high-sugar diet plus alloxan-induced diabetic model. Fecal A. muciniphila-specific qPCR signal was monitored at weeks 0, 1, 2, and 4 using a modified 2−ΔΔCt approach with external ATCC genomic DNA as a reference. OBE-HEX produced the strongest in vitro growth-supporting effect (56.43% at 250 mg/L, p < 0.05; 85.62% at 500 mg/L, p < 0.001) and showed no observable toxicity in vivo. In contrast, in vivo analysis revealed only transient changes in fecal A. muciniphila detection following OBE-HEX administration, whereas sustained elevation was observed only in metformin-treated mice. Untargeted UPLC-ESI-QTOF-MS analysis of OBE-HEX yielded putative identification of 2-O-rhamnosylvitexin and 7-methoxy-9,10-dihydrophenanthrene-2,5-diol. Overall, these findings demonstrate that while O. biflora hexane extract exhibits direct growth-supporting activity toward A. muciniphila in vitro and is orally tolerable, such effects do not translate into sustained in vivo enrichment under diabetic conditions, underscoring the limitations of extrapolating culture-based microbiota screening results to host-associated systems.