WO2024126578A1

WO2024126578A1 - Synthetic composition comprising a human milk oligosaccharide for microbiota modulation

Info

Publication number: WO2024126578A1
Application number: PCT/EP2023/085585
Authority: WO
Inventors: Danica BAJIC; Frank WIENS; Pieter Arthur M VAN DEN ABBEELE
Original assignee: DSM IP Assets BV
Current assignee: DSM IP Assets BV
Priority date: 2022-12-13
Filing date: 2023-12-13
Publication date: 2024-06-20
Anticipated expiration: 2025-06-13

Abstract

The present invention relates to methods, compounds and compositions for modulating the microbiota in the gastro-intestinal tracts of humans, particularly for increasing the abundance of C. aerofaciens in the gut microbiota of humans.

Description

SYNTHETIC COMPOSITION COMPRISING A HUMAN MILK OLIGOSACCHARIDE FOR MICROBIOTA MODULATION

FIELD OF THE INVENTION

This invention relates to methods, compounds and compositions for modulating the microbiota in the gastrointestinal (Gl) tract of humans, particularly for increasing the abundance of Collinsella aerofaciens in the gut microbiota of humans.

BACKGROUND OF THE INVENTION

It has been estimated that the human Gl tract harbours 10¹³to 10¹⁴ microbial cells and the number of bacteria, archaea, and fungi exceeds the total number of cells in the body by a factor of 10. The microbiota of the human intestine and stomach is a complex and very dynamic ecosystem, which serves numerous important functions for its human host, including protection against pathogens, induction of immune regulatory functions, nutrient processing, and metabolic functions. The gut microbiota consists of various populations, which are important to preserve human health, and recent research has been able to link imbalances in the Gl microbiota to both gastrointestinal and extra-gastrointestinal inflammatory diseases.

Collinsella aerofaciens, a Gram-positive, non-spore forming, non-motile, rod-shaped obligate anaerobe, is the most abundant actinobacterium in the Gl tract of healthy humans. It is known for its ability to ferment a range of plant and animal origin carbohydrates and for producing hydrogen gas, ethanol, short-chain fatty acids, lactate, and some other metabolites in the human colon.

Experiments with an in vitro model of the human colon showed that Collinsella spp. along with Bifidobacterium spp. are the major lactose utilizers in the human gastrointestinal microbiota thereby reducing the levels of lactose in the gut. In fact, Collinsella was identified in a doubleblind study on 377 subjects as a potential key player in the beneficial gut network that increases saccharolytic potential and, hence, the ability to consume dairy products (Azcarate-Peril et al. Gut Microbes. 13, 1957536 (2021).

Moreover, Collinsella spp. take part in the deconjugation of bile acids in the gut lumen and are crucially involved in the subsequent diversification of secondary bile acids. Specifically, C. aerofaciens carries a gene encoding an NADPH-dependent 7|3-hydroxysteroid dehydrogenase (7P-HSDH), which makes it an essential intestinal bacterium to produce the secondary bile acid ursodeoxycholic acid (UDCA) from the unabsorbed bile acid chenodeoxycholic acid of liver origin (Liu et al. Appl. Microbiol. Biotechnol. 90, 127 (2011)).

UDCA has been shown to exert anti-inflammatory and protective effects in human epithelial cells of the gastrointestinal tract by signalling via cytokines, antimicrobial peptides defensins (Lajczak et al. FASEB J. 31, 3848 (2017)), and by driving faster mucosal healing in the colon (Mroz et al. Am. J. Physiol. Gastrointest. Liver Physiol. 314, G378 (2018)). Moreover, UDCA's effects include improvement of gallbladder diseases like gallstones and sclerosing cholangitis as well as primary biliary cirrhosis (Guarino et al. World J. Gastroenterol. 19, 5029 (2013)).

An altered abundance of C. aerofaciens may be linked with several health disorders, most commonly inflammatory diseases, including irritable bowel syndrome.

Several studies have observed a reduction in the amount of C. aerofaciens in the faecal samples of IBS patients compared with healthy controls.

For example, in a study done by Kassinen et al. (Gastroenterology 133, 24 (2007)), this bacterium was found in 96 % of all tested control samples, derived from subjects devoid of gastrointestinal disturbances, and its prevalence is thus well in accordance with earlier reports for this organism. In contrast, the prevalence of C. aerofaciens among patients with IBS was considerably lower.

Lyra et al. (World J. Gastroenterol. 15, 5936 (2009)) state that the relative amounts of C. aerofaciens detected were lowest in samples of the diarrhoea-predominant IBS patients. Also, as a further support to previous results, a significantly lower abundance of the C. aerofaciens was associated also with the constipation-predominant IBS subtype.

A testimony to its anti-inflammatory properties is the finding that the I L10 (potent antiinflammatory cytokine) plasma level was positively correlated with the abundances of C. aerofaciens (Zeybel et al. Adv. Sci. 9, 11 (2021)).

SARS-CoV-2 dysbiosis was also accompanied by loss of crucial bacteria that have been previously implicated in the lowering of inflammation, such as C. aerofaciens, but also Bifidobacterium adolescentis, Faecalibacterium prausnitzii, and Eubacterium rectale.

Beyond general anti-inflammatory properties of C. aerofaciens, Hirayama et al. found that Collinsella was negatively correlated with the mortality rates of COVID-19 (PLoS ONE 16, e0260451 (2021)). UDCA produced by Collinsella had been previously reported to inhibit binding of SARS-CoV-2 to angiotensin-converting enzyme 2; suppress pro-inflammatory cytokines like TNF-a, I L-ip, IL-2, IL-4, and IL-6; have antioxidant and anti-apoptotic effects; and increase alveolar fluid clearance in acute respiratory distress syndrome. Hence, Collinsella through UDCA-production may prevent COVID-19 infection and ameliorate acute respiratory distress syndrome in COVID-19 by suppressing cytokine storm syndrome.

Another study observed a negative correlation between the presence and amounts of C. aerofaciens and the BMI value of test subjects. Notably, all obese (BMI > 30) subjects in this study were negative for C. aerofaciens. Obesity has been associated with a low-grade systemic inflammation in which the Gl microbiota may be involved. This could explain the negative association observed for C. aerofaciens and BMI values.

C. aerofaciens was significantly associated with a low risk of colon cancer (Moore et al. Applied Environ. Microbiol. 61, 3202 (1995)). The incidence of C. aerofaciens showed an inverse relationship with risk of colon cancer among all the populations tested in this study (Japanese Hawaiians, North American Caucasians, rural native Japanese, rural native Africans). As persistent, low-grade inflammation is one of the causes of colon cancer, and one of the main risk factors contributing to the onset of the disease, the connection to the decrease in C. aerofaciens may point to the role this bacterium has in lowering the inflammation. UDCA is an important therapeutic agent for preventing colon cancer (Im et al. J. /Vutr. 134, 483 (2004); Khare et aL bncer Res. 63_z 3517 (2003))

Another connection to an inflammatory disease is established in a study where abundance of C. aerofaciens is shown to be significantly lower in stool of patients suffering from interstitial cystitis (IC) (Braundmeier-Fleming et al. Sc/. Rep. 6, 26083 (2016)). IC/bladder pain syndrome is associated with significant morbidity, yet underlying mechanisms and diagnostic biomarkers remain unknown. However, a central role of inflammation in IC has been confirmed in both human and animal studies.

It is important to state the finding that C. aerofaciens showed a decreasing abundance with increasing age (Stadlbauer et al. Liver Int. 40, 866 (2020)). Aging and viral infections such as SARS-CoV2, together with other factors are known inducers of gut dysbiosis. The fact that reduced C. aerofaciens was observed in both cases points to depletion of this bacterium being central to dysbiosis, which predisposes the host to a range of different disease conditions. The level of C. aerofaciens in type 2 diabetes group is significantly lower than those in normal glucose tolerance group (Zou et al. Sci. Rep. 8, 8034 (2018)).

C. aerofaciens has also been associated with diet - it has been found to be decreased in low carbohydrate weight-loss diet (Walker et al. ISME J. 5, 220 (2011)). This may be helpful for people who are struggling to find the right diet for weight-loss. Therefore, HMOs by increasing C. aerofaciens (and other beneficial bacteria that thrive on complex carbohydrates), can be the right supplement to maintain healthy microbial balance.

Human milk oligosaccharides (HMOs) are a heterogeneous mixture of soluble glycans found in human milk. They are the third most abundant solid component after lactose and lipids in human milk and are present in concentrations of 5-25 g/l (Bode: Human milk oligosaccharides and their beneficial effects, in: Handbook of dietary and nutritional aspects of human breast milk (Zibadi et al., eds.), pp. 515-31, Wageningen Academic Publishers (2013)). The structure of the HMOs is similar to the O-glycans found in mucus and the N-glycans found on human cells. HMOs are resistant to enzymatic hydrolysis in the small intestine and are thus largely undigested and unabsorbed. The majority of HMOs that reach the colon serve as substrates to shape the gut ecosystem by selectively stimulating the growth of specific bacteria. HMOs are believed to substantially modulate the infant gut microbiota and play a decisive role in the differences in the microbiota of formula-fed and breast-fed infants. However, it is not known if HMOs can impact the adult microbiota community stimulating the growth of C. aerofaciens.

There is a need, therefore, for means, preferably orally or enterally administered means, more preferably dietetic means, for effectively increasing the abundance of C. aerofaciens in the gastro-intestinal tracts of humans, preferably non-infant humans.

SUMMARY OF THE INVENTION

A first aspect of this invention relates to a human milk oligosaccharide (HMO) for use in increasing the abundance of C. aerofaciens in the gastro-intestinal tract of a mammal. Preferably, the HMO is for use in increasing the abundance of C. aerofaciens in the gastrointestinal tract of a mammal and thus to treat or prevent in the mammal diseases and/or medical conditions that are associated with depletion of C. aerofaciens in the gastrointestinal (Gl) tract of the mammal, such as: obesity, metabolic disorders associated with obesity and overweight, gallstones,

- IBS,

- IBD, interstitial cystitis,

COVID-19 infection and/or Sars-CoV-2-induced dysbiosis, age-related dysbiosis, colon cancer, lactose intolerance.

A second aspect of the invention is a synthetic composition comprising a human milk oligosaccharide (HMO) for use in increasing the abundance of C. aerofaciens in of the gastrointestinal tract of a mammal, preferably wherein the HMO is to treat or prevent in the mammal diseases and/or medical conditions that are associated with depletion of C. aerofaciens in the Gl tract of the mammal, such as: obesity, metabolic disorders associated with obesity and overweight, gallstones,

- IBS,

- IBD, interstitial cystitis,

The synthetic composition can be a nutritional or pharmaceutical composition.

A third aspect of this invention is a method for increasing the abundance of C. aerofaciens in the gastro-intestinal tract of a mammal, preferably a human, the method comprising orally or enterally administering to or providing for consumption by the mammal an effective amount of a human milk oligosaccharide (HMO). Preferably, the abundance of C. aerofaciens is increased in the gastro-intestinal track; more preferably in the colon; for example, the distal colon. Preferably, the HMO is administered to or provided for consumption by the mammal, preferably human, for a period of at least about 14 days more preferably at least about 21 days. Also, preferably, the mammal, preferably human, is administered an amount of about 1 g to about 15 g per 60 kg of body weight and per day of the HMO, more preferably about 2 g to about 10 g per day. For example, the mammal, preferably human may be administered about 3 g to about 7 g per day.

In one embodiment of the third aspect, the method comprises enterally, preferably orally, administering to or providing for consumption by a mammal, preferably human, more preferably a non-infant human:

(a) in a first step for a period of at least about 7 days (also referred to as treatment or initial treatment phase): a first amount of a human milk oligosaccharide, or a synthetic composition comprising a first amount of a human milk oligosaccharide, wherein the first amount is effective to increase the abundance of C. aerofaciens in the gastrointestinal tract of the mammal, and

(b) in a second step for an additional period, preferably of at least about 7 days (also referred to as maintenance phase): a second amount of a human milk oligosaccharide, or a synthetic composition comprising a second amount of a human milk oligosaccharide, wherein the second amount is effective to maintain the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal. Preferably, the HMO(s) is administered or provided for consumption in the first step for a period of at least about 14 days, more preferably at least about 21 days, for example up to about 28 days. Also preferably, the additional period in the second step is at least about 21 days, for example at least about 28 days. The patient may be administered higher doses during the first step and lower doses during the second step. The dose administered during the first step is preferably about 3 g to about 10 g per 60 kg of body weight and per day (for example about 4 g to about 7.5 g per day) and the dose administered during the second step is preferably about 2 g to about 7.5 g per 60 kg of body weight and per day (for example about 2 g to about 5 g per day).

A fourth aspect of the invention relates to a non-therapeutic use of a human milk oligosaccharide (HMO) for increasing the abundance of C. aerofaciens in the gastro-intestinal tract of a mammal, preferably a human. Upon administration of an effective amount of a human milk oligosaccharide orally or enterally to the mammal, the abundance of C. aerofaciens is increased, preferably, in the mucosal layer of the gastro-intestinal track; more preferably in the colon; for example, the distal colon. Preferably, the HMO is administered to or provided for consumption by the mammal, preferably human, for a period of at least about 14 days more preferably at least about 21 days. Also preferably, the mammal or human is administered an amount of about 1 g to about 15 g per 60 kg of body weight and per day of the HMO, more preferably about 2 g to about 10 g per day. For example, the mammal, preferably human may be administered about 3 g to about 7 g per day. The HMO can be comprised in a synthetic composition, e.g. in a nutritional composition.

In one embodiment of this aspect, the mammal, preferably human, more preferably a noninfant human is healthy. One target group of a healthy human is those who consume or are on a low carbohydrate diet (e.g. weight-loss diet), in order to maintain microbial diversity and microbial balance in the gut. In other embodiment, the mammal, preferably human, more preferably a non-infant human, particularly elderly, is associated with age-related dysbiosis.

In one embodiment of the fourth aspect, the method comprises enterally, preferably orally, administering to or providing for consumption by a mammal, preferably human, more preferably a non-infant human:

(b) in a second step for an additional period, preferably of at least about 7 days (also referred to as maintenance phase): a second amount of a human milk oligosaccharide, or a synthetic composition comprising a second amount of a human milk oligosaccharide, wherein the second amount is effective to maintain the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal. Preferably, the HMO(s) is administered in the first step for a period of at least about 14 days, more preferably at least about 21 days, for example up to about 28 days. Also preferably, the additional period in the second step is at least about 21 days, for example at least about 28 days. The patient may be administered higher doses during the first step and lower doses during the second step. The dose administered or provided for consumption during the first step is preferably about 3 g to about 10 g per 60 kg of body weight and per day (for example about 4 g to about 7.5 g per day) and the dose administered or provided for consumption during the second step is preferably about 2 g to about 7.5 g per 60 kg of body weight and per day (for example about 2 g to about 5 g per day).

A fifth aspect of this invention is a method for the prophylaxis or treatment of metabolic disorders associated with obesity in a mammal, the method comprising orally or enterally administering to or providing for consumption by the mammal an amount of one or more human milk oligosaccharides effective to increase the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

A sixth aspect of this invention is a method for the prophylaxis or treatment of gallstones, the method comprising orally or enterally administering to or providing for consumption by the mammal an amount of one or more human milk oligosaccharides effective to increase the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

A seventh aspect of this invention is a method for the prophylaxis or treatment of obesity in a mammal, the method comprising orally or enterally administering to or providing for consumption by the mammal an amount of one or more human milk oligosaccharides effective to increase abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

An eighth aspect of this invention is a method for the prophylaxis or treatment of an inflammation-related gastro-intestinal condition like IBD in a mammal, the method comprising orally or enterally administering to or providing for consumption by the mammal an amount of one or more human milk oligosaccharides effective to increase abundance of C. aerofaciens in the gastro-intestinal tract of the mammal. A ninth aspect of this invention is a method for the prophylaxis or treatment of IBS in a mammal, the method comprising orally or enterally administering to or providing for consumption by the mammal an amount of one or more human milk oligosaccharides effective to increase the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

A tenth aspect of this invention is a method for the prophylaxis or treatment of interstitial cystitis, the method comprising orally or enterally administering to or providing for consumption by the recipient an amount of one or more human milk oligosaccharides effective to increase the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

An eleventh aspect of this invention is a method for the prophylaxis or treatment of COVID-19 infection and/or Sars-CoV-2-induced dysbiosis, the method comprising orally or enterally administering to or providing for consumption by the recipient an amount of one or more human milk oligosaccharides effective to increase the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

A twelfth aspect of this invention is a method for the prophylaxis or treatment of age-related dysbiosis, the method comprising orally or enterally administering to or providing for consumption by the recipient an amount of one or more human milk oligosaccharides effective to increase the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

A thirteenth aspect of this invention is a method for the prophylaxis or treatment of colon cancer, the method comprising orally or enterally administering to or providing for consumption by the recipient an amount of one or more human milk oligosaccharides effective to increase the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

A fourteenth aspect of this invention is a method for the prophylaxis or treatment of lactose intolerance, the method comprising orally or enterally administering to or providing for consumption by the recipient an amount of one or more human milk oligosaccharides effective to increase the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

In any of the fifth to fourteenth aspects of the invention, the HMO is preferably administered or provided for consumption by to the mammal so as disclosed in the fourth aspect.

A fifteenth aspect of this invention relates to a pack comprising at least 14 individual daily doses of an effective amount of at least one human milk oligosaccharide (HMO) for use in increasing the abundance of C. aerofaciens in the gastro-intestinal tract of a mammal. Preferably, each dose contains about 1 g to about 15 g of the human milk oligosaccharide, more preferably about 2 g to about 10 g, for example, about 3 g to about 7 g. Preferably, the pack comprises at least 21 individual daily doses, more preferably at least 28 daily doses, for example, at least 35 daily doses. The pack can include instructions for use. The HMO is preferably for use to treat or prevent in the mammal diseases and/or medical conditions that are associated with depletion of C. aerofaciens in the gastrointestinal (Gl) tract of the mammal, such as: obesity, metabolic disorders associated with obesity and overweight, gallstones,

- IBS,

- IBD, interstitial cystitis,

COVID-19 infection and/or Sars-CoV-2-induced dysbiosis, age-related dysbiosis, colon cancer, or lactose intolerance.

Also, the pack is intended to use for a healthy mammal, preferably a human, for example for who consumes or is on a low carbohydrate diet (e.g. weight-loss diet), in order to maintain microbial diversity and microbial balance in the gut by increasing the abundance of C. aerofaciens in the Gl tract.

A sixteenth aspect of the invention is a non-therapeutical use of

- one or more human milk oligosaccharides (HMOs),

- a synthetic composition comprising one or more human milk oligosaccharides (HMOs), or

- a pack comprising at least 14 individual daily doses of an effective amount of one or more human milk oligosaccharides in the dietary management of a mammal who is healthy or suffers from one or more of the following: obesity, metabolic disorders associated with obesity and overweight, gallstones,

- IBS,

- IBD, interstitial cystitis,

COVID-19 infection and/or Sars-CoV-2-induced dysbiosis, age-related dysbiosis, colon cancer, lactose intolerance, by increasing the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal. In one embodiment, the healthy mammal, preferably human, consumes or is on a low carbohydrate diet (e.g. weight-loss diet) and the dietary management helps to maintain microbial diversity and microbial balance in the gut by increasing the abundance of C. aerofaciens in the Gl tract.

In all aspects of the invention, the HMO can be a neutral HMO or an acidic HMO. The neutral HMO can be one or more fucosylated HMOs or one or more non-fucosylated HMOs. Preferably, the HMO is 2'-FL, 3-FL, DFL, LNT, LNnT, 3'-SL, 6'-SL, LNFP-I or a mixture thereof. More preferably, the HMO comprises, consists of or consists essentially of 2'-FL, LNnT, 3'-SL, 6'-SL or any combination thereof.

In all aspects of the invention, the mammal is preferably human, more preferably a non-infant human.

BRIEF DESCRIPTION OF THE FIGURE

Figure 1 shows Collinsella aerofaciens levels (cells/m I) upon treatment with 2'-FL, LNnT, 3'-SL and 6'-SL as disclosed in Example 1 (*** indicates adjusted p-value < 0.001). DETAILED DESCRIPTION OF THE INVENTION

Herein, the following terms have the following meanings:

"About" means +/- 5 %.

"Age-related dysbiosis" represents changes in microbiota composition that can lead to a disruption in gut homeostasis in terms of integrity and overall physiology. This phenomenon is commonly referred to as dysbiosis, the term signifying the shift in the microbiota that is associated with a pathological state.

“C. aerofaciens depletion" or "Collinsella aerofaciens depletion" means a lower amount, preferably a lower relative amount, of C. aerofaciens in the microbial community of the gut of the mammal compared to the amount, preferably the relative amount, of C. aerofaciens in the microbial community of the gut of a healthy mammal.

"Colon cancer" is a disease which occurs when the cells that line the colon or the rectum become abnormal (usually due to accumulation of mutations that affect the cell-cycle) and grow out of control.

"Dietary management" means exclusive or partial feeding of subjects who, because of a disease, disorder or medical condition are suffering from:

- either have a limited, impaired or disturbed capacity to take, digest, absorb, metabolise or excrete ordinary food or certain nutrients contained therein, or metabolites, or

- have other medically-determined nutrient requirements

(see: Commission Notice on the classification of Food for Special Medical Purposes of the European Commission, Official Journal of the European Union C401, 25.11.2017, p. 10-11).

"Effective amount" means an amount of a composition that provides an HMO in a sufficient amount to render a desired treatment outcome in a human. An effective amount can be administered or provided for consumption in one or more doses to achieve the desired treatment outcome.

"Enteral administration" means any conventional form for delivery of a composition to a human that causes the deposition of the composition in the gastrointestinal tract (including the stomach). Methods of enteral administration include feeding through a naso-gastric tube or jejunum tube, oral, sublingual and rectal. "Healthy" means enjoying health, that is the condition of an organism in which it performs its vital functions normally or properly; when free from physical disease or pain and not likely to develop such.

"Human milk oligosaccharide" or "HMO" means a complex carbohydrate found in human breast milk (Urashima et al.: Milk Oligosaccharides. Nova Science Publisher (2011); Chen Adv. Carbohydr. Chem. Biochem. 72, 113 (2015)). The HMOs have a core structure comprising a lactose unit at the reducing end that can be elongated by one or more p-N-acetyl-lactosaminyl and/or one or |3-more lacto-N-biosyl units, and which core structure can be substituted by an a L-fucopyranosyl and/or an a-N-acetyl-neuraminyl (sialyl) moiety. In this regard, the non-acidic (or neutral) HMOs are devoid of a sialyl residue, and the acidic HMOs have at least one sialyl residue in their structure. The non-acidic (or neutral) HMOs can be fucosylated or non- fucosylated. Examples of such neutral non-fucosylated HMOs include lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-neohexaose (LNnH), para-lacto-N-neohexaose (pLNnH), para-lacto-N-hexaose (pLNH) and lacto-N-hexaose (LNH). Examples of neutral fucosylated HMOs include 2'-fucosyl lactose (2'-FL), lacto-N-fucopentaose I (LNFP-I), lacto-N-difucohexaose I (LNDFH-I), 3-fucosyl lactose (3-FL), difucosyllactose (DFL), lacto-N-fucopentaose II (LNFP-I I), lacto-N-fucopentaose III (LNFP-III), lacto-N-difucohexaose III (LNDFH-I II), fucosyl-lacto-N- hexaose II (FLNH-II), lacto-N-fucopentaose V (LNFP-V), lacto-N-fucopentaose VI (LNFP-VI), lacto- N-difucohexaose II (LNDFH-II), fucosyl-lacto-N-hexaose I (FLNH-I), fucosyl-para-lacto-N-hexaose I (FpLNH-l), fucosyl-para-lacto-N-neohexaose II (F-pLNnH II) and fucosyl-lacto-N-neohexaose (FLNnH). Examples of acidic HMOs include 3'-sialyllactose (3'-SL), 6'-sialyllactose (6'-SL), 3- fucosyl-3'-sialyllactose (FSL), LST a, fucosyl-LST a (FLST a), LST b, fucosyl-LST b (FLST b), LST c, fucosyl-LST c (FLST c), sialyl-LNH (SLNH), sialyl-lacto-N-hexaose (SLNH), sia lyl-lacto-N- neohexaose I (SLNH-I), sialyl-lacto-N-neohexaose II (SLNH-II) and disialyl-lacto-N-tetraose (DSLNT).

"Interstitial cystitis" is a chronic condition causing bladder pressure, bladder pain and sometimes pelvic pain. The pain ranges from mild discomfort to severe pain. The condition is a part of a spectrum of diseases known as painful bladder syndrome.

"Irritable bowel syndrome (IBS)" is a group of symptoms that occur together, including repeated pain in the abdomen and changes in bowel movements, which may be diarrhoea, constipation, or both. With IBS, these symptoms may occur without any visible signs of damage or disease in the digestive tract. "Metabolic disorder" is any disorder that disrupt normal metabolism, the process of converting food to energy on a cellular level. Metabolic disorders affect the ability of the cell to perform critical biochemical reactions that involve the processing or transport of proteins (amino acids), carbohydrates (sugars and starches), or lipids (fatty acids).

"Metabolic disorder associated with obesity or overweight" is a metabolic disorder that is likely caused by obesity, or is found in high occurrence in people who suffer from obesity. Also, a metabolic disorder for which obesity is a risk factor. Examples are type 2 diabetes, insulin resistance, hyperlipaemia.

"Microbiota", "microflora" and "microbiome" mean a community of living microorganisms that typically inhabits a bodily organ or part, particularly the gastro-intestinal organs of non-infant humans. The most dominant members of the gastrointestinal microbiota include microorganisms of the phyla of Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria, Synergistetes, Verrucomicrobia, Fusobacteria, and Euryarchaeota; at genus level Bacteroides, Faecalibacterium, Bifidobacterium, Roseburia, Alistipes, Collinsella, Blautia, Coprococcus, Ruminococcus, Eubacterium and Dorea; at species level Bacteroides uniformis, Alistipes putredinis, Parabacteroides merdae, Ruminococcus bromii, Dorea longicatena, Bacteroides caccae, Bacteroides thetaiotaomicron, Eubacterium hallii, Ruminococcus torgues, Faecalibacterium prausnitzii, Ruminococcus lactaris, Collinsella aerofaciens, Dorea formicigenerans, Bacteroides vulgatus and Roseburia intestinalis. The gastrointestinal microbiota includes the mucosa-associated microbiota, which is located in or attached to the mucus layer covering the epithelium of the gastrointestinal tract, and luminal-associated microbiota, which is found in the lumen of the gastrointestinal tract.

"Modulating of microbiota" means exerting a modifying or controlling influence on microbiota, for example an influence leading to an increase in the indigenous intestinal abundance of Bifidobacterium, Barnesiella, Faecalibacterium and/or other butyrate producing bacteria. In another example, the influence may lead to a reduction of the intestinal abundance of Ruminococcus gnavus and/or Proteobacteria. "Proteobacteria" are a phylum of Gram-negative bacteria and include a wide variety of pathogenic bacteria, such as Escherichia, Salmonella, Vibrio, Helicobacter, Yersinia and many other notable genera.

"Non-infant human" or "non-infant" means a human older than 3 years of age. A non-infant human can be a child, a teenager, an adult or an elderly person. "Non-therapeutical use" means - as opposed to the purpose of therapy which is to restore the organism from a pathological to its original condition, or to prevent pathology in the first place - an improvement of performance of the organism in a normal state. Therefore, the purpose of the non-therapeutical use is to enhance performance and perception of well-being in a healthy mammal/individual.

"Obesity" is abnormal or excessive fat accumulation that may impair health. For adults, WHO defines obesity as BMI greater than or equal to 30. For children under 5 years of age obesity is weight-for-height greater than 3 standard deviations above the WHO Child Growth Standards median. Children aged between 5-19 years obesity is BMI-for-age greater than 2 standard deviations above the WHO Growth Reference median.

"Oral administration" means any conventional form for the delivery of a composition to a human through the mouth. Accordingly, oral administration is a form of enteral administration.

"Overweight" is abnormal or excessive fat accumulation that may impair health. For adults, WHO defines overweight as a BMI greater than or equal to 25. For children under 5 years of age overweight is weight-for-height greater than 2 standard deviations above WHO Child Growth Standards median. Overweight and obesity are defined as follows for children aged between 5- 19 years overweight is BMI-for-age greater than 1 standard deviation above the WHO Growth Reference median.

"Preventive treatment" or "prevention" means treatment given or action taken to diminish the risk of onset or recurrence of a disease.

"Relative abundance" of a bacterial species means the abundance of that species relative to other bacteria in the microbiota of the gastro-intestinal tract of humans.

"Relative growth" of a bacterial species means the growth of that species relative to other bacteria in the microbiota in the gastro-intestinal tract of humans.

"Sars-CoV-2-induced dysbiosis" is an alteration in gut microbiota associated with COVID-19. The underlying mechanisms remain poorly understood. Some evidence shows that intestinal infection by SARS-CoV-2 inducing intestinal inflammation alters the gut microbiota. Another links the binding of viral S protein to angiotensin-converting enzyme 2 (ACE2) to the dysregulation of this receptor, essential in intestinal homeostasis - notably for amino acid metabolism - leading to gut dysbiosis. Additionally, SARS-CoV-2 could induce gut dysbiosis by infecting intestinal bacteria. "Secondary prevention" means prevention of onset of the condition in a high-risk patient, or prevention of reoccurrence of symptoms in a patient who has already has the condition. A "high-risk" patient is an individual who is predisposed to developing the condition; for example a person with a family history of the condition.

"Synthetic composition" means a composition which is artificially prepared and preferably means a composition containing at least one compound that is produced ex vivo chemically and/or biologically, e.g. by means of chemical reaction, enzymatic reaction or recombinantly. The synthetic composition typically comprises one or more compounds, advantageously HMOs, that are capable of preferentially increasing the abundance of C. aerofaciens in the gastrointestinal tract of the human. In some embodiments, the synthetic composition may comprise one or more compounds or components other than HMOs that may have a beneficial effect on the microbiota of a human subject microbiota in vivo, e.g. non-digestible oligosaccharides or prebiotics. Also, in some embodiments, the synthetic compositions may comprise one or more nutritionally or pharmaceutically active components which do not affect adversely the efficacy of the above-mentioned compounds. Some non-limiting embodiments of a synthetic composition of the invention are also described below.

"Therapy" means treatment given or action taken to reduce or eliminate symptoms of a disease or pathological condition.

"Treat" means to address a medical condition or disease with the objective of improving or stabilising an outcome in the person being treated or addressing an underlying nutritional need. Treat, therefore, includes the dietary or nutritional management of the medical condition or disease by addressing nutritional needs of the person being treated. "Treating" and "treatment" have grammatically corresponding meanings.

It has now been surprisingly found that administration of human milk oligosaccharides (HMOs) to humans preferentially increases the abundance of Collinsella aerofaciens, in the microbiota of their gastro-intestinal tract.

Thus, it has been discovered that human milk oligosaccharides, by oral or enteral ingestion, dynamically modulate the human intestinal microbiota by preferentially promoting the growth of Collinsella aerofaciens in addition to Bifidobacterium in the human intestine. As an outcome, a more beneficial intestinal microbial community and intestinal environment can be shaped and maintained. As a therapeutical consequence, diseases or conditions associated with C. aerofaciens depletion can be treated or prevented by the increased abundance of Collinsella aerofaciens. A disease or condition associated with Collinsella aerofaciens depletion can be for example metabolic disorders associated with obesity, obesity, an inflammation related to a gastro-intestinal condition like IBD, IBS, interstitial cystitis, Sars-CoV-2-induced dysbiosis or colon cancer. In addition, a healthy mammal or a mammal associated with age-related dysbiosis can also benefit from the increased abundance of C. aerofaciens that contributes to maintain or restore homeostasis by shaping the gut microbiota.

Concerning each aspect mentioned above, the HMOs for use in increasing the abundance of Collinsella aerofaciens in the gastro-intestinal tract of mammals may be a single HMO or a mixture of any HMOs suitable for the purpose of the invention. The HMO can be a neutral HMO or an acidic HMO. The neutral HMO is, in one embodiment, one or more fucosylated HMOs; in another embodiment, the neutral HMO is one or more non-fucosylated HMOs. Particularly, the fucosylated neutral HMO is selected from the list consisting of 2'-FL, 3-FL, DFL, LNFP-I, LNFP-II, LNFP-III, LNFP-V, LNFP-VI, LNDFH-I, LNDFH-II, LNDFH-III, FLNH-I, FLNH-II, FLNnH, FpLNH-l and F- pLNnH II, preferably 2'-FL, 3-FL, DFL or LNFP-I, more preferably 2'-FL, and the non-fucosylated neutral HMO is selected from the list consisting of LNT, LNnT, LNH, LNnH, pLNH and pLNnH, preferably LNT or LNnT, more preferably LNnT. The acidic (sialylated) HMO is selected from the list consisting of 3'-SL, 6'-SL, FSL, LST a, LST b and LST c, preferably 3'-SL and 6'-SL.

In one embodiment, the mixture comprises, consists of or essentially consists of, neutral HMOs, preferably at least a first neutral HMO and at least a second neutral HMO, wherein the first neutral HMO is a fucosylated neutral HMO as disclosed above and the second neutral HMO is a non-fucosylated neutral HMO as disclosed above. The fucosylated neutral HMO(s) and the non- fucosylated neutral HMO(s) may be present in a mass ratio of about 4:1 to 1:1. Preferably, the mixture of neutral HMOs contains, consists of or essentially consists of, a fucosylated HMO selected from the list consisting of 2'-FL, 3-FL and DFL, and a non-fucosylated neutral HMO selected from the list consisting of LNT and LNnT; advantageously the mixture comprises, consists of or essentially consists of, 2'-FL and at least one of LNnT and LNT; or at least one of 2'-FL and DFL and at least one of LNnT and LNT; or 2'-FL, DFL and at least one of LNnT and LNT; most preferably 2'-FL and LNnT.

In other embodiment, the mixture comprises, consists of or essentially consists of, at least a first (acidic) HMO as disclosed above and at least a second (neutral) HMO as disclosed above. Advantageously, the mixture comprises, consists of or essentially consists of, 2'-FL and 3'-SL; or 2'-FL and 6'-SL; LNnT and 3'-SL; LNnT and 6'SL; or 2'-FL, LNnT and 3'-SL; or 2'-FL, LNnT and 6'-SL; 2'-FL, 3'-SL and 6'-SL; or LNnT, 3'-SL and 6'-SL; or 2'-FL, LNnT, 3'-SL and 6'-SL.

The synthetic composition can be a pharmaceutical composition. The pharmaceutical composition can contain a pharmaceutically acceptable carrier, e.g. phosphate buffered saline solution, mixtures of ethanol in water, water and emulsions such as an oil/water or water/oil emulsion, as well as various wetting agents or excipients. The pharmaceutical composition can also contain other materials that do not produce an adverse, allergic or otherwise unwanted reaction when administered to humans. The carriers and other materials can include solvents, dispersants, coatings, absorption promoting agents, controlled release agents, and one or more inert excipients, such as starches, polyols, granulating agents, microcrystalline cellulose, diluents, lubricants, binders, and disintegrating agents. If desired, tablet dosages of the anti- infective compositions can be coated by standard aqueous or non-aqueous techniques.

The pharmaceutical compositions can be administered orally, e.g. as a tablet, capsule, or pellet containing a predetermined amount, or as a powder or granules containing a predetermined concentration or a gel, paste, solution, suspension, emulsion, syrup, bolus, electuary, or slurry, in an aqueous or non-aqueous liquid, containing a predetermined concentration. Orally administered compositions can include binders, lubricants, inert diluents, flavouring agents, and humectants. Orally administered compositions such as tablets can optionally be coated and can be formulated to provide sustained, delayed or controlled release of the mixture therein.

The pharmaceutical compositions can also be administered by rectal suppository, aerosol tube, naso-gastric tube or direct infusion into the Gl tract or stomach.

The pharmaceutical compositions can also include therapeutic agents such as antiviral agents, antibiotics, probiotics, analgesics, and anti-inflammatory agents. The proper dosage of these compositions for a human can be determined in a conventional manner, based upon factors such immune status, body weight and age. In some cases, the dosage will be at a concentration similar to that found for the HMO in human breast milk. The required amount would generally be in the range from about 200 mg to about 20 g per day, in certain embodiments from about 300 mg to about 15 g per day, from about 400 mg to about 10 g per day, in certain embodiments from about 500 mg to about 10 g per day, in certain embodiments from about 1 g to about 10 g per day. Appropriate dose regimes can be determined by conventional methods. The synthetic composition can also be a nutritional composition. It can contain sources of protein, lipids and/or digestible carbohydrates and can be in powdered or liquid forms. The composition can be designed to be the sole source of nutrition or a nutritional supplement. The nutritional composition is preferably for non-therapeutic use.

Suitable protein sources include milk proteins, soy protein, rice protein, pea protein and oat protein, or mixtures thereof. Milk proteins can be in the form of milk protein concentrates, milk protein isolates, whey protein or casein, or mixtures of both. The protein can be whole protein or hydrolysed protein, either partially hydrolysed or extensively hydrolysed. Hydrolysed protein offers the advantage of easier digestion which can be important for humans with inflamed Gl tracts. The protein can also be provided in the form of free amino acids. The protein can comprise about 5 % to about 30 % of the energy of the nutritional composition, normally about 10 % to 20 %.

The protein source can be a source of glutamine, threonine, cysteine, serine, proline, or a combination of these amino acids. The glutamine source can be a glutamine dipeptide and/or a glutamine enriched protein. Glutamine can be included due to the use of glutamine by enterocytes as an energy source. Threonine, serine and proline are important amino acids for the production of mucin. Mucin coats the Gl tract and can improve mucosal healing. Cysteine is a major precursor of glutathione, which is key for the antioxidant defences of the body.

Suitable digestible carbohydrates include maltodextrin, hydrolysed or modified starch or corn starch, glucose polymers, corn syrup, corn syrup solids, high fructose corn syrup, rice-derived carbohydrates, pea-derived carbohydrates, potato-derived carbohydrates, tapioca, sucrose, glucose, fructose, sucrose, lactose, honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), or mixtures thereof. Generally digestible carbohydrates provide about 35 % to about 55 % of the energy of the nutritional composition. Preferably the nutritional composition is free from lactose. A particularly suitable digestible carbohydrate is a low dextrose equivalent (DE) maltodextrin.

Suitable lipids include medium chain triglycerides (MCT) and long chain triglycerides (LCT). Preferably the lipid is a mixture of MCTs and LCTs. For example, MCTs can comprise about 30 % to about 70 % by weight of the lipids, more specifically about 50 % to about 60 % by weight. MCTs offer the advantage of easier digestion which can be important for humans with inflamed Gl tracts. Generally, the lipids provide about 35 % to about 50 % of the energy of the nutritional composition. The lipids can contain essential fatty acids (omega-3 and omega-6 fatty acids). Preferably these polyunsaturated fatty acids provide less than about 30 % of total energy of the lipid source. Decreasing the levels of these polyunsaturated fatty acids is believed to decrease sensitivity to peroxidation; which can be beneficial for humans having inflammatory conditions.

Suitable sources of long chain triglycerides are rapeseed oil, sunflower seed oil, palm oil, soy oil, milk fat, corn oil, high oleic oils, and soy lecithin. Fractionated coconut oils are a suitable source of medium chain triglycerides. The lipid profile of the nutritional composition is preferably designed to have a polyunsaturated fatty acid omega-6 (n-6) to omega-3 (n-3) ratio of about 4:1 to about 10:1. For example, the n-6 to n-3 fatty acid ratio can be about 6:1 to about 9:1.

The nutritional composition preferably also includes vitamins and minerals. If the nutritional composition is intended to be a sole source of nutrition, it preferably includes a complete vitamin and mineral profile. Examples of vitamins include vitamins A, B-complex (such as Bl, B2, B6 and B12), C, D, E and K, niacin and acid vitamins such as pantothenic acid, folic acid and biotin. Examples of minerals include calcium, iron, zinc, magnesium, iodine, copper, phosphorus, manganese, potassium, chromium, molybdenum, selenium, nickel, tin, silicon, vanadium and boron.

The nutritional composition can also include a carotenoid such as lutein, lycopene, zeaxanthin, and beta-carotene. The total amount of carotenoid included can vary from about 0.001 pg/ml to about 10 pg/ml. Lutein can be included in an amount of from about 0.001 pg/ml to about 10 pg/ml, preferably from about 0.044 pg/ml to about 5 g/ml of lutein. Lycopene can be included in an amount from about 0.001 pg/ml to about 10 pg/ml, preferably about 0.0185 mg/ml to about 5 g/ml of lycopene. Beta-carotene can comprise from about 0.001 pg/ml to about 10 mg/ml, for example about 0.034 pg/ml to about 5 pg/ml of beta-carotene.

The nutritional composition preferably also contains reduced concentrations of sodium; for example, from about 300 mg/l to about 400 mg/l. The remaining electrolytes can be present in concentrations set to meet needs without providing an undue renal solute burden on kidney function. For example, potassium is preferably present in a range of about 1180 to about 1300 mg/l; and chloride is preferably present in a range of about 680 to about 800 mg/l.

The nutritional composition can also contain various other conventional ingredients such as preservatives, emulsifying agents, thickening agents, buffers, fibres and prebiotics (e.g. fructooligosaccharides, galactooligosaccharides), probiotics (e.g. B. animalis subsp. lactis BB-12, B. lactis HN019, B. lactis Bi07, B. infantis ATCC 15697, L. rhamnosus GG, L. rhamnosus HNOOI, L. acidophilus LA-5, L. acidophilus NCFM, L. fermentum CECT5716, B. longum BB536, B. longum AH1205, B. longum AH1206, B. breve M-16V, L. reuteri ATCC 55730, L. reuteri ATCC PTA-6485, L. reuteri DSM 17938), antioxidant/anti-inflammatory compounds including tocopherols, carotenoids, ascorbate/vitamin C, ascorbyl palmitate, polyphenols, glutathione, and superoxide dismutase (melon), other bioactive factors (e.g. growth hormones, cytokines, TFG- ), colorants, flavours, and stabilisers, lubricants, and so forth.

The nutritional composition can be in the form of a soluble powder, a liquid concentrate, or a ready-to-use formulation. The composition can be fed to a human via a nasogastric tube or orally. Various flavours, fibres and other additives can also be present.

The nutritional compositions can be prepared by any commonly used manufacturing techniques for preparing nutritional compositions in solid or liquid form. For example, the composition can be prepared by combining various feed solutions. A protein-in-fat feed solution can be prepared by heating and mixing the lipid source and then adding an emulsifier (e.g. lecithin), fat soluble vitamins, and at least a portion of the protein source while heating and stirring. A carbohydrate feed solution is then prepared by adding minerals, trace and ultratrace minerals, thickening or suspending agents to water while heating and stirring. The resulting solution is held for 10 minutes with continued heat and agitation before adding carbohydrates (e.g. the HMOs and digestible carbohydrate sources). The resulting feed solutions are then blended together while heating and agitating and the pH adjusted to 6.6-7.0, after which the composition is subjected to high-temperature short-time processing during which the composition is heat treated, emulsified and homogenized, and then allowed to cool. Water soluble vitamins and ascorbic acid are added, the pH is adjusted to the desired range if necessary, flavours are added, and water is added to achieve the desired total solid level.

For a liquid product, the resulting solution can then be aseptically packed to form an aseptically packaged nutritional composition. In this form, the nutritional composition can be in ready-to- feed or concentrated liquid form. Alternatively, the composition can be spray-dried and processed and packaged as a reconstitutable powder.

When the nutritional product is a ready-to-feed nutritional liquid, the total concentration of HMOs in the liquid, by weight of the liquid, is from about 0.0001 % to about 2.0 %, including from about 0.001 % to about 1.5 %, including from about 0.01 % to about 1.0 %. When the nutritional product is a concentrated nutritional liquid, the total concentration of HMOs in the liquid, by weight of the liquid, is from about 0.0002 % to about 4.0 %, including from about 0.002 % to about 3.0 %, including from about 0.02 % to about 2.0 %.

The nutritional composition can also be in a unit dosage form such as a capsule, tablet or sachet. For example, the synthetic composition can be in a tablet form comprising the HMOs, and one or more additional components to aid formulation and administration, such as diluents, excipients, antioxidants, lubricants, colorants, binders, disintegrants, and the like.

Suitable diluents, excipients, lubricants, colorants, binders, and disintegrants include polyethylene, polyvinyl chloride, ethyl cellulose, acrylate polymers and their copolymers, hydroxyethyl-cellulose, hydroxypropylmethyl-cellulose (HPMC), sodium carboxymethylcellulose, polyhydroxyethyl methylacrylate (PHEMA), polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), polyethylene oxide (PEO), or polyacrylamide (PA), carrageenan, sodium alginate, polycarbophil, polyacrylic acid, tragacanth, methyl cellulose, pectin, natural gums, xanthan gum, guar gum, karaya gum, hypromellose, magnesium stearate, microcrystalline cellulose, and colloidal silicon dioxide. Suitable antioxidants are vitamin A, carotenoids, vitamin C, vitamin E, selenium, flavonoids, polyphenols, lycopene, lutein, lignan, coenzyme Q10 ("CoQIO") and glutathione.

The unit dosage forms, especially those in sachet form, can also include various nutrients including macronutrients.

A first target group of this invention includes mammals, preferably humans having depleted amount of Collinsella aerofaciens in their gut but they are otherwise healthy. Their ingestion of one or more HMOs will stimulate the growth of Collinsella aerofaciens in the gastro-intestinal tract and increase the abundance of Collinsella aerofaciens in the gastro-intestinal tract.

A second target group for this invention includes mammals, preferably humans, more preferably elderly who are associated with age-related dysbiosis. Their ingestion of one or more HMOs will stimulate the growth of Collinsella aerofaciens in the gastro-intestinal tract and increase abundance of Collinsella aerofaciens in the gastro-intestinal tract.

A third target group for this invention includes mammals, preferably humans having depleted amount of Collinsella aerofaciens in their gut and suffering from one or more of: metabolic disorders associated with obesity and overweight, gallstones, obesity, IBS, IBD, interstitial cystitis, COVID-19 infection and/or Sars-CoV-2-induced dysbiosis, age-related dysbiosis, colon cancer, lactose intolerance. Their ingestion of one or more HMOs will stimulate the growth of Collinsella aerofaciens in the gastro-intestinal tract and increase abundance of Collinsella aerofaciens in the gastro-intestinal tract.

The HMO can be administered to or provided for consumption by the human:

(a) in a first step, for a period of up to about 14 days: a first amount of a human milk oligosaccharide, or a synthetic composition comprising a first amount of a human milk oligosaccharide, to increase the abundance of Collinsella aerofaciens in the gastro-intestinal tract of the human to the level up to 100 % or more, such as 200-500 % higher, compared to the abundance of Collinsella aerofaciens before the initiation of administration, and

(b) in a second step, for an additional period: a second amount of a human milk oligosaccharide, or a synthetic composition comprising a second amount of a human milk oligosaccharide, to maintain the level of Collinsella aerofaciens in the gastro-intestinal tract of the human achieved after the first step.

For stimulating the growth of Collinsella aerofaciens in the gastro-intestinal tract of a human, the amount of HMO(s) required to be administered or provided for consumption will vary depending upon factors such as the risk and severity of the diseases associated with depletion of C. aerofaciens mentioned above, age, the form of the composition, and other medications being administered. However, the required amount can be readily set by a medical practitioner and would generally be in the range from about 10 mg to about 20 g per day, in certain embodiments from about 10 mg to about 15 g per day, from about 100 mg to about 10 g per day, in certain embodiments from about 500 mg to about 10 g per day, in certain embodiments from about 1 g to about 7.5 g per day. An appropriate dose can be determined based on several factors, including, for example, body weight and/or condition, the severity of type 2 diabetes, the inflammatory gastrointestinal condition or the enteropathogenic infection, being treated or prevented, other ailments and/or diseases, the incidence and/or severity of side effects and the manner of administration. Appropriate dose ranges may be determined by methods known to those skilled in the art. During an initial treatment phase (first step), the dosing can be higher (for example 200 mg to 20 g per day, preferably 500 mg to 15 g per day, more preferably 1 g to 10 g per day, in certain embodiments 2.5 g to 7.5 g per day). During a maintenance phase (second step), the dosing can be reduced (for example, 10 mg to 10 g per day, preferably 100 mg to 7.5 g per day, more preferably 500 mg to 5 g per day, in certain embodiments 1 g to 2.5 g per day).

Whilst the invention has been described with reference to a preferred embodiment, it will be appreciated that various modifications are possible within the scope of the invention.

EXAMPLES

The working example described herein are for illustration purposes only and should not be considered as limiting.

Example 1

A kinetic, ex vivo study was implemented, simulating the colonic fermentation of test products by the gut microbiota derived from healthy human adults (around 30 years old) (n = 6). Individual bioreactors were processed in parallel in a bioreactor management device (Cryptobiotix, Ghent, Belgium). Each bioreactor contained 5 ml of nutritional medium-faecal inoculum blend dosed with 5 g HMO/I, sealed individually, before being rendered anaerobic. After preparation, bioreactors were incubated under continuous agitation (140 rpm) at 37 °C for 24 h. Upon gas pressure measurement in the headspace, liquid samples were collected for subsequent analysis. Test products were HMOs (2'-FL, LNnT, 3'-SL, 6'-SL), at the concentration 5 g/l, simulating intake of 5 g per day.

In this study, we have shown that all HMOs tested increased abundance of C. aerofaciens. The abundance was determined by shallow shotgun sequencing. Upon DNA extraction, standardized Illumina library preparation was performed followed by 3M total DNA sequencing. Quantitative data was obtained by correcting relative abundances (%) with total cell counts for each sample (cells/m I; flow cytometry), resulting in cell counts/ml. This cell count/ml value represents absolute abundance of C. aerofaciens. Sequencing protocol: DNA was extracted via the SPINeasy DNA Kit for Soil (MP Biomedicals, Eschwege, Germany), according to manufacturer's instructions. Subsequently, DNA libraries were prepared using the Nextera XT DNA Library Preparation Kit (Illumina, San Diego, CA, United States) and IDT Unique Dual Indexes with total DNA input of Ing. Genomic DNA was fragmented using a proportional amount of Illumina Nextera XT fragmentation enzyme. Unique dual indexes were added to each sample followed by 12 cycles of PCR to construct libraries. DNA libraries were purified using AMpure magnetic Beads (Beckman Coulter, Brea, CA, United States), eluted in QIAGEN EB buffer, quantified using a Qubit 4 fluorometer and a Qubit dsDNA HS Assay Kit, and sequenced on an Illumina Nextseq 2000 platform 2xl50bp. Unassembled sequencing reads were converted to relative abundances (%) using the CosmosID-HUB Microbiome Platform (CosmosID Inc., Germantown, MD, United States). For total cell count analysis, liquid samples were diluted in anaerobic phosphate-buffered saline (PBS), after which cells were stained with SYTO 16 at a final concentration of lpM and counted via a BD FACS Verse flow cytometer (BD, Erembodegem, Belgium). Data was analysed using FlowJo, version 10.8.1.

C. aerofaciens levels were markedly increased upon treatment with 2'-FL, LNnT, 3'-SL and 6'-SL (Figure 1). These increases were statistically significant (based on repeated measures one-way ANOVA with posthoc Benjamini Hochberg correction (FDR = 0.05) on loglO-transformed abundances) with adjusted p-values being below 0.001 for 2'-FL, LNnT, 3'-SL and 6'-SL.

Claims

1. A non-therapeutic use of a human milk oligosaccharide (HMO) for increasing the abundance of C. aerofaciens in the gastro-intestinal tract of a mammal.

2. A non-therapeutical use of one or more human milk oligosaccharides (HMOs), a synthetic composition comprising one or more human milk oligosaccharides (HMOs), or a pack comprising at least 14 individual daily doses of an effective amount of one or more human milk oligosaccharides in the dietary management of a mammal who is healthy or suffers from one or more of the following: obesity, metabolic disorders associated with obesity and overweight, gallstones,

IBS,

IBD, interstitial cystitis,

COVID-19 infection and/or Sars-CoV-2-induced dysbiosis, age-related dysbiosis, colon cancer, lactose intolerance, by increasing the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

3. The use according to claim 1 or 2, wherein the mammal is a healthy human.

4. The use according to claim 3, wherein the human consumes or is on a low carbohydrate diet.

5. The use according to any of the claims 1 to 4, wherein the HMO is a fucosylated neutral HMO, preferably 2'-FL, 3-FL and/or DFL, more preferably 2'-FL.

6. The use according to any of the claims 1 to 4, wherein the HMO is a non-fucosylated neutral HMO, preferably LNT and/or LNnT, more preferably LNnT.

7. The use according to any of the claims 1 to 4, wherein the HMO is a sialylated HMO, preferably 3'-SL and/or 6'-SL.

8. A human milk oligosaccharide (HMO) or a synthetic composition comprising an HMO for use in increasing the abundance of Collinsella aerofaciens in the gastro-intestinal tract of a mammal.

9. The HMO or the synthetic composition for use according to claim 7 to treat or prevent in the mammal diseases and/or medical conditions that are associated with depletion of C. aerofaciens in the gastrointestinal (Gl) tract of the mammal.

10. The HMO or the synthetic composition for use according to claim 8 or 9 to treat or prevent in the mammal obesity, metabolic disorders associated with obesity and overweight, gallstones,

IBS,

IBD, interstitial cystitis,

11. The HMO or the synthetic composition for use according to any of the claims 8 to 10, wherein the mammal is a human, preferably a non-infant human.

12. The HMO or the synthetic composition for use according to any of the claims 8 to 11, wherein the HMO is a fucosylated neutral HMO, preferably 2'-FL, 3-FL and/or DFL, more preferably 2'- FL, a non-fucosylated neutral HMO, preferably LNT and/or LNnT, more preferably LNnT, and/or a sialylated HMO, preferably 3'-SL and/or 6'-SL.

13. A method for increasing the abundance of C. aerofaciens in the gastro-intestinal tract of a mammal, preferably a human, the method comprising orally or enterally administering to or providing for consumption by the mammal an effective amount of a human milk oligosaccharide (HMO).

14. A method for the prophylaxis or treatment of obesity, metabolic disorders associated with obesity and overweight, gallstones,

IBS, an inflammation-related gastro-intestinal condition like IBD, interstitial cystitis,

COVID-19 infection and/or Sars-CoV-2-induced dysbiosis, age-related dysbiosis, colon cancer, or lactose intolerance in a mammal, the method comprising orally or enterally administering to or providing for consumption by the mammal an amount of one or more human milk oligosaccharides effective to increase the abundance of C. aerofaciens in the gastro-intestinal tract of the mammal.

15. The method according to claim 13 or 14, wherein the mammal is a human, preferably a non-infant human.

16. The method according to any of the claims 13 to 15, wherein the HMO is a fucosylated neutral HMO, preferably 2'-FL, 3-FL and/or DFL, more preferably 2'- FL, a non-fucosylated neutral HMO, preferably LNT and/or LNnT, more preferably LNnT, and/or a sia lylated HMO, preferably 3'-SL and/or 6'-SL.