WO2022101511A1 - Symbiotic composition as feed additive for piglets or sows and the use thereof - Google Patents

Abstract

Disclosed is a symbiotic composition for the treatment or prevention of dysbiosis in piglets during nursing and/or post-weaning, comprising, an effective therapeutic or preventative quantity of (i) Enterococcus faecium, or (ii) Bifidobacterium animalis lactis, or (iii) Clostridium butyricum, or (iv) Bifidobacterium crudilactis, as probiotic and one or more prebiotics, this composition being intended for administration to a pregnant or nursing sow and/or to a nursing and/or post-weaning piglet, in order to promote an anti-inflammatory environment in the intestine of the nursing and/or post-weaning piglet.

Description

SYMBIOTIC COMPOSITION AS A FEED ADDITIVE FOR PIGLETS OR SOWS AND THEIR USE

The present invention relates to a symbiotic composition for the treatment or prevention of dysbiosis in suckling and/or post-weaning piglets.

Intestinal dysbiosis is an imbalance of the intestinal microbiota that can generate an overgrowth of pathogens (eg E. coli) causing diarrhea. The microbiota is defined as all the microorganisms present in a given environment (definition from the Quebec office of the French language, 2008). This particular environment is called the microbiome. The imbalance of the intestinal microbiota results in a reduction in the diversity of bacterial populations and an excess of pathogenic bacteria in the microbiota. Conversely, a microbiota that is balanced in the distribution of the species of microorganisms that compose it is said to be in eubiosis.

The meat production sector is growing rapidly in the world, pork production is increasing steadily and represents one of the largest consumers of antibiotics in animal production.

On pig farms, weaning (stopping breastfeeding and switching to dry feed) is a period associated with a risk for young animals. A recurring problem when weaning animals is the increased risk of diarrhea resulting from the abrupt transition in diet. This abrupt transition can lead to intestinal dysbiosis.

Diarrhea can lead to weight loss, even death of the newborn in severe cases. Routine treatment as well as prophylactic procedures applied for diarrhea detected just after the birth of the piglet are often not very effective.

This transition period also generates in the animal, in particular in the piglet, a significant stress at the behavioral, nutritional and environmental level, responsible for a strong reduction in food consumption. It is therefore crucial to control this risk of diarrhea because it causes a considerable loss for breeders since it is often associated with a high mortality rate, weight loss, stunted growth, and treatment-related costs.

Currently this risk of diarrhea is controlled by the addition of bacteriostatic mineral additives (ZnO, CuSO ₄ ) in the diet of weaned piglets but, beyond the resistance already observed to these compounds, the practice of adding additives bacteriostatic minerals leads to environmental problems (contamination of water by minerals leached and contained in livestock effluents).

Adding therapeutic antibiotics to animal feed around weaning is also common practice. This treatment remains problematic because it induces the resistance of bacteria to antibiotics. In farm animals, indiscriminate use of antibiotics can also promote intestinal dysbiosis. Moreover, knowing that resistance to antibiotics is increasing more and more, particularly in farm animals, the European Commission has since ¹ January 2006 banned the use of antibiotics as growth promoters in animal feed.

Given the growth of global demography and the growing demand for meat in the world, it is therefore crucial to find alternative solutions to the use of antibiotics and mineral additives to reduce the incidence and severity of digestive problems. in weaned animals and, more generally, to support the sustainable growth of the meat industry.

In view of the above, alternative solutions to antibiotics and bacteriostatic minerals have been developed involving optimal use of probiotics in combination or not with prebiotics.

Generally, a combination of at least one probiotic with at least one prebiotic is called a synbiotic, when the prebiotic acts as a substrate in synergy with the probiotic to provide a positive effect on health, in particular on digestion. The digestion of proteins, sugars and fats in monogastric individuals, and in particular in pigs and poultry, is based first on digestion taking place in the stomach, which constitutes an acid environment leading to denaturation of the macromolecules to form a digesta. The digesta then arrives in the intestine and again undergoes hydrolysis there by the action of the pancreatic juice which contains several proteases and by the action of aminopeptidases and intestinal dipeptidases.

The absorption of the products of these hydrolyses takes place in the upper part of the small intestine and all the undigested products in the small intestine reach the large intestine where they are subjected to the action of microorganisms (intestinal microbiota) resulting in a fermentation.

There are many publications that confirm the indispensable role of the intestinal microbiota on the health of animals (and humans). The intestinal microbiota also interacts permanently with the immune system present in the intestinal wall. Some studies have also demonstrated the ability of probiotics and/or prebiotics to rebalance the intestinal microbiota and their usefulness on intestinal health through generally direct and short-term effects.

The World Health Organization (WHO) and the Food and Agriculture Organization of the United Nations (FAO) have defined probiotics as "living microorganisms which, when ingested in sufficient quantity , exert positive effects on health, beyond the traditional nutritional effects”. There are broader definitions which do not refer to food, and where the word "ingested" can be replaced by "administered".

Although people often think of bacteria and other microorganisms as harmful "germs", many are actually helpful. Some bacteria help digest food, destroy pathogenic cells or produce vitamins. Many of the microorganisms in probiotic products are the same or similar to microorganisms that naturally live endogenously in our bodies.

Without being exhaustive, probiotics include various microorganisms such as bacteria Bacillus coagulons, Bacillus subfilis, Bifidobacterium animalis lacfis, Lactobacillus rhamnosus, Bacillus licheniformis, Lactobacillus plantarum, Bifidobacterium thermophilum, Clostridium butyricum, Enterococcus faecium, Bifidobacterium crudilactis, Bifidobacterium mongoliense, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus helveticus, Lactobacillus crispatus, Lactobacillus pontis, Lactobacillus akajohnsonii, Bifidobacterium longei, Collins , Pediococcus acidilacticiici, Faecalibacterium prausnitzii, Coprococcus catus, Roseburia intestinalis, Anaerostipes butyraticus. The most common probiotic bacteria are those of the genera Lactobacillus and Bifidobacterium. Probiotics also include yeasts such as Saccharomyces boulardii (Sanders ME, “Probiotics, strains matter”, Functional foods & nutraceuticals magazine, 2007, 36-41).

Prebiotics, on the other hand, are substrates selectively utilized by host microorganisms that selectively stimulate the growth or activity of desirable microorganisms (Expert consensus document: The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics, Nature Reviews Gastroenterology & Hepatology, 2017, 14, 491-502, https://doi.org/!0.1038/nrgastro.2017.75).

Without being exhaustive, prebiotics include, among others, inulin, beta-glucan, 2'-Fucosyllactose (2FL), oligosaccharides such as galactooligosaccharides (GOS), fructooligosaccharides (FOS), mannanoligosaccharide (MOS), xylooligosaccharide (XOS) , polyunsaturated fatty acids (polyunsaturated fatty acid: PUFA), conjugated linoleic acid (CLA).

A formulation combining at least one probiotic in combination with at least one prebiotic to prevent and treat bacterial infections that can lead to diarrhea, increase the weight of the animal and promote growth is known from document WO2014049023. The document WO2014049023 describes products and compositions which can be beneficial in breeding. Said products and compositions described therein comprise microorganisms, such as bacteria, in particular probiotic bacteria. The strains, as well as the compositions comprising them, can be administered to animals, preferably to farm animals, such as pigs. The administration can take place in the first days of life. According to this document, the administration of the products or compositions promote animal growth and animal weight gain. Bacterial infections can also be prevented or treated by said compounds or compositions. This document WO2014049023 mentions that a composition comprising at least 2 probiotics is suitable for treating diarrhea in newborn (unweaned) piglets. In particular, a composition comprising Lactobacillus reuteri and Enterococcus faecium makes it possible to reduce diarrhoea, to increase the average daily gain in weight gain and to reduce the mortality of the newborn piglet.

Document WO2018002671 discloses a composition for the treatment and/or nutrition of poultry such as broilers comprising (i) a commensal probiotic chosen from Bifidobacterium animalis, Collinsella tanakaei, Lactobacillus reuteri, Anaerostipes, Lactobacillus crispatus, Pediococcus acidilacticiici, Lactobacillus pontis , Faecalibacterium prausnitzii, Coprococcus catus, Roseburia intestinalis, Anaerostipes butyraticus, Butyricicoccus, Lactobacillus johnsonii and Ruminococcus sp.; and (ii) a prebiotic. This document also discloses the use of such a composition for the treatment of enteric diseases in poultry, such as necrotic enteritis.

The document WO2006133472A1 relates to a food additive and/or additive for animal or human drinking water promoting the health or the growth of probiotics. This document also relates to a use of the human and animal food additive and/or drinking water, in particular for the prevention of the harmful effect of a certain number of undesirable germs in the digestive system of animals and/or domestic birds.

Finally, the document WO2017156548A1 relates to certain foods comprising a fermentable nutritional component and a probiotic component, where the probiotic component is selected, on the basis of genetic and/or metabolic criteria, to specifically metabolize any free sugar monomer (Free Sugar Monomers (FSM )) or any free amino acid (Free Amino Acids (FAA)) or peptide that accumulate in the small and large intestines due to the fermentable nutritional component, which without this metabolization would be fermented and metabolized by bacteria less adaptive/opportunistic, creating blooms of deleterious gut bacteria and shifting the microbiome to a potential state of dysbiosis. Unfortunately, the known formulations are essentially used as a therapeutic agent for dysbiosis or microbiome imbalance in farm animals. The effects demonstrated in vivo by these formulations remain very general, such as weight gain or the percentage of animal mortality. Many probiotics are described in combination with any prebiotic for a wide variety of purposes. Moreover, according to Wang et al. Appl. Microbiol. Biotechnol. 2020, 104: 335-349, DOI: 10.1007/s00253-019-10259-6, it is accepted that symbiotic compositions comprising several probiotics will have an advantageous effect in combating dysbiosis compared to compositions comprising a single probiotic. On the other hand, such compositions comprising several probiotics have the disadvantage of being less simple and more expensive to produce industrially.

Furthermore, according to Barba-Vidal et al. Animal, 2018, 12, 2489-2498, DOI: 10.1017/S1751731 1 18000873, there is a glaring lack of reproducibility in the experiments showing the beneficial effect of probiotics.

Interventions at the end of the sow's gestation and/or from the birth of the piglet should allow, by the principle of early programming, to influence in an optimal and lasting way the good development of the microbiota of the newborn. Consequently, the positive effects for the animal's health would not only be observed at the start of life and during the 3-4 weeks after weaning, but they would also remain observable throughout adult life.

The first intestinal colonization of the piglet by the maternal microbiota at the time of farrowing is crucial for the establishment of a favorable intestinal microbiota of the newborn, but the composition of the microbiota can be influenced by many factors such as antibiotics, food, environment, infectious agents, etc.

The present invention therefore intends to overcome the aforementioned drawbacks by providing a composition comprising a therapeutically or preventively effective amount of

(i) Enterococcus faecium, or

(ii) Bifidobacterium animalis lactis, or (iii) Clostridium butyricum, or

(iv) Bifidobacterium crudilactis, as a probiotic, and one or more prebiotics, this composition being to be administered to a pregnant or lactating sow, and/or to a nursing and/or post-weaning piglet, to promote an anti -inflammatory in the intestine of said suckling and/or post-weaning piglet, with the aim of improving the health conditions of the newborn.

The present invention therefore aims to provide a symbiotic formulation which has an effect which is maintained over time by acting specifically on the modulation of the microbiota in an early manner, thus promoting intestinal eubiosis in a reliable manner. In addition, the present invention aims to provide a simple symbiotic formulation (symbiotic composition) with a determined main probiotic, which facilitates the production and verification of production batches. Indeed, the industrial production of a simple, minimal symbiotic composition will be easier, faster, and will be economically more profitable than the production of a symbiotic composition comprising, for example, several probiotics.

According to the present invention, it has thus been demonstrated that it was possible to provide a simple symbiotic formulation comprising as main probiotic Enterococcus faecium, or Bifidobacterium animalis lacfis, or Clostridium butyricum, or Bifidobacterium crudilactis which generates in a demonstrated manner and reproducible an anti-inflammatory environment in the digestive system, both of the pregnant animal and later of the newborn, which is maintained over time to protect the animal against subsequent dysbiosis.

The symbiotic formulation according to the present invention making it possible to modulate the microbiota of the fetus in a pregnant or newborn animal will make it possible to have a maximum prophylactic effect and will ensure optimal growth and development of the animal.

Indeed, the formulation according to the present invention promotes a healthy intestinal flora which thus protects the animal which ingests it, in particular when the latter is ingested by the pregnant female by the generation demonstrated and reproducible of an onti-inflammatory environment donations the stomach and/or the intestine, and this in a systematic way.

Indeed, it is crucial to analyze the impact of probiotics through 3 important criteria: the concentration of viable cells, the beneficial effect on the growth of the host and the beneficial effect on the functioning of the digestive tract thanks in particular to an anti-inflammatory impact (Markowiak P. and Slizewska K., Gut Pathog. 2018, 10, 21 , doi: 10.1 186/s 13099).

The symbiotic formulation according to the present invention provides an effective solution against dysbiosis and the problem of diarrhea occurring during weaning, of which the probiotic and the prebiotic(s) are chosen specifically not only on cell viability, but for their associated effects on the cell viability and on the anti-inflammatory response of the host microbiota.

It has in fact been demonstrated according to the present invention that the symbiotic formulation containing a probiotic and at least one prebiotic produces a reduction in the production of IL-8 in an in vitro test, and an increase in short chain fatty acids "Short Chain Fatty Acids, SCFAs” as well as an improvement in the diversity and quantity of endogenous bacterial populations such as Lactobacillus and bitidobacteria as well as bacteria involved in the metabolic pathways for the production of butyrate or other short-chain fatty acids. The probiotic and the prebiotic(s) together form a balanced gut microbiota, thereby leading to the generation together of an anti-inflammatory environment in the digestive system of the animal, in particular of the mammal, in particular pigs, preferably pregnant animals and newborns, in particular pregnant or non-pregnant sows and piglets.

Indeed, these indicators are important because it has been shown that an inflammatory environment of the gastrointestinal tract can generate intestinal dysbiosis (Zeng et al., Mucosal Immunol., 2017, 10,18-26, doi: 10.1038/mi.2016.75 ). It has also been shown that “Short Chain Fatty Acids, SCFA” may additionally have a beneficial effect of immunological regulators on the gastrointestinal environment (Parada Venegas et al., Frontiers in Immunology, 2019, 10, doi:l 0.3389 /fimmu.2019.00277). SCFAs are short chain fatty acids produced by the fermentation process in the colon. The acetate, the propionate and butyrate are the three main SCFAs. Butyrate is particularly important for the proper functioning of the colon because it is the main source of energy for colonocytes (colon epithelial cells). SCFAs can also reduce the pro-inflammatory response of gut epithelial cells (Iraporda C. et al., Immunology, 2015 10:1161-1169. https://doi.org/10.1016/j.imbio.2015.06. 004).

Administered to pregnant or non-pregnant sows and to piglets, the symbiotic formulation according to the present invention promotes the establishment of a balanced intestinal microbiota very early, protecting the pregnant pig or animal or even the newborn, more particularly the pregnant sow or the piglet in a sustained manner against dysbiosis by the probiotic and the prebiotic it contains, which together generate an anti-inflammatory environment.

Advantageously, the composition according to the invention is characterized in that the therapeutically or preventively effective amount of probiotic for the treatment or prevention of dysbiosis is an amount of probiotic for

(i) reduce diarrhea in suckling and/or post-weaning piglets, and/or

(ii) increase the growth of the suckling and/or post-weaning piglet.

Preferably, in the composition according to the invention, the therapeutically or preventively effective quantity for reducing diarrhea in suckling and/or post-weaning piglets is a quantity of probiotic according to which the quantity of soft and/or liquid stools produced by said suckling and/or post-weaning piglet is smaller than the quantity of soft and/or liquid stools produced by a control piglet during the same period of time and at the same stage of development.

Advantageously, the quantity therapeutically or preventively effective for increasing the growth of the suckling and/or post-weaning piglet is a quantity of probiotic according to which a) a weight of said suckling and/or post-weaning piglet is higher relative to the weight of a control piglet at the same stage of development and for the same period of time and/or b) a non-pathological growth curve said nursing and/or post-weaning piglet is above, for at least two days, a growth curve of a control piglet for the same period of time at the same stage of development.

In a preferred embodiment of the present invention, said one or more prebiotics is chosen from the group consisting of inulin, beta-glucan, 2' Fucosyllactose, GOS/FOS (galacto-oligosaccharide, fructo-oligosaccharide) , resistant starch and mixtures thereof.

According to the present invention, Bifidobacterium thermophilum comes from the Belgian Coordinated Collections of Microorganisms (BCCM/LMG Gent) listed under number 18892, Enterococcus faecium (LMG S-28935), Bifidobacterium animalis lactis is a strain deposited at the Belgian Coordinated Collections of Microorganisms (BCCM/LMG Gent) having the deposit number LMG P-28149, Clostridium butyricum (LMG1217), Bifidobacterium crudilactis is a strain deposited at the National Collection of Microorganism Cultures (CNCM, Institut Pasteur) having the deposit number ( CNCM 1-3342) as part of document WC2006122850.

Preferably, in the symbiotic composition according to the invention, the therapeutically or preventively effective amount of probiotic is between 1.00E+05 and 1.00E+015 CFU/day/animal, preferentially between 1.00E+06 and 1. 00E+013 CFU/day/animal, advantageously between 1.00E+07 and 1.00E+011 CFU/day/animal.

Advantageously, in the symbiotic composition according to the invention, an amount of said one or more prebiotics is between 0.1 and 1000 g/day/animal, preferentially between 0.5 and 100 g/day/animal, advantageously between 1 and 25 g/day/animal.

Preferably, in the symbiotic composition according to the invention, said probiotic consists for at least 80% by weight of a single probiotic chosen from the group consisting of (i) Enterococcus faecium, (ii) Bifidobacterium animalis lactis, (iii) Clostridium butyricum, and (iv) Bifidobacterium crudilactis. More preferentially, in the symbiotic composition, said probiotic consists for at least 85%, even more preferentially for at least 90%, advantageously for at least 95%, advantageously for at least 99%, 100% by weight of a single probiotic selected from the group consisting of (i) Enterococcus taecium, (ii) Bifidobacterium animalis lactis, (iii) Clostridium butyricum, and (iv) Bifidobacterium crudilactis.

Advantageously, the composition according to the invention is characterized in that it comprises a single probiotic chosen from the group consisting of (i) Enterococcus faecium, (ii) Bifidobacterium animalis lactis, (iii) Clostridium butyricum, and (iv) Bifidobacterium crudilactis.

Preferably, the composition according to the invention is in the form of a combination comprising said probiotic in a therapeutically or preventively effective amount and said one or more prebiotics, the combination being chosen from the group consisting of Bifidobacterium crudilactis and beta-glucans, Bifidobacterium crudilactis and inulin , Bifidobacterium crudilactis and 2'fucosyllactose, Bifidobacterium crudilactis and GOS/FOS, Bifidobacterium crudilactis and resistant starch, Bifidobacterium animalis lactis and 2'fucosyllactose, Bifidobacterium animalis lactis and inulin, Bifidobacterium animalis lactis and beta-glucans, Bifidobacterium animalis lactis and GOS /FOS, Bifidobacterium animalis lactis and resistant starch, Clostridium butyricum and GOS/FOS, Clostridium butyricum and inulin, Clostridium butyricum and beta-glucans, Clostridium butyricum and 2'fucosyllactose, Clostridium butyricum and resistant starch, Enterococcus taecium and beta -glucans, Enterococcus taeciu m and inulin, Enterococcus taecium and 2'fucosyllactose, Enterococcus taecium and GOS/FOS, Enterococcus taecium and resistant starch.

Advantageously, in the composition according to the invention, the combination is preferentially chosen from the group consisting of Clostridium butyricum and inulin, Clostridium butyricum and GOS/FOS, Bifidobacterium animalis lactis and inulin, Bifidobacterium animalis lactis and 2′fucosyllactose, Enterococcus taecium and beta -glucans, and Bifidobacterium crudilactis and beta-glucans, so as to allow a daily probiotic dosage of between 1.00E+05 and 1.00E+015 CFU/day/animal, preferably between 1.00E+06 and L00E+013 CFU/day/animal, advantageously between 1.00E+07 and 1.00E+01 1 CFU/day/animal, and a daily prebiotic dosage of between 0.1 and 1000 g/day/animal, preferably between 0 5 and 100 g/day/animal, advantageously between 1 and 25 g/day/animal. Indeed, according to the present invention, a symbiotic composition comprising for example Bifidobacterium crudilactis and beta-g lucanes, or comprising Bifidobacterium animalis lac fis and 2'fucosyllactose or comprising Clostridium butyricum and GOS/FOS or also comprising Enterococcus faecium and beta-g lucanes has shown a synergistic capacity to create an anti-inflammatory environment, by reducing the presence of IL-8, by increasing the production of short-chain fatty acids and promoting the growth of bacterial populations favorable to the microbiota.

Preferably, the composition according to the invention contains at least one conventional excipient, in liquid or solid form.

Preferably, the composition according to the invention is characterized in that said probiotic and/or said one or more prebiotics and/or said at least one conventional excipient are in encapsulated form(s) and/or in powder form and/or in the form of granules and/or in liquid form. Said probiotic and said one or more prebiotics of the composition are administered simultaneously, separately or staggered over time.

Preferably, the therapeutically or preventively effective amount provides treatment or prevention of dysbiosis for a period of time, counted from the day of birth of said piglet, greater than 3 days, preferably greater than 5 days, even more preferably greater than 15 days, favorably over 50 days, even more favorably over 100 days. Indeed, it has been shown that the composition according to the present invention can generate a persistence effect to treat or prevent dysbiosis in piglets. Depending on the symbiotic composition and the administration profile of this composition, for example depending on administration of the composition to the pregnant and/or lactating sow and/or to the lactating piglet, the treatment or prevention of piglet dysbiosis may be favored even when the piglet is at the post-weaning stage.

Preferably, the composition according to the invention is administered to a pregnant sow for the treatment or prevention of dysbiosis in suckling and/or post-weaning piglets. Alternatively, the composition according to the invention is administered to a suckling sow for the treatment or prevention of dysbiosis in suckling piglets and/or post-weaning. Alternatively, the composition according to the invention is administered to a suckling piglet for the treatment or prevention of piglet dysbiosis in breastfeeding and/or post-weaning. In an alternative preferred manner, the composition according to the invention is administered to a pregnant and/or lactating sow and/or to a lactating piglet for the treatment or prevention of dysbiosis in the piglet during lactation and/or post-weaning. .

Advantageously, the therapeutically effective amount for the treatment or prevention of dysbiosis is a therapeutically or preventively effective amount of probiotic to increase the production of mucus in the intestine of said suckling and/or post-weaning piglet according to which an amount of goblet cells (goblet cells) in the intestine of said suckling and/or post-weaning piglet is greater than the quantity of goblet cells in the intestine of said control piglet at the same stage of development.

Preferably, the therapeutically effective amount for the treatment or prevention of dysbiosis is a therapeutically or preventively effective amount of probiotic for increasing the absorption capacity of the intestinal villi of suckling and/or post-weaning piglets according to which

(i) a ratio between the depth of the intestinal villi and the height of the intestinal crypts in the intestine of said suckling and/or post-weaning piglet is greater than the ratio between the depth of the intestinal villi and the height of the intestinal crypts in the intestine of a control piglet at the same stage of development, and/or

(ii) a thickness of the intestinal lamina propria in the intestine of said suckling and/or post-weaning piglet is greater than the thickness of the intestinal lamina propria in the intestine of a control piglet at the same stage of development, and/or

Preferably, the therapeutically effective quantity for the treatment or prevention of dysbiosis is a quantity of probiotic according to which the concentrations of endogenous bacterial populations favorable to the intestinal microbiota such as lactobacillus and bifidobacteria are increased in the suckling piglet and/or in post-weaning compared to the concentrations of endogenous bacterial populations favorable to the intestinal microbiota of the control piglet at the same stage of development. Advantageously, this increase in population concentrations bacteria generates a better balance of the host's intestinal microbiota, reducing the risk of intestinal dysbiosis.

Advantageously, according to the invention, the therapeutically effective amount for the treatment or prevention of dysbiosis is a therapeutically effective amount of probiotic according to which the concentration of IL-8 is reduced by at least 10% in the intestine of the piglet in suckling and/or post-weaning relative to the concentration of IL-8 in the intestine of the control piglet at the same stage of development, said reduced concentration of IL-8 forming said anti-inflammatory environment. In the context of the present invention, the concentration of IL-8 was measured by means of in vitro tests on IPEC_J2 cells in the presence of the fermentation juice (containing or not containing the symbiotic composition to be tested).

Advantageously, according to the invention, the symbiotic composition is preferably characterized in that the therapeutically effective amount for the treatment or prevention of dysbiosis is a therapeutically effective amount according to which the secretion of short-chain fatty acids (SCFA's ) by the intestinal microbiota of a suckling and/or post-weaning piglet and/or of intestinal cells in vitro, is stabilized or increased compared to the secretion of short-chain fatty acids of a control piglet and/or or control intestinal cells in vitro. Said SCFAs, produced during the fermentation process of the microbiota are an indicator of the balance and growth of the microbiota. SCFAs have a multiple beneficial role in the intestine: butyrate, in particular, has a regulatory role in the transport of transepithelial fluid. It also improves the oxidative state and inhibition of inflammation of the intestinal mucosa, it strengthens the epithelial defense barrier, it modulates visceral sensitivity and intestinal motility (Canani, R. B. et al. World J. Gastroenterol., 2011, 17:1519-1528, doi:10.3748/wjg.v17.i12.1519).

Preferably, according to the invention, said SCFAs are chosen from butyrate, propionate, acetate, lactate or combinations thereof.

In the context of the present invention, the production of said SCFAs is measured using the in vitro static fermentation model detailed in more detail below. Advantageously, according to the invention, the synbiotic composition can contain more than 1 probiotic, for example 2 probiotics, or for example 3 probiotics, or for example 4 probiotics or for example 5 probiotics. According to the invention, the symbiotic composition may also contain more than 1 prebiotic, for example 2 prebiotics, or for example 3 prebiotics, or for example 4 prebiotics. Other embodiments of the symbiotic composition according to the invention are indicated in the appended claims.

The present invention also relates to a food supplement which can be administered to pregnant or non-pregnant sows and to piglets for the preventive and/or curative treatment of intestinal dysbiosis.

Said food supplement, comprising said composition, can be in liquid form, for example in a milky or oily base, or in solid form such as a powder, a powder in a capsule, a granule or a tablet or any other form whatsoever.

Other embodiments of the food supplement according to the invention are indicated in the appended claims.

The present invention also relates to a complete or complementary feed for farm animals comprising said composition, said feed being in the form of flour and/or granules and/or milk feeds and/or any other forms of food packaging. .

Other embodiments of the complete or complementary feed for livestock according to the invention are indicated in the appended claims.

The present invention further relates to a food for newborn animals comprising said composition according to the present invention and a milk base compatible with the diet of the newborn.

Other embodiments of the food for newborn animals according to the invention are indicated in the appended claims.

The present invention also relates to a use of the composition to be administered to a pregnant or lactating sow, and/or to a suckling and/or post-weaning piglet for the treatment or prevention of dysbiosis in suckling and/or post-weaning piglets in which the therapeutically or preventively effective amount of probiotic is between 1.00E+05 and L00E+015 CFU/day/animal, preferably between 1.00E+06 and 1. 00E+013 CFU/day/animal, advantageously between 1.00E+07 and 1.00E+01 1 CFU/day/animal, the amount of prebiotic is between 0.1 and 1000 g/day/animal, preferably between 0.5 and 100 g/day/animal, advantageously between 1 and 25 g/day/animal, the probiotic and the prebiotic of the composition being administered simultaneously, separately or staggered over time.

The present invention also relates to a use of the aforedescribed symbiotic composition, for the manufacture of a medicament for the treatment or prevention of dysbiosis in suckling and/or post-weaning piglets to be administered to a pregnant or lactating sow , and/or to a suckling and/or post-weaning piglet.

Other forms of use of said composition according to the invention are indicated in the appended claims.

The present invention finally relates to a method for treating or preventing dysbiosis in suckling and/or post-weaning piglets, comprising administration of the composition according to the invention to a pregnant or non-pregnant sow, for example a sow lactating piglet, and/or suckling piglet and/or post-weaning piglet.

Other characteristics, details and advantages of the invention will become apparent from the description given below, on a non-limiting basis and with reference to the drawings and the examples.

Figure 1 shows the schematic of the experimental device of the BabySPIME system (Dufourny S. et al. Journal of Microbiological Methods, 2019, 167: 105735. https://doi.Org/10.1016/j.mimet.2019). Compartment 1:

Stomach/duodenum/jejunum, compartment 2: ileum, compartment 3: ascending colon. A: Food, B: probiotic, C: prebiotic.

Figure 2 illustrates the level of IL-8 production (pg/ml) by IPEC-J2 cells. The IPEC-J2 cells were brought into contact with a fermentation juice coming from a fermentation with only a prebiotic: GOS/FOS (GF) or Inulin (Inu) or 2'FL (2FL) or beto-glucone (BG) or resistant starch (RS), or else originating from a fermentation during which a composition combining a prebiotic and a probiotic according to the invention was tested. The results show the mean of the level of IL-8 production +/- the standard deviation. The horizontal line corresponds to the level of IL-8 in the control condition (blank-mucin). Statistical significance was analyzed by a one-way ANOVA test with a Dunnett-type multiple comparison of each prebiotic and symbiotic against the control condition (indicated by *), and between the symbiotic combinations and the prebiotic alone (indicated by #). When p < 0.05:

; when p

< 0.01: **, ##; when p < 0.001: ***, ###; and when p < 0.0001: ****, ####. BCO: Bacillus coagulons, BT: Bifidobacterium thermophilum, CB: Clostridium butyricum, EF: Enterococcus faecium, BAL: Bifidobacterium animalis lactis, BCU; Bifidobacterium crudilactis, BMC: Bifidobacterium mongoliense, LP; Lactobacillus plantarum.

Figure 3 shows a time series indicating the accumulation of gas production in a batch in vitro fermentation (in vitro static fermentation) of a prebiotic alone or in combination with a probiotic (symbiotic). The in vitro batch fermentation of the symbiotic combination is carried out with 1.00E+07 CFU/ml of probiotic and 0.1 g of prebiotic (same quantity in the condition where the prebiotic is present alone), in the presence of an inoculum of 3 % in faeces. The results indicate the mean (ml/g of substrate) +/- the standard deviation of 3 experimental replicates. For prebiotics, GF: GOS/FOS, Inu: inulin, 2FL: 2'FL, BG: beta-glucan, resistant starch: RS. For probiotics, BCO: Bacillus coagulons, BT: Bifidobacferium thermophilum, CB; Clostridium butyricum, EF; Enterococcus faecium, BAL; Bifidobacferium animalis lactis, BCU; Bifidobacferium crudilactis, BMC; Bifidobacferium mongoliense, LP; Lactobacillus plantarum.

Figure 4 shows the relative abundance of groups of bacteria beneficial for intestinal eubiosis at 24 hours after the start of the batch in vitro fermentation (in vitro static fermentation) in the presence of the prebiotic alone or in the presence of symbiotic combinations (prebiotic + probiotic). Beneficial bacteria groups selected include bifidobacteria, lactobacilli, Clostridium cluster IV, Clostridium cluster XlVa and the butyryl-CoA:acetyl-CoA transferase gene. The qPCR 2 ^{^} (-ΔΔCt) measurement method was used to establish the relative abundance, the measurements were normalized to the total bacteria and a mixture of DNA samples from all extracts was used as a calibrator. The in vitro fermentation, in an individual fermenter, of the symbiotic combinations was carried out at L00E+07 CFU/ml of probiotic and 0.1 g of prebiotic (same quantity for the condition with the prebiotic alone), in the presence of 3% of a faeces inoculate. The results indicate the mean (ml/g of substrate) +/- the standard deviation of 3 experimental replicates. The statistical significance was analyzed by a one-way ANOVA test with a Dunnett type multiple comparison and where * corresponds to p < 0.05, ** to p < 0.01, *** to p < 0.01, and **** to p < 0.0001. For prebiotics, GF: GOS/FOS, Inu: inulin, 2FL: 2' FL, BG: beta-glucan, resistant starch: RS. For probiotics, BCO: Bacillus coagulons, BT: Bifidobacterium thermophilum, CB: Clostridium butyricum, EF: Enterococcus faecium, BAL: Bifidobacterium animalis lactis, BCU: Bifidobacterium crudilactis, BMO: Bifidobacterium mongoliense, LP; Lactobacillus plantarum.

Figure 5 shows the ability of the probiotic to survive and establish in a complex microbial community. Figure 5 shows the relative abundance of each probiotic compared to the total bacteria at 12 h, 24 h and 48 h after the start of fermentation for different symbiotic compositions. The relative abundance of each probiotic is normalized to the relative amount of probiotic obtained in the sample where only the probiotic was added. For prebiotics, GoF: GOS/FOS, Inu: inulin, 2FL: 2' FL, BG: beta-glucan, resistant starch: RS. For probiotics, BCO: Bacillus coagulons, BT: Bifidobacferium thermophilum, CB; Clostridium butyricum, EF; Enterococcus faecium, BAL; Bifidobacferium animalis lactis, LP; Lactobacillus plantarum.

FIG. 6 shows the evolution of the bacterial population between the end of the stabilization period and the end of the week of treatment: The results are shown with the following symbiotic compositions Enterococcus faecium (EF)+beta-glucan (BG); Bifidobacferium thermophilum (BT) + inulin (Inu); Clostridium butyricum (CB) + GOS/FOS (G/F); Bifidobacferium animalis lactis (BAL) + 2' FL (2FL); Bifidobacferium crudilactis (BG) + beta-glucan (BG); and the control condition.

The results correspond to the relative abundances of the quantity of DNA of the target bacterium relative to the quantity of DNA of the total bacteria (method of measurement by qPCR 2 ^{^} (-ΔΔCt)). FIG. 7 shows the impact of a symbiotic composition comprising Bifidobacterium thermophilum and inulin on the production of volatile fatty acids (VFA), SCFA, between the end of the stabilization period and the end of the week of treatment.

FIG. 8 shows the impact of a symbiotic composition comprising Bacillus animalis lactis and 2′ FL on the production of volatile fatty acids (VFA), SCFA, between the end of the stabilization period and the end of the week of treatment.

FIG. 9 shows the impact of a symbiotic composition comprising Bacillus crudilactis and beta-glucan on the production of volatile fatty acids (VFA), SCFA, between the end of the stabilization period and the end of the week of treatment.

FIG. 10 shows the impact of a symbiotic composition comprising Enterococcus faecium and beta-glucan on the production of volatile fatty acids (VFA), SCFA between the end of the stabilization period and the end of the week of treatment.

As indicated above, the present invention relates to a symbiotic composition comprising a therapeutically or preventively effective amount of (i) Enterococcus faecium, or (ii) Bifidobacterium animalis lactis, or (iii) Clostridium butyricum, or (iv) Bifidobacterium crudilactis, as a probiotic, and one or more prebiotics.

Preferably chosen from the group consisting of inulin, beta-glucan, 2' Fucosyllactose, GOS/FOS, resistant starch and a mixture thereof.

It has in fact been demonstrated according to the present invention that the symbiotic formulation containing a probiotic and a prebiotic produces a reduction in the production of IL-8 in an in vitro test, and a stabilization or an increase in “Short Chain Fatty Acids, SCFAs » as well as an improvement in the diversity and quantity of endogenous bacterial populations such as Lactobacillus, bifidobacteria, Clostridium clusters IV and XlVa as well as bacteria involved in the metabolic pathways of butyrate production, forming a balanced intestinal microbiota, leading in this way to generation of an anti-inflammatory environment in the digestive system of suckling and/or post-weaning piglets.

Administered to the pregnant sow and/or to the piglet, the symbiotic formulation according to the present invention promotes the establishment of a balanced intestinal microbiota very early, protecting the pregnant animal or even the newborn, more particularly the pregnant sow or piglet sustainably against dysbiosis with the help of the probiotic and prebiotic it contains, which together generate an anti-inflammatory environment.

Preferably, the combination is chosen from the group consisting of Clostridium butyricum and inulin, Clostridium butyricum and GOS/FOS, Bifidobacterium animalis lactis and inulin, Bifidobacterium animalis lactis and 2'fucosyllactose, Enterococcus faecium and beta-glucans, and Bifidobacterium crudilactis and beta- glucans, so as to allow a daily dosage of probiotic of between 1.00E+05 and 1.00E+015 CFU/day/animal, preferably between 1.00E+06 and 1.00E+013 CFU/day/animal, of advantageously between 1.00E+07 and 1.00E+011 CFU/day/animal, and a daily prebiotic dosage of between 0.1 and 1000 g/day/animal, preferably between 0.5 and 100 g/day/ animal, advantageously between 1 and 25 g/day/animal.

Advantageously, according to the invention, the probiotic consists for at least 80% by weight of a single probiotic chosen from the group consisting of (i) Enterococcus faecium, (ii) Bifidobacterium animalis lactis, (iii) Clostridium butyricum, and ( (iv) Bifidobacterium crudilactis.

Preferably, according to the invention, the composition comprises a single probiotic chosen from the group consisting of (i) Enterococcus faecium, (ii) Bifidobacterium animalis lactis, (iii) Clostridium butyricum, and (iv) Bifidobacterium crudilactis.

Advantageously, the therapeutically or preventively effective amount of probiotic is between 1.00E+05 and L00E+015 CFU/day/animal, preferably between 1.00E+06 and L00E+013 CFU/day/animal, advantageously between 1.00E+07 and 1.00E+01 1 CFU/day/animal. Preferably, the amount of said one or more prebiotics is between 0.1 and 1000 g/day/animal, preferably between 0.5 and 100 g/day/animal, advantageously between 1 and 25 g/day/animal.

Static fermentation in vitro:

The fermentation capacity of the different symbiotic combinations was tested in a static in vitro fermentation model, following the protocol described by Bindelle et al. Journal of Animal Science 2010, 87, 583-93. https://doi.orq/10.2527/ias.2007-0717 and by Tran et al. FEMS Microbiology Ecology 2016, 92, 1-13. https://doi.org/10.1093/femsec/fiyl 65. In summary, in vitro fermentation was carried out in flasks containing 15 ml of a solution containing 3% pre-weaning piglet faeces; three mucin-coated media previously immersed in mucin agar; 100 mg of prebiotic and 1.00E+07 CFU / ml of probiotic. Sealed vials were incubated at 39°C with shaking for 48 h in a water bath. Three replicates were performed per test condition.

Gas production:

The gas pressure was measured 2, 5, 8, 12, 16, 20, 24 and 48 h after the start of fermentation to determine the kinetics of fermentation using the monophasic mathematical model of Groot et al. Animal Feed Science and Technology 1996, 64, 77-89. https://doi.org/10.1016/S0377-8401(96)01012-7. Several parameters were calculated with this model: A (maximum gas volume, ml/g substrate), B (time to reach A/2, h), C as constant determining the slope of the inflection point of the profile, Rmax (rate fermentation, ml/g of substrate/hour) and Tmax (time to reach Rmax (h)).

Short chain fatty acids:

Lactate and short chain fatty acids (SCFA) were measured in the fermentation juice obtained after 24 hours by isocratic HPLC. The standard curve used contained acetate, propionate, butyrate as well as branched chain fatty acids (BCFA): isobutyrate, valerate and isovalerate.

The SCFA measurement was then calculated taking into account the basal production in the control-mucin flasks containing the microcosm carriers with mucin. The values are given in mmol per g of substrate and compared to measurements in the fermentation juice containing only the corresponding prebiotic.

Determination of microbiota modifications by qPCR during an in vitro static fermentation model test in the presence of different symbiotic compositions.

For each test in the in vitro static fermentation system, the analysis of changes in the intestinal microbiota was carried out by qPCR on samples taken after 24 hours of fermentation. The sequences of the forward primers and the antisense primers used to measure the expression by qPCR of the target genes corresponding to the bacterial populations beneficial to the microbial community of the intestine are indicated in Table 1 below.

Table 1.- Sequences of sense (F) and antisense (R) primers to measure by qPCR target genes corresponding to bacterial populations beneficial to the microbial community of the intestine.

Total bacteria are targeted using the primers described in Amit-Romach E. et al. Poult Sci 2004, 83, 1093-1098, doi: 10.3382/ps/pey394.

Bacteria belonging to the lactobacillus group are targeted using the primers described in Wang R. F. et al. Appl Environ Microbiol 1996, 62, 1242-1247, doi: 10.1 128/AEM.62.4.1242-1247.1996. The bacteria belonging to the bifidobacterium group are targeted using the primers described in Langendijk P. S. et al. Appl Environ Microbiol 1995, 61, 3069-3075, doi:10.1128/AEM.61.8.3069-3075.1995.

The bacteria forming part of the clostridium cluster IV group are targeted using the primers described in Matsuki T. et al. Appl Environ Microbiol 2004, 70, 7220-7228, doi: 10.1 128/AEM.70.12.7220-7228.2004. The bacteria forming part of the clostridium cluster XlVa group are targeted using the primers described in Matsuki T. et al. Appl Environ Microbiol 2002, 68, 5445-5451, doi:10.1128/aem.68.11.5445-5451.2002.

Butyril-CoA:acetate-CoA transferase is targeted using the primers described in Uerlings J. et al. J Sci Food Agric 2019, 99, 5720-5733, doi: 10.1002/isfa.9837.

Determination of the ability of probiotics to survive and establish themselves in a complex microbial ecosystem by qPCR during an in vitro static fermentation model assay in the presence of different symbiotic compositions.

For each test in the in vitro static fermentation model, the analysis and monitoring of the presence of each probiotic in the fermentation juice was carried out by qPCR on samples taken 12, 24 and 48 hours after the start of fermentation. . The sequences of the forward primers and the antisense primers used to measure the relative abundance of the probiotics by RT-qPCR are indicated in Table 2 below.

Table 2.- Sequences of forward (F) and antisense (R) primers for measuring the relative abundance of probiotics by qPCR.

References in Table 2: Veiga, P., et al. 2010. Proceedings of the National Academy of Sciences of the United States of America 2010, 107: 18132-37, https://doi.org/10.1073/pnas.101 1737107.

Hidaka T., et al. Wafer Research 2010, 44, 2554-62, https://doi.org/10.1016/j.watres.2010.01.007. Mathys S. et al. BMC Microbiology 2008, 8, 179, https://doi.org/10.1186/1471-2180-8-179.

Cremonesi P. et al. Journal of Dairy Research 2012, 79,318-23, https://doi.org/! 0.1017/S002202991200026X.

Loquasto JR et al. Applied and Environmental Microbiology 2013, 79, 6903-10, https://doi.org/! 0.1 128/AEM.01777-13.

Hoormon M. et al. Applied and Environmental Microbiology 2005, 71, 2318-24, https://doi.org/! 0.1128/AEM.71.5.2318.

Determination of the immune response caused by incubation of the fermentation juice obtained during an in vitro static fermentation model test in the presence of different symbiotic compositions.

In the context of the present invention, the IPEC-J2 line (intestinal enterocytes isolated from the jejunum of a newly born and non-breastfed piglet (or fed with a milk formulation) and belonging to a cell line neither transformed nor tumorigenic) was incubated in the fermentation juice produced by the symbiotic combination to be tested or containing only the prebiotic in question for 24 hours. Following this incubation period, the immune response is evaluated by the production of induced cytokines (IL-8, IL-6, IL-1 b) and TNF-alpha.

Previously, the cytotoxicity of the fermentation juices on the IPEC-J2 cells was evaluated because, in order to be able to observe an immune response, it is important that the cell line remains viable. For each combination selected, the effect of the different concentrations of fermentation juice, previously filtered at 0.8 μm, on the viability of the IPEC-J2 cells was measured by the MTT test (MTT tetrazolium salt: 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium), which is a colorimetric test for counting viable cells. Contact with the IPEC-J2 cells was then carried out with a fermentation juice inoculation rate of 0.8% (V/V).

Measurement of IL-8 concentration:

In the context of the present invention, the concentration of IL-8 was measured using in vitro tests on IPEC-J2 cells in the presence of the fermentation juice (containing or not containing the symbiotic composition to be tested) (see Figure 2 ) . To carry out the test, a test was carried out comparing the concentrations of IL-8 (the other markers IL-6, IL-1 b and TNF-alpha were not detected) produced in the presence of the fermentation juice in different conditions: control-mucin (horizontal line in Figure 2), prebiotics alone (GOS/FOS (GF), 2FL, Inulin (Inu), resistant starch (RS) and (3-glucan (BG)) conditions, and symbiotic ( other conditions in Figure 2. This assay indicates that if the level of IL-8 is lower in the presence of the fermentation juice containing the prebiotic or symbiotic composition compared to the control fermentation juice-mucin, it shows an anti-inflammatory effect. the presence of the pre- or symbiotic In addition, if the level of IL-8 is lower in the presence of the fermentation juice containing the symbiotic composition compared to the fermentation juice containing only the prebiotic, this shows that the specific combination of said au at least one probiotic with said at least one prebiotic has an additional anti-inflammatory effect compared to the prebiotic alone. The cell viability of the synbiotics (results of the MTT test) was taken into account to adjust the concentrations of IL-8. The level of expression of IL-8 is determined by ELISA (Porcine IL-8/CXCL8 DuoSet ELISA kit DY535: R&D Systems).

Baby SPIME model:

The BabySPIME model is a scientifically validated model (Dufourny S. et al. Journal of Microbiological Methods, 2019, 167: 105735. https://doi.Org/10.1016/j.mimet.2019) that simulates physiological dynamics and conditions of a complete gastrointestinal tract in piglets in an in vitro environment (equipment derived from the SHIME model of ProDigest Bvba, Gent, Belgium). The model includes 2 times 3 reactors that sequentially simulate the stomach (acid condition and digestion by pepsin), the ileum (process of enzymatic digestion) and the proximal colon (process of fermentation by microbiota) (see Figure 1 ). This system makes it possible to obtain complex and stable microbial communities whose function and structure are strongly similar to the microbial communities found in the different regions of the intestine.

Peristaltic pumps allow the transfer of culture medium, pancreatic juice, bile, acid (0.5M HCl), base (0.5M NaOH) and fermentation liquids from one reactor to another during a complete cycle. In practice, reactor 1 simulates the functions of the stomach, duodenum and jejunum, reactor 2 simulates the functions of the ileum, reactor 3 simulates the functions of the proximal colon.

A full trial in the babySPIME system lasts 3 weeks: 2 weeks of microbiota stabilization followed by a week of treatment with the symbiotic composition. A solution of 1.00E+08 CFU/ml was prepared to inoculate a final dose of 1.00E+07 CFU/ml of intestinal contents. The dose of prebiotic is 2 g of prebiotic/day to reach a concentration of 1% of the diet.

The faeces of 6 suckling 27-day-old piglets that had not undergone antibiotic treatment were used to prepare the inoculum for the study. Faeces were collected directly from the piglets and kept on ice under anaerobic conditions. For each test, the inoculum was then homogenized for 10 minutes by adding the faeces of each piglet to a phosphate buffer solution under anaerobic conditions. After macroscopic filtration to eliminate the particles still in suspension, the filtrate was injected simultaneously into reactors 2 and 3. Before inoculation, these two reactors were filled with a non-acidified culture medium and the pH was automatically adjusted in each reactor so as to best simulate the physiological conditions.

The culture medium was prepared and validated beforehand. Bottles of medium were stored at 4°C and the pH was adjusted to 3.0 before use in the first reactor. A solution mimicking pancreatic juice containing sodium bicarbonate (2.5 g/L, VWR Chemicals, Radnol, Pennsylvania, USA), pancreatin (0.9g/L, ProDigest) and bile salts (Oxgall 4.0 g/L) a was added in the middle.

The feeding cycle is scheduled 3 times per day based on a total retention time of 14h. During each cycle, the culture medium, maintained at 4° C., passes through reactor 1 for 1 hour 30 minutes. Then, the mixture of pancreatic juice/bile salts (60ml), also maintained at 4° C., is added to the same reactor for 1 hour. After this time, and simultaneously, the contents of reactors 1, 2 and 3 are transferred, respectively, into reactors 2, 3 as well as a biological waste container.

The anaerobic conditions of all the reactors are maintained thanks to a flow of nitrogen (N2) once a day for 10 minutes at the level of the reactors. The contents of the reactors are stirred continuously (300 rpm) and maintained at 39.5°C. The pH of reactors 2 and 3 is continuously monitored in order to stabilize the pH of the ileum and the proximal colon.

Samples were collected in the babySPIME system during the stabilization phase, before the treatment with the symbiotic formulations and at the end of the treatment with the symbiotic.

For each trial in the babySPIME system, the analysis of the changes in the intestinal microbiota was carried out by qPCR on samples taken at the end of the stabilization phase and at the end of the week of treatment with a symbiotic composition. The sequences of the sense primers and of the anti-sense primers used to measure by qPCR the relative quantity of the target genes corresponding to the bacterial populations beneficial for the microbial community of the intestine are indicated in Table 1 mentioned above.

Three repetitions on the BABY-SPIME model were carried out for each of the symbiotic combinations.

Examples.-

Example 1: The fermentation capacity of the symbiotic composition by modeling the production of gas and the production of lactate and SCFA in an in vitro static fermentation model

The fermentation curves of the pre- or symbiotics are shown in Figure 3. Under all conditions in Figure 3, there is an increase in fermentation capacity measured by the increase in gas production. Under all the conditions of FIG. 3, this gas production is greater in the presence of a symbiotic composition compared to the presence of the prebiotic alone. The parameters resulting from the modeling of the gas production during the static fermentation of the symbiotics are compared with the parameters of the prebiotics alone (Table 3).

The gas production kinetics and model parameters include: A: maximum gas production; B: the time to have 50% of the maximum gas production; C: the slope; D: the maximum rate of gas production (Rmax); the time at Rmax (Tmax). The static fermentation of the symbiotic combinations was carried out at 1.00E+7 CFU/ml of probiotic and 0.1 g of prebiotic (same quantity of prebiotic used in the condition where the prebiotic is alone) in the presence of 3% of an inoculum of faeces. Results show the mean +/- standard deviation of 3 experimental replicates. The p-values were obtained by a one-way ANOVA statistical analysis with a Dunnett type multiple comparison test. Results are considered significant when p < 0.05.

The SCFAs present in the fermentation juice after 24 h of fermentation are presented in Table 4. There was no production of lactate after 24 h of fermentation.

The static fermentation of the symbiotic combinations was carried out at 1.00E+07 CFU/ml of probiotic and 0.1 g of prebiotic (same quantity of prebiotic used in the condition where the prebiotic is alone) in the presence of 3% of a faeces inoculate. Results show the mean +/- standard deviation of 3 experimental replicates. The p-values were obtained by a one-way ANOVA statistical analysis with a Dunnett type multiple comparison test. When p < 0.05: *; p < 0.01: **; p < 0.001: ***; p < 0.0001: ****.

Example 2: Determination of changes in the microbiota by qPCR during an in vitro static fermentation model test in the presence of different symbiotic compositions

The relative abundance of beneficial bacteria after 24 h of fermentation are presented in Figure 4. The delta-delta CT method was used to express the target bacteria compared to the total bacteria. The results of the synbiotics are compared with the prebiotics alone. As shown in Figure 4, several symbiotic compositions, such as BT-GF, CB-GF, BCO-GF, BCU-2FL, BMO-2FL, BAL-2FL, BAL-Inu, BT-Inu, CB-RS, BCU -RS, BAL-BG, LP-BG, or BCU-BG, make it possible to significantly increase the relative abundance of beneficial bacteria at 24 hours of fermentation.

Example 3: Determination of the ability of probiotics to survive and establish during fermentation

Figure 5 shows the presence of probiotics in the fermentation juice after 12, 24 and 48 hours of fermentation. Symbiotic compositions, such as BT-Inu, allow good establishment and survivability of the probiotic.

Example 4: Determination of the immunomodulatory effect of the symbiotic composition by quantification of the production of IL-8 and NO by IPEC-J2 cells.

IL-8 production in IPEC-J2 cells was measured by an assay

ELISA.

After 24 h of incubation, the immune response was evaluated by measuring the production of IL-8 in the cell culture medium. The IPEC-J2 cells were incubated with 0.8% (V/V) of fermentation juice previously sterilized by filtration. Moreover, the survival rate of IPEC-J2 cells is greater than or equal to 70% in all the tested conditions of fermentation juice.

To determine the impact of the symbiotic composition contained in the fermentation juice on the production of IL-8, the production rate of IL-8 was measured in a control condition where the fermentation juice contains a faeces inoculum. , mucin (control-mucin), but in the absence of probiotics, prebiotics or symbiotics. In this control condition, a basal level of IL-8 expression is 932 pg/ml (represented by the horizontal line in Figure 2). Pre- and synbiotics were compared against control-mucin. In addition, the synbiotics were compared with the prebiotic to estimate the additional effect of the probiotic. Table 5 shows the p-values of these comparisons, and Figure 2 shows the concentrations of IL8.

Table 5.- Production of IL-8 by the IPEC-J2 cells in the supernatant after 24 hours of incubation with 0.8% of fermentation juice previously sterilized by filtration produced by the symbiotic composition or the prebiotic alone.

Values are corrected for cell viability. The results are means with their standard deviations. The comparisons are made between the symbiotic compositions and the prebiotic alone. The statistical significances are analyzed by a one-way ANOVA with a Dunnett type multiple comparison test. When p<0.05: *; p<0.01: **; p<0.001: ***; p<0.0001: ****.

Example 5: Determination of the modifications of the microbiota by qPCR during a test in the baby SPIME model in the presence of different symbiotic compositions. The overall results obtained on the baby SPIME system are presented in Figure 6 and Table 6.

Table 6.- Summary of the upward or downward regulation of bacterial groups of interest following symbiotic treatment in the BABY-SPIME system after one week of treatment with the various symbiotics.

We find either a decrease in the bacterial population, an increase in the bacterial population or a stabilization of the bacterial population. The statistical test performed is a Wilcoxon test. Example 6: Symbiotic composition C1 comprising at least

Clostridium butyricum and at least GOS/FOS against intestinal dysbiosis.

Composition C1 was prepared by adding 1 E07 CFU/ml Clostridium bufyricum and 0.1 g GOS/FOS.

More specifically, at the prebiotic level, 0.1 g of GOS/FOS was added to each fermentation bottle containing 15ml of fermentation juice. For the GOS/FOS mixture, the prebiotic consists of a homogeneous liquid solution of 80% (w/w) GOS and 20% (w/w) FOS. The level of probiotic, a freeze-dried powder of bacteria was pre-mixed in a buffer solution at a concentration of 1.5E08 CFU/ml (stock solution). Before being added to the fermentation flask, the pre-mixed probiotic solution underwent a revivification process consisting of a 30 min incubation at 37°C with shaking. Then, 1 ml of the pre-mixed solution (stock solution) was added to the vial leading to a final probiotic concentration of 1 E07 CFU/ml in the vial.

The symbiotic composition comprising at least Clostridium butyricum and at least GOS/FOS against intestinal dysbiosis showed a favorable effect on fermentation, by increasing the A value (maximum gas production) compared to the prebiotic alone (see CB-GoF condition of the Table 3, and by increasing the concentrations of total SCFAs and butyrate after 24 hours of static fermentation in vitro (see CB-GoF condition of Table 4) In addition, the relative abundance of Bifidobacteria and lactobacilli increased in comparison with the prebiotic alone (see CB-GF condition in Figure 4).The probiotic Clostridium butyricum was still present in the fermentation juice after 48 hours of fermentation.An anti-inflammatory effect is shown in vitro on IPEC-J2 cells generating a decrease in the IL-8 production compared to GOS/FOS alone, while IL-8 concentration was not different compared to control-mucin (see CB-GF condition in Figure 2).

Moreover, the Cl symbiotic composition shows an improvement of the microbiota in the BABY-SPIME model by increasing the bacterial populations involved in the metabolic pathways of butyrate production. The increase in these bacterial populations was analyzed by qPCR (see condition CB+G/F of FIG. 6 and Table 6 presented above). The sequences of the forward primers and the antisense primers used to measure by qPCR the relative abundance of the beneficial bacterial populations for the microbial community of the intestine are indicated in Table 1.

Example 7: C2 symbiotic composition comprising at least Bifidobacterium thermophilum and at least inulin against intestinal dysbiosis.

Composition C2 was prepared by adding 1 E07 CFU/ml of Bifidobacterium thermophilum and 0.1 g of inulin.

More specifically, at the level of the prebiotic, 0.1 g of inulin was added to each fermentation bottle containing 15 ml of fermentation juice. With regard to the probiotic Bifidobacterium thermophilum, a freeze-dried powder of bacteria was pre-mixed in a buffer solution at a concentration of 1.5E08 CFU/ml (stock solution). Before being added to the fermentation flask, the pre-mixed probiotic solution underwent a revivification process consisting of a 30 min incubation at 37°C with shaking. Then, 1 ml of the pre-mixed solution (stock solution) was added to the vial leading to a final probiotic concentration of 1 E07 CFU/ml in the vial.

The C2 symbiotic composition comprising Bifidobacterium thermophilum (BT) and inulin (Inu) against intestinal dysbiosis showed a beneficial effect on fermentation, demonstrated by an increase in A value (maximal gas production), a decrease in B value (the time to reach 50% of maximum gas production), an increase in fermentation rate (Rmax), and a decrease in time to reach the maximum amount of gas produced (Tmax) compared to inulin alone (see BT-Inu condition in Table 3). In addition, an increase in the concentration of butyrate and the relative abundance of bifidobacteria in the fermentation juice after 24 hours is demonstrated (see BT-Inu condition in Figure 4), Bifidobacterium thermophilum was indeed present until the end of the fermentation. An in vitro anti-inflammatory effect on IPEC-J2 cells generating a decrease in IL-8 production compared to inulin alone (no difference between the synbiotic and the control-mucin) according to the protocol described at l Example 1 (see BT-Inu condition in Figure 2).

In addition, the C2 symbiotic composition shows an improvement of the microbiota in the BABY-SPIME model by increasing the bacterial populations involved in the metabolic pathways of butyrate production and showing a tendency towards an increase in bifidobacteria. The increase in these bacterial populations was measured by qPCR (see BT + INU condition of Figure 6 and Table 6). The sequences of the forward primers and the antisense primers used to measure by qPCR the relative abundance of the beneficial bacterial populations for the microbial community of the intestine are indicated in Table 1. In parallel, an analysis of VFAs (Figure 7) was able to show a trend towards an increase in valerate in the ileum and ascending colon fermenters (p = 0.066) and a trend towards an increase in total VFAs in the ileum (p = 0.090). Example 8: C3 symbiotic composition comprising at least Bifidobacterium animalis lactis and at least 2' FL against intestinal dysbiosis.

Composition C3 was prepared by adding 1 E07 CFU/ml of Bifidobacterium animalis lactis and 0.1 g of 2' FL.

More particularly, at the level of the prebiotic, 0.1 g of 2' FL was added to each fermentation flask containing 15 ml of fermentation juice. With regard to the probiotic Bifidobacterium animalis lactis, a freeze-dried powder of bacteria was pre-mixed in a buffer solution at a concentration of 1.5E08 CFU/ml (stock solution). Before being added to the fermentation flask, the pre-mixed probiotic solution underwent a revivification process consisting of a 30 min incubation at 37°C with shaking. Then, 1 ml of the pre-mixed solution (stock solution) was added to the vial leading to a final probiotic concentration of 1 E07 CFU/ml in the vial.

The C3 symbiotic composition against intestinal dysbiosis showed a beneficial effect on fermentation, demonstrated by an increase in the A value (maximum gas production), a decrease in the B value (the time to reach 50% of the maximum production of gas), an increase in the fermentation rate (Rmax), and a decrease in the time to reach the maximum quantity of gas produced (Tmax) compared to 2' FL alone (see Bal-2FL condition of Table 3). In addition, an increase in the butyrate concentration (see BAL-2FL condition in Table 4) and the relative abundance of Clostridium clusters IV in the fermentation juice after 24 hours is demonstrated (see BAL-2FL condition in Figure 4), Bifidobacterium animalis lactis was present until the end of fermentation. An in vitro anti-inflammatory effect on IPEC-J2 cells generating a decrease in the production of IL-8 (see BAL-2FL condition in Figure 2) compared to 2' FL alone and compared to the control-mucin according to the protocol of Example 1.

Moreover, the C3 symbiotic composition shows an improvement of the microbiota in the BABY-SPIME model by increasing the bacterial populations of bifidobacteria. The increase in these bacterial populations was measured by qPCR (see BAL + 2FL condition of Figure 6 and Table 6). The sequences of sense primers and antisense primers used to measure by qPCR the relative abundance of bacterial populations beneficial to the microbial community of the intestine are shown in Table 1. In parallel, an analysis of volatile fatty acids (VFA) (Figure 8) was able to show a significant increase in acetate in the ileum and ascending colon fermenters (p = 0.034) and in the fermenter mimicking the ileum (p = 0.023 ). A tendency for acetate to increase in the fermenter mimicking the ascending colon (p = 0.075) and a tendency for isobutyrate to decrease in the ileum and ascending colon fermenters (p = 0.076) and ascending colon (p = 0.095) were also observed.

Example 9: C4 symbiotic composition comprising Bifidobacterium crudilactis and beta-glucan against intestinal dysbiosis.

Composition C4 was prepared by adding 1 E07 CFU/ml of Bifidobacterium crudilactis and 0.1 g of beta-glucan.

More specifically, at the level of the prebiotic, 0.1 g of beta-glucan was added to each fermentation bottle containing 15ml of fermentation juice. With regard to the probiotic Bifidobacterium animalis lactis, a freeze-dried powder of bacteria was pre-mixed in a buffer solution at a concentration of 1.5E08 CFU/ml (stock solution). Before being added to the fermentation flask, the pre-mixed probiotic solution underwent a revivification process consisting of a 30 min incubation at 37°C with shaking. Then, 1 ml of the pre-mixed solution (stock solution) was added to the vial leading to a final probiotic concentration of 1 E07 CFU/ml in the vial.

The C4 symbiotic composition comprising Bifidobacferium crudilactis and beta-glucan against intestinal dysbiosis showed a beneficial effect on fermentation, demonstrated by an increase in the A value (maximum gas production) and an increase in the rate of fermentation (Rmax) compared to beta-glucan alone (see BCU-BG condition in Table 3). In addition, after 24 hours of fermentation, an increase in the concentration of total SCFAs and butyrate (see BCU-BG condition of Table 4) and the relative abundance of Clostridium clusters XlVa in the fermentation juice is demonstrated (see BCU-BG condition). BG of Figure 4). An anti-inflammatory effect is demonstrated in vitro on IPEC-J2 cells generating a decrease in the production of IL-8 compared to beta-glucan alone and compared to the control-mucin according to the protocol described in example 1 (see BCU condition -BG of Figure 2). In addition, the C4 symbiotic composition shows an improvement of the microbiota in the BABY-SPIME model by increasing the bacterial populations involved in the metabolic pathways of butyrate production and showing a tendency towards an increase in bifidobacteria. The increase in these bacterial populations was measured by qPCR (see BG + BG condition of Figure 6 and Table 6). The sequences of the forward primers and the antisense primers used to measure by qPCR the relative abundance of the beneficial bacterial populations for the microbial community of the intestine are indicated in Table 1.

In parallel, an analysis of VFAs (Figure 9) showed a significant decrease in isobutyrate (p = 0.018) in the ileal fermenters and ascending colons while a downward trend in total VFAs was observed in the ascending colon. (p = 0.058).

Example 10: C5 symbiotic composition comprising Enterococcus faecium and beta-glucan against intestinal dysbiosis.

The C5 symbiotic composition against intestinal dysbiosis showed a beneficial effect on fermentation, demonstrated by an increase in the A value (maximal gas production) and an increase in the rate of fermentation (Rmax) compared to beta-glucan alone ( see condition EF-BG in Table 3). In addition, after 24 hours of fermentation, an increase in the concentration of total SCFAs and butyrate in the fermentation juice is demonstrated (see EF-BG condition in Table 4).

In addition, the C5 symbiotic composition shows an improvement of the microbiota in the BABY-SPIME model by increasing the bacterial populations involved in the metabolic pathways of butyrate production. The increase in these bacterial populations was measured by qPCR (see EF + BG condition of Figure 6 and Table 6). The sequences of the forward primers and the antisense primers used to measure by qPCR the relative abundance of the beneficial bacterial populations for the microbial community of the intestine are indicated in Table 1.

In parallel, an analysis of VFAs (Figure 10) showed a significant decrease in valerate in the fermenter mimicking the ileum (p = 0.0096). A A decreasing trend in isovalerate was also observed in the ileum (p = 0.099) and in the ileum and ascending colon fermenters (p = 0.052).

Example 11: Symbiotic compositions administered in vivo to the pregnant and/or lactating sow and/or to the suckling piglet Table 7 below describes the various symbiotic compositions (SYN) given in vivo to the pregnant and/or lactating sow and/or nursing piglets.

Table 7.- Different symbiotic compositions (SYN) tested in vivo.

Example 12: In vivo supplementation protocol for sows and piglets.

Symbiotic supplementation of the sows started from the ^80th day of gestation and it was distributed manually in the form of a portion directly added to the plate of each sow (top feeding) once per day with a meal. Supplementation of the synbiotic continued throughout the three-week lactation (nursing) period. The synbiotics were prepared, ie mixed with a growth standard food, just before the start of the experiments and kept in a cool, dry place. A group of non-supplemented sows was used as a reference/control group for the comparison. The sows were distributed into different groups according to their parity, in order to obtain a homogeneous distribution and an equal average parity between the groups (4). After 80 days of gestation, the sows were placed in individual pens with individual feed automatically distributed twice a day. After 115 days of gestation, the sows were transferred to individual farrowing pens. The sows were fed twice a day with an automatic feeder and had unlimited access to water by pressing a button above their pan. The supplementation of the symbiotic compositions (examples 13, 15, 17, 19, 21 of table 7) was carried out once a day in the morning.

Table 8 below indicates the dose of probiotic and prebiotic administered to sows according to the stage of development (gestation or lactating).

Table 8.- Doses of symbiotic compositions distributed to sows.

The symbiotic combinations selected consisted of a single probiotic strain with a single prebiotic (see examples 13 (SYN 1), 15 (SYN 2), 17 (SYN 3), 19 (SYN 4) and 21 (SYN 5) in Table 7 ) . Symbiotic combinations that included a milk oligosaccharide as a prebiotic were replaced by inulin for sow supplementation. Examples 14 (SYN 1), 16 (SYN 2), 18 (SYN 3), 20 (SYN 4) and 22 (SYN 5) of Table 7 indicate the symbiotic compositions administered to suckling piglets.

During the suckling phase, the piglets remained with their mother in maternity pens whose floor was covered with a plastic mesh. The piglets had unrestricted access to suckling, and from the first day after birth a floor-mounted pan was placed in each pen. During the first week after birth, the piglets received a milk replacer once a day. During the second and third weeks of lactation, the piglets received a wet transition food.

The symbiotic supplementation of the piglets began the day after birth, each farrowing pen received a supplement of lacto-replacer in a plate fixed to the floor. Supplementation of the synbiotic continued throughout the three-week lactation period. Symbiotic supplementation was discontinued at weaning. Thus, no symbiotic supplementation was given to the piglets during the post-weaning period. Litters/piglets that received the synbiotics were compared to litters/piglets not supplemented.

The prebiotic solutions (dissolved in drinking water) were prepared once a week and stored in the refrigerator (+4°C). The probiotic powder and prebiotic solution were freshly mixed each day just before distribution. The symbiotic compositions were administered at a dose established for an average litter of 15 piglets receiving 2 ml of symbiotic per piglet per day during the first week, and 4 ml per piglet per day from the second week until weaning. The symbiotic composition was distributed to the piglets by administering it in the milk replacer given to them during the first week and with the liquid wet transition food until weaning.

Table 9 below indicates the dose of probiotic and prebiotic administered to piglets according to the stage of development.

Table 9.- Doses of symbiotic compositions distributed to piglets.

Examples 13 to 22: Different symbiotic compositions administered in vivo to sows and piglets.

Examples 13 (SYN 1), 15 (SYN 2), 17 (SYN 3), 19 (SYN 4) and 21 (SYN 5) of Table 7 indicate the symbiotic compositions administered to pregnant or lactating sows. Examples 14 (SYN 1), 16 (SYN 2), 18 (SYN 3), 20 (SYN 4) and 22 (SYN 5) of Table 7 indicate the symbiotic compositions administered to suckling piglets.

Examples 23 to 43: Impact of different administration protocols of a symbiotic composition to the sow and/or to the suckling piglet on different zootechnical criteria of the suckling piglet and/or post-weaning.

At birth, piglets were counted (live, dead and mummified) and the total litter weight of live piglets was recorded. One day before weaning, the piglets were counted and the total litter weight of live piglets was recorded. The piglets were weaned three weeks after farrowing, at around 21 days of age. At weaning, piglets were transferred to post-weaning pens in groups per treatment, rearranged by sex (male or female) and size (small, medium, large) into groups of approximately 25 piglets per pen. Two weeks after weaning, the piglets were counted and weighed.

Table 10 below indicates the different administration protocols of the symbiotic composition to the sow and/or to the suckling piglet.

Table 10.- Different protocols/profiles for administration of the symbiotic composition to the sow and/or to the piglet.

Table 7 indicates the prebiotic and probiotic composition of each SYN indicated in Table 10. Tables 8 and 9 show the doses administered. Different zootechnical criteria were measured on the suckling piglet and on the post-weaning piglet according to different administration profiles of the different symbiotic compositions. These different zootechnical criteria are: the weight of the piglet (kg) (see table 1 1 ) the average daily gain, GQM (g/day) (see table 12) the quantity (score) of diarrhea (%) (see table 13)

Diarrhea scores were measured by assigning each pen/litter the highest score according to the diarrhea score scale based on feces consistency. The rating scale used included five categories: score 0 - hard granule; 1 - soft and dry granule; 2 - wet granule/soft form; 3 - soft, unformed granule; 4 - watery. Only score 4 was considered diarrhea. Observations of diarrhea occurred twice during the experiment, 3 days before weaning and 10 days after weaning. The days of observation of the diarrhea scores were chosen not to coincide with the weighing of the piglets nor with the collection of faeces in order to avoid interfering with the results due to the stress caused by the handling on the piglets.

Table 11 below shows the impact of different SYN administration profiles on piglet weight during suckling and post-weaning.

Table 11.- Suckling and post-weaning piglet weight according to different SYN administration profiles.

Certain administration profiles of symbiotic compositions, such as SYN 1 and SYN 2 (example 26 or 29) (see Table 10), allow a significant increase in piglet weight compared to the control condition in the post-weaning phase or in the post-weaning phase. breastfeeding (see for example SYN 3 in Example 32). The symbiotic compositions involving SYN 5 (Examples 40 to 43) are counter-examples showing that certain symbiotic compositions will not necessarily be positive for the weight gain of the suckling piglet and/or post-weaning. Table 12 below shows the impact of different SYN administration profiles on the average daily gain (ADG) of suckling and post-weaning piglets.

Table 12.- GQM of suckling and post-weaning piglets according to different SYN administration profiles.

The results in Table 12 show that the administration profiles involving SYN 1 (example 26), SYN 3 (examples 32, 33) and SYN 4 (examples 34-36) allow a better average daily gain of the piglet compared to the condition control, in particular for post-weaning piglets.

Table 13 below shows the impact of different SYN administration profiles on the diarrhea score of suckling and post-weaning piglets.

Table 13.- Diarrhea in suckling and post-weaning piglets according to different SYN administration profiles.

The administration profiles involving SYN 3 and SYN 4 allow a reduction in severe diarrhea (score 4) in the piglet compared to a control piglet, in particular for the post-weaning piglet.

These results show that depending on the symbiotic composition and depending on the administration profile, certain compositions will have a direct positive impact (compared to the control condition) on the zootechnical criteria for suckling piglets while others will have a persistence effect by having a more marked positive impact on zootechnical criteria for post-weaning piglets.

Example 44 Synbiotic composition SYN 6 comprising E. faecium, C. butyricum, beta-glucan and inulin against intestinal dysbiosis and doses administered in vivo to the sow before farrowing and after farrowing.

For probiotics, the amount administered is indicated in CFU/sow/day, while for prebiotics the amount administered is indicated in g/sow/day (see Table 14).

The in vivo administration of the symbiotic composition was carried out at 4 different stages of development: before whelping, one week after whelping, 2 weeks after whelping, 3 weeks after whelping and 4 weeks after whelping.

On the 76th day of gestation, the sows were weighed. The sows were then divided into three different groups according to their parity, in order to obtain a homogeneous distribution and an equal average parity in all the groups. On day 80 after insemination, the sows were placed in individual pens. On the 107th day of gestation, the sows were weighed. On the 108th day of gestation, the sows were transferred to individual pens in the farrowing house. Parturition was planned for day 1 of gestation with an injection of Planate® and uterine involution was facilitated by an injection of dinolytic® after parturition.

At birth, the piglets were counted and their individual weights were recorded. The piglets were housed with the sows in maternity pens with continuous suckling access. From day one, after all sows had farrowed, litter size was balanced by adoptions between litters of the same treatment. The piglets were weighed individually every week. In this experiment, piglets were weaned four weeks after farrowing, at around 28 days of age. At weaning, a selection of 2 males and 2 females from the same litter was made based on the average body weight in the treatment. When this was not possible, piglets of appropriate weight and sex were chosen from another litter of the same treatment group. Selected piglets were transferred to post-weaning pens in groups per treatment.

During the post-weaning period, they received a diet devoid of any dietary supplement (devoid of any symbiotic composition). The piglets were checked daily in the morning to check their general state of health and monitor the appearance of diarrhoea. Feeders were filled as needed. The enclosures were cleaned with water every day. Once a week, the piglets were weighed and feed consumption was recorded.

Table 14.- SYN 6 symbiotic composition comprising E. faecium, C. butyricum, beta-glucan and inulin and quantity of each probiotic and prebiotic administered per day at 5 stages of development.

MB: farrowing; G: day of gestation

The 5 stages of development are G80-G107 (between 80 days and 107 days of gestation, before parturition), 1 week after parturition, 2 weeks after parturition,

3 weeks postpartum and 4 weeks postpartum. Example 45: SYN 7 symbiotic composition comprising E. faecium,

B. animalis lactis, B. crudilactis, beta-glucan and inulin against intestinal dysbiosis and doses administered in vivo to the sow before farrowing and after farrowing.

For probiotics, the amount administered is indicated in CFU/sow/day, while for prebiotics the amount administered is indicated in g/sow/day (see Table 15).

The in vivo administration of the symbiotic composition was carried out at

4 different stages of development: G80-G107 (between 80 and 107 days of gestation, before parturition, MB), one week after parturition, 2 weeks after parturition and 3 weeks after parturition. Table 15.- SYN 7 symbiotic composition comprising E. faecium,

B. animalis lactis, B. crudilactis, beta-glucan and inulin and quantity of each probiotic and prebiotic administered per day at 5 stages of development.

MB: farrowing; G: day of gestation

Example 46: Symbiotic composition against intestinal dysbiosis and doses administered in vivo to the piglet

In an in vivo test (first experiment), the symbiotic 1 (SYN 1) is composed of Clostridium butyricum and GOS/FOS (80/20) and the symbiotic 2 (SYN 2) is composed of B. animalis lactis and 2' FL (see Table 16). The control condition was just potable water. In a controlled manner, the piglets ingested a determined amount of symbiotic, which increased with age. The symbiotic composition was prepared daily by mixing a prebiotic mixture of 0.4 g of prebiotic per ml in drinking water with the corresponding probiotic (6E08 CFU/ml). The GOS/FOS prebiotic mixture contained 0.32 g/ml GOS and 0.08 g/ml FOS according to the proportions of sow milk oligosaccharides. The prebiotic solution was prepared weekly and stored at +4°C until use. Prior to administration, the symbiotic mixtures were brought to room temperature. The symbiotic compositions were administered from birth, at increasingly shorter intervals and at increasing doses with age, until weaning on the 28th postnatal day (see Table 16). No symbiotic composition was administered during the post-withdrawal period. The volumes of the symbiotic compositions administered were as follows:

1.25 ml/piglet up to and including the following Saturday,

1.75 ml/piglet the following two days,

2.5 ml/piglet until Friday inclusive of the following week,

3.75 ml/piglet the second week from Monday,

5 ml/piglet then from Monday of the third week. Table 16.- Volumes and doses of symbiotic compositions administered to piglets during the lactation phase from birth to weaning (D28).

It has been observed that the administration of a symbiotic composition comprising several probiotics, such as SYN 6 or SYN 7 (examples 44 and 45), does not allow greater weight gain in the piglet during lactation and/or post-weaning, nor a greater average daily gain of the suckling and/or post-weaning piglet compared to a composition comprising a single probiotic, such as SYN1 or SYN 2 (example 46), or compared to a control condition (control piglet) having no not received a symbiotic composition. In addition, the synbiotic compositions SYN 6 and SYN 7 increase the amount of diarrhea (score 4) in suckling piglets by 10% and 17% respectively compared to the control condition. It is understood that the present invention is in no way limited to the embodiments described above and that many modifications can be made thereto without departing from the scope of the appended claims.

Claims

"Claims"

1. Symbiotic composition for the treatment or prevention of dysbiosis in suckling and/or post-weaning piglets comprising a therapeutically or preventively effective amount of

(i) Enterococcus faecium, or

(ii) Bifidobacterium animalis lactis, or

(iii) Clostridium butyricum, or

(iv) Bifidobacterium crudilactis, as a probiotic, and one or more prebiotics, this composition being to be administered to a pregnant or lactating sow, and/or to a nursing and/or post-weaning piglet, to promote an anti -inflammatory in the intestine of said suckling and/or post-weaning piglet.

2. Composition according to claim 1, in which said one or more prebiotics is chosen from the group consisting of inulin, beta-glucan, 2' Fucosyllactose, GOS/FOS (galacto-oligosaccharide, fructo-oligosaccharide), resistant starch and mixtures thereof.

3. Composition according to claim 1 or claim 2, in which the therapeutically or preventively effective amount of probiotic is between 1.00E+05 and L00E+015 CFU/day/animal, preferably between 1.00E+06 and 1, 00E+013 CFU/day/animal, advantageously between 1.00E+07 and 1.00E+011 CFU/day/animal.

4. Composition according to any one of the preceding claims, in which an amount of said one or more prebiotics is between 0.1 and 1000 g/day/animal, preferably between 0.5 and 100 g/day/animal, so advantageous between 1 and 25 g/day/animal.

5. Composition according to any one of the preceding claims, in which the said probiotic consists for at least 80% by weight of a single probiotic chosen from the group consisting of (i) Enterococcus taecium, (ii) Bifidobacterium animalis lactis, (iii) Clostridium butyricum, and (iv) Bifidobacterium crudilactis.

6. Composition according to any one of the preceding claims, characterized in that it comprises a single probiotic chosen from the group consisting of (i) Enterococcus faecium, (ii) Bifidobacterium animalis lactis, (iii) Clostridium butyricum, and (iv ) Bifidobacterium crudilactis.

7. Composition according to any one of the preceding claims, in the form of a combination comprising said probiotic in a therapeutically or preventively effective amount and said one or more prebiotics, the combination being chosen from the group consisting of Bifidobacterium crudilactis and beta-glucans, Bifidobacterium crudilactis and inulin, Bifidobacterium crudilactis and 2'fucosyllactose, Bifidobacterium crudilactis and GOS/FOS, Bifidobacterium crudilactis and resistant starch, Bifidobacterium animalis lactis and 2'fucosyllactose, Bifidobacterium animalis lactis and inulin, Bifidobacterium animalis lactis and beta-glucans, Bifidobacterium animalis lactis and GOS/FOS, Bifidobacterium animalis lactis and resistant starch, Clostridium butyricum and GOS/FOS, Clostridium butyricum and inulin, Clostridium butyricum and beta-glucans, Clostridium butyricum and 2'fucosyllactose, Clostridium butyricum and resistant starch, Enterococcus taecium and beta-glucans, Enterococcus taecium and inulin, Enterococcus taecium and 2'fucosyllactose, Enterococcus taecium and GOS/FOS, Enterococcus taecium and resistant starch.

8. Composition according to claim 7, in which the combination being preferentially chosen from the group consisting of Clostridium butyricum and inulin, Clostridium butyricum and GOS/FOS, Bifidobacterium animalis lactis and inulin, Bifidobacterium animalis lactis and 2′fucosyllactose, Enterococcus taecium and beta -glucans, and Bifidobacterium crudilactis and beta-glucans, so as to allow a daily dosage of probiotic of between 1.00E+05 and 1.00E+015 CFU/day/animal, preferably between 1.00E+06 and 1.00E +013 CFU/day/animal, advantageously between 1.00E+07 and 1.00E+01 1 CFU/day/animal, and a daily prebiotic dosage of between 0.1 and 1000 g/day/animal, preferably between 0.5 and 100 g/day/animal, advantageously between 1 and 25 g/day/animal.

9. Composition according to any one of the preceding claims, containing at least one conventional excipient, in liquid or solid form.

10. Composition according to any one of the preceding claims, characterized in that said probiotic and / or said one or more prebiotics and / or said at least one conventional excipient are in encapsulated form (s) and / or in powder form and/or in the form of granule(s) and/or in liquid form, in which the said probiotic and the said one or more prebiotics of the composition are administered simultaneously, separately or staggered over time.

1 1. Composition according to any one of the preceding claims, characterized in that the therapeutically or preventively effective amount of probiotic for the treatment or prevention of dysbiosis is an amount of probiotic for

(i) reduce diarrhea in suckling and/or post-weaning piglets, and/or

(ii) increase the growth of the suckling and/or post-weaning piglet.

12. Complete or complementary feed for farm animals comprising said composition according to any one of claims 1 to 11, said feed being in the form of flour and/or granules and/or milk feeds and/or any other forms of food packaging.

13. Food for newborn animals comprising said composition according to any one of claims 1 to 1 1 and a milk base compatible with the diet of the newborn.

14. Use of a composition according to one of claims 1 to 11, to be administered to a pregnant or lactating sow, and/or to a nursing and/or post-weaning piglet for the treatment or prevention dysbiosis in suckling and/or post-weaning piglets in which the therapeutically or preventively effective quantity of probiotic is between 1.00E+05 and 100E+015 CFU/day/animal, preferably between 1.00E+06 and 1.00E+013 CFU/day/animal, advantageously between 1.00E+07 and 1.00E+01 1 CFU/day/animal, the amount of prebiotic is between 0.1 and 1000 g/day/animal , preferably between 0.5 and 100 g/day/animal, advantageously between 1 and 25 g/day/animal, said probiotic and said prebiotic of the composition being administered simultaneously, separately or staggered over time.

15. Use of a composition according to one of claims 1 to 11, for the manufacture of a medicament for the treatment or prevention of dysbiosis in suckling piglets and/or post-weaning to be administered to a sow , pregnant or lactating, and/or to a suckling and/or post-weaning piglet.

16. Method for treating or preventing dysbiosis in suckling and/or post-weaning piglets, comprising administration of a composition according to one of claims 1 to 11 to a pregnant or non-pregnant sow, for example a lactating sow, and/or lactating and/or post-weaning piglets.