LU101096B1 - Natural non-pathogenic microorganisms capable of associating glycolipids or lipopeptides and use thereof - Google Patents

Abstract

The present invention relates to natural non-pathogenic microorganisms capable of associating glycolipids and/or lipopeptides such as e.g. binding peptides (e.g. linear or circular binding peptides, bi-cyclic peptides including a stabilizing molecule, DARPins), immunological active peptides, proteins (e.g. anti-inflammatory cytokines, binding molecules), antibodies, antibody fragments (e.g. single domain antibodies, single chain antibdoies scFv) or derivatives thereof coupled to a lipid carrier and uses thereof. The present invention further relates to naturally-occurring non-pathogenic microorganisms (e.g., bacteria, archae, yeasts or fungi) capable to physically interact with a lipid carrier (e.g., glycolipid, lipopeptide etc.) and present the carried moiety (e.g., carbohydrate, peptide or chemical molecule, etc.) to the environment (e.g., exterior surface of the cell). Among others, the invention is useful for the development of oral vaccines, oral drug delivery systems and anti-infectious agents (e.g., high-density natural display system) as well as for various topical applications and/or to inhale and/or compositions (e.g., medical and/or pharmaceutical applications and/compositions) including such for topical or inhalation applications and/or compositions for use in treatment or prevention of diseases such as diseases from the gut, digestive tract, oral cavity, skin, urogenital tract, lung and/or other sites. Furthermore, microorganisms, compositions, vaccines, adjuvants, kits, methods and uses of the present invention are useful for immunization, delivery, screening and targeting applications, e.g., particularly relating to pharmaceutically active compounds, toxins and pathogenic microorganisms. Furthermore, the present invention relates to methods of associating and/or loading and methods of displaying carbohydrates or lipopetides on microorganisms.

Description

New LU-patent application | _ LU101096 Applicants: ULSEMER Philippe; TOUTOUNIAN Kawe, GOLETZ Steffen, GÔTZ Peter Our ref.: AMM16410LU Natural non-pathogenic microorganisms capable of associating glycolipids or lipopeptides and use thereof

FIELD OF THE INVENTION

[001] The present invention relates to natural non-pathogenic microorganisms capable of associating glycolipids and/or lipopeptides, such as e.g. comprising binding peptides (e.g. linear or circular binding peptides, bi-cyclic peptides including a stabilizing molecule, DARPins), immunological active peptides, proteins (e.g. anti-inflammatory cytokines, binding molecules), antibodies, antibody fragments (e.g. single domain antibodies, single chain antibdoies scFv) or derivatives thereof coupled to a lipid carrier, and uses thereof. The present invention further relates to naturally-occurring non-pathogenic microorganisms (e.g. bacteria, archae, yeasts or fungi) capable to physically interact with a lipid carrier (e.g. glycolipid, lipopeptide etc.) and present the carried moiety (e.g., carbohydrate, peptide or chemical molecule, etc.) to the environment (e.g., exterior surface of the cell). Among others, the invention is useful for the development of oral vaccines, oral drug delivery systems and anti-infectious agents (e.g., high-density natural display system) as well as for various topical applications and/or to inhale and/or compositions (e.g., medical and/or pharmaceutical applications and/compositions) including such for topical or inhalation applications and/or compositions for use in treatment or prevention of diseases such as diseases of the gut, digestive tract, oral cavity, skin, urogenital tract, lung and/or other sites. Furthermore, microorganisms, compositions, vaccines, adjuvants, kits, methods and uses of the present invention are useful for immunization, delivery, screening and targeting applications, e.g., particularly relating to pharmaceutically active compounds, toxins and pathogenic microorganisms. Furthermore, the present invention relates to methods of associating and/or loading and methods of displaying carbohydrates or lipopetides on microorganisms.

BACKGROUND OF THE INVENTION

[002] Non-pathogenic microorganisms especially food graded microorganisms displaying a specific structure (e.g., oligosaccharide, peptide, proteins) on their surface have a broad application potential in food and medicine like oral vaccines, oral drug delivery systems and anti-infectious agents. Same also holds true for bacteria of the skin and other sites such urogenital tract, respiratory system or oral cavity. 1

[003] Standard procedures to produce non-pathogenic microorganisms displaying a LU101096 specific structure (e.g., oligosaccharide or peptide) on their surface are mainly based on genetic modifications that result in recombinant microorganisms (Genetically Modified Organisms or GMOs). Alternatively, non-GMO approaches are using fusion proteins | 5 composed of a peptide of interest bound to a protein-domain (anchor-domain) known to anchor itself to the surface of suitable microorganisms (Michon et al., 2016). However, when loaded on live microorganisms, the stability and integrity of the fusion protein may be affected by natural proteolytic activities of the said microorganism (Ganesh et al., 2014). Furthermore, potential immunogenic properties of the anchor-domain may significantly limit the application potential of this methodology (Schmidt et al., 2011).

[004] Thus, as is evident from the above, to obtain a non-pathogenic microorganism displaying a non-endogenous structure (e.g., peptide or carbohydrate) to the environment, the prior art provides either recombinant microorganisms, the use of which is related to significant safety concerns, or recombinant fusion proteins that may be loaded on non- pathogenic microorganisms. Currently, no non-pathogenic microorganisms are known to be able to bind a heterologous lipid carrier (e.g., glycolipid or lipopeptide) in such a way so that the carried moiety (e.g., carbohydrate or peptide) is displayed to the environment.

[005] However, such a strategy would be particularly promising to generate non-pathogenic microorganisms presenting oligosaccharides, peptides or chemical molecules of interest to the environment without employing genetic modifications of the non-pathogenic mircroorganism. The application’s potential includes, among others, the development of oral vaccines, oral drug delivery systems, anti-infectious agents and functional foods.

SUMMARY OF THE INVENTION

[006] The present invention further relates to modified microorganisms comprising: a cell and a heterologous lipid carrier comprising: (a) a lipid portion, wherein said lipid portion is at least partially associated with an exterior surface of said cell of said modified microorganism, wherein said lipid portion comprising: a ceramide-like glycolipid moiety and/or a fatty acid moiety; and (b) a non-lipid portion, wherein said microorganism is capable of locating and/or displaying said non-lipid portion or fragment thereof onto the exterior surface of said cell; uses and methods of productions thereof.

[007] As explained above there is a need for new therapeutic interventions to prevent and/or treat toxin-related diseases, conditions, states or syndromes particularly associated with bacterial infections, viral, fungal or autoimmune diseases.

[008] The present application satisfies this demand by the provision of modified microorganisms, compositions, vaccines, adjuvants, kits, methods and uses described herein below, characterized in the claims and illustrated by the appended Examples and Figures.

2

BRIEF DESCRIPTION OF THE DRAWINGS LU101096

[009] Figure 1: Binding of HRP-Cholera toxin to isolates LOM, BLP, G29 and Lac9. Isolates were incubated overnight in PBS at 30°C with or without GM1 at 5ug/ml. The isolates were extensively washed. The presence of GM1 on the surface of the isolates was analyzed by ELISA using HRP-cholera toxin. As used herein, “LOM” = Lactobacillus reuteri, “BLP” = Lactobacillus paracasei, “G29” = Enterococcus faecalis, “Lac9” = Lactobacillus reuteri.

[0010] Figure 2: Stability of the association of GM1 with BLP isolates.

(A) Binding of labeled cholera toxin to the GM1-loaded strain BLP after pasteurization (15 min 70°C). (B) Binding of labeled cholera toxin to the GM1-loaded strain BLP after incubation at pH=1 for 1 hour at 37°C. (C) Binding of labeled cholera toxin to the GM1-loaded strain BLP after freezing and thawing. (D) Binding of labeled cholera toxin to the GM1-loaded strain BLP after freezing drying and re-hydration. (E) Binding of labeled cholera toxin to the GM1- loaded strain BLP after 4 hours incubation at 37°C. (F) Binding of labeled cholera toxin to the GM1-loaded strain BLP after incubation in gastric juice for 30 min at 37°C.

DETAILED DESCRIPTION OF THE INVENTION

[0011] The following detailed description refers to the accompanying Examples and Figures that show, by way of illustration, specific details and embodiments, in which the invention may be practiced. These embodiments are described in sufficient detail to enable those skilled in the art to practice the invention. Other embodiments may be utilized such that structural, logical, and eclectic changes may be made without departing from the scope of the invention. Various aspects of the present invention described herein are not necessarily mutually exclusive, as aspects of the present invention can be combined with one or more other aspects to form new embodiments of the present invention.

[0012] Definitions:

[0013] Unless otherwise specified, the terms used herein have their common general meaning as known in the art (e.g., as defined by IUPAC. Compendium of Chemical Terminology, 2nd ed. (the "Gold Book", compiled by A. D. McNaught and A. Wilkinson. Blackwell Scientific Publications, Oxford (1997); XML on-line corrected version: http://goldbook.iupac.org (2006-) created by M. Nic, J. Jirat, B. Kosata; updates compiled by A. Jenkins. ISBN 0-9678550-9-8. https://doi.org/10.1351/goldbook).

[0014] As used herein, the term “microorganism” may refer to any organism too small to be viewed by the unaided eye, e.g., bacteria, virus, protozoa, archae, fungi and algae. Unless defined otherwise, the term "microorganism(s)" used herein includes bacteria, viruses (e.g., 3

Norovirus, Herpes simplex virus 1 and 2 (HSV-1 and HSV-2), influenza virus, respiratory LU101096 syncytial virus Human papilloma virus (HPV) etc.), fungi (including unicellular and filamentous fungi), yeasts, protozoa and multi-cellular parasites. Typical sources of microorganisms described herein include faeces, gut, skin, nose, ear, mouth, eye, urogenital or respiratory tract, breast milk, foods (including but not limited to: milk products, meet, etc.), pure cultures, soil, water and plants.

[0015] Exemplary non-limiting microorganisms include bacteria (e.g., gram-positive or gram- negative (e.g., from genus Bacteroides, e.g., B. vulgatus, B.xylanisolvens, B. fragilis) bacteria); gram-positive bacteria include bacteria selected from the genera consisting of: Lactobacillus (e.g., Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus salivarius, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus or Lactobacillus vaginalis), Bifidobacterium (e.g., Bifidobacterium breves Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium infantis or Bifidobacterium pseudocatenulatum), Clostridium (e.g. Clostridium perfringens, Clostridium coccides), Enterococcus (e.g., Enterococcus avium, Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinarum, Enterococcus casseliflavus or Enterococcus sp.), Pediococcus (e.g., Pediococcus pentosaceus) and Streptococcus (e.g., Streptococcus salivarius or Streptococcus vestibularis); fungi including fungi selected from the group consisting of: Candida yeasts, Saccharomyces yeasts (e.g., S. cerevisiae and S. pombe) and yeasts in the family Dipodascaceae; further preferably said Dipodascaceae yeasts are Galactomyces, Geotrichum or Saprochaete yeasts; and protozoa (e.g., Chilomastix mesnili, Endolimax nana, Entamoeba coli, Entamoeba dispar, Entamoeba hartmanni, Entamoeba polecki or lodamoeba buetschlii). Further exemplary non-limiting gram-negative bacteria include bacteria of Enterobacteriaceae family, e.g., genera Escherichia (e.g., Escherichia sp., E. coli, E. fergusonii), Klebsiella (e.g., K. pneumoniae, Klebsiella sp.), Enterobacter (e.g., E. cloacae, Enterobacter sp.), Proteus (e.g., P. mirabilis, Proteus sp.), Morganella (e.g., M. morganii Morganella sp.), Citrobacter (e.g., Citrobacter sp.), Serratia (e.g., Serratia sp.).

[0016] As used herein, the term “pathogenic microorganism” may refer to microorganisms, which have (i.e., express and secrete) a toxin, include microorganisms as described herein that can cause lesion and/or disease of mucosa (e.g., virus, e.g., Norovirus, Rotavirus, Herpes simplex virus 1 and 2 (HSV-1 and HSV-2), Influenza virus, Respiratory syncytial virus, Human papilloma virus (HPV) etc., protozoa (e.g., Giardia, Cryptosporidium spp. and Blastocystis spp.)), including but not limited to buccal mucosa, esophageal mucosa, gastric mucosa, intestinal mucosa, nasal mucosa, olfactory mucosa, oral mucosa, bronchial mucosa, mucosa of the respiratory tract, uterine mucosa, endometrium (mucosa of the uterus), vaginal mucosa, penile mucosa by, inter alia, a toxin. "Pathogenic microorganisms" 4further include microorganisms that cause lesions and/or disease of the gastrointestinal tract LU101096 such as diarrhea.

[0017] However, preferably the obtained or obtainable modified microorganism is non- pathogenic. Nonpathogenic microorganisms preferably include, but are not limited to, microorganisms categorized as Generally Recognized As Safe (GRAS). Non-pathogenic microorganisms further preferably include but are not limited to lactic acid bacteria or bifidobacteria. They may also include opportunistic pathogenic microorganisms. Preferably, the non-pathogenic microorganism of the present invention may be used as an active ingredient (e.g., an active agent) in a pharmaceutical composition (e.g., it can be used live or pasteurized or otherwise inactivated). It can also be used in a composition comprising only one type of non-pathogenic microorganism or more than one microorganism, e.g., and each of such a microorganism can carry one or more type of glycolipid or lipopeptide.

[0018] As used herein, the term “exterior surface” includes any localization accessible from the outside of the microorganism of the present invention. Preferably, said localization may be directly exposed to the environment, i.e., be in direct contact with the environment, or may be in indirect contact, i.e., be in indirect contact with the environment, e.g. via pores (e.g. porins) that connect the periplasm with the environment. Hence, the non-lipid portion of the lipid carrier of the present invention may be associated with (e.g., present) or integrated in the inner membrane, outer membrane, cell wall, cell membrane, polysaccharide (e.g. capsule polysaccharide), pilus, flagellum or fimbria of the modified microorganism of the present invention. Further preferably the non-lipid portion of the lipid carrier of the present invention can be bound by a soluble (or non-soluble) binding molecule, receptor or toxin (e.g., in the outer membrane, cell wall, cell membrane, polysaccharide (e.g., capsule polysaccharide), pilus, flagellum or fimbria of the modified microorganism of the present invention) or be recognized by the immune system generating or blocking a specific immune response.

[0019] The terms "isolated" or "isolation" as used herein, refer to the separation of the obtained microorganism from the starting composition comprising a plurality of microorganisms, e.g. a sample of the natural environment of the microorganism or any other used source material. A further example of an isolated microorganism of the present invention may refer to a microorganism isolated through (e.g., by the means of or as part of) any screening application/s (e.g., a method and/or use) as described herein, e.g., as a positive binder/associator (e.g., from a culture of microorganisms (e.g., “pure culture”, e.g., a population of cells or multicellular organisms growing in the absence of other species or types) source material). 5

[0020] As used herein, the term “heterologous” may refer to an entity (e.g., lipid carrier or LU101096 chemical compound or substance or molecule) of an origin different from the cell of the microorganism of the present invention (e.g., said entity is not expressed or synthetized by said microorganism).

[0021] As used herein, the term “lipid” may refer to a substance of biological and synthetic origin that is soluble in nonpolar solvents. Exemplary non-limiting lipids of the present invention include saponifiable lipids, such as, but not limited to, glycerides (e.g., fats and oils) and phospholipids, as well as non-saponifiable lipids and steroids, ceramide-like glycolipids, fatty acids, amino alcohols (e.g., sphingosines), sphingolipids.

[0022] As used herein, the term “carrier” may refer to a diluent, adjuvant, excipient, or vehicle with which a substance is administered. Exemplary carrier in the sense of the present invention is a lipid carrier comprising: a lipid portion, wherein said lipid portion is at least partially associated with an exterior surface of said cell of said modified microorganism, wherein said lipid portion comprising: a ceramide-like glycolipid moiety (e.g., comprising a ceramide moiety, a sphingolipid moiety or sphingosine moiety) and/or a fatty acid moiety; preferably said exterior surface of said cell comprising: a cell wall and/or a cell membrane and/or an outer cell membrane and/or a polysaccharide (e.g., capsule polysaccharide); further preferably said lipid portion is at least partially incorporated and/or adhered and/or bound to said exterior surface of said cell; and a non-lipid portion, wherein said microorganism is capable of locating and/or displaying said non-lipid portion or fragment thereof (e.g., a peptide (e.g., an immunologically active peptide) or a protein (e.g., an anti- inflammatory cytokine), an antibody (e.g., a single domain antibody) or a fragment thereof, and other derivatives (e.g., haptens, or parts of pathogenic organisms (for vaccination)) coupled to the lipid portion) onto the exterior surface of said cell, preferably said non-lipid portion comprising a carbohydrate moiety and/or lipopeptide moiety (e.g., a glycosylated lipopeptide moiety, e.g., a carbohydrate antigen); further preferably said non-lipid portion is capable of binding and/or reducing toxicity and/or neutralizing a toxin.

[0023] Further, exemplary non-limiting lipid carriers of the present invention may have one or more of the following characteristics: comprising a glyceride moiety; preferably said glyceride moiety comprises at least one fatty acid, further preferably said lipid portion of said lipid carrier comprises said glyceride moiety, comprising a ceramide moiety, wherein preferably: (a) said ceramide moiety is composed of an amino alcohol (e.g., sphingosine) and/or a fatty acid, further preferably said lipid portion of said lipid carrier comprises said ceramide moiety; and/or (b) said ceramide moiety is a B-anomerically linked moiety or a-anomerically linked moiety, most preferably said lipid portion of said lipid carrier comprises said ceramide moiety; comprising a carbohydrate moiety; preferably said non-lipid portion of said lipid carrier comprises said carbohydrate moiety; further preferably said carbohydrate moiety is R- (e.g., 6the first sugars linked to ceramide are typically B-linked galactose (GalCer) or glucose LU101096 (GlcCer)) or a- linked (e.g., for increasing immunity) to said ceramide moiety, further preferably said carbohydrate is not a monosaccharide or a disaccharide moiety; further preferably said carbohydrate is selected from the group consisting of: an oligosaccharide and a polysaccharide, most preferably the first sugar of the said carbohydrate is a galactose, a glucose, a mannose, a xylose, a neuraminic acid, a N-acetyl glucosamine, N-acetyl galactosamine or a galacturonic acid; comprising one or more polypeptides (e.g., a heterologous recombinant or fusion polypeptide, e.g., a glycosylated polypeptide or an immunologically active polypeptide); preferably said non-lipid portion comprising said one or more polypeptides; further preferably said one or more polypeptides is an enzyme, a cytokine or a chemokine, a peptidomimetic compound, an antigen, an antibody (e.g., a single chain or a single domain antibody), a fragment or derivative thereof: comprising a pharmaceutically active compound; preferably said non-lipid portion comprising said pharmaceutically active compound; is not comprising a recombinant and/or fusion polypeptide (e.g., an endogenous recombinant and/or fusion polypeptide), preferably said recombinant and/or fusion polypeptide is obtained by the means of artificial genetic manipulation; is not expressed or synthetized by said microorganism; is at least partially expressed or synthetized by said microorganism (e.g., endogenously synthetized), preferably said microorganism is capable of expressing or synthetizing a ceramide or sphingolipid moiety (e.g., a Sphingomonas sp. Gram-negative bacterium); said lipid portion is not comprising polypeptides; is not comprising a transmembrane polypeptide or a polypeptide membrane anchor domain; is not susceptible to proteolysis (e.g., under physiological conditions); is not immunogenic to a mammalian host, preferably said mammalian host is human; is immunogenic to a mammalian host, preferably said mammalian host is human; is not covalently bound to said cell membrane of said cell of said modified microorganism; comprising a glycolipid; comprising a lipopeptide (e.g., a glycosylated lipopeptide).

[0024] Most preferably, the lipid carrier of the present invention is selected from the group consisting of: Monosialotetrahexosylganglioside (GM1), Globotriaosylceramide (Gb3), Ganglioside GD1a, Gangliosides GM2, GD2, GD1b, GT1b, GT1c, GQ1c, GA1, GM1b, Gangliosides GM3, GD3 und GT3, Gangliosides Gb4, Blood Group Type |, Type 2, Blood Group A, Blood Group B, Blood Group H, Blood Group H Type 1, Blood Group H Type 2, Blood Group H Type 3, Lewis y, Lewis a, Lewis b, Lewis x, H, Sialyl Lewis x, Sialyl Lewis a, Sialyl Lewis b, Sialyl Lewis x, alpha Gal epitope, Gal a1-3Galf31-4GlacNAc, wherein:

[0025] GM2-1 = aNeu5Ac(2-3)bDGalp(1-?)bDGalNAc(1-?)bDGalNAc(1-?)bDGlcp(1-1)Cer

[0026] GM3 = aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer 7

[0027] GM2,GM2a(?) = N-Acetyl-D-galactose-beta-1,4-[N-Acetylneuraminidate- alpha-2,3-}- LU101096 Galactose-beta-1,4-glucose-alpha-ceramide GM2b(?) = aNeuSAc(2-8)aNeu5Ac(2- 3)bDGalp(1-4)bDGlcp(1-1)Cer

[0028] GM1,GM1a = bDGalp(1-3)bDGalNAc[aNeu5Ac(2-3)]bDGalp(1-4)bDGlcp(1-1)Cer

[0029] asialo-GM1,GA1 = bDGalp(1-3)bDGalpNAc(1-4)bDGalp(1-4)bDGlcp(1-1)Cer

[0030] asialo-GM2,GA2 = bDGalpNAc(1-4)bDGalp(1-4)bDGicp(1-1)Cer

[0031] GM1b = aNeu5Ac(2-3)bDGalp(1-3)bDGalNAc(1-4)bDGalp(1-4)bDGlcp(1-1)Cer

[0032] GD3 = aNeu5Ac(2-8)aNeu5SAc(2-3)bDGalp(1-4)bDGicp(1-1)Cer

[0033] GD2 = bDGalpNAc(1-4)[aNeuSAc(2-8)aNeu5Ac(2-3)]bDGalp(1-4)bDGlcp(1-1)Cer

[0034] GD1a = aNeuSAc(2-3)bDGalp(1-3)bDGalNAc(1-4)[aNeu5Ac(2-3)]bDGalp(1- 4)bDGlcp(1-1)Cer

[0035] GD1alpha = aNeu5Ac(2-3)bDGalp(1-3)bDGalNAc(1-4)[aNeu5Ac(2-6)]bDGalp(1- 4)bDGicp(1-1)Cer

[0036] GD1b = bDGalp(1-3)bDGalNAc(1-4)[aNeu5Ac(2-8)aNeu5Ac(2-3)]bDGalp(1- 4)bDGlicp(1-1)Cer

[0037] GT1a = aNeuSAc(2-8)aNeuSAc(2-3)bDGalp(1-3)bDGalNAc(1-4)[aNeu5Ac(2- 3)]bDGalp(1-4)bDGlcp(1-1)Cer

[0038] GT1,GT1b = aNeu5Ac(2-3)bDGalp(1-3)bDGalNAc(1-4)[aNeu5Ac(2-8)aNeu5Ac(2- 3)]bDGalp(1-4)bDGlcp(1-1)Cer

[0039] OAc-GT1b = aNeuSAc(2-3)bDGalp(1-3)bDGalNAc(1-4)aXNeu5Ac9Ac(2- 8)aNeu5Ac(2-3)]bDGalp(1-4)bDGlcp(1-1)Cer

[0040] GT1c = bDGalp(1-3)bDGalNAc(1-4)[aNeu5Ac(2-8)aNeu5Ac(2-8)aNeu5Ac(2- 3)]bDGalp(1-4)bDGlcp(1-1)Cer

[0041] GT3 = aNeu5Ac(2-8)aNeuSAc(2-8)aNeu5Ac(2-3)bDGal(1-4)bDGlc(1-1)Cer;

[0042] Gal(a 1-4)Gal(R1-4)GIcNAc-R,

[0043] Gal(a 1-4)Gal(31-4)Glc NAc-(B1-3)Gal(B1-4)GIc-R,

[0044] Gal(a 1-4)Gal(B1-4)GIcNAc(B1-2) Man-R

[0045] further, any one of the following: En — 8

Lewis x R LU101096 6'-Sulfo Sialyl 6°-Sia-6-Su- Blood Group A Blood Group A Blood Group A 9

Blood Group A LU101096

Type 2 GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-4)GIicNAc(B1-)-R

Blood Group A

Type 2 (difucosyl) | GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-4)[Fuc(a 1-3)]GIcNAc(B1-)-R

Blood Group A

Type 3 GaiNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-3)GalNAc(a 1-)-R

Blood Group A

Type 4 GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-3)GalNAc(B1-)-R

Blood Group B Gal(a 1-3)[Fuc (a 1-2)]Gal(R1-3)GIcNAc(R1-3)Gal-R,

Biood Group B

Type 2 Gal(a 1-3)[Fuc(a 1-2)]Gal(R’1-4)GIcNAc(R1-)-R

Blood Group H

Type 1, Fuc(a 1-2)Gal(R1-3)GIcNAc(B1-)-R

Blood Group H

Type 2, Fuc(a 1-2)Gal(R1-4)GIcNAc(B1-)-R

Blood Group H

Type 3, Fuc(a 1-2)Gal(R1-3)GalNAc(a 1-)-R

Blood Group H, Fuc(a 1-2)Gal(R1-)-R

Sia6'H (type 2) Neu5Aca2-6(Fuca1-2)GalB1-4GIcNAcB-R

6-LacNAc-TF GalB1-4GIcNAcB1-6(GalB1-3)GalNAca-R

C-Series Ganglio-

sides Oligo-

saccharide /

Ganglio-tetraosyl | R-Gal(R1-3)GalNAc(B1-4)[Neu5Ac(a 2-8)Neu5Ac(a 2-8)NeuSAc(a 2-

Core Structure 3)IGal(R1-4)Glc(R1-1)-R

C-Series Ganglio-

sides Oligo-

saccharide /

Hemato- or

Ganglio-Type NeubAc(a 2-8)Neu5Ac(a 2-8)Neu5Ac(a 2-3)Gal-R

Cyclic Sialyl 6- LU101096 Sulfo Lewis x cyclicNeuSAc(a 2-3)Gal(b1-4)[Fuc(a 1-3)][HSO3(-6)]GIcNAc-R Gal(B1-4)[Fuc(a 1-3)]GIcNAc(R1-3)Gal(b1-4)[Fuc(a 1-3)JGIcNAc(B1-)-R Disialyl Lewis a NeusSAc(a 2-3)Gal(B1-3)[Neu5Ac(a 2-6)][Fuc(a 1-4)]GIcNAc(B1-)-R Disialyl Lewis c NeuSAc(a 2-3)Gal(R1-3)[Neu5Ac(a 2-6)JGIcNAc(B1-)-R

F1 Alpha Gal(b1-4)GIcNAc(b1-6)GalNAc(a 1-)Ser/Thrfucosyl GM1 (Fucal-2Gal31-3GalNAcR1-4[NeuAca2-3]-Gal1-4GlcR1-I-R

GA1: (Gg4Cer) GalB1,3GalNAcB1,4GalB1,4GIcB1-R

GA2: (Gg3Cer) GalNAcB1,4GalB1,4GIcB1-R

Gal a1-3Galß1-

Gal a 1-3GalB1-4GlacNAc - R 4GlacNAc Galili, Gala- Gala1-3GalB1-4GIcNAcB-R

3'LacNAc er

GalNAc-GD1a:

(IV3Neu5AclII3Neu

5AcGg5Cer) GalNAcB1,4GalB1,3GalNAcB1, 4(Neu5Aca2,3)GalB1, 4GlcB1-R GalR1,3GalNAcB1,4GalB1,4GlcfB - R

GbOse3Cer:

(Gb3Cer) Gala1,4GalB1,4GIcB1-R

GbOse4Cer GalNAcB1,3Gala1,4Gal81,4GIcB1-R

GD1a:

(IV3Neu5AclI3Neu

5AcGg4Cer) Neu5Aca2,3GalB1,3GalNAcB1, 4(Neu5Aca2,3)GalB1,4GIcB1-R GD1b:

(I13(NeubAc)2Gg4

Cer) Galp1,3GalNAcB1, 4(Neu5Aca2,8Neu5Aca2,3)GalB1, 4GIcB1-R

11

GD1b-lactone: II3[Neu5Ac-(2-8,1-9)-Neu5Ac]Gg4-R LU101096 GD1c:

(IV3(NeubAc)2Gg4

Cer) NeuSAca2,8Neu5Aca2,3GalB1, 3GalNAcB1,4GalB1,4GIcB1-R GD1a :

(IV3Neu5AclIll6Neu5AcGg4Cer) Neu5Ac a2,3GalB1, 3(Neu5Aca2,6)GalNAcB1, 4GalB1,4GIcB1-R GD2:

(I13(Neu5SAc)2Gg3

Cer) GalNAcB1, 4(Neu5Aca2,8Neu5Aca2,3)GalB1, 4GIcB1-R GD3:

(II3(Neu5Ac)2Lac

Cer) Neu5Aca2,8Neu5Aca2,3GalB1, 4GIcB1-R Neu5Ac(a 2-3)DGalp(R1-1)-R pers GalNAcR1,3Gala1,4GalR1,4GlcR -R

Globo-H Fuc a 2GalR3GalNAcB3Gal a 4GalB4GIcB1-R Gal( R1- 3)GaiNAc( R 1-3)Gal( a 1- 4)Gal( B 1-4)Glc-R GalB3GalNAcB3Gala4GalB4GIicB1-Rmonosialyl-Gb5 SAa3Galf3GalNAcB3Gala4GalB4GIcB1-Rdisialyl-Gb5 SAa3GalB3GalNAcB3(SAa2-3)Gala4GalB4GIcB1-R Gala3GalB4GIcB1-R GalNAcB3Gala3GalB4GIcB1-R GalNAca3GalNAcB3Gala4GalB4GicB1-R

GM1a:r

(II3Neu5AcGg4Cer

) Galp1,3GalNAcB1, 4(Neu5Aca2,3)Galf1, 4GIc81-R Em Neu5Aca2,3GalB1,3GalNAcB1, 4GalB1,4GIcB1-Rca2,3GalB1,3GalNAcB1, 4GalB1,4GlcB1- (IV3Neu5AcGg4c |" B B ß B 12er) LU101096 GM2:

(II3Neu5AcGg3Cer

) GailNAcB1,4(Neu5Aca2,3)GalB1, 4GIcB1-R

GM2b Neub5Ac(a 2-8)Neu5Ac(a 2-3)DGalp(R1-4)DGIcp(B1-1)-R

GM3:

(II3Neu5AcLacCer

) Neu5SAca2,3GalB1,4GIcB1-R

GM4:

(I3Neu5AcaGalCer

) Neu5Aca2-3GalB1-R

GP1c:

(IV3(Neu5Ac)21I13( | Neu5Aca2,8Neu5Aca2,3GalB1, 3GalNAcB1,4(Neu5Aca2,8Neu5Aca2,

Neu5Ac)3Gg4Cer) | 8Neu5Aca2,3)GalB1,4GIcB1-R

GP1ca:

(IV3Neu5AclIll6Neu5AC,

13(Neu5Ac)3Gg4C | Neu5Ac a2,3GalB1, 3(Neu5Aca2,6)GalNAcB1,

er) 4(Neu5Aca2,8Neu5Aca2, 8Neu5Aca2,3)Gal81,4GIcB1-R

GQ1b:

(IV3(Neu5Ac)2113(_| Neu5Aca2,8Neu5Aca2,3GalB1, 3GalNAcB1,

Neu5Ac)2Gg4Cer) | 4(Neu5Aca2,8Neu5Aca2,3)GalB1, 4GIcB1-R

GQ1ba:

(IV3(Neu5Ac)21il6( | Neu5Ac a2,3GalB1, 3(Neu5Aca2,6)GalNAcB1,

Neu5Ac)2Gg4Cer) | 4(Neu5Aca2,8Neu5Aca2,3)GalB1, 4GIcB1-R

GQ1c:

(IV3Neu5AclI3(Ne | Neu5Aca2,3GalB1,3GalNAcB1, 4(Neu5Aca2,8Neu5Aca2,

u5Ac)3Gg4Cer) 8Neu5Aca2,3)GalB1, 4GIcB1-R

GT1.6T1b Neu5Ac(a 2-3)DGalp(R1-3)DGalNAc(B1-4)[NeuSAc(a 2-8)Neu5Ac(a 2-

3)]DGalp(R1-4)DGlcp(R1-1)-R GT1a: N(V3(Neu5Ac)21i3 | Neu5Aca2,8Neu5Aca2,3GalB1, 3GalNAcB1,4(Neu5Aca2,3)GalB1, 13

Neu5AcGg4Cer) |4GIcB1-R LU101096 GT1b:

(IV3NeubAcli3(Ne | Neu5SAca2,3GalB1,3GalNAcB1, 4(Neu5Aca2,8Neu5Aca2,3) u5Ac)2Gg4Cer) GalB1,4GIcB1-R

GT1c:

(I13(Neu5Ac)3Gg4 | GalB1,3GalNAcB1, 4(Neu5Aca2,8Neu5Aca2, 8Neu5Aca2,3)GalB1, Cer) 4GIcB1-R

GT1a:

(IV3Neu5AclIll6(Ne | NeuSAc a2,3GalB1, 3(Neu5Aca2,6)GalNAcB1,

u5Ac)2Gg4Cer) 4(Neu5Aca2,3)Gail81,4GIc81-R GalNAcB1,4(Neu5Aca2,8Neu5Aca2, 8Neu5Aca2,3)GalB1, 4GIcB1-R GT3:

(H3(NeuAc)3LacCer) Neu5Aca2,8Neu5Aca2,8Neu5Aca2, 3GalB1,4GIcB1-R

Gal(b1-4)GIcNAc(b1-3)Gal(b1-4)[Fuc(a1-3)}GIcNAc(b1-3)Gal(b1-

Internal Lewis x 4)GIcNAc(b1-3)Gal(b1-4)GIcNAc(b1-3)Gal(b1-4)Glc(b1-1)-R Isoglobo GalNAcB1,3Gala1,3GalR1,4GlicB - R i Galf1,3GIcNAcB1,3GalB1,4GlcB - R Gal(R1-3)[Fuc(a 1-4)]GIcNAc(R1-)-R Fuc(a 1-2)Gal(R1-3)[Fuc(a 1-4)]GIcNAc(31-)-R Gal(R1-4)[Fuc(a 1-3)]GIcNAc(R1-)-R Fuc(a 1-2)Gal(81-4)[Fuc(a 1-3)]GIcNAc(81-)-R Mole Fuc a 1,4GIcNAcR1,2Man a 1,3ManR 1,4GIcR-R GalR1,3GalR1,4GalR1,4GIcR - R

N-Acetyl GD3 Neu5Ac(a 2-8)Neu5Ac(a 2-3)Gal(b1-4)Glc(b1-1)-R

Galß 1,6GalB 1,6Galß -R

GalB1,4GIcNAcB1,3Galp1,4GlcB- R

14

NeuSAc(a2- LU101096 3)Gal(b1) Neu5Ac(a 2-3)Gal(R1-)-R Neu5Ac(a2- 3)Gal(b1- 3)GalNAc Neu5Ac(a 2-3)Gal(R1-3)GalNAc-R Neu5Ac(a2- 8)Neu5Ac(a2- 3)Gal Neu5Ac(a 2-8)Neu5Ac(a 2-3)Gal-R Neu5Aca2-8Neu5Aca2-8Neu5Aca-R Neu5Aca2-3GalB1-4GIcB-R N-Glycolyl GM3 Neu5Gc(a 2-3)Gal(R1-4)Gic(B1-1)-R NOR1 Gal(a 1- 4)GalNAc( B 1-3)Gal(a a 1- 4)Gal( B 1- 4)Glc-R Gal(a 1- 4)GalNAc- ( B 1-3)Gal(a 1- 4)GalNAc( B 1-3)Gal(a 1- 4)Gal( NOR2 ß 1-4)Glc-R NOT int GalNAc( BR 1-3)Gal(a 1-4)GalNAc(R1-3)Gal- (a 1-4)Gal( B 1- 4)Glc -R Fuc (a 1-2)Gal(R1-3)GIcNAc(B1-3)Gal-R, Neu5Ac(a 2-3)DGalp(R1-3)DGalNAc(R1-4)XNeuSAc9Ac(a 2- OAc-GT1b 8)Neu5Ac(a 2-3)]JDGalp(R-4)DGIcp(B1-1)-R P antigen (Gb4) Gal(a 1- 4)Gal- ( B 1- 4)GIcNAc( R 1-3)Gal( B 1- 4)Glc -R Pk Antigen (Gb3) | Gal(a 1-4)Gal(R1-4)GIc-R A) Gala1-4GalB1-4GIcNAcB-R GalNAcR1,4GlcR-R Sialyl Lewis a, Neu5Ac(a 2-3)Gal(R1-3)[Fuc(a 1-4)]GIcNAc(B1-)-R Sialyl Lewis c Neu5Ac(a 2-3)Gal(R1-3)GIcNAc(B1-)-R | | Neu5Ac(a 2-3)Gal(R1-4)[Fuc(a 1-3)]GIcNAc(B1-3)Gal(81-4)GIcNAc(B1- Sialyl Lewis x, 3)Gal(B1-)-R Sialyl Lewis x, Neu5Ac(a 2-3)Gal(R1-4)[Fuc(a 1-3)]GIcNAc(B1-)-R Sialyl Lewis x1 | Neu5Ac(a 2-3)Gal(81-4)[Fuc(a 1-3)]GIcNAc(81-3)Gal(81-4)GIcNAc(81-

3)Gal(R1-)-R LU101096 Trifucosyl-Lewis b | Fuc(a 1-2)Gal(b1-3)[Fuc(a 1-4)]GIcNAc(b1-3)Gal(b1-3)[Fuc(a 1- Trifucosyl-Lewis y | Fuc(a 1-2)Gal(b1-4)[Fuc(a 1-3)]GIcNAc(b1-3)Gal(b1-4)[Fuc(a 1- 16

4)Glc--R, LU101096 NeuSAc(a 2-3)Gal(31-4)GIcNAc(R1-3)Gal(R1-4)[Fuc(a -3)]GIcNAc(R1-)- VIM-2 R GalNAc(a1-3)[Fuc(a 1-2)]Gal(R1-3)GalNAc(B1-3)Gal(a 1-4)Gal(B 1- Type 4 A 4)Glc--R, Mana1-3(Mana1-6)Mana-? 3'SLN Neu5Aca2-3GalB1-4GIcNAcGB-R Neu5Aca2-6GalB1-4GIcB-R wherein R is one or more of the following: a carbohydrate/s, a peptide/s, a lipid/s, a linker/s and a chemical compound/s or substance/s or molecule/s; or R comprises one or more of the following: a carbohydrate/s, a peptide/s, a lipid/s, a linker/s or a chemical compound/s or substance/s or molecule/s; further, any one of lipid carriers as defined herein further coupled to TF disaccharide, Core-1 structure, Tn monosaccharide, Sialyl-TF mono- or disialylated, Sialyl-Tn, Polysialic acid, or mannose-6-phosphate moiety; any one of lipid carriers as defined herein further coupled to N-Glycan or O-glycan moiety (e.g., as can be found on glycoproteins or glycopeptides); any one of lipid carriers as defined herein further coupled to a carbohydrate moiety; a truncated or elongated derivative of any one of lipid carriers as defined herein; a phosphorylated, sulfated or acetylated derivative of any one of lipid carriers as defined herein; derivatives and analogues of any of any one of lipid carriers as defined herein.

[0046] As used herein, the term “glycerides” may refer to esters of glycerol (propane-1,2,3- triol) with fatty acids, widely distributed in nature. They are by long-established custom subdivided into triglycerides, 1,2- or 1,3-diglycerides, and 1- or 2-monoglycerides, according to the number and position of acyl groups (not, as one might suppose, the number of glycerol residues). The recommended method for naming individual glycerides is mono-, di- or tri-O- acylglycerol, as appropriate.

[0047] As used herein, the term “ceramide-like glycolipid” (used interchangeably with “ceramide-like glycolipid moiety”) may refer to a ceramide derivative comprising a ceramide moiety and a saccharide moiety or a B- or a-linked sacharide derivative of ceramide or analog thereof (cf. WO2010081026), wherein said B- or a-linked sacharide derivative of ceramide or analog thereof may comprise Formula I: 17

ST LU101096 R4 NH os OH wherein R4 is a linear or branched C1-C80 (e.g., C1-C36, C1-C27, C2-C36, C3-C36, C4-C36, or C16-C24) alkane or C2-C80 (e.g., C2-C36, C2-C27, C3-C36, C4-C36 or C16-C24) alkene; or R1 is -C(OH)-R3, wherein R3 is a linear or branched C1- C80 (e.g., C1-C26, C1-C36, C2-C36, or C16-C24) alkane or C2-C80 (e.g., C2-C36, C2-C26 or C16-C24) alkene; or R1 is a C6-C80 (e.g., C6-C36, C6-C27 or C16-C24) alkane or alkene, wherein: (i) C6-C80 (e.g., C6-C36, C6-C27 or C16-C24) alkane or alkene is substituted with a C5-C15 cycloalkane, C5-C15 cycloalkene, heterocycle, or aromatic ring; or (ii) C6-C80 (e.g., C6-C36, C6-C27 or C16-C24) alkane or alkene includes, within the C6- C80 alkyl or alkenyl chain, a C5-C15 cycloalkane, C5-C15 cycloalkene, heterocycle, or aromatic ring; R2 is one of the following: -CHxCH;)xCH;, -CH(OH)(CH.)xCH3, -CH(OH) CH(OH)(CH2)xCH3, -CH(OH)(CHz)xCH(CH3)2, -CH=CH(CH2)xCHz, -CH(OH)(CH2)>CH(CH3)CH2CHz, wherein X is an integer ranging from 4-17; R4 is an a-linked or a B-linked disacharide or polysacharide moiety (e.g., comprising 3 or more saccharide moieties (e.g., 3 to 5 sacharide moieties (e.g., as in GM1), e.g, comprising one or more repeat/s of the same disacharide or polysacharide moiety, preferably comprising 3 to 100 sacharide moieties, further preferably comprising 3 to 30 sacharide moieties, most preferably comprising 3 to 12 sacharide moieties), or when R1 is a linear or branched C1- C80 (e.g., C1-C36, C1-C27, C2-C36, C3-C36, C4-C36, or C16-C24) alkane, R4 is:

OH

OH oO

OH OH OH A

OH OH © and Ais O or -CHy; In a preferred embodiment, the R4 (e.g., an epitope, e.g., an epitope for a toxin) may comprise 3 to 5 sacharide moieties (i.e. sugars, e.g., as in GM1), wherein the binding capacity of the lipid carrier of the present invention may be optionally increased in that the 18polysaccharide moiety may comprise several repeats of the same R4 (e.g., epitope), wherein LU101096 said R4 may comprise more than 12 saccahride moities.

[0048] As used herein, the term “ceramide” (or “ceramide moiety”) may refer to compound or substance composed of sphingosine and a fatty acid.

[0049] As used herein, the term “sphingolipid” may refer to a class of lipids containing a backbone of sphingoid bases and a set of aliphatic amino alcohols that includes sphingosine or a substance structurally similar to it. Non-limiting examples of sphingolipids include: sphingosine, sphingomyelins, cerebroside, sulfatides, globosides, and gangliosides.

[0050] As used herein, the term “sphingosine” (or “sphingosine moiety”) may refer to 2- amino-4-trans-octadecene-1,3-diol), which is an 18-carbon amino alcohol with an unsaturated hydrocarbon chain, which forms a primary part of sphingolipids, a class of cell membrane lipids that include sphingomyelin, an important phospholipid.

[0051] As used herein, the term “ganglioside” may refer to a molecule composed of a glycosphingolipid (i.e., a subtype of glycolipids containing the amino alcohol sphingosine, e.g. ceramide and oligosaccharide) with one or more sialic acids (e.g., n-acetylneuraminic acid, NANA) linked on the sugar chain.

[0052] As used herein, the term “fatty acid” (used interchangeably with “fatty acid moiety”) may refer to aliphatic monocarboxylic acids derived from or contained in esterified form in an animal or vegetable fat, oil or wax. Natural fatty acids commonly have a chain of 4 to 28 carbons (usually unbranched and even-numbered), which may be saturated or unsaturated. By extension, the term also embraces all acyclic aliphatic carboxylic acids and very long chains over 28 carbons (up to 80).

[0053] As used herein, the term “phospholipids” may refer to lipids containing phosphoric acid as mono- or di-esters, including phosphatidic acids and phosphoglycerides.

[0054] As used herein, the term “steroid” (used interchangeably with “steroid moiety”) may refer to naturally occurring compounds and synthetic analogues, based on the cyclopenta[a]phenanthrene carbon skeleton, partially or completely hydrogenated; there are usually methy! groups at C-10 and C-13, and often an alkyl group at C-17. By extension, one or more bond scissions, ring expansions and/or ring contractions of the skeleton may have occurred.

[0055] As used herein, the term ,lipopeptide” may refer to a molecule comprising a peptide part (e.g., a peptide molecule as described herein) associated with (e.g. coupled to) a lipid part (e.g., a lipid molecule as described herein). Preferably said peptide part of the lipopeptide is linked directly to said lipid part of the lipopeptide via a covalent bond or a linker or linking molecule (e.g., a peptide-based linker or a chemical linker such as e.g. bi-functional linkers (NHS-ester and maleimid), copper-free click-chemistry alkyne-azido triazole linkages, unnatural amino acids, carbohydrate-mediate linkages, photocross-linkers. Other non- 19covalent linkages allowing an association (e.g., a stable association) between said peptide LU101096 part and said lipid part are also within the scope of the present invention. The peptide of the peptide part can be of any length and may include one or more modifications, preferably post-translational modifications, including, but not limited to, glycosylation, sulfation, carboxylation, phosphorylation etc. The peptide part can also be a protein composed of one or more covalently or non-covalently associated polypeptide chains. The peptide can be covalently or non-covalently coupled to an additional chemical molecule mainly for stabilizing a certain conformation of the peptide, such as in bicyclic peptides [Zorzi A, Deyle K, Heinis C. Curr Opin Chem Biol. 2017 Jun;38:24-29. doi: 10.1016/j.cbpa.2017.02.006. Epub 2017 Feb

27. Review], whereby this coupling to an additional chemical molecule can be within one polypetide chain or between two or polypeptide chains. Each polypeptide chain may independently be modified, preferably post-translationally or chemically modified. In a preferred embodiment the peptide part is a binding molecule. Further preferably, the peptide part comprises an antibody, mono-, di, -or tri-specific, a derivative of an antibody, an antibody fragment, a single chain antibody comprising relevant parts of a VH and a VL chain, a single domain antibody, or another peptide-based molecule comprising at least one VH or VL domain, a fragment thereof or mutated or modified version binding to a toxin of the invention (e.g., any toxin described herein), a toxin receptor (e.g., any toxin receptor described herein), a receptor, a cell, a protein, an immunologically active molecule, an inflammatory molecule or another molecule on the pathogenic microorganism (e.g., any pathogenic microorganism described herein). In another preferred embodiment the peptide can be a molecule which mimicks a carbohydrate, e.g. antiidiotypic antibody, antibody fragment, scFv, single domain antibody or a peptide or bicyclic peptide. Those molecules may comprise one or more peptide-based linkers. In another preferred embodiment the peptide part comprises a lectin able to bind a carbohydrate or a fragment thereof. In another preferred embodiment the peptide part comprises an immunologically active molecule such as a cytokine or chemokine or a fragment thereof. In a preferred embodiment the peptide part binds to a toxin of the invention, a toxin receptor, a receptor, a cell, a protein, an immunologically active molecule, an inflammatory molecule or another molecule on the pathogenic microorganism. The peptide part of the lipopeptide can also be any other naturally occurring, therefrom derived, or chemically synthetized chemical moiety able to bind to a toxin of the invention, a toxin receptor, a receptor, a cell, a protein, a carbohydrate or another molecule on the pathogenic microorganism, or an immunologically active molecule, preferably causing an inflammation. In a certain embodiment such molecules comprise DNA or RNA or a DNA or RNA peptide or protein complex. In preferred embodiments bicyclic peptides may be used, which use besides the peptide part a chemical molecule which adds to conformational structure. For example, it may not only be bicyclic, but could be also tricyclic a.o.: [Cyclic peptide 20therapeutics: past, present and future. Zorzi A, Deyle K, Heinis C. Curr Opin Chem Biol. 2017 LU101096 Jun;38:24-29. doi: 10.1016/j.cbpa.2017.02.006. Epub 2017 Feb 27. Review.] peptides. In further embodiments peptide/protein based binding molecules, which are modified to keep/stabilize their binding structure in environments like GI tract, Lung, urogenital tract better than the non-modified form, are used. Modifications, e.g., mutation of sites prone for proteolysis or of sites which when mutated help to stabilize the spacial three-dimensional structure, may also be used. Technologies to achieve this are known in the art. An advantage will be the generation of phage-display libraries displaying such molecules for selection of binders to toxins and/or pathogens, which is within scope of the present invention. Various lipid molecules of different type, length of fatty acid chains, with or without natural and chemical modifications may be used in the present invention. The sequence of the peptide/protein part may be designed accordingly and tested with suitable amino acids and side chains for covalent coupling. Chemical, peptide and carbohydrate-based spacers may be used for a better presentation on the bacteria. The coupling of the peptides with lipids can be for example carried on by the use of bi-functional linkers (e.g., NHS-ester and maleimid), copper-free click-chemistry alkyne-azido triazole linkages, unnatural amino acids, carbohydrate-mediate linkages, photocross-linkers a.o., which readily known in the art.

[0056] As used herein, the term “peptidomimetic compound” includes a peptide, which mimics a biological effect or chemical structure of another peptide, carbohydrate or a chemical entity; as well as a chemical entity, which mimics a peptide- or protein structure, epitope or determinant.

[0057] As used herein, the term “receptor” or “toxin receptor’ may refer to a toxin-binding molecule present on the surface of a cell, preferably of a mammalian cell. It mayrefer to a molecule present on the surface of a cell, preferably of a mammalian cell, which enables or facilitates the binding of a pathogenic microorganism or pathogen to said cell. Cells of interest include, for example epithelial or endothelial cells, in particular those that are part of a mammalian mucosal membrane or epithelia, such as human or animal mucosal membranes. Toxins for which receptors are described include but are not limited to Shiga toxin Stxl, Stx2, Stx2c, Stx2d, Stx2e, Stx2f, C. difficile toxin A, C. difficile toxin B, C. botulinum toxin, Vibrio cholera toxin, E. coli heat labile enterotoxin Type 1, Escherichia coli heat-stable enterotoxin, Clostridium perfringens enterotoxin. Such toxin receptors include, but are not limited to, GUCY2C (guanylate cyclase 2C (heat stable enterotoxin receptor)), Heat labile enterotoxin- and Cholera Toxin-GM1 Ganglioside Receptor, Clostridium perfringens enterotoxin-receptors Claudin-3 and Claudin-4, Clostridium difficile A and B- receptors Combined Repetitive OligoPeptides (CROP'S), Clostridium difficile A receptor ß- type trisaccharide œGal(1,3)-B Gal(1,4)-B GIcNAc, Shiga toxin Stxl, Stx2, Stx2c, Stx2d 21

Glycolipid receptor Globotriaosyl ceramide (Gbs) or Shiga toxin Stx2e Glycolipid receptor | 101096 Globotetraosyl ceramide (Gb41).

[0058] As used herein, the term “carbohydrate” (used interchangeably with “carbohydrate moiety”) may refer to compounds such as aldoses and ketoses having the stoichiometric formula Cn(H20)n (e.g., hence “hydrates of carbon”). The generic term “carbohydrate” includes, but is not limited to, monosaccharides, oligosaccharides and polysaccharides as well as substances derived from monosaccharides by reduction of the carbonyl group (alditols), by oxidation of one or more terminal groups to carboxylic acids, or by replacement of one or more hydroxy group(s) by a hydrogen atom, an amino group, thiol group or similar groups. It also includes derivatives of these compounds.

[0059] As used herein, the term "toxin" includes toxins in their naturally occurring form, inactivated toxins and fragments or derivatives of toxins such as recombinant toxins of a pathogenic microorganism, for example pathogenic microorganisms. Toxins in connection with present invention are preferably toxins which are relevant for endangering health and/or well-being of humans or non-human animals, such cattle, pig, horse, sheep, goat, cats, dogs, ducks, goose, chicken, fish, etc. A toxin is preferably a toxin produced either by a bacterium belonging to a family selected from the group consisting of Enterobacteriaceae, Clostridiaceae, Vibrionaceae, Staphylococcaceae, Streptococcaceae, Helicobacteraceae, Pseudomonadaceae, Pasteurellaceae, Chlamydiaceae, Campylobacteraceae, Aeromonadaceae, Neisseriaceae, Listeriaceae, Corynebacteriaceae, Aeromonadales, Bacteroidaceae, Bordetella, Bacillaceae or a protozoa belonging to a family selected from the group consisting of Acanthamoebidae, Amoebida, Hexamitidae, Cryptosporidiidae or a fungi belonging to a family selected from the group consisting of Saccharomycetaceae, Trichocomaceae, Clavicipitaceae, Nectriaceae. Furthermore "toxin" may refer herein preferably to a toxin produced either by a bacterium belonging to a genus selected from the group consisting of Enterobacter, Echerischia, Shigella, Clostridium, Vibrio, Staphylococcus, Streptococcus, Helicobacter, Pseudomonas, Haemophilus, Chlamydia, Campylobacter, Salmonella, Citrobacter, Yersinia, Pasteurella, Neisseria, Listeria, Corynebacterium, Klebsiella, Aeromonas, Serratia, Proteus, Bacteroides, Bordetella, Bacillus or a protozoa belonging to a genus selected from the group consisting of Acanthamoeba, Entamoeba, Giardia, Cryptosporidium or a fungi belonging to a genus selected from the group consisting of Candida, Penicillium, Aspergillus, Claviceps, Paecilomyces, Fusarium. Furthermore, toxin includes toxins made in the gut. Preferably "toxin" may refer to an enterotoxin produced by a pathogenic microorganism. Within the context of the present invention the term "toxin" includes, but is not limited to toxins of the following list: E. coli: Heat labile toxin (LT), Heat stabile toxin (ST), VerotoxinsIShiga like toxins (Stxs), Cytotoxins, endotoxins (LPS), EnteroAggregative ST toxin (EAST), Shigella: Shiga toxin (STxs), Shigella enterotoxins 1 22

(ShETI), Shigella enterotoxins 2 (ShET2), Neurotoxin; Salmonella: Cytolethal distending LU101096 toxins (Cat), AvrA toxin; Yersinia: Cytotoxic necrotizing facto (CNFy), Yersinia murine toxin (Ymt), Yst toxin, Toxin complex (TCa), Heat stabile toxin; Enterobacter: E. cloacae leukotoxin, Shiga-like toxin Il, Klebsiella: heat-stable like enterotoxins, extracellular toxic complex (ETC); Serratia: Hemolysins (Shi), Pore-forming Toxin (PFT), Proteus: a-hemolysin (HiyA), Citrobacter: heat-stable like toxin, Cytotoxins; Clostridium: C. perfringens alpha-toxin (CpPLC), C. perfringens beta toxin, C. perfringens enterotoxin (CPE), C. difficile enterotoxins (Ted), C. butulinum Neurotoxins, C. tetani Tetanospasmin, C. butulinum C2 toxin, C. butulinum C3 toxin, C. perfringens epsilon-toxin (etoxin), C. perfringens iota-toxin (L-toxin), tetanus neurotoxin (TeNT), theta-toxinIPFO (perfringolysin 0), C. spiroforme (spiroforme toxin), C. septicum (a-toxin), Lecithinase; Vibrio : Cholera toxins (CTx), accessory cholera enterotoxin (Ace), RTX toxin, zona occludens toxin (Zot), Cholix toxin; Staphylococcus: a- hemolysin, P-hemolysin, 6-hemolysin, y-hemolysin, Exfoliative toxins (Exofoliatins), Panton- Valentine leukocidin (PVL), staphylococcal enterotoxins (SE), Toxic shock syndrome toxin-l (TSST-1 ) Streptococcus: P-haemolysinicytolysin, CAMP factor, Streptolysin 0 , Streptolysin S, Pneumolysin, S. pyogenes Exotoxins (PSE), Helicobacter: vacuolating cytotoxin A (VacA), Cytolytic toxins Pseudomonas: Exotoxins (ex: ExoA, ExoS, ExoT, ExoU, ExoY), Phospholipase C (PLC) Pasteurella: Pasteurella Multocida Toxin (PMT), RTX toxins Bacillus: B. weihenstephanensis endotoxins, B. cereus Hemolysin BL (Hbl), B. cereus, onhemolytic Enterotoxin (Nhe), B. cereus Cytotoxin K (CytK), B. cereus emetic toxin, B. cereus toxin (Cereolysin), B. anthracis (Anthrax toxin), B. thuringiensis 6-endotoxins (Cry toxins), Campylobacter: Cytolethal distending toxin (cdtA, cdtB, cdtC), cholera-like enterotoxin Aeromonas: Aerolysin Cytotoxic Enterotoxin (ACT), ADP-ribosylation toxin, a-hemolysins, bhemolysins, Heat labile toxin (LT+), Heat stabile toxin (ST+) Neisseria: endotoxins (LPS) Bordetella : B. pertussis (pertusis toxin), Adenylate cyclase toxin, Tracheal cytotoxin, Dermonecrotic (heat-labile) toxin, endotoxins (LPS) Haemophilus: Endotoxin (LOS), Cytolethal distending toxins (HACDT), Hemolysins Chlamydia: Endotoxins Corynebacteria: Cytotoxins, Diphteria toxin, Exotoxins Bacteroides: Bacteroides fragilis toxin (bft) Listeria: Listeriolysin 0.

[0060] In a preferred embodiment, the toxin is Heat labile toxin (LT), Heat stabile toxin (ST), Verotoxinsl Shiga like toxins (Stxs), Cytotoxins, endotoxins (LPS), EnteroAggregative ST toxin (EAST), Shiga toxin (STxs), Shigella enterotoxins 1 (ShETI), Shigella enterotoxins 2 (ShET2), Neurotoxin, Cytolethal distending toxins (Cdt), AvrA toxin, Cytotoxic necrotizing factor (CNFy), Yersinia murine toxin (Ymt), Yst toxin, Toxin complex (TCa), Heat stabile toxin, E. cloacae leukotoxin, Shiga-like toxin li, heat-stable like enterotoxins, extracellular toxic complex (ETC), Hemolysins (Shi), Pore-forming Toxin (PFT), a-hemolysin (HIyA), heat- stable like toxin, Cytotoxins, C. perfringens alpha-toxin (CpPLC), C. perfringens beta toxin, 23

C. perfringens enterotoxin (CPE), C. difficile enterotoxins (Tcd), C. butulinum Neurotoxins, C. LU101096 tetani Tetanospasmin, C. butulinum C2 toxin , C. butulinum C3 toxin, C. perfringens epsilontoxin (etoxin), C. perfringens iota-toxin (L-toxin), tetanus neurotoxin (TeNT), theta- toxinIPFO (perfringolysin 0), C. spiroforme (spiroforme toxin), C. septicum (a-toxin), Lecithinase, Cholera toxins (CTx), accessory cholera enterotoxin (Ace), RTX toxin, zona occludens toxin (Zot), Cholix toxin, a-hemolysin, P-hemolysin, 6-hemolysin, y-hemolysin, Exfoliative toxins (Exofoliatins), Panton-Valentine leukocidin (PVL), staphylococcal enterotoxins (SE), Toxic shock syndrome toxin-l (TSST-l), P-haemolysinicytolysin, CAMP factor, Streptolysin 0 , Streptolysin S, Pneumolysin, S. pyogenes Exotoxins (PSE), vacuolating cytotoxin A (VacA), Cytolytic toxins, Exotoxins (ex : ExoÂ, ExoS, ExoT, ExoU, ExoY), Phospholipase C (PLC), Pasteurella Multocida Toxin (PMT), RTX toxins, B. weihenstephanensis endotoxins, B. cereus Hemolysin BL (Hbl), B. cereus , onhemolytic Enterotoxin (Nhe), B. cereus Cytotoxin K (CytK), B. cereus emetic toxin, B. cereus toxin (Cereolysin), B. anthracis (Anthrax toxin), B. thuringiensis 6-endotoxins (Cry toxins), Cytolethal distending toxin (cdtA, cdtB, cdtC), cholera-like enterotoxin, Aerolysin Cytotoxic Enterotoxin (ACT), ADP-ribosylation toxin, ahemolysins, b-hemolysins, Heat labile toxin (LT+), Heat stabile toxin (ST+), endotoxins (LPS), B. pertussis (pertusis toxin), Adenylate cyclase toxin, Tracheal cytotoxin, Dermonecrotic (heat-labile) toxin, endotoxins (LPS), Endotoxin (LOS), Cytolethal distending toxins (HdCDT), Hemolysins, Endotoxins, Cytotoxins, Diphteria toxin, Exotoxins, Bacteroides fragilis toxin (bft), Listeriolysin 0, or rota virus toxin (NSP4).

[0061] As used herein, the term “mammalian” may refer to a mammal organism, e.g., such as human or an animal, such as a dog, cat, cattle, pig, horse, camel, sheep, mouse, rat, poultry, fish preferably human.

[0062] An “antibody” when used herein is a protein comprising one or more polypeptides (comprising one or more binding domains, preferably antigen binding domains) substantially or partially encoded by immunoglobulin genes or fragments of immunoglobulin genes. The term "immunoglobulin" (lg) is used interchangeably with "antibody" herein. The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as myriad immunoglobulin variable region genes. In particular, an “antibody” when used herein, is typically tetrameric glycosylated proteins composed of two light (L) chains of approximately 25 kDa each and two heavy (H) chains of approximately 50 kDa each. Two types of light chain, termed lambda and kappa, may be found in antibodies. Depending on the amino acid sequence of the constant domain of heavy chains, immunoglobulins can be assigned to five major classes: A, D, E, G, and M, and several of these may be further divided into subclasses (isotypes), e.g., IgG1, 1gG2, 1gG3, 19G4, IgA1, and IgA2, with IgG being preferred in the context of the present invention. An antibody of the 24present invention is also envisaged which has an IgE constant domain or portion thereof that LU101096 is bound by the Fc epsilon receptor|. An IgM antibody consists of 5 of the basic heterotetramer unit along with an additional polypeptide called a J chain, and contains 10 antigen binding sites, while IgA antibodies comprise from 2-5 of the basic 4-chain units which can polymerize to form polyvalent assemblages in combination with the J chain. In the case of IgGs, the 4-chain unit is generally about 150,000 daltons. Each light chain includes an N- terminal variable (V) domain (VL) and a constant (C) domain (CL). Each heavy chain includes an N-terminal V domain (VH), three or four C domains (CHs), and a hinge region. The constant domains are not involved directly in binding an antibody to an antigen, but can exhibit various effector functions, such as participation of the antibody dependent cellular cytotoxicity (ADCC). If an antibody should exert ADCC, it is preferably of the 19G1 subtype, while the IgG4 subtype would not have the capability to exert ADCC. The term “antibody” also includes, but is not limited to, but encompasses monoclonal, monospecific, poly- or multi-specific antibodies such as bispecific antibodies, humanized, camelized, human, single- chain, chimeric, synthetic, recombinant, hybrid, mutated, grafted, and in vitro generated antibodies, with chimeric or humanized antibodies being preferred. The term "humanized antibody" is commonly defined for an antibody in which the specificity encoding CDRs of HC and LC have been transferred to an appropriate human variable frameworks ("CDR grafting"). The term “antibody” also includes scFvs, single chain antibodies, diabodies or tetrabodies, domain antibodies (dAbs) and nanobodies and parts thereof able to bind to a toxin or bacterial receptor. In terms of the present invention, the term “antibody” shall also comprise bi-, tri- or multimeric or bi-, tri- or multifunctional antibodies having several antigen binding sites.

[0063] Furthermore, the term "antibody" as employed in the invention also relates to derivatives of the antibodies (including fragments) described herein. A "derivative" of an antibody comprises an amino acid sequence which has been altered by the introduction of amino acid residue substitutions, deletions or additions. Additionally, a derivative encompasses antibodies which have been modified by a covalent attachment of a molecule of any type to the antibody or protein. Examples of such molecules include sugars, PEG, hydroxyl-, ethoxy-, carboxy- or amine-groups but are not limited to these. In effect the covalent modifications of the antibodies lead to the glycosylation, pegylation, acetylation, phosphorylation, amidation, without being limited to these. In a preferred embodiment the amino acid sequence is mutated to stabilize the protein part of the lipopeptide especially at its site of action in the mammalian, such as the gastrointestinal system or parts of it.

[0064] As used herein, the terms "effective amount” or "therapeutically effective amount,” refer to an amount of an active agent (e.g., modified microorganism) as described herein that is sufficient to achieve, or contribute towards achieving, one or more desirable clinical 25outcomes. An appropriate "effective" amount in any individual case may he determined using LU101096 standard techniques known in the art, such as a dose escalation study.

[0065] As used herein, “about” means especially +/-10%, +/-5% or +/-3% (referring to the given numeric value), if not indicated otherwise. In each of the invention embodiments, “about” can be deleted.

[0066] As used herein, “administering” a compound (or microorganism) can be affected or performed using any of the various methods and delivery systems known to those skilled in the art. The administering can be performed, for example, intravenously, orally, nasally, via the cerebrospinal fluid, via implant, transmucosally, transdermally, intramuscularly, intraocularly, parenterally, topically, via inhalation and subcutaneously. The following delivery systems, which employ a number of routinely used pharmaceutically acceptable carriers, are only representative of the many embodiments envisioned for administering compositions according to the instant methods.

[0067] The appropriate dosage, or therapeutically effective amount, will depend on the condition to be treated, the severity of the condition, prior therapy, and the patient's clinical history and response to the therapeutic agent. The proper dose can be adjusted according to the judgment of the attending physician such that it can be administered to the patient one time or over a series of administrations. The pharmaceutical composition can be administered as a sole therapeutic or in combination with additional therapies as needed.

[0068] If the pharmaceutical composition has been lyophilized, the lyophilized material is first reconstituted in an appropriate liquid prior to administration. The lyophilized material may be reconstituted in, e.g., bacteriostatic water for injection (BWFI), physiological saline, phosphate buffered saline (PBS), or the same formulation the protein had been in prior to lyophilization. However, a modified microorganism of the present invention (e.g., lyophilized bacteria) could also be encapsulated and directly administred orally (e.g., without being reconstituted).

[0069] Pharmaceutical compositions for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. In addition, several recent drug delivery approaches have been developed and the pharmaceutical compositions of the present invention are suitable for administration using these new methods, e. g., Inject- ease, Genject, injector pens, and needleless devices such as MediJector and BioJector. The present pharmaceutical composition can also be adapted for yet to be discovered administration methods. See also Langer, 1990, Science, 249: 1527-1533.

[0070] The pharmaceutical composition can also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously, into the ligament or tendon, subsynovially or intramuscularly), by subsynovial injection or by intramuscular injection. Thus, for example, the formulations may be modified with suitable 26polymeric or hydrophobic materials (for example as a emulsion in an acceptable oil) or ion Lu101096 exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble sait.

[0071] The pharmaceutical compositions may also be in a variety of conventional depot forms employed for administration to provide reactive compositions. These include, for example, solid, semi-solid and liquid dosage forms, such as liquid solutions or suspensions, slurries, gels, creams, balms, emulsions, lotions, powders, sprays, foams, pastes, ointments, salves, balms and drops.

[0072] The pharmaceutical compositions may, if desired, be presented in a vial, pack or dispenser device which may contain one or more units dosage forms containing the active ingredient. In one embodiment, the dispenser device can comprise a syringe having a single dose of the liquid formulation ready for injection. The syringe can be accompanied by instructions for administration.

[0073] The pharmaceutical composition may further comprise additional pharmaceutically acceptable components. Other pharmaceutically acceptable carriers, excipients, or stabilizers, such as those described in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980) may also be included in formulations described herein, provided that they do not adversely affect the desired characteristics of the formulation. As used herein, "pharmaceutically acceptable carrier" means any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed and include: additional buffering agents; preservatives, co-solvents; antioxidants, including ascorbic acid and methionine; chelating agents such as EDTA; metal complexes (e.g., Zn- protein complexes); biodegradable polymers, such as polyesters; salt-forming counterions, such as sodium, polyhydric sugar alcohols; amino acids, such as alanine, glycine, asparagine, 2-phenylalanine and threonine; sugars or sugar alcohols, such as lactitol, stachyose, mannose, sorbose, xylose, ribose, ribitol, myoinisitose, myoinisitol, galactose, galactitol, glycerol, cyclitols (e.g., inositol), polyethylene glycol; sulfur containing reducing agents, such as glutathione, thioctic acid, sodium thioglycolate, thioglycerol, [alpha]- monothioglycerol, and sodium thio sulfate; low molecular weight proteins, such as human serum albumin, bovine serum albumin, gelatin, or other immunoglobulins; and hydrophilic polymers, such as polyvinylpyrrolidone.

[0074] The formulations described herein are useful as pharmaceutical compositions in the treatment and/or prevention of the pathological medical condition as described herein in a subject (e.g., a patient in need thereof, e.g., in a healtly subject for preventative purposes). The term "treatment" refers to both therapeutic treatment and prophylactic or preventative 27measures. Treatment includes the application or administration of the formulation to the body, LU101096 an isolated tissue, or cell from a patient who has a disease/disorder, a symptom of a disease/disorder, or a predisposition toward a disease/disorder, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve, or affect the disease, the symptom of the disease, or the predisposition toward the disease.

[0075] The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like.

[0076] The compositions of the invention can be formulated as neutral or salt forms.

[0077] Pharmaceutically acceptable salts include, but are not limited to those formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.

[0078] As used herein, the term "treating" and "treatment" refers to administering to a subject a therapeutically effective amount of a pharmaceutical composition according to the invention. A “therapeutically effective amount” may refer to an amount of the pharmaceutical composition which is sufficient to treat or ameliorate a disease or disorder, to delay the onset of a disease or to provide any therapeutical benefit in the treatment or management of a disease.

[0079] As used herein, the term “prophylaxis” refers to the use of an agent for the prevention of the onset of a disease, disorder, syndrome or condition. A “prophylactically effective amount” defines an amount of the active component or pharmaceutical agent sufficient to prevent the onset or recurrence of a disease or to prevent or alleviate symptomes and/or complications related to the disease.

[0080] The term “polypeptide” is equally used herein with the term "protein". Proteins (including fragments thereof, preferably biologically active fragments, and peptides, usually having less than 30 amino acids) comprise one or more amino acids coupled to each other via a covalent peptide bond (resulting in a chain of amino acids). The term "polypeptide" as used herein describes a group of molecules, which, for example, consist of more than 30 amino acids. Polypeptides may further form multimers such as dimers, trimers and higher oligomers, i.e. consisting of more than one polypeptide molecule. Polypeptide molecules forming such dimers, trimers etc. may be identical or non-identical. The corresponding higher order structures of such multimers are, consequently, termed homo- or heterodimers, homo- or heterotrimers etc. The terms "polypeptide" and "protein" also refer to naturally modified | polypeptides/proteins wherein the modification is effected e.g. by post-translational 28

DImodifications like glycosylation, acetylation, phosphorylation and the like. Such modifications LU101096 are well known in the art.

[0081] The term "subject" is intended to include living organisms. Examples of subjects include mammals, e.g., humans, dogs, cows, horses, pigs, sheep, goats, cats, mice, rabbits, rats, and transgenic non-human animals. In preferred embodiments of the invention, the subject is a human (e.g., patient).

[0082] Those "in need of treatment” include those already with the disorder, as well as those in which the disorder is to be prevented.

[0083] Infectious Diseases:

[0084] For a number of infectious diseases, effective vaccines are missing, and the increasing rate of drug resistances is complicating the use of conventional antimicrobial therapy. Due to this there is a need for novel therapeutic and prophylactic approaches against infectious diseases, in particular enteric infectious diseases which continue to cause massive morbidity and mortality in humans. Effective vaccines are still not available for a number of important diarrheal diseases, and, as mentioned, controlling these with conventional antimicrobial therapy is being complicated by increasing rates of drug resistance.

[0085] The initial and critical step that leads to an infection is the binding of a pathogen or its toxin(s) to host cells. Anti-adhesion strategies aim to prevent and/or displace said binding and to prevent or treat the subsequent infection and/or its symptoms. Anti-adhesion strategies have attracted increasing interest as a source of novel therapeutics to prevent and treat infectious diseases. An advantage of such approaches is that the pathogen is not killed. As a consequence, anti-adhesion strategies may avoid problems associated with release of toxic products from dead bacteria and they may put much less selection pressure on pathogens, reducing the risk of resistance development.

[0086] Several anti-adhesion approaches may be envisaged, including providing receptor analogs or adhesin analogs, inhibition of adhesins and their host receptors, vaccination with adhesins or analogs, or inhibiting the synthesis of adhesins or their host receptor. Although the initial adhesion of pathogens or their toxins to host cells may happen through protein- protein interactions or phospholipid-protein interactions it is often mediated by protein- carbohydrate interactions. As a consequence, many carbohydrates or carbohydrate mimicking substances have been developed based either on proteins, polymers, calixarenes, dendrimers, cyclodextrins, cyclopeptides, fullerenes, gold nanoparticles and quantum dots. In order to efficiently block protein-carbohydrate interactions synthetic neutralization agents need to comprise multiple oligosaccharide epitopes displayed on complex three-dimensional scaffolds, conditions that may be difficult to reproduce synthetically. However, to date, clinical 29results in human were disappointing, mostly because of toxicity or lack of efficacy [e.g. LU101096 Synsorb PK (Synsorb Biotech); Tolevamer (Genzyme)].

[0087] Accordingly, US Patent 6,833,130 discloses recombinant microorganisms, genetically modified to express carbohydrate structures that mimic the natural binding moieties of bacterial toxins. Such microorganisms are able to present the binding moiety at high density. The efficacy of these microorganisms in binding toxins and protecting animals in lethal challenge models has been demonstrated. However, in order to use microorganisms in humans and animals, it is not only necessary for these microorganisms to express a binding moiety for a pathogenic ligand at sufficient density, but it should preferably be harmless, ideally it should be non-pathogenic and not genetically modified or recombinant.

[0088] There is some prior art that reports the ability of non-pathogenic microorganisms to specifically co-aggregate other microorganisms. However, the detailed mechanism of interaction is unknown. A few pathogenic microorganisms were described to naturally express structures that may mimic natural binding moiety for pathogenic ligands. Some publications further report the possibility to produce nanoparticles decoys with toxin receptor to bind the toxin. The potential of this approach is supported through in vitro and in vivo studies. However, the use of nanoparticles in the gastrointestinal tract may be limited by natural biodegradation, absorption and their potential intrinsic toxic effect on epithelial cells, on the gut microbiota or on extra-intestinal organs where they may accumulate. Those potential toxic effects may be further amplified if the nanoparticles are decoyed with a pathogen or a toxin. Furthermore, the efficacy of nanoparticles may be limited by many factors including susceptibility to gastrointestinal environment (enzymes, pH etc.) or the inability of some nanoparticles to cross the mucus layer and to locate near the epithelium where pathogen microorganisms may locally release their toxin. Thus, as is evident from the above, the prior art provides recombinant microorganisms for use in treating infectious | disease, particularly enteric infectious disease which may be harmful, and which are not be | food grade organisms. Alternatively, the prior art provides agents which have shown to be toxic when administered to mammals and for which toxicity potential is still largely unknown.

[0089] Currently, no non-pathogenic microorganisms capable of binding a lipid-carrier (i.e. glycolipid or a lipopeptide) whose carried moiety (i.e. carbohydrate or peptide) may be able to bind a toxin, the receptor for a toxin, a pathogen or the receptor for a pathogen have been described. Furthermore, no non-pathogenic microorganisms capable of binding a fusion- protein composed of an anchor-domain and a peptide that is able to bind a toxin, the receptor for a toxin, a pathogen or the receptor for a pathogen have been described. However, it is highly desirable to block the interaction between pathogenic ligands such as toxins from pathogenic microorganisms or the pathogen microorganism itself and their cognate receptors on the surface of mammalian cells, since this interaction is crucial for 30 ee ESdisease pathogenesis. Such a strategy is particularly promising for fighting against enteric Lu101096 infectious diseases which cause a high morbidity and mortality in humans, since no effective vaccines are available and rates of drug resistance against conventional antibiotic therapies increase. Due to this there is a need for novel therapeutic and prophylactic approaches against infectious diseases, in particular enteric infectious diseases. It, thus, follows that the technical problem underlying the present invention is to comply with the needs described above. The solution to this technical problem is achieved by providing the embodiments characterized herein, exemplified in the appended examples and set out in the claims.

[0090] Gastrointestinal disorders and gastrointestinal cancers:

[0091] Gastrointestinal disorders include acute, chronic, recurrent or functional disorders while covering a broad range of diseases, including the most common acute and chronic inflammatory bowel disease, Celiac Disease etc. For most of these disorders, no specific treatment is available. The primary therapy is mostly relying on immune suppressor that may have significant side effects. Other existing approaches are based on the microbiota. Thus, in severe cases, feces transplantation was successfully used.

[0092] Another approach that attracted attention is the use of non-pathogenic lactic acid bacteria with anti-inflammatory properties. Such microorganisms may be taken chronically without risk of side effects. However, this approach is still confronted to several limits.

[0093] Thus, if the anti-inflammatory properties are introduced through genetic modifications, risks related to the chronic intake of recombinant microorganisms may significantly limit their use. A more appealing approach is the use of strains with natural anti-inflammatory properties. However, the gut contains an amazing number of microorganisms (up to 1x10" per gram feces). Thus, the potential anti-inflammatory effect of exogenously administered bacteria is strongly diluted. Gastrointestinal adenocarcinoma is a common malignant disease worldwide. Non-Pathogenic microorganisms, especially Lactic-Acid Bacteria (LAB) have been reported to have anticancer properties like antioxidant activity, inhibition of tumor cell proliferation or induction of apoptosis. However, the use of non-pathogenic microorganisms may be confronted to the same limits as those mentioned above.

[0094] The beneficial effect of non-pathogenic with beneficial properties (anti-inflammatory, anti-cancer etc.) would be much stronger if such microorganisms would be able to specifically target the site of disease (site of inflammation or the tumor). Thus, there is a need for a new strategy that may help to target the non-pathogenic microorganisms with beneficial properties at the sites of need without genetic modifications. Along the intestinal tract and depending on the physiological or pathological conditions endothelial cells may express different carbohydrate structures or carbohydrate receptors. As example, under inflammation conditions (as it is the case in IBD), cells are known to express new and specific 31carbohydrate structures. Furthermore, cancer cells are known to express new carbohydrate Lu101096 antigens not present on the surface of healthy cells.

[0095] Currently, only GMO approaches or non-GMO method using peptide fused to a protein-domains know to anchor themselves to the surface of suitable non-pathogenic microorganisms are available to display specific ligand or receptor on the surface of non- pathogenic microorganisms. Thus, there is a high need for new non-GMO methods that would allow displaying a receptor or a ligand (especially when the said receptor or ligand is a carbohydrate structure) on the surface of microorganisms with beneficial properties, thereby enabling the said microorganisms to target the site where their beneficial properties are needed.

[0096] Mucosal Immunization:

[0097] The use of non-pathogenic bacteria like LAB to induce antigen specific immune responses is a field that is attracting some attention. Such approaches may be used to develop vaccines against pathogens, vaccines against cancer or allergy vaccines (that induce tolerance against the antigen). Again, to induce a non-pathogenic microorganism to display an antigen on its surface the most used approach is the genetic modification of the said non-pathogenic microorganisms. Beside the fact that this may result in the release of live recombinant microorganisms into nature, significant safety concerns have been raised that may limit the application of this technology including i) transfer of material to other microorganisms, ii) integration of the plasmid into the genome of the recipient or iii) integration of the DNA into host genome.

[0098] Alternatively, non-GMO approaches are using antigens fused to a protein-domains know to anchor itself to the surface of suitable non-pathogenic microorganisms. However, these applications of these approaches may be limited due to concerns already mentioned above. Also, many pathogenic, cancer or allergy antigens may be carbohydrates. The ability of the mentioned approaches to present carbohydrate antigen is still completely unknown. Thus, there is a high need for a new technology that allows to equip non-pathogenic bacteria with pathogenic, cancer or allergy antigens, especially carbohydrate antigens without using genetic modifications.

[0099] Embodiments:

[00100] In some aspects, the present invention relates to a modified microorganism comprising: a cell and a heterologous lipid and/or protein carrier (e.g., said heterologous lipid and/or protein carrier is not expressed or synthetized by said microorganism, e.g. comprising a LysM-based protein carrier as described in Ganesh et al., 2014, Schmidt et al. 2011), said lipid and/or protein carrier comprising: a) a lipid portion, wherein said lipid portion is at least partially associated with an exterior surface of said cell of said modified 32microorganism, wherein said lipid portion comprising: a ceramide-like glycolipid moiety | ;101096 (and/or a ceramide moiety and/or a sphingolipid moiety and/or sphingosine moiety) and/or a fatty acid moiety; preferably said exterior surface of said cell comprising: a cell wall and/or a cell membrane and/or an outer cell membrane and/or a polysaccharide (e.g., capsule polysaccharide); further preferably said lipid portion is at least partially incorporated and/or adhered and/or bound to said exterior surface of said cell; and b) a non-lipid portion, wherein said microorganism is capable of locating and/or displaying said non-lipid portion or fragment thereof onto the exterior surface of said cell, preferably said non-lipid portion comprising a carbohydrate moiety and/or lipopeptide moiety (e.g., a glycosylated lipopeptide moiety, e.g., a carbohydrate antigen); further preferably said non-lipid portion is capable of binding and/or reducing toxicity and/or neutralizing a toxin; optionally, said modified microorganism (e.g., said lipid carrier) further comprising a heterologous steroid moiety, preferably said steroid moiety is cholesterol, a derivative or analogue thereof.

[00101] In some aspects of the present invention, a microorganism may already comprise an endogenous lipid carried (e.g., identical or non-identical with the heterologous lipid carrier of the present invention as decribed herein, e.g., a glycolipid, etc.), wherein: (a) the concentration of said endogenous lipid carrier is increased with said heterologues lipid carrier (e.g., via methods and/or uses as described herein, e.g., loading protocol/s as described, e.g., in the examples section, e.g., Table 1); or (b) said heterologues and endogenous lipid carries are non-identical and are both comprised in the modified microorganism of the present invention (e.g., a microorganism may already comprise an endogenous GM1 saccharide moiety on its surface (e.g., not necessary a whole GM1 molecule as described herein) and is then modified by the means of adding a heterologous lipid carrier (e.g., GM1 molecule).

[00102] In some aspects, the present invention relates to the modified microorganism of the present invention, wherein said non-lipid portion is capable of one or more of the following characteristics: binding and/or reducing toxicity and/or neutralizing a toxin; binding and/or reducing the pathogenicity and/or neutralizing a pathogenic microorganism; binding a receptor of a toxin; and binding a receptor of a pathogenic microorganism.

[00103] In some aspects, the present invention relates to the modified microorganism of the present invention wherein said modified microorganism has one or more of the following characteristics: said cell of said microorganism comprising a cell membrane (e.g. separating the cytoplasm of said cell from the exterior of the cell); preferably said cell membrane further comprising a peptidoglycan; said cell of said microorganism comprising a cell wall, preferably said cell wall comprising a peptidoglycan; said cell of said microorganism comprising a cell membrane, wherein said cell membrane further comprising a peptidoglycan; said cell of said microorganism is not comprising an outer membrane, (e.g., 33separating the prokaryotic periplasm from the exterior of said cell of said microorganism); | 101096 preferably said microorganism is a gram-positive bacterium; said cell of said microorganism comprising an outer membrane (e.g., separating the prokaryotic periplasm from the exterior of said cell of said microorganism); preferably said microorganism is a gram-negative bacterium; said cell of said microorganism comprising a cell wall, preferably said cell wall comprises mannoproteins (e.g., yeast), said cell of said microorganism is not comprising a recombinant and/or fusion polypeptide, preferably said recombinant and/or fusion polypeptide is obtained by the means of artificial genetic manipulation; said microorganism is non-pathogenic.

[00104] In some aspects, the present invention relates to the modified microorganism of the present invention, wherein said lipid carrier is selected from the group consisting of: Monosialotetrahexosylganglioside (GM1), Globotriaosyiceramide (Gb3), Ganglioside GD1a, Gangliosides GM2, GD2, GD1b, GT1b, GT1c, GQ1c, GA1, GM1b, Gangliosides GM3, GD3 und GT3, Gangliosides Gb4, Blood Group Type |, Type 2, Blood Group A, Blood Group B, Blood Group H, Blood Group H Type 1, Blood Group H Type 2, Blood Group H Type 3, Lewis y, Lewis a, Lewis b, Lewis x, H, Sialyl Lewis x, Sialyl Lewis a, Sialyl Lewis b, Sialyl Lewis x, alpha Gal epitope, Gal a1-3Gal31-4GlacNAc, and derivatives and analogues thereof.

[00105] In some aspects, the present invention relates to the modified microorganism of the present invention, wherein said microorganism is naturally-occurring (e.g., non- genetically modified, e.g., not obtained by means of artificial genetic manipulation), preferably said naturally-occurring microorganism is obtainable from one or more of the following sources: microflora of a vertebral organism (e.g., mammalian, avian (e.g., poultry), bovine, porcine, ovine, caprine, leporine, or piscine organism), preferably microflora of a digestive or urinary system (e.g., microflora of a gastro-intestinal tract (GIT) or an urinary tract) or skin microflora of said vertebral organism or microflora of the respiratory tract; further preferably said naturally-occurring microorganism is obtainable from feces or urine or sputum of said vertebral organism; most preferably said mammalian organism is human; and soil microbiota; and microbiota of water such as sea or fresh water; and microbiota of plants or other natural source/s or environment/s.

[00106] In some aspects, the present invention relates to the modified microorganism of the present invention, wherein said microorganism is selected from the group consisting of: a bacterium (e.g., gram-positive or gram-negative (e.g., from genus Bacteroides, e.g., Bacteroides vulgatus) bacterium; preferably said bacterium is a gram-positive bacterium; further preferably said gram-positive bacterium is selected from the genera consisting of: Lactobacillus (e.g., Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus salivarius, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus or Lactobacillus vaginalis), 34

Bifidobacterium (e.g., Bifidobacterium breves Bifidobacterium adolescentis, Bifidobacterium LU101096 longum, Bifidobacterium infantis or Bifidobacterium pseudocatenulatum), Clostridium (e.g., Clostridium perfringens, Clostridium coccides), Enterococcus (e.g., Enterococcus avium, Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinarum, Enterococcus casseliflavus or Enterococcus sp.), Pediococcus (e.g. Pediococcus pentosaceus) and Streptococcus (e.g., Streptococcus salivarius or Streptococcus vestibularis); most preferably said gram-positive bacterium is selected from the group consisting of: Lactobacillus paracasei, Lactobacillus reuteri; and a fungus; preferably said fungus is selected from the group consisting of. Candida yeasts, Saccharomyces yeasts and yeasts in the family Dipodascaceae; further preferably said Dipodascaceae yeasts are Galactomyces, Geotrichum or Saprochaete yeasts.

[00107] In some aspects, the present invention relates to the modified microorganism of the present invention, wherein said microorganism is non-pathogenic, preferably said non- pathogenic is not associated with a mammalian pathological or infectious condition or disease, further preferably said mammalian pathological or infectious condition or disease is a human pathological or infectious condition or disease.

[00108] In some aspects, the present invention relates to the modified microorganism of the present invention, wherein said microorganism is non-toxic to a mammalian host, preferably said microorganism is non-toxic to a human host.

[00109] In some aspects, the present invention relates to the modified microorganism of the present invention, wherein said microorganism is capable of one or more of the following characteristics: binding and/or reducing toxicity and/or neutralizing a toxin; binding and/or reducing the pathogenicity and/or neutralizing a pathogenic microorganism; binding a receptor of a toxin; binding a receptor of a pathogenic microorganism.

[00110] In some aspects, the present invention relates to a composition comprising one or more of the modified microorganisms of the present invention; preferably said composition comprising a mixture of same (e.g., identical, same species, same strain) or different (e.g., non-identical, different species, different strains) modified microorganisms.

[00111] In some aspects, the present invention relates to the composition of the present invention, wherein said composition is a pharmaceutical, diagnostic, probiotic or prebiotic composition.

[00112] In some aspects, the present invention relates to the composition of the present invention, wherein said composition further comprises a pharmaceutical carrier.

[00113] In some aspects, the present invention relates to the composition of the present invention, wherein said composition is suitable for oral or enteral administration.

35

[00114] In some aspects, the present invention relates to a vaccine or adjuvant LU101096 comprising the microorganism of the present invention; preferably said vaccine or adjuvant is suitable for oral or enteral administration.

[00115] The invention is also characterized by the following items:

1. A modified microorganism (e.g., isolated) comprising: i) a cell and ii) a heterologous lipid and/or protein carrier (e.g., said heterologous lipid or protein carrier is not expressed or synthetized by said microorganism), said lipid or protein carrier comprising: a) a lipid portion, wherein said lipid portion is at least partially associated with an exterior surface of said cell of said modified microorganism, wherein said lipid portion comprising: a ceramide-like glycolipid moiety (and/or a ceramide moiety and/or a sphingolipid moiety and/or sphingosine moiety) and/or a fatty acid moiety; preferably said exterior surface of said cell comprising: a cell wall and/or a cell membrane and/or an outer cell membrane and/or a polysaccharide (e.g., capsule polysaccharide), further preferably said lipid portion is at least partially incorporated and/or adhered and/or bound to said exterior surface of said cell; and b) a non-lipid portion, wherein said microorganism is capable of locating and/or displaying said non-lipid portion or fragment thereof onto the exterior surface of said cell, preferably said non-lipid portion comprising a carbohydrate moiety and/or lipopeptide moiety (e.g., a glycosylated lipopeptide moiety, e.g., a carbohydrate antigen); further preferably said non-lipid portion is capable of binding and/or reducing toxicity and/or neutralizing a toxin; iii) optionally, said modified microorganism (e.g., said lipid and/or protein carrier) further comprising a heterologous steroid moiety; preferably said steroid moiety is cholesterol, a derivative or analogue thereof.

2. The modified microorganism according to any one of preceding items, wherein said non-lipid portion is capable of one or more of the following characteristics: i) binding and/or reducing toxicity of and/or neutralizing a toxin; 36ii) binding and/or reducing the pathogenicity and/or neutralizing a pathogenic LU101096 microorganism (e.g., a bacterium or a virus); iii) binding a receptor of a toxin; and iv) binding a receptor of a pathogenic microorganism.

3. The modified microorganism according to any one of preceding items, wherein said modified microorganism has one or more of the following characteristics: i) said cell of said microorganism comprising a cell membrane (e.g., separating the cytoplasm of said cell from the exterior of the cell); preferably said cell membrane further comprising a peptidoglycan; ii) said cell of said microorganism comprising a cell wall, preferably said cell wall comprising a peptidoglycan; iii) said cell of said microorganism comprising a cell membrane (e.g., yeast), wherein said cell membrane further comprising a peptidoglycan; preferably said cell wall comprises a mannportein/s; iv) said cell of said microorganism is not comprising an outer membrane (e.g., separating the periplasm from the exterior of said cell of said microorganism), preferably said microorganism is a gram-positive bacterium.

Vv) said cell of said microorganism comprising an outer membrane (e.g., separating the periplasm from the exterior of said cell of said microorganism); preferably said microorganism is a gram-negative bacterium; vi) said cell of said microorganism is not comprising a recombinant and/or fusion polypeptide, preferably said recombinant and/or fusion polypeptide is obtained by the means of artificial genetic manipulation; vii) said microorganism is non-pathogenic.

4 The modified microorganism according to any one of preceding items, wherein said lipid or protein carrier has one or more of the following characteristics: i) comprising a glyceride moiety; preferably said glyceride moiety comprises at least one fatty acid, further preferably said lipid portion of said lipid and/or protein carrier comprises said glyceride moiety; ii) comprising a ceramide moiety, wherein preferably: 37a) said ceramide moiety is composed of (e.g., comprises) an amino alcohol (e.g., LU101096 sphingosine) and/or a fatty acid, further preferably said lipid portion of said lipid and/or protein carrier comprises said ceramide moiety; and/orb) in said ceramide moiety the first sugar bound to said ceramide is e.g., aglucose, a galactose, a glucose, a mannose, a xylose, a neuraminic acid, a N- acetyl glucosamine, N-acetyl galactosamine or a galacturonic acid;

iii) comprising a carbohydrate moiety; preferably said non-lipid portion of said lipid and/or protein carrier comprises said carbohydrate moiety; further preferably said carbohydrate moiety is B- or a- linked to said ceramide moiety (e.g., as in item

(ii)), further preferably said carbohydrate is not a monosaccharide or a disaccharide moiety; further preferably said carbohydrate is selected from the group consisting of: an oligosaccharide and a polysaccharide, most preferablythe first sugar of the said carbohydrate is a galactose, a glucose, a mannose, axylose, a neuraminic acid, a N-acetyl glucosamine, N-acetyl galactosamine or agalacturonic acid;

iv) comprising one or more polypeptides (e.g., a heterologous recombinant or fusion polypeptide, e.g., a glycosylated polypeptide or an immunologically active polypeptide); preferably said non-lipid portion comprising said one or more polypeptides; further preferably said one or more polypeptides is an enzyme, acytokine or a chemokine, a peptidomimetic compound, an antigen, an antibody

(e.g., a single chain or a single domain antibody), a fragment or derivative thereof;

v) comprising a pharmaceutically active compound; preferably said non-lipid portion comprising said pharmaceutically active compound;

vi) is not comprising a recombinant and/or fusion polypeptide (e.g., an endogenousrecombinant and/or fusion polypeptide), preferably said recombinant and/or fusion polypeptide is obtained by the means of artificial genetic manipulation;

vii) is not expressed or synthetized by said microorganism;

viii)is at least partially expressed or synthetized by said microorganism (e.g. endogenously synthetized (e.g., a microorganism to be modified according to thepresent invention may already comprise an endogenous lipid carried (e.g. identical or non-identical with the heterologous lipid carrier of the present invention, e.g., a glycolipid, etc.), wherein: (a) the concentration of said endogenous lipid carrier is increased with said heterologues lipid carrier (e.g., via methods and/or uses as described herein, e.g., loading protocol/s as described

38herein, e.g., in the examples section, e.g., Table 1); or (b) said heterologues and LU101096 endogenous lipid carries are non-identical, e.g., are both present in the modified microorganism of the present invention (e.g., a microorganism (e.g., bacterium) may already comprise an endogenous GM1 saccharide moiety on the surface (e.g., not necessary the whole GM1 molecule) and is loaded with a heterologous GM1 molecule), preferably said microorganism is capable of expressing or synthetizing a ceramide or sphingolipid moiety (e.g., a Sphingomonas sp. Gram- negative bacterium); ix) said lipid portion is not comprising polypeptides; x) is not comprising a transmembrane polypeptide or a polypeptide membrane anchor domain; xi) is not susceptible to proteolysis (e.g., under physiological conditions, e.g., a temperature in the range of about 20 - 40°C and/or atmospheric pressure of about 1 and/or pH of about 6-8 and/or glucose concentration of about 1-20 mM and/ or atmospheric oxygen concentration); xii) is not immunogenic to a mammalian host, preferably said mammalian host is human; xiii) is immunogenic to a mammalian host, preferably said mammalian host is human: xiv)is not covalently bound to said cell membrane of said cell of said modified microorganism; xv) comprising a glycolipid; xvi)comprising a lipopeptide (e.g., a glycosylated lipopeptide).

5. The modified microorganism according to any one of preceding items, wherein said lipid and/or protein carrier is selected from the group consisting of: i) Monosialotetrahexosylganglioside (GM1), ii) Globotriaosylceramide (Gb3), iii) Ganglioside GD1a, iv) Gangliosides GM2, GD2, GD1b, GT1b, GT1c, GQ1c, GA1, GM1b, v) Gangliosides GM3, GD3 und GT3, vi) Gangliosides Gb4, Blood Group Type |, Type 2, Blood Group A, Blood Group B, Blood Group H, Blood Group H Type 1, Blood Group H Type 2, Blood Group H 39

Type 3, Lewis y, Lewis a, Lewis b, Lewis x, H, Sialyl Lewis x, Sialyl Lewis a, Sialyl LU101096 Lewis b, Sialyl Lewis x, alpha Gal epitope, Gal a1-3GalB1-4GlacNAc, Gal(a 1- 4)Gal(R1-4)GIcNAc-R, Gal(a 1-4)Gal(R1-4)Gic NAc-(R1-3)Gal(31-4)Glc-R, Gal(a 1-4)Gal(R1-4)GIcNAc(B1-2) Man-R vii) any one of the following: 2-6 Sialyl i- | Neu5Ac(a 2-6)Gal(R1-4)GIcNAc(R1-3)Gal(R1-4)[Fuc(a 1-3)]GIcNAc(B1- Lewis x )-R 3'-Sulfo Lewis a HSO3(-3)Gal(R1-3)[Fuc(a1-4)]GIcNAc(B1-)-R 3'-Sulfo Lewis x HSO3(-3)Gal(R1-4)[Fuc(a1-3)]GIcNAc(B1-)-R 6,6'- Disulfo Sialyl Lewis | Neu5Ac(a 2-3)[HSO3(-6)]Gal(R1-4)[Fuc(a 1-3)][HSO3(-6)JGIcNAc(B1-)-

X R 6-Sulfo Lewis x Gal(R1-4)[Fuc(a 1-3)][HSO3(-6)]GlcNAc(R1-)-R 6'-Sulfo Sialyl Lewis x Neu5Ac(a 2-3)[HSO3(-6)]Gal(1-4)[Fuc(a 1-3)]GIcNAc(R1-)-R

STE SLe* Neu5Aca2-3GalB1-4(Fuca1-3)(6-O-Su)GIcNAcB-R 6'-Sia-6- | Su-LacNAc | NeubAca2-6GalB1-4(6-O-Su)GIcNAcB-R | 6-Su- 3° SiaLe“ Neu5Aca2-3GalB1-3(6-O-Su)GIcNAcB-R 6-Su- 3'SLN Neu5Aca2-3GalB1-4(6-O-Su)GIcNAcB-R 3'SLN(Gc) | Neu5Gca2-3GalB1-4GIcNAcB-R 6 SLN(Gc) | Neu5Gca2-6GalB1-4GIcNAcB-R 40

GIcNAcB3’ ß GIcNAcB1-3GalB1-4GIcNAcB-R 0101096

LacNAc

Isomaltotrio

Glca1-6Glca1-6GIcfB-Rse

Chitotriose | GilcNAcB1-4GIcNAB1-4GIcNAcB-Ralpha Galepitope, Gal(a 1-3)Gal(R1-4)GIcNAc-R GIcNAcR1,3ManB81,4GleB - R GalNAca1-3(Fuca1-2)GalB-R Gala1-3(Fuca1-2)GalB-R

Gal+3,4-

GIcNAc GalB1-4(GalB1-3)GIcNAcB-Rasialo- DGalp(R1-3)DGalpNAc(B1-4)DGalp(R1-4)DGicp(B1-1)-Ra - a c(R1- a - Cc -1)- GM1,GA1 P P P P asialo- DGalpNAc(31-4)DGalp(31-4)DGlcp(R1-1)-R a c(R1- a - Cc -1)-

GM2,GA2 P P P

Biood

Group À

Trisaccharide GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-)-R

Blood

Group À

Type 1 GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-3)GIcNAc(B1-)-R Blood

Group À

Type 1

(difucosyl) | GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-3)[Fuc(a 1-4)]GIcNAc(B1-)-R Blood

Group A

Type 2 GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-4)GIcNAc(B1-)-R

41

Blood LU101096 Group A

Type 2

(difucosyl) | GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-4)[Fuc(a 1-3)IGIcNAc(R1-)-R Blood

Group A

Type 3 GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-3)GalNAc(a 1-)-R Blood

Group A

Type 4 GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-3)GalNAc(B1-)-R Blood

Gal(a 1-3)[Fuc (a 1-2)]Gal(R1-3)GIcNAc(B1-3)Gal-R,

Group B

Blood

Group B

Type 2 Gal(a 1-3)[Fuc(a 1-2)]Gal(81-4)GIcNAc(B1-)-R Blood

Group H

Type 1, Fuc(a 1-2)Gal(B1-3)GIcNAc(B1-)-R

Blood

Group H

Type 2, Fuc(a 1-2)Gal(R1-4)GIcNAc(R1-)-R

Blood

Group H

Type 3, Fuc(a 1-2)Gal(R1-3)GalNAc(a 1-)-R

Blood

Group H, Fuc(a 1-2)Gal(B1-)-R Sia6'H

Neu5Acaz2-6(Fuca1-2)GalB1-4GIcNAcB-R

(type 2) 6-LacNAc- TF GalB1-4GIcNAcB1-6(GalB1-3)GalNAca-R

C-Series

Ganglio- R-Gal(R1-3)GalNAc(B1-4)[Neu5Ac(a 2-8)Neu5Ac(a 2-8)NeuSAc(a 2-

42sides 3)]Gal(R1-4)GIc(B1-1)-R LU101096 Oligo- saccharide / Ganglio- tetraosyl Core Structure C-Series Ganglio- sides Oligo- saccharide / Hemato- or Ganglio- Type NeubAc(a 2-8)Neu5Ac(a 2-8)Neu5Ac(a 2-3)Gal-R Cyclic Sialyl 6- Sulfo Lewis x cyclicNeubAc(a 2-3)Gal(b1-4)[Fuc(a 1-3)][HSO3(-6)]GIcNAc-R Dimeric Lewis x Gal(R1-4)[Fuc(a 1-3)]GIcNAc(B1-3)Gal(b1-4)[Fuc(a 1-3)]JGIcNAc(B1-)-R Disialyl Lewis a NeuSAc(a 2-3)Gal(R1-3)[NeuSAc(a 2-6)][Fuc(a 1-4)]GIcNAc(B1-)-R Disialyl Lewis c NeubAc(a 2-3)Gal(R1-3)[Neu5Ac(a 2-6)]GIcNAc(R1-)-R F1 Alpha Gal(b1-4)GIcNAc(b1-6)GalNAc(a 1-)Ser/Thr fucosyl GM1 (Fucal-2Gal81-3GalNAcK1-4[NeuAca2-3]-GalB1-4GIcB1-I-R GA1: (Gg4Cer) | GalB1,3GalNAcB1,4Gal81,4GIcB1-R GA2: (Gg3Cer) | GalNAcB1,4GalB1,4GIicB1-R 43

Gal a1- LU101096 3GalB1- Gal a 1-3GalB1-4GlacNAc - R 4GlacNAc Galili, Gala- Gala1-3GalB1-4GIcNAcB-R 3'LacNAc Gal a 1,4Galß - R er GalNAc- GD1a: (IV3Neu5A cli3NeubAc Gg5Cer) GalNAcB1,4GalB1,3GalNAcB1, 4(Neu5Aca2,3)GalB1, 4GIcB1-R GalR1,3GalNAcR1,4Gal81,4Glcf - R GbOse3Ce r: (Gb3Cer) | Gala1,4GalB1,4GIcB1-R GbOse4Ce r GalNAcB1,3Gala1,4GalB1,4GIcB1-R GD1a: (IV3Neu5A clI3Neu5Ac Gg4Cer) NeuSAca2,3GalB1,3GalNAcB1, 4(Neu5Aca2,3)Gal81,4GIcB1-R GD1b: (I13(Neu5A c)2Gg4Cer) | GalB1,3GalNAcB1, 4(Neu5Aca2,8Neu5Aca2,3)GalB1, 4GIcB1-R GD1b- lactone: II3[NeuSAc-(2-8,1—9)-Neu5Ac]Gg4-R GD1c: (IV3(Neu5A c)2Gg4Cer)| Neu5Aca2,8Neu5Aca2,3GalB1, 3GalNAcB1,4GalB1,4GIcB1-R Neu5Ac a2,3GalB1, 3(Neu5Aca2,6)GalNAcB1, 4Gal81,4GIcB1-R 44

(IV3NeusA LU101096 clli6NeubAcGg4Cer)

GD2:

(I13(Neu5Ac)2Gg3Cer) | GalNAcB1, 4(Neu5Aca2,8Neu5Aca2,3)Gal81, 4GIcB1-R GD3:

(I13(Neu5Ac)2LacCer) | Neu5Aca2,8Neu5Aca2,3GalB1, 4GIcB1-R Neu5Ac(a 2-3)DGalp(R1-1)-Roe

GalNAcR1,3Gala1,4GalB1,4GIcf -R

Globo-H Fuc a 2Gal83GalNAcfK3Gal a 4GalR4GIcBR1-R Gal( R1- 3)GalNAc( B 1-3)Gal( a 1- 4)Gal( B 1- 4)Glc-R GalB3GalNAcB3Gala4GalB4GIc81-Rmonosialyl-

Gb5 SAa3GalB3GalNAcB3Gala4GalB4GIcB1-R disialyl-Gb5 | SAa3GalB3GalNAcB3(SAa2-3)Gala4GalB4GIcB1-R Gala3GalB4GIcB1-R GalNAcB3Gala3GalB4GIcB1-R GalNAca3GalNAcf3Gala4GalB4GIcB1-R

GM1a:r

(II3Neu5Ac

Gg4Cer) | GalB1,3GalNAcB1, 4(Neu5Aca2,3)Gal81, 4GIcB1-R GM1b:

(IV3Neu5AcGg4Cer) | Neu5Aca2,3GalB1,3GalNAcB1, 4GalB1,4GIcB1-R GM3: G 1,4(Neu5Aca2,3)GalB1, 4GIcp1-R

INACB1, 5 , , - (I13NeusAG | CANACRT,4(NeuSAca2,3)Galp1, 4GIcß 45

Gg3Cer) LU101096 GM2b Neu5Ac(a 2-8)Neu5Ac(a 2-3)DGalp(R1-4)DGicp(R1-1 )-R GM3:

(II3Neu5Ac

LacCer) NeuSAca2,3GalB1,4GIcB1-R

GM4:

(I3Neu5AcaGalCer) | Neu5Aca2-3GalB1-R

GP1c:

(IV3(Neu5Ac)21[3(Neu5

Ac)3Gg4Ce | Neu5Aca2,8Neu5Aca2,3GalB1, 3GalNAcB1,4(Neu5Aca2,8Neu5Aca2, r) 8Neu5Aca2,3)GalB1,4GicB1-R

GP1ca:

(IV3Neu5AcllI6Neu5A

C,

I13(Neu5Ac | Neu5Ac a2,3GalB1, 3(Neu5Aca2,6)GaiNAcB1, )3Gg4Cer) |4(Neu5Aca2,8Neu5Aca2, 8Neu5Aca2,3)GalB1,4GIcB1-R GQ1b:

(IV3(Neu5Ac)2113(Neu5

Ac)2Gg4Ce | Neu5Aca2,8Neu5Aca2,3GalB1, 3GalNAcB1,

r) 4(Neu5Aca2,8Neu5Aca2,3)GalB1, 4GicB1-R

GQ1ba:

(IV3(Neu5Ac)21ll6(Neu 5Ac)2Gg4C | Neu5Ac a2,3GalB1, 3(Neu5Aca2,6)GalNAcB1, er) 4(Neu5Aca2,8Neu5Aca2,3)GalB1, 4GIcB1-R GQ1c:

(IV3Neu5Acll3(Neu5A | Neu5Aca2,3GalB1,3GalNAcB1, 4(Neu5Aca2,8Neu5Aca2, c)3Gg4Cer) | 8Neu5Aca2,3)GalB1, 4GIcB1-R

46

GT1 GTIb NeuSAc(a 2-3)DGalp(R1-3)DGalNAc(R1-4)[Neu5Ac(a 2-8)Neu5Ac(a 2- | LU101096 3)]DGalp(R1-4)DGIcp(B1-1)-R GT1a: N(V3(Neu5 Ac)21[3Neu SAcGg4Cer | Neu5Aca2,8Neu5Aca2,3GalB1, 3GalNAcB1,4(Neu5Aca2,3)GalB1, ) AGIcB1-R GT1b: (IV3Neu5A cli3(Neu5A | NeuSAca2,3GalB1,3GalNAcB1, 4(Neu5Aca2,8Neu5Aca2,3) c)2Gg4Cer) | GalB1,4GIcB1-R GT1c: (H3(NeuSA | GalB1,3GalNAcB1, 4(Neu5Aca2,8Neu5Aca2, 8Neu5Aca2,3)GalB1, c)3Gg4Cer)| 4GIcB1-R GT1a: (IV3Neu5A cill6(Neu5 Ac)2Gg4Ce | Neu5Ac a2,3GalB1, 3(Neu5Aca2,6)GalNAcB1, r) 4(Neu5Aca2,3)GalB1,4GicB1-R GalNAcB1,4(Neu5Aca2,8Neu5Aca2, 8Neu5Aca2,3)GalB1, 4GIcB1-R GT3: (I13(NeuAc) 3LacCer) | Neu5Aca2,8Neu5Aca2,8Neu5Aca2, 3GalB1,4GIcB1-R Internal Gal(b1-4)GIcNAc(b1-3)Gal(b1-4)[Fuc(a1-3)]GIcNAc(b1-3)Gal(b1- Lewis x 4)GIcNAc(b1-3)Gal(b1-4)GIcNAc(b1-3)Gal(b1-4)GIc(b1-1)-R Isoglobo GaiNAcB1,3Gala1,3GalB1,4Glcf - R GalB1,4GIcB1-R GalB1,3GIcNAcB1,3Gal81,4GlcB - R Gal(R1-3)[Fuc(a 1-4)]GIcNAc(R1-)-R Fuc(a 1-2)Gal(R1-3)[Fuc(a 1-4)]GIcNAc(R1-)-R Gal(B1-4)[Fuc(a 1-3)]GIcNAc(B1-)-R 47

Lewis y, Fuc(a 1-2)Gal(R1-4)[Fuc(a 1-3)]GIcNAc(B1-)-R LU101096 Fuc a 1,4GIcNAcR1,2Man a 1,3Manß 1,4GIcR-R GalR1,3GalB1,4GalR1,4GIcB - R

N-Acetyl

GD3 NeuSAc(a 2-8)Neu5Ac(a 2-3)Gal(b1-4)Glc(b1-1)-R Gal 1,6GalR 1,6Galß -R GalB1,4GIcNAcB1,3GalB1,4GIcB- R Neu5Ac(a2 -3)Gal(b1) |Neu5Ac(a 2-3)Gal(R1-)-R Neu5Ac(a2 -3)Gal(b1- 3)GalNAc | Neu5Ac(a 2-3)Gal(R1-3)GalNAc-R NeubAc(a2 8)Neu5Ac( a2-3)Gal NeuSAc(a 2-8)Neu5Ac(a 2-3)Gal-R Neu5Aca2-8Neu5Aca2-8Neu5Aca-R Neu5Aca2-3GalB1-4GIcB-R N-Glycolyl GM3 Neu5Gc(a 2-3)Gal(R1-4)GIc(B1-1)-R NOR1 Gal(a 1— 4)GalNAc( BR 1-3)Gal(a a 1- 4)Gal( B 1- 4)Glc-R

Gal(a 1— 4)GalNAc- ( B 1-3)Gal(a 1- 4)GalNAc( R 1-3)Gal(a 1— 4)Gal( NOR2 ß 1- 4)Glc - R GalNAc( ß 1-3)Gal(a 1—4)GalNAc(R1-3)Gal- (a 1-4)Gal( B 1— 4)GIc -R Fuc (a 1-2)Gal(R1-3)GIcNAc(R1-3)Gal-R, NeubAc(a 2-3)DGalp(31-3)DGalNAc(31-4)XNeu5Ac9Ac(a 2- OAc-GT1b 8)NeubSAc(a 2-3)]DGalp(B-4)DGicp(81-1)-R P antigen (Gb4) Gal(a 1- 4)Gal- ( B 1- 4)GicNAc( R 1-3)Gal( B 1- 4)Glc -R 48

Pk Antigen LU101096 (Gb3) Gal(a 1-4)Gal(R1-4)GIc-R A Gala1-4GalB1-4GIcNAcB-R GalNAcR1,4GIcR-R Sialyl Lewis a, Neu5Ac(a 2-3)Gal(R1-3)[Fuc(a 1-4)]GIcNAc(B1-)-R Sialyl Lewis c Neu5Ac(a 2-3)Gal(R1-3)GIcNAc(B1-)-R Sialyl Lewis | NeuSAc(a 2-3)Gal(R1-4)[Fuc(a 1-3)]GIcNAc(B1-3)Gal(81-4)GicNAc(B1- X, 3)Gal(B1-)-R Sialyl Lewis X, NeuSAc(a 2-3)Gal(R1-4)[Fuc(a 1-3)]GIcNAc(B1-)-R Sialyl Lewis | Neu5Ac(a 2-3)Gal(81-4)[Fuc(a 1-3)JGIcNAc(B1-3)Gal(R1-4)GIcNAc(B1- X-i 3)Gal(R1-)-R GalB1-3GIcNAcB1-3GalB1-4GIcB-R GalB1-4GIcNAcB1-3GalB1-4GIcB-R Sialyl-TF NeuSAc(a 2-6) Gal (B1-3) a GalNAc-R, sialyl-Tn Neu5Ac(a 2-6)GalNAc-R, NeuAc-Gal0-3GicNAcB-3GalB-4Glc - R GalR1,4GicR1,3GalR-R Sulfatide: | Sulfate3GalB1-R TF / Core-1 | a Gal (B1-3)aGalNAc-R, 3'-sialyl-TF | Neu5Aca2-3GalB1-3GalNAca-R 6-SiaBTF | Neu5AcB2-6(GalB1-3)GalNAca-R 3-LacNAc- GalB1-4GIcNAB1-3GalNAca-R 49

6-LacNAc- LU101096 T GalB1-4GIcNAB1-6GalNAca-R 6'SLN NeuSAca2-6GalB1-4GIcNAcB-R GIcNAcB1-6(GalB1-3)GalNAca-R GIcNAcB1-3(GIcNAcB1-6)GalNAca-R Trifucosyl- Lewis b Fuc(a 1-2)Gal(b1-3)[Fuc(a 1-4)]GIcNAc(b1-3)Gal(b1-3)[Fuc(a 1- Antigen 4)]GIcNAc(b1-)-R Trifucosyl- Lewis y Fuc(a 1-2)Gal(b1-4)[Fuc(a 1-3)]JGIcNAc(b1-3)Gal(b1-4)[Fuc(a 1- Antigen 3)]GIcNAc(b1-)-R Type 1 (GalNAc(a 1-3)[Fuc(a 1-2)]Gal(B1-3)GIcNAc[Fuc(a 1-4)]-R), Type 1 A GalNAc(a1-3)[Fuc(a 1-2)]Gal(R1 -3)GIcNAc(B1-3)Gal-R, Type 2A | GalNAc(a1-3)[Fuc(a 1-2)]Gal(R1-4)GIcNAC-R, Type 3 A GalNAc(a1-3)[Fuc(a 1-2)]Gal(81-3)GalNAc(R1-3)Gal[Fuca 1-2)-R, GalNAc(a1-3)[Fuc(a 1-2)]Gal(81-3)GalNAc(B1-3)Gal(a 1-4)Gal(B 1- Type 4 A 4)Glc--R, NeuSAc(a 2-3)Gal(31-4)GIcNAc(B1-3)Gal(R1-4)[Fuc(a -3)]GIcNAc(B1-)- VIM-2 R GalNAc(a1-3)[Fuc(a 1-2)]Gal(R1-3)GalNAc(R1-3)Gal(a 1-4)Gal(fB 1- Type 4 A 4)Glc--R, Mana1-3(Mana1-6)Mana-? 3'SLN Neu5Aca2-3GalB1-4GIcNAcB-R Neu5Aca2-6GalB1-4GIcB-R wherein R is one or more of the following: a carbohydrate/s, a peptide/s, a lipid/s, a linker/s and a chemical compound/s or substance/s or molecule/s; or R comprises one or more of the following: a carbohydrate/s, a peptide/s, a lipid/s, a linker/s or a chemical compound/s or substance/s or molecule/s; 50viii)any one of lipid carriers of i)-vii) further coupled to TF disaccharide, Core-1 LU101096 | structure, Tn monosaccharide, Sialyl-TF mono- or disialylated, Sialyl-Tn, Polysialic acid, or mannose-6-phosphate moiety; ix) any one of lipid carriers of i)-viii) further coupled to N-Glycan or O-glycan moiety (e.g., as can be found on glycoproteins or glycopeptides); x) any one of lipid carriers of i)-ix) further coupled to a carbohydrate moiety; Xi) a truncated or elongated derivative of any one of lipid carriers of i)-x); xii) a phosphorylated, sulfated or acetylated derivative of any one of lipid carriers of i) xi); xiii) derivatives and analogues of any of (i)-(xii).

6. The modified microorganism according to any one of preceding items, wherein said microorganism is naturally-occurring, preferably said naturally-occurring microorganism is obtainable from one or more of the following sources: i) microflora or microbiota of an animal, preferably a vertebral organism, preferably microbiota or microflora of a digestive or urogenital system or skin microflora of said vertebral organism; further preferably said naturally- occurring microorganism is obtainable from the gut, feces, oral or nasal cavity, vagina, lung, sputum, other mucus sources or urine of said vertebral organism; most preferably said mammalian organism is human; and ii) soil microflora or microbiota; iii) microbiota from plants, preferably leaves, fruits or berries, or marine ecosystems.

7. The modified microorganism according to any one of preceding items, wherein said microorganism is a gram-positive bacterium, a gram-negative bacterium (e.g., a non- pathogenic gram-negative bacterium), fungus or protozoa.

8. The modified microorganism according to any one of preceding items, wherein said microorganism is non-pathogenic, preferably said non-pathogenic is not associated with a vertebral (e.g., mammalian) pathological or infectious condition or disease, further preferably said vertebral (e.g., mammalian) pathological or infectious condition or disease is a human pathological or infectious condition or disease. 51

9. The modified microorganism according to any one of preceding items, wherein said LU101096 microorganism is non-toxic to a mammalian host, preferably said microorganism is non-toxic to a human host.

10. The modified microorganism according to any one of preceding items, wherein said microorganism is one or more of the following: i) a bacterium; preferably said bacterium is gram-positive or gram-negative bacterium; non-pathogenic and/or opportunistic pathogen; further preferably said bacterium is a gram-positive bacterium; further further preferably said gram-positive bacterium is selected from the genera consisting of: Lactobacillus, Bifidobacterium, Clostridium, Enterococcus, Pediococcus and Streptococcus; most preferably said gram-positive bacterium is selected from the group consisting of: Lactobacillus paracasei, Lactobacillus reuteri: and ii) an opportunistic pathogen selected from the group consisting of genera: Abiotrophia Acanthopleuribacter Acaricomes Acetanaerobacterium Acetatifactor Acetitomaculum Acetivibrio Acetoanaerobium Acetobacter Acetobacterium | Acetofilamentum Acetogenium Thermoanaerobacter Acetohalobium Acetomicrobium Acetonema Acetothermus Acholeplasma Achromatium Achromobacter Acidaminobacter Acidaminococcus Acidianus Acidicaldus Acidicapsa Acidiferrobacter Acidilobus Acidimicrobium Acidiphilium Acidiplasma Acidisoma Acidisphaera Aciditerrimonas Acidithiobacillus Acidobacterium Acidocella Acidomonas Acidothermus Acidovorax Acinetobacter Acrocarpospora Actibacter Actibacterium Actinaurispora Plantactinospora Actinoallomurus Actinoalloteichus Actinobacillus Actinobaculum Actinobispora — synonym: Pseudonocardia Actinocatenispora Actinocorallia Actinokineospora Actinomadura Actinomyces Actinomycetospora Actinophytocola ~~ Actinoplanes ~~ Actinopolymorpha Actinopolyspora Actinopycnidium — synonym: Streptomyces Actinospica Actinosporangium Streptomyces Actinosynnema Actinotalea Adhaeribacter Adlercreutzia Advenella Aegyptianella Aequorivita Aeribacillus Aeriscardovia Aerococcus ~~ Aeromicrobium Aeromonas Aeropyrum Aestuariibacter Aestuariibaculum Aestuariicola Lutimonas Aestuariihabitans Aestuariimicrobium Aestuariispira Afifella Afipia Agaricicola Agarivorans Aggregatibacter Agitococcus Agreia Agrobacterium Agrococcus Agromonas Bradyrhizobium Agromyces Ahrensia Aidingimonas Akkermansia Albibacter Albidiferax Albidovulum Albimonas Alcaligenes Alcanivorax Algibacter Algicola Algimonas Algiphilus Algisphaera Algoriphagus Aliagarivorans Alicycliphilus 52

Alicyclobacillus Aliicoccus Aliidiomarina Aliifodinibius Aliiglaciecola Aliivibrio LU101096 Alishewanella Alistipes Alkalibacillus ~~ Alkalibacter ~~ Alkalibacterium | Alkalibaculum Alkaliflexus ~~ Alkalilimnicola ~~ Alkalimonas Alkaliphilus Alkalispirillum Alkalitalea Alkanibacter Alkanindiges Allisonella Alloactinosynnema Allobacillus Allobaculum Allocatelliglobosispora Allochromatium Allofustis Alloiococcus Allokutzneria Allomonas Allonocardiopsis Alloprevotella Allorhizobium Rhizobium Alloscardovia Alpinimonas Alsobacter Altererythrobacter Alteribacillus Alterococcus Alteromonas Alysiella Amantichitinum Amaricoccus Ameyamaea Aminiphilus Aminivibrio Aminobacter = Aminobacterium Aminomonas Ammonifex Ammoniphilus Amnibacterium Amoebobacter Amorphosporangium Actinoplanes Amorphus Amphibacillus Amphritea Ampullariella Actinoplanes Amycolata Pseudonocardia Amycolatopsis Amycolicicoccus Anaeroarcus Anaerobacillus Anaerobacter Anaerobacterium Anaerobaculum Anaerobiospirilum Anaerobranca Anaerocella Anaerococcus Anaerofilum Anaerofustis Anaeroglobus Anaerolinea Anaeromusa Anaeromyxobacter Anaerophaga Anaeroplasma Anaerorhabdus Anaerosalibacter Anaerosinus Anaerosphaera Anaerosporobacter Anaerostipes Anaerotruncus Anaerovibrio Anaerovirgula Anaerovorax Anaplasma Ancalochloris Ancalomicrobium Ancylobacter Anderseniella Andreprevotia Aneurinibacillus Angiococcus Angulomicrobium Angustibacter Anoxybacillus Anoxynatronum Antarcticimonas Antarctobacter Aquabacter Aquabacterium Aquamicrobium Aquaspirillum Aquibacter Aquicella Aquifex Aquiflexum Aquihabitans Aquimarina Aquimonas Aquincola Aquipuribacter ~~ Aquisalibacillus Aquisalimonas Aquisphaera Aquitalea Arachnia Propionibacterium Arcanobacterium Archaeoglobus Archangium Arcicella Arcobacter Arcticibacter Ardenticatena Arenibacter Arenicella Arenimonas Arenitalea Arhodomonas Aridibacter Armatimonas Arsenicicoccus Arsenophonus Arthrobacter Asaccharobacter Asaccharospora Asaia Asanoa Asinibacterium Aspromonas Arenimonas Asteroleplasma Asticcacaulis Atopobacter Atopobium Atopococcus Atopostipes Aurantimonas Auraticoccus Aureibacter Aureicoccus Aureimonas Aureispira Aureitalea Aureivirga Aureobacterium Microbacterium Auritidibacter ~~ Austwickia ~~ Avibacterium Azoarcus Azohydromonas Azomonas Azomonotrichon Azomonas Azonexus Azorhizobium Azorhizophilus Azospira Azospirillum Azotobacter Azovibrio Bacillus Bacteriolyticum Bacterionema Corynebacterium Bacteriovorax Bacteroides Bactoderma Balnearium Balneatrix Balneimonas Microvirga 53

Balneola Balneomonas BarnesiellaBarrientosiimonas Bartonella Basfia | 101096 Bauldia Bavariicoccus Bdellovibrio Beggiatoa Beijerinckia Belliella Bellilinea Belnapia Beneckea Bergeriella Bergeyella Bermanella Beutenbergia Bhargavaea Bibersteinia Bifidobacterium Bilophila Biostraticola Bisgaardia | 5 Bizionia Blastobacter Blastocatelia Blastochloris Blastococcus Blastomonas Blastopirellula Blattabacterium Blautia Bogoriella Bordetella Borrelia Bosea Bowmanella Brachybacterium Brachymonas Brachyspira Brackiella Bradyrhizobium Branchiibius Brassicibacter ~~ Brenneria Breoghania Brevibacillus Brevibacterium Brevifollis Brevinema Brevundimonas Brochothrix Brockia Brooklawnia Brucella Brumimicrobium Bryantella Marvinbryantia Bryobacter Bryocella Buchnera Budvicia Bulleidia Burkholderia Buttiauxella ~~ Butyricicoccus Butyricimonas Butyrivibrio ~~ Byssovorax Caedibacter Caenibacterium — synonym: Schlegelella Caenimonas Caenispirillum Caldalkalibacillus Caldanaerobacter Caldanaerobius Caldanaerovirga Calderihabitans Calderobacterium Hydrogenobacter Caldibacillus Caldicellulosiruptor Caldicoprobacter Caldilinea Caldimicrobium Caldimonas Caldisericum Caldisphaera Calditerricola Calditerrivibrio Caldithrix Caldivirga Calidifontibacter Caloramator Caloranaerobacter Caloribacterium Calymmatobacterium Klebsiella Camelimonas Caminibacter Caminicella Campylobacter Candidimonas Canibacter Capnocytophaga Capsularis Prevotella Carbophilus Carboxydibrachium Caldanaerobacter Carboxydocella Carboxydothermus Carboxylicivirga Cardiobacterium Carnimonas Carnobacterium Caryophanon Caseobacter — synonym: Corynebacterium Castellaniella Catalinimonas Catellatospora Catellibacterium Gemmobacter Catellicoccus Catelliglobosispora Catenibacterium Catenococcus Catenovulum Catenulispora Catenuloplanes Catonella Caulobacter Cecembia Cedecea Celeribacter Celerinatantimonas Cellulomonas Cellulophaga Cellulosibacter Cellulosilyticum Cellulosimicrobium Cellvibrio Centipeda Cerasibacillus Cerasicoccus Cesiribacter Cetobacterium Chainia Streptomyces Chelativorans Chelatobacter — synonym: Aminobacter Chelatococcus Chelonobacter Chiayiivirga Chimaereicella Algoriphagus Chitinibacter Chitinilyticum Chitinimonas Chitiniphilus Chitinivorax Chitinophaga Chlamydia Chlamydophila Chlorobaculum Chlorobium Chloroflexus Chloroherpeton Chioronema Chondromyces Christensenella Chromatium Chromatocurvus { Chromobacterium Chromohalobacter Chryseobacterium Chryseoglobus Chryseolinea Chryseomicrobium Chryseomonas — synonym: Pseudomonas 54

Chrysiogenes Chthonomonas Chungangia Ciceribacter Citreicella | 101096

Citreimonas Citricoccus Citrobacter Clavibacter Clevelandina Cloacibacillus Cloacibacterium Clostridiisalibacter Clostridium Cnuella Cobetia Cocleimonas Coenonia Cohaesibacter Cohnella Collimonas Collinsella Colwellia

Comamonas Compostimonas Conchiformibius Conexibacter Conglomeromonas Congregibacter Constrictibacter Coprobacillus Coprococcus Coprothermobacter Coraliomargarita Corallibacter Corallococcus Corallomonas Coriobacterium Corynebacterium Cosenzaea Costertonia Couchioplanes Cowdria Ehrlichia Coxiella Crabtreella —

synonym: Shinella Craurococcus Crenotalea Crenothrix Cribrihabitans ,Crinalium“ Cristispira Croceibacter Croceicoccus Croceitalea Crocinitomix Cronobacter Crossiella Cruoricaptor Cryobacterium Cryomorpha Cryptanaerobacter Cryptobacterium Cryptosporangium Cucumibacter Cupriavidus Curtobacterium Curvibacter Cyclobacterium Cycloclasticus

Cystobacter Cytophaga Dactylosporangium Daeguia Dasania Dechloromonas Dechlorosoma — synonym: Azospira Deefgea Deferribacter Deferrisoma Defluvibacter Aquamicrobium Defluvicoccus Defluviimonas Defluviitalea Defluviitoga Dehalobacter Dehalococcoides Dehalogenimonas Dehalospirillum Sulfurospirilum Deinobacter Deinococcus Deinobacterium

Deinococcus Deleya Delftia Demequina Demetria Dendrosporobacter Denitratisoma Denitrobacterium Denitrovibrio Dermabacter Dermacoccus Dermatophilus Derxia Desemzia Desertibacter Desmospora Desulfacinum Desulfarculus Desulfatibacillum Desulfatiferula Desulfatirhabdium Desulfatitalea Desulfitibacter Desulfitispora Desulfitobacterium Desulfobacca

Desulfobacter Desulfobacterium Desulfobacula Desulfobaculum Desulfobotulus Desulfobulbus Desulfocapsa Desulfocella Desulfococcus Desulfoconvexum Desulfocurvus Desulfofaba Desulfofrigus Desulfofustis Desulfoglaeba Desulfohalobium Desulfoluna Desulfomicrobium Desulfomonas Desulfovibrio Desulfomonile Desulfomusa Desulfofaba Desulfonatronobacter

Desulfonatronospira Desulfonatronovibrio Desulfonatronum Desulfonauticus Desulfonema Desulfonispora Desulfopila Desulforegula Desulforhabdus | Desulforhopalus Desulfosalsimonas Desulfosarcina Desulfosoma Desulfospira Desulfosporosinus Desulfotalea Desulfothermus Desulfotignum Desulfotomaculum Desulfovermiculus Desulfovibrio Desulfovirga

Desulfovirgula Desulfurella Desulfurispira Desulfurispirilum Desulfurispora Desulfurivibrio Desulfurobacterium Desulfurococcus Desulfurolobus Acidianus Desulfuromonas Desulfuromusa Dethiobacter Dethiosulfatibacter

55

Dethiosulfovibrio Devosia Devriesea Dialister _ Diaminobutyricimonas LU101096 Diaphorobacter Dichelobacter Dichotomicrobium Dickeya Dictyoglomus Dietzia Dinoroseobacter Diplocalyx Diplorickettsia Dissulfuribacter Dokdonella Dokdonia Dolosigranulum Domibacillus Donghaeana Nonlabens Donghicola Dongia Dorea Draconibacterium Duganella Dyadobacter Dyella Dysgonomonas Echinicola Echinimonas Ectothiorhodosinus Ectothiorhodospira Edaphobacter Edwardsiella Effluviibacter Pontibacter Eggerthella Paraeggerthella Eggerthella Eggerthia Ehrlichia Eikenella Eilatimonas Eionea Eisenbergiella Ekhidna Elioraea Elizabethkingia Elstera Elusimicrobium Elytrosporangium Streptomyces Empedobacter Emticicia Endobacter Endozoicomonas Enhydrobacter Enhygromyxa Ensifer Enteractinococcus Enterobacter Enterococcus Enterorhabdus Enterovibrio Entomoplasma Eoetvoesia Eperythrozoon Epibacterium Epilithonimonas Eremococcus Erwinia Erysipelothrix Erythrobacter Erythromicrobium Erythromonas Blastomonas Escherichia Ethanoligenens Eubacterium Eudoraea Euzebya Euzebyella Ewingella Excellospora Actinomadura Exiguobacterium Exilispira Extensimonas Fabibacter Facklamia Faecalibacterium Faenia Saccharopolyspora Falcivibrio Falsibacillus Falsiporphyromonas Falsirhodobacter Falsochrobactrum Fangia Ferribacterium Ferrimicrobium Ferrimonas Ferriphaselus Ferrithrix Ferroglobus Ferroplasma Ferrovibrio | Ferruginibacter ~~ Fervidicella Fervidicoccus Fervidicola Fervidobacterium Fibrella Fibrisoma Fibrobacter Fictibacillus Filibacter Filifactor Filimonas Filobacillus Filomicrobium Fimbrimonas Finegoldia Flagelimonas Flammeovirga Flaviflexus Flavihumibacter Flavimonas — synonym: Pseudomonas Flaviramulus Flavisolibacter Flavitalea Flavivirga Flavobacterium Flavonifractor Flectobacillus Flexibacter Flexistipes Flexithrix Flexivirga Flindersiella Fluoribacter Fluviicola Fluviimonas Fodinibacter Fodinibius Fodinicola Fodinicurvata Fontibacillus Fontibacter Fonticella Fontimonas Formivibrio Formosa Francisella Frankia Frateuria Fretibacter Fretibacterium Friedmanniella Frigoribacterium Frischella Frondicola Frondihabitans Frondihabitans Fructobacillus Fuchsiella Fulvibacter Fulvimarina Fulvimonas Fulvitalea Fulvivirga Fundibacter Alcanivorax Fusibacter Fusicatenibacter Fusobacterium Gaetbulibacter Gaetbulicola Gaetbulimicrobium Aquimarina Gaiella Galbibacter Galbitalea Galenea Gallaecimonas Gallibacterium Gallicola Gallionella Gangjinia Garciella Gardnerella Gelidibacter Gelria Gemella Geminicoccus Gemmata Gemmatimonas Gemmiger Gemmobacter 56

Geoalkalibacter Geobacillus Geobacter Geodermatophilus Geofilum | 101096 Geoglobus Geojedonia Geomicrobium Geopsychrobacter Georgenia Georgfuchsia Geosporobacter Geothermobacter Geothrix Geotoga Geovibrio Gibbsiella Giesbergeria Gilliamella Gillisia Gilvibacter Gilvimarinus Glaciecola Glaciibacter Glaciihabitans Glaciimonas Globicatella Gluconacetobacter Gluconobacter Glycocaulis Glycomyces Goodfellowiella ~~ Gordonia Gordonibacter Gracilibacillus ~~ Gracilibacter Gracilimonas Grahamella Bartonella Gramella Granulibacter Granulicatella Granulicella | Granulicoccus Granulosicoccus Grimontia Gryllotalpicola Guggenheimella Gulbenkiania Gulosibacter Haematobacter Haemobartonella Haemophilus Hafnia Hahella Haladaptatus Halalkalibacillus Halalkalicoccus Halanaerobacter Halanaerobaculum Halanaerobium Halarchaeum Halarsenatibacter Haliangium Haliea Halioglobus Haliscomenobacter Hallella Haloactinobacterium Haloactinopolyspora Haloactinospora Haloarchaeobius Haloarchaeum Haloarcula Halobacillus Halobacterium Halobacteroides Halobaculum Halobellus Halobiforma Halocella Halochromatium Halococcus Haloechinothrix Haloferax Haloferula Halogeometricum Haloglycomyces Halogranum Halohasta Haloincola Halolactibacillus Halolamina Halomethanococcus Halomicroarcula Halomicrobium Halomonas Halonatronum Halonotius Halopelagius Halopenitus Halopiger Haloplanus Haloplasma Haloquadratum Halorhabdus Halorhodospira Halorientalis Halorubellus Halorubrobacterium Halorubrum Halorussus Halosarcina Halogeometricum Halosimplex Halospina Halostagnicola Halotalea Haloterrigena Halothermothrix ~~ Halothiobacillus Halovenus Halovibrio Halovivax Hamadaea Hansschlegelia Haploangium Hasllibacter Hazenella Helcobacillus ~~ Helcococcus Helicobacter Heliimonas Heliobacillus Heliobacterium Heliophilum Heliorestis Heliothrix Hellea Henriciella Hephaestia Herbaspirillum Herbiconiux Herbidospora Herminiimonas Herpetosiphon Hespellia Hippea Hirschia Histophilus Hoeflea Holdemania Hollandina Holophaga Holospora Homoserinimonas Hongia Hongiella Hoppeia Howardella Hoyosella Huaishuia Huanghella Humibacillus Humibacter Humicoccus Nakamurella Humihabitans Intrasporangium Humitalea Hungatella Hwangdonia Hwanghaeicola Hyalangium Hydrocarboniphaga Hydrogenibacillus Hydrogenimonas Hydrogenispora Hydrogenivirga Hydrogenoanaerobacterium Hydrogenobacter Hydrogenobaculum Hydrogenophaga Hydrogenophilus Hydrogenothermus | Hydrogenovibrio Hydrotalea Hylemonella Hymenobacter Hyperthermus 57

Hyphomicrobium Hyphomonas Hyunsoonleella lamia Ideonella Idiomarina | y101096 Ignatzschineria Ignavibacterium Ignavigranum Ignicoccus Ignisphaera llumatobacter Hyobacter Imperialibacter Imtechella Indibacter Inhella Inquilinus Insolitispirilum Intestinibacter Intestinimonas Intrasporangium lodobacter Isobaculumisochromatium Isoptericola Isosphaera Jahnella Janibacter Jannaschia Janthinobacterium Jatrophihabitans Jejudonia Jejuia Jeongeupia Jeotgalibaca Jeotgalibacillus Jeotgalicoccus Jhaorihella Jiangella Jishengella Johnsonella Jonesia Jonquetella Joostella Kaistella Chryseobacterium Kaistia Kallotenue Kandleria Kangiella Kerstersia Ketogulonicigenium Kibdelosporangium Kiloniella Kineococcus Kineosphaera Kineosporia Kingella Kinneretia Kistimonas Kitasatoa Streptomyces Kitasatospora Klebsiella Klugiella Kluyvera Knoellia Kocuria Kofleria Komagataeibacter Kordia Kordiimonas Koreibacter - synonym: Paraoerskovia Kosakonia Koserella Kosmotoga Kozakia Krasilnikovia Kribbella Kribbia Kriegella Krokinobacter Dokdonia Kroppenstedtia Ktedonobacter [Ktedobacter] Kurthia Kushneria Kutzneria Kyrpidia Kytococcus Labedaea Labedella Labrenzia Labrys Laceyella Lachnoanaerobaculum Lachnobacterium Lachnospira Lacibacter Lacibacterium Lacinutrix Lacticigenium Lactivibrio Lactobacillus Lactococcus Lactonifactor Lactosphaera Trichococcus Lactovum Lamprobacter Lamprocystis Lampropedia Lapillicoccus Laribacter Larkinella Lautropia Lawsonia Leadbetterella Lebetimonas Lechevalieria Leclercia Leeia Leeuwenhoekiella Legionella Leifsonia Leisingera Lelliottia Leminorella Lentibacillus Lentibacter Lentilitoribacter Lentisphaera Lentzea Leptobacterium Leptolinea Leptonema Leptospira Leptospirilum Leptothrix Leptotrichia Leucobacter Leuconostoc Leucothrix Levilinea Levinea Lewinella ,Liberibacter" Lihuaxuella Limibacter Limimonas Limnobacter Limnohabitans Lishizhenia Listeria Listonella Litoreibacter Litoribacillus Litoribacter Litoricola Litorilinea Litorimicrobium Litorimonas Litorisediminicola Loktanella Lonepinella Longilinea Longimycelium Longispora Lonsdalea Lucibacterium Vibrio Luedemannella Luminiphilus | Lutaonella Luteibacter Luteibaculum Luteimicrobium Luteimonas Luteipulveratus Luteivirga Luteococcus Luteolibacter Lutibacter Lutibaculum Lutimaribacter Lutimonas Lutispora Lysinibacillus Lysinimicrobium Lysinimonas Lysobacter Lyticum Macellibacteroides Macrococcus Macromonas Magnetococcus Magnetospira Magnetospirillum Magnetovibrio Mahella Malikia Malonomonas Mameliella Mangrovibacter Mangrovibacterium Mangroviflexus Mangrovimonas 58

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Mannheimia Maribacter Maribaculum Henriciella Maribius Maricaulis LU101096

Marichromatium Maricurvus Marihabitans Marinactinospora Marinibacillus

Jeotgalibacillus Marinicauda Marinicella Marinicola Roseivirga Marinifilum

Mariniflexile Marinilabilia Marinilactibacillus Mariniluteicoccus

Marinimicrobium | Marininema Mariniradius ~~ Marinithermus Marinitoga

Marinivirga Algibacter Marinobacter Marinobacterium Marinococcus

Marinomonas Marinoscillum Marinospirillum Marinovum Mariprofundus

Marisediminicola Marispirillum Maritalea Maritimibacter Maritimimonas

Marivirga Marivita Marixanthomonas Marmoricola Martelella Marvinbryantia

Massilia Mechercharimyces Megamonas Meganema Megasphaera

Meiothermus Melaminivora Melghirimyces Melioribacter Melissococcus

Melitea — synonym: Spongiibacter Melittangium Meniscus Meridianimaribacter

Mesoaciditoga Mesoflavibacter Mesonia Mesophilobacter Mesoplasma

Mesorhizobium Mesotoga Metallibacterium Metallosphaera Metascardovia

Alloscardovia Methanimicrococcus Methanobacterium Methanobrevibacter

Methanocalculus Methanocaldococcus Methanocella Methanococcoides

Methanococcus Methanocorpusculum Methanoculleus Methanofollis

Methanogenium Methanohalobium Methanohalophilus Methanolacinia

Methanolinea Methanolobus Methanomassiliicoccus Methanomethylovorans

Methanomicrobium Methanoplanus Methanopyrus Methanoregula

Methanosaeta Methanosalsum Methanosarcina Methanosphaera

Methanospirillum Methanothermobacter Methanothermococcus

Methanothermus Methanothrix Methanosaeta Methanotorris Methermicoccus

Methylarcula Methylibium Methylobacillus Methylobacter Methylobacterium

Methylocaldum Methylocapsa Methyloceanibacter Methylocella

Methylococcus Methylocystis Methyloferula Methylogaea Methylohalobius

Methylohalomonas Methyloligella | Methylomarinovum Methylomarinum

Methylomicrobium Methylomonas Methylonatrum Methyloparacoccus

Methylophaga Methylophilus Methylopila Methylorhabdus Methylorosula

Methylosarcina Methylosinus Methylosoma Methylosphaera Methylotenera

Methylothermus Methyloversatilis Methylovirgula Methylovorus Methylovulum

Micavibrio Microaerobacter Microbacterium Microbispora Microbulbifer

Microcella Micrococcus Microcycius Ancylobacter Microellobosporia

Microlunatus Micromonas Micromonospora Micropolyspora Micropruina

Microscilla Microsphaera Nakamurella Microterricola Microtetraspora

Microvirga Microvirgula Millisia Miniimonas Mitsuaria Mitsuokella

Mobilicoccus Mobiluncus Modestobacter Modicisalibacter Moellerella

59

Mogibacterium Moheibacter Mongoliicoccus Mongoliitalea Mooreia Moorella | J101096 Moraxella Morganella Moritella Morococcus Moryella Motilibacter Mucilaginibacter MucispirilumMumia Murdochiella Muricauda Muricoccus Roseomonas Muriicola Murinocardiopsis Myceligenerans Mycetocola Mycobacterium Mycoplana Mycoplasma Myroides Myxococcus Naasia Nafulsella Nakamurella Namhaeicola Nannocystis Natranaerobaculum Natranaerobius Natranaerovirga Natrialba Natribacillus = Natrinema Natroniella Natronincola Natronoarchaeum Natronobacillus Natronobacterium Natronocella Natronococcus Natronoflexus Natronolimnobius Natronomonas Natronorubrum Natronovirga Naumannella Nautella Nautilia Naxibacter Massilia Necropsobacter Negativicoccus Neiella Neisseria Neoasaia Neochlamydia Neokomagataea Neorickettsia Neptuniibacter Neptunomonas Nereida Nesiotobacter Nesterenkonia Nevskia Nguyenibacter Niabella Niastella Nibrella Nibribacter Nicoletella Nisaea Nitratifractor Nitratireductor Nitratiruptor Nitriliruptor Nitrincola Nitritalea Nitrobacter Nitrococcus Nitrolancea Nitrosococcus Nitrosolobus Nitrosospira Nitrosomonas Nitrosospira Nitrospina Nitrospira Nocardia Nocardioides Nocardiopsis Nonlabens Nonomuraea Noviherbaspirillum Novispirillum Novosphingobium Nubsella Obesumbacterium Oceanibacterium Oceanibaculum Oceanibulbus Oceanicaulis Oceanicella Oceanicola Oceanimonas Oceanirhabdus Oceaniserpentilia Oceanisphaera Oceanitalea Oceanithermus Oceanobacillus Oceanobacter Oceanococcus Oceanospirilum Oceanotoga Ochrobactrum Octadecabacter Odoribacter Oenococcus Oerskovia Ohtaekwangia Okibacterium Oleibacter Oleiphilus Oleispira Oligella Oligosphaera Oligotropha Olivibacter Olleya Olsenella Opitutus Orbus Orenia Oribacterium Oribaculum Porphyromonas Orientia Ornatilinea Ornithinibacillus Ornithinibacter ~~ Ornithinicoccus ~~ Ornithinimicrobium ~~ Ornithobacterium Oryzihumus Oscillibacter Oscillochloris ~~ Oscillospira = Otariodibacter Ottowia Owenweeksia Oxalicibacterium Oxalobacter Oxalophagus Oxobacter Pacificibacter Paenalcaligenes Paenibacillus Paenirhodobacter Paenisporosarcina Paenochrobactrum Palaeococcus Palleronia Paludibacter Paludibacterium Panacagrimonas Pandoraea Pannonibacter Pantoea Papillibacter Parabacteroides Parachlamydia Paracoccus Paracraurococcus Paraeggerthella Paraferrimonas Paraherbaspirilum Paralactobacillus Lactobacillus Paralcaligenes Paraliobacillus Paramoritella Paraoerskovia Parapedobacter ~~ Paraperlucidibaca Paraprevotella Parapusillimonas Pararhodobacter Pararhodospirilum Parascardovia Parasegetibacter 60

Parasphingopyxis Parasporobacterium Parasutterella LU101096 ParvibacterParvibaculum Parvimonas Parvularcula Pasteurella Pasteuria Patulibacter Paucibacter Paucimonas =… Paucisalibacillus Pectinatus Pectobacterium Pediococcus Pedobacter Pedomicrobium Pelagibaca Pelagibacillus Terribacillus Pelagibacterium Pelagibius Pelagicoccus Pelagicola Pelagimonas Pelczaria Pelistega Pelobacter ,Pelodictyon® Pelomonas Pelosinus Pelospora Pelotomaculum Peptococcus Peptoniphilus Peptostreptococcus Peredibacter Perexilibacter Periucidibaca Persephonella Persicirhabdus Persicitalea Persicivirga Persicobacter Petrimonas Petrobacter Tepidiphilus Petrolinea Petrotoga Pfennigia Lamprocystis Phaeobacter Phaeochromatium Phaeocystidibacter Phaeospirillum Phaeovibrio Phascolarctobacterium Phaselicystis Phaseolibacter Phenylobacterium Phocaeicola Phocoenobacter Phorcysia Photobacterium Photorhabdus | Phreatobacter Phycicoccus Phycicola Phycisphaera Phyllobacterium Phytohabitans Phytomonospora Pibocella Picrophilus Pigmentiphaga Pilibacter Pilimelia Pillotina Pimelobacter Pirella Pirellula Piscibacillus Piscicoccus Pisciglobus Piscinibacter Piscirickettsia Planctomyces Planifilum Planktomarina Planktotalea Planobacterium Chryseobacterium Planobispora Planococcus Planomicrobium Planomonospora Planopolyspora Catenuloplanes Planosporangium Planotetraspora Plantactinospora Plantibacter Plasticicumulans Pleionea Pleomorphobacterium Pleomorphomonas Plesiocystis Plesiomonas Pluralibacter Polaribacter Polaromonas Polyangium Polycladomyces Polymorphobacter Polymorphospora Polynucleobacter Pontibaca Pontibacillus Pontibacter Ponticaulis Ponticoccus Pontimonas Pontirhabdus Algibacter Porphyrobacter Porphyromonas Porticoccus Poseidonocella Postechiella Pragia Prauserella Prevotella Pricia Primorskyibacter Prochloron Prochlorothrix Profundibacterium Prolinoborus Prolixibacter Promicromonospora Propionibacter Propionivibrio Propionibacterium Propionicicella Propioniciclava Propionicimonas Propioniferax Propionigenium Propionimicrobium Propionispira Propionispora Propionivibrio Prosthecobacter Prosthecochloris | Prosthecomicrobium Proteiniborus Proteiniclasticum Proteiniphilum Proteinivorax Proteocatella Proteus Protomonas Methylobacterium Providencia Pseudacidovorax Pseudahrensia Pseudaminobacter Pseudarcicella Pseudenhygromyxa Pseudidiomarina Idiomarina Pseudoalteromonas Pseudoamycolata Pseudonocardia Pseudobacteroides Pseudobutyrivibrio Pseudocaedibacter 61

Pseudochrobactrum Pseudoclavibacter Pseudoduganella LU101096

| Pseudoflavonifractor Pseudofulvibacter Pseudofulvimonas Pseudogulbenkiania Pseudohaliea Pseudokineococcus Pseudolabrys

Pseudomaricurvus Pseudomonas Pseudonocardia Pseudopedobacter

Pseudoramibacter Pseudorhodobacter Pseudorhodoferax Pseudoruegeria Pseudosphingobacterium Pseudospirillum Pseudosporangium Pseudoteredinibacter Pseudothermotoga Pseudovibrio Pseudoxanthobacter Pseudoxanthomonas Pseudozobellia Psychrilyobacter Psychrobacillus

Psychrobacter Psychroflexus Psychroglaciecola Psychromonas

Psychroserpens Psychrosphaera Pullulanibacillus Puniceibacterium Puniceicoccus Pusillimonas Pustulibacterium Pyramidobacter Pyrinomonas

Pyrobaculum Pyrococcus Pyrodictium Pyrolobus Pyxidicoccus [Pyxicoccus]

Quadricoccus = Quatrionicoccus Quadrisphaera Quatrionicoccus Quinella

Rahnella Ralstonia Ramlibacter Raoultella Rapidithrix Rarobacter

Rathayibacter =~ Rehaibacterium Reichenbachia Reichenbachiella Reichenbachiella Reinekea Renibacterium Reyranella Rhabdochromatium

Rhabdothermus Rheinheimera Rhizobacter Rhizobium Rhizomicrobium

Rhizomonas Sphingomonas Rhizorhabdus Rhizorhapis Rhodanobacter

Rhodobaca Rhodobacter Rhodobium Rhodoblastus Rhodocista Rhodococcus

Rhodocyclus Rhodocytophaga Rhodoferax Rhodoglobus Rhodoligotrophos Rhodomicrobium Rhodonellum Rhodopila Rhodopirellula Rhodoplanes Rhodopseudomonas Rhodospira Rhodospirillum Rhodothalassium

Rhodothermus Rhodovarius Rhodovibrio Rhodovulum Rickettsia Rickettsiella

Riemerella Rikenella Rivibacter Rivicola Robertkochia Robiginitalea

Robiginitomaculum Robinsoniella Rochalimaea Bartonella Romboutsia Roseateles Roseburia Roseibaca Roseibacillus Roseibacterium Roseibium

Roseicitreum Roseicyclus Roseiflexus Roseimicrobium Roseinatronobacter

Roseisalinus Roseivirga Roseivivax ~~ Rosenbergiella Roseobacter

Roseococcus Roseomonas Roseospira Roseospirilum Roseovarius Rothia

Ruania Rubellimicrobium Rubribacterium Rubricoccus Rubrimonas Rubritalea Rubritepida Rubrivirga Rubrivivax Rubrobacter Rudaea

Rudaeicoccus Rudaibacter Rudanella Ruegeria Rufibacter Rugamonas

Rugosimonospora Ruminobacter Ruminococcus Rummeliibacillus Runella

Sabulilitoribacter Saccharibacillus Saccharibacter Saccharicrinis

Saccharobacter Saccharococcus Saccharofermentans Saccharomonospora Saccharophagus Saccharopolyspora Saccharospirillum Saccharothrix

Sagittula Salana Salegentibacter Salibacillus Virgibacillus Salicola

62

Salimesophilobacter ~~ Salimicrobium Salinactinospora Salinarchaeum LU101096 Salinarimonas Salinibacillus Salinibacter Salinibacterium Salinicoccus Salinicola Salinigranum Salinihabitans Salinimicrobium Salinimonas Salinirepens Salinirubrum Salinisphaera Salinispora Salinivibrio Salipiger Salirhabdus Salisaeta Salisediminibacterium Saliterribacillus Salmonella15 Salsuginibacillus Samsonia Sandaracinobacter Sandaracinus Sandarakinorhabdus Sandarakinotalea Nonlabens Sanguibacter Saprospira Sarcina Sarcobium Legionella Saxeibacter Nakamurella Scardovia Schineria Ignatzschineria Schlegelella Schleiferia Schlesneria Schumannella Schwartzia Sciscionella Sebaldella Sedimentibacter Sedimenticola Sediminibacillus Sediminibacter Sediminibacterium Sediminicola Sediminihabitans Sediminimonas Sediminitomix Segetibacter Segniliparus Seinonella Sejongia Chryseobacterium Selenihalanaerobacter Seleniivibrio Selenomonas Seliberia Senegalimassilia Seohaeicola Seonamhaeicola Serinibacter Serinicoccus Serpens Serpula Brachyspira Serpulina Brachyspira Serratia Sharpea Shewanella Shigella Shimazuella Shimia Shimwellia Shinella Shivajiella Shuttleworthia Siansivirga Silanimonas Silicibacter Ruegeria Silvimonas Simiduia Simkania Simonsiella Simplicispira Singularimonas Solimonas Singulisphaera Sinibacillus Sinobaca Sinobacter Solimonas Sinobacterium Sinococcus Sinobaca Sinomicrobium Sinomonas Sinorhizobium Sinosporangium Siphonobacter Skermanella Skermania Slackia Smaragdicoccus Smithella Sneathia Sneathiella Snodgrassella Snuella Sodalis Soehngenia Solibacillus Solimonas Solirubrobacter Solitalea Solobacterium Soonwooa Sorangium Spelaeicoccus Sphaerisporangium Sphaerobacter Sphaerochaeta Sphaerosporangium Sphaerotilus Sphingobacterium Sphingobium Sphingomicrobium Sphingomonas Sphingopyxis Sphingorhabdus Sphingosinicella Spinactinospora Spirilliplanes Spirillospora Spirillum Spirochaeta Spiroplasma Spirosoma Spongiibacter Spongiibacterium Spongiimonas Spongiispira Sporacetigenium Sporanaerobacter Sporichthya Sporobacter Sporobacterium Sporocytophaga Sporohalobacter Sporolactobacillus Sporolituus Sporomusa Sporosalibacterium Sporosarcina Sporotalea Pelosinus Sporotomaculum Stackebrandtia Stakelama Staleya Sulfitobacter Stanierella Aquimarina Staphylococcus Staphylothermus Stappia Starkeya Stella Stenothermobacter Nonlabens Stenotrophomonas Stenoxybacter Steroidobacter Sterolibacterum Stetteria Stibiobacter Stigmatella Stomatobaculum Stomatococcus Rothia Streptacidiphilus 63

Streptoalloteichus Streptobacillus Streptococcus Streptohalobacillus LU101096 Streptomonospora Streptomyces Streptosporangium Streptoverticillium Streptomyces Stygiolobus Subdoligranulum Subsaxibacter Subsaximicrobium Subtercola Succinatimonas Succiniclasticum Succinimonas Succinispira Succinivibrio Sulfitobacter Sulfobacillus Sulfolobus Sulfophobococcus Sulfuricella Sulfuricurvum Sulfurihydrogenibium Sulfurimonas Sulfurisoma Sulfurisphaera Sulfuritalea Sulfurivirga Sulfurococcus Sulfurospirillum Sulfurovum Sungkyunkwania Sunxiuginia Sutterella Suttonella Swaminathania Swingsia Symbiobacterium Symbiotes Synergistes Syntrophaceticus | Syntrophobacter Syntrophobotulus Syntrophococcus Syntrophomonas Syntrophorhabdus Syntrophospora Syntrophomonas Syntrophothermus Syntrophus Tabrizicola Tahibacter Taibaiella Tamlana Tamlicoccus Tannerella Tanticharoenia Taonella Tardiphaga Tateyamaria Tatlockia Tatumella Taylorella Tectibacter Teichococcus Roseomonas Telluria Telmatobacter Telmatocola Telmatospirillum Tenacibaculum Tenuibacillus Tepidamorphus Tepidanaerobacter Tepidibacillus Tepidibacter Tepidicella Tepidimicrobium Tepidimonas Tepidiphilus Terasakiella Teredinibacter Terrabacter Terracoccus Terribacillus Terriglobus Terrimicrobium Terrimonas Terrisporobacter Tersicoccus Tessaracoccus Tetragenococcus Tetrasphaera Tetrathiobacter Advenella Texcoconibacillus Thalassobacillus Thalassobacter Thalassobaculum Thalassobius Thalassococcus Thalassolituus Thalassomonas Thalassospira Thalassotalea Thauera Thermacetogenium Thermaerobacter Thermanaeromonas Thermanaerovibrio Thermasporomyces Thermicanus Thermincola Thermithiobacillus Thermoactinomyces Thermoactinospora Thermoanaerobacter Thermoanaerobacterium Thermoanaerobaculum Thermoanaerobium Thermobacillus Thermobacteroides Thermobifida Thermobispora Thermobrachium Thermocatellispora Thermochromatium Thermocladium Thermococcoides Kosmotoga Thermococcus Thermocrinis Thermocrispum Thermodesulfatator Thermodesulfobacterium Thermodesulfobium Thermodesulforhabdus Thermodesulfovibrio Thermodiscus Thermofilum Thermoflavifilum Thermoflavimicrobium Thermoflexus Thermogemmatispora Thermogymnomonas Thermohalobacter Thermohydrogenium Thermoleophilum Thermolithobacter Thermolongibacillus Thermomarinilinea Thermomicrobium Thermomonas Thermomonospora Thermonema Thermophagus Thermoplasma Thermopolyspora Thermoproteus Thermosediminibacter Thermosinus 64

Thermosipho Thermosphaera Thermosporothrix Thermosulfidibacter | 101096 Thermosulfurimonas Thermosyntropha Thermotalea Thermoterrabacterium Carboxydothermus Thermothrix Thermotoga Thermotunica Thermovenabulum Thermovibrio Thermovirga Thermovorax Thermovum Thermus Thioalbus Thioalkalibacter ~~ Thioalkalicoccus Thioalkalimicrobium Thioalkalispira Thioalkalivibrio Thiobaca Thiobacillus Thiobacter Thiobacterium Thiocapsa Thioclava Thiococcus Thiocystis Thiodictyon Thiofaba Thioflavicoccus Thiohalobacter Thiohalocapsa Thiohalomonas Thiohalophilus Thiohalorhabdus Thiohalospira Thiolamprovum Thiomargarita Thiomicrospira Thiomonas Thiopedia Thiophaeococcus Thioploca Thioprofundum Thioreductor Thiorhodococcus Thiorhodospira Thiorhodovibrio Thiosphaera Paracoccus Thiospira Thiospirilum Thiothrix Thiovirga Thiovulum Thorsellia Tindallia Tissierella Tistlia Tistrella Tolumonas Tomitella Tonsilliphilus Toxothrix Trabulsiella Tranquillimonas Treponema Trichlorobacter Geobacter Trichococcus Tropheryma Tropicibacter Tropicimonas Truepera Trueperella Tsukamurella Tuberibacillus Tumebacillus Turicella Turicibacter Turneriella Uliginosibacterium Ulvibacter Umboniibacter Umezawaea Undibacterium Ureaplasma Ureibacillus Uruburuella Vadicella Vagococcus Vallitalea Vampirovibrio Varibaculum Variovorax Vasilyevaea Veillonella Venenivibrio Verminephrobacter Verrucomicrobium Verrucosispora Vibrio Vibrionimonas Victivallis Virgibacillus Virgisporangium Viridibacillus Vitellibacter Vitreoscilla Vogesella Volcaniela Halomonas Volucribacter Vulcanibacillus Vulcaniibacterum Vulcanisaeta WVulcanithermus Waddlia Wandonia Wangia Zunongwangia Wautersia Cupriavidus Wautersiella Weeksella Weissella Wenxinia Wenyingzhuangia Wigglesworthia Williamsia Winogradskyella Wohifahrtimonas Wolbachia Wolinella Woodsholea Xanthobacter Xanthomonas Xenophilus Xenorhabdus Xiangella Xylanibacter Prevotella Xylanibacterium Xylanimicrobium Xylanimonas Xylella Xylophilus Yangia Yania Yaniella Yaniella Yeosuana Yersinia Yimella Yokenella — synonym: Koserella Yonghaparkia Youngiibacter Yuhushiella Zavarzinella Zavarzinia Zeaxanthinibacter Zhangella Maritalea Zhihengliuella Zhongshania Zhouia Zimmermannella = Pseudoclavibacter Zobellella Zobellia Zoogloea Zooshikella Zunongwangia Zymobacter Zymomonas Zymophilus; iii) said microorganism is non-pathogenic and/or opportunistic pathogen and is selected from the group consisting of genera as defined in item (ii) herein; 65iv) said microorganism is Gram-positive, non-pathogenic and/or opportunistic LU101096 pathogen and is selected from the group consisting of genera as defined in item (ii) herein; v) a fungus; preferably said fungus is selected from the group consisting of: Candida yeasts, Saccharomyces yeasts and yeasts in the family Dipodascaceae; further preferably said Dipodascaceae yeasts are Galactomyces, Geotrichum or Saprochaete yeasts, most preferably said Saccharomyces yeast is Saccharomyces boulardii, S. cerevisiae, S. pastorianus, or Schizosaccharomyces pombe; vi) a protozoa, said protozoa being non-pathogenic to human, preferably said protozoa is Chilomastix mesnili, Endolimax nana, Entamoeba coli, Entamoeba dispar, Entamoeba hartmanni, Entamoeba polecki or lodamoeba buetschiii.

11. The modified microorganism according to any one of preceding items, wherein said microorganism is capable of one or more of the following characteristics: i) binding and/or reducing toxicity of and/or neutralizing a toxin; ii) binding and/or reducing the pathogenicity and/or neutralizing a pathogenic microorganism; iii) binding a receptor of a toxin; iv) binding a receptor of a pathogenic microorganism; v) associating with one of more of the following: glycolipids, lipopeptides, immunological active peptides, proteins (e.g., anti-inflammatory cytokines), antibodies (e.g., single domain antiboidies), antibody fragments and derivatives thereof coupled to said lipid carrier (e.g., its lipid portion); vi) eliciting or modulating an immune response; vii) monitoring development of a disease and/or assessing the efficacy of a therapy of a disease; viii) delivering a pharmaceutically active compound, preferably said delivering is to a mucosal tissue of a subject (e.g., patient); ix) screening a candidate compound for activity against a disease; x) screening for microorganisms capable of binding the lipid carrier according to any one of preceding items.

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12. A composition comprising one or more of the modified microorganisms according to LU101096 any one of preceding items; preferably said composition comprising a mixture of same or different modified microorganisms according to any one of preceding items.

13. The composition according to any one of preceding items, wherein said composition is a pharmaceutical, diagnostic, probiotic or prebiotic composition.

14. The composition according to any one of preceding items, wherein said composition further comprises a pharmaceutical carrier.

15. The composition according to any one of preceding items, wherein said composition is suitable for oral, enteral, dermal, topical, urogenital, inhaltational administration.

16. A vaccine or adjuvant comprising the microorganism or composition according to any one of preceding items (e.g., said microorganism is pathogenic and/or immunogenic, e.g., wherein said microorganism is alive and/or attenuated and/or pasteurized and/or inactivated and/or non-living and/or dead).

17. The vaccine or adjuvant comprising the microorganism or composition according to any one of preceding items, wherein said vaccine or adjuvant is suitable for oral or enteral administration.

18. A method for producing or isolating a modified microorganism comprising: i) a cell and ii) a heterologous lipid and/or protein carrier (e.g., said heterologous lipid and/or protein carrier is not expressed or synthetized by said microorganism), said lipid carrier comprising: a) a lipid portion, wherein said lipid portion is at least partially associated with an exterior surface of said cell of said modified microorganism, wherein said lipid portion comprising: a ceramide-like glycolipid moiety (and/or a ceramide moiety and/or a sphingolipid moiety and/or sphingosine moiety) and/or a fatty acid moiety; preferably said exterior surface of said cell comprising: a cell wall and/or a cell membrane and/or an outer cell membrane and/or a polysaccharide (e.g., capsule polysaccharide); further preferably said lipid portion is at least partially incorporated and/or adhered and/or bound to said exterior surface of said cell; and b) a non-lipid portion, wherein said microorganism is capable of locating and/or displaying said non-lipid portion or fragment thereof onto the 67exterior surface of said cell, preferably said non-lipid portion comprising | J101096 a carbohydrate moiety and/or lipopeptide moiety (e.g., a glycosylated lipopeptide moiety, e.g., a carbohydrate antigen); further preferably said non-lipid portion is capable of binding and/or reducing toxicityand/or neutralizing a toxin;

iii) optionally, said modified microorganism (e.g., said lipid carrier) further comprising a heterologous steroid moiety; preferably said steroid moiety is cholesterol, a derivative or analogue thereof;

said method comprising:

i’) providing:

a’) a microorganism or mixture of microorganisms, preferably said microorganism or mixture of microorganisms are naturally-occurring; further preferably said naturally-occurring is obtainable from one or more of the following sources:

aa’) microflora or microbiota of an animal, preferably a vertebral organism, preferably microbiota or microflora of a digestive or urogenital system or skin microflora of said vertebral organism; further preferably said naturally-occurring microorganism is obtainable from the gut, feces, oral or nasalcavity, vagina, lung, sputum, other mucus sources or urine of said vertebral organism; most preferably said mammalian organism is human; and microflora of a vertebral organism, preferably microflora of a digestive or urinary system or skin microflora of said vertebral organism; further preferably saidnaturally-occurring microorganism is obtainable from feces or urine of said vertebral organism; most preferably said mammalian organism is human; bb’) soil microflora or microbiota; cc’) microbiota of plants, preferably leaves, fruits or berries,

or marine or freshwater ecosystems;

optionally, isolating and/or enriching said microorganism or mixture of microorganisms from said one or more sources; 68i") adding a loading medium to said microorganism or mixture of microorganisms, | ;101096 preferably said loading medium is a culture medium, e.g., BSM (Bifidobacterium specific Medium), SBSM (Self Made Bifidobacterium specific Medium) LB (Lysogeny broth), MRS (de Man, Rogosa and Sharpe Medium) or WC (Wilkins and Chalgren Medium); further preferably said loading medium is a buffer solution, further most preferably said adding is the resuspending of said microorganism or mixture of microorganisms in said loading medium; ii’) adding said solubilized heterologous lipid carrier (e.g., according to any one of preceding items) to the suspension of said microorganism or mixture of microorganisms in said loading medium; preferably said adding is carried out at a temperature in the range between about 4°C and about 70°C, preferably between about 12 and about 65°C, further preferably between about 18°C and about 50°C, further preferable between about 23°C and about 46°C, further preferably in the range from about 30°C to about 55°C; further preferably said temperature is selected from the group consisting of: about 30, 37, 46 and 55°C; iv’) optionally, isolating said microorganism or mixture of microorganisms comprising said lipid carrier, preferably by the means of affinity purification (e.g., with one or more toxins, e.g., CT, STX1/2 and/or LT, pathogen receptor(s) or parts thereof).

19. The method for producing or isolating a modified microorganism according to any one of preceding items further comprising: Vv’) culturing said microorganism in a culture medium under growth conditions and isolating said microorganism from said culture medium; vi’) optionally, discarding the microorganism or mixture of microorganisms not comprising said lipid carrier (e.g., after isolating said microorganism or mixture of microorganisms comprising said lipid carrier, e.g., as in item (iv’) of claim 18); vi’) optionally, identifying said isolated microorganism, preferably by the means of mass spectroscopy and/or nucleic acid sequencing and/or microbiological analysis and/or biochemical analysis; viii') optionally, carring out said method under different condition (preferably with another growth medium and/or loading medium and/or under different loading conditions (e.g., of the lipid carrier to the microorganism) and/or growing conditings (e.g., temperature and/or pH) (e.g., different from those in claim 18)) 69and evaluating the effect of said condition on the association (e.g., strength, LU101096 | stability, concentration, amount of) of said lipid carrier with said microorganism.

20. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein said loading medium comprises cholesterol.

21. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein said culturing is carried out under anaerobic conditions.

22. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein said culture medium is not suitable for an optimal growth of said microorganism, preferably said culture medium is not suitable for supporting a highest growth rate and/or shortest generation time for said microorganism.

23. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein said culture medium comprises propionic acid.

24. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein said growth conditions are not suitable for an optimal growth of said microorganism, preferably said growth conditions are not suitable for supporting a highest growth rate and/or shortest generation time (e.g. about 20 minutes) for said microorganism.

25. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein said growth conditions comprise one or more of the following: i) a temperature in the range between about 4°C and about 70°C, preferably between about 12°C and about 60°C, further preferably between about 18°C and about 50°C, further preferable between about 23°C and about 46°C, further preferably in the range from about 30°C to about 55°C; most preferably said temperature is selected from the group consisting of: about 30, 37, 46 and 55°C.

ii) a pH in the range between about 1 and about 10, preferably between about 3 and about 9, further preferably between about 4 and about 8, further preferably in the range from about 3 to about 7, most preferably pH in the range from about 5 to about 6.

26. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein prior to adding said solubilized heterologous lipid carrier the suspension of said microorganism is having an optical density (OD) in the range between about OD 0.1 and about OD 100, preferably between about OD 0.3 and about 70

| OD 30, preferably between about OD 0.5 and about OD 10, further preferably between | 101096 about OD 1 and about OD5, most preferably in the range from about OD 1 to about OD 2, further most preferably said OD is measured at a wavelength in the range from about 540 to about 660 nm, further most preferably OD is measured at a wavelength of about 600 nm.

27. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein said adding of the solubilized heterologous lipid carrier is carried out at a temperature in the range between about 4°C and about 70°C, preferably between about 12°C and about 60°C, further preferably between about 18°C and about 55°C, further preferably between about 23°C and about 50°C, further preferably in the range from about 30°C to about 46°C; most preferably said temperature is selected from the group consisting of: about 30, 37, 46 and 55°C.

28. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein said microorganism is a gram-positive bacterium, a gram- negative bacterium (e.g., a non-pathogenic gram-negative bacterium) or a fungus.

29. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein said microorganism is non-pathogenic, preferably said non-pathogenic is not associated with a vertebral (e.g., mammalian) pathological or infectious condition or disease, further preferably said vertebral (e.g., mammalian) pathological or infectious condition or disease is a human pathological or infectious condition or disease.

30. The modified microorganism according to any one of preceding items, wherein said microorganism is non-toxic to a mammalian host, preferably said microorganism is non-toxic to a human host.

31. The modified microorganism according to any one of preceding items, wherein said microorganism is one or more of the following: i) a bacterium; preferably said bacterium is gram-positive or gram-negative bacterium; non-pathogenic and/or opportunistic pathogen; further preferably said bacterium is a gram-positive bacterium; further preferably said gram- positive bacterium is selected from the genera consisting of: Lactobacillus, Bifidobacterium, Clostridium, Enterococcus, Pediococcus and Streptococcus; most preferably said gram-positive bacterium is selected from the group consisting of: Lactobacillus paracasei, Lactobacillus reuteri; and 71ii) an opportunistic pathogen selected from the group consisting of genera: | y101096 Abiotrophia Acanthopleuribacter Acaricomes Acetanaerobacterium Acetatifactor Acetitomaculum Acetivibrio Acetoanaerobium Acetobacter Acetobacterium Acetofilamentum Acetogenium Thermoanaerobacter Acetohalobium Acetomicrobium Acetonema Acetothermus Acholeplasma Achromatium Achromobacter Acidaminobacter Acidaminococcus Acidianus Acidicaldus Acidicapsa Acidiferrobacter Acidilobus Acidimicrobium Acidiphilium Acidiplasma Acidisoma Acidisphaera Aciditerrimonas Acidithiobacillus Acidobacterium Acidocella Acidomonas Acidothermus Acidovorax Acinetobacter Acrocarpospora Actibacter Actibacterium Actinaurispora Plantactinospora Actinoallomurus Actinoalloteichus Actinobacillus Actinobaculum Actinobispora — synonym: Pseudonocardia Actinocatenispora Actinocorallia Actinokineospora Actinomadura Actinomyces Actinomycetospora ~~ Actinophytocola Actinoplanes Actinopolymorpha Actinopolyspora Actinopycnidium — synonym: Streptomyces Actinospica Actinosporangium Streptomyces Actinosynnema Actinotalea Adhaeribacter Adlercreutzia Advenella Aegyptianella Aequorivita Aeribacillus Aeriscardovia Aerococcus Aeromicrobium Aeromonas Aeropyrum Aestuariibacter Aestuariibaculum Aestuariicola Lutimonas Aestuariihabitans Aestuariimicrobium Aestuariispira Afifella Afipia Agaricicola Agarivorans Aggregatibacter Agitococcus Agreia Agrobacterium Agrococcus Agromonas Bradyrhizobium Agromyces Ahrensia Aidingimonas Akkermansia Albibacter Albidiferax Albidovulum Albimonas Alcaligenes Alcanivorax Algibacter Algicola Algimonas Algiphilus Algisphaera Algoriphagus Aliagarivorans Alicycliphilus Alicyclobacillus Aliicoccus Aliidiomarina Aliifodinibius Aliiglaciecola Aliivibrio Alishewanella Alistipes Alkalibacillus = Alkalibacter ~~ Alkalibacterium Alkalibaculum Alkaliflexus Alkalilimnicola Alkalimonas Alkaliphilus Alkalispirillum Alkalitalea Alkanibacter Alkanindiges Allisonella Alloactinosynnema Allobacillus Allobaculum Allocatelliglobosispora Allochromatium Allofustis Alloiococcus Allokutzneria Allomonas Allonocardiopsis Alloprevotella Allorhizobium Rhizobium Alloscardovia Alpinimonas Alsobacter Altererythrobacter Alteribacillus Alterococcus Alteromonas Alysiella Amantichitinum Amaricoccus Ameyamaea Aminiphilus Aminivibrio Aminobacter Aminobacterium ~~ Aminomonas Ammonifex Ammoniphilus Amnibacterium Amoebobacter Amorphosporangium Actinoplanes Amorphus Amphibacillus Amphritea Ampullariella Actinoplanes Amycolata Pseudonocardia Amycolatopsis Amycolicicoccus Anaeroarcus 72

Anaerobacillus Anaerobacter Anaerobacterium Anaerobaculum | 101096 | Anaerobiospirilum Anaerobranca Anaerocella Anaerococcus Anaerofilum Anaerofustis Anaeroglobus Anaerolinea Anaeromusa Anaeromyxobacter Anaerophaga Anaeroplasma Anaerorhabdus Anaerosalibacter Anaerosinus Anaerosphaera Anaerosporobacter Anaerostipes Anaerotruncus Anaerovibrio | Anaerovirgula Anaerovorax Anaplasma Ancalochloris Ancalomicrobium Ancylobacter Anderseniella Andreprevotia Aneurinibacillus Angiococcus Angulomicrobium Angustibacter Anoxybacillus Anoxynatronum Antarcticimonas Antarctobacter Aquabacter Aquabacterium Aquamicrobium Aquaspirilum Aquibacter Aquicella Aquifex Aquiflexum Aquihabitans Aquimarina Aquimonas Aquincola Aquipuribacter Aquisalibacillus Aquisalimonas Aquisphaera Aquitalea Arachnia Propionibacterium Arcanobacterium Archaeoglobus Archangium Arcicella Arcobacter Arcticibacter Ardenticatena Arenibacter Arenicella Arenimonas Arenitalea Arhodomonas Aridibacter Armatimonas Arsenicicoccus Arsenophonus Arthrobacter Asaccharobacter Asaccharospora Asaia Asanoa Asinibacterium Aspromonas Arenimonas Asteroleplasma Asticcacaulis Atopobacter Atopobium Atopococcus Atopostipes Aurantimonas Auraticoccus Aureibacter Aureicoccus Aureimonas Aureispira Aureitalea Aureivirga Aureobacterium Microbacterium Auritidibacter ~~ Austwickia ~~ Avibacterium Azoarcus Azohydromonas Azomonas Azomonotrichon Azomonas Azonexus Azorhizobium Azorhizophilus Azospira Azospirillum Azotobacter Azovibrio Bacillus Bacteriolyticum Bacterionema Corynebacterium Bacteriovorax Bacteroides Bactoderma Balnearium Balneatrix Balneimonas Microvirga Balneola Balneomonas BarnesiellaBarrientosiimonas Bartonella Basfia Bauldia Bavariicoccus Bdellovibrio Beggiatoa Beijerinckia Belliella Bellilinea Belnapia Beneckea Bergeriella Bergeyella Bermanella Beutenbergia Bhargavaea Bibersteinia Bifidobacterium Bilophila Biostraticola Bisgaardia Bizionia Blastobacter Blastocatella Blastochloris Blastococcus Blastomonas Blastopirellula Blattabacterium Blautia Bogoriella Bordetella Borrelia Bosea Bowmanella Brachybacterium Brachymonas Brachyspira Brackiella Bradyrhizobium Branchiibius Brassicibacter Brenneria Breoghania Brevibacillus Brevibacterium Brevifollis Brevinema Brevundimonas Brochothrix Brockia Brooklawnia Brucella Brumimicrobium Bryantella Marvinbryantia Bryobacter Bryocella Buchnera Budvicia Bulleidia Burkholderia Buttiauxella Butyricicoccus Butyricimonas Butyrivibrio Byssovorax Caedibacter Caenibacterium — synonym: Schilegelella Caenimonas

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Caenispirillum Caldalkalibacillus Caldanaerobacter Caldanaerobius | 3101096 Caldanaerovirga Calderihabitans Calderobacterium Hydrogenobacter Caldibacillus Caldicellulosiruptor Caldicoprobacter Caldilinea Caldimicrobium Caldimonas Caldisericum Caldisphaera Calditerricola Calditerrivibrio Caldithrix Caldivirga Calidifontibacter Caloramator Caloranaerobacter Caloribacterium Calymmatobacterium Klebsiella Camelimonas Caminibacter Caminicella Campylobacter Candidimonas Canibacter Capnocytophaga Capsularis Prevotella Carbophilus Carboxydibrachium Caldanaerobacter Carboxydocella Carboxydothermus Carboxylicivirga Cardiobacterium Carnimonas Carnobacterium Caryophanon Caseobacter — synonym: Corynebacterium Castellaniella Catalinimonas Catellatospora Catellibacterium Gemmobacter Catellicoccus Catelliglobosispora Catenibacterium Catenococcus Catenovulum Catenulispora Catenuloplanes Catonella Caulobacter Cecembia Cedecea Celeribacter Celerinatantimonas Cellulomonas Cellulophaga Cellulosibacter Cellulosilyticum Cellulosimicrobium Cellvibrio Centipeda Cerasibacillus Cerasicoccus Cesiribacter Cetobacterium Chainia Streptomyces Chelativorans Chelatobacter — synonym: Aminobacter Chelatococcus Chelonobacter Chiayiivirga Chimaereicella Algoriphagus Chitinibacter Chitinilyticum Chitinimonas Chitiniphilus Chitinivorax Chitinophaga Chlamydia Chlamydophila Chiorobaculum Chlorobium Chloroflexus Chloroherpeton Chloronema Chondromyces Christensenella Chromatium Chromatocurvus Chromobacterium Chromohalobacter Chryseobacterium Chryseoglobus Chryseolinea Chryseomicrobium Chryseomonas — synonym: Pseudomonas Chrysiogenes Chthonomonas Chungangia Ciceribacter Citreicella Citreimonas Citricoccus Citrobacter Clavibacter Clevelandina Cloacibacillus Cloacibacterium Clostridiisalibacter Clostridium Cnuella Cobetia Cocleimonas Coenonia Cohaesibacter Cohnella Collimonas Collinsella Colwellia Comamonas Compostimonas Conchiformibius Conexibacter | 30 Conglomeromonas Congregibacter Constrictibacter Coprobacillus Coprococcus Coprothermobacter Coraliomargarita Corallibacter Corallococcus Corallomonas Coriobacterium Corynebacterium Cosenzaea Costertonia Couchioplanes Cowdria Ehrlichia Coxiella Crabtreella — synonym: Shinella Craurococcus Crenotalea Crenothrix Cribrihabitans ,Crinalium“ Cristispira Croceibacter Croceicoccus Croceitalea Crocinitomix Cronobacter Crossiella Cruoricaptor Cryobacterium Cryomorpha Cryptanaerobacter ~~ Cryptobacterium Cryptosporangium Cucumibacter 74

Cupriavidus Curtobacterium Curvibacter Cyclobacterium Cycloclasticus | 3101096 | Cystobacter Cytophaga Dactylosporangium Daeguia Dasania Dechloromonas | Dechlorosoma — synonym: Azospira Deefgea Deferribacter Deferrisoma Defluvibacter Aquamicrobium Defluvicoccus Defluviimonas Defluviitalea Defluviitoga Dehalobacter Dehalococcoides Dehalogenimonas Dehalospirilum Sulfurospirillum Deinobacter Deinococcus Deinobacterium Deinococcus Deleya Delftia Demequina Demetria Dendrosporobacter Denitratisoma Denitrobacterium Denitrovibrio Dermabacter Dermacoccus Dermatophilus Derxia Desemzia Desertibacter Desmospora Desulfacinum Desulfarculus Desulfatibacillum Desulfatiferula Desulfatirhabdium Desulfatitalea Desulfitibacter Desulfitispora Desulfitobacterium Desulfobacca Desulfobacter Desulfobacterium Desulfobacula Desulfobaculum Desulfobotulus Desulfobulbus Desulfocapsa Desulfocella Desulfococcus Desulfoconvexum Desulfocurvus Desulfofaba Desulfofrgus Desulfofustis Desulfoglaeba Desulfohalobium Desulfoluna Desulfomicrobium Desulfomonas Desulfovibrio Desulfomonile Desulfomusa Desulfofaba Desulfonatronobacter Desulfonatronospira Desulfonatronovibrio Desulfonatronum Desulfonauticus Desulfonema Desulfonispora Desulfopila Desulforegula Desulforhabdus Desulforhopalus Desulfosalsimonas Desulfosarcina Desulfosoma Desulfospira Desulfosporosinus Desulfotalea Desulfothermus Desulfotignum Desulfotomaculum Desulfovermiculus Desulfovibrio Desulfovirga Desulfovirgula Desulfurella Desulfurispira Desulfurispirilum Desulfurispora Desulfurivibrio Desulfurobacterium Desulfurococcus Desulfurolobus Acidianus Desulfuromonas Desulfuromusa Dethiobacter Dethiosulfatibacter Dethiosulfovibrio Devosia Devriesea Dialister Diaminobutyricimonas Diaphorobacter Dichelobacter Dichotomicrobium Dickeya Dictyoglomus Dietzia Dinoroseobacter Diplocalyx Diplorickettsia Dissulfuribacter Dokdonella Dokdonia Dolosigranulum Domibacillus Donghaeana Nonlabens Donghicola Dongia Dorea Draconibacterium Duganella Dyadobacter Dyella Dysgonomonas Echinicola Echinimonas Ectothiorhodosinus Ectothiorhodospira Edaphobacter Edwardsiella Effluvibacter Pontibacter Eggerthella Paraeggerthella Eggerthella Eggerthia Ehrlichia Eikenella Eilatimonas Eionea Eisenbergiella Ekhidna Elioraea Elizabethkingia Elstera Elusimicrobium Elytrosporangium Streptomyces Empedobacter Emticicia Endobacter Endozoicomonas Enhydrobacter Enhygromyxa Ensifer Enteractinococcus Enterobacter Enterococcus Enterorhabdus Enterovibrio Entomoplasma Eoetvoesia Eperythrozoon Epibacterium Epilithonimonas 75

Eremococcus Erwinia Erysipelothrix Erythrobacter Erythromicrobium | 101096 Erythromonas Blastomonas Escherichia Ethanoligenens Eubacterium Eudoraea Euzebya Euzebyella Ewingella Excellospora Actinomadura Exiguobacterium Exilispira Extensimonas Fabibacter Facklamia Faecalibacterium Faenia Saccharopolyspora Falcivibrio Falsibacillus Falsiporphyromonas Falsirhodobacter Falsochrobactrum Fangia Ferribacterium Ferrimicrobium Ferrimonas Ferriphaselus Ferrithrix | Ferroglobus Ferroplasma Ferrovibrio Ferruginibacter Fervidicella | Fervidicoccus Fervidicola Fervidobacterium Fibrella Fibrisoma Fibrobacter Fictibacillus Filibacter Filifactor Filimonas Filobacillus Filomicrobium Fimbriimonas Finegoldia Flagellimonas Flammeovirga Flaviflexus Flavihumibacter Flavimonas — synonym: Pseudomonas Flaviramulus Flavisolibacter ~~ Flavitalea Flavivirga Flavobacterium Flavonifractor Flectobacillus Flexibacter Flexistipes Flexithrix Flexivirga Flindersiella Fluoribacter Fluviicola Fluviimonas Fodinibacter Fodinibius Fodinicola Fodinicurvata Fontibacillus Fontibacter Fonticella Fontimonas Formivibrio Formosa Francisella Frankia Frateuria Fretibacter Fretibacterium Friedmanniella Frigoribacterium Frischella Frondicola Frondihabitans Frondihabitans Fructobacillus Fuchsiella Fulvibacter Fulvimarina Fulvimonas Fulvitalea Fulvivirga Fundibacter Alcanivorax Fusibacter Fusicatenibacter Fusobacterium Gaetbulibacter Gaetbulicola Gaetbulimicrobium Aquimarina Gaiella Galbibacter Galbitalea Galenea Gallaecimonas Gallibacterium Gallicola Gallionella Gangjinia Garciella Gardnerella Gelidibacter Gelria Gemella Geminicoccus Gemmata Gemmatimonas Gemmiger Gemmobacter Geoalkalibacter Geobacillus Geobacter Geodermatophilus Geofilum Geoglobus Geojedonia Geomicrobium Geopsychrobacter Georgenia Georgfuchsia Geosporobacter Geothermobacter Geothrix Geotoga Geovibrio Gibbsiella Giesbergeria Gilliamella Gillisia Gilvibacter Gilvimarinus Glaciecola Glaciibacter Glaciihabitans Glaciimonas Globicatella Gluconacetobacter Gluconobacter Glycocaulis Glycomyces Goodfellowiella ~~ Gordonia Gordonibacter Gracilibacillus ~~ Gracilibacter Gracilimonas Grahamella Bartonelia Gramella Granulibacter Granulicatella Granulicella Granulicoccus Granulosicoccus Grimontia Gryllotalpicola Guggenheimella Gulbenkiania Gulosibacter Haematobacter Haemobartonella Haemophilus Hafnia Hahella Haladaptatus Halalkalibacillus Halalkalicoccus Halanaerobacter Halanaerobaculum Halanaerobium Halarchaeum Halarsenatibacter Haliangium Haliea Halioglobus Haliscomenobacter Hallella

76 |

Haloactinobacterium Haloactinopolyspora Haloactinospora Haloarchaeobius | 3101096 Haloarchaeum Haloarcula Halobacillus Halobacterium Halobacteroides Halobaculum Halobellus Halobiforma Halocella Halochromatium Halococcus Haloechinothrix Haloferax Haloferula Halogeometricum Haloglycomyces Halogranum Halohasta Haloincola Halolactibacillus Halolamina Halomethanococcus Halomicroarcula Halomicrobium Halomonas Halonatronum Halonotius Halopelagius Halopenitus Halopiger Haloplanus Haloplasma Haloquadratum Halorhabdus Halorhodospira Halorientalis Halorubellus Halorubrobacterium Halorubrum Halorussus Halosarcina Halogeometricum Halosimplex Halospina Halostagnicola Halotalea Haloterrigena Halothermothrix ~~ Halothiobacillus Halovenus Halovibrio Halovivax Hamadaea Hansschlegelia Haploangium Hasliibacter Hazenella Helcobacillus ~~ Helcococcus Helicobacter Helimonas Heliobacillus Heliobacterium Heliophilum Heliorestis Heliothrix Hellea Henriciella Hephaestia Herbaspirilum Herbiconiux Herbidospora Herminiimonas Herpetosiphon Hespellia Hippea Hirschia Histophilus Hoeflea Holdemania Hollandina Holophaga Holospora Homoserinimonas Hongia Hongiella Hoppeia Howardella Hoyosella Huaishuia Huanghella Humibaciilus Humibacter Humicoccus Nakamurella Humihabitans Intrasporangium Humitalea Hungatella Hwangdonia Hwanghaeicola Hyalangium | Hydrocarboniphaga Hydrogenibacillus Hydrogenimonas Hydrogenispora Hydrogenivirga Hydrogenoanaerobacterium Hydrogenobacter Hydrogenobaculum Hydrogenophaga Hydrogenophilus Hydrogenothermus Hydrogenovibrio Hydrotalea Hylemonella Hymenobacter Hyperthermus Hyphomicrobium Hyphomonas Hyunsoonleella lamia Ideonella Idiomarina Ignatzschineria Ignavibacterium Ignavigranum Ignicoccus Ignisphaera llumatobacter llyobacter Imperialibacter Imtechella Indibacter Inhella Inquilinus Insolitispirilum Intestinibacter Intestinimonas Intrasporangium lodobacter Isobaculumisochromatium Isoptericola Isosphaera Jahnella Janibacter Jannaschia Janthinobacterium Jatrophihabitans Jejudonia Jejuia Jeongeupia Jeotgalibaca Jeotgalibacillus Jeotgalicoccus Jhaorihella Jiangella Jishengella Johnsonella Jonesia Jonquetella Joostella _ Kaistella Chryseobacterium Kaistia Kallotenue Kandleria Kangiella Kerstersia Ketogulonicigenium Kibdelosporangium Kiloniella Kineococcus Kineosphaera Kineosporia Kingella Kinneretia Kistimonas Kitasatoa Streptomyces Kitasatospora Klebsiella Klugiella Kluyvera Knoellia Kocuria Kofleria Komagataeibacter Kordia Kordiimonas Koreibacter - synonym: Paraoerskovia 77

Kosakonia Koserella Kosmotoga Kozakia Krasilnikovia Kribbella Kribbia LU101096 Kriegella Krokinobacter Dokdonia Kroppenstedtia Ktedonobacter [Ktedobacter] Kurthia Kushneria Kutzneria Kyrpidia Kytococcus Labedaea Labedella Labrenzia Labrys Laceyella Lachnoanaerobaculum Lachnobacterium Lachnospira Lacibacter Lacibacterium Lacinutrix Lacticigenium Lactivibrio Lactobacillus Lactococcus Lactonifactor Lactosphaera Trichococcus Lactovum Lamprobacter Lamprocystis Lampropedia Lapillicoccus Laribacter Larkinella Lautropia Lawsonia Leadbetterella Lebetimonas Lechevalieria Leclercia Leeia Leeuwenhoekiella Legionella Leifsonia Leisingera Lelliottia Leminorella Lentibacillus Lentibacter Lentilitoribacter Lentisphaera Lentzea Leptobacterium Leptolinea Leptonema Leptospira Leptospirillum Leptothrix Leptotrichia Leucobacter Leuconostoc Leucothrix Levilinea Levinea Lewinella ,Liberibacter” Lihuaxuella Limibacter Limimonas Limnobacter Limnohabitans Lishizhenia Listeria Listonella Litoreibacter Litoribacillus Litoribacter Litoricola Litorilinea Litorimicrobium Litorimonas Litorisediminicola Loktanella Lonepinella Longilinea Longimycelium Longispora Lonsdalea Lucibacterium Vibrio Luedemannella Luminiphilus | Lutaonella Luteibacter Luteibaculum Luteimicrobium Luteimonas Luteipulveratus Luteivirga Luteococcus | 20 Luteolibacter Lutibacter Lutibaculum Lutimaribacter Lutimonas Lutispora Lysinibacillus Lysinimicrobium Lysinimonas Lysobacter Lyticum Macellibacteroides Macrococcus Macromonas Magnetococcus Magnetospira Magnetospirilum Magnetovibrio Mahella Malikia Malonomonas Mameliella Mangrovibacter Mangrovibacterium Mangroviflexus Mangrovimonas Mannheimia Maribacter Maribaculum Henriciella Maribius Maricaulis Marichromatium Maricurvus Marihabitans Marinactinospora Marinibacillus Jeotgalibacillus Marinicauda Marinicella Marinicola Roseivirga Marinifilum Mariniflexile Marinilabilia Marinilactibacillus Mariniluteicoccus Marinimicrobium Marininema Mariniradius Marinithermus ~~ Marinitoga Marinivirga Algibacter Marinobacter Marinobacterium Marinococcus Marinomonas Marinoscillum Marinospirillum Marinovum Mariprofundus Marisediminicola Marispirilum Maritalea Maritimibacter Maritimimonas Marivirga Marivita Marixanthomonas Marmoricola Martelella Marvinbryantia Massilia Mechercharimyces Megamonas Meganema Megasphaera Meiothermus Melaminivora Melghirimyces Melioribacter Melissococcus Melitea — synonym: Spongiibacter Melittangium Meniscus Meridianimaribacter | Mesoaciditoga Mesoflavibacter Mesonia Mesophilobacter Mesoplasma 78

ZELL

Mesorhizobium Mesotoga Metallibacterium Metallosphaera Metascardovia LU101096 Alloscardovia Methanimicrococcus Methanobacterium Methanobrevibacter Methanocalculus Methanocaldococcus Methanocella Methanococcoides Methanococcus Methanocorpusculum Methanoculleus Methanofollis Methanogenium Methanohalobium Methanohalophilus | Methanolacinia Methanolinea Methanolobus Methanomassilicoccus Methanomethylovorans Methanomicrobium Methanoplanus Methanopyrus Methanoregula Methanosaeta Methanosalsum Methanosarcina Methanosphaera Methanospirillum Methanothermobacter Methanothermococcus Methanothermus Methanothrix Methanosaeta Methanotorris Methermicoccus Methylarcula Methylibium Methylobacillus Methylobacter Methylobacterium Methylocaldum Methylocapsa Methyloceanibacter Methylocella Methylococcus Methylocystis Methyloferula Methylogaea Methylohalobius Methylohalomonas Methyloligella Methylomarinovum Methylomarinum Methylomicrobium | Methylomonas Methylonatrum Methyloparacoccus Methylophaga Methylophilus Methylopila Methylorhabdus Methylorosula Methylosarcina Methylosinus Methylosoma Methylosphaera Methylotenera Methylothermus Methyloversatilis Methylovirgula Methylovorus Methylovulum Micavibrio Microaerobacter Microbacterum Microbispora Microbulbifer Microcella Micrococcus Microcyclus Ancylobacter Microellobosporia Microlunatus Micromonas Micromonospora Micropolyspora Micropruina Microscilla Microsphaera Nakamurella Microterricola Microtetraspora Microvirga Microvirgula Millisia Miniimonas Mitsuaria Mitsuokella Mobilicoccus Mobiluncus Modestobacter Modicisalibacter Moellerella Mogibacterium Moheibacter Mongoliicoccus Mongoliitalea Mooreia Moorella Moraxella Morganella Moritella Morococcus Moryella Motilibacter Mucilaginibacter MucispirillumMumia Murdochiella Muricauda Muricoccus Roseomonas Muriicola Murinocardiopsis Myceligenerans Mycetocola Mycobacterium Mycoplana Mycoplasma Myroides Myxococcus Naasia Nafulsella Nakamurella Namhaeicola Nannocystis Natranaerobaculum Natranaerobius ~~ Natranaerovirga Natrialba Natribacillus ~~ Natrinema Natroniella Natronincola Natronoarchaeum Natronobacillus Natronobacterium Natronocella Natronococcus Natronoflexus Natronolimnobius Natronomonas Natronorubrum Natronovirga Naumannella Nautella Nautilia Naxibacter Massilia Necropsobacter Negativicoccus Neiella Neisseria Neoasaia | Neochlamydia Neokomagataea Neorickettsia Neptuniibacter Neptunomonas Nereida Nesiotobacter Nesterenkonia Nevskia Nguyenibacter Niabella 79

Niastella Nibrella Nibribacter Nicoletella Nisaea Nitratifractor Nitratireductor LU101096 Nitratiruptor Nitriliruptor Nitrincola Nitritalea Nitrobacter Nitrococcus Nitrolancea Nitrosococcus Nitrosolobus Nitrosospira Nitrosomonas Nitrosospira Nitrospina Nitrospira Nocardia Nocardioides Nocardiopsis Nonlabens Nonomuraea Noviherbaspirillum Novispirillum Novosphingobium Nubsella Obesumbacterium Oceanibacterium Oceanibaculum Oceanibulbus Oceanicaulis Oceanicella Oceanicola Oceanimonas Oceanirhabdus Oceaniserpentilla Oceanisphaera Oceanitalea Oceanithermus Oceanobacillus Oceanobacter Oceanococcus Oceanospirilum Oceanotoga Ochrobactrum Octadecabacter Odoribacter Oenococcus Oerskovia Ohtaekwangia Okibacterium Oleibacter Oleiphilus Oleispira Oligella Oligosphaera Oligotropha Olivibacter Olleya Olsenella Opitutus Orbus Orenia Oribacterium Oribaculum Porphyromonas Orientia Ornatilinea Ornithinibacillus Ornithinibacter ~~ Ornithinicoccus ~~ Ornithinimicrobium Ornithobacterium Oryzihumus Oscillibacter Oscillochloris Oscillospira Otariodibacter Ottowia Owenweeksia Oxalicibacterium Oxalobacter Oxalophagus Oxobacter Pacificibacter Paenalcaligenes Paenibacillus Paenirhodobacter Paenisporosarcina Paenochrobactrum Palaeococcus Palleronia Paludibacter Paludibacterium Panacagrimonas Pandoraea Pannonibacter Pantoea Papillibacter Parabacteroides Parachlamydia Paracoccus Paracraurococcus Paraeggerthella ~~ Paraferrimonas | Paraherbaspirilum Paralactobacillus Lactobacillus Paralcaligenes Paraliobacillus Paramoritella Paraoerskovia Parapedobacter Paraperlucidibaca Paraprevotella Parapusillimonas Pararhodobacter ~~ Pararhodospirilum Parascardovia Parasegetibacter Parasphingopyxis Parasporobacterium Parasutterella ParvibacterParvibaculum Parvimonas Parvularcula Pasteurella Pasteuria Patulibacter ~~ Paucibacter Paucimonas Paucisalibacillus ~~ Pectinatus Pectobacterium Pediococcus Pedobacter Pedomicrobium Pelagibaca Pelagibacillus Terribacillus Pelagibacterium Pelagibius Pelagicoccus Pelagicola Pelagimonas Pelczaria Pelistega Pelobacter ,Pelodictyon* Pelomonas Pelosinus Pelospora Pelotomaculum Peptococcus Peptoniphilus Peptostreptococcus Peredibacter Perexilibacter Perlucidibaca Persephonella Persicirhabdus Persicitalea Persicivirga Persicobacter Petrimonas Petrobacter Tepidiphilus Petrolinea Petrotoga Pfennigia Lamprocystis Phaeobacter Phaeochromatium Phaeocystidibacter Phaeospirillum Phaeovibrio Phascolarctobacterium Phaselicystis Phaseolibacter Phenylobacterium Phocaeicola Phocoenobacter Phorcysia Photobacterium Photorhabdus 80 Er

Phreatobacter Phycicoccus Phycicola Phycisphaera Phyllobacterium LU101096 Phytohabitans Phytomonospora Pibocella Picrophilus Pigmentiphaga Pilibacter Pilimelia Pillotina Pimelobacter Pirella Pirellula Piscibacillus Piscicoccus Pisciglobus Piscinibacter Piscirickettsia Planctomyces Planifilum Planktomarina Planktotalea Planobacterium Chryseobacterium Planobispora Planococcus Planomicrobium Planomonospora Planopolyspora Catenuloplanes Planosporangium Planotetraspora Plantactinospora Plantibacter Plasticicumulans Pleionea Pleomorphobacterium Pleomorphomonas Plesiocystis Plesiomonas Pluralibacter Polaribacter Polaromonas Polyangium Polycladomyces Polymorphobacter Polymorphospora Polynucleobacter Pontibaca Pontibacillus Pontibacter Ponticaulis Ponticoccus Pontimonas Pontirhabdus Algibacter Porphyrobacter Porphyromonas Porticoccus Poseidonocella Postechiella Pragia Prauserella Prevotella Pricia Primorskyibacter Prochloron Prochlorothrix Profundibacterium Prolinoborus Prolixibacter Promicromonospora Propionibacter Propionivibrio Propionibacterium Propionicicella Propioniciclava Propionicimonas Propioniferax Propionigenium Propionimicrobium Propionispira Propionispora Propionivibrio Prosthecobacter Prosthecochiloris Prosthecomicrobium Proteiniborus Proteiniclasticum Proteiniphilum Proteinivorax Proteocatella Proteus Protomonas Methylobacterium Providencia Pseudacidovorax Pseudahrensia Pseudaminobacter Pseudarcicella Pseudenhygromyxa Pseudidiomarina Idiomarina Pseudoalteromonas Pseudoamycolata Pseudonocardia Pseudobacteroides Pseudobutyrivibrio Pseudocaedibacter Pseudochrobactrum Pseudoclavibacter Pseudoduganella Pseudoflavonifractor Pseudofulvibacter Pseudofulvimonas Pseudogulbenkiania Pseudohaliea Pseudokineococcus Pseudolabrys Pseudomaricurvus Pseudomonas Pseudonocardia Pseudopedobacter Pseudoramibacter Pseudorhodobacter Pseudorhodoferax Pseudoruegeria Pseudosphingobacterium Pseudospirillum Pseudosporangium Pseudoteredinibacter Pseudothermotoga Pseudovibrio Pseudoxanthobacter Pseudoxanthomonas Pseudozobellia Psychrilyobacter _Psychrobacillus Psychrobacter Psychroflexus Psychroglaciecola Psychromonas Psychroserpens Psychrosphaera Pullulanibacillus Puniceibacterium Puniceicoccus Pusillimonas Pustulibacterium Pyramidobacter Pyrinomonas Pyrobaculum Pyrococcus Pyrodictium Pyrolobus Pyxidicoccus [Pyxicoccus] Quadricoccus Quatrionicoccus Quadrisphaera Quatrionicoccus Quinella 81

ZA

Rahnella Ralstonia Ramlibacter Raoultella Rapidithrix Rarobacter LU101096

Rathayibacter ~~ Rehaibacterium Reichenbachia Reichenbachiella Reichenbachiella Reinekea Renibacterium Reyranella Rhabdochromatium Rhabdothermus Rheinheimera Rhizobacter Rhizobium Rhizomicrobium Rhizomonas ~~ Sphingomonas Rhizorhabdus Rhizorhapis Rhodanobacter Rhodobaca Rhodobacter Rhodobium Rhodoblastus Rhodocista Rhodococcus

Rhodocyclus Rhodocytophaga Rhodoferax Rhodoglobus Rhodoligotrophos

Rhodomicrobium Rhodonellum Rhodopila Rhodopirellula Rhodoplanes

Rhodopseudomonas Rhodospira Rhodospirillum Rhodothalassium

Rhodothermus Rhodovarius Rhodovibrio Rhodovulum Rickettsia Rickettsiella Riemerella Rikenella Rivibacter Rivicola Robertkochia Robiginitalea

Robiginitomaculum Robinsoniella Rochalimaea Bartonella Romboutsia

Roseateles Roseburia Roseibaca Roseibacillus Roseibacterium Roseibium

Roseicitreum Roseicyclus Roseiflexus Roseimicrobium Roseinatronobacter

Roseisalinus Roseivirga Roseivivax | Rosenbergiella Roseobacter Roseococcus Roseomonas Roseospira Roseospirillum Roseovarius Rothia

Ruania Rubellimicrobium Rubribacterium Rubricoccus Rubrimonas

Rubritalea Rubritepida Rubrivirga Rubrivivax Rubrobacter Rudaea

Rudaeicoccus Rudaibacter Rudanella Ruegeria Rufibacter Rugamonas

Rugosimonospora Ruminobacter Ruminococcus Rummeliibacillus Runella Sabulilitoribacter Saccharibacillus Saccharibacter Saccharicrinis

Saccharobacter Saccharococcus Saccharofermentans Saccharomonospora

Saccharophagus Saccharopolyspora Saccharospirilum Saccharothrix

Sagittula Salana Salegentibacter Salibacillus Virgibacillus Salicola

Salimesophilobacter Salimicrobium Salinactinospora Salinarchaeum Salinarimonas Salinibacillus Salinibacter Salinibacterium Salinicoccus

Salinicola Salinigranum Salinihabitans Salinimicrobium Salinimonas

Salinirepens Salinirubrum Salinisphaera Salinispora Salinivibrio Salipiger

Salirhabdus Salisaeta Salisediminibacterium Saliterribacillus Salmonella15

Salsuginibacillus Samsonia Sandaracinobacter Sandaracinus Sandarakinorhabdus Sandarakinotalea Nonlabens Sanguibacter Saprospira

Sarcina Sarcobium Legionella Saxeibacter Nakamurella Scardovia Schineria

Ignatzschineria Schlegelella Schleiferia Schiesneria Schumannella

Schwartzia Sciscionella Sebaldella Sedimentibacter Sedimenticola

Sediminibacillus Sediminibacter Sediminibacterium Sediminicola Sediminihabitans Sediminimonas Sediminitomix Segetibacter Segniliparus

Seinonella Sejongia Chryseobacterium Selenihalanaerobacter

82 rs

Seleniivibrio Selenomonas Seliberia Senegalimassilia Seohaeicola LU101096 Seonamhaeicola Serinibacter Serinicoccus Serpens Serpula Brachyspira Serpulina Brachyspira Serratia Sharpea Shewanella Shigella Shimazuella Shimia Shimwellia Shinella Shivajiella Shuttleworthia Siansivirga Silanimonas Silicibacter Ruegeria Silvimonas Simiduia Simkania Simonsiella Simplicispira Singularimonas Solimonas Singulisphaera Sinibacillus Sinobaca Sinobacter Solimonas Sinobacterium Sinococcus ~~ Sinobaca Sinomicrobium Sinomonas Sinorhizobium Sinosporangium Siphonobacter Skermanella Skermania Slackia Smaragdicoccus Smithella Sneathia Sneathiella Snodgrassella Snuella Sodalis Soehngenia Solibacillus Solimonas Solirubrobacter Solitalea Solobacterium Soonwooa Sorangium Spelaeicoccus Sphaerisporangium Sphaerobacter Sphaerochaeta Sphaerosporangium Sphaerotilus Sphingobacterium Sphingobium Sphingomicrobium Sphingomonas Sphingopyxis Sphingorhabdus Sphingosinicella Spinactinospora Spirilliplanes = Spirillospora Spirillum Spirochaeta Spiroplasma Spirosoma Spongiibacter Spongiibacterium Spongiimonas Spongiispira Sporacetigenium Sporanaerobacter Sporichthya Sporobacter Sporobacterium | Sporocytophaga Sporohalobacter Sporolactobacillus Sporolituus Sporomusa Sporosalibacterium Sporosarcina Sporotalea Pelosinus Sporotomaculum Stackebrandtia Stakelama Staleya Sulfitobacter Stanierella Aquimarina Staphylococcus Staphylothermus Stappia Starkeya Stella Stenothermobacter Nonlabens Stenotrophomonas Stenoxybacter Steroidobacter Sterolibacterum Stetteria Stibiobacter Stigmatella Stomatobaculum Stomatococcus Rothia Streptacidiphilus Streptoalloteichus Streptobacillus Streptococcus Streptohalobacillus Streptomonospora Streptomyces Streptosporangium Streptoverticillium Streptomyces Stygiolobus Subdoligranulum Subsaxibacter Subsaximicrobium Subtercola Succinatimonas Succiniclasticum Succinimonas Succinispira Succinivibrio Sulfitobacter Sulfobacillus Sulfolobus Sulfophobococcus Sulfuricella Sulfuricurvum Sulfurihydrogenibium Sulfurimonas Sulfurisoma Sulfurisphaera Sulfuritalea Sulfurivirga Sulfurococcus Sulfurospirillum Sulfurovum Sungkyunkwania Sunxiuginia Sutterella Suttonella Swaminathania Swingsia Symbiobacterium Symbiotes Synergistes Syntrophaceticus | Syntrophobacter ~ Syntrophobotulus ~~ Syntrophococcus Syntrophomonas Syntrophorhabdus Syntrophospora Syntrophomonas Syntrophothermus Syntrophus Tabrizicola Tahibacter Taibaiella Tamlana Tamlicoccus Tannerella Tanticharoenia Taonella Tardiphaga Tateyamaria 83

Tatlockia Tatumella Taylorella Tectibacter Teichococcus Roseomonas Telluria 1101096 Telmatobacter Telmatocola Telmatospirillum Tenacibaculum Tenuibacillus Tepidamorphus Tepidanaerobacter Tepidibacillus Tepidibacter Tepidicella Tepidimicrobium Tepidimonas Tepidiphilus Terasakiella Teredinibacter Terrabacter Terracoccus Terribacillus Terriglobus Terrimicrobium Terrimonas Terrisporobacter Tersicoccus Tessaracoccus Tetragenococcus Tetrasphaera Tetrathiobacter Advenella Texcoconibacillus Thalassobacillus Thalassobacter Thalassobaculum Thalassobius Thalassococcus Thalassolituus Thalassomonas Thalassospira Thalassotalea Thauera Thermacetogenium Thermaerobacter Thermanaeromonas Thermanaerovibrio Thermasporomyces Thermicanus Thermincola Thermithiobacillus Thermoactinomyces Thermoactinospora Thermoanaerobacter Thermoanaerobacterium Thermoanaerobaculum Thermoanaerobium Thermobacillus Thermobacteroides Thermobifida Thermobispora Thermobrachium Thermocatellispora Thermochromatium Thermocladium Thermococcoides Kosmotoga Thermococcus Thermocrinis Thermocrispum Thermodesulfatator Thermodesulfobacterium Thermodesulfobium Thermodesulforhabdus Thermodesulfovibrio Thermodiscus Thermofilum Thermoflavifilum Thermoflavimicrobium Thermoflexus Thermogemmatispora Thermogymnomonas Thermohalobacter Thermohydrogenium Thermoleophilum Thermolithobacter Thermolongibacillus Thermomarinilinea Thermomicrobium Thermomonas Thermomonospora Thermonema Thermophagus Thermoplasma Thermopolyspora Thermoproteus Thermosediminibacter Thermosinus Thermosipho Thermosphaera Thermosporothrix Thermosulfidibacter Thermosulfurimonas Thermosyntropha Thermotalea Thermoterrabacterium Carboxydothermus Thermothrix Thermotoga Thermotunica Thermovenabulum Thermovibrio Thermovirga Thermovorax Thermovum Thermus Thioalbus Thioalkalibacter ~~ Thioalkalicoccus Thioalkalimicrobium Thioalkalispira Thioalkalivibrio Thiobaca Thiobacillus Thiobacter Thiobacterium Thiocapsa Thioclava Thiococcus Thiocystis Thiodictyon Thiofaba Thioflavicoccus Thiohalobacter Thiohalocapsa Thiohalomonas Thiohalophilus Thiohalorhabdus Thiohalospira Thiolamprovum Thiomargarita Thiomicrospira Thiomonas Thiopedia Thiophaeococcus Thioploca Thioprofundum Thioreductor Thiorhodococcus Thiorhodospira Thiorhodovibrio Thiosphaera Paracoccus Thiospira Thiospirillum Thiothrix Thiovirga Thiovulum Thorsellia Tindallia Tissierella Tistlia Tistrella 84

Tolumonas Tomitella Tonsilliphilus Toxothrix Trabulsiella Tranquillimonas LU101096 Treponema Trichlorobacter Geobacter Trichococcus Tropheryma Tropicibacter Tropicimonas Truepera Trueperella Tsukamurella Tuberibacillus Tumebacillus Turicella Turicibacter Turneriella Uliginosibacterium Ulvibacter Umboniibacter Umezawaea Undibacterium Ureaplasma Ureibacillus Uruburuella Vadicella Vagococcus Vallitalea Vampirovibrio Varibaculum Variovorax Vasilyevaea Veillonella Venenivibrio Verminephrobacter Verrucomicrobium Verrucosispora Vibrio Vibrionimonas Victivallis Virgibacillus Virgisporangium Viridibacillus Vitellibacter Vitreoscilla Vogesella Volcaniella Halomonas Volucribacter Vulcanibacillus Vulcaniibacterum Vulcanisaeta Vulcanithermus Waddlia Wandonia Wangia Zunongwangia Wautersia Cupriavidus Wautersiella Weeksella Weissella Wenxinia Wenyingzhuangia Wigglesworthia Williamsia Winogradskyella Wohlfahrtimonas Wolbachia Wolinella Woodsholea Xanthobacter Xanthomonas Xenophilus Xenorhabdus Xiangella Xylanibacter Prevotella Xylanibacterium Xylanimicrobium Xylanimonas Xylella Xylophilus Yangia Yania Yaniella Yaniella Yeosuana Yersinia Yimella Yokenella — synonym: Koserella Yonghaparkia Youngiibacter Yuhushiella Zavarzinella Zavarzinia Zeaxanthinibacter Zhangella Maritalea Zhihengliuella Zhongshania Zhouia Zimmermannella = Pseudoclavibacter Zobellella Zobellia Zoogloea Zooshikella Zunongwangia Zymobacter Zymomonas Zymophilus: iii) said microorganism is non-pathogenic and/or opportunistic pathogen and is selected from the group consisting of genera as defined in item (ii) herein; iv) said microorganism is Gram-positive, non-pathogenic and/or opportunistic pathogen and is selected from the group consisting of genera as defined in item (ii) herein; v) a fungus; preferably said fungus is selected from the group consisting of: Candida yeasts, Saccharomyces yeasts and yeasts in the family Dipodascaceae; further preferably said Dipodascaceae yeasts are Galactomyces, Geotrichum or Saprochaete yeasts, most preferably said Saccharomyces yeast is Saccharomyces boulardii, S. cerevisiae, S. pastorianus, or Schizosaccharomyces pombe; vi) a protozoa, said protozoa being non-pathogenic to human, preferably said protozoa is Chilomastix mesnili, Endolimax nana, Entamoeba coli, Entamoeba dispar, Entamoeba hartmanni, Entamoeba polecki or lodamoeba buetschiii. 85

32. The method for producing or isolating a modified microorganism according to any one LU101096 of preceding items, wherein said microorganism is selected from genera consisting of: Lactobacillus, Bifidobacterium, Bacteroides and Enterococcus, wherein said culture medium is BSM or SBSM with or without propionic acid, preferably said microorganism is Lactobacillus paracasei or Lactobacillus reuteri.

33. The method for producing or isolating a modified microorganism according to any one of preceding items, wherein the specific combination of said microorganism, culture medium (i.e., growth medium), growth temperature, temperature of adding of said sol- ubilized heterologous lipid carrier (i.e., loading temperature), pH and loading medium are selected from the group consisting of the following combinations: Microorganism Growth Approx. Approx. Approx. | Loading Medium growth T | loading T | loading | Medium (e.g. (e.g. pH “about” “about” (e.g., value °C) | value °C) | “about” pH value) Lactobacillus MRS 30°C or | 30°C, 6or7 PBS or BSM paracasei or 37°C 37°c or with PBS 46°C eg. 1to 1 Lactobacillus (eg. 1 to ; ratio) or reuteri BSM or

SBSM Lactobacillus MRS with | 30°C or | 30°C, 6or7 PBS or BSM paracasei or propionic 37°C 37°c or with PBS cid e.g. 46°C .g., 110 1 Lactobacillus ac (eg (e.g © ; 5ml/L) ratio) or reuteri BSM or

SBSM Lactobacillus BSM OR | 30°C, 30°C, 4, 5, 6 | BSM with paracasei or SBSM 37°C, 37°C, or 7 PBS (e.g., 1 | 46°C or|46°C or to 1 ratio) or Lactobacillus ; 55°C 55°C BSM or reuteri

SBSM 86

Lactobacillus BSM OR | 30°C, 30°C, 4, 5, 6 | BSM with LU101096 paracasei or SBSM with | 37°C, 37°C, or 7 PBS (e.g., 1 Lactobacillus propionic 46°C or|46°C or to 1 ratio) or | acid (e.g. | 55°C 55°C BSM or reuteri 5m//L) SBSM Bifidobacterium BSM OR | 30°C, 30°C, 6or7 | BSM with breves SBSM 37°C, 37°C, PBS (e.g., 1 46°C to 1 ratio) or BSM or

SBSM Bifidobacterium BSM OR | 30°C, 30°C, 6or7 | BSM with breves SBSM with | 37°C, 37°C, PBS (e.g., 1 propionic 46°C to 1 ratio) or acid (e.g. BSM or 5ml/L) SBSM Bifidobacterium BSM OR | 30°C, 30°C, 6or7 | BSM with longum SBSM 37°C, 37°C, PBS (e.g., 1 46°C to 1 rati Or Bifidobacterium 0 1 ratio) or ; ; BSM or infantis

SBSM Bifidobacterium BSM OR | 30°C, 30°C, 6or7 | BSM with longum SBSM with | 37°C, 37°C, PBS (e.g., 1 ioni 46°C to 1 ratio) o Or Bifidobacterium propionic lo) or i , acid (e.g. BSM or infantis 5ml/L) SBSM Bacteroides WC, BSM | 30°C, 30°C, 6or7 | BSM with vulgatus ORSBSM | 37°C, 37°C, PBS (e.g., 1 | 46°C to 1 ratio) or BSM or

SBSM Enterococcus MRS 30°C or | 30°C, 6or7 PBS or BSM faecalis 37°C 37°c or with PBS 46°C (e.g., 1 to 1 ratio) or 87

BSM or LU101096

SBSM Enterococcus MRS with | 30°C or! 30°C, 6or7 PBS or BSM faecalis propionic 37°C 37°c or with PBS acid (e.g. 46°C (e.g., 1 to 1 Smi/L) ratio) or BSM or

SBSM Enterococcus BSM OR | 30°C, 30°C, 4, 5, 6 | BSM with faecalis SBSM 37°C, 37°C, or 7 PBS (e.g., 1 46°C or to 1 ratio) or 55°C BSM or

SBSM Enterococcus BSM OR | 30°C, 30°C, 4, 5, 6 | BSM with faecalis SBSM with | 37°C, 37°C, or 7 PBS (e.g., 1 propionic 46°C or to 1 ratio) or acid (e.g, 55°C BSM or 5ml/L) SBSM

34. The method for producing or isolating according to any one of preceding items, wherein said microorganism is the microorganism according to any one of preceding items.

35. A modified microorganism comprising: i) a cell and ii) a heterologous lipid carrier, said lipid carrier comprising: a) a lipid portion, wherein said lipid portion is at least partially associated with an exterior surface of said cell of said modified microorganism, wherein said lipid portion comprising: a ceramide-like glycolipid moiety (and/or a ceramide moiety and/or a sphingolipid moiety and/or sphingosine moiety) and/or a fatty acid moiety; preferably said exterior surface of said cell comprising: a cell wall and/or a cell membrane and/or an outer cell membrane and/or a polysaccharide (e.g., capsule polysaccharide); further preferably said lipid portion is at least partially 88incorporated and/or adhered and/or bound to said exterior surface of LU101096 said cell; and b) a non-lipid portion, wherein said microorganism is capable of locating and/or displaying said non-lipid portion or fragment thereof onto the exterior surface of said cell, preferably said non-lipid portion comprising a carbohydrate moiety and/or lipopeptide moiety (e.g., a glycosylated lipopeptide moiety, e.g., a carbohydrate antigen); further preferably said non-lipid portion is capable of binding and/or reducing toxicity and/or neutralizing a toxin; iii) optionally, said modified microorganism (e.g., said lipid carrier) further comprising a heterologous steroid moiety; preferably said steroid moiety is cholesterol, a derivative or analogue thereof; produced or isolated by the method for producing or isolating according to any one of preceding items.

36. A method for treatment, amelioration, prophylaxis or diagnostics of a disease, said method comprising: i) providing one or more (e.g., mixtures) microorganism, composition, vaccine or adjuvant according to any one of preceding items to a subject (e.g., patient) in need thereof (e.g., human or animal); ii) administering a therapeutically effective amount of said microorganism, composition, vaccine or adjuvant to said subject (e.g., patient).

37. A method for eliciting or modulating an immune response, said method comprising: i) providing the microorganism, composition, vaccine or adjuvant according to any one of preceding items to a subject (e.g., patient) (e.g., human or animal); ii) administering a therapeutically effective amount of said microorganism, composition, vaccine or adjuvant to said subject (e.g., patient).

38. A method for screening for a lipid carrier, growth medium, loading medium, loading conditions (e.g., of the lipid carrier to the microorganism) or growth conditings, said method comprising: i) providing the microorganism, composition, vaccine or adjuvant according to any one of preceding items; 89ii) providing a lipid carrier; LU101096 iii) exposing (e.g., adding or loading) said lipid carrier to said microorganism, composition, vaccine or adjuvant according to any one of preceding items under suitable conditions; iv) optionally, evaluating the effect of the lipid portion (e.g., a fatty acid or ceramide moiety) and/or the non-lipid portion (e.g., polypeptide) of said lipid carrier on association (e.g., stability, strength, amount and/or concentration) between said lipid carrier with said microorganism, composition, vaccine or adjuvant according to any one of preceding items; Vv) optionally, carring out said method under different condition, preferably with another growth medium and/or loading medium and/or under different loading conditions (e.g., of the lipid carrier to the microorganism) and/or growing conditings (e.g., temperature and/or pH) (e.g., different from those according to any one of preceding items) and evaluating the effect of said condition (preferably another growth medium and/or loading medium and/or under different loading conditings (e.g., temperature and/or pH)) on the association (e.g., strength, stability, concentration and/or amount) of said lipid carrier with said microorganism.

39. The microorganism, composition, vaccine or adjuvant according to any one of preceding items, for use as a medicament.

40. The microorganism, composition, vaccine or adjuvant according to any one of preceding items, for use in one or more of the following methods: i) in a method for treatment, amelioration, prophylaxis or diagnostics of a disease (e.g., a bacterial, viral, or fungal infection or autoimmune diseases, e.g., suitable for topical applications to skin and/or other sites such urogenital tract, respiratory system or oral cavity); ii) in a method for modulating the mammalian immune response; iii) in a method for eliciting or modulating an immune response; iv) in a method for monitoring development of a disease and/or assessing the efficacy of a therapy of a disease; v) in a method for screening a candidate compound for activity against a disease; vi) in a method for delivering a pharmaceutically active compound, preferably said delivering is to a mucosal tissue of a subject (e.g., patient); 90vii) in a method for vaccine delivery; LU101096 viii)in a method for screening of microorganisms capable of binding the lipid carrier according to any one of preceding items; ix) in a method for binding and/or reducing toxicity and/or neutralizing a toxin; x) in a method for binding a receptor of a toxin; xi) in a method for binding and/or reducing the pathogenicity and/or neutralizing a pathogenic microorganism; xii) in a method for binding a receptor of a pathogenic microorganism; xiii)in a method for producing or isolating the microorganism according to any one of preceding items; xiv)in a method according to any one of preceding items.

41. A use of the microorganism, composition, vaccine or adjuvant according to any one of preceding items for one or more of the following: i) for treatment, amelioration, prophylaxis or diagnostics of a disease (e.g., a bacterial, viral, or fungal infection or autoimmune diseases, e.g., suitable for topical applications to skin and/or other sites such urogenital tract, respiratory system or oral cavity); ii) for modulating the mammalian immune response; iii) for eliciting or modulating an immune response; iv) for monitoring development of a disease and/or assessing the efficacy of a therapy of a disease; v) for screening a candidate compound for activity against a disease; vi) for delivering a pharmaceutically active compound, preferably said delivering is to a mucosal tissue of a subject (e.g., patient); vii) for vaccine delivery; viii) for screening of microorganisms capable of binding the lipid carrier according to any one of preceding items; ix) for binding and/or reducing toxicity of and/or neutralizing a toxin; x) for binding a receptor of a toxin; 91 |xi) for binding and/or reducing the pathogenicity and/or neutralizing a pathogenic LU101096 microorganism; xii) for binding a receptor of a pathogenic microorganism; xiii) for producing or isolating the microorganism according to any one of preceding items; xiv)use in a method according to any one of preceding items.

42. The microorganism, composition, vaccine, adjuvant, method or use according to any one of preceding items, wherein said toxin is selected from the group consisting of: heat labile toxin (LT), heat stabile toxin (ST), Vero-toxins or Shiga-like toxins (Stxs), cytotoxins, endotoxins (LPS), Enteroaggregative ST toxin (EAST), Shiga toxin (STxs), Shigella enterotoxins 1 (ShET1), Shigella enterotoxins 2 (ShET2), neurotoxin, cytolethal distending toxins (Cdt), Salmonella typhimurium AvrA toxin, cytotoxic necrotizing factor (CNFy), Yersinia murine toxin (Ymt), Yersinia enterocolitica Yst toxin, toxin complex (TCa), heat stabile toxin, Enterobacter cloacae leukotoxin, Shiga-like toxin Il, heat-stable like enterotoxins, extracellular toxic complex (ETC), hemolysins (Shi), pore-forming Toxin (PFT), a-hemolysin (HIyA), heat-stable like toxin, cytotoxins, Clostridium perfringens alpha-toxin (CpPLC), Clostridium perfringens beta toxin, Clostridium perfringens enterotoxin (CPE), Clostridium difficile enterotoxins (Tcd), Clostridium botulinum neurotoxins, Clostridium tetani tetanospasmin, Clostridium botulinum C2 toxin, Clostridium botulinum C3 toxin, Clostridium perfringens epsilon- toxin (e-toxin), Clostridium perfringens iota-toxin (i-toxin), tetanus neurotoxin (TeNT), theta-toxin (Perfringolysin O), Clostridium spiroforme (spiroforme toxin), Clostridium septicum (a-toxin), Lecithinase, Cholera toxins (CTx), accessory cholera enterotoxin (Ace), RTX toxin, Vibrio cholerae Zonula occludens toxin (Zot), Vibrio cholerae Cholix toxin, a-hemolysin, B-hemolysin, 5-hemolysin, A-hemolysin, Staphylococcus aureus exfoliative toxins (Exofoliatins), Panton-Valentine leukocidin (PVL), staphylococcal enterotoxins (SE), Toxic shock syndrome toxin-1 (TSST-1), B-hemolysin/cytolysin, CAMP factor, streptolysin O, streptolysin S, pneumolysin (PLY), Streptococcus pyogenes Exotoxins (PSE), Helicobacter pylori vacuolating cytotoxin A (VacA), cytolytic toxins, Pseudomonas aeruginosa Exotoxins (ExoA, ExoS, ExoT, ExoU, ExoY), Phospholipase C (PLC), Pasteurella multocida Toxin (PMT), RTX toxins, Bacillus weihenstephanensis endotoxins, Bacillus cereus hemolysin BL (HBL), Bacillus cereus non-hemolytic enterotoxin (Nhe), Bacillus cereus Cytotoxin K (CytK), Bacillus cereus | emetic toxin, Bacillus cereus toxin (cereolysin), Bacillus anthracis (Anthrax toxin), Bacillus thuringiensis 5-endotoxins (Cry toxins), Cytolethal distending toxin (cdtA, cdtB, 92cdtC), cholera-like enterotoxin, Aeromonas hydrophila aerolysin-related cytotoxic LU101096 enterotoxin (Act), ADP-ribosylation toxin, a-hemolysins, b-hemolysins, heat labile toxin (LT+), heat stabile toxin (ST+), endotoxins (LPS), Bordetella pertussis pertussis toxin, Adenylate cyclase toxin, Tracheal cytotoxin, dermonecrotic toxin (heat-labile), endotoxins (LPS), Endotoxin (LOS), Haemophilus ducreyi cytolethal distending toxins (HdCDT), Hemolysins, Endotoxins, Cytotoxins, Diphtheria Toxin, Exotoxins, Bacteroides fragilis toxin (bft), Listeria monocytogenes Listeriolysin O, or Rotavirus toxin NSP4.

43. The microorganism, composition, vaccine, adjuvant, method or use according to any one of preceding items, wherein said disease is selected from the group consisting of: infectious disease, gastrointestinal disorder, inflammatory bowel disease, Celiac Disease, cancer, gastrointestinal cancer, Gastrointestinal adenocarcinoma, inflammatory disease, auto-immune disease, poisoning, foodborne poisoning, allergy, parasitic disease, intestinal infectious diseases (e.g., Cholera due to Vibrio cholerae 01, biovar cholerae; Cholera due to Vibrio cholerae 01, biovar eltor; Cholera); Typhoid and paratyphoid fevers; Infection due to Salmonella typhi; Paratyphoid fever A; Paratyphoid fever B; Paratyphoid fever C; Paratyphoid fever; Infection due to Salmonella paratyphi; Salmonella infections; infection or foodborne intoxication due to any Salmonella species other than S. typhi and S. paratyphi; Salmonella enteritis; Salmonellosis; Salmonella sepsis; Localized salmonella infections; Salmonella arthritis; Salmonella meningitis; Salmonella osteomyelitis; Salmonella pneumonia; Salmonella renal tubulo-interstitial disease; Salmonella infection; Shigellosis; Shigellosis due to Shigella dysenteriae; Group A shigellosis; Shiga-Kruse dysentery; Shigellosis due to Shigella flexneri; Group B shigellosis; Shigellosis due to Shigella boydii; Group C shigellosis; Shigellosis due to Shigella sonnei; Group D shigellosis; Bacillary dysentery; bacterial intestinal infections; Enteropathogenic Escherichia coli infection; Enterotoxigenic Escherichia coli infection; Enteroinvasive Escherichia coli infection; Enterohaemorrhagic Escherichia coli infection; intestinal Escherichia coli infections; Escherichia coli enteritis; Campylobacter enteritis; Enteritis due to Yersinia enterocolitica; Enterocolitis due to Clostridium difficile; Foodborne intoxication by Clostridium difficile; Pseudomembranous colitis; Bacterial enteritis; Foodborne staphylococcal intoxication; Botulism; foodborne intoxication due to Clostridium botulinum: Foodborne Clostridium perfringens intoxication; Foodborne Clostridium welchii intoxication; Enteritis necroticans; Pig-bel; Foodborne Vibrio parahaemolyticus intoxication; Foodborne Bacillus cereus intoxication; Bacterial foodborne intoxication; Amoebiasis; infection due to Entamoeba histolytica; Acute amoebic dysentery; Acute amoebiasis; Intestinal amoebiasis; Chronic intestinal amoebiasis; Amoebic 93nondysenteric colitis; Amoeboma of intestine; Amoeboma; Amoebic liver abscess; LU101096 Hepatic amoebiasis; Amoebic lung abscess; Amoebic abscess of lung (and liver); Amoebic brain abscess; Amoebic abscess of brain (and liver)(and lung); Cutaneous amoebiasis; Amoebic appendicitis; Amoebic balanitis; Amoebiasis; protozoal intestinal diseases; Balantidiasis; Balantidial dysentery; Giardiasis; lambliasis; Cryptosporidiosis; Isosporiasis; Infection due to Isospora belli and Isospora hominis; Intestinal coccidiosis: Isosporosis; Intestinal trichomoniasis; Sarcocystosis; Sarcosporidiosis; Flagellate diarrhoea; Protozoal colitis, Protozoal diarrhea; Protozoal dysentery; Viral intestinal infections; Rotaviral enteritis; Acute gastroenteropathy due to Norwalk agent; Small round structured virus enteritis; Adenoviral enteritis; viral enteritis; Viral intestinal infection; Viral enteritis; Viral gastroenteritis; Viral gastroenteropathy; gastroenteritis and colitis of infectious origin; Catarrh, enteric or intestinal; Diarrhoea acute bloody; Diarrhoea acute haemorrhagic; Diarrhoea acute watery; Diarrhoea dysenteric; Diarrhoea epidemic; Infectious or septic colitis, enteritis or gastroenteritis; Neonatal diarrhoea.

44. The microorganism, composition, vaccine, adjuvant, method or use according to any one of preceding items, wherein said pathogenic microorganism is any pathogenic microorganism according to any preceding items.

45. The microorganism, composition, vaccine, adjuvant, method or use according to any one of preceding items, wherein said microorganism is isolated and/or live and/or killed and/or attenuated and/or pasteurized and/or lyophilized and/or freeze dried.

46. The microorganism, composition, vaccine, adjuvant, method or use according to any one of preceding items, wherein said microorganism is capable to stably comprising said lipid carrier under one or more of the following conditions: i) pasteurization conditions; ii) freeze and thaw conditions; ii) lyophilization conditions; iv) incubation at pH of about 1 for about 1 hour at about 37°C; V) incubation for up to about 4 hours at about 37°C; vi) incubation in the presence of gastric juice for about 30 min at about 37°C; vii) storage at about 4°C for up to about 4 weeks. 94

47. The microorganism, composition, vaccine, adjuvant, method or use according to any LU101096 | one of preceding items, wherein said method or use is an in vitro, ex vivo or in vivo method or use.

48. A kit comprising the microorganism, composition, vaccine or adjuvant according to any one of preceding items.

49. A kit for performing a method according to any one of preceding items. Examples of the invention

[00116] In order that the invention may be readily understood and put into practical effect, some aspects of the invention are described by way of the following non-limiting examples.

[00117] Example 1: Use of GM1 molecule as a receptor for the Cholera Toxin (CT)

[00118] Monosialoganglioside (GM1) is a glycosphingolipid composed of a ceramide (Sphingosine and Fatty Acid) and an oligosaccharide. The oligosaccharide part of GM1 is known to be a natural receptor for the cholera toxin (CT). Cholera is an acute, diarrheal illness caused by infection of the intestine with the bacterium Vibrio cholerae. An estimated 3 to 5 million cases and over 100,000 deaths occur each year around the world. Heat labile toxin (LT) from ETEC and Cholera toxin are highly identical, and both were reported to bind the same carbohydrate receptor GM1 described above. However, slightly different specificities were reported. A non-pathogenic microorganism loaded with the GM1 and presenting the oligosaccharide portion of its structure to the environment (i.e., an exterior surface of the cell) may be delivered directly to the gastrointestinal tract where it may bind CT, thereby abating, curing, treating or preventing the development of the Vibrio cholerae- associated diseases. Such GM1-loaded microorganisms would overcome most of the drawbacks presented by other available therapeutics.

[00119] Material and Methods

[00120] L. Coupling the Cholera Toxin to magnet beads:

[00121] 1.1. Preparation of beads

[00122] Dynabeads® M-270 Amine (Life technologies, UK) were used to directly link the Cholera toxin (List Biological Laboratories, Inc). The Surface-reactive primary amino- groups allow immobilization of ligands such as carbohydrates, glycoproteins and glycolipids through reductive amination of aldehyde or ketone groups. Alternatively, ligands can be immobilized through amide-bond formation with carbodiimide-activated carboxylic acid groups. Bi-functional cross-linkers may be used to introduce other functional groups. 95

[00123] 25 yl of Dynabeads® M-270 Amine were placed in 1.5 ml Safe-Lock | U101096 Eppendorf tubes (Eppendorf, Hamburg, Germany), washed twice with 100 pl buffer F (0.26 g NaH2PO4+H20, 1.44 g Na2HPO4+2H20, 8.78 g NaCl, pH 7.4) using the Dynal Magnetic Particle Concentrator®-S (MPC®-S, Dynal Biotech, Oslo, Norway) and suspended in 25 pl buffer F. Before coupling of ligands to Beads the washed Dynabeads were activated with NHS (N-hydroxy-succinimidyl)-ester cross-Linker. Hereby the beads were re-suspended in

0.1 M sodium phosphate buffer with 0.15 M NaCl, pH 7.4. Dissolved NHS-ester (15 mg/ml) were added to the bead-solution. Beads were then incubated for 30 min at room temperature with slow tilt and rotation, and finally washed twice with buffer F and re-suspended in 25 pi buffer F. Cholera toxin were then dissolved in water according to the manufacturer's recommendations (List Biological Laboratories, California, USA). To eliminate amino groups (Tris-) present in the initial buffer, Cholera toxin solution were dialyzed against PBS.

[00124] 1.2. Preparation and GM1-loading of samples

[00125] 1g of a mixture of microorganisms (e.g., fresh feces sample from healthy adults) were suspended in a tube containing 9 ml of anaerobic PBSred (8.5 g/l NaCl, 0.6 g/l Na2HPO4, 0.3 g/l KH2PO4, 0.25 g/l Cystein.HCI, 0.1 g/l Peptone, pH 7,0) and stored at 4°C in an anaerobic box (Anaerogen, Oxoid, Wesel, Germany). The samples were then processed as follows: sterile 3 mm diameter glass beads were added, and the samples homogenized by vortexing. The homogenized fecal suspensions were centrifuged (300 x g for 1 min) to sediment debris. The resulting supernatants were transferred in new tubes and centrifuged again. The supernatants were diluted 1:100 (v/v) in different growth media (i.e., LB broth, MRS broth and WC broth) and incubated overnight at 37°C under anaerobic conditions. The cultures were washed with PBS, resuspended in anaerobic PBS and used directly or frozen at -20°C for later use. The cultures in PBS were set to an OD1 to 10 added with 1 to 10ug/ml of GM1 (carbosynth) and incubated at 30°C or 37°C overnight. The cells were than extensively washed with PBS before being resuspended in PBSred 0.1% HSA.

[00126] Optionally, the fecal sample may be depleted or enriched for any specific genus or species of microorganisms, e.g., by mean of affinity depletion/enrichment, antibiotic treatment or any suitable alternative method known in the art. For example, in order to increase the proportion of Bifidobacterium and Lactobacillus microorganisms (bacteria) in fecal samples, the samples were further depleted for Bacteroides (bacteria) by the means of centrifugations. Bacteroides are the most abundant genus of the human colonic microbiota (i.e., microflora), surpassing Lactobacillus and Bifidobacterium by a factor of 10.000. In order to increase the proportion of Bifidobacterium and Lactobacillus bacteria in fecal samples, the feces suspensions were centrifuged for 3 min at 2500 rpm. Due to their small size, Bacteroides were mostly restricted to the supernatant that could be discarded or used for 96further experiments. The pellets were resuspended in PBSred and the procedure was LU101096 repeated three times. The final bacteria pellets were loaded with GM1 as described above.

[00127] Il. Isolation of microorganisms naturally binding GM1 and presenting the glycosylic moiety to the environment (i.e., on their exterior surface)

[00128] 1.1. Isolation of bacteria using toxin-coated beads

[00129] 20 ul of GM1-loaded samples were added to 175ul PBSred + 0.1% HSA and 5 ul “Cholera toxin”-coated beads. The mixes were incubated at a suitable temperature (e.g., room temperature (RT) or 37°C) under appropriate atmosphere and gentle agitation for 1 hour. Subsequently, the beads were washed twice with 200pl PBSred + 0.01% HSA. To reduce non-specific binding of viable bacteria, the beads were pre-incubated for 1 hour with pasteurized bacteria (e.g., at RT or 37°C). Pasteurized bacteria may also be added during the incubation of viable bacteria with the heat labile toxin-coated beads.

[00130] After incubation the beads were washed and resuspended in 1ml PBSred. 100 ul aliquots were spread-plated on unspecific and specific agar plates like, e.g., WC agar (Oxoid, Wesel, Germany), MRS agar (Roth, Karlsruhe, Germany), Bifidus Selective agar (BSM, Sigma Aldrich, Taufkirchen, Germany) and Self made Bifidobacterium Selective agar.

[00131] WC Medium: Tryptone 10g/L; Gelatin peptone 10g/L; Yeast extract 5g/L; Glucose 1g/L; Sodium chloride 5g/L; L-Arginine 1g/L; Sodium pyruvate 1g/L; Menadione

0.0005g/L; Haemin 0.005 g/L (pH=7+0.2).

[00132] WC Agar: Tryptone 10g/L; Gelatin peptone 10g/L; Yeast extract 5g/L; Glucose 1g/L; Sodium chloride 5g/L; L-Arginine 1g/L; Sodium pyruvate 1g/L; Menadione 0.0005g/L; Haemin 0.005 g/L; Agr 10g/L (pH=7+0.2).

[00133] MRS Agar: Peptone 10 g/l; Yeast extract 4 g/l; Beef extract 8 g/l; Glucose 20 g/l; Dipotassium phosphate 2 g/l; Sodium acetate 5 g/l, Ammonium citrate 2 g/l, Magnesium sulphate (MgSO,) 0.2 g/l; Manganese sulphate 0.05 g/l; Polysorbate 80 1 g/l; Agar 10 g/l; pH= 6.2 +0.2.

[00134] BSM Agar: (Sigma Aldrich Catalog Number in May 2018: 88517). Principle and | Interpretation. BSM contains Peptone and Meat extract as sources of carbon, nitrogen, | vitamins and minerals. Yeast extract supplies B-complex vitamins which stimulate bacterial growth. Dextrose is the carbohydrate source. Sodium chloride maintains the osmotic balance. There is a compound in low concentration for detoxify metabolic by-products. The medium contains reducing and buffering agents. Selective salts inhibit the growth of molds, Enterococci and other Gram-negative bacteria. Another compound inhibits glycolysis by inactivating glyceraldehyde-3-phosphate dehydrogenase present and important in different bacteria and fungi (also Streptococci sp.). Three antibiotics are the selective agents and 97inhibit the accompanying bacterial flora like Bacilli, Enterobacteriaceae and Pseudomonas. LU101096 Bifidobacteria that can reduce an azo compound present in the medium, which gives the colonies a pink-purple coloration.

[00135] SBSM agar contains casein peptone (12 g/l), meat peptone (5 g/l), sodium chloride (5 g/l), beef extract (3 g/l), yeast extract (3 g/l), cornstarch (19/1), glucose (2.5 g/l), lactulose (2.5 g/l), cysteine-hydrochloride (0.5 g/l), riboflavin (0.01 g/l), propionic acid (99%; Sm//l) and Agar 10g/l. All components were mixed and suspended in distilled water. The pH was adjusted to 5.5 +/- 0.2 with 5N NaOH.

[00136] BSM Broth: (Sigma Aldrich Catalog Number in May 2018: 90273). Principle and Interpretation: BSM contains Peptone and Meat extract as sources of carbon, nitrogen, vitamins and minerals. Yeast extract supplies B-complex vitamins which stimulate bacterial growth. Dextrose is the carbohydrate source. Sodium chloride maintains the osmotic balance. There is a compound in low concentration for detoxify metabolic by-products. The medium contains reducing and buffering agents. Selective salts inhibit the growth of molds, Enterococci and other Gram-negative bacteria. Another compound inhibits glycolysis by inactivating glyceraldehyde-3-phosphate dehydrogenase present and important in different bacteria and fungi (also Streptococci sp.). Three antibiotics are the selective agents and inhibit the accompanying bacterial flora like Bacilli, Enterobacteriaceae and Pseudomonas. Bifidobacteria can reduce an azo compound present in the medium, which gives the colonies a pink-purple coloration.

[00137] SBSM contains casein peptone (12 g/l), meat peptone (5 g/l), sodium chloride (5 g/l), beef extract (3 g/l), yeast extract (3 g/l), cornstarch (1g/l), glucose (2.5 g/l), lactulose (2.5 g/l), cysteine-hydrochloride (0.5 g/l), riboflavin (0.01 g/l), and propionic acid (99%; Smifl). All components were mixed and suspended in distilled water. The pH was adjusted to

5.5 +/- 0.2 with 5N NaOH.

[00138] All plates were incubated under suitable conditions (e.g., Table 1). Well isolated colonies were picked randomly from agar plates. Optionally, the colonies may be streaked several times on nonselective or selective media. Alternatively, isolated colonies may be pooled together and submitted again (several times) to the isolation process.

[00139] 1.2. Growth and maintenance of isolates

[00140] Well separated colonies were randomly picked from agar plates, inoculated into corresponding broth mediums and grown under suitable conditions. The resulting cultures may partly be used for production of cryo-stocks and partly used for future screening analysis. For screening analysis, bacteria cultures were centrifuged, washed with PBS and re-suspended in PBS. Alternatively, bacteria cultures were further cultivated in suboptimal medium (that does not promote culture growth) or in a medium containing propionic acid at 1 98to 10ml/l. Such bacterial cultures were than centrifuged, washed with PBS and re-suspended LU101096 | in PBS.

[00141] 1.3. Identification of isolates

[00142] Preliminary identification of isolated bacteria was based on microbiological | analysis (e.g., Gram staining and microscopic analysis) and biochemical analysis (e.g. with rapid ID 32A biochemical test kits (BioMérieux, Marcy l'Etoile, France)).

[00143] Alternatively, the characterization was performed by Bruker Biotyper (version

2.0) matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Briefly, colonies were directly picked and applied as a thin film onto a polish steel plate and allowed to dry at room temperature. Subsequently, 1ul of MALDI matrix (Bruker Daltonics) in 50% acetonitrile and 2.5% trifluoroacetic acid was applied and allowed to dry again.

[00144] For the extraction method, 1 to 2 colonies (or a few colonies in the case of a small colony size) were suspended in 300ul of molecular-grade water (Sigma-Aldrich, St. Louis, MO) and vortexed. Next, 900ul of 100% ethanol (Sigma-Aldrich) was added, vortexed, and centrifuged (13.400 x g) for 2 min. The supernatant was decanted, and the pellet was dried at room temperature. 10ul of 70% formic acid (Fluka [Sigma-Aldrich], St. Louis, MO) and 10ul of acetonitrile (Fluka) were added and thoroughly mixed by pipetting, followed by centrifugation (13,400 x g) for 2 min. One microliter of supernatant was spotted onto the 384- spot plate and allowed to dry at room temperature before the addition of 1ul of matrix. For each plate, a bacterial test standard (Bruker Daltonics) was included to calibrate the instrument and validate the run. MALDI-TOF MS was performed with the MicroFlex LT mass spectrometer (Bruker Daltonics) according to the manufacturer's suggested recommendations. Identification score criteria used were those recommended by the manufacturer: a score of 22,000 indicated species-level identification.

[00145] lil. Labelling, GM1 ELISA and Cytotoxicity assay

[00146] HA. Labeling of Cholera toxin

[00147] Horseradish peroxidase (HRP) labelling: Horseradish peroxidase (HRP) labelling were performed according to the manufacturer instructions using EZ-Link™ Plus Activated Peroxidase Kit (Thermo Fischer Scientific, Braunschweig, Germany, Catalog number in Mai 2018: 31489) Plus Activated Peroxidase Kit (Thermo Fisher Scientific Inc., Rockford, USA). Alkaline Phosphatase (AP) labelling: AP labelling were performed according to the manufacturer instructions using Lightning-Link® Alkaline Phosphatase Antibody Labeling Kit (Novus Biologicals Europe/UK, Abingdon, United Kingdom Catalog Number in May 2018: 702-0030). 99

[00148] 1.2. GM1 enzyme-linked immunosorbent assay (ELISA)

[00149] 50ul purified Monosialoganglioside (GM1, 0.5 pg/ml in phosphate-buffered saline [PBS], pH 7.2; Sigma, Hannover, Germany) were added per well of a Maxisorp ELISA plate (Nunc, Roskilde, Denmark) and adsorbed overnight at 4°C. Alternatively, 50ul purified Monosialoganglioside may be added per well of a Polysorp ELISA plate (Nunc, Roskilde, Denmark) and adsorbed overnight at 37°C. Coated wells were washed 3 times with 200i PBS + 0.02% Tween 20 (Identical washing steps were performed after each incubation step) followed by 30 minutes incubation with 200ul of PBS + 2% HSA for blocking. After one wash, 50 pl of labeled (e.g. HRP, AP) toxin at indicated concentration were added for 2 hours at room temperature or 37°C. After three final washes, 100ul of suitable Microwell Substrate were added (for HRP: TMB One Component HRP Microwell Substrate, tebu-bio GmbH, Offenbach, Germany, Catalog number in May 2018: TMBW-1000-01; for Alkaline Phosphatase: Alkaline Phosphatase Blue Microwell Substrate, Sigma Aldrich, Taufkirchen, Germany, catalog number in May 2018: AB0100), the reaction was stopped by addition suitable stop solution (for HRP: 2,5N H2SO4 ; for Alkaline phosphatase: Alkaline Phosphatase Stop Solution, Sigma Aldrich, Taufkirchen, Germany, catalog number in May 2018: A5852), and extinction measured at suitable wavelength (for HRP: 450nm/630 nm, for AP: 580 or 600 nm).

[00150] 1.3. Procedure for Testing the Cytotoxicity of a toxin suspension on human cells

[00151] 1.3.1. Cytotoxic cellular assay

[00152] Vero cells (kidney epithelial cells extracted from an African green monkey) were seeded in 96 well cell culture plates at 3x104 cells/well and cultivated for at least 4 days in standard McCoy Medium added with 10%FBS and 2mM butyrate. The presence of butyrate increasing the expression of toxin- receptor on the surface of the cells. The medium was removed and 50ul of a solution of Cholera Toxin was added to the cells and incubated for 1 hour at 37C. 50ul of McCoy Medium may than be added to each cell well and the cellular morphological phenotype monitored after 6 to 20 hours of incubation.

[00153] IV. Screening of bacterial strains capable of being loaded with GM1

[00154] IV.1 GM1-Loading Protocol

[00155] Bacterial cultures were grown in suitable media (e.g., Table 1), washed and re- suspended in PBS at OD1 to OD10 (e.g., 600nm) as mentioned above. Alternatively, bacteria were grown in suboptimal media (e.g., medium that does not allow optimal growth, growth rate, generation time) or a medium containing propionic acid (e.g., at concentration of 5Sml/L). 100

| Cells were than harvested through centrifugation and resuspended in PBS. The bacterial LU101096 suspensions were then loaded with 0.5 to 10ug/mi GM1 and incubated at 20°C to 60°C overnight or longer under agitation (e.g., Table 1). The cells were then extensively washed with PBS 0.02% Tween 20 before being resuspended in PBS.

[00156] IV.2. Procedure for testing the presence of GM1 on the surface of the loaded microorganism

[00157] Cells loaded with GM1 as mentioned above were diluted in PBS 2% HSA at OD1 to ODS (e.g., 600nm) and incubated at Room temperature (RT) or 37°C for 1 hour to prevent unspecific binding. The cells were washed once, re-suspended in PBS-1%HSA, added with the labelled (e.g., HRP or AP) CT and incubated for 1 to 2 hours at RT or 37°C.

[00158] Cells were washed 3 times with PBS + 0.02 % Tween 20 and resuspended in PBS. SOul of bacterial suspensions were given in triplicate on an ELISA plate. Two Wells were inoculated with a suitable substrate (for HRP or AP), the reactions were stopped, and extinction measured at a suitable wavelength (e.g., 580nm for AP, 450 and 630 (as reference) for HRP). The third well was used to determine the OD in the final cell suspension and the extinction measured previously and standardized to OD1. Standardized results allowed to compare the binding strength of different strains and/or preparations.

[00159] IV.3. Procedure for testing of the stability of the association between GM1 and the bacteria

[00160] To test the stability of the association between the microorganism and GM1, the GM1-loaded bacteria were subjected to different treatments before being tested for their ability to bind CT as described above. Thus, the loaded cells were subjected to pasteurization (e.g., 15 min at 70°C), to freeze and thawed, to lyophilization, to incubation at pH1 for 1 hour at 37°C, to prolonged incubation (overnight) at 37°C, to the action of gastric juice for 30 min at 37°C and storage at 4°C for 4 weeks. None of the mentioned treatment, induced a reduction of the CT-binding of the GM1-loaded Bacteria of more than 20%.

[00161] IV.4. Procedure for testing the toxin binding capacity of microorganisms loaded with GM1

[00162] Microorganisms loaded with GM1 as mentioned above were diluted in PBS 2% HSA at OD1 to ODS and incubated at Room temperature (RT) or 37°C for 1 hour to prevent any later unspecific binding. The cells were washed once, resuspended in PBS- 1%HSA and added with several concentrations (e.g., between 50ng/ml and 1mg/ml) of the natural CT and incubated for 2 hours at RT or 37°C under gently agitation. As a positive control, the same concentration of natural CT was added the PBS-1%HSA without any 101loaded microorganism. The cell suspensions were centrifuged, and the supernatants were | J101096 sterile filtered to avoid the presence of any residual cells.

[00163] The supernatants were used in the cytotoxic cellular assay as described above. Comparing the cytotoxic effect induced by the supernatant of CT incubated with loaded bacteria to the cytotoxic effect induced by the supernatant of CT not incubated with loaded bacteria may allow to quantify the toxin binding capacity of the loaded bacteria.

[00164] IV.5. Procedure for testing the in vitro toxin-neutralization capacity of microorganisms loaded with GM1

[00165] GM1 were coated on ELISA plates and the wells blocks with HSA as mentioned above. Microorganisms loaded with GM1 as described above were diluted in PBS 2% HSA at OD1 to OD5 and incubated at Room temperature (RT) or 37°C for 1 hour to prevent any later unspecific binding. GM1 loaded bacteria and labelled CT were given (at different concentrations: e.g. 1 ng/mi to 2ug/ml and OD600nm 1 to 10 for CT and GM1 loaded bacetria respectively)) at the same time on coated GM1 and incubated at RT or 37°C for 1 to 2 hours. CT without GM1 Loaded bacteria were used as positive control. The plates were washed, and the extinction measured as mentioned above. Comparing the signal obtained without GM1-loaded bacteria to the signal obtained in presence of GM1-loaded bacteria allowed to quantify the in vitro CT-neutralization capacity of the GM-loaded bacteria.

[00166] V. Results

[00167] V.1. Loading protocol

[00168] Cells (e.g., bacterial cells) were cultivated in suitable media under anaerobic conditions at a suitable growth temperature (e.g., Table 1). Cells were than harvested through centrifugation and resuspended at a suitable loading concentration, in suitable loading medium, at suitable loading pH, added with 1ug/ml GM1 and incubated at suitable loading temperature (e.g., Table 1 and procedures described above). A number of suitable growth media was identified (e.g., Table 1). For the purpose of optimal loading, the best cultivation medium was found to be not always the optimal growth medium. (e.g., Lactobacillus seems to be better loadable if previously cuitivated in BSM or SBSM, a medium in which they are barely growing, e.g., Table 1). Suitable growth temperatures were also identified (e.g., Table 1). For the purpose of optimal loading, the best cultivation temperature seems not always to be the optimal growth temperature. Temperature used were between 30 and 55°C (Table 1.). Suitable loading concentration (of the bacteria) were found to be between OD 1 and OD 10. Suitable loading pH was found to be in the range of pH from 3 to 7 with pH from 6 to 5 being optimal (e.g., Table 1). Suitable loading media was similarly 102identified (e.g., Table 1). Suitable loading temperature was also identified (e.g., Table 1). | 101096 Temperature used were between 30 and 55°C (e.g., Table 1).

[00169] Table 1. Correlation of the suitable combinations of specific microorganism, culture medium (i.e., growth medium), growth temperature, temperature of adding of said solubilized heterologous lipid carrier (i.e., loading temperature), pH and loading medium with the read-outs of the Cholera Toxin Binding assays. Strain Growth Growth | Loading Loading Loading Cholera Medium Temp Temp. pH Medium Toxin Binding Lactobacillus MRS 37°C 46°C BSM/PBS | (+) SS A rl MRS + 37°C 46°C BSM/PBS | (++) propionic (1 to 1) acid 5mi/L MRS 37°C 37°C BSM/PBS | (++) (1 to 1),

BSM OR

SBSM Lactobacillus BSM + 37°C 46°C BSM/PBS | (++) paracasei & propionic (1 to 1) Lactobacillus acid 5mi/L reuteri Lactobacillus BSM OR 37°C 46°C BSM/PBS | (++) paracasei & SBSM (1 to 1) Lactobacillus reuteri Lactobacillus BSM OR 37°C 46°C 5 BSM/PBS | (+) paracasei & SBSM (1 to 1) Lactobacillus reuteri Lactobacillus BSM OR 37°C 46°C 4 BSM/PBS | (+) paracasei & SBSM (1 to 1) Lactobacillus reuteri 103

Lactobacillus BSM OR 37°C 37°C BSM/PBS | (++) LU101096 paracasei & SBSM (1 to 1), Lactobacillus BSM ORreuteri SBSM Lactobacillus BSM OR 46°C 37°C BSM/PBS | (++) paracasei & SBSM (1 to 1) Lactobacillusreuteri

Lactobacillus BSM OR 46°C 46°C BSM/PBS | (++) paracasei & SBSM (1 to 1) Lactobacillusreuteri

Lactobacillus BSM OR 46°C 55°C BSM/PBS | (++) paracasei & SBSM (1to 1) Lactobacillusreuteri

Lactobacillus BSM OR 55°C 37°C BSM/PBS | (++) paracasei & SBSM (1 to 1) Lactobacillusreuteri

Lactobacillus BSM OR 55°C 46°C BSM/PBS | (++) paracasei & SBSM (1 to 1) Lactobacillusreuteri

Lactobacillus BSM OR 55°C 55°C BSM/PBS | (++) paracasei & SBSM (1to 1) Lactobacillusreuteri

Lactobacillus BSM OR 37°C 37°C BSM/PBS | (++) paracasei & SBSM (1 to 1) Lactobacillusreuteri

Lactobacillus BSM OR 37°C 46°C BSM/PBS | (++) paracasei & SBSM (1 to 1) Lactobacillusreuteri Tacobecilis_|BSWOR jure [se | | BSMPBS [G9 —

104paracasei & SBSM (1 to 1) LU101096 Lactobacillus =" 1 Lactobacillus BSM OR 37°C 55°C 3 PBS (+) paracasei & SBSM Lactobacillus reuteri Bifidobacterium | BSM OR 37°C 37°C 7 PBS (++) Sea Bifidobacterium | BSM OR 37°C 46°C BSM/PBS | (++) ea ||" IS Bifidobacterium | BSM OR 37°C 37°C 7 PBS (+) onan | TCO Bifidobacterium | BSM OR 37°C 46°C BSM/PBS | (++) on oll a el Bacteroides WC 37°C 37°C 7 PBS (+) | Bacteroides BSM OR 37°C 37°C 7 PBS (++) mama ol a LT TE Bacteroides WC 37°C 46°C BSM/PBS | (+)

AH Bacteroides BSM or 37°C 46°C BSM/PBS | (++)

A HN Enterococcus | MRS 37°C 46°C BSM/PBS | (+) ww | dem Enterococcus | MRS + 37°C 46°C BSM/PBS | (++) faecalis propionic (1to 1) acid 5ml/L

[00170] MRS: De Man, Rogosa and Sharpe Medium. BSM: Bifidobacterium specific Medium. SBSM: Self made Bifidobacterium Specific Medium. “(+/-)’ — weak binding, “(+)” — average binding, “(++)” — strong binding. WC Medium: Tryptone 10g/L; Gelatin peptone 10g/L; Yeast extract 5g/L; Glucose 1g/L; Sodium chloride 5g/L; L-Arginine 1g/L; Sodium pyruvate 1g/L; Menadione 0.0005g/L; Haemin 0.005 g/L (pH=7+0.2). WC Agar: Tryptone 10g/L; Gelatin peptone 10g/L; Yeast extract 5g/L; Glucose 1g/L; Sodium chloride 5g/L; L- Arginine 1g/L; Sodium pyruvate 1g/L; Menadione 0.0005g/L; Haemin 0.005 g/L; Agar 10g/L 105 EE ————

(pH=7+0.2). MRS Agar: Peptone 10 g/l; Yeast extract 4 g/l; Beef extract 8 g/l; Glucose 20 g/l; LU101096 Dipotassium phosphate 2 g/l; Sodium acetate 5 g/l; Ammonium citrate 2 g/l; Magnesium sulphate (MgSO4) 0.2 g/l; Manganese sulphate 0.05 g/l; Polysorbate 80 1 g/l; Agar 10 g/l: pH= 6.2 +0.2. BSM Agar: (Sigma Aldrich Catalog Number in May 2018: 88517). Principle and Interpretation: BSM contains Peptone and Meat extract as sources of carbon, nitrogen, vitamins and minerals. Yeast extract supplies B-complex vitamins which stimulate bacterial growth. Dextrose is the carbohydrate source. Sodium chloride maintains the osmotic balance. There is a compound in low concentration for detoxify metabolic by-products. The medium contains reducing and buffering agents. Selective salts inhibit the growth of molds, Enterococci and other Gram-negative bacteria. Another compound inhibits glycolysis by inactivating glyceraldehyde-3-phosphate dehydrogenase present and important in different bacteria and fungi (also Streptococci sp.). Three antibiotics are the selective agents and inhibit the accompanying bacterial flora like Bacilli, Enterobacteriaceae and Pseudomonas. Bifidobacteria that can reduce an azo compound present in the medium, which gives the colonies a pink-purple coloration. SBSM agar contains casein peptone (12 g/l), meat peptone (5 g/l), sodium chloride (5 g/l), beef extract (3 g/l), yeast extract (3 g/l), cornstarch (1g/l), glucose (2.5 g/l), lactulose (2.5 g/l), cysteine-hydrochiloride (0.5 g/l), riboflavin (0.01 g/l), propionic acid (99%; 5ml/l) and Agar 10g/l. All components were mixed and suspended in distilled water. The pH was adjusted to 5.5 +/- 0.2 with 5N NaOH.

[00171] BSM Broth: (Sigma Aldrich Catalog Number in May 2018: 90273). Principle and Interpretation: BSM contains Peptone and Meat extract as sources of carbon, nitrogen, vitamins and minerals. Yeast extract supplies B-complex vitamins which stimulate bacterial growth. Dextrose is the carbohydrate source. Sodium chloride maintains the osmotic balance. There is a compound in low concentration for detoxify metabolic by-products. The medium contains reducing and buffering agents. Selective salts inhibit the growth of molds, Enterococci and other Gram-negative bacteria. Another compound inhibits glycolysis by inactivating glyceraldehyde-3-phosphate dehydrogenase present and important in different bacteria and fungi (also Streptococci sp.). Three antibiotics are the selective agents and inhibit the accompanying bacterial flora like Bacilli, Enterobacteriaceae and Pseudomonas.

Bifidobacteria can reduce an azo compound present in the medium, which gives the colonies a pink-purple coloration.

[00172] SBSM contains casein peptone (12 g/l), meat peptone (5 g/l), sodium chloride (5 g/l), beef extract (3 g/l), yeast extract (3 g/l), cornstarch (1g/l), glucose (2.5 g/l), lactulose (2.5 g/l), cysteine-hydrochloride (0.5 g/l), riboflavin (0.01 g/l) and propionic acid (99%; Smif).

All components were mixed and suspended in distilled water. The pH was adjusted to 5.5 +/-

0.2 with 5N NaOH. 106 ———————

[00173] It was also found that if Lactobacillus bacteria are cultivated in BSM medium | 101096 the loading of GM1 was successful even when GM1 was added directly to the cultivating medium. For Stx2: Loading with GB3 in presence of cholesterol strongly increased the binding of Stx2. Loading in presence of cholesterol alone resulted in no bidnnig at all.

[00174] V.2. GM1 binding capacity of selected isolates

[00175] To test the ability of selected bacterial isolates to incorporate the Glycolipid GM1 and to present the oligosaccharide moiety to the environment (e.g., on the exterior cell surface), said isolates were loaded and incubated with as tested through their ability after loading to bind HRP-cholera toxin. This was tested by the means of ELISA. As presented in Figure 1, the HRP-cholera toxin bound to several isolates after those isolates have been loaded with GM1.

[00176] V.3. Characterization of the Strains

[00177] The strains designated as “BLP” were characterized as belonging to the species Lactobacillus paracasei, the strains designated as “BLR” and “BL9” to the species Lactobacillus reuteri.

[00178] V.4. Stability of the association with GM1

[00179] The stability of the association between the bacteria and GM1 was assayed for the strain BLP by means of ELISA using labelled Cholera Toxin after pre-treatment of the loaded bacteria as described in material and methods. As presented in Figure 2, none of the pre-treatment was accompanied by the loss of the bound GM1 suggesting that the binding is stable.

[00180] V.5. Loading is independent on the carbohydrate moiety of GM1

[00181] To identify which part of the GM1 molecule is involved in the interaction with our bacterial strain LBP, the strain was loaded as described in material and methods with either GM1 or GM1a (the carbohydrate part of GM1). The presence of the Carbohydrate moiety was analyzed by ELISA using HRP-CT. In contrast to the strong signal observed with bacteria loaded with GM1, no signal could be obtained with bacteria loaded with GM1a. This result demonstrates that the interaction between GM1 and the bacteria involves the sphingosine and/or fatty acid part of the GM1 molecule.

[00182] Concluding remarks

[00183] The present invention is the first one to report on non-pathogenic microorganisms naturally and stably interacting with the sphingolipid part of the 107

Monosialoganglioside GM1 in a way that the carbohydrate part of the GM1 stays available to LU101096 the environment, thereby allowing the binding of the Cholera Toxin. A number of bacterial strains presenting a binding of the GM1 were isolated out of the human gut (e.g., Table 1). The binding was dependent on the sphingolipid part of the GM1 structure and was resistant to several treatments.

[00184] These surprising results unequivocally demonstrate that the present invention allows to produce non-GMO and non-pathogenic microorganisms that present a receptor for the cholera toxin on their surface. Such microorganisms may be further developed into medicaments or food supplements to be administered orally to human subjects or animal specimen in viable or killed form to abate, cure, treat or prevent diseases related with Cholera Toxin (or other toxins).

[00185] One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. Further, it will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention. The compositions, methods, procedures, treatments, molecules and specific compounds described herein are presently representative of certain embodiments are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art which are encompassed within the spirit of the invention are defined by the scope of the claims.

[00186] The invention illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising”, “including,” containing”, etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by exemplary embodiments and optional features, modification and variation of the inventions embodied herein may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention.

[00187] The invention has been described broadly and generically herein. Each of the narrower species and sub-generic groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not 108the excised material is specifically recited herein.

Other embodiments are within the following LU101096 claims. 109

Claims (5)

WHAT IS CLAIMED IS

1. A modified microorganism comprising: i) a cell and ii) a heterologous lipid carrier, said lipid carrier comprising: | a) a lipid portion, wherein said lipid portion is at least partially associated with an exterior surface of said cell of said modified microorganism, wherein said lipid portion comprising: a ceramide-like glycolipid moiety and/or a fatty acid moiety; preferably said exterior surface of said cell comprising: a cell wall and/or a cell membrane and/or an outer cell membrane and/or a polysaccharide (e.g., capsule polysaccharide); further preferably said lipid portion is at least partially incorporated and/or adhered and/or bound to said exterior surface of said cell; and b) a non-lipid portion, wherein said microorganism is capable of locating and/or displaying said non-lipid portion or fragment thereof onto the exterior surface of said cell, preferably said non-lipid portion comprising a carbohydrate moiety and/or lipopeptide moiety (e.g., a glycosylated lipopeptide moiety, e.g., a carbohydrate antigen); iii) optionally, said modified microorganism (e.g., said lipid carrier) further comprising a heterologous steroid moiety; preferably said steroid moiety is cholesterol, a derivative or analogue thereof.

2. The modified microorganism according to any one of preceding claims, wherein said non-lipid portion is capable of one or more of the following characteristics: i) binding and/or reducing toxicity of and/or neutralizing a toxin; i) binding and/or reducing the pathogenicity and/or neutralizing a pathogenic microorganism (e.g., a bacterium, virus, fungus or protozoa), iii) binding a receptor of a toxin; and | | iv) binding a receptor of a pathogenic microorganism.

3. The modified microorganism according to any one of preceding claims, wherein said modified microorganism has one or more of the following characteristics: 110i) said cell of said microorganism comprising a cell membrane (e.g., yeast or Lu101096 protozoa); preferably said cell membrane further comprising a peptidoglycan; ii) said cell of said microorganism comprising a cell wall, preferably said cell wall comprising a peptidoglycan or a mannoprotein; | 5 iii) said cell of said microorganism comprising a cell membrane, wherein said cell membrane further comprising a peptidoglycan; iv) said cell of said microorganism is not comprising an outer membrane, preferably said microorganism is a gram-positive bacterium.

Vv) said cell of said microorganism comprising an outer membrane; preferably said microorganism is a gram-negative bacterium; vi) said cell of said microorganism is not comprising a recombinant and/or fusion polypeptide, preferably said recombinant and/or fusion polypeptide is obtained by the means of artificial genetic manipulation; vii) said microorganism is non-pathogenic.

4 The modified microorganism according to any one of preceding claims, wherein said lipid carrier has one or more of the following characteristics: i) comprising a glyceride moiety; preferably said glyceride moiety comprises at least one fatty acid, further preferably said lipid portion of said lipid carrier comprises said glyceride moiety; ii) comprising a ceramide moiety, wherein preferably: a) said ceramide moiety is composed of an amino alcohol (e.g., sphingosine) and/or a fatty acid, further preferably said lipid portion of said lipid carrier comprises said ceramide moiety; and/or ii) comprising a carbohydrate moiety; preferably said non-lipid portion of said lipid carrier comprises said carbohydrate moiety; further preferably said carbohydrate moiety is B- or a- linked to said ceramide moiety (e.g., as in item (ii)), further preferably said carbohydrate is not a monosaccharide or a disaccharide moiety; further preferably said carbohydrate is selected from the group consisting of: an oligosaccharide and a polysaccharide, most preferably the first sugar of the said carbohydrate is galactose, a glucose, a mannose, a xylose, a neuraminic acid, a N-acetyl glucosamine, N-acetyl galactosamine or a galacturonic acid; 111iv) comprising one or more polypeptides (e.g., a heterologous recombinant or fusion LU101096 polypeptide, e.g., a glycosylated polypeptide or an immunologically active polypeptide); preferably said non-lipid portion comprising said one or more polypeptides; further preferably said one or more polypeptides is an enzyme, a cytokine or a chemokine, a peptidomimetic compound, an antigen, an antibody (e.g., a single chain or a single domain antibody), a fragment or derivative thereof; v) comprising a pharmaceutically active compound; preferably said non-lipid portion comprising said pharmaceutically active compound; vi) is not comprising a recombinant and/or fusion polypeptide (e.g., an endogenous recombinant and/or fusion polypeptide), preferably said recombinant and/or fusion polypeptide is obtained by the means of artificial genetic manipulation; vii) is not expressed or synthetized by said microorganism; viii)is at least partially expressed or synthetized by said microorganism (e.g., endogenously synthetized), preferably said microorganism is capable of expressing or synthetizing a ceramide or sphingolipid moiety (e.g., a Sphingomonas sp. Gram-negative bacterium); ix) said lipid portion is not comprising polypeptides; X) is not comprising a transmembrane polypeptide or a polypeptide membrane anchor domain; xi) is not susceptible to proteolysis (e.g., under physiological conditions); xii) is not immunogenic to a mammalian host, preferably said mammalian host is human; xiii)is immunogenic to a mammalian host, preferably said mammalian host is human; xiv)is not covalently bound to said cell membrane of said cell of said modified microorganism; xv) comprising a glycolipid; xvi)comprising a lipopeptide (e.g., a glycosylated lipopeptide).

5. The modified microorganism according to any one of preceding claims, wherein said lipid carrier is selected from the group consisting of: i) Monosialotetrahexosylganglioside (GM1), ii) Globotriaosylceramide (Gb3), 112iii) Ganglioside GD1a, LU101096 iv) Gangliosides GM2, GD2, GD1b, GT1b, GT1c, GQ1c, GA1, GM1b, v) Gangliosides GM3, GD3 und GT3, vi) Gangliosides Gb4, Blood Group Type |, Type 2, Blood Group A, Blood Group B, Blood Group H, Blood Group H Type 1, Blood Group H Type 2, Blood Group H Type 3, Lewis y, Lewis a, Lewis b, Lewis x, H, Sialyl Lewis x, Sialyl Lewis a, Sialyl | Lewis b, Sialyl Lewis x, alpha Gal epitope, Gal a1-3Gal1-4GlacNAc, Gal(a 1- 4)Gal(R1-4)GIcNAc-R, Gal(a 1-4)Gal(31-4)Glc NAc-(R1-3)Gal(31-4)Glc-R, Gal(a 1-4)Gal(R1-4)GIcNAc(B1-2) Man-R vii) any one of the foilowing: 2-6 Sialyl i- | Neu5Ac(a 2-6)Gal(R1-4)GIcNAc(B1-3)Gal(81-4)[Fuc(a 1-3)]GIcNAc(B1- Lewis x )-R 3'-Sulfo Lewis a HSO3(-3)Gal(R1-3)[Fuc(a1-4)]GIcNAc(B1-)-R 3'-Sulfo Lewis x HSO3(-3)Gal(B1-4)[Fuc(a1-3)]GIcNAc(R1-)-R 6,6'- Disulfo Sialyl Lewis | Neu5Ac(a 2-3)[HSO3(-6)]Gal(81-4)[Fuc(a 1-3)][HSO3(-6)]GIcNAc(B1-)- x R 6-Suifo Lewis x Gal(R1-4)[Fuc(a 1-3)][HSO3(-6)]GIcNAc(B1-)-R 6'-Sulfo Sialyl Lewis x Neu5Ac(a 2-3)[HSO3(-6)]Gal(R1-4)[Fuc(a 1-3)]GIcNAc(B1-}R gléleNAe)_su- SLe* Neu5Aca2-3GalB1-4(Fuca1-3)(6-O-Su)GIcNAcB-R 6'-Sia-6- | Su-LacNAc | Neu5Aca2-6GalB1-4(6-O-Su)GIcNAcB-R | Neu5Aca2-3GalB1-3(6-0-Su)GIcNAcB-R 113

3'Siale® LU101096 6-Su- 3 SLN Neu5Aca2-3GalB1-4(6-O-Su)GIcNAcCB-R 3'SLN(Gc) | Neu5Gca2-3GalB1-4GIcNAcB-R 6'SLN(Gc) | Neu5Gca2-6GalB1-4GIcNAcB-R GIcNAcB3" GIcNAcB1-3GalB1-4GIcNAcB-R LacNAc Isomaltotrio Glca1-6Glca1-6GIcB-R se GIcNAcB1-4GIcNAB1-4GIcNAcB-R alpha Gal epitope, Gal(a 1-3)Gal(R1-4)GIcNAc-R GIcNAcB1,3ManB1,4Glcf - R GalNAca1-3(Fuca1-2)GalB-R Gala1-3(Fuca1-2)GalB-R Gal3,4- GlcNAc GalB1-4(GalB1-3)GIcNAcB-R asialo- DGalp(R1-3)DGalpNAc(B1-4)DGalp(B1-4)DGlcp(B1-1)-R a - a c(R1- a - c -1)- GM1,GA1 P P P P asialo- DGalpNAc(RB1-4)DGalp(R1-4)DGlcp(R1-1)-R a c(R1- a - e -1)- GM2,GA2 P P P Blood Group À Trisacchari de GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-)-R Blood Group À Type 1 GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-3)GICNAc(B1-)-R Blood GalNAc(a 1-3)[Fuc(a 1-2)]Gal(R1-3)[Fuc(a 1-4)}GIcNAc(B1-)-R Group À | 114