Guidelines for reporting on animal fecal transplantation (GRAFT) studies: recommendations from a systematic review of murine transplantation protocols

Abstract

Fecal microbiota transplant (FMT) is a powerful tool used to connect changes in gut microbial composition with a variety of disease states and pathologies. While FMT enables potential causal relationships to be identified, the experimental details reported in preclinical FMT protocols are highly inconsistent and/or incomplete. This limitation reflects a current lack of authoritative guidance on reporting standards that would facilitate replication efforts and ultimately reproducible science. We therefore systematically reviewed all FMT protocols used in mouse models with the goal of formulating recommendations on the reporting of preclinical FMT protocols. Search strategies were applied across three databases (PubMed, EMBASE, and Ovid Medline) until June 30, 2020. Data related to donor attributes, stool collection, processing/storage, recipient preparation, administration, and quality control were extracted. A total of 1753 papers were identified, with 241 identified for data extraction and analysis. Of the papers included, 92.5% reported a positive outcome with FMT intervention. However, the vast majority of studies failed to address core methodological aspects including the use of anaerobic conditions (91.7% of papers lacked information), storage (49.4%), homogenization (33.6%), concentration (31.5%), volume (19.9%) and administration route (5.3%). To address these reporting limitations, we developed the Guidelines for Reporting Animal Fecal Transplant (GRAFT) that guide reporting standards for preclinical FMT. The GRAFT recommendations will enable robust reporting of preclinical FMT design, and facilitate high-quality peer review, improving the rigor and translation of knowledge gained through preclinical FMT studies.

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“…The fecal transplant was performed based on an established protocol 45 . Briefly, the donor mice of EE were administrated with EE for 4 weeks.…”

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“…46 Finally, we and others have highlighted the need to standardize procedures and reporting of any studies involving the gut microbiota and microbiome and generated guidelines as a result. [47][48][49] The guidelines for reporting on animal fecal transplantation recommendations are particularly relevant to germ-free mice as they improve the rigor of experimental design involving FMTs. 48…”

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“…Particularly, Walter et al. noted that human microbiota-associated animal studies frequently fail to demonstrate recapitulation of disease-associated microbiota changes (or ‘dysbiosis’) in recipient rodents (<30%), while >80% of studies use pseudoreplication and are thus statistically underpowered to reach their conclusions [ 83 ], leading to guideline aimed at standardizing approaches to FMT in rodent models [ 84 , 85 ]. In addition to improving the rigor of these studies, Walter et al.…”