PREDISMET

Congresos

• Comunicación oral: 1. Anti-obesity properties of Latilactobacillus sakei in C. elegans and diet-induced obese Wistar rat models. 14th EUROPEAN NUTRITION CONFERENCE FENS 2023. 14/11/2023-17/11/2023, Sava center, Belgrado, Serbia, Federation of European Nutrition Societies (FENS).
• Comunicación oral: Novel Lactiplantibacillus plantarum and Lactobacillus alimentarius strains reduce fat accumulation and improve health markers on a C. elegans model, 20th CTPIOD meeting (Conference for Trans-Pyrenean Investigations on Obesity and Diabetes), 16/10/2023-17/10/2023, Bordeaux Institute of Technology (Bordeaux, France), Consortium for TransPyrenean investigations on Obesity and Diabetes
  (CTPIOD)
• Comunicación oral: THE PROBIOTIC LACTOBACILLUS PLANTARUM REDUCES FAT ACCUMULATION AND INCREASES HEALTHSPAN IN CAENORHABDITIS ELEGANS, XXXI Congreso de la Sociedad Española de Nutrición, 14/09/2023-17/09/2022, Cartagena, Murcia, Sociedad Española de Nutrición (SEÑ)
• Póster: The probiotic Lactobacillus sakei reduces fat accumulation, oxidative stress and senescence while increasing life span in Caenorhabditis elegans, VIII Spanish Worm Meeting, 21/10/2022-22/10/2022, Logroño, La Rioja, Spanish Worm Society
• Comunicación oral: Pipeline de trabajo a la hora de identificar y trabajar con pro/para/postbioticos in vitro, en animales y humanos, XIV Workshop de la Sociedad Española de Microbiota Probióticos y Prebióticos (SEMiPyP), 08/03/2023-10/03/2023, Pamplona, Navarra, Sociedad Española de Microbiota Probióticos y Prebióticos.

The general goal of the PREDISMET project (2020-2022 collaborative R&D), was to identify and characterise, using different approaches, bacterial strains or their metabolites that, either in isolation or in combination with probiotic compounds, are capable of reverting alterations in the composition of the intestinal microbiome (dysbiosis) that characterises illnesses related with metabolic syndromes like obesity, and to determine their safety.

The final goal of the project was to, in the near future, be able to bring about the development and industrial use of probiotic, prebiotic or symbiotic agents for the treatment of the metabolic alterations of diseases like excessive adiposity, insulin resistance, dyslipidemia or fatty liver disease.

The consortium between CNTA and CIN-UNAV obtained the following results.

1. Initially, CNTA worked on selecting candidate bacterial species to be developed as probiotic agents. To those ends, they did an exhaustive tuning of several tests that measure in vitro probiotic aptitude: antibiotic resistance tests, tests for resistance to gastric and intestinal stimulants, beta-galactosidase activity, bile salt hydrolase capacity, short-chain fatty acid production, intestinal epithelial bacterial adhesion, immunomodulator capacity, activities on cellular nitric oxide production and in vitro lipolytic activity. The entire battery of tests made it possible to select three Lactobacillus strains (A, P and S) as potentially probiotic strains for subsequent functional evaluation.
2. CNTA developed a colon simulator in which the counts of three indicator bacteria (selected as representative of the intestinal microbiome) were kept stable in a discontinuous in vitro fermenting system for 19 hours. In addition, a molecular tool (q-PCR) was developed to monitor the indicator strains of the in vitro colonic fermentation system.
3. A mixture of borage seed oil (rich in omega-6 polyunsaturated fatty acids) and a quercetin polyphenol as an ingredient with prebiotic activity was selected because of its capacity to increase bacterial diversity in rodents.
4. The CIN-UNAV did an in vivo screening (C. elegans model) of potential anti-obesogenic activity of the bacterial strains selected by CNTA as potential probiotics. This study made it possible to show that the three Lactobacillus strains (A, P and S) selected were capable of lowering the body fat of nematodes, lengthening their life expectancy, reducing oxidative stress and slowing ageing. In addition, it was shown that the various properties observed were independent of an effect on the development of C. elegans, in addition to using mechanisms that encompass insulin signalling and the beta-oxidation of fatty acids. These studies led to the selection of two of the three Lactobacillus strains (P and S) for functional evaluation in a rodent model.
5. CNTA fine-tuned the way of obtaining postbiotics of bacterial origin for the three candidate strains by isolating the cell-free supernatant (CFS) resulting from in vitro probiotic growth. In vitro (cell cultures) and in vivo (C elegans) screening of those CFS made it possible to identify their anti-lipogenic and lipolytic activity.
6. The metabolomic analysis not aimed at the CFS of the three bacterial strains, in addition to other strains from the CNTA microorganism collection, made it possible to identify different metabolites produced by different bacteria.  One of those metabolites, found in the CFS of the S strain of Lactobacillus, was confirmed using a commercial pattern.
7. The in vitro screening using metabolite cell lines identified and confirmed in the CFS of the Strain of Lactobacillus showed anti-lipogenic and antiadipogenic properties in HepG2 hepatocytes and 3T3-L1 MBX preadipocytes, respectively.
8. CNTA carried out the in vitro colonic fermentation study to determine the effect of the addition of prebiotic ingredients to the bacterial growth. They worked with the addition of 0.8% of a short-chain Fructooligosaccharide (FOS) with interesting properties for growing beneficial bacteria.
9. CIN-UNAV carried out the functional evaluation of the probiotic strains selected (Lactobacillus P and S) in combination with a prebiotic mixture of borage seed oil and quercetin (synbiotics) in a rat model with diet induced obesity. The study showed that supplementing with the probiotic strains, both freely and combined with the prebiotic ingredient, showed a significant reductive effect on the accumulation of adipose tissue, glycaemia control, or inflammation markers, among others.
10. The metagenomic study done by 16S sequencing of animal faeces from the different groups showed that the probiotics significantly affected intestinal microbiome composition, in terms of both bacterial species and diversity.
11. Undirected metabolomic studies were done from animal serum from the different groups that are making it possible to identify changes in the profile of metabolites associated with each probiotic or synbiotic.
12. Lastly, the UNAV performed a toxicological study of a Wistar rat model, which showed the safety of the Lactobacillus strains and prebiotic chosen.

Consequently, it can be said that the PREDISMET project made it possible for both CNTA and CIN-UNAV to obtain a large quantity of information about the properties of two Lactobacillus bacteria strains that, both freely and combined with a prebiotic ingredient, show potential to be developed as possible probiotic or synbiotic agents with effectiveness against parameters related with metabolic syndrome, like obesity or insulin resistance.