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Proof of Concept study

Dive deeper into our proof-of-concept study with Karolina Skonieczna-Żydecka (Pomeranian Medical University in Szczecin) in our IMMEDIATE insight blog.

The gut-brain-microbiota axis (GBMA) is a bidirectional pathway, both neuronal and biochemical, between the intestinal barrier and the central nervous system. Central, but not the only role in brain-gut communication is played by the gut microbiota. The composition of the gut microbiota, and thus the functions play an important role in functional disorders of both the gastrointestinal tract and the brain, significantly influencing the formation of cognitive, emotional and behavioural functions. [1]


Modulating the Gut Microbiota: Impact on Health and Behaviour

The composition of the microbiome is shaped by many factors that include diet, lifestyle, exposure to stress, as well as co-morbidities and medication. Short-term dietary modulation, even for a few hours, has been shown to cause significant changes in the composition of the microbiota. Similarly, exposure to stress and activation of the hypothalamic-pituitary-adrenal axis are rapidly reflected in the composition of the gut microbiota [2]. Changes in the gut microbiota secondary to chronic exposure to stress can be a trigger for its manifestation in the form of somatic and psychiatric symptoms. The disruption of homeostasis and changes in the gut microbial ecosystem then generated initiates a number of inflammatory disorders in the gut along with an increased synthesis of neuro-competent metabolites that can negatively affect the enteric and central nervous system.

Modulations in this GBMA therefore have a significant impact on cognitive, emotional and behavioural functions [3]. Modulation of the gut microbiota with probiotics and dietary protocols have shown that they can significantly alter stress-induced behaviour. In addition, intake of selected probiotic strains improved anxiety and depressive behaviour in mice. It has also been documented that in humans, probiotics also show beneficial effects on mental health. Some probiotic strains have a particularly significant impact on mental health and are therefore called psychobiotics.

Some limitations are imposed on psychobiotics (probiotics) due to their standardisation and the stability of the product during the final shelf life. Long-term storage leads to a change in the balance between viable and non-viable bacteria, which may affect the clinical efficacy of the product [4]. Therefore, the use of so-called postbiotics, which contain bacterial metabolites or other bacteria-derived fragments, are seen as new solutions and alternatives to the use of standard probiotics. One of the new postbiotics of interest to researchers and clinicians is pasteurised Akkermansia muciniphila MucT (PAM) [5].

Investigating the Efficacy of Pasteurised Akkermansia Muciniphila (PAM)

Akkermansia muciniphila is an anaerobic, Gram-positive bacterium representing approximately 1% to 3% of the total intestinal microbiota. A. muciniphila is abundant in intestinal mucus where it degrades mucin and thus can contact epithelial cells more easily than other non-mucin-degrading bacteria. A. muciniphila uses mucins as a source of carbon and nitrogen so that it actively produces short-chain fatty acids such as acetate and propionate [6]. Recent studies indicate that A. muciniphila supplementation can improve clinical exponents of metabolic syndrome, obesity, diabetes and inflammatory bowel disease in animals. The universal mechanism involved in exerting the benefits from PAM supplementation is elevating gut barrier integrity. [7]

The primary objective of the Proof-of-Concept study is to investigate the efficacy of pasteurised A. muciniphila MucT (PAM) in reducing somatic and psychological symptoms of stress. Secondary objectives include assessing the effects of PAM on the microbiota, metabolome and immune and gut barrier integration markers; Doctors, nurses, midwives and paramedics are professionals who are extremely exposed to stress and irregular lifestyles, at high risk of occupational burnout, anxiety, depression and other psychosomatic disorders. The study group will consist of at least 200 active healthcare professionals working in high-stress departments (trauma wards, surgery, internal medicine, delivery room, etc.) in shifts.

The following inclusion criteria were adopted:

  1. Ages 18-70,
  2. A minimum of one year's work experience in an establishment
  3. Body mass index (BMI) ≥18.5 kg/m2 and ≤ 35 kg/m2,

The exclusion criteria are as follows:

  1. Diagnosis of autoimmune diseases, neurological diseases, immune disorders, thyroid diseases, inflammatory bowel diseases, irritable bowel syndrome and other functional gastrointestinal diseases, diabetes, cancer and/or IgE-mediated allergy;
  2. Psychiatric comorbidities, including mental retardation, organic brain dysfunction or addiction (except nicotine and caffeine), taking antipsychotic and antidepressant medication;
  3. Use of proton pump inhibitors;
  4. Use of antibiotics and/or probiotics 4 weeks prior to testing;
  5. Treatment with corticosteroids and/or metformin;
  6. Use of food supplements (except vitamin D);
  7. Gastrointestinal surgery within the last 6 months before or planned during the trial
  8. Use of medication in the treatment of IBS (including peppermint oil, bile acid binders);
  9. Lactose intolerance;
  10. Participation in another study within the last 30 days before and during the study;
  11. Specific restrictive diet (e.g. elimination, vegan, FODMAP, reduction) 3 months prior to testing;
  12. Significant changes in physical activity 4 weeks prior to the study;
  13. Pregnancy or lactation.

The randomised clinical trial will enrol 200 adult shift healthcare workers. The postbiotic intervention will be conducted for 3 months. The active ingredient, present in the product, has received marketing authorisation as a novel food [8], and its safety has been confirmed by a Scientific Opinion of the European Food Safety Authority [9].

Prior to the introduction of the intervention, a history will be taken, including basic clinical and socio-demographic data on the patient and his/her family. Recruited individuals will be interviewed three times (beginning, middle and end of the study) to assess their nutritional status using the bioimpedance method and their dietary and physical activity habits using questionnaire methods, FFQ and IPAQ respectively. Emotional state will be assessed using psychometric methods, and the occurrence of functional gastrointestinal disorders will be assessed using relevant questionnaire methods used in gastroenterology practice. Blood pressure and heart rate will be measured at each visit.

The study material will be faeces and whole blood. Faeces and blood will also be collected at each of 3 visits the material will be used to perform carbohydrate and lipid metabolism biomarkers, as well as to assess the concentration of DHEAS (Dehydroepiandrosterone-sulfate -a stress marker; an alternative to cortisol). Analysis of the faecal microbiome will be performed by next-generation sequencing. The faecal and serum metabolome and markers of intestinal barrier permeability will also be determined. Immunological phenotypes and functions of isolated peripheral blood mononuclear cells will be characterised using scRNAseq, scATAC-seq and CyTOF techniques. Quantification of inflammatory mediators in plasma will be performed using OLINK® technology.

In this way, an intervention study using a so-called novel food, i.e. a pasteurised Akkermansia muciniphila bacterium with strong immunocompetent properties, will allow to assess the effectiveness of a non-invasive therapy for homeostasis of clinical biomarkers and maintenance of well-being.

  1. Skonieczna-Żydecka, K.; Marlicz, W.; Misera, A.; Koulaouzidis, A.; Łoniewski, I. Microbiome—The Missing Link in the Gut-Brain Axis: Focus on Its Role in Gastrointestinal and Mental Health. J Clin Med 2018, 7, 521, doi:10.3390/jcm7120521.
  2. Foster, J.A.; Rinaman, L.; Cryan, J.F. Stress & the Gut-Brain Axis: Regulation by the Microbiome. Neurobiol Stress 2017, 7, 124–136, doi:10.1016/j.ynstr.2017.03.001.
  3. Drossman, D.A.; Hasler, W.L. Rome IV-Functional GI Disorders: Disorders of Gut-Brain Interaction. Gastroenterology 2016, 150, 1257–1261, doi:10.1053/j.gastro.2016.03.035.
  4. Szajewska, H. QUALITY OF PROBIOTIC PRODUCTS: WHAT IS NEEDED? Journal of Pediatric Gastroenterology and Nutrition 2016, 63, S43, doi:10.1097/01.mpg.0000489605.04419.55.
  5. Derrien, M.; Vaughan, E.E.; Plugge, C.M.; de Vos, W.M. Akkermansia Muciniphila Gen. Nov., Sp. Nov., a Human Intestinal Mucin-Degrading Bacterium. Int J Syst Evol Microbiol 2004, 54, 1469–1476, doi:10.1099/ijs.0.02873-0.
  6. Cani, P.D.; de Vos, W.M. Next-Generation Beneficial Microbes: The Case of Akkermansia Muciniphila. Front Microbiol 2017, 8, 1765, doi:10.3389/fmicb.2017.01765.
  7. Liu, Y.; Liu, Q.; Zhang, C.; Zhao, J.; Zhang, H.; Chen, W.; Zhai, Q. Strain-Specific Effects of Akkermansia Muciniphila on the Regulation of Intestinal Barrier. Food Science and Human Wellness 2023, 12, 1526–1537, doi:10.1016/j.fshw.2023.02.022.
  8. Commission Implementing Regulation (EU) 2022/168 of 8 February 2022 Authorising the Placing on the Market of Pasteurised Akkermansia Muciniphila as a Novel Food under Regulation (EU) 2015/2283 of the European Parliament and of the Council and Amending Commission Implementing Regulation (EU) 2017/2470 (Text with EEA Relevance); 2022; Vol. 028;
  9. Safety of Pasteurised Akkermansia Muciniphila as a Novel Food Pursuant to Regulation (EU) 2015/2283 | EFSA Available online: (accessed on 22 January 2023).