Laboratory handling practice for faecal microbiota transplantation

Abstract

Aims Faecal microbiota transplantation (FMT) consists of the infusion of faeces from a healthy donor to the gastrointestinal tract of a recipient patient to treat disease associated with alterations in gut microbiota. The objective of this article was to describe laboratory workflow of an FMT laboratory to provide tips for preparing the faecal suspensions to be infused. Methods and Results Twenty‐stool solutions obtained from ten donors were prepared using two different protocols: magnet plate emulsion (MPE) and Seward StomacherTM Emulsion (SSE). We evaluated parameters such as preparation time, handiness, and aerobic and anaerobic microbial count. For three donors, we monitored bacterial counts after defrosting at different time‐points. MPE requires more time than SSE. In terms of microbial load, both methods showed similar values, with small and statistically differences (P ≤ 0·05) regarding anaerobes in favour of SSE. Frozen aliquots showed the same bacterial load values after defrosting. Conclusion Although both methods allow an easy and available preparation of a stool suspension, SSE seems more suitable, particularly for stool banking. Aerobic and anaerobic species are preserved with both protocols; and safety for laboratory operators is guaranteed. Significance and Impact of the Study In recent years, FMT has become a fascinating and interesting subject. Nevertheless, there are no real guidelines describing laboratory facilities and procedures. This paper aims to be a useful and simple guide to increase the number FMT centres as much possible.

“Bacterial Consortium”: A Potential Evolution of Fecal Microbiota Transplantation for the Treatment of Clostridioides difficile Infection

“…In addition, these donors must result negative for a series of serological and microbiological screening tests [ 8 ]. Moreover, stool sample must be processed within six hours after defecation in order to preserve anaerobes [ 8 , 10 ]. All these aspects make the procedure laborious and poorly standardized.…”

“…FMT has shown interesting and promising results about the efficacy and management of the patients. But, concerning on procedural safety, potential pathogen transmission, and standardizing workflow, some key points have to be improved [ 10 ]. Recruiting of voluntary donors is a very critical aspect in FMT flowchart.…”

“…At first, each donor undergoes a general questionnaire to exclude intestinal and extraintestinal disorders. Subsequently, on donor stool and blood sample are performed cultural, molecular, and serologic analyses in order to avoid any possible transmission during infusion procedure [10][11][12]. It is clear that during screening process, a donor may result as unsuitable and, then, his fecal matter is unusable for the therapy.…”

Rummeliibacillus suwonensis: First Time Isolation from Human Feces by Culturomics

“…In detail, thirty grams of fecal sample were suspended in 30 mL of saline solution. After the homogenization by Stomacher® 400 Circulator (SEWARD, UK), the fecal suspension is split in two aliquots and centrifuged at 3500×g for 10 min [9]. Supernatants were discarded and the two pellets were resuspended: one in 15 mL of Rumen fluid and the other one in 15 mL of supplemented Brucella Broth (BB, Remel INC., Lenexa, USA).…”

Oral Microbiota: Discovering and Facing the New Associations with Systemic Diseases

“…Some new approaches, which involve the use of microorganisms belonging to the commensal bacterial flora, have been presented for the treatment of infections resistant to some antibiotics. Among them, the fecal microbiota transplantation approach consists of the infusion of feces from a healthy donor to the gastrointestinal tract of a recipient patient to treat disease associated with alterations in gut microbiota, thus also reducing other systemic diseases associated with that dysbiosis [25].…”