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Background Administering probiotic strains of Limosilactobacillus reuteri to poultry has been shown to improve poultry performance and health. Some strains of L. reuteri taxa can produce reuterin, a broad-spectrum antimicrobial compound from glycerol conversion, with high inhibitory activity against enterobacteria. However, little is known about the metabolism of glycerol in the complex chicken cecal microbiota nor the effect of glycerol, either alone or combined with L. reuteri on the microbiota. In this study, we investigated the effect of L. reuteri PTA5_F13, a high-reuterin-producing chicken strain and glycerol, alone or combined, on broiler chicken cecal microbiota composition and activity using the continuous PolyFermS model recently developed to mimic chicken cecal fermentation. Methods Three independent PolyFermS chicken cecal microbiota models were inoculated with immobilized cecal microbiota from different animals and operated continuously. The effects of two additional levels of glycerol (50 and 100 mM) with or without daily supplementation of chicken-derived L. reuteri PTA5_F13 (10 7  CFU/mL final concentration) were tested in parallel second-stage reactors continuously inoculated with the same microbiota. We analyzed the complex chicken gut microbiota structure and dynamics upon treatment using 16S rRNA metabarcoding and qPCR. Microbiota metabolites, short-chain and branched-chain fatty acids, and glycerol and reuterin products were analyzed by HPLC in effluent samples from stabilized reactors. Results Supplementation with 100 mM glycerol alone and combined with L. reuteri PTA5_F13 resulted in a reproducible increase in butyrate production in the three modelled microbiota (increases of 18 to 25%). Glycerol alone resulted also in a reduction of Enterobacteriaceae in two of the three microbiota, but no effect was detected for L. reuteri alone. When both treatments were combined, all microbiota quantitatively inhibited Enterobacteriaceae , including in the last model that had very high initial concentrations of Enterobacteriaceae . Furthermore, a significant 1,3-PDO accumulation was measured in the effluent of the combined treatment, confirming the conversion of glycerol via the reuterin pathway. Glycerol supplementation, independent of L. reuteri addition, did not affect the microbial community diversity. Conclusions Glycerol induced a stable and reproducible butyrogenic activity for all tested microbiota and induced an inhibitory effect against Enterobacteriaceae that was strengthened when reuterin-producing L. reuteri was spiked daily. Our in vitro study suggests that co-application of L. reuteri PTA5_F13 and glycerol could be a useful approach to promote chicken gut health by enhancing metabolism and protection against Enterobacteriaceae .

Quantitative wood anatomy is critical for establishing climate reconstruction proxies, understanding tree hydraulics, and quantifying carbon allocation. Its accuracy depends upon the image acquisition methods, which allows for the identification of the number and dimensions of vessels, fibres, and tracheids within a tree ring. Angiosperm wood is analysed with a variety of different image acquisition methods, including surface pictures, wood anatomical micro-sections, or X-ray computed micro-tomography. Despite known advantages and disadvantages, the quantitative impact of method selection on wood anatomical parameters is not well understood. In this study, we present a systematic uncertainty analysis of the impact of the image acquisition method on commonly used anatomical parameters. We analysed four wood samples, representing a range of wood porosity, using surface pictures, micro-CT scans, and wood anatomical micro-sections. Inter-annual patterns were analysed and compared between methods from the five most frequently used parameters, namely mean lumen area ( ), vessel density ( ), number of vessels ( ), mean hydraulic diameter ( ), and relative conductive area ( ). A novel sectorial approach was applied on the wood samples to obtain intra-annual profiles of the lumen area ( ), specific theoretical hydraulic conductivity ( ), and wood density ( ). Our quantitative vessel mapping revealed that values obtained for hydraulic wood anatomical parameters are comparable across different methods, supporting the use of easily applicable surface picture methods for ring-porous and specific diffuse-porous tree species. While intra-annual variability is well captured by the different methods across species, wood density ( ) is overestimated due to the lack of fibre lumen area detection. Our study highlights the potential and limitations of different image acquisition methods for extracting wood anatomical parameters. Moreover, we present a standardized workflow for assessing radial tree ring profiles. These findings encourage the compilation of all studies using wood anatomical parameters and further research to refine these methods, ultimately enhancing the accuracy, replication, and spatial representation of wood anatomical studies.