NZ760637B2 - Compositions comprising bacterial strains - Google Patents

Abstract

The invention provides compositions comprising bacterial strains, in particular Parabacteroides distasonis (strain NCIMB 42382,) for treating and preventing a neurodegenerative disorder or brain injury. Neurodegenerative disorders to be treated include Parkinson’s disease, including progressive supranuclear palsy, progressive supranuclear palsy, Steele-Richardson-Olszewski syndrome, normal pressure hydrocephalus, vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism; Alzheimer’s disease, including Benson's syndrome; Huntington’s disease; amyotrophic lateral sclerosis; Lou Gehrig's disease; motor neurone disease; prion disease; spinocerebellar ataxia; spinal muscular atrophy; dementia, including Lewy body, vascular and frontotemporal dementia; primary progressive aphasia; mild cognitive impairment; HIV-related cognitive impairment and corticobasal degeneration. Brain injuries to be treated include stroke, such as cerebral ischemia, focal cerebral ischemia, ischemic stroke or hemorrhagic stroke. anuclear palsy, progressive supranuclear palsy, Steele-Richardson-Olszewski syndrome, normal pressure hydrocephalus, vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism; Alzheimer’s disease, including Benson's syndrome; Huntington’s disease; amyotrophic lateral sclerosis; Lou Gehrig's disease; motor neurone disease; prion disease; spinocerebellar ataxia; spinal muscular atrophy; dementia, including Lewy body, vascular and frontotemporal dementia; primary progressive aphasia; mild cognitive impairment; HIV-related cognitive impairment and corticobasal degeneration. Brain injuries to be treated include stroke, such as cerebral ischemia, focal cerebral ischemia, ischemic stroke or hemorrhagic stroke.

Description

COMPOSITIONS COMPRISING IAL STRAINS TECHNICAL FIELD This invention is in the field of compositions comprising bacterial strains isolated from the mammalian digestive tract and the use of such compositions in the treatment of disease.

BACKGROUND TO THE INVENTION The human intestine is thought to be sterile in utero, but it is exposed to a large variety of maternal and environmental microbes immediately after birth. Thereafter, a dynamic period of microbial zation and sion occurs, which is influenced by factors such as ry mode, environment, diet and host genotype, all of which impact upon the composition of the gut iota, particularly during early life. Subsequently, the microbiota stabilizes and becomes adult-like [1]. The human gut microbiota contains more than 500-1000 different phylotypes belonging essentially to two major ial ons, the Bacteroidetes and the Firmicutes [2]. The successful symbiotic relationships arising from bacterial colonization of the human gut have yielded a wide variety of metabolic, structural, protective and other beneficial functions. The enhanced metabolic activities of the colonized gut ensure that otherwise indigestible dietary components are degraded with e of by—products providing an important nutrient source for the host. Similarly, the immunological importance of the gut microbiota is ecognized and is exemplified in germfree animals which have an impaired immune system that is functionally reconstituted ing the introduction of sal bacteria [3-5].

Dramatic changes in microbiota composition have been documented in gastrointestinal disorders such as inflammatory bowel disease (IBD). For example, the levels of Clostridium cluster XIVa bacteria are reduced in IBD patients whilst numbers of E. coli are sed, suggesting a shift in the balance of symbionts and ionts within the gut [6-9].

In recognition of the potential positive effect that certain bacterial strains may have on the animal gut, various s have been proposed for use in the treatment of various diseases (see, for example, [10— 13]). Also, certain strains, ing mostly Lactobacillus and Bifidobacterium strains, have been proposed for use in treating various inflammatory and autoimmune diseases that are not ly linked to the intestines (see [14] and [15] for reviews). However, the relationship between ent diseases and different bacterial strains, and the precise effects of particular bacterial s on the gut and at a systemic level and on any particular types of diseases are poorly characterised, particularly for neurodegenerative disorders.

Recently, there has been increased interest in the art regarding alterations in the gut microbiome that may play a pathophysiological role in human brain diseases [16]. Preclinical and clinical evidence are strongly ting a link between brain development and microbiota [17]. A growing body of preclinical literature has demonstrated bidirectional signalling between the brain and the gut microbiome, involving multiple neurocrine and ine signalling systems. Indeed, increased levels [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm ionNone set by kjm [Annotation] kjm Unmarked set by kjm of Clostridium species in the microbiome have been linked to brain disorders [18], and an imbalance of the Bacteroidetes and Firmicutes phyla has also been ated in brain development disorders . tions that altered levels of gut commensals, including those of Bifidobacterz’um, Lactobacillus, Sutterella, ella and Ruminococcus genera and of the Alcaligenaceae family are involved in immune-mediated central nervous system (CNS) disorders, are oned by studies suggesting a lack of alteration in the microbiota between patients and healthy subjects [10].

Parabacteroides distasonis has been proposed for treating a variety of disorders including asthma, rheumatoid arthritis and multiple sclerosis [20] Like asthma and toid arthritis, multiple sclerosis is primarily mediated by the immune system.

The immune system attacks ated axons in the central nervous system, destroying the myelin called s or lesions. ination occurs in ular in the optic nerves, subpial spinal cord, brainstem, cerebellum, and juxtacortical and periventricular white matter regions.

As such, multiple sclerosis has a different pathology to other neurodegenerative diseases, such as Parkinson's disease, Alzheimer’s disease or dementia. For example, multiple sclerosis is commonly diagnosed in ts in their 20s and 30s, while many other neurodegenerative diseases, such as Parkinson's disease, Alzheimer’s and dementia, are sed predominantly in patients aged over 65 years old.

Parkinson's disease, like many neurodegenerative diseases, is primarily mediated by the accumulation of misfolded protein. Parkinson's disease is a einopathology that involves the accumulation of or-synuclein, which aggregate as insoluble fibrils in Lewy bodies within the cytoplasm of the neuronal body. The accumulation of clein is toxic and impairs the functions of mitochondria, mes, and endoplasmic reticulum, and interferes with microtubule transport.

Nonsteroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen, have been tested for their efficacy in treating a variety of neurological diseases, but the clinical impact ofNSAIDs on neurodegenerative diseases like son’s disease remains unclear. While some studies showed that chronic NSAID use is protective against Parkinson’s disease, other studies could not confirm the existence of a significant onship. A recent meta-analysis indicated that the use of non-aspirin NSAID, particularly fen, reduces the risk of PD by 15% while the use of aspirin did not show any effect [2 1 ].

At present, the practical effect of the link between the microbiome and human brain diseases is poorly characterised. Accordingly, more direct analytical studies are required to identify the therapeutic impact of altering the microbiome on neurodegenerative disorders.

There is a requirement in the art for new methods of treating neurodegenerative disorders. There is also ent for the potential effects of gut bacteria to be characterised so that new ies using acteria can be developed.

SUMMARY OF THE INVENTION The inventors have developed new therapies for treating and preventing neurodegenerative disorders.

Accordingly, in one aspect, the present invention provides the use of a composition comprising a bacterial strain of the genus cteroides , in the manufacture of a medicament for ng or preventing a neurodegenerative er selected from the group consisting of Parkinson’s disease, including progressive supranuclear palsy, progressive supranuclear palsy, - Richardson-Olszewski syndrome, normal pressure hydrocephalus, vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism; Alzheimer’s disease, including Benson's syndrome; Huntington’s disease; amyotrophic lateral sclerosis; Lou Gehrig's disease; motor e disease; prion disease; spinocerebellar ataxia; spinal ar atrophy; dementia, including Lewy body, vascular and temporal dementia; y progressive aphasia; mild ive ment; HIV-related cognitive impairment and obasal degeneration in a patient.

In another aspect, the present invention provides the use of a composition comprising a bacterial strain of the genus Parabacteroides , in the manufacture of a medicament for treating brain injury in a patient.

In another aspect, the present invention provides a food product comprising the composition comprising a ial strain of the genus Parabacteroides , when used in a method of the invention.

In another aspect, the present invention provides the use of a food product sing the composition comprising a bacterial strain of the genus Parabacteroides , in the manufacture of a medicament for treating or preventing a neurodegenerative disorder in a patient, n the neurodegenerative disorder is not multiple sclerosis.

The inventors have identified that bacterial strains from the genus Parabacteroides may be effective for ng neurodegenerative diseases. As described in the examples, administration of compositions comprising Parabacteroides distasonis can protect against reactive oxygen species and prevent inflammation, thus acting as a neuroprotectant. The inventors have also identified that treatment with Parabacteroides distasonis can reduce the activation of proinflammatory molecules, such as NFκB and IL-6, by LPS and mutant α-synuclein A53T. The inventors have identified that treatment with cteroides distasonis can reduce histone deacetylation activity and lipid peroxidation in vitro, which can help to reduce cell death and apoptosis. The ors have also identified that Parabacteroides distasonis can produce indole that can attenuate inflammation and oxidative stress. Furthermore, the inventors have demonstrated that treatment with Parabacteroides distasonis can increase kynurenine levels.

The inventors have also found that ent with Parabacteroides distasonis increases the activation of BDNF. BDNF is a neurotrophic growth factor that has been shown to enhance neuron differentiation and survival. Thus, the compositions described herein can be used in a method of enhancing nerve cell survival in the treatment or prevention of neurodegenerative diseases.

Disclosed herein is a composition comprising a bacterial strain of the genus Parabacteroides , for use in a method of treating or preventing a egenerative disorder.

Also disclosed herein is a composition comprising a bacterial strain of the genus Parabacteroides , for use in a method of treating or preventing a disease or ion selected from the group consisting of: Parkinson’s disease, including progressive uclear palsy, progressive supranuclear palsy, Steele-Richardson-Olszewski syndrome, normal re hydrocephalus, vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism; Alzheimer’s disease, including Benson's syndrome; multiple sclerosis; Huntington’s disease; amyotrophic lateral sclerosis; Lou Gehrig's disease; motor neurone disease; prion disease; spinocerebellar ataxia; spinal muscular atrophy; dementia, ing Lewy body, vascular and frontotemporal dementia; y progressive a; mild cognitive impairment; HIV-related ive impairment and corticobasal degeneration.

Also sed herein is a composition comprising a bacterial strain of the genus Parabacteroides , for use in a method of treating or preventing Parkinson’s e, such as environmental, familial or Parkinson’s associated with general inflammatory status. The inventors have identified that treatment with Parabacteroides strains can reduce the activation of lammatory les, such as NFκB and IL-6, by LPS and mutant α-synuclein A53T in in vitro models of environmental and familial Parkinson’s. Also sed herein is a composition comprising a bacterial strain of the species Parabacteroides distasonis, for use in the treatment of Parkinson’s e. Compositions using Parabacteroides distasonis may be particularly effective for treating Parkinson’s.

Also sed herein are compositions for use in a method of treating or preventing early-onset neurodegenerative e. In some embodiments, the compositions of the invention are for use in a method of preventing or delaying onset or progression of a neurodegenerative disorder.

In preferred embodiments, the bacterial strain in the composition disclosed herein is of Parabacteroides onis. Closely related strains may also be used, such as bacterial strains that have a 16S rRNA ce that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to the 16S rRNA sequence of a bacterial strain of Parabacteroides onis. Preferably, the bacterial strain has a 16S rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 8 or 9. Preferably, the sequence identity is to SEQ ID NO:9. Preferably, the bacterial strain disclosed herein has the 16S rRNA sequence represented by SEQ ID NO:9.

In certain ments, the ition of the invention is for oral administration. Oral stration of the strains of disclosed herein can be effective for neurodegenerative disorders.

Also, oral administration is convenient for patients and practitioners and allows delivery to and / or partial or total colonisation of the intestine.

In certain embodiments, the composition of the invention comprises one or more pharmaceutically acceptable excipients or carriers.

In certain embodiments, the composition of the invention comprises a bacterial strain that has been lyophilised. Lyophilisation is an effective and convenient technique for preparing stable compositions that allow delivery of bacteria.

In certain embodiments, the disclosure provides a food product comprising the composition as described above.

In n embodiments, the disclosure provides a vaccine composition comprising the composition as described above.

Additionally, disclosed herein is a method of treating or preventing neurodegenerative disorders, comprising administering a composition comprising a bacterial strain of the genus Parabacteroides . 4B followed by page 5 In certain embodiments, the composition disclosed herein may be for use in treating brain injury.

The neuroprotective activity of the compositions of the invention and their ability to reduce levels of histone deacetylase ty (HDAC) may make them useful for treating brain injury. In preferred embodiments, the compositions of sed herein may be for use in treating stroke, such as treating brain injury resulting from a stroke.

[Annotation] kjm None set by kjm [Annotation] kjm ionNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm BRIEF DESCRIPTION OF DRAWINGS Figure 1: Cell viability of lastoma cells Figure 2A: Downregulation of IL-6 secretion Figure 2B: gulation of IL-8 secretion Figure 3: Inhibition of oc- synuclein IL-6 and IL-8 secretion Figure 4: tion of oc- synuclein induced NFKB promoter activation Figure 5: Inhibition of LPS induced NFKB promoter activation Figure 6: Change in antioxidant capacity Figure 7: Change in total anti-oxidant capacity (lipid oxidation) Figure 8: Change in histone deacetylatase (HDAC) activity Figure 9: Level of Indole production Figure 10: Level of Kyrunenine production Figure 11: Neuroprotection — cell viability. Figure 11 shows the same data as Figure 1.

Figure 12: Levels of metabolite production — neurotransmitters in the brain Figure 13: Levels of metabolite production — organic acids in the supernatant Figure 14: Effect on intestinal r function Figure 15: Level of BDNF production Figure 16: Change in ROS levels in (A) U373 cells, (B) SH-SY5Y cells Figure 17: Production of neurotransmitters in the brain Figure 18: Changes in Hippocampal Receptor Expression — A) Oxytocin Receptor, B) Vasopressin or, C) orticoid Receptor and D) Mineralocorticoid Receptor Figure 19: Changes in Hippocampal Expression of A) Corticotropin-Releasing Hormone (CRH), B) BDNF Expression and C) TLR4 Figure 20: A) Changes in Hippocampal Corticotropin Releasing Hormone Receptor 1 ) Expression and B) Corticotropin Releasing Hormone Receptor 2 (CRFR2) Expression Figure 21: Changes in Hippocampal Expression of A) Tumour Necrosis Factor, B) Interleukin lb and C) IL-6 Figure 22: A) Changes in ampal Integrin Alpha M (CD1 lb) Expression and B) Changes in Hippoupal Serotonin lA Receptor (5-HT1A or) Expression [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Figure 23: A) Changes in ampal Glutamate Ionotropic Receptor NMDA Type Subunit 2A (Grin2A) and B) Glutamate Ionotropic Receptor NMDA Type t 2B (Grin2B) sion Figure 24: Changes in Hippocampal Expression of A) Aminobutyric Acid A Receptor 2 (GABA A2), B) Gamma-Aminobutyric Acid B Receptor 1 (GABA BRl) and C) Dopamine or 1 (DRDl) Figure 25: Changes in la Receptor Expression — A) Oxytocin Receptor, B) Vasopressin Receptor, C) Glucocorticoid Receptor and D) Mineralocorticoid Receptor Figure 26: Changes in Amygdala Expression of A) Brain Derived Neurotrophic Factor , B) Toll-like Receptor 4 (TLR-4), C) Corticotropin Releasing Hormone Receptor 1 (CRFRl) and D) Corticotropin Releasing Hormone or 2 (CRFR2) Figure 27: Changes in Amygdala Expression ofA) Integrin Alpha M (CD1 1b), B) Interleukin-6 (IL- 6), C) Glutamate Ionotropic Receptor NMDA Type Subunit 2A (Grin2A) and D) Glutamate Ionotropic Receptor NMDA Type Subunit 2B (Grin2B) Figure 28: Changes in la Expression of A) GABA-A Receptor Alpha 2 Subunit 2), B) GABA-A Type B Receptor 1 t (GABBRl) and C) Dopamine Receptor 1 (DRDl) Figure 29: Changes in Prefrontal Cortex Expression of A) Oxytocin Receptor, B) Brain Derived Neurotrophic Factor (BDNF), C) Mineralocorticoid Receptor and D) Glucocorticoid Receptor Figure 30: Changes in Prefrontal Cortex Expression of A) Toll-like Receptor 4 (TLR-4), B) Corticotropin Releasing Hormone Receptor 1 (CRFRl), C) Corticotropin Releasing Hormone or 2 (CRFR2) and Integrin Alpha M (CD1 1b) Figure 31: Changes in Prefrontal Cortex Expression of A) Interleukin-6 (IL-6), B) Glutamate Ionotropic Receptor NMDA Type Subunit 2A (Grin2A), C) Glutamate Ionotropic Receptor NMDA Type Subunit 2B (Grin2B) and D) GABA-A Receptor Alpha 2 Subunit (GABRA2) Figure 32: Changes in Prefrontal Cortex Expression of A) GABA-A or Type B Receptor Subunit 1 (GABBRl) and B) Dopamine Receptor 1 (DRDl) Figure 33: s in Colon Expression of A) Tryptophan Hydroxylase-l (Tphl) and B) Indoleamine2,3-Dioxygenase-l (IDOl) Figure 34: Changes in Ileum Expression of A) Tryptophan Hydroxylase-l (Tphl) and B) Indoleamine2,3-Dioxygenase-1 (IDOl) Figure 35: Changes in ating Tryptophan Metabolite Levels A) Kynurenine, B) Tryptophan and C) Kynurenine/ Tryptophan Index of metabolism [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm ation] kjm Unmarked set by kjm Figure 36: Effect on Interferon-y Production from mouse Splenocytes from mice fed with MRxOOOS and MRx0029 Figure 37: Effect on Interleukin-113 tion from Splenocytes Figure 38: Effect on Interleukin-6 Production from Splenocytes Figure 39: Effect on Tumour Necrosis Factor Production from Splenocytes Figure 40: Effect on Interleukin-10 Production from Splenocytes Figure 41: Effect on Chemoattractant CXCLl Production from Splenocytes Figure 42: Changes in Caecal Short Chain Fatty Acid Levels Figure 43: MRx0029 and MRXOOS-induced changes in gene expression levels of Actin, Villin, Occludin TJPl, TJP2, MAP2, DRD2, GABRB3, SYP, PINK], PARK7 and NSE.

Figure 44: SHSYSY cell differentiation induced by MRx0005 and MRx0029. (A-C) Representative images of immuno labelled cells with Phalloidin and MAP2. (D—F) images of A-C merged with DAPI images. (G—I) B3 tubulin immunolabelled cells. (J-L) merged with DAPI images.

Magnification x630. J-K) Western blot analysis of effects of 5 and MRx0029 treatment on SHSYSY cells. Western blot membranes were probed with antibodies to MAP2 (M) and b3 tubulin (N). Actin was used as a loading control. Lower panels: representative blots from one of six separate experiments; upper : ve densitometric intensity.

DISCLOSURE OF THE INVENTION Bacterial strains The itions of the invention comprise a ial strain of the genus Parabacteroia’es. The examples demonstrate that bacteria of this genus are useful for treating or preventing egenerative ers. The preferred bacterial strains are of the species Parabacteroides distasonis.

Examples of Parabacteroides species for use in the ion include Parabacteroides distasonis, cteroides goldsteinii, Parabacteroides merdae and Parabacteroides johnsonii. The Parabacteroides resemble the Bacteroides and are Gram-negative, obligately anaerobic, non-spore- forming, non-motile and rod-shaped, and 0.8—1.6><1.2—12um in size. Parabacteroides distasonis is one of the most common species in human faeces. The type strain of P. distasom's is JCM 5825T (=CCUG 4941T=DSM 20701T=ATCC 8503T) The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences ofP. distasom's strains JCM 5825T, JCM 13400, JCM 13401, JCM 13402, JCM 13403 and JCM 13404 and P. merdae strains JCM 9497T and JCM 13405 are AB23 8922—AB23 8929, respchy osed herein as SEQ ID 8). Exemplary strains are also described in [22].

[Annotation] kjm None set by kjm [Annotation] kjm ionNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm ation] kjm Unmarked set by kjm The Parabacteroides distasom's bacterium deposited under accession number NCIMB 42382 was tested in the Examples and is also referred to herein as strain 755 or MRx0005. A 16S rRNA sequence for the 755 strain that was tested is ed in SEQ ID N029. Strain 755 was ted with the international depositary authority NCIMB, Ltd. (Ferguson ng, Aberdeen, AB21 9YA, Scotland) by GT Biologics Ltd. (Life Sciences Innovation Building, Aberdeen, AB25 2ZS, nd) on 12th March 2015 as "Parabacteroides sp 755" and was assigned accession number NCIMB 42382. GT Biologics Ltd. Subsequently changed its name to 4D Pharma Research Limited. describes administration of strain 755 to mice and shows that it can affect disease processes e of the gut (such as asthma and arthritis). Strain 755 also affects disease processes outside of the gut in the treatment of neurodegenerative disorders described herein.

A genome sequence for strain 755 is provided in SEQ ID NO: 10. This sequence was generated using the PacBio RS II platform.

Bacterial strains closely related to the strain tested in the es are also ed to be effective for treating or preventing neurodegenerative disorders. In certain embodiments, the bacterial strain for use in the invention has a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to the 16s rRNA sequence of a bacterial strain of Parabaclemz‘des distasonis. Preferably, the ial strain for use in the invention has a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 8 or 9. Preferably, the sequence identity is to SEQ ID NO:9. Preferably, the bacterial strain for use in the invention has the 165 rRNA sequence represented by SEQ ID N029.

Bacterial s that are biotypes of the bacterium ted under accession number 42382 are also expected to be ive for treating or preventing neurodegenerative disorders. A biotype is a closely related strain that has the same or very similar physiological and biochemical characteristics.

Strains that are biotypes of the bacterium deposited under accession number NCIMB 423 82 and that are suitable for use in the invention may be identified by sequencing other nucleotide sequences for the bacterium deposited under accession number NCIMB 42382. For example, substantially the whole genome may be sequenced and a biotype strain for use in the ion may have at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% sequence ty across at least 80% of its whole genome (e.g. across at least 85%, 90%, 95% or 99%, or across its whole genome). Other suitable sequences for use in identifying biotype strains may include hsp60 or repetitive sequences such as BOX, ERIC, (GTG)5, or REP or [23]. Biotype strains may have sequences with at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% sequence ty to the corresponding sequence of the bacterium deposited under accession number NCIMB 42382.

In ce ' identib embodiments, the bacterial strain for use in the invention has a genome with sequence 35 SEQ ID NO: 10. In preferred embodiments, the bacterial strain for use in the invention has a genome with at least 90% sequence identity (e.g. at least 92%, 94%, 95%, 96%, 97%, 98%, 99% or [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm ation] kjm Unmarked set by kjm 100% ce identity) to SEQ ID NO:10 across at least 60% (e.g. at least 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, 99% or 100%) of SEQ ID NO:10. For example, the bacterial strain for use in the invention may have a genome with at least 90% sequence ty to SEQ ID NO: 10 across 70% of SEQ ID NO:10, or at least 90% sequence identity to SEQ ID NO:10 across 80% of SEQ ID NO:10, or at least 90% ce identity to SEQ ID NO: 10 across 90% of SEQ ID NO: 10, or at least 90% sequence ty to SEQ ID NO:10 across 100% of SEQ ID NO:10, or at least 95% sequence identity to SEQ ID NO: 10 across 70% of SEQ ID NO:10, or at least 95% sequence identity to SEQ ID NO:10 across 80% of SEQ ID NO: 10, or at least 95% sequence identity to SEQ ID NO: 10 across 90% of SEQ ID NO: 10, or at least 95% sequence identity to SEQ ID NO: 10 across 100% of SEQ ID NO:10, or at least 98% ce identity to SEQ ID NO:10 across 70% of SEQ ID NO:10, or at least 98% sequence identity to SEQ ID NO: 10 across 80% of SEQ ID NO:10, or at least 98% sequence identity to SEQ ID NO: 10 across 90% of SEQ ID NO: 10, or at least 98% ce identity to SEQ ID NO: 10 across 100% of SEQ ID NO:10.

Alternatively, strains that are biotypes of the bacterium deposited under accession number NCIMB 423 82 and that are suitable for use in the invention may be identified by using the accession number NCIMB 42382 deposit and restriction fragment analysis and/or PCR analysis, for example by using fluorescent amplified fragment length polymorphism (FAFLP) and repetitive DNA t PCR fingerprinting, or protein profiling, or l 16S or 23S rDNA sequencing. In red embodiments, such ques may be used to identify other Parabacteroides distasonis strains.

In certain embodiments, strains that are biotypes of the bacterium deposited under accession number NCIMB 42382 and that are suitable for use in the invention are strains that provide the same pattern as the bacterium deposited under accession number NCIMB 42382 when analysed by amplified ribosomal DNA restriction analysis (ARDRA), for example when using Sau3AI restriction enzyme (for exemplary s and guidance see, for example [24]). Alternatively, biotype strains are identified as strains that have the same carbohydrate fermentation patterns as the bacterium deposited under accession number NCIMB 42382.

Other Parabacteroides strains that are useful in the compositions and methods of the invention, such as biotypes of the bacteria deposited under accession number NCIMB 423 82, may be identified using any riate method or strategy, including the assays described in the examples. For instance, strains for use in the invention may be identified by culturing with neuroblastoma cells and then assessing cytokine levels and levels of neuroprotection or neuro-proliferation. In particular, bacterial strains that have similar growth patterns, metabolic type and/or surface antigens to the bacterium ted under accession number NCIMB 423 82 may be useful in the invention. A useful strain will have comparable immune modulatory activity to the NCIMB 42382 strain. In particular, a e strain will elicit able effects on the neurodegenerative disease models and comparable effects on cytDe levels to the effects shown in the Examples, which may be identified by using the culturing and administration protocols described in the Examples. ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm A particularly preferred strain of the invention is the Parabacteroides distasonis strain deposited under accession number NCIMB 423 82. This is the exemplary 755 strain tested in the examples and shown to be effective for treating disease. Therefore, the ion provides a cell, such as an isolated cell, of the Parabacteroides distasonis strain deposited under accession number NCIMB 423 82, or a derivative thereof. The invention also provides a composition comprising a cell of the Parabactemides distasonz's strain deposited under accession number NCIMB 42382, or a derivative thereof. The invention also provides a ically pure culture ofthe Parabacteroides distasom's strain deposited under accession number NCIMB 42382. The ion also provides a cell of the Parabacteroides distasonis strain deposited under accession number NCIMB 42382, or a derivative thereof, for use in therapy, in particular for the diseases described herein.

A derivative of the strain deposited under accession number NCIMB 42382 may be a daughter strain (progeny) or a strain cultured (subcloned) from the original. A tive of a strain of the invention may be modified, for e at the genetic level, without ablating the biological ty. In particular, a derivative strain of the invention is eutically active. A derivative strain will have comparable immune modulatory activity to the al NCIMB 423 82 strain. In particular, a derivative strain will elicit comparable effects on the neurodegenerative disease models and comparable effects on cytokine levels to the effects shown in the Examples, which may be identified by using the culturing and administration protocols described in the Examples. A derivative of the NCIMB 42382 strain will generally be a biotype of the NCIMB 423 82 strain. nces to cells of the Parabacteroides distasonis strain ted under accession number NCIMB 423 82 encompass any cells that have the same safety and therapeutic efficacy characteristics as the strains deposited under ion number NCIMB 42382, and such cells are encompassed by the invention.

In preferred embodiments, the bacterial strains in the compositions of the invention are viable and capable of partially or totally colonising the intestine.

The inventors have found that Parabacteroides distasonis strains reduce the secretion of pro- inflammatory cytokines such as IL-6 and IL-8. IL-8 has been implicated in myelin sheath formation.

Chronic ation induced by IL-6 can ultimately lead to cell death. Therefore, the bacterial strains of the invention are particularly useful in the treatment or prevention of egenerative disorders.

In some embodiments, the ial strains are useful in the treatment of ions characterised by the enhanced activation of IL-6. In some embodiments, the itions of the invention are for use in the treatment or prevention of neurodegenerative diseases characterised by demyelination. Many egenerative es are characterised by ination. Demyelination impedes the propagation of action potentials within neurons, impairing effective communication within the nervous increasystenfij has been shown to contribute positively to myelin sheath formation and repair. MRx0029er se secretion of IL-8. Therefore, the compositions of the invention are particularly [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm ed set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ial in the treatment or tion of neurodegenerative disorders characterised by demyelination.

The inventors have also found that the bacterial strains of invention increase the activation of BDNF.

BDNF is a neurotrophic growth factor that has been shown to enhance neuron entiation and survival. Thus, the compositions of the invention can be used in a method of enhancing nerve cell survival in the treatment or prevention of neurodegenerative diseases.

A further bacteria that may be used in the compositions of the invention is the species Megasphaera massiliensis. The examples demonstrate that Parabacteroides distasom's and haera massiliensis both have neuroprotective ties, but produce different metabolites and may have different mechanisms of action and specific neuroprotective activities. Therefore, these species may be particularly effective when used in combination. In preferred embodiments, the composition comprises a strain of the species Parabacteroides distasonis and a strain of the species Megasphaera massiliensis.

The Parabacteroz'des distasonis bacterium deposited under accession number NCIMB 42382 was tested in the Examples and is also referred to herein as strain 5. MRX0005, MRX005, MRx005 and MRx0005 are used herein interchangeably. A 16S rRNA sequence for the MRx0005 strain that was tested is provided in SEQ ID NO:9. Strain MRx0005 was deposited with the international tary authority NCIMB, Ltd. (Ferguson Building, Aberdeen, AB21 9YA, Scotland) by GT ics Ltd. (Life Sciences Innovation ng, Aberdeen, AB25 2ZS, nd) on 12th March 2015 as "Parabacteroides Sp 755" and was assigned accession number NCIMB 42382. GT Biologics Ltd. Subsequently changed its name to 4D Pharma Research Limited.

The Megasphaera massilz'ensis strain deposited under accession number NCIMB 42787 is the exemplary MRx0029 strain tested in the examples and shown to be effective for treating disease. Strain NCIMB 42787 was deposited with the international depositary authority NCIMB, Ltd. (Ferguson Building, Aberdeen, AB21 9YA, Scotland) by 4D Pharma Research Ltd. (Life Sciences Innovation Building, Aberdeen, AB25 2ZS, nd) on 13th July 2017 as "Megasphaera massiliensis MRx0029" and was assigned accession number NCIMB 42787. Therefore, the invention es a cell, such as an isolated cell, of the Megasphaera massiliensis strain deposited under accession number NCIMB 42787, or a derivative f. The invention also provides a composition comprising a cell of the Megasphaera massiliensis strain deposited under accession number NCIMB 42787, or a tive thereof. The invention also provides a biologically pure culture of the Megasphaera massiliensis strain deposited under accession number NCIMB 42787. The invention also provides a cell of the Megasphaera massiliensis strain deposited under accession number NCIMB 42787, or a derivative f, for use in therapy, in particular for the diseases bed herein.

In odiments, the invention provides a composition comprising the strain deposited at NCIMB under accession number NCIMB 42787, or a derivative or biotype thereof, and the strain deposited at [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm ation] kjm ed set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm ation] kjm ed set by kjm NCIMB under accession number NCIMB 42382, or a tive or biotype thereof, preferably for use in therapy, preferably for use in treating a neurodegenerative disease such as Parkinson’s e.

Therapeutic uses As demonstrated in the examples, the bacterial compositions of the invention are effective for treating neurodegenerative disorders. In particular, treatment with itions of the invention increase neuroproliferation and act as a rotectant against agents that destroy dopaminergic neurons.

Therefore, the compositions of the invention may be useful for treating or preventing neurodegenerative disorders that are the result of neuron death.

Compositions of the invention can decrease the activation of the NFKB promoter, which activates cytokine production, for example IL-lB, IL—loc, IL-l8, TNFor and IL-6. Treating cells with mutant or- synuclein is a model for familial son’s. A point mutation at position 53 from adenine to ine leads to or—synuclein mis-folding. The incorrectly folded or-synuclein subsequently aggregates into insoluble fibrils which form Lewy bodies. Therefore, the compositions of the invention may be useful for treating or preventing egenerative disorders that are the result of neuroinflammation, protein misfolding and/or environmental exposure. Compositions of the invention can be used for treatment of familial Parkinson’s. Activation of the NFKB promoter is mediated through the TLR4 ligand. TL4 is known to mediate cell death in the mouse model MPTP, which simulates Parkinson’s disease.

Compositions of the invention can be used to inhibit the ability of TLR4 signalling to activate the NFKB promoter. Of particular relevance for PD, both TLR2 and TLR4 were found to be upregulated in brains of PD patients [25]. Moreover (x-syn has been described as a ligand for TLR2 [26] and we have demonstrated that or-syn is also a ligand for TLR4 using HEK-TLR4 cells.

Compositions of the invention decrease the secretion of pro-inflammatory cytokines such as IL-6, which can be induced by lipopolysaccharide (LPS). Treatment of cells with LPS tes Parkinson’s caused by environmental factors. Compositions of the invention can be used to decrease IL-6 ion.

Compositions of the invention can be used for treatment of environmental son’s.

Chemokines have been postulated to have important functions in the central nervous system (CNS), in on to their principal role of directional ion of leukocytes. In a murine oligodendrocyte precursor-like cell line the chemokine MCP-l did not increase endrocyte precursor proliferation.

In primary myelinating cultures MCP-l did not enhance myelin segment formation in this system. The inventors have found that MRx0005 es MCP-l levels. In certain embodiments, the compositions of the invention are for use in increasing MCP-l levels in the treatment of disease.

Examples of neurodegenerative diseases to be treated by compositions of the invention include: Parki ’s disease, including progressive supranuclear palsy, progressive supranuclear palsy, Steele- Richa wski syndrome, normal pressure hydrocephalus, vascular or arteriosclerotic [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm parkinsonism and drug-induced parkinsonism; Alzheimer’s disease, including Benson's me; le sclerosis; Huntington’s disease; amyotrophic l sclerosis; Lou Gehrig's disease; motor neurone disease; prion disease; spinocerebellar ataxia; spinal muscular atrophy; ia, including Lewy body, vascular and frontotemporal dementia; primary progressive aphasia; mild cognitive impairment; HIV-related cognitive impairment, and corticobasal ration. A further neurodegenerative diseases to be treated by compositions of the invention is progressive inflammatory neuropathy.

In certain embodiments, the compositions of the invention can be effective for treating neurodegenerative disorders that occur in y patients. The examples show that compositions of the invention can treat Parkinson’s disease which is predominantly sed in patients aged over 65 years old. In preferred embodiments, the itions of the invention are for treating patients 65 years or older. In other certain ments, the patients are n 40 to 65 years old. In other embodiments, the patients are older than 40 years. In certain embodiments, the compositions of the invention are for use in treating a disease associated with old age, for example, a disease diagnosed after 50 years of age.

In certain embodiments, the compositions of the invention are for use in treating a neurodegenerative disorder mediated or terised by the accumulation of protein, in particular mis-folded protein.

In n embodiments, the compositions of the invention are for use in treating a neurodegenerative disorder associated with grey matter neuronal loss. In certain embodiments, the compositions of the invention are for ng a neurodegenerative disorder that is not associated with white matter s.

In certain embodiments, the compositions of the invention are for use in ng a neurodegenerative disorder associated with permanent symptoms.

In certain embodiments, the compositions of the ion are for use in treating a neurodegenerative disorder that is not an auto-immune disorder. In n embodiments, the compositions of the invention are for use in treating a neurodegenerative disorder that is not multiple sclerosis.

In certain embodiments, the compositions of the invention are for use in reducing neuron death, in particular, in the treatment of neurodegenerative ers. In certain embodiments, the compositions of the invention are for use in protecting neurons, in particular in the treatment of neurodegenerative disorders.

The neuroprotective properties of the compositions of the invention, as shown in the examples, mean that the itions may be particularly effective for preventing or delaying onset or progression of neurodegenerative disorders. In certain embodiments, the compositions of the invention are for use in delaying onset or progression of a neurodegenerative disorders. ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Compositions of the invention can increase the secretion of IL-8. IL-8 has been shown to play a role in neuron myelination. In some embodiments, compositions of the invention can be used to increase IL-8 ion.

The therapeutic compositions of the invention can increase the activation of BDNF. BDNF acts on certain neurons of the central nervous system to support the survival of existing neurons and help the growth and development of new neurons and synapses. BDNF is active in the ampus, cortex and basal forebrain, and is important for long-term memory. The compositions of the invention can therefore be used to increase the secretion of BDNF. The itions may therefore be used in the treatment of egenerative diseases associated with the impairment of long-term memory. The compositions of the invention may be used for improving long-term memory, in particular for improving long-term memory that is impaired by a neurodegenerative disease.

In certain embodiments, the compositions ofthe invention increase the mitochondria metabolic activity in neuronal cells.

Modulation oft/w microbiota-gut—bmin axis Communication between the gut and the brain (the microbiota-gut-brain axis) occurs via a bidirectional umoral communication system. Recent evidence shows that the microbiota that resides in the gut can modulate brain development and produce behavioural phenotypes via the microbiota-gut-brain axis. , a number of reviews suggest a role of the iota-gut-brain axis in maintaining central s system functionality and implicate dysfunction of the microbiota-gut-brain axis in the development of central nervous system disorders and conditions [16-27].

The bidirectional communication between the brain and the gut (z'. e. the-gut—brain axis) includes the central nervous system, neuroendocrine and neuroimmune systems, including the hypothalamus- pituitary—adrenal (HPA) axis, sympathetic and parasympathetic arms of the autonomic nervous system (ANS), including the enteric nervous system (ENS) and the vagus nerve, and the gut microbiota.

As demonstrated in the examples, the compositions of the present invention can modulate the microbiota-gut-brain axis and reduce cell death associated with neurodegenerative disorders.

Accordingly, the compositions of the invention may be useful for treating or preventing neurodegenerative disorders, in particular those ers and conditions associated with ction of the iota-gut—brain axis.

In particular embodiments, the compositions of the invention may be useful for treating or preventing a disease or ion selected from the group consisting of: Parkinson’s disease, ing progressive supranuclear palsy, progressive supranuclear palsy, -Richardson-Olszewski me, normal pressure hydrocephalus, ar or arteriosclerotic parkinsonism and drug-induced parkinsonism; Alzheiq’s disease, including Benson's syndrome; multiple sclerosis; Huntington’s disease; amyotrop ic lateral sclerosis; Lou Gehrig's disease; motor neurone disease; prion disease; [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm ionNone set by kjm [Annotation] kjm Unmarked set by kjm spinocerebellar ; spinal muscular atrophy; dementia; including Lewy body; vascular and frontotemporal dementia; y progressive aphasia; mild cognitive impairment; HIV-related cognitive impairment and obasal degeneration.

The compositions of the ion may be particularly useful for ng or preventing chronic disease, treating or preventing disease in patients that have not responded to other therapies (such as treatment with Levodopa, dopamine agonists, MAO-B inhibitors, COMT inhibitors, Glutamate antagonists, and/or anticholinergics), and/or treating or preventing the tissue damage and symptoms associated with dysfunction of the microbiota—gut-brain axis.

In certain embodiments, the compositions of the invention modulate the CNS. In some embodiments, the itions of the invention modulate the autonomic nervous system (ANS). In some embodiments, the compositions of the invention modulate the enteric nervous system (ENS). In some embodiments, the compositions of the invention modulate the hypothalamic, pituitary, adrenal (HPA) axis. In some ments, the compositions of the ion modulate the neuroendocrine pathway.

In some embodiments, the itions of the invention modulate the neuroimmune pathway. In some embodiments, the compositions of the invention modulate the CNS, the ANS, the ENS, the HPA axis and/or the neuroendocrine and neuroimmune pathways. In certain embodiments, the compositions of the invention module the levels of commensal metabolites and/or the gastrointestinal permeability of a t. In certain embodiments, the compositions of the invention may be used to modulate the dopaminergic system.

The signalling of the microbiota—gut-brain axis is modulated by neural s. Accordingly, in some embodiments, the compositions of the invention modulate signalling in neural systems. In certain embodiments, the compositions of the invention modulate the signalling of the central nervous system.

In some embodiments, the compositions of the invention modulate signalling in sensory neurons. In other embodiments, the compositions of the ion modulate ling in motor neurons. In some embodiments, the compositions of the invention modulate the signalling in the ANS. In some embodiments, the ANS is the parasympathetic nervous system. In preferred embodiments, the compositions of the invention te the signalling of the vagus nerve. In other ments, the ANS is the sympathetic nervous system. In other embodiments, the compositions of the invention modulate the signalling in the enteric nervous system. In certain embodiments, the signalling ofANS and ENS neurons responds directly to luminal contents of the gastrointestinal tract. In other embodiments, the ling ofANS and ENS s responds indirectly to neurochemicals produced by luminal bacteria. In other embodiments, the signalling of ANS and ENS neurons responds to neurochemicals produced by luminal bacteria or endocrine cells. In certain preferred embodiments, the neurons of the ENS activate vagal afferents that ce the functions of the CNS.

In some embodiments, the compositions of the ion regulate the activity of enterochromaffin cells. D [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm ed set by kjm Neama’egenerative es Tauopathies are egenerative diseases associated with the pathological aggregation of tau protein in neurofibrillary or gliofibrillary tangles in the human brain. Alzheimer’s disease is an example of a tauopathology. einopathies (also called ou-Synucleinopathies) are neurodegenerative diseases characterised by the abnormal accumulation of aggregates of a-synuclein in s, nerve fibres or glial cells. Parkinson’s disease is an example of a synucleinopathology.

There is clinical and pathological p between these two pathologies. Parkinson's disease patients frequently have dementia and mer's disease patients often manifest parkinsonism [28]. For example, progressive supranuclear palsy (also known as Steele-Richardson—Olszewski syndrome) has a tauopathology, but also leads to prominent parkinsonism [29]. Mutations in LRRKZ known to cause parkinsonism are associated with the accumulation of synuclein, tau, neither, or both proteins [30].

Lewy body disease (LED) is a neurodegenerative disease that is one of the most common causes of dementia in the elderly. LBD exemplifies the existence of a continuum between tau- and synuclein- pathologies. LBD shares clinical and pathological es with Parkinson disease, Parkinson disease dementia and Alzheimer disease [28].

In particular embodiments, the compositions of the invention may be useful for treating or preventing tauopathies and/or synucleinopathies. In particular embodiments, the itions of the invention may be useful for treating or preventing thies. In particular embodiments, the itions of the invention may be useful for treating or preventing synucleinopathies. In certain embodiments, the compositions of the invention may be useful for treating or preventing a disease or condition selected from the group consisting of: Parkinson’s disease, including ssive supranuclear palsy, ssive supranuclear palsy, Steele-Richardson—Olszewski syndrome, normal pressure hydrocephalus, vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism; Alzheimer’s disease, including Benson's syndrome; and dementia; including Lewy body; vascular and frontotemporal dementia.

In preferred embodiments, the compositions of the invention may be useful for ng or preventing Parkinson’s disease, including progressive supranuclear palsy, progressive supranuclear palsy, Steele- Richardson—Olszewski syndrome, normal pressure hydrocephalus, vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism. In preferred ments, the compositions of the ion may be useful for treating or preventing Alzheimer’s disease, ing Benson's syndrome.

In further preferred embodiments, the compositions of the invention may be useful for treating or preventing dementia; including Lewy body; vascular and frontotemporal dementia.

Parkinson ’s disease Parkings disease is a common neurodegenerative disease neuropathologically characterised by 35 degen on of heterogeneous populations of neural cells (dopamine-producing cells). The clinical [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm diagnosis ofParkinson’s disease requires bradykinesia and at least one ofthe following core symptoms: resting tremor; muscle rigidity and postural reflex impairment. Other signs and symptoms that may be present or develop during the progression of the disease are autonomic disturbances (sialorrhoea, seborrhoea, constipation, micturition disturbances, sexual functioning, orthostatic nsion, hyperhydrosis), sleep disturbances and disturbances in the sense of smell or sense of temperature.

Parkinson’s disease is a egenerative diseases that may develop or persist due to dysfunction of the iota-gut-brain axis. Therefore, in preferred embodiments, the compositions of the invention are for use in treating or ting Parkinson’s disease in a subject.

In further red embodiments, the invention provides a composition comprising a bacterial strain of the genus Parabacteroides, for use in a method of ng or preventing Parkinson’s disease.

Compositions sing a bacterial strain of the genus Parabacteroides may improve motor and cognitive functions in models of Parkinson’s disease. Treatment with Parabacteroides strains may modulate ling in the central, autonomic and enteric nervous systems; may modulate the activity of the HPA axis pathway; may te neuroendocrine and/or neuroimmune pathways; and may modulate the levels of commensal metabolites, inflammatory markers and/or gastrointestinal permeability of a subject, all of which are implicated in the neuropathology of Parkinson’s disease. In preferred embodiments, the invention provides a composition comprising a ial strain of the species Parabacteroides distasonis for use in a method of treating or preventing Parkinson’s disease.

Compositions using Parabacteroides distasonis may be particularly effective for treating Parkinson’s disease.

In preferred ments, the compositions of the invention t, reduce or alleviate one or more of the symptoms of Parkinson’s disease in a subject. In preferred embodiments, the compositions of the ion prevent, reduce or alleviate one or more core symptoms of Parkinson’s disease in a subject. In certain embodiments, the compositions of the invention prevent, reduce or ate bradykinesia in a subject. In certain embodiments, the compositions of the invention prevent, reduce or alleviate resting tremor; muscle rigidity and/or postural reflex impairment in a subject. In certain embodiments, the compositions of the ion prevent, reduce or alleviate one or more symptoms associated with Parkinson’s disease progression selected from mic disturbances (sialorrhoea, seborrhoea, constipation, micturition disturbances, sexual functioning, orthostatic hypotension, hyperhydrosis), sleep disturbances and disturbances in the sense of smell or sense of temperature.

In preferred embodiments, the compositions of the invention prevent, reduce or alleviate depressive ms id with Parkinson’s disease. In certain embodiments, the compositions of the invention improve verbal memory and/or executive functions. In certain ments, the compositions of the invention improve attention, g , verbal fluency and/or anxiety.

In othureferred ments, the compositions of the invention prevent, reduce or alleviate cognit ysfunctions comorbid with Parkinson’s disease.

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm In n embodiments, the compositions of the invention prevent, reduce or alleviate Parkinson’s disease progression. In certain embodiments, the compositions of the invention prevent, reduce or alleviate later motor cations. In certain embodiments, the compositions of the invention prevent, reduce or alleviate late motor tions. In certain embodiments, the compositions of the invention prevent, reduce or alleviate neuronal loss. In certain embodiments, the compositions of the invention improve symptoms ofParkinson’s disease dementia (PDD). In n embodiments, the itions ofthe invention prevent, reduce or alleviate impairment of executive on, attention and/or working memory. In certain embodiments, the compositions of the invention improve dopaminergic neurotransmission. In certain ments, the compositions of the invention prevent, reduce or alleviate impaired dopaminergic neurotransmission.

In some embodiments, the compositions of the invention improve the symptoms of Parkinson’s e according to a symptomatic or diagnostic scale. In certain embodiments, the tests for assessing symptomatic improvement ofmotor on in Parkinson’s disease is the Unified Parkinson’s Disease Rating Scale. In particular, UPDRS II considers the activity of daily life and UPDRS III considers motor-examination.

In some ments, the compositions of the invention improve the ms associated the PDD according to a matic or diagnostic test and/or scale. In certain embodiments, the test or scale is selected from the Hopkins Verbal Learning Test — Revised R); the Delis-Kaplan Executive on System (D-KEFS) Color-Word Interference Test; the Hamilton Depression Rating Scale (HAM-D l7; depression); the Hamilton Anxiety Rating Scale (HAM—A; anxiety) and the Unified Parkinson’s Disease Rating Scale (UPDRS; PD symptom severity).

In some embodiments, the compositions of the invention improve the Clinical Global Impression — Global Improvement (CGI—I) scale for assessing atric and neurological disorders. In some ments, the compositions of the invention display a positive effect on global social and occupational impairment of the subject with Parkinson’s disease.

Alzheimer ’s disease and dementia In DSM-5, the term dementia was replaced with the terms major neurocognitive er and mild ognitive disorder. Neurocognitive disorder is a heterogeneous class of psychiatric diseases. The most common neurocognitive disorder is Alzheimer’s disease, followed by vascular dementias or mixed forms of the two. Other forms of neurodegenerative disorders (e.g. Lewy body disease, frontotemporal dementia, Parkinson’s dementia, Creutzfeldt-Jakob disease, Huntington’s disease, and Wernicke-Korsakoff syndrome) are accompanied by dementia.

Alzheimer’s disease and dementia are also terised by neuronal loss, so the neuroprotective and neuroproliferative effects shown in the examples for the compositions of the invention te that they true useful for treating or preventing these conditions.

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm ed set by kjm The matic criteria for dementia under DSM-5 are evidence of significant cognitive decline from a previous level ofperformance in one or more cognitive domains selected from: learning and memory; language; executive function; complex attention; perceptual-motor and social cognition. The cognitive s must interfere with independence in everyday activities. In on, the cognitive deficits do not occur exclusively in the context of a delirium and are not better explained by another mental er (for example MDD or schizophrenia).

In on to the primary symptom, subjects with neurodegenerative ers y behavioural and psychiatric symptoms ing agitation, aggression, sion, anxiety, apathy, psychosis and sleep-wake cycle disturbances.

Neurodegenerative disorders may develop or persist due to dysfunction of the microbiota—gut-brain axis. Therefore, in preferred embodiments, the compositions of the invention are for use in treating or preventing neurodegenerative disorders in a subject. In preferred embodiments, the neurodegenerative disorder is mer’s disease. In other embodiments, the neurodegenerative disorder is selected from ar dementias; mixed form Alzheimer’s disease and vascular dementia; Lewy body disease; frontotemporal dementia; Parkinson’s dementia; Creutzfeldt-Jakob disease; Huntington’s disease; and Wernicke-Korsakoff me.

In preferred embodiments, the compositions of the ion prevent, reduce or alleviate one or more of the symptoms ofneurodegenerative disorders in a subject. In certain embodiments, the compositions of the invention prevent, reduce or alleviate the occurrence of cognitive decline in a subject. In certain embodiments, the compositions of the invention improve the level of performance of a subject with neurodegenerative disorders in one or more cognitive domains selected from: learning and memory; language; executive function; complex attention; perceptual-motor and social cognition. In some embodiments, the compositions of the invention prevent, reduce or alleviate the occurrence of one or more behavioural and psychiatric ms associated with neurodegenerative disorders selected from agitation, aggression, depression, anxiety, apathy, psychosis and sleep-wake cycle disturbances.

In certain embodiments, the compositions of the invention prevent, reduce or alleviate matic disease by intervention in suspected pathogenic mechanisms at a preclinical stage. In certain embodiments, the compositions of the invention improve disease ation, with slowing or arrest of m progression. In some embodiments, the slowing or arrest of symptom progression correlates with evidence in ng the underlying neuropathological process. In preferred embodiments, the compositions of the invention improve symptoms of neurodegenerative disorders comprising enhanced ive and functional improvement. In preferred embodiments, the compositions of the invention e the oural and psychiatric symptoms of dementia (BPSD).

In preferred embodiments, the compositions of the invention improve the ability of a subject with neurodfinerative disorder to undertake everyday activities.

[Annotation] kjm None set by kjm [Annotation] kjm ionNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm In preferred embodiments, the compositions of the invention improve both ion and functioning in a subject with mer’s disease. In some embodiments, the composition of the invention improves the cognitive endpoint in a subject with Alzheimer’s disease. In some embodiments, the itions of the ion improve the functional endpoint in a t with mer’s disease.

In preferred embodiments, the compositions of the invention improve the cognitive and functional endpoint in a subject with mer’s disease. In yet further preferred ments, the compositions of the invention improve the overall clinical response (the global endpoint) in a subject with Alzheimer’s e.

In some embodiments, the compositions of the invention improve the symptoms of neurodegenerative ers according to a symptomatic or diagnostic test. In certain ments, the tests for assessing symptomatic improvement of Alzheimer’s disease (and other neurodegenerative disorders) are selected from objective cognitive, activities of daily living, global ment of change, health related quality of life tests and tests assessing behavioural and psychiatric ms of neurodegenerative disorders.

In certain embodiments, the objective cognitive tests for assessment of matic improvement use the Alzheimer’s disease Assessment Scale cognitive subscale (ADAS-cog) and the classic ADAS scale. In certain embodiments, symptomatic improvement of cognition is assessed using the Neurophysiological Test Battery for Use in Alzheimer’s Disease (NTB).

In some embodiments, the global assessment of change test uses the Clinical Global Impression — Global Improvement (CGI—I) scale for assessing psychiatric and neurological disorders. In some embodiments, the global scale is the Clinician's Interview Based Impression of Change plus (CIBIC-plus). In some embodiments, the global scale is the Alzheimer’s e Cooperative Study Unit Clinician’s Global Impression of Change (ADCS-CGIC).

In certain embodiments, the health related quality of life measures are the Alzheimer’s Disease-Related QOL (ADRQL) and the QOL-Alzheimer’s Disease (QOL-AD).

In certain embodiments, the tests assessing behavioural and psychiatric symptoms of neurodegenerative disorders are selected from the Behavioural pathology in Alzheimer’s Disease Rating Scale (BEHAVE-AD); the Behavioural Rating Scale for Dementia (BRSD); the Neuropsychiatric Inventory (NPI); and the Cohen-Mansfield Agitation Inventory .

In some embodiments, the compositions of the invention are particularly effective at preventing, reducing or alleviating neurodegenerative disorders when used in combination with r y for treating neurodegenerative disorders. In certain embodiments, such therapies include cholinesterase inhibitors including donepezil (Aricept®), galantamine (Razadyne®) and rivastifiue (Exelon ®), and memantine.

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Multiple Sclerosis Multiple sclerosis (MS) is a demyelinating disease in which the myelin sheath surrounding neurons in the brain and spinal cord are damaged. The exact underlying causes of MS are unknown, but are thought to vary between individuals. Certain forms of MS are hereditary. Environmental factors are also thought to bute to MS. In some individuals, a combination ofboth c and environmental factors may trigger the onset of MS.

There are a wide variety of ms associated with MS. ts may exhibit almost any neurological symptom associated with the impairment of autonomic, Visual, motor or sensory l.

The exact symptoms will vary depending on the site of neuronal damage/demyelination.

IL-8 has been implicated in the formation of myelin sheaths. The compositions of the invention may ore be for use in the remyelination of neurons in ts with MS. The compositions of the ion may also be used to protect neurons from ination. In other words, the compositions of the invention may be for use in a method of treating or preventing multiple sis by restoring or preventing loss of neuron myelin sheaths.

In some embodiments, the compositions of the ion prevent, reduce or alleviate one or more symptoms ofMS in a subject. In some embodiments, the compositions of the invention t, reduce or alleviate fatigue in a subject. In certain embodiments, the compositions of the invention prevent, reduce or alleviate resting tremor, muscle weakness, muscle spasms, muscle stiffness, paraesthesia and/or ataxia in a subject. In n embodiments, the compositions of the invention prevent, reduce or alleviate one or more symptoms associated with MS progression selected from the list consisting of autonomic disturbances: constipation, micturition disturbances, sexual functioning, dysphagia, dysarthria, syncope, vertigo and/or dizziness; sleep disturbances; and disturbances in the sense of smell or sense of temperature. In some embodiments, the compositions of the invention prevent, reduce or alleviate one or more ocular symptoms associated with MS. In some embodiments, the ocular symptom is ed from the list consisting of loss of , eye pain, colour blindness, double Vision and/or involuntary eye movements in a subject.

In some embodiments, the itions of the invention prevent, reduce or alleviate dizziness, vertigo, neuropathic pain, musculoskeletal pain, cognitive dysfunction, bowel inence, dysphagia, dysarthria, or any combination thereof.

In some embodiments, the compositions of the invention prevent, reduce or alleviate depressive symptoms or anxiety comorbid with MS.

In some embodiments, the improvement of symptoms are determined using the 2017 McDonald ia for diagnosing MS.

In certDembodiments, treatment with the compositions of the invention results in a reduction in MS incidence or MS severity. In certain embodiments, the compositions of the invention are for use in [Annotation] kjm None set by kjm ation] kjm ionNone set by kjm ation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm ation] kjm Unmarked set by kjm reducing relapse incidence or relapse severity. In certain embodiments, ent with the compositions of the invention prevents a decline in motor function or results in improved motor function associated with MS. In certain embodiments, the compositions of the invention are for use in ting a decline in motor function or for use in ing motor function in the treatment of MS.

In certain embodiments, treatment with the compositions of the invention prevents the development of paralysis in MS. In n embodiments, the compositions of the invention are for use in preventing paralysis in the treatment of MS.

In certain embodiments the compositions of the invention are for use in preventing multiple sclerosis in a patient that has been identified as at risk of multiple sis, or that has been diagnosed with early—stage multiple sclerosis or "relapsing-remitting" multiple sclerosis. The compositions of the invention may be useful for preventing the development of MS. The compositions of the ion may be useful for preventing the progression of MS. In certain embodiments, the compositions of the invention are for use in a patient identified as having a c predisposition to MS, such as major histocompatibility complex (MHC) class II phenotype, human leukocyte antigen (HLA)-DR2 or HLA- DR4.

The compositions of the invention may be useful for ng or alleviating multiple sclerosis. The compositions of the ion may be particularly useful for reducing symptoms associated with le sis. Treatment or prevention of multiple sclerosis may refer to, for example, an ation of the severity of symptoms or a reduction in the frequency of exacerbations or the range of triggers that are a problem for the patient. In certain embodiments, the compositions of the invention slow or stop progression of the disease.

In certain embodiments, the compositions of the invention are for use in treating ing-remitting MS. In alternative embodiments, the compositions of the invention are for use in treating progressive MS, such as secondary progressive MS (SPMS), which develops over time following diagnosis of RRMS, y progressive MS (PPMS) which exhibits gradual continuous neurologic deterioration and progressive relapsing MS (PRMS), which is similar to PPMS but with overlapping relapses.

In certain embodiments, the compositions of the invention are for use in treating one or more of symptoms ofMS selected from the group consisting of: fatigue, vision problems, numbness, tingling, muscle spasms, muscle stiffness, muscle weakness, mobility problems, pain, ms with thinking, learning and planning, depression and anxiety, sexual problems, bladder problems, bowel problems, speech and swallowing difficulties.

Neurochemical factors, neuropeptides and neurotransmitters and the microbiota-gut—brain axis As outlined above, the microbiota—gut-brain axis is modulated by a number of different physiological systems. The microbiota—gut—brain axis is modulated by a number of signalling molecules. Alterations in the gels of these signalling molecules results in neurodegenerative diseases. The experiments performed by the inventors indicate that administration of Parabacteroides s, and in particular [Annotation] kjm None set by kjm [Annotation] kjm ionNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Parabacteroides distasonis, can modulate levels of indole and kynurenine. ulation of these metabolites can lead to egenerative diseases, such as Parkinson’s disease.

In certain embodiments, the compositions of the invention modulate the levels of brain monoamines and metabolites f. In red embodiments the metabolite is kynurenine. In n embodiments, the compositions of the invention te kynurenine, which is the main route of phan metabolism, which serves as a route to namide adenine dinucleotide (NAD+) production. Kynurenine can be metabolized to neuroactive compounds such as nic acid (KYNA) and 3-hydroxy-l-kynurenine (3-OH-l-KYN), and in further steps to quinolinic acid (QUIN).

Dysregulation of the kynurenine pathway can lead to activation of the immune system and the accumulation of potentially neurotoxic compounds. Alterations in the kynurenine metabolism may be involved in the development of Parkinson’s diseases. Kynurenine levels have been demonstrated to be decreased in the frontal cortex, putamen and substantia nigra pars compacta of patients with PD [31].

Therefore, in certain embodiments the itions of the invention are for use in increasing the levels of kynurenine in the treatment of Parkinson’s disease.

In certain embodiments of the invention the compositions of the invention can increase the levels of kynurenin. Increased levels of kynurenine have been shown to attenuated MPP+-induced neuronal cell death in vitro in a human dopaminergic neuroblastoma cell line [32]. In certain embodiments kynurenine and kynurenic acid, can activate GI aryl arbon receptor (Ahr) and GPR35 receptors.

Activation ofAhr receptor induces IL-22 production, which can inhibit local inflammation. Activation of GPR35 inducing the production of inositol triphosphate and Ca2+ mobilization.

In certain embodiments, the itions of the invention modulate the levels of . In red embodiments the lite is kynurenine. In certain embodiments, the compositions of the invention modulate nine routes, which is the main route of tryptophan metabolism.

The signalling of the microbiota—gut-brain axis is modulated by levels of neurochemical factors, neuropeptides and neurotransmitters. Accordingly, in certain embodiments, the compositions of the invention modulates levels of neurochemical factors, neuropeptides and neurotransmitters. ingly, in certain preferred embodiments, the compositions of the invention directly alter CNS biochemistry.

The signalling of the microbiota—gut-brain axis is modulated by levels of y-aminobutyric acid (GABA).

Accordingly, in preferred embodiments, the itions of the invention modulate the levels of GABA. GABA is an inhibitory neurotransmitter that reduces neuronal excitability. In certain embodiments, the compositions of the invention increase the levels of GABA. In n embodiments, the compositions of the invention decrease the levels of GABA. In certain embodiments, the compositions of the invention alter GABAergic neurotransmission. In certain embodiments, the compchs of the invention modulate the level of GABA transcription in different regions of the central nervous system. In certain embodiments, the commensal derived GABA crosses the blood- [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm ed set by kjm brain r and affects neurotransmission directly. In certain embodiments, the compositions of the invention lead to a reduction of GABA in the hippocampus, amygdala and/or locus coeruleus. In certain embodiments, the compositions of the invention lead to an increase of GABA in cortical regions.

Immune res onse The signalling of the microbiota-gut—brain axis is modulated by alterations in the immune response and atory factors and markers. Accordingly, in certain embodiments, the compositions of the invention may modulate the immune response. In certain embodiments, the compositions of the invention modulate the systemic levels of circulating mmune signalling molecules. In certain preferred embodiments, the itions of the invention modulate pro-inflammatory cytokine production and inflammation. In certain embodiments, the compositions of the ion modulate the atory state. In certain embodiments, the compositions of the ion decrease IL-6 production and secretion. In certain embodiments, the compositions of the invention decrease the activation of the NFKB promoter. In n ments, the compositions of the invention are able to modulate the activation of IL-6 production by the potent pro-inflammatory endotoxin lipopolysaccharide (LPS). In certain embodiments, the compositions of the invention are able to modulate the activation of the NFKB promoter by LPS and or-synuclein mutant proteins such as A53T.

Increased ating levels of cytokines are closely associated with various neurodegenerative disorders, including Parkinson’s, dementia and Alzheimer’s. In certain embodiments, the compositions of the invention are for use in reducing IL-6 levels and/or NFKB levels in the treatment of a neurodegenerative disorder.

In some ments, the compositions of the invention increase the secretion of IL-8. IL-8 has been shown to induce myelin sheath formation and restore or preserve effective neuronal communication.

Thus, in some embodiments, the compositions of the invention are for use in inducing myelin sheath formation in the treatment of neurodegenerative diseases. In some embodiments, the compositions of the invention are for use in restoring al communication. In some embodiments, the compositions of the invention are for use in preserving neuronal communication.

The signalling of the microbiota-gut—brain axis is ted by levels of commensal metabolites.

Accordingly, in certain ments, the compositions of the invention modulate the systemic levels of microbiota metabolites. In certain preferred embodiments, the itions of the invention te the level of short chain fatty acids ). In certain embodiments the level of SCFAs is increased or decreased. In some embodiments, the SCFA is butyric acid (BA) (or butyrate). In some embodiments, the SCFA is propionic acid (PPA). In some embodiments, the SCFA is acetic acid. In certain embodiments, the compositions of the invention modulate the y of SCFAs to cross the blood—fin barrier. ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Histone acetylation and deacetylation are important epigenetic regulators of gene expression. An imbalance in histone acetylation and deacetylation can result in apoptosis. Dysregulation of such histone acetyltransferases has been implicated in the pathogenesis associated with age-associated neurodegenerative es, such as son’s e, gton's disease, Alzheimer's disease, amyotrophic lateral sclerosis and cognitive decline [33]. Accordingly, in n embodiments, the compositions of the invention can modulate e deacetylatase ty. In certain embodiments, the compositions of the invention can reduce histone deacetylatase activity. In certain ments, the compositions of the ion can reduce histone acetylatase activity.

Patients with neurodegenerative diseases, including son’s disease, Huntington's disease, Alzheimer's disease and amyotrophic lateral sclerosis, exhibit high levels of lipid dation. Lipid are vulnerable to ion by reactive oxygen species, and the brain is rich in polyunsaturated fatty acids. Accordingly, in certain ments, the compositions of the ion can modulate lipid peroxidation. In certain embodiments, the compositions of the invention can reduce lipid peroxidation.

Reducing the oxidative damage caused by reactive oxygen species can be used to target early the stages neurodegenerative diseases. ingly, in certain embodiments, the compositions of the invention are for use in treating early stage neurodegeneration. Also accordingly, in certain embodiments, the compositions of the invention are for use in preventing the development of a neurodegenerative disorder. In such embodiments, the compositions of the invention may be for use in a t that has been identified as at risk of developing a neurodegenerative disorder.

The signalling of the microbiota—gut-brain axis is modulated by levels of gastrointestinal permeability.

Accordingly, in some embodiments, the compositions of the invention alter the integrity of the gastrointestinal tract epithelium. In certain embodiments, the compositions of the invention modulate the permeability of the gastrointestinal tract. In certain embodiments, the compositions of the invention modulate the barrier function and integrity of the gastrointestinal tract. In certain embodiments, the compositions of the invention modulate gastrointestinal tract motility. In certain embodiments, the compositions of the invention modulate the translocation of commensal metabolites and inflammatory signalling molecules into the bloodstream from the gastrointestinal tract lumen.

The signalling of the microbiota—gut-brain axis is modulated by microbiome composition in the gastrointestinal tract. Accordingly, in n embodiments, the compositions of the invention modulates the microbiome composition of the gastrointestinal tract. In certain embodiments, the compositions of the invention prevents microbiome sis and associated ses in toxic metabolites (e.g. LPS). In certain ments, the compositions of the invention modulate the levels of Clostridium in the intestinal tract. In preferred embodiments, the compositions of the invention reduce the level of Clostridium in the gastrointestinal tract. In certain embodiments, the compositions of the invention reduce the levels of Campylobacter jejuni. In certain embodiments, the compans of the invention modulate the proliferation of harmful anaerobic bacteria and the production of neurotoxins produced by these bacteria. In certain embodiments, the compositions of the [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm invention modulate the iome levels of Lactobacillus and/or Bifidobacterium. In certain embodiments, the compositions of the ion modulate the microbiome levels of Sutterella, ella, Ruminococcus genera and/or the Alcaligenaceae family. In certain embodiments, the compositions of the ion increase the level of Lactobacillus plantarum and/or Saccharomyces boulardii.

Brain injury The examples demonstrate that the compositions of the invention are neuroprotective and have HDAC inhibitory ty. HDACZ is a crucial target for functional recovery from stroke [34] and HDAC inhibition can prevent white matter injury [35], so the compositions of the invention may be useful in the treatment of brain injury.

In certain embodiments, the compositions of the invention are for use in treating brain injury. In some embodiments, the brain injury is a traumatic brain injury. In some embodiments, the brain injury is an acquired brain injury. In some embodiments, the compositions of the invention are for use in treating brain injury resulting from trauma. In some embodiments, the compositions of the invention are for use in treating brain injury resulting from a tumour. In some embodiments, the compositions of the invention are for use in treating brain injury ing from a stroke. In some embodiments, the compositions of the invention are for use in treating brain injury resulting from a brain haemorrhage.

In some embodiments, the compositions of the invention are for use in treating brain injury resulting from encephalitis. In some embodiments, the compositions of the invention are for use in treating brain injury resulting from cerebral a. In some embodiments, the compositions of the invention are for use in treating brain injury resulting from al anoxia.

In preferred ments, the compositions of the invention are for use in treating stroke. The effects shown in the examples are ularly relevant to the treatment of stroke. Stroke occurs when blood flow to at least a part of the brain is interrupted. Without an adequate supply of blood to provide oxygen and nutrients to the brain tissue and to remove waste products from the brain tissue, brain cells rapidly begin to die. The symptoms of stroke are dependent on the region of the brain which is ed by the inadequate blood flow. Symptoms include paralysis, ss or weakness of the muscles, loss of balance, dizziness, sudden severe headaches, speech impairment, loss of memory, loss of reasoning ability, sudden confusion, vision impairment, coma or even death. A stroke is also referred to as a brain attack or a ovascular accident (CVA). The symptoms of stroke may be brief if te blood flow is restored within a short period of time. However, if inadequate blood flow continues for a significant period of time, the symptoms can be permanent.

In some embodiments, the stroke is cerebral ischemia. Cerebral ischemia results when there is insufficient blood flow to the tissues of the brain to meet metabolic . In some embodiments, the ce 1 ischemia is focal cerebral ischemia, i.e. confined to a specific region of the brain. In some ments the cerebral ischemia is global cerebral ischemia, i.e. encompassing a wide area of the [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm ation] kjm Unmarked set by kjm brain tissue. Focal al ischemia commonly occurs when a cerebral vessel has become blocked, either partially or completely, reducing the flow of blood to a c region of the brain. In some embodiments the focal cerebral ischemia is ischemic stroke. In some embodiments, the ischemic stroke is thrombotic, i.e. caused by a thrombus or blood clot, which develops in a cerebral vessel and restricts or blocks blood flow. In some embodiments the ischemic stroke is a otic stroke. In some embodiments, the ischemic stroke is embolic, i.e. caused by an embolus, or an unattached mass that travels through the bloodstream and restricts or blocks blood flow at a site distant from its point of origin. In some embodiments the ischemic stroke is an embolic stroke. Global cerebral ischemia commonly occurs when blood flow to the brain as a whole is blocked or reduced. In some embodiments the global cerebral ischemia is caused by hypoperfusion, i.e. due to shock. In some embodiments the global cerebral ischemia is a result of a cardiac arrest.

In some embodiments the subject diagnosed with brain injury has suffered cerebral ischemia. In some embodiments, the subject diagnosed with brain injury has suffered focal cerebral ischemia. In some embodiments, the subject diagnosed with brain injury has ed an ischemic stroke. In some embodiments, the subject diagnosed with brain injury has suffered a otic stroke. In some embodiments, the subject diagnosed with brain injury has suffered an embolic stroke. In some embodiments, the subject diagnosed with brain injury has suffered global cerebral ischemia. In some embodiments, the subject diagnosed with brain injury has suffered hypoperfusion. In some embodiments, the subject diagnosed with brain injury has suffered a cardiac arrest.

In some embodiments, the compositions of the invention are for use in treating cerebral ischemia. In some embodiments, the compositions of the invention are for use in treating focal cerebral ischemia.

In some embodiments, the compositions of the invention are for use treating ischemic stroke. In some ments, the compositions of the invention are for use in treating thrombotic . In some embodiments, the compositions of the invention are for use in treating embolic stroke. In some embodiments, the compositions of the invention are for use in treating global cerebral ischemia. In some embodiments, the itions of the ion are for use in treating rfusion.

In some embodiments, the stroke is hemorrhagic stroke. Hemorrhagic stroke is caused by ng into or around the brain ing in swelling, pressure and damage to the cells and s of the brain.

Hemorrhagic stroke is commonly a result of a weakened blood vessel that ruptures and bleeds into the surrounding brain. In some embodiments, the hemorrhagic stroke is an intracerebral hemorrhage, i.e. caused by bleeding within the brain tissue itself. In some ments the intracerebral hemorrhage is caused by an intraparenchymal hemorrhage. In some embodiments the intracerebral hemorrhage is caused by an intraventricular hemorrhage. In some embodiments the hemorrhagic stroke is a chnoid hage i.e. bleeding that occurs outside of the brain tissue but still within the skull.

In some embodiments, the hagic stroke is a result of cerebral amyloid angiopathy. In some embocnnts, the hemorrhagic stroke is a result of a brain aneurysm. In some embodiments, the hemorrhagic stroke is a result of cerebral ovenous malformation (AVM).

[Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm In some embodiments the subject diagnosed with brain injury has suffered hemorrhagic stroke. In some embodiments, the subject diagnosed with brain injury has suffered an intracerebral hemorrhage.

In some ments, the subject diagnosed with brain injury has suffered an intraparenchymal hemorrhage. In some embodiments, the subject diagnosed with brain injury has suffered an intraventricular hemorrhage. In some embodiments, the subject diagnosed with brain injury has suffered a subarachnoid hemorrhage. In some embodiments, the t diagnosed with brain injury has suffered cerebral amyloid athy. In some embodiments, the subject diagnosed with brain injury has suffered a brain sm. In some embodiments, the subject sed with brain injury has suffered cerebral AVM.

In some embodiments, the itions of the invention are for use in treating hemorrhagic stroke. In some embodiments, the compositions of the invention are for use in treating an intracerebral hemorrhage. In some embodiments, the compositions of the invention are for use in treating an intraparenchymal hemorrhage. In some embodiments, the compositions of the invention are for use in treating an intraventricular hemorrhage. In some embodiments, the itions of the ion are for use in treating a subarachnoid hemorrhage. In some embodiments, the compositions of the invention are for use in treating a cerebral amyloid angiopathy. In some embodiments, the compositions of the invention are for use in treating a brain aneurysm. In some embodiments, the compositions of the invention are for use in treating cerebral AVM.

Restoration of adequate blood flow to the brain after a period of interruption, though effective in alleviating the symptoms associated with stroke, can paradoxically result in further damage to the brain . During the period of interruption, the affected tissue suffers from a lack of oxygen and nutrients, and the sudden restoration of blood flow can result in ation and oxidative damage through the induction of oxidative stress. This is known as reperfusion injury, and is well documented not only following stroke, but also following a heart attack or other tissue damage when blood supply returns to the tissue after a period of ischemia or lack of oxygen. In some embodiments the subject diagnosed with brain injury has suffered from reperfusion injury as a result of stroke. In some embodiments, the compositions of the invention are for use in treating reperfusion injury as a result of stroke.

A transient ischemic attack (TIA), often referred to as a mini-stroke, is a ised g sign for a more serious stroke. Subjects who have ed one or more TIAs are therefore at greater risk of stroke. In some embodiments the subject diagnosed with brain injury has suffered a TIA. In some embodiments, the compositions of the invention are for use in treating a TIA. In some embodiments, the compositions of the invention are for use in treating brain injury in a t who has suffered a TIA.

High blood pressure, high blood terol, a familial history of stroke, heart disease, diabetes, brain aneury arteriovenous mations, sickle cell , disease, vasculitis, bleeding ers, use of nonste al anti-inflammatory drugs (NSAIDs), smoking tobacco, drinking large amounts of alcohol, [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm illegal drug use, obesity, lack of physical activity and an unhealthy diet are all considered to be risk s for stroke. In particular, lowering blood pressure has been conclusively shown to prevent both ischemic and hemorrhagic strokes [36, 37]. In some embodiments, the compositions of the invention are for use in treating brain injury in a t who has at least one risk factor for stroke. In some embodiments the subject has two risk factors for . In some embodiments the subject has three risk factors for stroke. In some embodiments the subject has four risk factors for stroke. In some embodiments the subject has more than four risk factors for stroke. In some embodiments the subject has high blood pressure. In some embodiments the subject has high blood cholesterol. In some embodiments the subject has a familial history of stroke. In some embodiments the subject has heart disease. In some embodiments the subject has diabetes. In some embodiments the subject has a brain aneurysm. In some embodiments the subject has arteriovenous malformations. In some embodiments the t has vasculitis. In some embodiments the subject has sickle cell e. In some embodiments the subject has a bleeding disorder. In some embodiments the subject has a history of use of nonsteroidal anti-inflammatory drugs (NSAIDs). In some embodiments the subject smokes tobacco. In some embodiments the subject drinks large amounts of alcohol. In some embodiments the subject uses illegal drugs. In some embodiments the t is obese. In some embodiments the subject is overweight. In some embodiments the subject has a lack of physical activity. In some embodiments the subject has an unhealthy diet.

The examples indicate that the compositions of the invention may be useful for treating brain injury and aiding recovery when stered before the injury event occurs. Therefore, the compositions of the invention may be particularly useful for treating brain injury when administered to subjects at risk of brain injury, such as stroke.

In n embodiments, the compositions of the invention are for use in reducing the damage caused by a potential brain injury, preferably a stroke. The itions may reduce the damage caused when they are stered before the potential brain injury occurs, in particular when administered to a patient identified as at risk of a brain injury.

The es te that the compositions of the invention may be useful for treating brain injury and aiding recovery when stered after the injury event occurs. Therefore, the compositions of the invention may be particularly useful for treating brain injury when administered to subjects following a brain injury, such as stroke.

In some embodiments, the compositions of the invention treat brain injury by reducing c damage. In some embodiments, the compositions of the invention treat brain injury by ing motor function. In some embodiments, the compositions of the invention treat brain injury by improving muscle th. In some embodiments, the compositions of the invention treat brain injury by imfiing memory. In some embodiments, the itions of the invention treat brain injury by [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm improving social recognition. In some embodiments, the compositions of the invention treat brain injury by improving neurological function.

Treatment of brain injury may refer to, for example, an alleviation of the severity of symptoms.

Treatment of brain injury may also refer to reducing the ogical impairments following stroke. itions of the invention for use in treating stroke may be provided to the subject in advance of the onset of stroke, for example in a patient identified as being at risk of stroke. Compositions of the invention for use in treating stroke may be provided after a stroke has occurred, for example, during recovery. Compositions of the invention for use in treating stroke may be provided during the acute phase of recovery (i.e. up to one week after stroke). Compositions of the invention for use in treating stroke may be provided during the subacute phase of recovery (i.e. from one week up to three months after stroke). Compositions of the invention for use in treating stroke may be provided during the chronic phase of recovery (from three months after stroke).

In certain embodiments, the itions of the invention are for use in combination with a ary active agent. In certain embodiments, the itions of the invention are for use in combination with aspirin or tissue plasminogen activator (tPA). Other secondary agents include other antiplatelets (such as clopidogrel), agulants (such as heparins, warfarin, apixaban, dabigatran, an or xaban), pertensives (such as diuretics, ACE inhibitors, calcium channel rs, beta- blockers or alpha-blockers) or statins. The compositions of the invention may improve the patient’s response to the secondary active agent.

In n ments, the compositions of the invention reduce the effect of ischemia on tissues. In certain embodiments, the compositions of the invention reduce the amount of damage to tissues caused by ischemia. In n embodiments, the tissues damaged by ischemia are the cerebral tissues. In certain embodiments, the compositions of the invention reduce necrosis or the number of necrotic cells.

In certain embodiments, the compositions of the invention reduce apoptosis or the number of tic cells. In certain embodiments, the compositions of the invention reduce the number of necrotic and apoptotic cells. In certain embodiments, the compositions of the invention prevent cell death by necrosis and/or apoptosis. In certain embodiments, the compositions of the invention prevent cell death by necrosis and/or apoptosis caused by ischemia. In n ments, the compositions of the invention improve the ry of the tissue damaged by ischemia. In certain embodiments, the compositions of the invention improve the speed of nce of necrotic cells and/or tic cells.

In certain embodiments, the compositions of the invention improve the efficacy of the clearance of necrotic cells and/or apoptotic cells. In certain embodiments, the compositions of the invention improve the replacement and/or regeneration of cells within tissues. In n embodiments, the compositions of the invention improve the replacement and/or regeneration of cells within tissues damaged by ischemia. In certain embodiments, the compositions of the invention e the overall histolrnyf the tissue (for example upon a biopsy).

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Modes ofadministration Preferably, the compositions ofthe invention are to be administered to the gastrointestinal tract in order to enable delivery to and/ or partial or total colonisation of the intestine with the bacterial strain of the invention. Generally, the compositions of the invention are administered orally, but they may be administered rectally, intranasally, or Via buccal or sublingual routes.

In certain embodiments, the compositions of the invention may be administered as a foam, as a spray or a gel.

In certain embodiments, the compositions of the invention may be administered as a suppository, such as a rectal suppository, for example in the form of a theobroma oil (cocoa butter), synthetic hard fat (e.g. ire, witepsol), glycero-gelatin, polyethylene glycol, or soap glycerin composition.

In n embodiments, the composition of the invention is administered to the gastrointestinal tract Via a tube, such as a nasogastric tube, orogastric tube, gastric tube, jejunostomy tube (J tube), aneous endoscopic gastrostomy (PEG), or a port, such as a chest wall port that provides access to the stomach, jejunum and other suitable access ports.

The compositions of the invention may be administered once, or they may be administered sequentially as part of a treatment regimen. In certain embodiments, the compositions of the invention are to be administered daily.

In certain embodiments of the invention, treatment according to the invention is accompanied by assessment of the patient’s gut iota. Treatment may be repeated if delivery of and / or partial or total colonisation with the strain of the invention is not achieved such that efficacy is not observed, or treatment may be ceased if delivery and/ or partial or total colonisation is successful and efficacy is observed.

In certain embodiments, the composition of the invention may be administered to a nt animal, for example a mammal such as a human in order to prevent an inflammatory or mune disease developing in her child in utero and / or after it is born.

The compositions of the invention may be administered to a patient that has been diagnosed with a egenerative disease, or that has been fied as being at risk of a egenerative e.

The compositions may also be administered as a lactic measure to prevent the development of neurodegenerative disease in a healthy patient.

The compositions of the invention may be administered to a patient that has been fied as having an abnormal gut microbiota. For example, the patient may have reduced or absent colonisation by Parabacteroides, and in particular Parabacteroides distasonis.

The ions of the invention may be administered as a food product, such as a nutritional supplement.

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Generally, the compositions of the invention are for the treatment of humans, although they may be used to treat animals including monogastric s such as poultry, pigs, cats, dogs, horses or rabbits. The compositions of the invention may be useful for enhancing the growth and performance of animals. If administered to animals, oral gavage may be used.

Compositions Generally, the composition of the invention comprises bacteria. In preferred embodiments of the invention, the ition is formulated in freeze-dried form. For example, the composition of the ion may comprise granules or gelatin capsules, for example hard gelatin capsules, comprising a bacterial strain of the invention.

Preferably, the composition of the invention comprises lyophilised bacteria. Lyophilisation of bacteria is a stablished procedure and relevant guidance is available in, for example, references [3 8-40].

Alternatively, the composition of the invention may comprise a live, active bacterial culture.

In some ments, the bacterial strain in the composition of the invention has not been inactivated, for e, has not been heat-inactivated. In some embodiments, the bacterial strain in the composition of the invention has not been killed, for example, has not been heat-killed. In some embodiments, the bacterial strain in the composition of the invention has not been ated, for example, has not been heat-attenuated. For example, in some embodiments, the bacterial strain in the composition of the invention has not been killed, inactivated and/or attenuated. For example, in some embodiments, the bacterial strain in the composition of the invention is live. For example, in some embodiments, the bacterial strain in the composition of the invention is viable. For example, in some ments, the ial strain in the composition of the invention is capable of partially or totally sing the intestine. For example, in some embodiments, the bacterial strain in the composition of the invention is Viable and capable of partially or y colonising the intestine.

In some embodiments, the composition comprises a mixture of live bacterial s and bacterial strains that have been killed.

In preferred embodiments, the composition of the invention is encapsulated to enable ry of the bacterial strain to the intestine. Encapsulation protects the composition from degradation until delivery at the target location through, for e, rupturing with chemical or physical stimuli such as pressure, enzymatic ty, or physical disintegration, which may be triggered by changes in pH. Any appropriate encapsulation method may be used. Exemplary encapsulation techniques include entrapment within a porous matrix, attachment or adsorption on solid carrier surfaces, self-aggregation by flocculation or with cross—linking agents, and mechanical containment behind a microporous membrane or a microcapsule. Guidance on ulation that may be useful for preparing compcfims of the invention is available in, for example, references [41] and [42].

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm The composition may be administered orally and may be in the form of a tablet, capsule or powder.

Encapsulated products are preferred e Parabacteroides are bes. Other ingredients (such as vitamin C, for example), may be included as oxygen scavengers and prebiotic substrates to improve the delivery and / or partial or total colonisation and survival in viva. Alternatively, the probiotic composition of the invention may be administered orally as a food or nutritional product, such as milk or whey based fermented dairy product, or as a pharmaceutical product.

The composition may be formulated as a probiotic.

A composition of the invention es a therapeutically ive amount of a ial strain of the invention. A therapeutically effective amount of a bacterial strain is sufficient to exert a beneficial effect upon a patient. A therapeutically effective amount of a bacterial strain may be sufficient to result in delivery to and / or partial or total colonisation of the patient’s intestine.

A le daily dose of the bacteria, for e for an adult human, may be from about 1 x 103 to about 1 x 1011 colony forming units (CPU); for example, from about 1 x 107 to about 1 x 1010 CPU; in another example from about 1 x 106 to about 1 x 1010 CFU.

In certain embodiments, the composition contains the bacterial strain in an amount of from about 1 x 106 to about 1 x 1011 CFU/g, respect to the weight of the composition; for example, from about 1 x 108 to about 1 x 1010 CFU/g. The dose may be, for example, 1 g, 3g, 5g, and 10g.

Typically, a probiotic, such as the ition of the invention, is optionally combined with at least one suitable prebiotic compound. A prebiotic compound is usually a non-digestible carbohydrate such as an oligo- or polysaccharide, or a sugar alcohol, which is not degraded or absorbed in the upper digestive tract. Known prebiotics include commercial products such as inulin and transgalacto- oligosaccharides.

In certain embodiments, the probiotic composition of the present ion includes a prebiotic compound in an amount of from about 1 to about 30% by weight, respect to the total weight composition, (e.g. from 5 to 20% by weight). Carbohydrates may be selected from the group consisting of: fructo- accharides (or FOS), short-chain fructo-oligosaccharides, inulin, isomalt- oligosaccharides, pectins, xylo-oligosaccharides (or XOS), an—oligosaccharides (or COS), beta— glucans, arable gum modified and resistant starches, xtrose, D-tagatose, acacia fibers, carob, oats, and citrus fibers. In one aspect, the prebiotics are the short-chain fructo-oligosaccharides (for simplicity shown herein below as FOSs-c.c); said FOSs-c.c. are not ible ydrates, generally obtained by the conversion of the beet sugar and including a rose molecule to which three glucose molecules are bonded.

In certain ments, the compositions of the invention are used in combination with another therap compound for treating or preventing the neurodegenerative disorder. In some embodiments, the compositions of the invention are administered with nutritional supplements that [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm modulate neuroprotection or neuroproliferation. In preferred embodiments, the nutritional ments se or consist of nutritional vitamins. In certain embodiments, the vitamins are vitamin B6, ium, dimethylglycine (vitamin B16) and vitamin C. In certain embodiments, the compositions of the ion are administered in ation with another tic.

In certain embodiments, the compositions of the ion are for use in enhancing the effect of a second agent on a neurodegenerative disease. The immune modulatory effects of the compositions of the invention may make the brain more susceptible to conventional therapies such as Levodopa, ne agonists, MAO-B inhibitors, COMT inhibitors, Glutamate antagonists, or anticholinergics, which are exemplary secondary agents to be administered in combination (sequentially or contemporaneously) with the compositions of the invention.

The compositions of the invention may comprise pharmaceutically acceptable excipients or carriers.

Examples of such suitable excipients may be found in the reference [43]. Acceptable rs or diluents for therapeutic use are well known in the pharmaceutical art and are described, for example, in reference [44]. Examples of suitable carriers include lactose, starch, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol and the like. Examples of suitable diluents include ethanol, glycerol and water. The choice of pharmaceutical carrier, excipient or diluent can be selected with regard to the intended route of stration and standard pharmaceutical practice. The pharmaceutical compositions may comprise as, or in addition to, the carrier, excipient or diluent any suitable binder(s), lubricant(s), suspending agent(s), coating agent(s), solubilising agent(s). Examples of suitable binders include starch, gelatin, natural sugars such as glucose, anhydrous lactose, free—flow lactose, beta-lactose, corn sweeteners, natural and synthetic gums, such as acacia, tragacanth or sodium alginate, carboxymethyl cellulose and polyethylene glycol. Examples of le lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium te, sodium acetate, sodium chloride and the like. Preservatives, stabilizers, dyes and even flavouring agents may be provided in the pharmaceutical composition. Examples of preservatives include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. Antioxidants and ding agents may be also used.

The compositions of the invention may be formulated as a food product. For example, a food product may provide nutritional benefit in addition to the therapeutic effect of the invention, such as in a nutritional supplement. Similarly, a food t may be ated to enhance the taste of the ition of the invention or to make the composition more attractive to e by being more similar to a common food item, rather than to a pharmaceutical composition. In certain embodiments, the composition ofthe ion is formulated as a milk-based product. The term based product" means any liquid or semi-solid milk- or whey- based t having a varying fat content. The milk- based product can be, e.g., cow's milk, goat's milk, sheep's milk, d milk, whole milk, milk recombined from powdered milk and whey without any processing, or a processed product, such as urdled milk, curd, sour milk, sour whole milk, butter milk and other sour milk products.

Another important group includes milk beverages, such as whey beverages, fermented milks, [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm condensed milks, infant or baby milks; flavoured milks, ice cream; milk-containing food such as sweets.

In some embodiments, the compositions of the invention comprise one or more bacterial strains of the genus Parabacteroides and do not contain bacteria from any other genera, or which se only de s or biologically irrelevant amounts of ia from another genera. Thus, in some ments, the invention provides a composition comprising one or more bacterial strains of the genus Parabacteroides, which does not contain bacteria from any other genera or which ses only de minimis or biologically irrelevant amounts ofbacteria from another genera, for use in therapy.

In some embodiments, the compositions of the ion comprise one or more bacterial strains of the species Parabacteroides distasom's and do not contain bacteria from any other species, or which comprise only de minimis or biologically irrelevant amounts of bacteria from another species. Thus, in some embodiments, the invention provides a composition comprising one or more bacterial strains of the species Parabacteroides distasonis, which does not contain bacteria from any other species or which comprises only de minimis or biologically irrelevant amounts of bacteria from another species, for use in therapy.

In some embodiments, the compositions of the invention comprise one or more ial strains of the species Parabacteroides distasonis and do not contain ia from any other Parabacteroides species, or which comprise only de minimis or biologically irrelevant amounts of bacteria from r Parabacteroides species. Thus, in some embodiments, the invention provides a composition comprising one or more bacterial strains of the species Parabacteroides distasonz’s, which does not contain bacteria from any other Parabactemides species or which comprises only de minimis or ically irrelevant amounts of bacteria from another Parabacteroides species, for use in therapy.

In certain embodiments, the compositions of the invention contain a single bacterial strain or species and do not contain any other bacterial strains or species. Such compositions may comprise only de minimis or biologically irrelevant amounts of other bacterial strains or species. Such compositions may be a culture that is substantially free from other species of organism.

In some embodiments, the invention provides a composition comprising a single bacterial strain of the genus Parabacteroides, which does not contain bacteria from any other strains or which comprises only de minimis or ically vant s of bacteria from another strain for use in therapy.

In some embodiments, the invention provides a composition comprising a single bacterial strain of the species Parabacteroides distasonis, which does not contain bacteria from any other strains or which comprises only de minimis or biologically irrelevant s of bacteria from r strain for use in therapy.

In so bodiments, the compositions of the invention comprise more than one bacterial strain. For example, in some embodiments, the compositions of the invention comprise more than one strain from [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm within the same species (e.g. more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40 or 45 strains), and, optionally, do not contain bacteria from any other species. In some ments, the compositions of the invention comprise less than 50 strains from within the same species (e.g. less than 45, 40, 35, 30, 25, 20, 15, 12, 10, 9, 8, 7, 6, 5, 4 or 3 strains), and, optionally, do not contain bacteria from any other species. In some embodiments, the compositions of the invention se 1-40, 1-30, 1-20,1-19,1-18,1-15,1-10,1-9,1-8,1-7,1-6,1-5,1-4,1-3,1-2, 2-50, 2-40, 2-30, 2-20, 2-15, 2-10, 2-5, 6-30, 6-15, 16-25, or 31-50 strains from within the same species and, optionally, do not contain bacteria from any other species. The invention comprises any combination of the foregoing.

In some ments, the composition comprises a microbial consortium. For example, in some embodiments, the composition comprises the Parabacteroides ial strain as part of a microbial consortium. For example, in some embodiments, the cteroides bacterial strain is present in combination with one or more (e.g. at least 2, 3, 4, 5, 10, 15 or 20) other bacterial strains from other genera with which it can live symbiotically in vivo in the intestine. For example, in some embodiments, the composition comprises a ial strain of cteroides in combination with a bacterial strain from a ent genus. In some embodiments, the ial consortium comprises two or more bacterial strains obtained from a faeces sample of a single organism, e.g. a human. In some embodiments, the microbial consortium is not found together in nature. For example, in some ments, the ial consortium comprises bacterial strains obtained from faeces samples of at least two different organisms. In some ments, the two different organisms are from the same species, e.g. two different humans. In some embodiments, the two different organisms are an infant human and an adult human. In some embodiments, the two different organisms are a human and a non-human mammal.

In some embodiments, the composition of the invention additionally comprises a bacterial strain that has the same safety and therapeutic efficacy characteristics as strain MRXOOS, but which is not MRx0005, or which is not a cteroides distasonis.

In some embodiments in which the composition of the invention comprises more than one bacterial strain, species or genus, the individual bacterial strains, species or genera may be for separate, simultaneous or sequential administration. For example, the composition may comprise all of the more than one bacterial strain, s or genera, or the bacterial strains, species or genera may be stored separately and be administered separately, simultaneously or sequentially. In some embodiments, the more than one bacterial strains, species or genera are stored tely but are mixed together prior to use.

In some embodiments, the bacterial strain for use in the invention is obtained from human adult faeces.

In some embodiments in which the composition of the invention comprises more than one bacterial strain,fi)f the bacterial strains are ed from human adult faeces or if other bacterial strains are [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm present they are present only in de minimis amounts. The bacteria may have been cultured subsequent to being obtained from the human adult faeces and being used in a composition of the invention.

In some embodiments, the bacterial strain for use in the invention is obtained from human infant .

In some embodiments in which the composition of the invention comprises more than one bacterial strain, all of the ial s are obtained from human infant faeces or if other bacterial strains are t they are present only in de minimis amounts. The bacteria may have been cultured subsequent to being obtained from the human infant faeces and being used in a composition of the invention As mentioned above, in some embodiments, the one or more Parabacteroides bacterial strains is/are the only therapeutically active agent(s) in a composition of the invention. In some embodiments, the bacterial (s) in the composition is/are the only therapeutically active agent(s) in a composition of the invention.

The compositions for use in accordance with the invention may or may not require marketing approval.

In certain ments, the invention provides the above pharmaceutical composition, wherein said bacterial strain is lyophilised. In certain embodiments, the invention provides the above pharmaceutical composition, wherein said bacterial strain is spray dried. In certain embodiments, the invention provides the above pharmaceutical composition, wherein the bacterial strain is lyophilised or spray dried and wherein it is live. In certain embodiments, the invention provides the above pharmaceutical ition, n the bacterial strain is lyophilised or spray dried and wherein it is . In certain embodiments, the invention provides the above pharmaceutical ition, wherein the bacterial strain is lyophilised or spray dried and wherein it is capable of partially or totally sing the intestine. In certain embodiments, the invention provides the above pharmaceutical composition, wherein the ial strain is lyophilised or spray dried and wherein it is viable and capable of partially or totally sing the intestine.

In some cases, the lyophilised bacterial strain is reconstituted prior to administration. In some cases, the titution is by use of a diluent described herein.

The compositions of the invention can comprise pharmaceutically able excipients, diluents or carriers.

In n embodiments, the invention provides a pharmaceutical composition comprising: a bacterial strain of the invention; and a ceutically acceptable excipient, carrier or diluent; wherein the bacterial strain is in an amount sufficient to treat a neurodegenerative disorder when stered to a subject in need thereof.

In certain embodiments, the invention provides pharmaceutical composition comprising: a bacterial strain of the invention; and a pharmaceutically acceptable excipient, carrier or diluent; wherein the bacterurain is in an amount sufficient to treat or prevent a neurodegenerative disorder.

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm In certain embodiments, the invention provides the above pharmaceutical composition, wherein the amount of the bacterial strain is from about 1 X 103 to about l X 1011 colony forming units per gram with t to a weight of the composition.

In certain embodiments, the invention provides the above ceutical composition, wherein the composition is administered at a dose of l g, 3 g, 5 g or 10 g.

In certain ments, the invention provides the above pharmaceutical composition, wherein the composition is administered by a method selected from the group consisting of oral, , subcutaneous, nasal, , and sublingual.

In certain embodiments, the invention provides the above pharmaceutical composition, comprising a carrier selected from the group consisting of lactose, starch, e, methyl cellulose, magnesium stearate, ol and sorbitol.

In certain embodiments, the invention provides the above pharmaceutical composition, comprising a diluent selected from the group consisting of ethanol, glycerol and water.

In certain embodiments, the invention provides the above pharmaceutical composition, comprising an excipient selected from the group consisting of starch, gelatin, glucose, anhydrous e, free-flow lactose, beta-lactose, corn ner, acacia, tragacanth, sodium alginate, carboxymethyl cellulose, polyethylene glycol, sodium oleate, sodium stearate, magnesium te, sodium benzoate, sodium acetate and sodium chloride.

In n embodiments, the invention es the above pharmaceutical composition, further comprising at least one of a vative, an antioxidant and a stabilizer.

In certain embodiments, the invention provides the above ceutical composition, comprising a preservative selected from the group consisting of sodium benzoate, sorbic acid and esters of p— hydroxybenzoic acid.

In certain embodiments, the ion provides the above pharmaceutical composition, wherein said bacterial strain is lyophilised.

In certain embodiments, the invention provides the above pharmaceutical composition, wherein when the composition is stored in a sealed container at about 4°C or about 25°C and the ner is placed in an atmosphere having 50% relative humidity, at least 80% of the bacterial strain as measured in colony forming units, remains after a period of at least about: 1 month, 3 months, 6 months, 1 year, 1.5 years, 2 years, 2.5 years or 3 years.

In some embodiments, the composition of the invention is provided in a sealed container comprising a composition as described herein. In some embodiments, the sealed ner is a sachet or bottle. In some audiments, the composition ofthe invention is provided in a syringe comprising a composition as described herein.

[Annotation] kjm None set by kjm [Annotation] kjm ionNone set by kjm [Annotation] kjm ed set by kjm [Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm The composition of the present invention may, in some embodiments, be provided as a pharmaceutical formulation. For example, the ition may be provided as a tablet or capsule. In some embodiments, the capsule is a gelatine e cap").

In some embodiments, the compositions of the invention are administered . Oral administration may involve swallowing, so that the compound enters the intestinal tract, and/or buccal, lingual, or sublingual administration by which the compound enters the blood stream directly from the mouth.

Pharmaceutical formulations suitable for oral administration include solid plugs, solid microparticulates, semi-solid and liquid ding multiple phases or sed systems) such as tablets; soft or hard capsules containing multi- or nano-particulates, liquids (e.g. aqueous solutions), emulsions or powders; lozenges (including liquid-filled); chews; gels; fast dispersing dosage forms; films; ovules; sprays; and buccal/mucoadhesive patches.

In some embodiments the pharmaceutical formulation is an enteric formulation, i.e. a gastro-resistant formulation (for example, resistant to gastric pH) that is le for delivery of the composition of the invention to the intestine by oral administration. Enteric formulations may be particularly usefiil when the bacteria or another component of the composition is acid-sensitive, e.g. prone to degradation under gastric conditions.

In some embodiments, the enteric formulation ses an enteric g. In some embodiments, the formulation is an enteric-coated dosage form. For example, the formulation may be an enteric- coated tablet or an enteric-coated capsule, or the like. The enteric coating may be a conventional enteric coating, for example, a conventional coating for a tablet, capsule, or the like for oral delivery. The formulation may comprise a film coating, for example, a thin film layer of an enteric polymer, e.g. an acid-insoluble polymer.

In some embodiments, the enteric formulation is intrinsically enteric, for example, gastro-resistant t the need for an enteric coating. Thus, in some embodiments, the formulation is an c formulation that does not comprise an enteric coating. In some embodiments, the formulation is a capsule made from a thermogelling material. In some embodiments, the thermogelling material is a cellulosic material, such as methylcellulose, hydroxymethylcellulose or hydroxypropylmethylcellulose (HPMC). In some embodiments, the capsule comprises a shell that does not contain any film forming polymer. In some embodiments, the capsule comprises a shell and the shell comprises hydroxypropylmethylcellulose and does not comprise any film forming polymer (e.g. see [45 D. In some embodiments, the formulation is an intrinsically enteric capsule (for example, Vcaps® from Capsugel).

In some embodiments, the formulation is a soft capsule. Soft capsules are capsules which may, owing to addfiis of softeners, such as, for example, glycerol, ol, maltitol and polyethylene glycols, presen the capsule shell, have a certain elasticity and softness. Soft capsules can be produced, for example, on the basis of ne or starch. Gelatine-based soft capsules are commercially available [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm from various ers. ing on the method of administration, such as, for example, orally or rectally, soft capsules can have various shapes, they can be, for example, round, oval, oblong or torpedo-shaped. Soft capsules can be produced by conventional processes, such as, for example, by the Scherer s, the l process or the droplet or blowing process.

Culturing methods The bacterial strains for use in the present invention can be cultured using standard microbiology techniques as detailed in, for example, references [46-48].

The solid or liquid medium used for culture may be YCFA agar or YCFA medium. YCFA medium may include (per 100ml, approximate values): ne (1.0 g), yeast extract (0.25 g), NaHCO3 (0.4 g), cysteine (0.1 g), K2HPO4 (0.045 g), KH2P04 (0.045 g), NaCl (0.09 g), (NH4)2$O4 (0.09 g), MgSO4 ' 7HZO (0.009 g), CaClz (0.009 g), resazurin (0.1 mg), hemin (1 mg), biotin (1 ug), cobalamin (1 ug), p-aminobenzoic acid (3 ug), folic acid (5 ug), and pyridoxamine (15 ug).

Bacterial strainsfor use in e compositions The inventors have identified that the bacterial s of the ion are useful for treating or preventing neurodegenerative disorders. This is likely to be a result of the effect that the bacterial s of the ion have on the host immune system. Therefore, the compositions of the invention may also be useful for preventing neurodegenerative disorders, when administered as vaccine compositions. In certain such embodiments, the bacterial strains of the invention may be killed, inactivated or attenuated. In certain such embodiments, the compositions may comprise a vaccine adjuvant. In certain embodiments, the compositions are for administration via injection, such as via subcutaneous ion.

General The practice of the present invention will , unless otherwise indicated, conventional methods of chemistry, biochemistry, molecular y, logy and pharmacology, within the skill of the art. Such techniques are explained fully in the ture. See, e. g., references [49] and [50-56], etc.

The term "comprising" encompasses "including" as well as "consisting" e.g. a composition "comprising" X may t exclusively ofX or may include something additional e.g. X + Y.

The term "about" in relation to a numerical value x is optional and means, for example, x:10%.

The word "substantially" does not exclude "completely" e.g. a composition which is "substantially free" from Y may be completely free from Y. Where necessary, the word "substantially" may be omitted from the definition of the invention.

References to a percentage sequence identity n two nucleotide sequences means that, when alignegrat percentage of nucleotides are the same in comparing the two sequences. This alignmentand th cent homology or sequence identity can be determined using software programs known in ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm the art, for example those described in section 7.7.18 of ref. [57]. A preferred alignment is determined by the Smith-Waterman homology search algorithm using an affine gap search with a gap open penalty of 12 and a gap extension penalty of 2, BLOSUM matrix of 62. The Smith-Waterman homology search algorithm is disclosed in ref. [58].

Unless specifically stated, a process or method comprising numerous steps may comprise additional steps at the ing or end of the method, or may comprise additional intervening steps. Also, steps may be combined, omitted or med in an alternative order, if appropriate.

Various embodiments of the invention are described herein. It will be appreciated that the features specified in each embodiment may be combined with other ed features, to provide further embodiments. In particular, embodiments highlighted herein as being suitable, typical or preferred may be combined with each other (except when they are mutually exclusive).

MODES FOR CARRYING OUT THE INVENTION Example 1 — Efficacy erial inocala to act as a neuroprotectant Neuroblastoma cells were treated with compositions comprising ial strains according to the invention. The SH-SY5Y neuroblastoma cells used are ne producing and well-established as an in vitro model for studying neurodegenerative diseases. The ability of the bacterial strains to increase neuroproliferation was observed. The neuroblastoma cells were treated also treated dopaminergic neurotoxin ylpheny1pyridinium (MPP), which s permanent symptoms of Parkinson’s disease in neuroblastoma cells. The ability of the bacterial strains to act as a neuroprotectant against MPP was investigated.

Material and Methods Bacterial strain 755: Parabacteroides distasonis, haera massiliensis MRx0029 Cell line SH-SY5Y neuroblastoma cells were purchased from ECCACC (Cat. no: 94030304) and were grown in MEM (Sigma Aldrich, cat n. M2279) supplemented with Nutrient Mixture F-l2 Ham (Sigma h, cat 11. .

Method Once grown the SH—SYSY neuroblastoma cells were plated on 96-well plate at 11,000 cells/well and incubated for 2 days. The cells were then transferred to differentiation medium (which contains PBS at 1%fi10 uM retinoic acid (Sigma Aldrich, cat. n. R2625-100MG). entiation medium was replac ery other day and cells were harvested at 7 day of differentiation. Cells were pre-treated [Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm with or without MPP (Sigma h, cat. 11. G) for 8 hours. Subsequently, cells were treated with 10% bacterial supernatant and incubated overnight. Cell viability was ed by using CCK-8 reagent (Sigma Aldrich, Cell Counting Kit — 8, cat. n. 96992-3000TESTS-F) and read at 450nm wavelength.

Results The results of these experiments are shown in Figure 1. ent of neuroblastoma cells with MRx0005 or MRX0029 led to an increase in the proliferation of s. Neuroblastoma cells that were treated with MPP together with the bacterial strain had increased cell viability compared to the cells treated with MPP alone (which had decreased viability). These data show that the bacterial strains can act as a neuroprotectant.

Example 2A — Efficacy ofbacterial inocula to reduce IL-6 secretion.

Summary Activation of proinflammatory cytokines has been associated with neuron damage in neurodegenerative disease. Lipopolysaccharide (LPS) is a known ator of the proinflammatory cytokine IL-6. Human glioblastoma astrocytoma cells were treated with compositions comprising bacterial strains according to the invention in combination with LPS to e their ability to modulate the levels of IL-6.

Material and s Bacterial strain 755: cteroides distasom's Cell line MG U373 is a human glioblastoma astrocytoma derived from a malignant tumour and were purchased from Sigma-Aldrich (cat n. 08061901-1VL). MG U373 human glioblastoma astrocytoma cells were grown in MEM (Sigma Aldrich, cat n. M-2279) supplemented with 10% PBS, 1% Pen Strep, 4mM L-Glut, 1X MEM Non essential Amino Acid solution and 1X Sodium Piruvate.

Method Once grown the MG U373 cells were plated on 24—well plate at 100,000 cells/well. The cells were treated with LPS (lug/mL) alone or with 10% of bacteria supernatant from MRX0005 for 24h. A control was also med where the cells were incubated in untreated media. Afterwards the cell free supernatants were collected, centrifuged at 10,000g for 3min at 4°C. IL-6 was measured using the Human IL-6 ELISA Kit from Peprotech (cat n.#900-Kl6) ing to manufacturer instructions.

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Results The results of these experiments are shown in Figure 2A. Treatment of neuroblastoma cells with LPS and the bacteria strain led to a decrease in the level of IL-6 secreted.

Example ZB — cy ofbacterial inocula to modulate IL-8 secretion.

As neuro-inflammation plays a pivotal role in neurodegenerative diseases and IL-8 has been shown to have neuro-positive effects, the effect of itions comprising bacterial strains of the invention and LPS on the activation of lL-8 were assessed. Human glioblastoma ytoma cells were treated with compositions comprising bacterial strains according to the invention in combination with LPS to observe their ability to modulate the levels of lL-8.

Material and Methods Bacterial s Megasphaera massiliensis MRX0029; Parabacteroides distasonis MRX0005 Cell line MG U373 is a human glioblastoma astrocytoma derived from a malignant tumour and were purchased from Sigma-Aldrich (cat n. 08061901-1VL). MG U373 human glioblastoma astrocytoma cells were grown in MEM (Sigma Aldrich, cat n. M-2279) supplemented with 10% PBS, 1% Pen Strep, 4mM L-Glut, 1X MEM Non essential Amino Acid solution and 1X Sodium Piruvate.

Method Once grown the MG U373 cells were plated on 24-well plate at 100,000 well. The cells were treated with LPS L) alone or with 10% of bacteria supernatant from MRX0029 for 24h.

Afterwards the cell free atants were collected, centrifuged at 10,000g for 3min at 4°C. IL-8 was measured using Human lL-8 ELISA Kit from ech (cat n.#900-K18) according to manufacturer instruction.

Results The s of these experiments are shown in Figure 2B.

Example 2C — Efficacy ofbacterial inocula to reduce a-synaclein-induced inflammation.

Summary Neuroinflammation plays a pivotal role in Parkinson’s disease and a-synuclein has been shown to induce neuroinflammation in vivo. Therefore, the ability of the bacteria strains of the invention to inhibi ynuclein-induced neuroinflammation was assessed. A co-culture of human glioblastoma [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm astrocytoma cells and neuroblastoma cells were exposed to wild—type a-synuclein and the mutant isoforms E46K and A53T and treated with compositions comprising bacterial strains according to the invention. The ability of the bacteria strains to t a-synuclein-induced secretion of IL-6 was then tested.

Material and Methods Bacterial strains Megasphaera massiliensis MRX0029; Parabacteroides distasonis MRX0005 Cell line MG U373 is a human astoma astrocytoma derived from a malignant tumour and were purchased from Sigma-Aldrich (cat n. 08061901-1VL). MG U373 human glioblastoma astrocytoma cells were grown in MEM (Sigma Aldrich, cat n. M-2279) supplemented with 10% PBS, 1% Pen Strep, 4mM L-Glut, 1X MEM Non essential Amino Acid solution and 1X Sodium Piruvate.

SH—SY5Y is a human neurobastoma cell line derived from a malignant neuroblastoma and can be purchased from Aldrich (cat 11. 94030304-1VL). The cells were grown in 50 % MEM and 50% nt Mixture F-l2 Ham media supplemented with 2mM L-Glutamine, 10% heat inactivated FBS, 100 U/ml penicillin, 100 ug/ml streptomycin. Cells on growth medium were plated on 96-well plate at 11,000 cells/well and placed in the incubator. After 2 days, media were replaced with differentiation medium (growth medium ning 1% FBS) and 10 uM retinoic acid. Differentiation medium was replaced every other day and cells were used after 7 days of differentiation.

Method SHSY5Y cells were plated on 12 well plates at a y of 50,000 cells/well. The cells were grown in 50 % MEM and 50% Nutrient Mixture F-12 Ham media supplemented with 2mM L-Glutamine, 10% heat inactivated FBS, 100 U/ml penicillin, 100 ug/ml streptomycin. Cells on growth medium were plated on 96-well plate at 11,000 cells/well and placed in the incubator. After 2 days, media were replaced with entiation medium (growth medium ning 1% FB S) and 10 uM ic acid.

Differentiation medium was replaced every other day and cells were used after 7 days of differentiation. U373 were plated on 12 transwell plates (0.4um polyester membrane, Costar) at a y of 50,000cells/well for 72 hrs. Cells were co-cultured together for 24hrs before treatment in differentiation medium (growth medium containing 1% FBS without retinoic acid).

Thereafter cells were treated with 25rig/m1 clein (Wt, A53T, E46K) in the presence or e of 10% bacteria supernatant for 48 hrs. Cell free Supernatants were collected, spun-dwon at 10000g for 3 min at 4°C, aliquoted and stored at -80 0C. Human IL-6 and IL-8 were measured as described aboveu [Annotation] kjm None set by kjm [Annotation] kjm ionNone set by kjm ation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Results The s of these experiments are shown in Figure 3. Treatment of cells with wild-type a-synuclein and the mutant isoforms E46K and A53T induced moderate secretion of 1L-6. The a-syn-induced secretion of IL-6 was inhibited in cells treated with the bacteria strains.

Example 3 — Efficacy ofbacterial inocula to reduce NFKB activation Summary Activation of the NFKB promoter leads to the tion of proinflammatory cytokines ing IL- 18, IL-lcx, lL-18, TNFoc and IL-6. The NFKB promoter can be activated by oc-synuclein and LPS by stimulating the TLR4 ligand. Mutations in or—synuclein, such as uclein A53T, are implicated in familial Parkinson’s. Treatment of neuronal cells with LPS simulates Parkinson’s caused by environmental factors. The ability of itions comprising bacterial s ing to the invention to inhibit the activation of the NFKB promoter was igated.

Material and Methods Bacterial strain 755: Parabacteroides distasonis Cell line Human Hek blue TLR4 were purchased from InvivoGen (cat n. hkb-htlr4). Human Hek blue TLR4 were grown in DMEM high glucose (Sigma Aldrich, cat n. D-6171) supplemented with 10% FBS, 1% Pen Strep, 4mM L-Glut, Normocin and 1X HEK Blue selection solution.

Method Once grown the Human Hek blue cells were plated in 96 well plates at 25,000 cells/well in 4 replicates.

One set of cells were treated with oc-synuclein A53T (lug/mL) alone or with 10% of bacteria supernatant from MRx0005 for 22h. The second set of cells were treated with LPS (10 ng/mL, from Salmonella enterica serotype urium, Sigma Aldrich, cat n. L6143) alone or with 10% of bacteria supernatant from MRx0005 for 22h. The cells were subsequently spun down and 20ul of the supernatant was mixed with 200ul of Quanti Blue reagent (InvivoGen, cat n. rep-qb2), incubated for 2 h and absorbance read at 655nm.

Results The results of these experiments are shown in Figure 4 and 5. Figure 4 shows that the activation of the NFKB promoter by oc—synuclein is inhibited by MRx0005. Figure 5 shows that the activation of the NFKB promoter by LPS is inhibited by MRx0005.

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Example 4 — Efi‘icacy ofbacterial inocula to alter antioxidant capacity Summary The ability of compositions sing bacterial strains according to the invention to alter the antioxidant capacity. The antioxidant ty of the bacterial strain was established using the well- known ABTS (2,2'-azino-bis(3-ethylbenzothiazolinesulphonic acid)) assay.

Bacterial strain 755: cteroides distasonis Method Bacterial cells (106 or greater) were collected and centrifuged. They were resuspended in assay buffer (using three times the pellet volume). The suspension was sonicated on ice for 5 minutes and then spun down at 12,000 x g for 10 minutes. The supernatant was removed and measured using the ABTS assay kit produced by Sigma Aldrich (code ), in accordance with manufacturer’s instructions.

Results The results of these experiments are shown in Figure 6. Figure 6 shows that 5 has an idant capacity of approximately 0.5mM compared to .

Example 5 — Efiicacy ofbacterial inocula to alter lipidperoxidation levels Summary The ability of compositions comprising bacterial strains according to the ion to alter lipid peroxidation levels was investigated. The rbituric reactive substances assay (TBARs) was used to measure the by-products of lipid peroxidation.

Material and Methods Bacterial strain 755: Parabacteroides distasonis Method ial cells (106 or greater) were collected and centrifuged, a wash step was performed with isotonic saline before the pellet was re-suspensed in potassium chloride assay buffer. The suspension was sonicated on ice for 10 minutes and then spun down at 10,000 x g for 10 minutes. The supernatant was removed and the level of lipid peroxidation evaluated using the thiobarbituric ve substances assay.

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm ed set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Results The results of the experiments are shown in Figure 7. Figure 7 shows that MRx0005 is able to inhibit lipid dation by approximately 20 %, which is a higher antioxidant capacity than the positive control, butylated hydroxytoluene (1% w/v).

Example 6 — Efficacy ofbacterial inocula 0n histone deacetylatase activity Summary The ability of compositions comprising bacterial strains according to the invention to alter histone deacetylatase activity was investigated. Dysregulation of histone deacetylatase has been implicated in the pathogenesis associated with sociated neurodegenerative diseases.

Material and Methods Bacterial strain 755: Parabacteroides distasonis Cell line The cell line HT-29 was used e histone deacetylase is present.

Method Cell free supernatants ofstationary phase bacterial cultures were isolated by centrifugation and filtering in a 0.22 uM filter. HT-29 cells were used 3 days’ post confluence and stepped down in 1 mL DTS 24 hours prior to commencement of the experiment. The HT-29 cells were challenged with 10 % cell free supernatant diluted in DTS and this was left to te for 48 hours. se proteins were then extracted using the Sigma Aldrich Nuclease extraction kit and samples were snap frozen prior to HDAC activity measurement. HDAC activity was ed fluorometrically using the Sigma Aldrich (UK) kit.

The s of the experiments are shown in Figure 8. Figure 8 shows that MRx0005 is able reduce the levels of histone deacetylase activity.

Example 7 — Level ofindoleproduction in bacteria Summary The ability of the bacteria of the invention to produce indole was investigated. Indole has been implicated in attenuating inflammation and oxidative stress.

Material and Methods Bactez' Ijtrain 755: Parabacteroides distasonis ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ATCC 11775 is a bacterial reference strain that is known to produce .

Method Intact bacterial cells in stationary phase were incubated with 6mM phan for 48 hours. Bacterial species which possess the enzyme tryptophanase will utilise tryptophan as a substrate to produce indole. Following the 48 hour incubation period, the supernatant was removed and added to Kovac's reagent for fication of indole. Standards, stock solutions and reagents were prepared using rdised methods validated in-house.

Results The results of the experiments are shown in Figure 9. Figure 9 shows that 5 has the capacity to produce indole from tryptophan, at concentrations of approximately 0.2mM.

Example 8 — Level ofkynurenineproduction in bacteria Summary The ability of the bacteria of the ion to produce kynurenine was investigated. Dysregulation of the kynurenine pathway can lead to activation of the immune system and the accumulation of potentially neurotoxic compounds. Alterations in the kynurenine metabolism may be involved in the development of Parkinson’s diseases.

Bacterial strain 755: Parabacteroia’es distasonis DSM 17136 is a strain of Bacteroides copricola that is known to produce nine.

Method Cell free supernatants ofstationary phase bacterial cultures were isolated by centrifugation and filtering in a 0.22 uM filter and frozen until use. Kynurenine rds, stock solutions and reagents were prepared using standardised methods validated in—house. Sample were treated with trichloroacetic acid and centrifuged at 10,000xg for 10 minutes at 4°C. The supernatant was collected and sed into a 96 well plate. Ehrlich’s t was used for kynurenine detection and added at a ratio of 1:1.

Results The results of the experiments are shown in Figure 10. Figure 10 shows that MRx0005 has the capacity to produce kynurenine at a concentration of approximately 70 uM.

Example 9 — neuroprotection RA-differentiated SHSY-5Y cells were treated with MPP+, the active metabolite of MPTP, a chemical widelmeasdgas the rate of mitochondria respiration (Figured to mimic in vitro and in vivo some of the features of PD pathology. Cell Viability was 11). Both MRx0005 and MRx0029 showed [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm ation] kjm Unmarked set by kjm significant effects and promote per se an increase of the mitochondria metabolic activity in SHSY-SY cells. MRx0005 protection was about 20% compared to YCFA-MPP+ treated sample, about the same observed for the quercetin positive control (Fig. l 1).

Example 10 — Metaboliteproduction — metabolites in the brain Background Metabolites present in bacteria supernatants can directly influence the host response to oxidative , o-cell communication and neuroprotection. Metabolites that play a key role in neurological processes were measured during the ex vivo screening in brain tissue ofmice fed with MRx0005 and MRx0029.

Methods Animals BALBc (Envigo, UK) adult male mice were group housed under a 12 h light-dark cycle; standard rodent chow and water were available ad m. All experiments were performed in accordance with European guidelines following approval by University College Cork Animal Ethics Experimentation Committee. Animals were 8 weeks old at the start of the experiment.

Study Design Animals were allowed to habituate to their holding room for one week after arrival into the animal unit. They receive oral gavage (200uL dose) of live biotherapeutics at a dose of 1 X 109 CFU for 6 consecutive days between 15:00 and 17:00. On day 7, the s are decapitated, and tissues are harvested for experimentation.

Tissue Collection Animals were sacrificed in a random n regarding treatment and testing condition; ng occurred between 9.00 am. and 1:00 pm. Trunk blood was collected in potassium EDTA (Ethylene Diamine Tetra Acetic Acid) tubes and spun for 15 min at 4000 g. Plasma was isolated and stored at —80 0C for further analysis. The brain was quickly d, dissected and each brain region was snap— frozen on dry ice and stored at —80 °C for further analysis. Spleen was removed and processed immediately after culls for o immune stimulation. Intestinal tissue (2 cm segments of ileum and colon closest to the caecum were excised, and the furthest 1cm of tissue from the caecum were used) were mounted into the Ussing chambers for intestinal bility assay. The caecum was removed, weighted and stored at —80 °C for SCFAs analysis. ine is Neuronmitter concentration was analysed by HPLC on samples from the brainstem. Briefly, brainstem tissue was sonicated in 500 pl of chilled mobile phase spiked with 4 ng/40 ul of N—Methyl ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm -HT (Sigma Chemical Co., UK) as internal standard. The mobile phase contained 0.1 M citric acid, .6 mM octane-l-sulphonic acid (Sigma), 0.1 M sodium dihydrogen phosphate, 0.01 mM EDTA /Reagecon, Cork) and 9% (v/v) methanol (Alkem/Reagecon) and was adjusted to pH 2.8 using 4 N sodium hydroxide (Alkem/Reagecon). Homogenates were then centrifuged for 15 min at 22,000 X g at 4 oC and 40 ul of the supernatant injected onto the HPLC system which ted of a SCL 10- Avp system controller, LECD 6A electrochemical detector (Shimadzu), a LC-10AS pump, a CTO- 10A oven, a SIL-10A autoinjector (with sample cooler maintained at 40 C) and an online Gastorr Degasser (ISS, UK). A reverse-phase column (Kinetex 2.6 u C18 100 X 4.6 mm, Phenomenex) maintained at 30 0C was employed in the tion (Flow rate 0.9 mI/min). The glassy carbon working electrode combined with an Ag/AgCl reference electrode azu) operated a +0.8 V and the chromatograms generated were analyzed using Class-VP 5 software (Shimadzu). The neurotransmitters were fied by their characteristic retention times as determined by standard injections, which run at regular als during the sample is. The ratios of peak heights of analyte versus internal standard were measured and compared with rd injection. Results were expressed as ng of neurotransmitter per g fresh weight of tissue.

Metabolite analysis For GC-metabolite analysis, samples of bacterial supernatants were derivatized with methyl chloroformate using a slightly modified version of the protocol described by Smart et a1. (DOI: .103 8/nprot.2010.108). All samples were analyzed in a randomized order. Analysis was performed using GC (7890B, Agilent) coupled with a quadropole detector B, Agilent). The system was controlled by ChemStation (Agilent). Raw data was ted to netCDF format using Chemstation (Agilent), before the data was imported and processed in Matlab R2014b (Mathworks, Inc.) using the PARADISe software described by Johnsen et. al (DOI: 10.1016/j.chroma.2017.04.052).

For fatty acid analysis samples were acidified using hydrochloride acid, and deuterium labelled internal standards where added. All samples were analyzed in a randomized order. Analysis was performed using a high polarity column (ZebronTM ZB-FFAP, GC Cap. Column 30 m x 0.25 mm X 0.25 um) installed in a GC (7890B, Agilent) d with a quadropole detector (59977B, Agilent). The system was controlled by ChemStation (Agilent). Raw data was converted to netCDF format using Chemstation (Agilent), before the data was imported and processed in Matlab R2014b (Mathworks, Inc.) using the PARADISe software described by Johnsen et a1 (DOI: 10.1016/j.chroma.2017.04.052).

Results — neurotransmitter graduction The results are shown in Figure 12, which shows that in brains of mice fed with 9, noradrenaline levels are increased (p=0.0507), anied with a slight increase of serotonin and 5- HLAA. These data t the metabolite analysis set out below, suggesting that 29 is a major h 4-hydroxyphenylacetic acid, a known antioxidant (Weon et a1, 2016). More importantly, 4-hydroxyphenylacetic acid is a tic intermediate of dopamine and norepinephrine and an [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm important bio-active molecule (Huot et a1, Parkinson’s Disese 2015). In fact, in PD, degenerative changes extend beyond the nergic system, affecting equally the serotonergic and noradrenergic systems, which in turn leads to decreased levels of serotonin (5-hydroxytryptamine, 5-HT) and noradrenaline (norepinephrine) in both striatal and extra-striatal structures (Scatton B, Javoy-Agid F, Rouquier L, Dubois B, Agid Y Brain Res. 1983 Sep 26; 275(2):321-8.). L-DOPA targets mainly the dopamine-related features of PD, however it does not address the decreases in both 5-HT and noradrenaline. Adding to this is that the longer is the duration of L-DOPA ent, the more visible are a range of motor and nonmotor complications (e.g. esia, psychiatric symptoms) (Hely MA, Morris JG, Reid WG, ante R, Mov Disord. 2005 Feb; 20(2):190-9.) Therefore, these data demonstrate that bacteria that produce organic acids, such as 4-hydroxyphenylacetic acid or succinic acid, may be useful in therapy, in particular in the treatment of neurodegenerative diseases.

Results — metabolite production Metabolites present in bacteria supernatants can ly influence the host response to oxidative stress, cell-to—cell communication and neuroprotection in the specific. Metabolites in the atant of cultures of MRX0029 and MRX0005 were analysed and the results are shown in Figure 13.

A few metabolites showed a striking difference between the two strains analysed. The tration of succinic acid was particularly elevated in MRx0005. stingly, the ratio sample/media for 4- hydroxyphenylacetic acid was significantly higher in MRx0029 (Fig. 13).

Fatty acid is in the atants revealed an interesting dichotomy in the two strains: MRx0005 produced mainly acetic and propanoic acid, while MRx0029 produced butanoic, pentanoic and hexanoic acid, both in the linear and branched forms (Fig. 14B). The two strains looked very different and in particular, the production of succinic acid and 4-hydroxyphenylacetic acid by MRx0005 and MRx0029 respectively was notable (Figure 14A). Furthermore, MRx0005 seems to produce more C2 and C3 short chain fatty acids, while 29 produced more C4 ate) and both linear and branched medium chain fatty acids, including hexanoic acid. ic acid is a Krebs cycle metabolite involved in oxidative phosphorylation. Oxidative orylation complex is a key step for synaptic king of proteins and vesicles to proximal and distal regions (Budd SL and Nichols, 1998). Its dysfunction has been reported in neurodegenerative disorders including Alzheimer’s disease, Parkinson’s disease and Spinocerebellar ataxia type 1 (Manczak M et al. 2004; Ebadi et al, 2001). These findings are particularly interesting as succinic acid can augment mitochondrial activity and t vulnerable neurons in neurodegenerative disease related to misfolded proteins including PD (Ferroet al, Plos one, 2017). BDNF and succinic acid have both a similar protective activity not only in degeneration but also in mental disorders like depres'fi and anxiety, which are quite common amongst patients diagnosed with PD or AD.

[Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Figure 14B also demonstrates that MRX0029 is a butyrate (butanoic acid) producer. This may be significant because butyrate has a known role is reducing impermeability of the blood brain barrier, which has a neuroprotective effect [59]. This property of MRx0029 (and other neuroprotective bacteria) may contribute to its efficacy.

Example 11 - Modulation ofthe mRNA expression oftightjunction proteins Since recent evidence ts that intestinal dysfunction and inflammation is a non-motor symptom associated with PD, the ability of the bacterial strains of the invention to cause any intestinal r dysfunction was investigated. HT29-mtx epithelial, mucin—producing cell monolayers (Gagnon et al, J Microbiological Methods, 2013) were used as an in vitro model to evaluate gut barrier tion and immune stimulation following treatment with MRx0005 and MRx0029. Differentiated HT29-mtx cells exposed to phorbol 12-myristateacetate (PMA) secreted a significant amount of IL-8; in contrast treatment for 24h with MRx005 and MRx0029 bacterial supernatants, induced an even lower ion of IL-8 compared than both untreated and YCFA-treated cells (Fig. 14A).

The ability of MRx0005 and MRx0029 to regulate epithelial permeability by modifying intracellular signal transduction ed in the sion and localization of proteins involved in the gut r formation was then investigated.

RNA was isolated and Quantitative RT-PCR (qRT-PCR) analysis was performed to characterize the changes in gene expression of tight junction proteins during tion with MRx0005 and 9.

The stration of MRx0029 enhanced Occludin, Vlillin, Tight on Protein 1 and 2 (respectively TJPl and TJP2) mRNA expression after 2h incubation (Fig. 14B). In contrast, exposure to MRx0005 did not alter the gene expression of tight junction proteins indicating that the two strains act differentially on the intestinal barrier.

The in vitro results were compared with data from the ex vivo parallel analysis on the gut of mice fed with MRx0005 and MRx0029. Gene expression of TJP2 and occludin was fied in the colon and ileum. The ex vivo data perfectly mirror the in vitro data as MRx0029 was able to significantly up- regulate TJPl and Occludin (p=0.073) in the colon region of the murine intestine (Fig. 14C+14D).

MRx0029 was also able to decrease the bility function in the colon of the same mice (Fig. 14E+14F).

Materials and s - RNA extraction and qPCR is Total RNA was extracted using the RNeasy mini kit (Qiagen, Manchester, JUK) according to the manufacturer's instructions, and the RNA concentration determined by absorbance at 260/280 nm using Dectrophotometer (nano-Drop ND-1000; Thermo Scientific, Wilmington, DE). For mRNA express1on analysis, cDNA was prepared from 2000 ng of total RNA using the High-Capacity cDNA ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm reverse transcription kit (Thermo Fisher, Loughborough) according to the manufacturer's instructions.

The reverse ription reactions were performed in a thermo cycler (Biometra, y) at 25°C for 10 min, 37°C for 120 min, and 85°C for 5 min. Resulting cDNA was amplified in duplicates by the SYBR-Green PCR assay, and products were detected on QuantStudio 7 real-time PCR machine (Applied tems, UK) using a standardised profile (initial denaturation of 95°C for 10 minutes, followed by 40 cycles of 10 seconds of denaturation at 95°C and 30 seconds of annealing/extension at 60°C). A dissociation stage was added after the 40 cycles to generate a g curve. Analysis was performed using the Applied Biosystems QuantStudio Real-Time PCR Software V1.2. The primer sequences for Actin, Villin, Occludin TJPl and TJP2 are ed in the sequence listing.

Example 12 — Level ofBDNF secretion in SHSY-5Y cells Background Brain-derived neurotrophic factor (BDNF) is a ubiquitous molecule in the brain associated with neural development, neuro—protection and neuro-regeneration. BDNF not only ts against egeneration but also mental disorders like depression and anxiety, which are quite common amongst patients diagnosed with PD or AD.

SH—SY5-SY were plated in 24 wells plate at density of 60,000 cells/well and placed in the tor.

After 24 h, media were replaced with differentiation medium (growth medium containing 1% FBS) and 10 uM retinoic acid. Differentiation medium was replaced every other day and cells were used on day 10 of differentiation. For the treatment differentiation medium was removed and ed with 450ul of full growth media and 50 ul of bacteria SN was added to the treated wells or YCFA+ was added as ve Control.

Results The results are shown in Figure 15, which shows that administration ofMRX0005 in combination with retinoic acid increases the secretion of BDNF from differentiated neuroblastoma cells.

Example 13 — Efficacy ofbacterial inocula to reduce oxidative levels in cells Background The generation of reactive oxygen species contributes to the pathology of neurodegenerative diseases.

The ability of bacterial s to protect differentiated SHSY-SY and U373 cells from reactive oxygen species (ROS) generated by treatment with Tert-Butyl Hydrogen Peroxide (TBHP) was investigated.

Material and Methods Bactegd'train Megasplzaera massiliensis MRX0029 [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Method Y cells were plated in black flat bottom 96 well plate at density of 5000 cells/well and placed in the CO2 incubator. After 24 h, media were replaced with differentiation medium (growth medium containing 1% FB S) and 10 iiM ic acid. Differentiation medium was replaced every other day.

On Day 10 the differentiation medium was removed and cells were washed with rmed PBS and stained with 10uM DCFDA molecular probe for 20 mins in growth medium ning 1% FBS. Then cells were washed with rmed PBS again and treated with 100uM TBHP in the presence or absence of 10% bacteria supernatant for 2h. Fluorescence ity was measured using TECAN plate reader at Ex/Em 485/530 nm.

Results The results of the experiments are shown in Figure 16. Figure 16b shows that MRX0005 is able to inhibit ROS production in differentiated SHSY-SY neuroblastoma cells. MRX0005 also reduces the generation of ROS in astroglioblastoma cells (Figure 16a). This shows that MRX0005 has general antioxidant activity.

Example 14 — Stability testing A composition described herein containing at least one bacterial strain described herein is stored in a sealed container at 25°C or 4°C and the container is placed in an atmosphere having 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90% or 95% relative humidity. After 1 month, 2 months, 3 months, 6 , 1 year, 1.5 years, 2 years, 2.5 years or 3 years, at least 50%, 60%, 70%, 80% or 90% of the bacterial strain shall remain as measured in colony forming units determined by standard protocols. e 15 Animals The animals and study design used were the same as for Example 10.

Bacterial strains 0 755: Parabacteroides distasonis (MRX005) 0 Megasphaera massiliensis (MRX0029) Tissue Collection Animals were sacrificed in a random n regarding treatment and testing condition; sampling occurred between 9.00 am. and 2:30 pm. Trunk blood was collected in potassium EDTA (Ethylene Diamine Tetra Acetic Acid) tubes and spun for 15 min at 4000 g. Plasma was isolated and stored at —80 OCD further analysis. The brain was quickly excised, dissected and each brain region was snap- frozen on dry ice and stored at —80 0C for further analysis. Spleen was removed, collected in 5 mL [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm RPMI media (with L-glutamine and sodium bicarbonate, R8758 Sigma + 10 % FBS , Sigma) + 1% Pen/Strep (P4333, Sigma)) and sed immediately after culls for ex-vivo immune stimulation. Intestinal tissue (2 3cm ts of ileum and colon closest to the caecum were excised, and the furthest 1cm 2cm oftissue from the caecum were used) were d into the Ussing rs for intestinal permeability assay. The caecum was removed, weighted and stored at -80 °C for SCFAs analysis.

Monoamine Analysis The neurotransmitter concentration was analysed as described in Example 10 Spleen Cytokine Assay Spleens were collected immediately in 5mL RPMI media following sacrifice and cultured immediately. Spleen cells were first homogenised in this RPMI media, followed by 5 mins incubation with lml of RBC lysis buffer (11814389001 ROCHE, Sigma). A further 10 ml of RPMI media was added, followed by 200G centrifugation for 5 mins. The atant was then filtered through 40um strainer. Cells were counted and seeded (4,000,000/mL . After 2.5 h of adaptation, cells were stimulated with lipopolysaccharide (LPS-2 ug/ml) or concanavalin A (ConA-2.5 ug/ml) for 24 h.

Following stimulation, the supernatants were harvested to assess the cytokine e using Proinflammatory Panel 1 (mouse) V-PLEX Kit (Meso Scale Discovery, Maryland, USA) for TNFOt, IL-10, IL-lB, Interferon y, CXCL2 and IL6. The analyses were performed using MESO QuickPlex SQ 120, SECTOR Imager 2400, SECTOR Imager 6000, SECTOR S 600.

Gene Expression Analysis Total RNA was extracted using the mirVanaTM miRNA Isolation kit (Ambion/Llife technologies, Paisley, UK) and DNase treated (Turbo DNA-free, /life technologies) according to the manufacturers recommendations. RNA was quantified using NanoDropTM spectrophotometer (Thermo Fisher Scientific Inc., gton, Delaware, USA) according to the manufacturer's instructions. RNA quality was assessed using the Agilent Bioanalyzer (Agilent, Stockport, UK) according to the manufacturer's procedure and an RNA integrity number (RIN) was calculated. RNA with RIN value >7 was used for subsequent experiments. RNA was reverse transcribed to cDNA using the Applied Biosystems High Capacity cDNA kit (Applied Biosystems, Warrington, UK) according to manufacturer's ctions. Briefly, Multiscribe Reverse riptase (50 U/uL) (1)(2)(1)(10) was added as part of RT master mix, incubated for 25°C for 10 min, 37°C for 2 h, 85°C for 5 min and stored at 4°C. Quantitative PCR was carried out using probes (6 carboxy fluorescein - FAM) designed by Applied Biosystems to mouse specific targeted genes, while using B—actin as an endogenous control.

Amplification ons contained 1 ul cDNA, 5 ul of the 2X PCR Master mix ), 900 nM of each fir and were brought to a total of 10 ul by the addition of RNase-free water. All reactions were rmed in triplicate using 96-well plates on the LightCycler®480 System. Thermal cycling ions were as recommended by the cturer (Roche) for 55 cycles. To check for amplicon [Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ination, each run contained no template controls in triplicate for each probe used. Cycle threshold (Ct) values were recorded. Data was normalized using B-actin and transformed using the 2-AACT method and presented as a fold change vs. control group.

Short Chain Fatty Acids is in the Caecal Content Caecum content was mixed and vortexed with MilliQ water and incubated at room ature for 10 min. Supernatants were obtained by centrifugation (10000 g, 5 min, 4 °C) to pellet bacteria and other solids and filtration by 0.2um. It was transferred to a clear GC vial and 2-Ethylbutyric acid (Sigma) was used as the internal standard. The concentration of SCFA was analyzed using a Varian 3500 GC flame-ionization system, fitted with a with a ZB-FFAP column (30 m x 0.32 mm x 0.25 mm; Phenomenex). A standard curve was built with different concentrations of a standard mix ning acetate, nate, iso-butyrate, n—butyrate, isovalerate and valerate (Sigma). Peaks were integrated by using the Varian Star Chromatography ation version 6.0 software. All SCFA data are expressed as umol/g.

Statistical Analysis Normally distributed data are presented as mean i SEM; Non-parametric datasets are presented as median with inter-quartile range. Unpaired iled t-test were applied to analyse parametric data and Mann-Whitney test was used for non-parametric. Spearman's rank correlation coefficient was employed for the correlation analysis in the pooled datasets. A p value < 0.05 was deemed significant in all cases. s — Neurotransmitter production The results in Figure 17 show the effect 05 treatment on the concentration of neurotransmitters in the brain of mice. Most y, treatment with MRx005 leads to a se in dopamine.

Results — Gene expression Expression of genes for neurotransmitter receptors [serotonin receptor 1a(5-HTR1a), ne D1 receptor, GABA or subunit Bl, GABAA receptor, NMDA2A (Grin2A) and NMDA2B (Grin2b) receptor], atory markers [IL-lB, 1L6, CDllb, TNFor and TLR4], and endocrine markers [corticosterone releasing factor (CRF), corticosterone releasing factor receptors 1 and 2 (CRFRl, CRFR2), brain-derived neurotrophin factor (BDNF), vasopressin receptor, oxytocin receptor, glucocorticoid receptor and mineralocorticoid receptor] were analysed in brain tissue from the hippocampus, amygdala and prefrontal cortex.

Figures 18-32 show the changes in gene expression after MRX005 or MRX0029 treatment in the hippocampal, amygdala and prefrontal . Treatment with MRx0029 led to an increase in gluCOCficoid receptor expression in the amygdala (Figure 25C). Figure 26A shows that MRx005 [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm significantly increased the expression of BDNF in the amygdala, while treatment with MRx0029 significantly increased the expression of TLR4 in the amygdala (Figure 26B).

Both MRx005 and MRx0029 can increase sion of CD1 1b in the la (Figure 27A), while the expression of IL-6, Grin2a and Grin2b is reduced after MRx005 ent (Figures 27B-D). In addition, MRx005 and MRx0029 significantly sed the expression of GABRA2 and increased the expression of GABBRl in the amygdala.

Treatment with MRx005 led to a significant increase in the sion ofBDNF in the prefrontal cortex (Figure 29B).

Discussion MRx005 and MRx0029 administration caused changes in gene sion in the amygdala. , especially Results — Effect on Tphl and [DO-I expression Figure 33 shows that MRx0029 can significantly increase the expression tryptophan ylase- 1 (Tphl) in the colon and that MRX005 treatment can increase lDO-l sion in the colon. Treatment with MRX005 increased the expression of Tphl and IDOl in the ileum, while MRX029 had no effect on the expression of these genes in the ileum (Figure 34).

Indoleamine-pyrrole 2,3-dioxygenase-l (IDO-l) the first and imiting enzyme in the tryptophan/kynurenine y while tryptophan hydroxylase l (Tphl), an isoform of the enzyme tryptophan hydroxylase, responsible for the synthesis of serotonin. These data suggest that MRx0029 and MRx005 may affect serotonin levels and the tryptophan/kynurenine pathway.

Results — Effect on phan metabolite levels Figure 35 shows the effect of treatment with MRx005 on the levels of circulating kynurenine and tryptophan. s — Effect on cvtokine expressionflom splenocytes The ex-vivo splenocyte assay involves challenging the splenocytes (cells isolated from the spleen - a main organ involved in immune defence), with a bacterio- or viral-mimetic challenge.

MRX005 significantly reduced the levels of interferon-y in splenocytes following a nge with LPS (Figure 36). In addition, MRX005 reduced the levels of interleukin-6 and tumour necrosis factor after a challenge with LPS (Figures 38 and 39, respectively). Treatment with MRx0029 led to a reduction in interferon-y, interleukin-18 and interleukin-6 following a challenge with LPS (Figures 36, 37 and 38, respectively).

Treatment with MRx005 and MRx0029 led to an increase in the levels of the chemoattractant CXCLl (Figuru).

[Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm s — Effect on Caecal Snort Chain Fatty Acid Levels Short chain fatty acids (SCFAs) are produced when non-digestible fibres from the diet are fermented by ia in the gut. The effects of MRXOOS administration are shown in Figure 42.

Example I 6 — Farther analysis ofMRX029 and MRX005 changes in gene expression levels Cell line SH—SY5Y cells Bacterial strains 0 755: Parabacteroides distasonis (MRX005) 0 Megasphaera massilz'ensis (MRX0029) qPCR SH-SY5Y were plated in 10cm petri dishes a density of 2x106 cells. After 24h cells were treated in differentiation medium (growth medium containing 1% FBS without RA) with 10% bacteria supernatants or YCFA+, 10uM RA, 200uM hexanoic acid or 200uM valproic acid, for 17 hrs. There after representative images were taken using phase contrast EVOS XL core microscope at 40X/0.65 magnification. Cells were collected, and total RNA was ed ing to RNeasy mini kit protocol (Qiagen). cDNAs were made using the high capacity cDNA reverse transcription kit (Applied Biosystems). Gene expression was measured using qPCR. GAPDH was used as internal control. Fold change was calculated according to the 20AM) . The primer sequences for MAP2, DRD2, GABRB3, SYP, PlNKl, PARK7 and NSE are provided in the sequence listing.

Immuno-labelling and cell imaging Cells were seeded onto 8-well r slides (Marienfeld Laboratory Glassware) at 5x104 cells/well overnight and were treated with 10% bacterial supernatant for 24 h. For entiation, cells were treated with 10 nM RA for 5 days before ng with cell-free ial atant for 24 h.

Afterwards, the cells were fixed with 4% paraformaldehyde in PBS for 20 minutes at room temperature (RT). Fixed cells were washed with PBS, and permeabilized with 1% Triton X-100 in PBS for 10 minutes. After washing with PBS, the slides were incubated with blocking buffer (4% BSA/PBS) for 1 h at RT before adding anti-MAP2 antibody or BS-tubulin 421 and sc-80005 respectively, Santa Cruz Biotechnology Inc) diluted in 1% BSA/PBS for 12 h at 4°C. They were then washed twice with PBS, followed by incubation with Alexa Flour 488 conjugated ouse (Molecular Probes Inc) and Alexa Flour 594 conjugated Phalloidin (ab176757, Abcam) for 1 h at RT. After washing 3X with PBS, the slides were staining with DAPI and mounted with Vectashield® (Vector Laboratories). Slides were viewe ng a Axioskop 50 microscope (Zeiss) equipped with a 63x/1.2 W Korr objective and filter sets suitable for detection of the fluorochromes used. Manual exposure times for the digital acquisition [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm of images immuno-labelled with MAP-2 were kept constant allowing comparison between different wells and treatments. Phalloidin in) and DAPI re times varied to suit the field of view.

Randomised fields of view were acquired using a QImaging camera controlled by Image Pro Plus software. Images were saved as TIFF files and opened in Adobe Photoshop CC 2015 .l.2. Images of the MAP-2, DAPI and Phalloidin images were then overlaid and merged. Representative images were selected to illustrate the differences in abundance and location of the proteins examined.

Immunoblotting SH-SY5Y cells cultured under the indicated conditions described above, treated with MRx0005 and MRx0029 for 24h and then lysed in RIPA buffer containing cocktail of protease inhibitors (Roche Diagnostics, UK). n concentration was estimated using the BCA protein assay kit e Biotechnology, Rockford, IL), separated by GE and transferred to a PVDF membrane.

Membranes were then blocked with 5% t dry milk or 5% BSA and incubated overnight at 4°C with the primary antibodies ctively MAP2 and B3-tubulin). The blots were then incubated with the riate horseradish peroxidase (HRP)-conjugated secondary antibody, and proteins were detected by chemiluminescence detection kit (Pierce Biotechnology, Rockford, IL). For both MAP2 and B3-tubulin, B-actin served as a l to monitor protein loading variability amongst s.

Results and Discussion Gene exgression Figure 43 shows the MRx0029 and MRXOOS—induced changes in expression levels of Actin, Villin, Occludin TJPl, TJP2, MAP2, DRD2, GABRB3, SYP, PINKl, PARK7 and NSE.

Resuits — Microscopy and Immunoblotting Figure 44 shows the change in the level of expression of MAP2 in SHSY5Y cells as determined by confocal microscopy. The expression levels of MAP2 and B3-tubulin were also quantified by immunoblot analysis. The results shown in Figure 44M and N indicate that MRX029 induces an increase in the level expression of MAP2 Sequences Additional primers used in qPCR (with SEQ ID NO in brackets) Gene ID Forward sequence e sequence NSE CCCTGTATCGTAAGAACGGT (30) GCCACCATTGATCACGTTGA (31) [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Pl NK1 GGCAGCACATCAGGGTAGTC CCCAAGCAACTAGCCCCTC (32) (33) PARK7 GTAGCCGTGATGTGGTCATTT (34) CTGTGCGCCCAGATTACCT (35) SYP CTCGGCTTTGTGAAGGTGCT GGCTTCATGGCATCAACTTCA (36) (37) Sequences SEQ ID NO:1 (Parabacteroides onis gene for 16S mal RNA, l ce, strain: JCM 5825 - AB238922) agagtttgat cctggctcag gatgaacgct agcgacaggc ttaacacatg caagtcgagg i ggcagcgggg tgtagcaata caccgccggc gaccggcgca cgggtgagta atgc Z aacttgccta tcagaggggg ataacccggc gaaagtcgga ctaataccgc atgaagcagg gcat gggaatattt gctaaagatt catcgctgat agataggcat gcgttccatt 1 aggcagttgg cggggtaacg gcccaccaaa atgg ataggggttc tgagaggaag gtcccccaca ctga gacacggacc ctac gggaggcagc agtgaggaat Z attggtcaat gggcgtaagc ctgaaccagc caagtcgcgt gagggatgaa atgg atcgtaaacc tcttttataa gggaataaag tgcgggacgt gtcccgtttt gtatgtacct i tatgaataag gatcggctaa ctccgtgcca gcagccgcgg taatacggag gatccgagcg Z ttatccggat ttattgggtt taaagggtgc gtaggcggcc ttttaagtca gcggtgaaag tctgtggctc aaccatagaa ttgccgttga aactgggggg cttgagtatg tttgaggcag 1 gcggaatgcg tggtgtagcg gtgaaatgca tagatatcac gcagaacccc gattgcgaag gcagcctgcc aagccattac tgacgctgat gcacgaaagc gtggggatca aacaggatta Z gataccctgg tagtccacgc agtaaacgat gatcactagc tgtttgcgat acactgtaag Z cggcacagcg aaagcgttaa gtgatccacc tggggagtac gccggcaacg ctca i aaggaattga cgggggcccg cacaagcgga ggaacatgtg gtttaattcg atgatacgcg Z aggaacctta tttg aacgcattcg gaccgaggtg gaaacacctt ttctagcaat agccgtttgc gaggtgctgc atggttgtcg tcagctcgtg ccgtgaggtg tcggcttaag Z tgccataacg agcgcaaccc ttgccactag ttactaacag gttaggctga ggactctggt gggactgcca gctg cgaggaaggc ggggatgacg tcaaatcagc acggccctta Z catccggggc gacacacgtg ttacaatggc gtggacaaag ggaggccacc tggcgacagg Z gagcgaatcc ccaaaccacg tctcagttcg gatcggagtc tgcaacccga ctccgtgaag i ctggattcgc tagtaatcgc gcatcagcca tggcgcggtg aatacgttcc cgggccttgt Z acacaccgcc cgtcaagcca tgggagccgg gggtacctga agtccgtaac cgaaaggatc 144; ggcctagggt aaaactggtg actggggcta agtcgtaaca aggtaacc SEQ ID N022 (Parabacteroz'des distasonis gene for 16S ribosomal RNA, partial sequence, : JCM 13400 - 23) D agagtttgat tcag gatgaacgct agcgacaggc ttaacacatg caagtcgagg 61 ggcagcacag gtagcaatac gcga ccggcgcacg ggtgagtaac gcgtatgcaa ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Z cttacctatc agagggggat aacccggcga gact aataccgcat gaagcagggg Z ccccgcatgg ggatatttgc taaagattca tcgctgatag ataggcatgc gttccattag ggcg gggtaacggc ccaccaaacc gacgatggat aggggttctg agaggaaggt Z cccccacatt ggtactgaga cacggaccaa actcctacgg gcag atat tggtcaatgg gcgtaagcct gcca agtcgcgtga gggatgaagg ttctatggat Z cgtaaacctc ttttataagg gaataaagtg cgggacgtgt cctgttttgt atgtacctta tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga cgtt Z atccggattt attgggttta aagggtgcgt aggcggcctt ttaagtcagc ggtgaaagtc Z tgtggctcaa ccatagaatt gccgttgaaa ctggggggct tgagtatgtt tgaggcaggc ggaatgcgtg gtgtagcggt gaaatgctta gatatcacgc agaaccccga ttgcgaaggc Z agcctgccaa gccatgactg acgctgatgc acgaaagcgt ggggatcaaa caggattaga ggta gtccacgcag taaacgatga gctg tttgcgatac agtgtaagcg Z gcacagcgaa agcgttaagt gatccacctg gggagtacgc cggcaacggt gaaactcaaa gacg ggggcccgca caagcggagg aacatgtggt ttaattcgat gatacgcgag Z gaaccttacc cgggtttgaa cgcattcgga ccgaggtgga aacacctttt ctagcaatag Z ccgtttgcga ggtgctgcat ggttgtcgtc agctcgtgcc gtgaggtgtc ggcttaagtg ccataacgag cctt gccactagtt actaacaggt gagg actctggtgg Z gactgccagc gtaagctgcg aggaaggcgg ggatgacgtc aaatcagcac ggcccttaca tccggggcga cacacgtgtt acaatggcgt ggacaaaggg aagccacctg gcgacaggga Z gcgaatcccc aaaccacgtc tcagttcgga tcggagtctg caacccgact ccgtgaagct ggattcgcta gtaatcgcgc atcagccatg gcgcggtgaa tacgttcccg ggccttgtac i acaccgcccg tcaagccatg ggagccgggg gtacctgaag tccgtaaccg tcgg Z cctagggtaa aactggtgac tggggctaag tcgtaacaag gtaacc SEQ ID N023 (Parabacteroides distasonis gene for 16S mal RNA, partial sequence, strain: JCM 13401 - AB238924) 1 agagtttgat cctggctcag gatgaacgct agcgacaggc ttaacacatg caagtcgagg 6; ggcagcacag gtagcaatac ccgccggcga ccggcgcacg ggtgagtaac gcgtatgcaa l2; cttgcctatc agagggggat aacccggcga aagtcggact aataccgcat gaagcagggg 18; ccccgcatgg ggatatttgc taaagattca tcgctgatag ataggcatgc gttccattag 24; gcagttggcg gggtaacggc aacc gacgatggat aggggttctg agaggaaggt - cccccacatt ggtactgaga cacggaccaa actcctacgg gcag tgaggaatat 36; tggtcaatgg gcgtaagcct gaaccagcca agtcgcgtga gggatgaagg ttctatggat 42; cgtaaacctc ttttataagg gaataaagtg tgggacgtgt ttgt atgtacctta 48; tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga cgtt 54; atccggattt attgggttta aagggtgcgt aggcggcctt ttaagtcagc ggtgaaagtc 60; tcaa ccatagaatt gccgttgaaa ctgggaggct tgagtatgtt tgaggcaggc 66; ggaatgcgtg gtgtagcggt gaaatgctta gatatcacgc agaaccccga ttgcgaaggc 72- ccaa gccatgactg acgctgatgc acgaaagcgt ggggatcaaa caggattaga 40 78; taccctggta gcag taaacgatga tcactagctg tttgcgatac agcg fii gcacagcgaa agcgttaagt gatccacctg gggagtacgc cggcaacggt gaaactcaaa ggaattgacg ggggcccgca gagg tggt ttaattcgat gatacgcgag 96; gaaccttacc tgaa cgcattcgga ccgaggtgga aacacctttt ctagcaatag [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm 1021 ccgtttgcga ggtgctgcat ggttgtcgtc agctcgtgcc gtgaggtgtc ggcttaagtg 1081 cgag cgcaaccctt gccactagtt actaacaggt gatgctgagg actctggtgg 1141 cagc gtaagctgcg aggaaggcgg ggatgacgtc aaatcagcac ggcccttaca 1201 tccggggcga cacacgtgtt acaatggcgt ggacaaaggg atgccacctg ggga 1261 gcgaatcccc aaaccacgtc cgga tcggagtctg caacccgact ccgtgaagct 1321 ggattcgcta gtaatcgcgc atcagccatg tgaa tacgttcccg ggccttgtac 1381 acaccgcccg tcaagccatg ggagccgggg gaag tccgtaaccg aaaggatcgg 1441 cctagggtaa aactggtgac tggggctaag tcgtaacaag gtaacc SEQ ID NO:4 (Parabacteroides distasom's gene for 16S ribosomal RNA, partial sequence, strain: JCM 13402 - 25) 1 agagtttgat cctggctcag cgct agcgacaggc ttaacacatg caagtcgagg ggcagcacag gtagcaatac cgggtggcga ccggcgcacg ggtgagtaac gcgtatgcaa Z cttacctatc agagggggat aacccggcga aagtcggact aataccgcat gaagcagggg ccccgcatgg ggatatttgc taaagattca tcgctgatag ataggcatgc gttccattag 1 gcagttggcg gggtaacggc aacc gacgatggat aggggttctg agaggaaggt Z cccccacatt ggtactgaga cacggaccaa actcctacgg gaggcagcag tgaggaatat i tggtcaatgg gcgtaagcct gaaccagcca agtcgcgtga gggatgaagg ttctatggat Z cgtaaacctc ttttataagg gaataaagtg cgggacgtgt cccgttttgt atgtacctta tgaataagga aact ccgtgccagc agccgcggta agga tccgagcgtt Z atccggattt attgggttta aagggtgcgt aggcggcctt ttaagtcagc ggtgaaagtc tgtggctcaa ccatagaatt gccgttgaaa ctgggaggct tgagtatgtt tgaggcaggc 1 ggaatgcgtg gtgtagcggt gaaatgctta acgc agaaccccga ttgcgaaggc Z agcctgccaa gccatgactg acgctgatgc acgaaagcgt ggggatcaaa caggattaga taccctggta gtccacgcag taaacgatga tcactagctg tttgcgatac actgtaagcg Z gcacagcgaa agcgttaagt gatccacctg gggagtacgc cggcaacggt gaaactcaaa ggaattgacg ggggcccgca caagcggagg aacatgtggt ttaattcgat cgag Z gaaccttacc cgggtttgaa cgcattcgga ccgaggtgga aacacctttt ctagcaatag ccgtttgcga ggtgctgcat ggttgtcgtc agctcgtgcc gtgaggtgtc ggcttaagtg Z cgag cgcaaccctt gccactagtt actaacaggt aaagctgagg actctggtgg Z gactgccagc gtaagctgcg aggaaggcgg ggatgacgtc aaatcagcac ggcccttaca tccggggcga cacacgtgtt acaatggcgt aggg aggccacctg gcgacaggga Z cccc aaaccacgtc tcagttcgga tcggagtctg gact ccgtgaagct gcta gcgc atcagccatg gcgcggtgaa tacgttcccg gtac Z acaccgcccg tcaagccatg ggagccgggg gtacctgaag tccgtaaccg aaaggatcgg cctagggtaa aactggtgac taag tcgtaacaag gtaacc SEQ ID N025 (Parabacteroides distasonis gene for 16S ribosomal RNA, l sequence, strain: JCM 13403 - AB23 8926) 40 D1 agagtttgat cctggctcag gatgaacgct agcgacaggc ttaacacatg caagtcgagg 61 ggcagcacag gtagcaatac cgggtggcga ccggcgcacg ggtgagtaac gcgtatgcaa ation] kjm None set by kjm [Annotation] kjm ionNone set by kjm [Annotation] kjm ed set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm 1 cttacctatc agagggggat aacccggcga aagtcggact aataccgcat gaagcagggg Z ccccgcatgg ggatatttgc taaagattca tcgctgatag ataggcatgc gttccattag gcagttggcg gggtaacggc ccaccaaacc gacgatggat aggggttctg agaggaaggt Z cccccacatt ggtactgaga cacggaccaa actcctacgg gaggcagcag tgaggaatat atgg gcgtaagcct gaaccagcca gtga gggatgaagg ttctatggat Z cgtaaacctc ttttataagg gaataaagtg tgggacgtgt ttgt atgtacctta tgaataagga tcggctaact ccgtgccagc ggta atacggagga tccgagcgtt Z atccggattt attgggttta gcgt aggcggcctt ttaagtcagc ggtgaaagtc Z tgtggctcaa ccatagaatt gccgttgaaa ctgggaggct tgagtatgtt tgaggcaggc ggaatgcgtg gtgtagcggt gaaatgctta gatatcacgc agaaccccga ttgcgaaggc Z agcctgccaa gccatgactg acgctgatgc acgaaagcgt ggggatcaaa caggattaga taccctggta gtccacgcag taaacgatga tcactagctg tttgcgatac attgtaagcg Z gcacagcgaa agcgttaagt gatccacctg gggagtacgc cggcaacggt caaa ggaattgacg ggggcccgca caagcggagg aacatgtggt ttaattcgat gatacgcgag Z gaaccttacc cgggtttgaa cgcattcgga ccgaggtgga aacacctttt ctagcaatag Z ccgtttgcga ggtgctgcat ggttgtcgtc agctcgtgcc tgtc ggcttaagtg ccataacgag cgcaaccctt gccactagtt actaacaggt aaagctgagg actctggtgg Z gactgccagc gtaagctgcg aggaaggcgg ggatgacgtc aaatcagcac ggcccttaca gcga cacacgtgtt acaatggcgt ggacaaaggg aggccacctg gcgacaggga Z gcgaatcccc aaaccacgtc tcagttcgga tcggagtctg caacccgact ccgtgaagct ggattcgcta gcgc atcagccatg gcgcggtgaa tacgttcccg ggccttgtac i acaccgcccg catg ggagccgggg gtacctgaag tccgtaaccg aaaggatcgg Z cctagggtaa aactggtgac tggggctaag tcgtaacaag gtaacc SEQ ID N026 (Parabacteroides distasonis gene for 16S ribosomal RNA, l sequence, strain: JCM 13404 - AB23 8927) agagtttgat cctggctcag gatgaacgct agcgacaggc ttaacacatg caagtcgagg Z ggcagcacag atac cgggtggcga ccggcgcacg ggtgagtaac gcgtatgcaa cttacctatc agagggggat aacccggcga aagtcggact aataccgcat gaagcagggg Z ccccgcatgg ggatatttgc taaagattca atag ataggcatgc ttag 1 ggcg gggtaacggc ccaccaaacc gacgatggat aggggttctg agaggaaggt cccccacatt gaga cacggaccaa actcctacgg gaggcagcag tgaggaatat Z tggtcaatgg gcct gaaccagcca agtcgcgtga gggatgaagg ttctatggat cgtaaacctc ttttataagg gaataaagtg tgggacgtgt cccgttttgt atgtacctta Z tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga tccgagcgtt atccggattt attgggttta aagggtgcgt cctt ttaagtcagc ggtgaaagtc Z tgtggctcaa ccatagaatt gccgttgaaa ctgggaggct tgagtatgtt tgaggcaggc 1 ggaatgcgtg gtgtagcggt gaaatgctta gatatcacgc agaaccccga ttgcgaaggc agcctgccaa gccatgactg acgctgatgc acgaaagcgt ggggatcaaa caggattaga 40 Z taccctggta gtccacgcag taaacgatga tcactagctg atac attgtaagcg gcacagcgaa agcgttaagt gatccacctg gggagtacgc cggcaacggt gaaactcaaa Z ggaattgacg ggggcccgca caagcggagg tggt ttaattcgat gatacgcgag gaaccttacc cgggtttgaa cgcattcgga ccgaggtgga aacacctttt ctagcaatag ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm 1021 ccgtttgcga ggtgctgcat ggttgtcgtc agctcgtgcc gtgaggtgtc agtg 1081 ccataacgag cgcaaccctt gccactagtt actaacaggt aaagctgagg actctggtgg 1141 gactgccagc gtaagctgcg aggaaggcgg ggatgacgtc aaatcagcac taca 1201 tccggggcga cacacgtgtt acaatggcgt ggacaaaggg aggccacctg gcgacaggga 1261 cccc aaaccacgtc tcagttcgga tctg gact ccgtgaagct 1321 ggattcgcta gtaatcgcgc atcagccatg gcgcggtgaa cccg ggccttgtac 1381 acaccgcccg tcaagccatg ggagccgggg gtacctgaag tccgtaaccg aaaggatcgg 1441 cctagggtaa aactggtgac taag tcgtaacaag gtaacc SEQ ID NO:7 (Parabacteroides merdae gene for 16S ribosomal RNA, partial sequence, strain: JCM 9497 - AB238928) Z tgat cctggctcag gatgaacgct agcgacaggc ttaacacatg caagtcgagg atga tttgtagcaa tacagattga tggcgaccgg cgcacgggtg gcgt Z atgcaactta agag ggggatagcc cggcgaaagt cggattaata ccccataaaa caggggtccc gcatgggaat atttgttaaa gattcatcgc tgatagatag gcatgcgttc Z cattaggcag ttggcggggt aacggcccac caaaccgacg atggataggg gttctgagag Z gaaggtcccc cacattggta ctgagacacg gaccaaactc ctacgggagg cagcagtgag i gaatattggt ccga gaggctgaac agtc gcgtgaagga agaaggatct Z atggtttgta aacttctttt ataggggaat aaagtggagg acgtgtcctt ttttgtatgt accctatgaa taagcatcgg ctaactccgt gccagcagcc gcggtaatac ggaggatgcg Z agcgttatcc ggatttattg ggtttaaagg gtgcgtaggt ggtgatttaa gtcagcggtg aaagtttgtg gctcaaccat aaaattgccg ttgaaactgg gttacttgag tgtgtttgag 1 gtaggcggaa tgcgtggtgt agcggtgaaa tgcatagata tcacgcagaa ctccgattgc Z gaaggcagct acca taactgacac tgaagcacga aagcgtgggg atcaaacagg attagatacc ctggtagtcc acgcagtaaa cgatgattac tttg cgatacaatg Z taagctctac agcgaaagcg ttaagtaatc cacctgggga gtacgccggc aacggtgaaa ctcaaaggaa ttgacggggg cccgcacaag cggaggaaca tgtggtttaa ttcgatgata Z gaac cttacccggg tttgaacgta gtctgaccgg agtggaaaca ctccttctag caatagcaga ttacgaggtg ctgcatggtt gtcgtcagct cgtgccgtga ggtgtcggct Z taagtgccat aacgagcgca acccttatca acta acaggtgaag ctgaggactc Z tggtgagact gccagcgtaa gctgtgagga aggtggggat gacgtcaaat cagcacggcc cttacatccg gggcgacaca acaa tggcatggac aaagggcagc tacctggcga Z gcta atctccaaac catgtctcag ttcggatcgg agtctgcaac tcgactccgt gaagctggat tcgctagtaa tcgcgcatca gccatggcgc ggtgaatacg ttcccgggcc Z acac cgcccgtcaa gccatgggag ccgggggtac ctgaagtccg taaccgcaag gatcggccta gggtaaaact ggtgactggg gctaagtcgt aacaaggtaa CC SEQ ID NO:8 (Parabacteroides merdae gene for 16S ribosomal RNA, partial sequence, strain: JCM 13405 - AB23 8929) 40 D1 agagtttgat cctggctcag gatgaacgct agcgacaggc ttaacacatg gagg 61 ggcagcatga tttgtagcaa tacagattga tggcgaccgg cgcacgggtg agtaacgcgt [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm ed set by kjm 121 atgcaactta agag agcc cggcgaaagt cggattaata ccccataaaa 18; caggggttcc gcatgggaat atttgttaaa gattcatcgc tgatagatag gcatgcgttc 24- cattaggcag gggt aacggcccac caaaccgacg atggataggg gttctgagag 301 gaaggtcccc cacattggta ctgagacacg gaccaaactc ctacgggagg cagcagtgag 36L gaatattggt caatggccga gaggctgaac cagccaagtc gcgtgaagga atct 42L atggtttgta tttt ataggggaat aaagtggagg acgtgtcctt ttttgtatgt 48; accctatgaa taagcatcgg ctaactccgt gccagcagcc gcggtaatac ggaggatgcg 54L atcc ggatttattg ggtttaaagg gtgcgtaggt ggtgatttaa gtcagcggtg 60; aaagtttgtg gctcaaccat aaaattgccg ttgaaactgg gttacttgag tgtgtttgag 66- ggaa tgcgtggtgt agcggtgaaa tgcatagata tcacgcagaa ctccgattgc 72L gaaggcagct acca acac tgaagcacga aagcgtgggg atcaaacagg 78L tacc ctggtagtcc acgcagtaaa cgatgattac taggagtttg cgatacaatg 841 taagctctac agcgaaagcg ttaagtaatc cacctgggga gtacgccggc aacggtgaaa 90; ctcaaaggaa ttgacggggg cccgcacaag cggaggaaca ttaa ttcgatgata 96L cgcgaggaac cttacccggg tttgaacgta gtctgaccgg aaca ctccttctag 102; caatagcaga ggtg ctgcatggtt gtcgtcagct cgtgccgtga ggtgtcggct 108- ccat aacgagcgca acccttatca ctagttacta acaggtgaag ctgaggactc 1142 tggtgagact gccagcgtaa gctgtgagga aggtggggat gacgtcaaat cagcacggcc 120- cttacatccg gggcgacaca cgtgttacaa tggcatggac aaagggcagc tacctggcga 1262 caggatgcta atctccaaac catgtctcag ttcggatcgg agtctgcaac tcgactccgt 132; gaagctggat tcgctagtaa tcgcgcatca gccatggcgc ggtgaatacg ttcccgggcc 1381 ttgtacacac cgcccgtcaa gccatgggag ccgggggtac ctgaagtccg taaccgcaag 144C gatcggccta gggtaaaact ggtgactggg gctaagtcgt aacaaggtaa cc SEQ ID N019 (consensus 16S rRNA sequence for Parabacteroides distasonis strain 755) AMCCGGGTGGCGACCGGCGCACGGGTGAGTAACGCGTATGCAACTTGCCTATCAGAGGGGGATAACCCGGCGAAAGT CGGACTAATACCGCATGAAGCAGGGATCCCGCATGGGAATATTTGCTAAAGATTCATCGCTGATAGATAGGCATGCG TAGGCAGTTGGCGGGGTAACGGCCCACCAAACCGACGATGGATAGGGGTTCTGAGAGGAAGGTCCCCCACA TTGGTACTGAGACACGGACCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGGCGTGAGCCTGAACC AGCCAAGTCGCGTGAGGGATGAAGGTTCTATGGATCGTAAACCTCTTTTATAAGGGAATAAAGTGCGGGACGTGTCC CGTTTTGTATGTACCTTATGAATAAGGATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATCCGAGCGT TATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCGGCCTT7TAAGTCAGCGGTGAAAGTCTGTGGCTCAACCATAG AATTGCCGTTGAAACTGGGAGGCTTGAGTATGTTTGAGGCAGGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGAT ATCACGCAGAACCCCGATTGCGAAGGCAGCCTGCCAAGCCATTACTGACGCTGATGCACGAAAGCGTGGGGATCAAA CAGGATTAGATACCCTGGTAGTCCACGCAGTAAACGATGATCACTAGCTGTTTGCGATACACTGTAAGCGGCACAGC GAAAGCGTTAAGTGATCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAG CGGAGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGTTTGAACGCATTCGGACMGAKGTGGAA ACACATTTTCTAGCAATAGCCATTTGCGAGGTGCTGCATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCGGCTTAAG TGCCATAACGAGCGCAACCCTTGCCACTAGTTACTAACAGGTAAAGCTGAGGACTCTGGTGGGACTGCCAGCGTAAG 40 CTGCGAGGAAGGCGGGGATGACGTCAAATCAGCACGGCCCTTACATCCGGGGCGACACACGTGTTACAATGGCGTGG GGAAGCCACCTGGCGACAGGGAGCGAATCCCCAAACCACGTCTCAGTTCGGATCGGAGTCTGCAACCCGAC TCCGTGAAGCTGGATTCGCTAGTAATCGCGCATCAGCCATGGCGCGGTGAATACGTTCCCGGGCCTTGTACACACCG CCCGTCAAGCCATGGGAGCCGGGGGTACCTGAAGTCCGTAACCGCGAGGATCGGCCTAGGGTAAAACTGGTGACTGG GGCTAAGTCGTACGGGG SEQ ID NO:10 (strain 755 genome sequence) — see electronic sequence listing.

SEQ IRON nsus 16S rRNA sequence for Megasphaera massiliensis strain MRX0029) [Annotation] kjm None set by kjm ation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm ed set by kjm TGAGAAGCTTGCTTCTTATCGATTCTAGTGGCAAACGGGTGAGTAACGCGTAAGCAACCTGCCCTTCAGATGGGGAC AACAGCTGGAAACGGCTGCTAATACCGAATACGTTCTTTCCGCCGCATGACGGGAAGAAGAAAGGGAGGCCTTCGGG CTTTCGCTGGAGGAGGGGCTTGCGTCTGATTAGCTAGTTGGAGGGGTAACGGCCCACCAAGGCGACGATCAGTAGCC GGTCTGAGAGGATGAACGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATCTT CCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAACGATGACGGCCTTCGGGTTGTAAAGTTCTGTTATATG GGACGAACAGGACATCGGTTAATACCCGGTGTCTTTGACGGTACCGTAAGAGAAAGCCACGGCTAACTACGTGCCAG CAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCGCGCAGGCGGCATCGCAAGT CGGTCTTAAAAGTGCGGGGCTTAACCCCGTGAGGGGACCGAAACTGTGAAGCTCGAGTGTCGGAGAGGAAAGCGGAA TTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAAGCGGCTTTCTGGACGACAACTGA CGCTGAGGCGCGAAAGCCAGGGGAGCAAACGGGATTAGATACCCCGGTAGTCCTGGCCGTAAACGATGGATACTAGG TGTAGGAGGTATCGACTCCTTCTGTGCCGGAGTTAACGCAATAAGTATCCCGCCTGGGGAGTACGGCCGCAAGGCTG AAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGTATGTGGTTTAATTCGACGCAACGCGAAGAACCTTA CTTGACATTGATTGCTACGGAAAGAGATTTCCGGTTCTTCTTCGGAAGACAAGAAAACAGGTGGTGCACGG CTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATCTTCTGTTGCCAGCACC TCGGGTGGGGACTCAGAAGAGACTGCCGCAGACAATGCGGAGGAAGGCGGGGATGACGTCAAGTCATCATGCCCCTT ATGGCTTGGGCTACACACGTACTACAATGGCTCTTAATAGAGGGAAGCGAAGGAGCGATCCGGAGCAAACCCCAAAA ACAGAGTCCCAGTTCGGATTGCAGGCTGCAACTCGCCTGCATGAAGCAGGAATCGCTAGTAATCGCAGGTCAGCATA CTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAAAGTCATTCACACCCGAAGCCGGTGA GGCAACCGCAAG Primers used for qPCR (with SEQ ID NO in brackets) Forward sequence [fieverse sequence GAICAAGATCATTGQICCTC((12) gETTGTC§AGAAA§GGTGT§§C (1%) GTGGAAGGACTCATG (l4) ATGCCAGTGAGCTTCCCGTTC (15) CTCAGCACCGCTAACAGAGG (l6) EECATTGGCGCTTCTCTCCTC (17) AAGAGGAATTTTGACACTGG (l8) GCCATGTACTCTTCACTTTC (19) AAGTCACACTGGTGAAATCC (20) ;;CTCTTGCTGCCAAACTATCT (21) CCCTCCCCTGGATCAGGAT (22) GCCATCAAACTCGTCCATCA (23) $95111 CTGCTCTACGTTTG (24) [AGATGGACATAAGATGAGGTG (25) REFERENCES Spor et a], (2011) Nat Rev Microbiol. 79-90.

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VVC)2013/050792 \V /046580 \V 13/008039 ation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm [Annotation] kjm None set by kjm [Annotation] kjm MigrationNone set by kjm [Annotation] kjm Unmarked set by kjm Goldin and Gorbach (2008) Clin Infect Dis. 46 Suppl 2:S96-100.

Azad et al. (2013) BMJ. 347zf6471.

Mayer et al (2014) The Journal ofNeuroscience 34(46):15490 415496 Cryan and Dinan (2015) Neuropsychopharmacology, 40: 241-2.

Zhou and Foster (2015) Neuropsychiatric Disease and Treatment 11: 715—723.

Wang and Kasper (2014) Brain Behav Immun. 38: 1412.

Chapter 38 - Nonsteroidal anti-inflammatory drugs exposure and the central nervous system (2014) Handbook ofClinical Neurology 119, 577-584 Sakamoto and Benno (2006) Int JSyst Evol Microbiol. 56(Pt 7):1599-605.

Masco et al. (2003) Systematic andApplied Microbiology, —563.

Srfitkova et al. (2011) J. iol. Methods, 87(1):10—6.

Pal R et a1, Neurol Res 2016, 38(12): 1 1 1 1-1 122 Daniele SG et al, Sci Signal 2015, 8(376):ra45 Wang et al. (2016) JNeurogastroenterol Motil 22: 589—605.

Foguem & Manckoundia (2018) Current Neurology and Neuroscience Reports, 18: 24 Ludolph et al. (2009) Eur JNeurol. 16(3): 29743 09.

Galpern & Lang (2006) Neurological Progress 59 (3) 449-458 Zadori et al (2012) Journal ofNeural Transmission, 119, 2, 2754283 Lee et al (2008) European J. Cell Biology 873894397 Pirooznia and Elefant (2013) Front Cell Neurosci. 7: 30.

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Wang et al. (2015) PNAS 112(9):2583-2858 Psaty et al. (2003) JAMA, 289(19):2534—44 Lancet. (1995) 346(899148992):1647453 [3 8] Miyamoto—Shinohara et al. (2008).]. Gen. Appl. Microbiol, 54, 9424.

Cryopreservation and Freeze—Drying Protocols, ed. by Day and McLellan, Humana Press.

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Print Out (Original in Electronic Form) (This sheet is not part of and does not count as a sheet of the international application) 0-1 Form PCT/R011 34 Indications Relating to Deposited Microorganism(s) or Other Biological Material (PCT Rule 13bis) 01 Prepared Using PCT Online Filing Version 0.256e MT/FOP 20141031/0 .20.5 .20 0-2 ational Application No. 0-3 Applicant's or agent's tile reierence P0 7 0 7 7 1W0 The Indications made below relate to the deposited microorganism(s) or other biological al ed to In the description on: 1-1 page 8 1-2 line 1-7 1-3 ldentitication of deposit 11 Name °'dep°s"arvi"5‘"""°" NCIMB National Collections of Industrial, Food and Marine Bacteria (NCIMB) 1—3-2 Address°fdep°snaryins‘i‘u‘im NCIMB Ltd, Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen A321 9YA, United Kingdom 13 Dateofdeposii 12 March 2015 (12.03.2015) 14 Accession Number NCIMB 42382 1-5 Designated States for Which All designations Indications are Made The indications made below relate to the ted microorganism(s) or other biological material reierred to in the ption on: 2-1 Page 11 2-2 Ime 22-27 2-3 Identification of deposit 21 Name °'deP°5"a’Vi"5‘"""°" NCIMB National Collections of Industrial, Food and Marine Bacteria (NCIMB) 22 s°fdep°5"a'y‘nsmu‘im NCIMB Ltd, Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen A321 9YA, United Kingdom 23 dep°5" 13 July 2017 (13.07.2017) 2—3-4 Accession Number NCIMB 42787 2-5 Designated States Ior Which All designations Indications are Made FOR RECEIVING OFFICE USE ONLY 0-4 This form was received with the international application: yes (yes or no) 01 Authorized officer Desplanque, Ludlvme. .

Print Out nal in Electronic Form) (This sheet is not part of and does not count as a sheet of the international application) FOR INTERNATIONAL BUREAU USE ONLY 0-5 This torm was received by the international Bureau on: 01 Authorized officer

Claims (17)

1. Use of a composition comprising a bacterial strain of the genus Parabacteroides , in the manufacture of a medicament for ng or ting a neurodegenerative er selected from the group consisting of Parkinson’s disease, including progressive supranuclear palsy, progressive 5 supranuclear palsy, Steele-Richardson-Olszewski syndrome, normal pressure hydrocephalus, ar or arteriosclerotic parkinsonism and drug-induced parkinsonism; mer’s disease, including Benson's me; Huntington’s disease; amyotrophic lateral sclerosis; Lou Gehrig's disease; motor neurone disease; prion disease; spinocerebellar ataxia; spinal muscular atrophy; dementia, including Lewy body, vascular and frontotemporal dementia; primary progressive 10 aphasia; mild cognitive impairment; HIV-related ive impairment and corticobasal degeneration in a patient.

2. The use of claim 2, wherein the medicament is for treating or preventing Parkinson’s disease.

3. The use of claim 1 or claim 2, wherein the ment is for treating or preventing earlyonset egenerative disease.

4. The use of any one of the preceding claims, n the medicament is for preventing or 20 delaying onset or progression of a neurodegenerative disorder.

5. Use of a composition comprising a bacterial strain of the genus Parabacteroides , in the manufacture of a ment for treating brain injury in a patient. 25

6. The use of claim 5, wherein the brain injury is stroke, such as cerebral ischemia, focal cerebral ischemia, ischemic stroke or hemorrhagic stroke.

7. The use of any one of the preceding claims, wherein the bacterial strain is of Parabacteroides distasonis.

8. The use of any one of the preceding claims, wherein the bacterial strain has a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to the 16s rRNA sequence of a bacterial strain of Parabacteroides distasonis.

9. The use of any one of claims 1-7, wherein the bacterial strain has a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to SEQ ID NO:1, 2, 3, 4, 5, 6, 7 8 or 9. 5

10. The use of claim 9, wherein the ial strain has a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% cal to SEQ ID NO:9, or wherein the bacterial strain has the 16s rRNA ce represented by SEQ ID NO:9.

11. The use of claim 1 or claim 2, wherein the medicament comprises a bacterial strain of the 10 s Parabacteroides distasonis.

12. The use of any one of the preceding claims, wherein the medicament is formulated for oral administration. 15

13. The use of any one of the preceding claims, wherein the composition comprises one or more pharmaceutically acceptable excipients or carriers.

14. The use of any one of the preceding claims, wherein the bacterial strain is lyophilised. 20

15. Use of a food product comprising the composition comprising a bacterial strain of the genus

16.Parabacteroides , in the manufacture of a medicament for treating or preventing a neurodegenerative er in a patient, wherein the egenerative disorder is not multiple sclerosis. 25 16. The use of claim 15, modified by the features of any one of claims 1-14.

17. The use of any one of claims 1-4, wherein the bacterial strain is a cell of the Parabacteroides distasonis strain deposited under accession number NCIMB 42382.