Use of GutAlive and its impact on the standardization of downstream microbiota-based studies

Trabajo presentado en el 10th Workshop on Probiotics and Prebiotics, de la Sociedad Española de Probióticos y Prebióticos (SEPyP), celebrado en Las Palmas de Gran Canarias, del 6 al 8 de febrero de 2019

[Aim] The aim of this work was to evidence that the use of different stool collection strategies has a deep impact on the viability and diversity of the fecal microbiota that is recovered in the laboratory, notably if oxygen toxicity is not considered. This is particularly critical if the purpose of the sample is to envisage personalized biotherapeutic purposes, such as autologous fecal microbiota transplant or designing personalized biotherapeutics. [Methods] In order to analyze differences in the viability of fecal microbial populations during the whole delivery procedure, the same fecal specimens were sampled in conventional stool containers and GutAlive® devices, which minimize exposure of fecal microbiota to oxygen. Samples from five healthy donors were used and 150 differential colonies between the two devices were recovered. Differences included those associated to the two collection devices and individuals. All colonies were identified by 16S rRNA gene sequencing. Complete genomes of two extremely oxygen sensitive (EOS) bacteria recovered were sequenced with the Illumina MiSeq Sequencing System. [Results] Microbial diversity obtained was notably higher using GutAlive®. This device was able to maintain the viability of EOS species such as Akkermansia muciniphila, Faecalibacterium prausnitzii and a possible new member of the Clostridiales order. These obligate anaerobes were not recovered using the conventional stool container. Remarkably, GutAlive® allowed culturing and identifying an anaerobic isolate which may represent a new lineage within Clostridiales. This shows the importance of a personalized approach in microbiota-based therapies, as this novel isolate was recovered only from feces of one of the donors. [Conclusion] Using GutAlive® for stool sampling and transport allowed higher recovery of viable EOS bacteria by limiting oxygen exposure during the whole process. By standardizing the sampling and transport of the fecal specimens to the lab, GutAlive® can be applied for the normalization of microbiota-based studies, analysis and developments. Competing interests: NM is a full-time employee of Microviable Therapeutics; CH-C, SD, AM and BS are co-founders and SAB members of Microviable Therapeutics, and also co-inventors on at least one patent regarding microbiome stool collection kit and applications.