Field of the invention

The present invention relates to a fucosylated saccharide for use in a method for preventing and/or treating a bacterial infection in a subject. More specifically, the present invention relates to a fucosylated saccharide for use in a method for preventing and/or treating streptococcosis.

Background of the invention

Bacteria are responsible for a replete amount of diseases among humans and animals, in particular in young or immunocompromised subjects. One of the most common bacterial disease found worldwide is streptococcosis. It is a bacterial disease which is caused by Streptococcus. Members of the genus Streptococcus cause mild to severe bacterial illnesses in humans and animals. Streptococcus is a grampositive bacterium and typically colonizes one or more species as commensals and can cause opportunistic infections in those hosts. Streptococcus strains are not completely host-specific. Streptococcus suis for example is normally found in pigs, but has emerged as a significant agent of human illnesses. Another example is Streptococcus agalactiae which is a fish-associated strain that can affect humans as well ("Zoonotic Streptococcosis", updated September 2020 and accessible on https://www.cfsph.iastate.edu/Factsheets/pdfs/ streptococcosis.pdf). Many streptococcal species are multi-host pathogens. Streptococcus agalactiae, Streptococcus iniae and Streptococcus dysgalactiae do not only infect fish, but are also well-known human pathogens and hence impose a major threat to public health. Streptococcus agalactiae for example can cause meningitis and pneumonia in humans, while Streptococcus iniae can cause meningitis, endocarditis and septic arthritis.

Aquaculture is one of the fastest growing businesses in the food industry, but is confronted with significant economic losses due to streptococcal infections. Fish are an important food source, but have always been at risk of acquiring Streptococcus infections due to the continuous exposure and ubiquitous global presence of various Streptococcus strains. The most prominent ones in fish include Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae, Sreptococcus phocae and Streptococcus parauberis (Mishra et al, 2017; Morales-Covarrubias et al, 2018), while shrimp are found to be infected with Streptococcus penaeicida (Morales-Covarrubias et al, 2018).

Antimicrobial compounds, vaccinations and environmental strategies are currently applied to control Streptococcus infections, but appear to be ineffective for various reasons (Mishra et al, 2017). Liquid disinfecting agents are a valuable option as Streptococcus initially affects skin, fins, gills and external organs. However, these agents are hazardous for the environment. Antibiotics such as erythromycin, penicillin and tetracycline can be efficient, however, the main disadvantage of their use lies in the well- known phenomenon of antibiotic resistance which forms a major concern associated with human health.

Vaccinations strategies are currently unsuccessful due to the poorly studied immunity in fish.

The development of an effective therapeutic for preventing and/or treating bacterial disease such as streptococcosis, is a medical and public health imperative.

Summary of the invention

It was surprisingly found that a fucosylated saccharide according to the invention proves to be an efficacious agent to prevent and/or treat a bacterial infection, in particular streptococcosis. Said fucosylated saccharide is not associated with adverse effects and is safe to use in humans and animals.

In a first aspect, the invention provides a saccharide comprising a fucose (i.e. fucosylated saccharide) for use in a method for preventing and/or treating a bacterial infection in a subject.

In a second aspect, the invention provides a composition comprising a fucosylated saccharide for use in a method for preventing and/or treating a bacterial infection in a subject.

In a third aspect, the invention provides a method for preventing and/or treating a bacterial infection in a subject, wherein a fucosylated saccharide or a composition comprising a fucosylated saccharide is administered to a subject.

In a fourth aspect, the invention provides the use of a fucosylated saccharide or composition comprising a fucosylated saccharide for the manufacture of a medicament for preventing and/or treating a bacterial infection in a subject.

Detailed description of the invention

Saccharide for use in preventing/treating a bacterial infection

In a first aspect, the invention provides a saccharide comprising a fucose for use in a method for preventing and/or treating a bacterial infection in a subject.

In the context of the present invention, the term "preventing" a bacterial infection or a disease such as streptococcosis as described herein, preferably means avoiding that said bacterial infection or disease occurs and/or decreasing the incidence of said infection or disease. In other words, "preventing" preferably refers to ameliorating the risk of suffering from said bacterial infection or said disease. The term "prevention" and "prophylaxis" are interchangeably used in the context of the present invention. The term "treating" a bacterial infection or a disease such as streptococcosis as described herein, preferably means inhibiting said infection or disease, e.g. arresting the development of said infection or disease; relieving said infection or disease, for example causing regression of said infection or disease; and/or relieving a condition caused by or resulting from said infection or disease, e.g. relieving, preventing or treating symptoms of said infection or disease. In other words, "treating" preferably refers to decreasing the duration (number of days/weeks/months/years) the subject will suffer from said infection or disease), the risks, the complications and/or the severity of said infection or disease; this also encompasses the relief of the symptoms caused by said infection or disease.

Saccharide

In an embodiment of the first aspect of the invention, said saccharide comprises a fucose. Preferably, said saccharide comprises only one fucose, i.e. said saccharide comprises one or more monosaccharides and only one of said monosaccharides is a fucose. As understood by the skilled person, the term "saccharide" refers to a molecule comprising at least one monosaccharide, i.e. a saccharide is a molecule consisting of one or more monosaccharide residue(s). The term "monosaccharide" as used herein refers to a sugar that is not decomposable into simpler sugars by hydrolysis, is classed either an aldose or ketose, and contains one or more hydroxyl groups per molecule. Monosaccharides are saccharides containing only one simple sugar.

In an additional and/or alternative embodiment, said saccharide is a neutral saccharide. A "neutral" saccharide as used herein and as generally understood in the state of the art is a saccharide that has no negative charge originating from a carboxylic acid group.

In an additional and/or alternative embodiment, said fucose of a saccharide according to the invention is linked to a monosaccharide in an alpha-1,2-, alpha-1,3- or alpha-1, 4-linkage, preferably an alpha-1,2- or an alpha-1, 3-linkage, more preferably an alpha-1, 3-linkage, and wherein said monosaccharide is preferably selected from glucose, N-acetylglucosamine and galactose, more preferably said monosaccharide is glucose or N-acetylglucosamine, even more preferably said monosaccharide is glucose. The skilled person will understand that the expression "fucose is linked to a monosaccharide" refers to the situation wherein the fucose is bound to a monosaccharide through a glycosidic bond and wherein said fucose and monosaccharide are part of the saccharide of the invention (which can comprise additional monosaccharide(s) than said fucose and said monosaccharide unless said saccharide is a disaccharide).

In a preferred embodiment, said saccharide is a disaccharide or an oligosaccharide. In a more preferred embodiment, said saccharide is an oligosaccharide. In the context of the present invention, the term "oligosaccharide" preferably refers to a saccharide containing 2 up to and including 20 monosaccharides, i.e. the degree of polymerization (DP) is 2-20. An oligosaccharide can be a linear structure or can include branches. The linkage (e.g. glycosidic linkage, galactosidic linkage, glucosidic linkage, etc.) between two sugar units can be expressed, for example, as 1,4, l->4, or (1-4), used interchangeably herein. Each monosaccharide can be in the cyclic form (e.g. pyranose or furanose form). An oligosaccharide can contain both alpha- and beta-glycosidic bonds or can contain only beta-glycosidic bonds.

More preferably, said oligosaccharide consists of 3-9, preferably 3-8, more preferably 3-7, even more preferably 3-6, even more preferably 3-5, most preferably 3 or 4, monosaccharides.

In an additional and/or alternative preferred embodiment, said saccharide according to the invention is a mammalian milk oligosaccharide (MMO), preferably a human milk oligosaccharide (HMO). Mammalian milk oligosaccharides (MMOs) comprise oligosaccharides present in milk found in any phase during lactation including colostrum milk from humans (i.e. human milk oligosaccharides or HMOs) and mammals including but not limited to cows (Bos Taurus), sheep (Ovis aries), goats (Capra aegagrus hircus), bactrian camels (Camelus bactrianus), horses (Eguus ferus caballus), pigs (Sus scropha), dogs (Canis lupus familiaris), ezo brown bears (Ursus arctos yesoensis), polar bear (Ursus maritimus), Japanese black bears (Ursus thibetanus japonicus), striped skunks (Mephitis mephitis), hooded seals (Cystophora cristata), Asian elephants (Elephas maximus), African elephant (Loxodonta africana), giant anteater (Myrmecophaga tridactyla), common bottlenose dolphins (Tursiops truncates), northern minke whales (Balaenoptera acutorostrata), tammar wallabies (Macropus eugenii), red kangaroos (Macropus rufus), common brushtail possum (Trichosurus Vulpecula), koalas (Phascolarctos cinereus), eastern quolls (Dasyurus viverrinus), platypus (Ornithorhynchus anatinus). Human milk oligosaccharides (HMOs) are also known as human identical milk oligosaccharides which are chemically identical to the human milk oligosaccharides found in human breast milk but which are biotechnologically-produced (e.g. using cell free systems or cells and organisms comprising a bacterium, a fungus, a yeast, a plant, animal, or protozoan cell, preferably genetically engineered cells and organisms). Human identical milk oligosaccharides are marketed under the name HiMO.

In an additional and/or alternative preferred embodiment, said saccharide according to the invention comprises a lactose, a lacto-N-biose (LNB) or N-acetyllactosamine (LacNAc) at its reducing end, preferably said saccharide according to the invention comprises lactose or LacNAc at is reducing end, more preferably said saccharide according to the invention comprises lactose at its reducing end.

In an additional and/or alternative preferred embodiment, said saccharide according to the invention is selected from a list consisting of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3-FL), difucosyllactose (diFL), 2'-fucosyl-N-acetyllactosamine (2'FlacNAc), difucosyl-N-acetyllactosamine (diFLacNAc), 3-fucosyl-N- acetyllactosamine (3FlacNAc), 2'-fucosyllacto-N-biose (2'FLNB), 4-fucosyllacto-N-biose (4FLNB), difucosyllacto-N-biose (diFLNB), lacto-N-fucopentaose I (LNFP I), blood group A antigen hexaose type 1 (GalNAc-LNFP I), blood group B antigen hexaose type 1 (Gal-LNFP I), lacto-N-fucopentaose II (LNFP ll) ₇ lacto-N-fucopentaose III (LNFP III), lacto-N-fucopentaose V (LNFP V), lacto-N-difucohexaose I (LNDFH I), lacto-N-difucohexaose II (LNDFH II), lewis b-lewis x, monofucosyllacto-N-hexaose III (MFLNH III), difucosyllacto-N-hexaose (a) (DFLNH (a)), difucosyllacto-N-hexaose (DFLNH), trifucosyllacto-N-hexaose (TFLNH), lacto-N-neofucopentaose I (LNnFP I), lacto-N-neofucopentaose V (LNnFP V, LNFP VI), and lacto- N-neodifucohexaose (LNnDFH), preferably selected from a list consisting of 2'FL, 3-FL, diFL, 2'FlacNAc, diFLacNAc, 3FlacNAc, 2'FLNB, 4FLNB, diFLNB, LNFP II, LNFP III, LNFP V, LNDFH I, lewis b-lewis x, MFLNH III, DFLNH (a), DFLNH, TFLNH and LNnFP I. Said saccharides are commercially available and/or the production/purification of any of said saccharides has been described and allows the skilled person to produce/obtain any of said saccharides accordingly. An exemplary reference is provided for each saccharide and each reference is incorporated by reference:

- 2'FL: Zhou et al, 2021; Lu et al, 2021 ; W02022/034073; Carbosynth (OF06739)

- 3-FL: Zhou et al, 2021; Lu et al, 2021 ; W02022/034073; Carbosynth (OF05673)

- diFL: W02022/034073

- 2'FlacNAc: WO 2022/034075

- 3FlacNAc: WO 2022/034075

- diFLacNAc: WO 2022/034075

- 2'FLNB: WO 2022/034075

- 4FLNB: WO 2022/034075

- diFLNB: WO 2022/034075

- LNFP I: Derya et al, 2020 ; Carbosynth (OL05676)

- GalNAc-LNFP I: WO 2022/034077

- Gal-LNFP I: WO 2022/034077

- LNFP II: Zeuner et al, 2018

- LNFP III: Zeuner et al, 2018 ; Carbosynth (OL04212)

- LNFP V: W02020/115671; Carbosynth (OL06817)

- LNDFH I: Huang et al, 2021; Carbosynth (OL01664)

- LNDFH II: Yu et al, 2017 ; Huang et al, 2021 ; Carbosynth (OL06826)

- MFLNH III: Carbosynth (OM05898)

- DFLNH (a): Carbosynth (OD05375)

- DFLNH: Carbosynth (OD06532)

- TFLNH: Isosep (57/18-0010)

- LNnFP I: Elicityl (GLY033-2-90%)

LNnFP V: Dumon et al, 2001

LNnDFH: Dumon et al, 2001 In an additional and/or alternative preferred embodiment, said saccharide according to the invention is selected from a list consisting of 3-FL, diFL, LNFP III, LNFP V, LNDFH II, lewis b-lewis x, MFLNH III, DFLNH (a), DFLNH, TFLNH, LNnFP V, LNnDFH, 3FlacNAc and diFLacNAc, preferably selected from a list consisting of 3-FL, diFL, LNFP V, LNDFH II, lewis b-lewis x, LNnFP V, LNnDFH, 3FlacNAc and diFLacNAc, more preferably selected from a list consisting of 3-FL, diFL, LNDFH II, lewis b-lewis x, LNnFP V, LNnDFH, 3FlacNAc and diFLacNAc, even more preferably selected from a list consisting of 3-FL, diFL, 3FlacNAc and diFLacNAc, even more preferably wherein said saccharide is 3-FL or 3FlacNAc, even more preferably wherein said saccharide is 3-FL or diFL, most preferably wherein said saccharide is 3-FL.

In an additional and/or alternative preferred embodiment, said saccharide according to the invention is selected from a list consisting of 3-FL, 3FlacNAc, LNFP-III, MFLNH III, DFLNH (a), DFLNH and TFLNH, preferably selected from a list consisting of LNFP-III, MFLNH III, DFLNH (a), DFLNH and TFLNH, more preferably wherein said oligosaccharide is LNFP-III.

In an additional and/or alternative preferred embodiment, said saccharide according to the invention is not LNFP V. In an additional and/or alternative preferred embodiment, said saccharide according to the invention is not LNFP I. In an additional and/or alternative preferred embodiment, said saccharide according to the invention is not LNDFH II. In an additional and/or alternative preferred embodiment, said saccharide according to the invention is not LNnDFH. In an additional and/or alternative preferred embodiment, said saccharide according to the invention is not LNnFP V.

In another embodiment, said saccharide of the invention has been isolated from a microbial cultivation or fermentation, cell culture, enzymatic reaction or chemical reaction. Alternatively, said saccharide of the invention has been isolated by e.g. chromatography or filtration technology from a natural source such as a human or animal milk, preferably animal milk. For example Lu et al, 2021, provides a review on methods used for the production of 3-fucsyllactose, an exemplary saccharide according to the invention, and which is incorporated in its entirety herein.

In a preferred embodiment, said saccharide of the invention has been produced, preferably in vitro and/or ex vivo, by a cell, preferably a single cell, wherein said cell is preferably chosen from the list consisting of a microorganism, a plant cell, an animal cell or a protozoan cell. In other words, said saccharide of the invention has been produced by an in vitro and/or ex vivo culture of cells, wherein said cells are preferably chosen from the list consisting of a microorganism, a plant cell, an animal cell or a protozoan cell.

Preferably, said cell is a microorganism. Preferably, said microorganism is selected from a list consisting of a bacterium, a yeast or a fungus. Further, it is preferred that said cell is genetically engineered for the production of said saccharide according to the invention. Throughout the description and claims, the terms "has been isolated" and "is obtained" are interchangeably used, unless specifically stated otherwise.

The latter bacterium preferably belongs to the phylum of the Proteobacteria or the phylum of the Firmicutes or the phylum of the Cyanobacteria or the phylum Deinococcus-Thermus, preferably belongs to the phylum of the Proteobacteria. The latter bacterium belonging to the phylum Proteobacteria belongs preferably to the family Enterobacteriaceae, preferably to the species Escherichia coli. The latter bacterium preferably relates to any strain belonging to the species Escherichia coli such as but not limited to Escherichia coli B, Escherichia coli C, Escherichia coli W, Escherichia coli K12, Escherichia coli Nissle. More specifically, the latter term relates to cultivated Escherichia coli strains - designated as E. coli K12 strains - which are well-adapted to the laboratory environment, and, unlike wild type strains, have lost their ability to thrive in the intestine. Well-known examples of the E. coli K12 strains are K12 Wild type, W3110, MG1655, M182, MC1000, MC1060, MC1061, MC4100, JM101, NZN111 and AA200. Hence, preferably the present invention specifically relates to a mutated and/or transformed Escherichia coli strain as indicated above wherein said E. coli strain is a K12 strain. More specifically, the present invention relates to a mutated and/or transformed Escherichia coli strain as indicated above wherein said K12 strain is E. coli MG1655. The latter bacterium belonging to the phylum Firmicutes belongs preferably to the Bacilli, preferably from the species Bacillus, such as Bacillus subtilis or, B. amyloliquefaciens. The latter Bacterium belonging to the phylum Actinobacteria, preferably belonging to the family of the Corynebacteriaceae, with members Corynebacterium glutamicum or C. afermentans, or belonging to the family of the Streptomycetaceae with members Streptomyces griseus or S. fradiae. The latter yeast preferably belongs to the phylum of the Ascomycota or the phylum of the Basidiomycota or the phylum of the Deuteromycota or the phylum of the Zygomycetes. The latter yeast belongs preferably to the genus Saccharomyces (with members like e.g. Saccharomyces cerevisiae, S. bayanus, S. boulardii), Pichia (with members like e.g. Pichia pastoris, P. anomala, P. kluyveri), Komagataella, Hansunella, Kluyveromyces (with members like e.g. Kluyveromyces lactis, K. marxianus, K. thermotolerans), Yarrowia (like e.g. Yarrowia lipolytica), Eremothecium, Zygosaccharomyces, Starmerella fl ike e.g. Starmerella bombicola) or Debaromyces. The latter yeast is preferably selected from Pichia pastoris, Yarrowia lipolitica, Saccharomyces cerevisiae and Kluyveromyces lactis. The latter fungus belongs preferably to the genus Rhizopus, Dictyostelium, Penicillium, Mucor or Aspergillus. "Plant cells" includes cells of flowering and nonflowering plants, as well as algal cells, for example Chlamydomonas, Chlorella, etc. Preferably, said plant cell is a tobacco, alfalfa, rice, cotton, rapeseed, tomato, corn, maize or soybean cell. The latter animal cell is preferably derived from non-human mammals (e.g. cattle, buffalo, pig, sheep, mouse, rat), birds (e.g. chicken, duck, ostrich, turkey, pheasant), fish (e.g. swordfish, salmon, tuna, sea bass, trout, catfish), invertebrates (e.g. lobster, crab, shrimp, clams, oyster, mussel, sea urchin), reptiles (e.g. snake, alligator, turtle), amphibians (e.g. frogs) or insects (e.g. fly, nematode) or is a genetically modified cell line derived from human cells excluding embryonic stem cells. Both human and non-human mammalian cells are preferably chosen from the list comprising an epithelial cell like e.g. a mammary epithelial cell, mammary myoepithelial cell, mammary progenitor cell, an embryonic kidney cell (e.g. HEK293 or HEK 293T cell), a fibroblast cell, a COS cell, a Chinese hamster ovary (CHO) cell, a murine myeloma cell like e.g. an 1X120, SP2/0 or YB2/0 cell, an NIH-3T3 cell, a non-mammary adult stem cell or derivatives thereof such as described in WO21067641, preferably mesenchymal stem cell or derivates thereof as described in WO21067641. Said insect cell is preferably derived from Spodoptera frugiperda like e.g. Sf9 or Sf21 cells, Bombyx mori, Mamestra brassicae, Trichoplusia ni like e.g. BTI-TN-5B1-4 cells or Drosophila melanogaster like e.g. Drosophila S2 cells. The latter protozoan cell preferably is a Leishmania tarentolae cell.

In an additional and/or alternative preferred embodiment, said saccharide of the invention has been produced, preferably in vitro and/or ex vivo, by a mammary epithelial cell, mammary myoepithelial cell and/or mammary progenitor cell, preferably wherein said cell is generated from non-mammary adult stem cells, more preferably wherein said cell is generated from mesenchymal stem cells. Such cells are well-known to the skilled person, it is in this regard referred to for example WO2021/067641 and WO2021/242866 (mammary epithelial cells derived from non-mammary adult stem cells, preferably from mesenchymal stem cells) and WO2021/142241 (mammary epithelial cells, mammary myoepithelial cells, mammary progenitor cells).

In another additional and/or alternative preferred embodiment, said saccharide of the invention has been produced, preferably in vitro and/or ex vivo, by a microorganism cell, preferably said microorganism is a bacterium, a yeast or a fungus, more preferably said microorganism is a bacterium or a yeast, even more preferably said microorganism is a bacterium, most preferably said microorganism is Escherichia coli.

In the context of the invention, it is also within the scope of the present invention that two or more different cells (preferably as defined herein), produce the saccharide according to the invention. It is however preferred that a single cell produces said saccharide, i.e. a single culture of said cell produces the saccharide of the invention.

In an additional and/or alternative preferred embodiment, said saccharide according to the invention is typically isolated from a microbial cultivation or fermentation, cell culture, enzymatic reaction or chemical reaction as described herein, resulting in a solution containing said saccharide. Such a solution can for example be obtained by a method comprising the steps of:

(a) cultivating at least one cell as defined herein, preferably a single cell, that is capable to produce said saccharide in a suitable cultivation medium to form a cultivation broth, preferably wherein said cell is metabolically engineered for the production of said saccharide or said mixture, and

(b) purifying said saccharide from the cultivation broth by:

(i) clarifying the cultivation broth, and

(ii) removing salts and/or medium components form said clarified cultivation broth, and/or

(iii) concentrating said saccharide in said clarified cultivation broth, thereby providing a solution comprising a purified saccharide.

In the context of the invention, "clarifying the cultivation broth" refers to the removal of suspended particulates and contaminants, particularly cells, cell components, insoluble metabolites and debris produced by culturing the cell according to the invention. Clarification is preferably one or more of centrifugation, flocculation, decantation and/or filtration.

In the context of the invention, "removing salts and/or medium components form said clarified cultivation broth" refers to removing substantially all the proteins, as well as peptides, amino acids, RNA and DNA and any endotoxins and glycolipids from said clarified cultivation broth, preferably after it has been. In this step, proteins and related impurities can be removed from said saccharide in a conventional manner. Preferably, proteins, salts, by-products, color, endotoxins and other related impurities are removed from said saccharide by ultrafiltration, nanofiltration, two-phase partitioning, reverse osmosis, microfiltration, activated charcoal or carbon treatment, treatment with non-ionic surfactants, enzymatic digestion, tangential flow high-performance filtration, tangential flow ultrafiltration, electrophoresis (e.g. using slabpolyacrylamide or sodium dodecyl sulphate-polyacrylamide gel electrophoresis (PAGE)), affinity chromatography (using affinity ligands including e.g. DEAE-Sepharose, poly-L-lysine and polymyxin-B, endotoxin-selective adsorber matrices), ion exchange chromatography (such as but not limited to cation exchange, anion exchange, mixed bed ion exchange, inside-out ligand attachment), hydrophobic interaction chromatography and/or gel filtration (i.e., size exclusion chromatography), particularly by chromatography, more particularly by ion exchange chromatography or hydrophobic interaction chromatography or ligand exchange chromatography. With the exception of size exclusion chromatography, proteins and related impurities are retained by a chromatography medium or a selected membrane, said saccharide remains in the said saccharide containing clarified cultivation broth.

Further purification of said saccharide may be accomplished, for example, by use of (activated) charcoal or carbon, nanofiltration, ultrafiltration, electrophoresis, enzymatic treatment or ion exchange to remove any remaining DNA, protein, LPS, endotoxins, or other impurity. Alcohols, such as ethanol, and aqueous alcohol mixtures can also be used. Another purification step is accomplished by crystallization, evaporation or precipitation of the product.

Several drying techniques are known to the skilled person which can be used to obtain powder from a solution containing said saccharide. Powder is preferably obtained by spray drying, freeze drying, spray freeze-drying, crystallization, lyophilization, band or belt drying, drum or roller drying, and/or agitated thin film drying, preferably by spray drying, drum or roller drying, or agitated thin film drying, more preferably by spray drying or agitated thin film drying, most preferably by spray drying, of a solution containing said saccharide. Preferably, said saccharide according to the invention constitutes > 70.0 %, preferably > 75.0 %, more preferably > 80.0, (w/w) of said powder. Preferably, said powder contains < 15 wt. %, preferably < 10 wt. %, more preferably < 9 wt. %, more preferably < 8 wt. %, more preferably < 7 wt. %, even more preferably < 5 wt. %, even more preferably < 4 wt. % of liquid, even more preferably < 3 wt. % of liquid, even more preferably < 2 wt. % of liquid, most preferably < 1 wt. %, preferably wherein said liquid is water.

In an additional and/or alternative embodiment, said saccharide according to the invention is linked, preferably chemically linked, to a carrier for delivery of said saccharide. Preferably, said carrier is a ceramide-based carrier or a polypeptide-based carrier, more preferably said carrier is a ceramide-based carrier. Preferably, said polypeptide-based carrier is epsilon-polylysine, alfa-polylysine, poly(aspartic acid), polyglutamic acid or polyornithine. These carriers are commercially available (e.g. Sigma-Aldrich, Carbosynth). Said ceramide-based carrier is preferably selected from a list consisting of dl8:l/16:0, tl8:0- 16:0, tl8:0-hl6:0, tl8:0-h22:0 and tl8:0-h24:0. These ceramide carriers are commercially available and well-known to the skilled person and are for example described in W02010/037785 which is incorporated by reference. dl8:l/16:0 is also known as C16 ceramide and N-palmitoylsphingosine and therefore interchangeable used herein. tl8:0-16:0 is also known as C16 phytoceramide and N-hexadecanoyl phytosphingosine and therefore interchangeable used herein. tl8:0-hl6:0, tl8:0-h22:0 and tl8:0-h24:0 are glycosylinositolphosphoceramides (GIPCs). As known to the skilled person, "d" and "t" refer to the hydroxylation state of the whole ceramide or long-chain base moiety (d is 2 groups, t is 3 groups), whereas "h" denotes a hydroxylation of the fatty acyl group.

Bacterial infection

In an embodiment of the first aspect of the invention, said saccharide comprising a fucose is for use in a method for preventing and/or treating a bacterial infection in a subject. Preferably, said bacterium is gram-positive. In other words, said saccharide comprising a fucose is preferably for use in a method for preventing and/or treating a gram-positive bacterial infection in a subject.

In a preferred embodiment, said bacterial infection is an infection by a Streptococcus species, preferably wherein said Streptococcus species is not Streptococcus pneumoniae or Streptococcus pyogenes, more preferably an infection by a Streptococcus species selected from a list consisting of Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae, Streptococcus phocae, Streptococcus parauberis, Streptococcus penaeicida, Streptococcus egui, Streptococcus suis and Streptococcus zooepidemicus, even more preferably an infection by a Streptococcus species selected from a list consisting of Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae, Streptococcus phocae, Streptococcus parauberis and Streptococcus penaeicida, even more preferably an infection by a Streptococcus species selected from a list consisting of Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae, Streptococcus phocae and Streptococcus parauberis, even more preferably an infection by a Streptococcus species selected from a list consisting of Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae and Streptococcus parauberis, most preferably an infection by Streptococcus agalactiae. Throughout the application and claims, unless specifically stated otherwise, the term "Streptococcus species" is preferably replaced with "Streptococcus sp." or "Streptococcus spp." and vice versa. The terms "Streptococcus agalactiae” and "group B Streptococcus" are used interchangeably herein as they refer to the same bacterial species as understood by the skilled person.

In an additional and/or alternative preferred embodiment, said Streptococcus species is not Streptococcus pneumoniae. In an additional and/or alternative preferred embodiment, said Streptococcus species is not Streptococcus pyogenes.

In an additional and/or alternative preferred embodiment, said bacterium is a Streptococcus species which is able to infect at least one subject as described in the section "subject". The Center for Food Security & Public Health published an overview of different Streptococcus species and subjects which can be infected by said Streptococcus species and which is incorporated by reference ("Zoonotic Streptococcosis", updated September 2020 and accessible on https://www.cfsph.iastate.edu/Factsheets/pdfs/streptococcosi s.pdf).

In an additional and/or alternative preferred embodiment, said bacterium is a Streptococcus species which is able to infect an aquatic animal, preferably a water breathing animal, more preferably a fish or shrimp.

In an additional and/or alternative preferred embodiment, said bacterium is a Streptococcus species which is able to infect a fish, preferably a marine fish. Preferably said fish is selected from a list consisting of tilapia, bass, trout, salmon, flounder, turbot, mackerel, mullet, tuna, eel, catfish, pomfret, grouper, rockfish, rabbitfish, barramundi, grunter and pompano, more preferably said fish is selected from a list consisting of tilapia, bass, trout, flounder, turbot, mackerel, mullet, tuna, eel, grunter, pompano, grunter and grouper, even more preferably said fish is selected from a list consisting of tilapia, bass, trout, flounder, turbot, mackerel, mullet, tuna, eel and catfish, even more preferably said fish is selected from a list consisting of tilapia, salmon and catfish. Mishra et al, 2018 describes streptococcosis in fish and provides an overview of different fish and the Streptococcosis species known to be able to infect these different fish (it is referred to Table 1 of Mishra et al, 2018, which is incorporated by reference). More preferably said bacterium is a Streptococcus species selected from a list consisting of: subject is tilapia: Streptococcus agalactiae, Streptococcus iniae and Streptococcus dysgalactiae; subject is bass: Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae and Streptococcus parauberis; subject is trout: Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae and Streptococcus parauberis; subject is salmon: Streptococcus phocae; subject is flounder: Streptococcus iniae, Streptococcus dysgalactiae and Streptococcus parauberis; subject is turbot: Streptococcus parauberis; subject is mackerel: Streptococcus agalactiae, Streptococcus iniae and Streptococcus dysgalactiae; subject is mullet: Streptococcus agalactiae and Streptococcus dysgalactiae; subject is tuna: Streptococcus dysgalactiae; subject is eel: Streptococcus agalactiae and Streptococcus iniae; subject is catfish: Streptococcus agalactiae, Streptococcus iniae and Streptococcus dysgalactiae; subject is pomfret: Streptococcus agalactiae and Streptococcus dysgalactiae; subject is grouper: Streptococcus agalactiae; subject is rockfish: Streptococcus agalactiae and Streptococcus iniae; subject is rabbitfish: Streptococcus iniae; subject is barramundi: Streptococcus iniae; subject is grunter: Streptococcus agalactiae; subject is pompano: Streptococcus agalactiae and Streptococcus dysgalactiae.

In an additional and/or alternative preferred embodiment, said bacterium is a Streptococcus species which is able to infect a crustacean. Preferably, said Streptococcus species is Streptococcus penaeicida. This species has been described by for example Morales-Covarrubias et al, 2018.

In an additional and/or alternative preferred embodiment, said bacterium is a Streptococcus species which is able to infect a farmed animal. Preferably, said farmed animal is selected from a list consisting of bovine, pig, horse, sheep, goat, camelid and poultry. More preferably, said farmed animal is a pig, bovine or horse. Even more preferably, said bacterium is a Streptococcus species selected from a list consisting of: subject is bovine: Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus parauberis; subject is a pig: Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus suis; subject is a horse: Streptococcus agalactiae, Streptococcus egui and Streptococcus zooepidemicus; subject is a sheep: Streptococcus dysgalactiae and Streptococcus zooepidemicus; subject is a goat: Streptococcus dysgalactiae, Streptococcus egui and Streptococcus zooepidemicus; subject is a camelid: Streptococcus agalactiae, Streptococcus equi and Streptococcus zooepidemicus; subject is poultry: Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus suis, Streptococcus equi and Streptococcus zooepidemicus.

In an additional and/or alternative preferred embodiment, said bacterium is a Streptococcus species which is able to infect a human. Preferably, said bacterium is selected from a list consisting of Streptococcus agalactiae, Streptococcus iniae and Streptococcus dysgalactiae.

In an additional and/or alternative preferred embodiment, said bacterium is a Streptococcus species which is able to infect a horse. Preferably, said bacterium is selected from a list consisting of Streptococcus agalactiae, Streptococcus equi and Streptococcus zooepidemicus.

In an additional and/or alternative preferred embodiment, said bacterium is a Streptococcus species which is able to infect a companion animal, preferably wherein said companion animal is a dog or a cat. Preferably, said bacterium is Streptococcus agalactiae or Streptococcus dysgalactiae.

In an additional and/or alternative preferred embodiment, said bacterium is a Streptococcus species which is able to infect a reptile. Preferably, said bacterium is Streptococcus agalactiae.

In an additional and/or alternative preferred embodiment, said saccharide comprising a fucose is for use in a method for preventing and/or treating streptococcosis. The latter is a bacterial disease caused by a Streptococcus species, preferably as described herein. Throughout the application, unless specifically stated otherwise, "for use in a method for preventing and/or treating a bacterial infection" is preferably replaced with "for use in a method for preventing and/or treating streptococcosis".

Subject

In an embodiment of the first aspect of the invention, said saccharide comprising a fucose is for use in a method for preventing and/or treating a bacterial infection in a subject. Preferably, said subject is a human or an animal, more preferably said subject is an animal. Throughout the description and claims, unless specifically stated otherwise, the term "subject" can be preferably replaced with "subject in the need thereof".

Preferably, said animal is selected from a list consisting of an aquatic animal, a horse, a farmed animal, a companion animal and a reptile, preferably wherein said animal is an aquatic animal or a farmed animal, more preferably wherein said animal is an aquatic animal. In a preferred embodiment, said farmed animal is selected from a list consisting of bovine, pig, horse, sheep, goat, camelid and poultry, preferably said farmed animal is bovine, pig or horse.

In an additional and/or alternative embodiment, said companion animal is a dog or cat.

In an additional and/or alternative embodiment, said aquatic animal is a water breathing animal, preferably wherein said water breathing animal is a fish or crustacean, preferably a fish, mor preferably a marine fish. Preferably, said crustacean is a shrimp.

In a more preferred embodiment, said fish is selected from a list consisting of tilapia, bass, trout, salmon, flounder, turbot, mackerel, mullet, tuna, eel, catfish, pomfret, grouper, rockfish, rabbitfish, barramundi, grunter and pompano, preferably said fish is selected from a list consisting of tilapia, bass, trout, flounder, turbot, mackerel, mullet, tuna, eel, grunter, pompano, grunter and grouper, more preferably said fish is selected from a list consisting of tilapia, bass, trout, flounder, turbot, mackerel, mullet, tuna, eel and catfish, even more preferably said fish is selected from a list consisting of tilapia, salmon and catfish.

In an additional and/or alternative more preferred embodiment, said fish is selected from a list consisting of tilapia, grunter, pompano and pomfret, preferably wherein said fish is tilapia.

As understood by the skilled person, a "farmed animal" refers to an animal that is reared in an agricultural setting in order to produce various commodities such as food (meat, organs, eggs, dairy products) and/or hair or wool. Further, a "companion animal" preferably refers to a domestic animal. Further, an "aquatic animal" is an animal that lives in water for most or all of its lifetime, said animal can be an invertebrate or a vertebrate. Further, a "water breathing animal" or "water breathing aquatic animal" refers to an aquatic animal which is able to breath under water.

In a preferred embodiment, said subject is an adult or a non-adult subject, preferably said subject is a non-adult subject.

In an additional and/or alternative preferred embodiment, said subject is a non-adult human, preferably an infant (age of 0-1 year) or a child of 1-9 years, more preferably an infant (age of 0-1 year) or a child of 1-6 years, even more preferably an infant (age of 0-1 year) or a child of 1-4 years.

In an additional and/or alternative preferred embodiment, said subject is a non-adult animal, preferably selected from a list consisting of a fry, fingerling, foal, calf, piglet, lamb, kid, cria, chick, puppy and kitten, more preferably a fry, fingerling, foal, calf or piglet, even more preferably a fry or fingerling, most preferably a fingerling. As the skilled person understands, a non-adult animal of a fish, crustacean, pig, bovine, horse, sheep, goat, camelid, poultry, dog, cat (as described earlier herein) is called a fry (young fish which is capable of feeding h im self )/f ingerl i ng (young fish which is capable of feeding himself and has developed scales and working fins, i.e. transition to a juvenile fish is complete), fry, piglet, calf, foal, lamb, kid, cria, chick, puppy, kitten, respectively. Throughout the application, unless specifically stated otherwise, a "non-adult animal" can be replaced with "juvenile animal" and vice versa.

Method for preventing and/or treating

In an embodiment of the first aspect of the invention, a saccharide according to the invention is for use in a method for preventing and/or treating a bacterial infection in a subject. In an additional and/or alternative embodiment, a saccharide according to the invention is for use in a method for preventing and/or treating streptococcosis.

In a preferred embodiment, said method according to the invention comprises administering an effective amount of said saccharide according to the invention to said subject. As understood by the skilled person, an "effective amount" is the amount of said saccharide which is required to confer a therapeutic effect on the subject as described in the present application. Effective amounts vary, as recognized by those skilled in the art, depending on the subject, route of administration, excipient usage among other known factors.

In an additional and/or alternative embodiment, said saccharide according to the invention is administered a daily dose of 0.01-150.0 mg, preferably 0.01-125.0 mg, more preferably 0.01-100.0 mg, even more preferably 0.01-80.0, even more preferably 0.01-70.0, even more preferably 0.01-60.0, most preferably 0.01-50.0 mg, per kg bodyweight of said subject. In the context of the present invention, an amount of a saccharide according to the invention expressed in a number of grams or milligrams per daily dose as used herein means that the amount of the saccharide is such that when administering the daily dosage to a subject, the subject will be administered with the number of grams or milligrams of the saccharide. In other words, if the daily dosage is for example 50 mg, then the subject receives in total 50 mg per day. This may be in one or more portions. So, if the daily dosage is 50 mg divided over 2 portions, then a single serving consists of 25 mg, a daily serving consists of 2 of such single servings.

In an additional and/or alternative embodiment, said saccharide according to the invention is administered at a daily dose of 0.1-70.0 mg, preferably 0.1-60.0 mg, more preferably 0.1-50.0 mg, even more preferably 0.5-50.0 mg, even more preferably 1.0-50.0 mg, even more preferably 2.5-50.0 mg, even more preferably 5.0-50.0 mg, even more preferably 10.0-50.0 mg, most preferably 10.0-40.0 mg, per kg bodyweight of said subject. This is for example particularly preferred if said subject is a fish or crustacean, preferably a non-adult fish or crustacean. In an additional and/or alternative embodiment, said saccharide according to the invention is administered at daily dose of 1.0-150.0 mg, preferably 1.0-125.0 mg, more preferably 5.0-125.0 mg, even more preferably 10.0-125.0 mg, even more preferably 10.0-100.0 mg, even more preferably 10.0-80.0 mg, even more preferably 15.0-80.0 mg, even more preferably 20.0-80.0 mg, even more preferably 25.0- 80.0 mg, even more preferably 30.0-80.0 mg, even more preferably 35.0-80.0 mg, even more preferably 40.0-80.0 mg, most preferably 45.0-80.0 mg, per kg bodyweight of said subject. This is for example particularly preferred if said subject is a pig, preferably a piglet.

In an additional and/or alternative embodiment, said saccharide according to the invention is administered at daily dose of 0.01-50.0 mg, preferably 0.01-30.0 mg, more preferably 0.01-25.0 mg, even more preferably 0.01-20.0 mg, even more preferably 0.01-15.0 mg, even more preferably 0.01-10.0 mg, even more preferably 0.05-10.0 mg, most preferably 0.1-10.0 mg, per kg bodyweight of said subject. This is for example particularly preferred if said subject is bovine, preferably a non-adult bovine.

In an additional and/or alternative embodiment, said saccharide according to the invention is administered at a daily dose of 0.0001-15.0 g, preferably 0.0001-10.0 g, more preferably 0.0001-5.0 g, even more preferably 0.0001-2.0 g, even more preferably 0.0001-1.0 g, even more preferably 0.0005-1.0 g, even more preferably 0.0005-0.5 g, even more preferably 0.0005-0.25 g, most preferably 0.0005-0.10 g-

In an additional and/or alternative embodiment, said saccharide according to the invention is administered at a daily dose of 0.0001-1.0 g, preferably 0.0001-0.50 g, more preferably 0.0001-0.10 g, even more preferably 0.0001-0.075 g, even more preferably 0.0001-0.050 g, even more preferably 0.0002-0.050 g, even more preferably 0.0005-0.050 g, most preferably 0.0005-0.025 g. This is for example particularly preferred if said subject is a fish or crustacean, preferably a non-adult fish or crustacean.

In an additional and/or alternative embodiment, said saccharide according to the invention is administered at a daily dose of 0.01-5.0 g, preferably 0.1-2.5 g, more preferably 0.1-1.5 g, even more preferably 0.1-1.2 g, even more preferably 0.2-1.2 g, even more preferably 0.3-1.2 g, even more preferably 0.4-1.2 g, most preferably 0.5-1.2 g. This is for example particularly preferred if said subject is a pig, preferably a piglet.

In an additional and/or alternative embodiment, said saccharide according to the invention is administered at daily dose of 0.01-1.0 g, preferably 0.01-0.75 g, more preferably 0.01-0.65 g, even more preferably 0.01-0.60 g, even more preferably 0.01-0.50 g, even more preferably 0.015-0.50 g, even more preferably 0.015-0.40 g, even more preferably 0.015-0.30 g, most preferably 0.015-0.20. This is for example particularly preferred if said subject is bovine, preferably a non-adult bovine.

In a more preferred embodiment, said saccharide according to the invention is administered to said subject at least once between birth (0 weeks) and 5 weeks of age, preferably between birth and 4 weeks of age, more preferably between birth and 3 weeks of age.

In an additional and/or alternative more preferred embodiment, said saccharide according to the invention is administered to said subject for 1-30, preferably 1-25, more preferably 1-20, even more preferably 1-15, even more preferably 1-10, even more preferably 1-7.5, even more preferably 1-5, even more preferably 1-4, most preferably 2-4, consecutive weeks. Preferably, said saccharide according to the invention is administered at least once a week (i.e. at least one daily dosage is administered in a week), more preferably at least once every 3 days, even more preferably at least once every 2 days, most preferably at least once daily (i.e. a daily dosage is administered every day).

In an additional and/or alternative more preferred embodiment, said saccharide according to the invention is administered to said subject for:

If subject is a fish: 1-6, preferably 1-5, more preferably 1-4, even more preferably 2-4, most preferably 3-4, consecutive weeks;

If subject is a crustacean: 1-6, preferably 1-5, more preferably 1-4, even more preferably 2-4, most preferably 3-4, consecutive weeks;

If subject is bovine: 1-36, preferably 1-32, more preferably 1-30, even more preferably 1-28, even more preferably 1-26, even more preferably 1-18, even more preferably 1-12, even more preferably 1-6, most preferably 1-4, consecutive weeks;

If subject is a pig: 1-5, preferably 1-4, more preferably 1-3, most preferably 2-3, consecutive weeks;

If subject is a horse: 1-12, preferably 1-8, more preferably 1-5, even more preferably 1-3, most preferably 1-2, consecutive weeks;

If subject is a sheep: 1-36, preferably 1-32, more preferably 1-30, even more preferably 1-28, even more preferably 1-26, even more preferably 1-18, even more preferably 1-12, even more preferably 1-6, most preferably 1-4, consecutive weeks;

If subject is a goat: 1-36, preferably 1-32, more preferably 1-30, even more preferably 1-28, even more preferably 1-26, even more preferably 1-18, even more preferably 1-12, even more preferably 1-6, most preferably 1-4, consecutive weeks;

If subject is a camelid: 1-12, preferably 1-10, more preferably 1-8, even more preferably 1-6, even more preferably 1-5, even more preferably 1-4, even more preferably 1-3, most preferably 1-2, consecutive weeks;

If subject is poultry: 1-5, preferably 1-4, more preferably 1-3, most preferably 2-3, consecutive weeks;

If subject is a dog: 1-5, preferably 1-4, more preferably 1-3, most preferably 2-3, consecutive weeks;

If subject is a cat: 1-5, preferably 1-4, more preferably 1-3, most preferably 2-3, consecutive weeks;

If subject is a reptile: 1-12, preferably 1-8, more preferably 1-5, even more preferably 1-3, most preferably 1-2 consecutive weeks;

If subject is a human: 1-12, preferably 1-10, more preferably 1-8, even more preferably 1-6, even more preferably 1-5, even more preferably 1-4, even more preferably 1-3, most preferably 1-2, consecutive weeks.

Preferably, said saccharide according to the invention is administered at least once a week (i.e. at least one daily dosage is administered in a week), more preferably at least once every 3 days, even more preferably at least once every 2 days, most preferably at least once daily (i.e. a daily dosage is administered every day).

The inventors have surprisingly found that a saccharide according to the invention is sufficient to obtain an effective therapeutic for preventing and/or treating bacterial disease such as streptococcosis. The presence of other (oligo)saccharides is not required to obtain said therapeutic effect. While the saccharide according to the invention can be administered in a composition including additional components such as saccharides, the present invention relates to the fact that only the saccharide according to the invention is required for achieving an efficient therapeutic effect as described herein.

In an even more preferred embodiment, said saccharide according to the invention is administered to said subject as part of a composition, wherein said saccharide is present at a higher amount or concentration than any mammalian milk oligosaccharide present in said composition. The skilled person will understand that such mammalian milk oligosaccharide(s) can be absent in said composition and that hence the amount or concentration of the saccharide according to the invention is inevitably higher. Further, if one or more mammalian milk oligosaccharides are present in said composition, than the amount or concentration of the saccharide according to the invention (which can be a mammalian milk oligosaccharide or not as defined earlier herein) is higher than said one or more mammalian milk oligosaccharides. In case said saccharide according to the invention is a mammalian milk oligosaccharide, then it is preferred that said saccharide according to the invention is administered to said subject as part of a composition, wherein said saccharide is present at a higher amount or concentration than any other mammalian milk oligosaccharide present in said composition. Preferably said composition is a pharmaceutical composition and/or a nutritional composition, more preferably a pharmaceutical composition and/or a nutritional composition as described later herein.

In an additional and/or alternative even more preferred embodiment, said saccharide according to the invention is administered to said subject in the absence of lacto-N-neotetraose and/or lacto-N-tetraose and/or sialyllactose, preferably in the absence of lacto-N-neotetraose, more preferably in the absence of lacto-N-neotretraose and lacto-N-tetraose, even more preferably in the absence of lacto-N-neotretraose, lacto-N-tetraose and sialyllactose. In the context of the present invention, the expression "X in the absence of Y" preferably means that the amount of Y constitutes < 10%, preferably < 5.0%, more preferably < 2.0%, even more preferably < 1.0%, most preferably < 0.5%, of the amount of X in a composition administered to said subject. More preferably, said expression means that Y is not present in a composition comprising X.

In an additional and/or alternative even more preferred embodiment, said saccharide according to the invention is administered to said subject in the absence of LNFP I and/or LNFP V and/or LNnFP V and/or LNDFH II and/or LNnDFH, preferably in the absence of LNFP I, more preferably in the absence of LNFP I and LNFP V, even more preferably in the absence of LNFP I, LNFP V and LNnFP V, even more preferably in the absence of LNFP I, LNFP V, LNnFP V and LNDFH II, most preferably in the absence of LNFP I, LNFP V, LNnFP V, LNDFH II and LNnDFH.

In a second aspect, the invention provides a composition for use in a method for preventing and/or treating a bacterial infection in a subject, said composition comprises a saccharide comprising a fucose. In an additional and/or alternative embodiment, the invention provides a composition for use in a method for preventing and/or treating streptococcosis in a subject, said composition comprises a saccharide comprising a fucose.

In a preferred embodiment, said composition according to the invention comprises a saccharide as described in the section "saccharide" of the first aspect of the invention.

In an additional and/or alternative preferred embodiment, said bacterial infection is as described in the section "bacterial infection" of the first aspect of the invention.

In an additional and/or alternative preferred embodiment, said subject is as described in the section "subject" of the first aspect of the invention.

In an additional and/or alternative preferred embodiment, said method for preventing and/or treating is as described in the section "method for preventing and/or treating" of the first aspect of the invention. For example, a preferred embodiment of the second aspect of the invention relates to a composition for use in a method for preventing and/or treating a bacterial infection in a subject, said composition comprises a saccharide comprising a fucose, wherein said method comprises administering an effective amount of said saccharide to said subject.

In an additional and/or preferred embodiment, said saccharide of a composition according to the invention is present in said composition at 0.001-5.000 wt. %, preferably 0.001-2.500 wt. %, more preferably 0.001-1.000 wt. %, even more preferably 0.001-0.750 wt. %, even more preferably 0.001-0.500 wt. %, even more preferably 0.002-0.500 wt. %, even more preferably 0.002-0.400 wt. %, most preferably 0.002-0.250 wt. %. In an alternative preferred embodiment, said saccharide of a composition according to the invention is present in said composition at 0.001-2.500 wt. %, preferably 0.001-1.000 wt. %, more preferably 0.010-1.000 wt. %, even more preferably 0.025-1.000 wt. %, even more preferably 0.025-0.750 wt. %, even more preferably 0.025-0.500 wt. %, even more preferably 0.025-0.250 wt. %, even more preferably 0.025-0.100 wt. %, most preferably 0.025-0.075 wt. %. This is for example particularly preferred if said subject is a fish or crustacean, preferably a non-adult fish or crustacean. In an alternative preferred embodiment, said saccharide of a composition according to the invention is present in said composition at 0.010-1.000 wt. %, preferably 0.010-0.750 wt. %, more preferably 0.010-0.500 wt. %, even more preferably 0.010-0.400 wt. %, even more preferably 0.050-0.400 wt. %, even more preferably 0.100-0.400 wt. %, most preferably 0.100-0.300 wt. % This is for example particularly preferred if said subject is a pig, preferably a piglet. In an alternative preferred embodiment, said saccharide of a composition according to the invention is present in said composition at 0.001-0.250 wt. %, preferably 0.001-0.200 wt. %, more preferably 0.001-0.100 wt. %, even more preferably 0.001-0.020 wt. %, even more preferably 0.001-0.010 wt. %, even more preferably 0.002-0.010 wt. %, even more preferably 0.003-0.010 wt. %, most preferably 0.003-0.0075 wt. %. This is for example particularly preferred if said subject is bovine, preferably a nonadult bovine.

In a more preferred embodiment, said saccharide of a composition according to the invention is present at a higher amount or concentration than any mammalian milk oligosaccharide present in said composition. The skilled person will understand that such mammalian milk oligosaccharide(s) can be absent in said composition and that hence the amount or concentration of the saccharide according to the invention is inevitably higher. Further, if one or more mammalian milk oligosaccharides are present in said composition, than the amount or concentration of the saccharide according to the invention (which can be a mammalian milk oligosaccharide or not as defined earlier herein) is higher than said one or more mammalian milk oligosaccharides. In case said saccharide according to the invention is a mammalian milk oligosaccharide, then it is preferred that said saccharide according to the invention is present at a higher amount or concentration than any other mammalian milk oligosaccharide present in said composition. In an additional and/or alternative more preferred embodiment, said composition: does not comprise lacto-N-neotetraose and/or lacto-N-tetraose and/or sialyllactose, preferably does not comprise lacto-N-neotetraose, more preferably does not comprise lacto-N-neotretraose and lacto-N-tetraose, even more preferably does not comprise lacto-N-neotretraose, lacto-N- tetraose and sialyllactose; or comprises any one or more of lacto-N-neotetraose, lacto-N-tetraose and sialyllactose, but at an amount in the composition which is at least 5, preferably 10, more preferably 15, even more preferably 20, even more preferably 25, even more preferably 50, times lower than the amount of said saccharide in the composition.

In an additional and/or alternative more preferred embodiment, said composition: does not comprise LNFP I and/or LNFP V and/or LNnFP V and/or LNDFH II and/or LNnDFH, preferably does not comprise LNFP I, more preferably does not comprise LNFP I and LNFP V, even more preferably does not comprise LNFP I, LNFP V and LNnFP V, even more preferably does not comprise LNFP I, LNFP V, LNnFP V and LNDFH II, most preferably does not comprise LNFP I, LNFP V, LNnFP V, LNDFH II and LNnDFH; or comprises any one or more of LNFP I, LNFP V, LNnFP V, LNDFH II and LNnDFH, but at an amount in the composition which is at least 5, preferably 10, more preferably 15, even more preferably 20, even more preferably 25, even more preferably 50, times lower than the amount of said saccharide in the composition.

In an additional and/or alternative more preferred embodiment, said saccharide is provided as a powder in a composition according to the invention. Preferably, said powder is as defined in the section "saccharide" of the first aspect.

In an even more preferred embodiment, said composition is a pharmaceutical composition, optionally further comprising a pharmaceutically acceptable carrier, filler, preservative, solubilizer, diluent, excipient, salt, adjuvant and/or solvent.

In an additional and/or alternative even more preferred embodiment, said composition is a nutritional composition, optionally further comprising a feed ingredient and/or a food ingredient, wherein said feed/food ingredient is preferably chosen from the list consisting of: a lipid, preferably one or more selected from the list consisting of an oil, fat, ester, monoglyceride, diglyceride, triglyceride and free fatty acid; a vitamin, preferably one or more selected from the list consisting of vitamin A, vitamin B, vitamin C, vitamin D, vitamin E and vitamin H, or a derivate thereof; an amino acid compound; a trace element; a mineral, preferably one or more selected from the list consisting of calcium, phosphorus, magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, selenium and chloride; an antioxidant; a prebiotic agent, preferably one or more selected from the list consisting of GOS (galactooligosaccharide), FOS (fructo-oligosaccharide), inulin and resistant starch; a carbohydrate; an antimicrobial agent; and/or a protein.

Generally, any source of protein may be used so long as it is suitable for nutritional compositions and is otherwise compatible with any other selected ingredients or features in the nutritional composition. Nonlimiting examples of suitable proteins (and sources thereof) suitable for use in the nutritional composition according to the invention include, but are not limited to, intact, hydrolyzed, or partially hydrolyzed protein, which may be derived from any known or otherwise suitable source such as milk (e.g., casein, whey), animal (e.g., meat, fish), cereal (e.g., rice, corn, wheat), vegetable (e.g., soy, pea, potato, bean), and combinations thereof. The protein may also include a mixture of amino acids (often described as free amino acids) known for use in nutritional products or a combination of such amino acids with the intact, hydrolyzed, or partially hydrolyzed proteins described herein. The amino acids may be naturally occurring or synthetic amino acids. More particular examples of suitable protein (or sources thereof) used in a nutritional composition according to the invention include, but are not limited to, whole cow's milk, partially or completely defatted milk, milk protein concentrates, milk protein isolates, nonfat dry milk, condensed skim milk, whey protein concentrates, whey protein isolates, acid caseins, sodium caseinates, calcium caseinates, potassium caseinates, legume protein, soy protein concentrates, soy protein isolates, pea protein concentrates, pea protein isolates, collagen proteins, potato proteins, rice proteins, wheat proteins, canola proteins, quinoa, insect proteins, earthworm proteins, fungal (e.g., mushroom) proteins, hydrolyzed yeast, gelatin, bovine colostrum, human colostrum, glycol macropeptides, mycoproteins, proteins expressed by microorganisms (e.g., bacteria and algae), and combinations thereof. A nutritional composition according to the invention may include any individual source of protein or combination of the various sources of protein listed above. In addition, the proteins for use herein can also include, or be entirely or partially replaced by, free amino acids known for use in nutritional products, non-limiting examples of which include L-tryptophan, L-glutamine, L-tyrosine, L-methionine, L-cysteine, taurine, L- arginine, L-carnitine, and combinations thereof.

The carbohydrate or source of carbohydrate suitable for use in a nutritional composition according to the invention may be simple, complex, or variations or combinations thereof. Generally, the carbohydrate may include any carbohydrate or carbohydrate source that is suitable for use nutritional compositions and is otherwise compatible with any other selected ingredients or features in the nutritional composition. Non-limiting examples of carbohydrates suitable for use in the nutritional composition according to the invention, but are not limited to, polydextrose, maltodextrin; hydrolyzed or modified starch or cornstarch; glucose polymers; corn syrup; corn syrup solids; sucrose; glucose; fructose; lactose; high fructose corn syrup; honey; sugar alcohols (e.g., maltitol, erythritol, sorbitol); isomaltulose; sucromalt; pullulan; potato starch; and other slowly-digested carbohydrates; dietary fibers including, but not limited to, fructooligosaccharides (FOS), galactooligosaccharides (GOS), oat fiber, soy fiber, gum arabic, sodium carboxymethylcellulose, methylcellulose, guar gum, gellan gum, locust bean gum, konjac flour, hydroxypropyl methylcellulose, tragacanth gum, karaya gum, gum acacia, chitosan, arabinogalactans, glucomannan, xanthan gum, alginate, pectin, low methoxy pectin, high methoxy pectin, cereal beta-glucans (e.g., oat beta-glucan, barley beta-glucan), carrageenan and psyllium, soluble and insoluble fibers derived from fruits or vegetables; other resistant starches; and combinations thereof. A nutritional composition according to the invention may include any individual source of carbohydrate or combination of the various sources of carbohydrate listed above.

The fat or source of fat suitable for use in a nutritional composition according to the invention may be derived from various sources including, but not limited to, plants, animals, and combinations thereof. Generally, the fat may include any fat or fat source that is suitable for use in a nutritional composition according to the invention and is otherwise compatible with any other selected ingredients or features in the nutritional composition. Non-limiting examples of suitable fat (or sources thereof) for use in a nutritional composition according to the invention include coconut oil, fractionated coconut oil, soy oil, high oleic soy oil, corn oil, olive oil, safflower oil, high oleic safflower oil, medium chain triglyceride oil (MCT oil), high gamma linolenic (GLA) safflower oil, sunflower oil, high oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil, high oleic canola oil, marine oils, fish oils, algal oils, borage oil, cottonseed oil, fungal oils, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), arachidonic acid (ARA), conjugated linoleic acid (CLA), alpha-linolenic acid, rice bran oil, wheat bran oil, interesterified oils, transesterified oils, structured lipids, and combinations thereof. Generally, the fats used in a nutritional composition for formulating infant formulas and pediatric formulas provide fatty acids needed both as an energy source and for the healthy development of the infant, toddler, or child. These fats typically comprise triglycerides, although the fats may also comprise diglycerides, monoglycerides, and free fatty acids. Fatty acids provided by the fats in the nutritional composition include, but are not limited to, capric acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, alphalinolenic acid, ARA, EPA, and DHA. The nutritional composition can include any individual source of fat or combination of the various sources of fat listed above. Preferably, the fat is a mixture of vegetable fat and milk fat such as obtained from milk from a mammal like cow, sheep, goat, mare, or camel. More preferably, wherein the milk fat is bovine milk fat. Mixtures of different types of fat are preferred because they help to provide different fatty acids and better resemble the type of linkage between the glycerol moiety and the fatty acid moiety in the fat, when compared to human mother's milk.

When one or more prebiotics is/are present, then it is preferred that the weight ratio between the saccharide according to the invention and said prebiotic(s) is in the range of from 0.5:10 to 10:0.5.

In a preferred embodiment, said pharmaceutical or nutritional composition according to the invention comprises one or more probiotics for its beneficial effect on the subject's gut microbiome. Examples of probiotics include Bifidobacterium, Lactobacillus and Saccharomyces boulardii.

In a more preferred embodiment, said nutritional composition is a synthetic nutritional composition. In the context of the present invention, a "synthetic composition" or a "synthetic nutritional composition" refers to a composition which is artificially prepared and preferably refers to a composition comprising at least one component that is produced ex vivo, either chemically and/or biologically, e.g. by means of chemical reaction, enzymatic reaction or recombinantly, or purified by humans. It is preferred that a synthetic nutritional composition of the invention is not identical with a naturally occurring composition.

In an additional and/or alternative more preferred embodiment, said nutritional composition is a food product, preferably wherein said food product is selected from a list consisting of dairy product, bar, liquid product, savory snack, savory biscuit, bakery product, pasta and food supplement, more preferably wherein said food product is selected from a list consisting of dairy product, liquid product and food supplement.

In an additional and/or alternative more preferred embodiment, said nutritional composition is selected from a list consisting of infant formula, baby food, infant cereal composition, growing-up milk, milk replacer, creep feed, pet food, dry feed, frozen feed, pre weaning feed, weaning feed and post weaning feed.

In an alternative more preferred embodiment, said nutritional composition is selected from a list consisting of dry feed, frozen feed, milk replacer, creep feed, pet food, prestarter diet, pre weaning feed, weaning feed and post weaning feed, preferably selected from a list consisting of dry feed, frozen feed, milk replacer, creep feed and pet food, more preferably wherein said nutritional composition is dry feed, frozen feed, milk replacer or creep feed, even more preferably wherein said nutritional composition is dry feed or frozen feed, most preferably wherein said nutritional composition is dry feed.

In the context of the present invention, an "infant formula" refers to to a nutritional composition that has the proper balance of macronutrients, micro-nutrients, and calories to provide sole or supplemental nourishment for and generally maintain or improve the health of infants, toddlers, or both. Infant formulas preferably comprise nutrients in accordance with the relevant infant formula guidelines for the targeted consumer or user population, an example of which would be the Infant Formula Act, 21 U.S.C. Section 350(a). Another example with guidelines for nutrients of an infant formula, in particular for a person of 0-12 months of age and for children up to 36 months old, may be found in the CODEX Alimentarius (CODEX STAN 72-1981), further referred to as the CODEX). Nutritional compositions for infants are commonly referred to as infant formula. When used as infant formula, the nutritional composition according to the invention should contain the ingredients in the amounts as prescribed by the CODEX and, if needed, as prescribed by additional regulations of individual countries. The protein component is typically present in an amount of from 5% to 35% by weight of the infant formula (i.e., the dry weight), including from 10% to 30%, from 10% to 25%, from 15% to 25%, from 20% to 30%, from 15% to 20%, and also including from 10% to 16% by weight of the infant formula (i.e., the dry weight). The carbohydrate component is typically present in an amount of from 40% to 75% by weight of the infant formula (i.e., the dry weight), including from 45% to 75%, from 45% to 70%, from 50% to 70%, from 50% to 65%, from 50% to 60%, from 60% to 75%, from 55% to 65%, and also including from 65% to 70% by weight of the infant formula (i.e., the dry weight). The fat component is typically present in an amount of from 10% to 40% by weight of the infant formula (i.e., the dry weight), including from 15% to 40%, from 20% to 35%, from 20% to 30%, from 25% to 35%, and also including from 25% to 30% by weight of the infant formula (i.e., the dry weight).

In the context of the present invention, "growing-up milk" refers to a milk-based beverage adapted for the specific nutritional needs of young children.

In the context of the invention, the term "weaning" or "weaning period" refers to the period during which the mother's milk is substituted by other food in the diet.

In the context of the invention, the term "creep feed" refers to a nutritional composition which is used to supplement a pre-weaned subject with a solid diet while said subject is suckling. For example, a piglet receives creep feed to ease the transition from sow's milk to solid pig starter feed. The creep feed stimulates the digestive system of the piglet to produce e.g. amylase which digests carbohydrates in dry feed. If a pig is better able to digest dry feed, he can start eating quickly post-weaning for better performance.

In the context of the invention, the term "milk replacer" refers to a nutritional composition which serves as a substitute for mother's milk. A milk replacer comprises milk proteins, fat, carbohydrates, vitamins and minerals. Preferably, a milk replacer comprises 20 wt. % to 30 wt. % water, 18 wt. % to 24 wt. % of at least one protein, 15 wt. % to 28 wt. % (preferably 20 wt. % to 25 wt. %) of at least one fat and lactose at < 50 wt. %.

In an even more preferred embodiment, said nutritional composition is for feeding a fish or a crustacean, more preferably a fish. Preferably, said nutritional composition is a dry feed or frozen feed, more preferably a dry feed. As the skilled person is aware, such a feed can be pellet feed, flake feed or powdered feed. A nutritional composition for feeding a fish or a crustacean, preferably a fish, comprises proteins, fat, carbohydrates, vitamins and minerals. Preferably, said nutritional composition for feeding a fish or a crustacean, preferably a fish, comprises 10 wt. % to 20 wt. % fat and 15 wt. % to 35 wt. % carbohydrates. Preferably, said vitamin is any one or more of vitamin A, Bl, B2, B3, B5, B6, B12, biotin, C, choline, D3, E, folacin, inositol and K. Preferably, said mineral is any one or more of calcium, phosphorus, sodium, magnesium, iron, iodine, chloride, copper, potassium, sulfur and zinc, more preferably said mineral is calcium and/or phosphorus. Optionally, a binding agent for providing water stability to the feed is present. Preferably, said binding agent constitutes < 1.0 wt. % of the nutritional composition. Examples of suitable binding agents comprise guargum and carboxy methyl cellulose. Optionally, a preservative is present. Preferably, said preservative is an antioxidant and/or an anti-microbial. More preferably, said preservative is an antioxidant, preferably selected from a list consisting of vitamin E, butylated hydroxyanisole, butylated hydroxytoluene and ethoxyquine. More preferably, said preservative is an antimicrobial, preferably selected from a list consisting of sodium, benzoic and sorbic acid. It is preferred that said preservative constitutes < 0.1 wt. % of the nutritional composition. Optionally, an attractive is present. Its function is to make it more palatable. It is preferred that said attract constitutes < 5.0 wt/ % of the nutritional composition. Common sources of attractants are hydrolysates and condensed fish solubles. Optionally, a color enhancer is present. Its function is to obtain a vibrant color. Typically, carotenoids are added as color enhancer (e.g. astaxanthin and canthaxanthin are used for salmon feed). In the context of the invention, "pellet feed" refers to feed that is obtained by grinding up the feed components, extruding it with heat and pressure before producing the pellets in the desired size. Some pellet feed are designed to float on water, while others are designed to sink. "Flake feed" refers to feed that will float for a certain amount of time before slowly sinking to the bottom. "Powdered feed" refers to feed that is either directly fed or is mixed in water before feeding it. "Powdered feed" is often referred to as "fry feed".

Method for preventing and/or treating a parasite infection

In a third aspect, the invention provides a method for preventing and/or treating a bacterial infection in a subject as defined in the previous sections. The method comprises administering a saccharide comprising a fucose (as described in the section "saccharide" of the first aspect) or a composition (as described in the second aspect) of the invention to said subject.

In an additional and/or alternative preferred embodiment, the invention provides a method for preventing and/or treating streptococcosis. The latter is a bacterial disease caused by a Streptococcus species, preferably as described herein (it is referred to section "bacterial infection" of the first aspect). Throughout the application, unless specifically stated otherwise, "method for preventing and/or treating a bacterial infection" is preferably replaced with "method for preventing and/or treating streptococcosis".

Use

In a fourth aspect, the invention provides the use of a saccharide comprising a fucose (as described in the section "saccharide" of the first aspect) or a composition (as described in the second aspect) according to the invention for the manufacture of a medicament for preventing and/or treating a bacterial infection in a subject as defined in the previous sections.

In an additional and/or alternative preferred embodiment, the invention provides said use for the manufacture of a medicament for preventing and/or treating streptococcosis. The latter is a bacterial disease caused by a Streptococcus species, preferably as described herein (it is referred to section "bacterial infection" of the first aspect). Throughout the application, unless specifically stated otherwise, "manufacture of a medicament for preventing and/or treating a bacterial infection" is preferably replaced with "manufacture of a medicament for preventing and/or treating streptococcosis".

Specific embodiments

The present invention preferably relates to the following specific embodiments:

1. A saccharide comprising a fucose for use in a method for preventing and/or treating a bacterial infection in a subject, preferably wherein said bacterium is gram-positive.

2. A saccharide for use according to embodiment 1, wherein said bacterial infection is an infection by a Streptococcus species, preferably wherein said Streptococcus species is not Streptococcus pneumoniae or Streptococcus pyogenes, more preferably an infection by a Streptococcus species selected from a list consisting of Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae, Streptococcus phocae, Streptococcus parauberis, Streptococcus penaeicida, Streptococcus egui, Streptococcus suis and Streptococcus zooepidemicus, even more preferably an infection by a Streptococcus species selected from a list consisting of Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae, Streptococcus phocae, Streptococcus parauberis and Streptococcus penaeicida, even more preferably an infection by a Streptococcus species selected from a list consisting of Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae, Streptococcus phocae and Streptococcus parauberis, even more preferably an infection by a Streptococcus species selected from a list consisting of Streptococcus agalactiae, Streptococcus iniae, Streptococcus dysgalactiae and Streptococcus parauberis, most preferably an infection by Streptococcus agalactiae.

3. A saccharide for use according to embodiment 1 or 2, wherein said saccharide is for use in a method for preventing and/or treating streptococcosis. 4. A saccharide for use according to any one of embodiments 1 to 3, wherein said saccharide is a disaccharide or an oligosaccharide, preferably said saccharide is an oligosaccharide.

5. A saccharide for use according to embodiment 4, wherein said oligosaccharide consists of 3-9, preferably 3-8, more preferably 3-7, even more preferably 3-6, even more preferably 3-5, most preferably 3 or 4, monosaccharides.

6. A saccharide for use according to any one of embodiments 1 to 5, wherein said saccharide is a neutral saccharide.

7. A saccharide for use according to any one of embodiments 4 to 6, wherein said saccharide is a mammalian milk oligosaccharide (MMO), preferably a human milk oligosaccharide (HMO).

8. A saccharide for use according to any one of embodiments 1 to 7, wherein said fucose is linked to a monosaccharide in an alpha-1,2-, alpha-1,3- or alpha-1, 4-linkage, preferably an alpha-1,2- or an alpha- 1,3-linkage, more preferably an alpha-1, 3-linkage, and wherein said monosaccharide is preferably selected from glucose, N-acetylglucosamine and galactose, more preferably said monosaccharide is glucose or N-acetylglucosamine, even more preferably said monosaccharide is glucose.

9. A saccharide for use according to any one of embodiments 4 to 8, wherein said saccharide comprises a lactose, a lacto-N-biose (LNB) or N-acetyllactosamine (LacNAc) at its reducing end, preferably said saccharide comprises lactose or LacNAc at its reducing end, more preferably said saccharide comprises lactose at is reducing end.

10. A saccharide for use according to any one of embodiments 4 to 9, wherein said saccharide is selected from a list consisting of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3-FL), difucosyllactose (diFL), 2'- fucosyl-N-acetyllactosamine (2'FlacNAc), difucosyl-N-acetyllactosamine (diFLacNAc), 3-fucosyl-N- acetyllactosamine (3FlacNAc), 2'-fucosyllacto-N-biose (2'FLNB), 4-fucosyllacto-N-biose (4FLNB), difucosyllacto-N-biose (diFLNB), lacto-N-fucopentaose I (LNFP I), blood group A antigen hexaose type 1 (GalNAc-LNFP I), blood group B antigen hexaose type 1 (Gal-LNFP I), lacto-N-fucopentaose II (LNFP

II), lacto-N-fucopentaose III (LNFP III), lacto-N-fucopentaose V (LNFP V), lacto-N-difucohexaose I (LNDFH I), lacto-N-difucohexaose II (LNDFH II), lewis b-lewis x, monofucosyllacto-N-hexaose III (MFLNH

III), difucosyllacto-N-hexaose (a) (DFLNH (a)), difucosyllacto-N-hexaose (DFLNH), trifucosyllacto-N- hexaose (TFLNH), lacto-N-neofucopentaose I (LNnFP I), lacto-N-neofucopentaose V (LNnFP V, LNFP VI), and lacto-N-neodifucohexaose (LNnDFH), preferably selected from a list consisting of 2'FL, 3-FL, diFL, 2'FlacNAc, diFLacNAc, 3FlacNAc, 2'FLNB, 4FLNB, diFLNB, LNFP II, LNFP III, LNFP V, LNDFH I, lewis fa- lewis x, MFLNH III, DFLNH (a), DFLNH, TFLNH and LNnFP I.

11. A saccharide for use according to any one of embodiments 4 to 9, wherein said saccharide is selected from a list consisting of 3-FL, diFL, LNFP III, LNFP V, LNDFH II, lewis b-lewis x, MFLNH III, DFLNH (a), DFLNH, TFLNH, LNnFP V, LNnDFH, 3FlacNAc and diFLacNAc, preferably selected from a list consisting of 3-FL, diFL, LNFP V, LNDFH II, lewis b-lewis x, LNnFP V, LNnDFH, 3FlacNAc and diFLacNAc, more preferably selected from a list consisting of 3-FL, diFL, LNDFH II, lewis b-lewis x, LNnFP V, LNnDFH, 3FlacNAc and diFLacNAc, even more preferably selected from a list consisting of 3-FL, di FL, 3FlacNAc and diFLacNAc, even more preferably wherein said saccharide is 3-FL or 3FlacNAc, even more preferably wherein said oligosaccharide is 3-FL or di FL, most preferably wherein said saccharide is 3- FL.

12. A saccharide for use according to any one of embodiments 4 to 9, wherein said oligosaccharide is selected from a list consisting of 3-FL, 3FlacNAc, LNFP-III, MFLNH III, DFLNH (a), DFLNH and TFLNH, preferably selected from a list consisting of LNFP-III, MFLNH III, DFLNH (a), DFLNH and TFLNH, more preferably wherein said saccharide is LNFP-III.

13. A saccharide for use according to any one of embodiments 1 to 12, wherein said saccharide has been produced, preferably in vitro and/or ex vivo, by a cell, preferably a single cell, wherein said cell is preferably chosen from the list consisting of a microorganism, a plant cell, an animal cell or a protozoan cell.

14. A saccharide for use according to any one of embodiments 1 to 13, wherein said saccharide is linked, preferably chemically linked, to a carrier for delivery of said saccharide.

15. A saccharide for use according to embodiment 14, wherein said carrier is a ceramide-based carrier or a polypeptide-based carrier, preferably a ceramide-based carrier, wherein said ceramide-based carrier is preferably selected from a list consisting of dl8:l/16:0, tl8:0-16:0, tl8:0-hl6:0, tl8:0-h22:0 and tl8:0-h24:0.

16. A saccharide for use according to any one of embodiments 1 to 15, wherein said subject is a human or an animal, preferably said subject is an animal.

17. A saccharide for use according to embodiment 16, wherein said animal is selected from a list consisting of an aquatic animal, a horse, a farmed animal, a companion animal and a reptile, preferably wherein said animal is an aquatic animal or a farmed animal, more preferably wherein said animal is an aquatic animal.

18. A saccharide for use according to embodiment 17, wherein said farmed animal is selected from a list consisting of bovine, pig, horse, sheep, goat, camelid and poultry, and preferably wherein said companion animal is a dog or cat.

19. A saccharide for use according to embodiment 17 or 18, wherein said aquatic animal is a water breathing animal, preferably wherein said water breathing animal is a fish or crustacean, preferably a fish.

20. A saccharide for use according to embodiment 19, wherein said crustacean is a shrimp and/or wherein said fish is a marine fish.

21. A saccharide for use according to embodiment 19 or 20, wherein said fish is selected from a list consisting of tilapia, bass, trout, salmon, flounder, turbot, mackerel, mullet, tuna, eel, catfish, pomfret, grouper, rockfish, rabbitfish, barramundi, grunter and pompano, preferably said fish is selected from a list consisting of tilapia, bass, trout, flounder, turbot, mackerel, mullet, tuna, eel, grunter, pompano, grunter and grouper, more preferably said fish is selected from a list consisting of tilapia, bass, trout, flounder, turbot, mackerel, mullet, tuna, eel and catfish, even more preferably said fish is selected from a list consisting of tilapia, salmon and catfish.

22. A saccharide for use according to embodiment 19 or 20, wherein said fish is selected from a list consisting of tilapia, grunter, pompano and pomfret, preferably wherein said fish is tilapia.

23. A saccharide for use according to any one of embodiments 1 to 22, wherein said subject is an adult or a non-adult subject, preferably a non-adult subject.

24. A saccharide for use according to embodiment 23, wherein said subject is a non-adult human, preferably an infant (age of 0-1 year) or a child of 1-9 years, more preferably an infant (age of 0-1 year) or a child of 1-6 years, even more preferably an infant (age of 0-1 year) or a child of 1-4 years.

25. A saccharide for use according to embodiment 23, wherein said subject is a non-adult animal, preferably selected from a list consisting of a fry, fingerling, foal, calf, piglet, lamb, kid, cria, chick, puppy and kitten, more preferably a fry, fingerling, foal, calf or piglet, even more preferably a fry or fingerling, most preferably a fingerling.

26. A saccharide for use according to any one of embodiments 1 to 25, wherein said method comprises administering an effective amount of said saccharide to said subject.

27. A saccharide for use according to any one of embodiments 1 to 26, wherein said saccharide is administered at a daily dose of 0.01-150.0 mg, preferably 0.01-125.0 mg, more preferably 0.01-100.0 mg, even more preferably 0.01-80.0, even more preferably 0.01-70.0, even more preferably 0.01-60.0, most preferably 0.01-50.0 mg, per kg bodyweight of said subject.

28. A saccharide for use according to any one of embodiments 1 to 26, wherein said saccharide is administered at a daily dose of 0.1-70.0 mg, preferably 0.1-60.0 mg, more preferably 0.1-50.0 mg, even more preferably 0.5-50.0 mg, even more preferably 1.0-50.0 mg, even more preferably 2.5-50.0 mg, even more preferably 5.0-50.0 mg, even more preferably 10.0-50.0 mg, most preferably 10.0-40.0 mg, per kg bodyweight of said subject.

29. A saccharide for use according to any one of embodiments 1 to 26, wherein said saccharide is administered at a daily dose of 1.0-150.0 mg, preferably 1.0-125.0 mg, more preferably 5.0-125.0 mg, even more preferably 10.0-125.0 mg, even more preferably 10.0-100.0 mg, even more preferably 10.0-80.0 mg, even more preferably 15.0-80.0 mg, even more preferably 20.0-80.0 mg, even more preferably 25.0-80.0 mg, even more preferably 30.0-80.0 mg, even more preferably 35.0-80.0 mg, even more preferably 40.0-80.0 mg, most preferably 45.0-80.0 mg, per kg bodyweight of said subject.

30. A saccharide for use according to any one of embodiments 1 to 26, wherein said saccharide is administered at daily dose of 0.01-50.0 mg, preferably 0.01-30.0 mg, more preferably 0.01-25.0 mg, even more preferably 0.01-20.0 mg, even more preferably 0.01-15.0 mg, even more preferably 0.01- 10.0 mg, even more preferably 0.05-10.0 mg, most preferably 0.1-10.0 mg, per kg bodyweight of said subject. 31. A saccharide for use according to any one of embodiments 1 to 30, wherein said saccharide is administered at a daily dose of 0.0001-15.0 g, preferably 0.0001-10.0 g, more preferably 0.0001-5.0 g, even more preferably 0.0001-2.0 g, even more preferably 0.0001-1.0 g, even more preferably 0.0005- 1.0 g, even more preferably 0.0005-0.5 g, even more preferably 0.0005-0.25 g, most preferably 0.0005- 0.10 g.

32. A saccharide for use according to any one of embodiments 1 to 30, wherein said saccharide is administered at a daily dose of 0.0001-1.0 g, preferably 0.0001-0.50 g, more preferably 0.0001-0.10 g, even more preferably 0.0001-0.075 g, even more preferably 0.0001-0.050 g, even more preferably 0.0002-0.050 g, even more preferably 0.0005-0.050 g, most preferably 0.0005-0.025 g.

33. A saccharide for use according to any one of embodiments 1 to 30, wherein said saccharide is administered at a daily dose of 0.01-5.0 g, preferably 0.1-2.5 g, more preferably 0.1-1.5 g, even more preferably 0.1-1.2 g, even more preferably 0.2-1.2 g, even more preferably 0.3-1.2 g, even more preferably 0.4-1.2 g, most preferably 0.5-1.2 g.

34. A saccharide for use according to any one of embodiments 1 to 30, wherein said saccharide is administered at a daily dose of 0.01-1.0 g, preferably 0.01-0.75 g, more preferably 0.01-0.65 g, even more preferably 0.01-0.60 g, even more preferably 0.01-0.50 g, even more preferably 0.015-0.50 g, even more preferably 0.015-0.40 g, even more preferably 0.015-0.30 g, most preferably 0.015-0.20.

35. A saccharide for use according to any one of embodiments 1 to 34, wherein said saccharide is administered to said subject at least once between birth (0 weeks) and 5 weeks of age, preferably between birth and 4 weeks of age, more preferably between birth and 3 weeks of age.

36. A saccharide for use according to any one of embodiments 1 to 35, wherein said saccharide is administered to said subject for 1-30, preferably 1-25, more preferably 1-20, even more preferably 1- 15, even more preferably 1-10, even more preferably 1-7.5, even more preferably 1-5, even more preferably 1-4, most preferably 2-4, consecutive weeks.

37. A saccharide for use according to any one of embodiments 1 to 36, wherein said subject is administered to said subject for:

If subject is a fish: 1-6, preferably 1-5, more preferably 1-4, even more preferably 2-4, most preferably 3-4, consecutive weeks;

If subject is a crustacean: 1-6, preferably 1-5, more preferably 1-4, even more preferably 2-4, most preferably 3-4, consecutive weeks;

If subject is bovine: 1-36, preferably 1-32, more preferably 1-30, even more preferably 1-28, even more preferably 1-26, even more preferably 1-18, even more preferably 1-12, even more preferably 1-6, most preferably 1-4, consecutive weeks;

If subject is a pig: 1-5, preferably 1-4, more preferably 1-3, most preferably 2-3, consecutive weeks;

If subject is a horse: 1-12, preferably 1-8, more preferably 1-5, even more preferably 1-3, most preferably 1-2, consecutive weeks;

If subject is a sheep: 1-36, preferably 1-32, more preferably 1-30, even more preferably 1-28, even more preferably 1-26, even more preferably 1-18, even more preferably 1-12, even more preferably 1-6, most preferably 1-4, consecutive weeks;

If subject is a goat: 1-36, preferably 1-32, more preferably 1-30, even more preferably 1-28, even more preferably 1-26, even more preferably 1-18, even more preferably 1-12, even more preferably 1-6, most preferably 1-4, consecutive weeks;

If subject is a camelid: 1-12, preferably 1-10, more preferably 1-8, even more preferably 1-6, even more preferably 1-5, even more preferably 1-4, even more preferably 1-3, most preferably 1-2 consecutive weeks;

If subject is poultry: 1-5, preferably 1-4, more preferably 1-3, most preferably 2-3, consecutive weeks;

If subject is a dog: 1-5, preferably 1-4, more preferably 1-3, most preferably 2-3, consecutive weeks;

If subject is a cat: 1-5, preferably 1-4, more preferably 1-3, most preferably 2-3, consecutive weeks;

If subject is a reptile: 1-12, preferably 1-8, more preferably 1-5, even more preferably 1-3, most preferably 1-2, consecutive weeks;

If subject is a human: 1-12, preferably 1-10, more preferably 1-8, even more preferably 1-6, even more preferably 1-5, even more preferably 1-4, even more preferably 1-3, most preferably 1-2 consecutive weeks. A saccharide for use according to any one of embodiments 1 to 37, wherein said saccharide is administered to said subject as part of a composition, wherein said saccharide is present at a higher amount than any mammalian milk oligosaccharide present in said composition, preferably wherein said composition is a pharmaceutical composition and/or a nutritional composition. A saccharide for use according to any one of embodiments 1 to 38, wherein said saccharide is administered to said subject in the absence of lacto-N-neotetraose and/or lacto-N-tetraose and/or sialyllactose, preferably in the absence of lacto-N-neotetraose, more preferably in the absence of lacto-N-neotretraose and lacto-N-tetraose, even more preferably in the absence of lacto-N- neotretraose, lacto-N-tetraose and sialyllactose. A saccharide for use according to any one of embodiments 1 to 38, wherein said saccharide is administered to said subject in the absence of LNFP I and/or LNFP V and/or LNnFP V and/or LNDFH II and/or LNnDFH, preferably in the absence of LNFP I, more preferably in the absence of LNFP I and LNFP V, even more preferably in the absence of LNFP I, LNFP V and LNnFP V, even more preferably in the absence of LNFP I, LNFP V, LNnFP V and LNDFH II, most preferably in the absence of LNFP I, LNFP V, LNnFP V, LNDFH II and LNnDFH. 41. A composition for use in a method for preventing and/or treating a bacterial infection in a subject, said composition comprises a saccharide comprising a fucose, preferably said saccharide is as defined in any one of embodiments 4 to 15, wherein said bacterium is preferably gram-positive, more preferably said bacterial infection is as defined in embodiment 2.

42. A composition for use according to embodiment 41, wherein said composition is for use in a method for preventing and/or treating streptococcosis.

43. A composition for use according to embodiment 41 or 42, wherein said subject is as defined in any one of embodiments 16 to 25.

44. A composition for use according to any one of embodiments 41 to 43, wherein said saccharide is present in said composition at 0.001-5.000 wt. %, preferably 0.001-2.500 wt. %, more preferably 0.001- 1.000 wt. %, even more preferably 0.001-0.750 wt. %, even more preferably 0.001-0.500 wt. %, even more preferably 0.002-0.500 wt. %, even more preferably 0.002-0.400 wt. %, most preferably 0.002- 0.250 wt. %.

45. A composition for use according to any one of embodiments 41 to 43, wherein said saccharide is present in said composition at 0.001-2.500 wt. %, preferably 0.001-1.000 wt. %, more preferably 0.010- 1.000 wt. %, even more preferably 0.025-1.000 wt. %, even more preferably 0.025-0.750 wt. %, even more preferably 0.025-0.500 wt. %, even more preferably 0.025-0.250 wt. %, even more preferably 0.025-0.100 wt. %, most preferably 0.025-0.075 wt. %.

46. A composition for use according to any one of claims 41 to 43, wherein said saccharide is present in said composition at 0.010-1.000 wt. %, preferably 0.010-0.750 wt. %, more preferably 0.010-0.500 wt. %, even more preferably 0.010-0.400 wt. %, even more preferably 0.050-0.400 wt. %, even more preferably 0.100-0.400 wt. %, most preferably 0.100-0.300 wt. %.

47. A composition for use for use according to any one of claims 41 to 43, wherein said saccharide is present in said composition at 0.001-0.250 wt. %, preferably 0.001-0.200 wt. %, more preferably 0.001- 0.100 wt. %, even more preferably 0.001-0.020 wt. %, even more preferably 0.001-0.010 wt. %, even more preferably 0.002-0.010 wt. %, even more preferably 0.003-0.010 wt. %, most preferably 0.003- 0.0075 wt. %.

48. A composition for use according to any one of embodiments 41 to 47, wherein said method comprises administering an effective amount of said saccharide to said subject, preferably wherein said saccharide is administered at a daily dose as defined in any one of embodiments 27 to 34.

49. A composition for use according to any one of embodiments 41 to 48, wherein said composition is administered to said subject as defined in any one of embodiments 35 to 40.

50. A composition for use according to any one of embodiments 41 to 49, wherein said saccharide is present at a higher amount than any mammalian milk oligosaccharide present in said composition.

51. A composition for use according to any one of embodiments 41 to 50, wherein said composition: does not comprise lacto-N-neotetraose and/or lacto-N-tetraose and/or sialyllactose, preferably does not comprise lacto-N-neotetraose, more preferably does not comprise lacto-N-neotretraose and lacto- N-tetraose, even more preferably does not comprise lacto-N-neotretraose, lacto-N-tetraose and sialyllactose; or comprises any one or more of lacto-N-neotetraose, lacto-N-tetraose and sialyllactose, but at an amount in the composition which is at least 5, preferably 10, more preferably 15, even more preferably 20, even more preferably 25, even more preferably 50, times lower than the amount of said saccharide in the composition. A composition for use according to any one of embodiments 41 to 51, wherein said composition: does not comprise LNFP I and/or LNFP V and/or LNnFP V and/or LNDFH II and/or LNnDFH, preferably does not comprise LNFP I, more preferably does not comprise LNFP I and LNFP V, even more preferably does not comprise LNFP I, LNFP V and LNnFP V, even more preferably does not comprise LNFP I, LNFP V, LNnFP V and LNDFH II, most preferably does not comprise LNFP I, LNFP V, LNnFP V, LNDFH II and LNnDFH; or comprises any one or more of LNFP I, LNFP V, LNnFP V, LNDFH II and LNnDFH, but at an amount in the composition which is at least 5, preferably 10, more preferably 15, even more preferably 20, even more preferably 25, even more preferably 50, times lower than the amount of said saccharide in the composition. A composition for use according to any one of embodiments 41 to 52, wherein said composition is a pharmaceutical composition, optionally further comprising a pharmaceutically acceptable carrier, filler, preservative, solubilizer, diluent, excipient, salt, adjuvant and/or solvent. A composition for use according to any one of embodiments 41 to 52, wherein said composition is a nutritional composition, optionally further comprising a feed ingredient and/or a food ingredient, wherein said feed/food ingredient is preferably chosen from the list consisting of: a lipid, preferably one or more selected from the list consisting of an oil, fat, ester, monoglyceride, diglyceride, triglyceride and free fatty acid; a vitamin, preferably one or more selected from the list consisting of vitamin A, vitamin B, vitamin C, vitamin D, vitamin E and vitamin H, or a derivate thereof; an amino acid compound; a trace element; a mineral; an antioxidant; a prebiotic agent; a carbohydrate; an antimicrobial agent; and/or a protein. A composition for use according to embodiment 54, wherein said nutritional composition is a synthetic nutritional composition.

56. A composition for use according to embodiment 54 or 55, wherein said nutritional composition is a food product, preferably wherein said food product is selected from a list consisting of dairy product, bar, liquid product, savory snack, savory biscuit, bakery product, pasta and food supplement, more preferably wherein said food product is selected from a list consisting of dairy product, liquid product and food supplement.

57. A composition for use according to embodiment 54 or 55, wherein said nutritional composition is selected from a list consisting of infant formula, baby food, infant cereal composition, growing-up milk, milk replacer, creep feed, pet food, dry feed, frozen feed, pre weaning feed, weaning feed and post weaning feed. 8. A composition for use according to embodiment 54 or 55, wherein said nutritional composition is selected from a list consisting of dry feed, frozen feed, milk replacer, creep feed, pet food, prestarter diet, pre weaning feed, weaning feed and post weaning feed, preferably selected from a list consisting of dry feed, frozen feed, milk replacer, creep feed and pet food, more preferably wherein said nutritional composition is dry feed, frozen feed, milk replacer or creep feed, even more preferably wherein said nutritional composition is dry feed or frozen feed, most preferably wherein said nutritional composition is dry feed.

59. A method for preventing and/or treating a bacterial infection in a subject, wherein said method comprises administering a saccharide comprising a fucose, preferably a saccharide as defined in any one of embodiments 4 to 15, or a composition according to any one of embodiments 41 to 58.

60. A method according to embodiment 59, wherein said bacterium is gram-positive, preferably said bacterium is as defined in embodiment 2.

61. A method according to embodiment 59 or 60, wherein said method is for preventing and/or treating streptococcosis.

62. A method according to any one of embodiments 59 to 61, wherein said subject is as defined in any one of embodiments 16 to 25.

63. A method according to any one of embodiments 59 to 62, wherein said method comprises administering an effective amount of said saccharide to said subject, preferably wherein said saccharide is administered as defined in any one of embodiments 27 to 40.

64. Use of a saccharide comprising a fucose, preferably a saccharide as defined in any one of embodiments 4 to 15, or a composition according to any one of embodiments 41 to 58, for the manufacture of a medicament for preventing and/or treating a bacterial infection in a subject.

65. Use according to embodiment 64, wherein said bacterium is gram-positive, preferably said bacterium is as defined in embodiment 2.

66. Use according to embodiment 64 or 65, wherein said medicament is for preventing and/or treating streptococcosis. 7. Use according to any one of embodiments 64 to 66, wherein said subject is as defined in any one of embodiments 16 to 25. 8. Use according to any one of embodiments 64 to 67, wherein an effective amount of said saccharide is administered to said subject, preferably wherein said saccharide is administered as defined in any one of embodiments 27 to 40.

Definitions

The words used in this specification to describe the invention and its various embodiments are to be understood not only in the sense of their commonly defined meanings, but to include by special definition in this specification structure, material or acts beyond the scope of the commonly defined meanings. Thus, if an element can be understood in the context of this specification as including more than one meaning, then its use in a claim must be understood as being generic to all possible meanings supported by the specification and by the word itself.

The various aspects and embodiments of the invention disclosed herein are to be understood not only in the order and context specifically described in this specification, but to include any order and any combination thereof. Each embodiment as identified herein may be combined together unless otherwise indicated. All patent and literature references cited in the present specification are hereby incorporated by reference in their entirety. Unless specifically stated otherwise, all words used in the singular number shall be deemed to include the plural and vice versa. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry and nucleic acid chemistry and hybridization described herein are those well-known and commonly employed in the art. Standard techniques are used for nucleic acid and peptide synthesis. Generally, enzymatic reactions and purification steps are performed according to the manufacturer's specifications.

In the drawings and specification, there have been disclosed embodiments of the invention, and although specific terms are employed, the terms are used in a descriptive sense only and not for purposes of limitation, the scope of the invention being set forth in the following claims. It must be understood that the illustrated embodiments have been set forth only for the purposes of example and that it should not be taken as limiting the invention. It will be apparent to those skilled in the art that alterations, other embodiments, improvements, details and uses can be made consistent with the letter and spirit of the invention herein and within the scope of this invention, which is limited only by the claims, construed in accordance with the patent law, including the doctrine of equivalents. In the claims which follow, reference characters used to designate claim steps are provided for convenience of description only, and are not intended to imply any particular order for performing the steps (unless specifically stated otherwise). In this document and in its claims, the verbs "to comprise", "to have" and "to contain", and their conjugations are used in their non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. The verb "to consist essentially of" means that e.g. a composition as defined herein may comprise additional component(s) than the ones specifically identified, said additional component(s) not altering the unique characteristic of the invention. Throughout the document and claims, unless specifically stated otherwise, the verbs "to comprise", "to have" and "to contain", and their conjugations, may be preferably replaced by "to consist" (and its conjugations) or "to consist essentially of" (and its conjugations) and vice versa. In addition, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that more than one of the element is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article "a" or "an" thus usually means "at least one". The word "about" or "approximately" or "around" when used in association with a numerical value (e.g. about 10) preferably means that the value may be the given value (of 10) more or less 10%, preferably 5%, more preferably 1% of the value.

The term "polysaccharide" as used in the context of the present invention refers to a saccharide containing a plurality of repeating units comprised of simple sugars. In the context of the invention, said polysaccharide preferably has a degree of polymerization which is at least 40 (and preferably < 3000).

The terms "LNT II", "LNT-II", "LN3", "lacto-N-triose II", "lacto-N-triose II", "lacto-N-triose", "lacto-N-triose" and "GlcNAcpi-3Gaipi-4Glc" are used interchangeably.

The terms "LNT", "lacto-N-tetraose", "lacto-/V-tetraose" and "Gaipi-3GlcNAcpi-3Gaipi-4Glc" are used interchangeably.

The terms "LNnT", "lacto-N-neotetraose", "lacto-/V-neotetraose", "neo-LNT" and "Gaipi-4GlcNAcpi- 3Gaipi-4Glc" are used interchangeably.

The terms "2' fucosyllactose", "2'-fucosyllactose", "alpha-1, 2-fucosyllactose", "alpha 1,2 fucosyllactose", "a-l,2-fucosyllactose", "a 1,2 fucosyllactose", "Gaip-4(Fucal-2)Glc", 2FL" and "2'FL" are used interchangeably.

The terms "3-fucosyllactose", "alpha-1, 3-fucosyllactose", "alpha 1,3 fucosyllactose", "a-1,3- fucosyllactose", "a 1,3 fucosyllactose", "Gaip-4(Fucal-3)Glc", 3FL" and "3-FL" are used interchangeably. The terms "difucosyllactose", "di-fucosyllactose", "lactodifucotetraose", "2',3-difucosyllactose", "2', 3 difucosyllactose", "a-2', 3-fucosyllactose", "a 2', 3 fucosyllactose, "Fucal-2Gaipi-4(Fucal-3)Glc", "DFLac", 2', 3 diFL", "DFL", "DiFL" and "diFL" are used interchangeably.

The terms "2'-fucosyl-N-acetyllactosamine", "2'FlacNAc" oand "Fucal-2Gaipi-4GlcNAc" are used interchangeably.

The terms "3-fucosyl-N-acetyllactosamine", "3FlacNAc" and "Gaipi-4(Fucal-3)GlcNAc" are used interchangeably. The terms "difucosyl-N-acetyllactosamine", "diFlacNAc" and "Fucal-2Gaipi-4[Fucal-3]GlcNAc" are used interchangeably.

The terms "2'-fucosyllacto-N-biose", "2'FLNB" and "Fucal-2Gaipi-3GlcNAc" are used interchangeably.

The terms "4-fucosyllacto-N-biose", "4FLNB" and "Fucal-4[Gaipi-3]GlcNAc" are used interchangeably.

The terms "difucosyllacto-N-biose", "diFLNB" and "Fucal-4[Fucal-2Gaipi-3]GlcNAc" are used interchangeably.

The terms "LNFP-I", "lacto-N-fucopentaose I", "LNFP I", "LNF I OH type I determinant", "LNF I", "LNF1", "LNF 1" , "Blood group H antigen pentaose type 1" and "Fuc-al,2-Gal-pi,3-GlcNAc-pi,3-Gal-pi,4-Glc" are used interchangeably.

The terms "Gal-LNFP-I", "blood group B antigen hexaose type I" and "Gal-al,3-(Fuc-al,2)-Gal-pi,3- GlcNAc-pi,3-Gal-pi,4-Glc" are used interchangeably.

The terms "GalNAc-LNFP-l", "blood group A antigen hexaose type I", and "GalNAc-al,3-(Fuc-al,2)-Gal- pi,3-GlcNAc- pi,3-Gal-pi,4-Glc" are used interchangeably.

The terms "LNFP-II", "lacto-N-fucopentaose II" and "Gal-pi,3-(Fuc-al,4)-GlcNAc-pi,3-Gal-pi,4-Glc" are used interchangeably.

The terms "LNFP-III", "lacto-N-fucopentaose III" and "Gal-pi,4-(Fuc-al,3)-GlcNAc-pi,3-Gal-pi,4-Glc" are used interchangeably.

The terms "LNFP-V", "lacto-N-fucopentaose V" and "Gal-pi,3-GlcNAc-pi,3-Gal-pi,4-(Fuc-al,3)-Glc" are used interchangeably.

The terms "LNDFH I", "Lacto-N-difucohexaose I", "LNDFH-I", "LDFH I", "Le ^b -lactose", "Lewis-b hexasaccharide" and "Fuc-al,2-Gal-pi,3-[Fuc-al,4]-GlcNAc-pi,3-Gal-pi,4-Glc" are used interchangeably. The terms "LNDFH II", "Lacto-N-difucohexaose II", "Lewis a-Lewis x", "LDFH II" and "Fuc-al,4-(Gal-pi,3)- GlcNAc-pi,3-Gal-pi,4-(Fuc-al,3)-Glc" are used interchangeably.

The terms "lewis b-lewis x" and "Fucal,4-[Fuc-al,2-Gaipi,3]-GlcNAc-pi,3-Gal-pi,4-[Fuc-al,3]- Glc are used interchangeably.

The terms "MFLNH III", "monofucosyllacto-N-hexaose-lll" and "Gal-pi,4-[Fuc-al,3]-GlcNAc-pi,6-[Gal- pi,3-GlcNAc-pi,3]-Gal-pi,4-Glc" are used interchangeably.

The terms "DFLNH (a)", "difucosyllacto-N-hexaose (a)" and "Gal-pi,4-[Fuc-al,3]-GlcNAc-pi,6-[Fuc-al,2- Gal-pi,3-GlcNAc-pi,3]-Gal-pi,4-Glc" are used interchangeably.

The terms "DFLNH", "difucosyllacto-N-hexaose" and "Gal-pi,4-[Fuc-al,3]-GlcNAc-pi,6-[Fuc-al,4-[Gal- pi,3]-GlcNAc-pi,3]-Gal-pi,4-Glc" are used interchangeably.

The terms "TFLNH", "trifucosyllacto-N-hexaose" and "Gal-pi,4-[Fuc-al,3]-GlcNAc-pi,6-[Fuc-al,4-[Fuc- al,2-Gal-pi,3]-GlcNAc-pi,3]-Gal-pi,4-Glc" are used interchangeably.

The terms "LNnFP I", "Lacto-N-neofucopentaose I" and "Fuc-al,2-Gal-pi,4-GlcNAc-pi,3-Gal-pi,4-Glc" are used interchangeably.

The terms "LNFP-VI", "LNnFP V", "lacto-N-neofucopentaose V" and "Gal-pi,4-GlcNAc-pi,3-Gal-pi,4-(Fuc- al,3)-Glc" are used interchangeably.

The terms "LNnDFH", "Lacto-N-neoDiFucohexaose", "Lewis x hexaose" "Gal-pi,4-(Fuc-al,3)-GlcNAc-pi,3- Gal-pi,4-(Fuc-al,3)-Glc" are used interchangeably.

The term "cultivation" refers to the culture medium wherein the cell is cultivated or fermented, the cell itself, and the saccharide(s) that is/are produced by the cell in whole broth, i.e. inside (intracellularly) as well as outside (extracellularly) of the cell.

Brief description of the drawings

The following drawing will serve as further illustration and clarification of the present invention and is not intended to be limiting in any way.

Figure 1. Percentage of alive fish (Tilapia) infected with S. agalactiae lb at the end of the 21-day challenge period. Group A (placebo) counted 13 alive fish (n=13), group B (3-FL treatment) counted 13 alive fish (n=13), group C (2'FL treatment) counted 10 alive fish (n=10). "*" refers to a statistically significant difference between the 3-FL treatment group and placebo group of P < 0.05 (Chi-squared test).

Examples

The invention will be described in more detail in the examples. The following examples will serve as further illustration and clarification of the present invention and are not intended to be limiting in any way.

Example 1: 3-FL and 2-FL

3-fucosyllactose (3-FL) was recombinantly produced in E. coli as described in example 16 of 2020/127417 (Helicobacter pylori alpha-1, 3-fucosyltransferase), purified as described in examples 13-20 of WO 2022/034079, and subsequently dried to obtain 3-FL powder (purity 96.75 %) as described in WO 2019/160922 (Example 7).

2'-fucosyllactose (2'FL) was recombinantly produced in E. coli and purified as described in examples 10, 13 and 14 of WO 2022/034079, and subsequently dried as described in Example 21 of WO 2022/034079 to obtain 2'FL powder (purity 89.6 %).

Example 2: Streptococcus aqalactiae challenge in fish (Tilapia)

Experimental design

The experiment was carried out at the facility of Ictyopharma fish CRO (Deneuille-Les-Mines, France) involving 15 tanks of 100L. Independent recirculation systems were used, each containing 5 tanks. Each system was equipped with a mechanical filter and a bio-filter as well as degassing columns and UV. The following water parameters were measured on a daily basis: dissolved oxygen (DO), pH and temperature. NH ₄ and NO2 were measured at least 2 times a week. Water salinity is at 0 ppt, temperature 28°C ± 1,5°C, exchange rate 200L/h. A summary of the water quality parameters during the trial is provided in Table 1.

Table 1 - Water quality

Tilapia (Oreochromis niloticus), majority YY, weighted at start of trial around 17.66±0.26 g are experimental species. A total of 435 fish were used in groups of 29 animals. Each group (n = 29) was housed in a tank with a unique identification. Fish were not marked individually. The tank was used as the experimental unit. In each tank a total of 29 fish were placed at the beginning of the acclimatization period. The animals were acclimatized to their tanks 4 days before the start of the experiment. During this phase, all animals were fed the control feed (see Table 2). In the experiment (4 weeks of feeding followed by a Streptococcus agalactiae challenge for 3 weeks), one third of the fish were treated with 2'FL (i.e. 5 tanks; group C), i.e. receiving the 2'FL feed (Table 2), one-third of the fish were treated with 3-FL (i.e. 5 tanks; group B), i.e. receiving the 3-FL feed (Table 2), whereas the last one-third (i.e. 5 tanks; group A) served as control, i.e. receiving the control feed. Animals were fed ad libitum according to their mean body weight but using a maximum feeding rate: week 1 (5% body weight), week 2, 3 and 4 (4.5% body weight). Feeding rates are adjusted weekly based on mean individual weight per tank. After experimental infection with Streptococcus agalactiae (see further), fish are fed ad libitum. The supplementation dose of 3-FL (or 2'FL) for week 1 and for week 2-4 are 25 mg and 22.5 mg per kg body weight per day, respectively. Our data showed that each tilapia (starting weight as aforementioned) eats on average 1.3 g/day (i.e. 0.65 mg 3-FL or 0.65 mg 2'FL) within the first 4 weeks.

At the end of the 4 weeks feeding, one day prior to challenge, three fish per tank (of 29 fish) were removed. One "spare" fish per tank was removed the morning of challenge. If more than one mortality occurred in a tank during the feeding stage, "spare" fish from other tanks from the same treatment were introduced the morning of challenge to ensure 25 fish per tank at the start of the challenge period.

One glycerol stock (S. agalactiae serotype lb, Ictyopharma strain TI063) was recovered from the freezer <-50 ° C. The tube was thawed at room temperature and 1 ml inoculated in IL of tryptone soy broth (TSB) and incubated with agitation by magnetic bar at 30°C overnight. After incubation for approximately 15h- 18h, with OD660nm > 0.8 (corresponding to approx. 10 ⁹ CFU/ml) the suspension was used for immersion infection. Challenge was conducted using 30ml of overnight culture per tank. CFU of this culture was verified on tryptic soya agar (TSA), as expected, to be 2.8xl0 ⁹ CFU/ml. Experimental infection was performed by immersion. The fish remained in static conditions with air supply for 1 hour after which the water recirculation was turned back on. All systems had a UV light that decreases the pathogen load sufficiently not to interfere with the fish to fish contaminations in the individual tanks. The infection progressed from fish to fish in a natural way, closest to natural infection occurring in the field.

Diets

Experimental diets were manufactured by SPAROS Lda. (Olhao, Portugal). Powder ingredients were grinded (below 200 micron) in a micropulverizer hammer mill (Hosokawa Micron, SHI, The Netherlands). Diets were manufactured by temperature extrusion (pellet size: 1.5 mm) by means of a low shear extruder (Italplast P55, Italy). Upon extrusion, all feed batches were dried. Extrusion temperatures used are generally in between 105-115 ⁹ C and drying temperature around 120°C for approximately 1.5 min (initial section of the dryer) and then a gradual decrease towards the end of the dryer, at which temperature range 40-45 ⁹ C. The basal diet was formulated to meet or exceed the nutrient requirement for Tilapia. The diet composition and calculated nutrient contents are shown in table 2. 3-FL (or 2'FL) was diluted in a small amount of water and subsequently emulsified in oil (0.05 % w/v of 3-FL or 0.05 % w/v of 2'FL) before top- dressed onto the post-extruded feed under vacuum (i.e. forming the top-coating). The top-coating of the diet for the placebo group did not contain 3-FL or 2'FL. Diets were packed in plastic sealed bins and transferred to the experimental site.

Table 2 - diet composition

* 0.05 % (w/w) means that 0.5 g 3-FL is present per kg feed

** 0.05 % (w/w) means that 0.5 g 2'FL is present per kg feed

Quantification of Streptococcus aaalactiae load in the brain

At the conclusion of the 21-day challenge period, one entire tank per treatment (i.e. group A, group B and group C) was sampled to quantify the pathogen load of the surviving fish. Tanks were chosen at random. Entire brains were collected from euthanized fish using aseptic technique to minimize the possibility of contamination. Brains were immediately weighed in sterile microcentrifuge tubesand diluted with 9x volume of sterile saline solution. Brains from visually moribund fish were identified with an identifying mark/sticker on the microcentrifuge tube. Once diluted, brains were mechanically emulsified as much as possible with the use of a sterile microcentrifuge pestle. After mechanical breakdown of tissue, samples were centrifuged at 2000 rpm for 15 seconds. Serial dilutions using the supernatant were conducted taking 100 pl and adding to 900 pl of sterile saline solution.

Spread plates of the 4 most relevant dilutions (10 ¹ - 10' ⁴ for surviving fish, 10' ³ -10 ^-6 for visually moribund) were plated in duplicate and incubated for 48 hours at 30°C on CNA media (specific for gram positive bacteria). Plate counts were conducted to calculate CFU/g of brain tissue. Colonies of Streptococcus agalactiae serotype lb were visually identified by experienced lab technicians. Colonies (5% per plate) counted were tested via agglutination for confirmation of the identity of the pathogen using the GBS latex agglutination test (SSI Diagnostica, Denmark).

Results

Generally, S. agalactiae will spread around the fish body upon infection. Mortality usually peaks within 2- 3 weeks which was also the case in present experiment. Mortality was recorded daily per tank and per condition after challenge. Up to end point, the control treatment showed accumulated mortality of 54.4%±5.4 while the 3FL-treatment and the 2'FL treatment achieved accumulated mortality of 57.6%±11.5 and 56.0%±10.2, respectively. Analysis of survival kinetics using Kaplan-Meier (a nonparametric statistic used to estimate the survival function from survival data) showed no significant difference between the control group and the 3FL or 2'FL treatments at the 95% or 90% confidence interval (P-value = 0.3559). Figure 1 shows the percentage of fish in one tank that retains Streptococcus agalactiae after 3 weeks upon challenge with S. agalactiae. Out of 13 alive fish in the control group (group A), six were found with S. agalactiae in the brain (46.15%). The pathogens were found in the brains of many fish in tanks treated with 2'FL (group C) at the ratio of 90%. In contrast, significantly less fish in the tank treated with 3-FL (group B) retain S. agalactiae in the brain (30.77%) as compared to the control group (Chi-squared test; P-value = 0.021). This experiment hence clearly shows that 3-FL is an efficacious agent to prevent and/or treat streptococcosis in fish.

List of references

Derya et al (2020), Biotechnologically produced fucosylated oligosaccharides inhibit the binding of human noroviruses to their natural receptors, J. Biotech. 319: p. 31-38.

Dumon et al (2001), In vivo fucosylatoin of lacto-N-neotetraose and lacto-N-neohexaose by heterologous expression of Helicobacter pylori alpha-1,3 fucoysltransferase in engineered Escherichia coli, Glyconj. J. 18(6): p. 465-474.

Huang et al (2021), Sulfo-fluorous tagging strategy for site-selective enzymatic glycosylation of para-human milk oligosaccharides, ACS Catal. 11(5): p. 2631-2643.

Lu et al (2021), Engineered microbial routes for human milk oligosaccharide synthesis, ACS Synth. Biol. 10(5): p. 923-938.

Mishra et al (2018), Current challenges of Streptococcus infection and effective molecular, cellular and environmental control methods in aquaculture, Mol. Cells 41(6): p. 495-505.

Morales-Covarrubias et al (2018), Streptococcus penaeicida sp. nov., isolated from a diseased farmed pacific white shrimp (Penaeus vannamei), Int. J. Syst. Evol. Microbiol. 68(5): p. 1490-1495.

- W02010/037785

- WO2019/160922

- W02020/115671

- WO2021/067641

- WO2021/242866

- W02022/034073

- WO 2022/034075

- WO 2022/034077

Yu et al (2017), Helicobacter pylori alphal-3/4-fucosyltransferase (Hp3/4FT)-catalyzed one-pot multienzyme (OPME) synthesis of lewis antigens and human milk fucosides, Chem Comm. 53(80): p. 11012-11015.

Zhou et al (2021), Biotechnological production of 2'-fucosyllactose : a prevalent fucosylated human milk oligosaccharide, ACS Synth. Biol. 10(3): p. 447-458.