Collection Citation: Philip Laric, Armina Mortazavi, Ewa Węgrzyn, Kathrin Simon, Pauline Sophie Rittel, Florain Markus Trefz, Benedikt Sabass 2025. In vitro Cultivation and Microbiome Analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygxwrnkv8j/v1
Manuscript citation:
Laric PP, Mortazavi A, Węgrzyn E, Simon K, Rittel PS, Trefz FM, Sabass B (2025) A reliable in vitro rumen culture system and workflow for screening anti-methanogenic compounds. PLOS One 20(12). doi: 10.1371/journal.pone.0335844
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this collection and it's working
Created: February 20, 2025
Last Modified: October 24, 2025
Collection Integer ID: 124080
Keywords: In vitro rumen cultivation, Methane inhibition, Microbiome analysis, PCR, Next generation sequencing, Greenhouse gas, ruminants, Cow, functioning rumen cultivation setup, rumen cultivation setup to screen, rumen cultivation, other rumen cultivation, ruminal culture, analysis of the microbiome, microbiome analysis, microbiome, ruminal, microbiome analysis this protocol, cultivation
Funders Acknowledgements:
German Federal Ministry of Education and Research (BMBF)
Grant ID: 031B1504
European Union’s Horizon 2020 research and innovation programme
Grant ID: 852585
Abstract
This protocol describes the steps for building a setup for in vitro rumen cultivation. The full workflow of inoculation of ruminal cultures, measurement of produced gases, as well as the analysis of the microbiome is described. As result one should be able to build a fully functioning rumen cultivation setup to screen for compounds of interest or to conduct any other rumen cultivation related studies.
Materials
General equipment:
8-Channel pipette (Ergonomic High Performance multichannel pipette, VWR, Darmstadt, Germany)
