Compositions from Aronia melanocarpa

Abstract

The present invention relates to compositions for use in treating cardiovascular disease (CVD) and / or gut dysbiosis and methods of treating CVD and / or gut dysbiosis. In particular, the present invention relates to compositions and methods for use to treat cardiovascular disease (CVD) and / or gut dysbiosis in subjects with reduced gut gene counts (fecal microbiome gene counts).

Description

[Technical field]

[0001] FIELD OF THEINVENTION The present invention relates to compositions for use in treating and / or preventing cardiovascular disease (CVD) and / or gut dysbiosis, and methods of treating CVD and / or gut dysbiosis. In particular, the present invention relates to compositions and methods for use to treat cardiovascular disease (CVD) and / or gut dysbiosis in subjects with reduced gut gene counts (fecal microbiome gene counts). [Background technology]

[0002] 2. Background of the Invention CVD, also referred to as heart disease, is a general term used to refer to conditions that affect the heart or circulation and includes coronary artery disease, stroke, and hypertension. CVD is the number one cause of death worldwide, accounting for an estimated 31% of all deaths worldwide (World Health Organization 2017). Each year, CVD accounts for almost 27% of deaths in the UK, and the total healthcare costs associated with CVD in the UK are estimated at £9 billion per year (British Heart Foundation).

[0003] Atherosclerosis refers to a progressive disease characterized by the accumulation of lipid and fibrous plaque in the arteries, leading to hardening and narrowing of these large blood vessels. Atherosclerosis occurs in the intima, the innermost of the three layers of the artery.

[0004] Cardiovascular events can occur due to the accumulation of atherosclerotic plaques that block or narrow the arterial lumen. Atherosclerosis-based CVD is one of the major causes of vascular disease worldwide. Indeed, after a slow progression throughout life, the disease can eventually lead to peripheral vascular disease and / or stroke in the older general population. Several risk factors, such as hypercholesterolemia, hypertension, smoking, and diabetes, have been implicated in the development and progression of atherosclerosis.

[0005] Blood pressure (BP) has been shown to be a robust surrogate marker of CVD risk, with a 3 mmHg reduction in systolic blood pressure (SBP) associated with a 5% reduction in the risk of CVD mortality.

[0006] Although the mechanism behind the association between BP and atherosclerosis has not yet been fully identified, the importance of the endothelium and the process of oxidative stress occurring at its level is considered to be the main explanation. However, while NO plays a key role in regulating vascular tone and blood pressure, it has also been observed that impaired NO activity is an essential component in the development of hypertension. BP pulsatile factors have been shown to induce atheroma instability and ultimately plaque rupture, indicating a mediating role of BP on atherosclerosis.

[0007] The accumulation of atherosclerotic plaque leads to hardening of the arterial walls. Pulse wave velocity (PWV) is the most used technique to assess arterial stiffness. PWV measures the speed of pressure waves traveling through the arterial system using applanation tonometry technique. PWV is expressed in m / s and is calculated based on dividing the distance between two measurement points by the transit time of the pulse pressure wave at these two locations. In a common method, the carotid and femoral arteries are the preferred sites for the assessment of PWV, which is therefore referred to as Cf-PWV.

[0008] Augmentation index (AIx) assessment is another useful tool for the measurement of arterial stiffness that has been developed over the past decades. Using the same applanation tonometry technique as PWV, AIx is a non-invasive method that estimates arterial pulse wave reflection and expresses it as a percentage. PWV and AIx are two different indices of arterial stiffness and are not interchangeable.

[0009] The putative mechanism of AIx and its relationship to CVD and atherosclerosis has not yet been fully elucidated. AIx has been shown to increase significantly with age, following a curvilinear pattern. However, it has been suggested that the increase in AIx may be related to the increase in ROS levels, which constitutes one of the early stages in the development of atherosclerosis. Indeed, one study showed that AIx is associated with ROS concentrations in a population of smokers. The study also observed that older age and hypertension are associated with an increase in AIx.

[0010] (Poly)phenols (PPs) are known to have antioxidant effects and may therefore be beneficial against ROS. PPs are secondary metabolites of plants that contain one or more phenolic rings in their structure and range in mass from 300 to 3000 Da, and even up to 20000 Da for larger compounds. More than 8000 different types of PPs have been identified in plants.

[0011] They are highly abundant in fruits and vegetables, as well as in plant-based foods and beverages such as coffee, tea, nuts, olive oil, soy products or cocoa. In recent years, these compounds have attracted much attention and been widely studied due to their potential health benefits. Epidemiological and clinical studies have shown that PPs have the potential to regulate physiopathological conditions and thereby reduce the risk of chronic diseases such as CVD and dementia.

[0012] PPs are one of the most abundant and major groups of phytochemicals, which also include terpenoids (such as carotenoids), alkaloids and sulfur compounds. They are synthesized by plants to protect them against UV-mediated oxidative stress, to strengthen cell walls, to repel herbivores and infections, and to attract pollinators.

[0013] PPs are found in nature in their glycosidic form and less frequently as aglycones or genins (i.e., without the glycosyl moiety). They are classified into different groups based on the number of phenyl rings and their structure. Thus, PPs can be divided into two main groups: flavonoids and nonflavonoids.

[0014] Chemically, flavonoids relate to a 15-carbon skeleton that consists of two phenyl rings, A and B, and one oxygen-containing heterocyclic ring designated C. This structure is also known as "C6-C3-C6". Usually, the B ring is attached to the 2-position of the C ring, but it can also be found in the 3-position, as in the case of isoflavones. Flavonoids are divided into seven subclasses: flavones, flavanones, anthocyanins, flavonols, flavan-3-ols, isoflavones, and dihydrochalcones.

[0015] Non-flavonoid families include stilbenes, lignans, phenolic acids, and other PPs such as tyrosol, pyrogallol, or hydroxycoumarins (Manach et al. 2004).Phenolic acids are divided into five subgroups: hydroxyphenylpropanoic acids, hydroxyphenylacetic acids, hydroxyphenylpentanoic acids, hydroxycinnamic acids, and hydroxybenzoic acids (Phenol Explorer).

[0016] Most PPs are ingested orally; therefore, the in vivo digestion of any PP will have a direct effect on the activity of the ingested PP. Often considered a "second brain", the microbiome is a large ecosystem made up of trillions of bacteria that interact with the entire organism throughout an individual's lifetime.

[0017] The intestinal microflora plays an important role in the bioavailability of PP, since the presence of polymeric compounds, associated with a relatively low absorption of phenolic compounds, generally favors the interaction of the latter with colonic bacteria.

[0018] Upon reaching the large intestine, a bidirectional relationship will occur between PP and the microbiota: indeed, PP can regulate the composition and diversity of the microbiome, while gut bacteria catabolize PP to produce smaller compounds that are usually more active and exhibit better absorption than the original metabolites.

[0019] Therefore, the microbiome can play an important role in maintaining the physiological functions of the body. Dysbiosis of the microbiome can lead to various disorders. Microbe-based therapies can be used for maintaining gut health and treating microbiome-related disorders. For example, Jing Li et al. reported that symbiotic imbalance of gut microbiota contributes to the development of hypertension (Li, J., Zhao, et al. 2017,'Gut microbiota dysbiosis contributes to the development of hypertension', Microbiome, 5(1), PP.1-19.)

[0020] Aronia melanocarpa (Aronia) is a berry belonging to the Rosaceae family. Also known as "black chokeberry" due to its astringent taste, the berry is originally native to North America. However, today Aronia is found and cultivated in Central and Eastern Europe (USDA), and since its introduction, various cultivars have been created that exhibit larger berries and better resistance compared to the original cultivars, such as Nero, Viking or Aron. Aronia berries are a rich source of PP and are generally considered to be one of the highest sources of PP among berries. The main PPs found in the berry are represented by procyanidins, anthocyanins and phenolic acids.

[0021] The listing or discussion of an apparently prior-published document in this specification should not necessarily be construed as an acknowledgement that the document is part of the state of the art or is common general knowledge. Summary of the Invention

[0022] Disclosure of the Invention It has now been surprisingly and unexpectedly found by the inventors that an aronia berry extract is capable of treating and / or preventing cardiovascular disease (CVD) and / or gut dysbiosis imbalance.

[0023] Thus, the present invention provides a composition for use in treating and / or preventing cardiovascular disease, wherein the composition comprises an extract obtained or obtainable from Aronia melanocarpa.

[0024] The present invention also provides a composition for use in treating and / or preventing gut microbial dysbiosis, wherein the composition comprises an extract obtained or obtainable from Aronia melanocarpa.

[0025] The present invention also provides a method for treating and / or preventing cardiovascular disease, wherein the method comprises administration of a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0026] The present invention also provides a method for treating and / or preventing gut microbial dysbiosis, the method comprising administering a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0027] The present invention may also provide the use of a composition comprising an extract obtained or obtainable from Aronia melanocarpa in the manufacture of a medicament for the treatment and / or prevention of cardiovascular disease and / or gut dysbiosis imbalance.

[0028] As used herein, treating cardiovascular disease may include at least one of the following: (a) treating and / or preventing hypertension; and / or (b) treating or preventing prehypertension, and / or (b) reducing blood pressure; and / or (c) Reducing arterial stiffness.

[0029] For example, the present invention may provide a composition for use in treating and / or preventing hypertension, lowering blood pressure, and / or reducing arterial stiffness, wherein the composition comprises an extract obtained or obtainable from Aronia melanocarpa. The present invention may also provide a method for treating and / or preventing hypertension, lowering blood pressure, and / or reducing arterial stiffness, wherein the method comprises administration of a composition comprising an extract obtained or obtainable from Aronia melanocarpa to a subject in need thereof.

[0030] The present invention may provide use of a composition comprising an extract obtained or obtainable from Aronia melanocarpa in the manufacture of a medicament for the treatment and / or prevention of hypertension, for the treatment and / or prevention of prehypertension, for lowering blood pressure and / or for reducing arterial stiffness.

[0031] Arterial stiffness has been shown to be associated with increased risk of cardiovascular disease events and cardiovascular death. For example, a 10% increase in Aix has been shown to result in a 48% increase in risk of cardiovascular death (London et al 2001 J Spinal Cord Medicine 32: 72-78). Therefore, it is clear that a reduction in arterial stiffness is interpreted as an indication for the treatment or prevention of CVD or CVD events.

[0032] As used herein, treating gut dysbiosis may include at least one of the following: (a) increased microbiome diversity as measured by fecal microbiome gene counts; (b) increasing the levels of beneficial bacteria, including, but not limited to, one or more of Faecalibacterium prausnitzii 2, Lawsonibacter asaccharolyticus, Intestinimonas butyriciproducens, Faecalibacterium, Roseburia intestinalis, and / or Eggerthella lenta; and / or (c) Reducing the levels of pathogenic microorganisms or microorganisms identified as having no or little benefit against one or more conditions.

[0033] For example, the present invention provides for increasing microbiome diversity as measured by fecal microbiome gene counts and / or increasing the diversity of Clostridiales bacterium, Oscillibacter sp., Firmicutes bacterium CAG 103, Lawsonibacter asaccharolyticus, Oscillospirales 5, Clostridium sp. Clostridium sp., Butyricimonas faecihominis, Turicibacter sanguinis, Bacteroides dorei, Oscillospiraceae, Bacteroides xylanisolvens, Ruminococcus sp. / Blautia sp. Blautia sp., Dialister invisus, Flavonifractor sp., Clostridium sp., Faecalibacterium prausnitzii 2, Christensenellales, Blautia A, Intestinimonas In one embodiment, the composition may be provided for use in increasing the levels of beneficial microorganisms, including, but not limited to, one or more of Saccharomyces cerevisiae, ...

[0034] In preferred embodiments, the bacteria that are increased include one or more of Faecalibacterium prausnitzii 2, Lawsonibacter asaccharolyticus, Intestinimonas butyriciproducens, Faecalibacterium, Roseburia intestinalis and / or Eggerthella lenta. In preferred embodiments, the beneficial bacteria whose levels are increased include one or more of Faecalibacterium prausnitzii 2, Lawsonibacter asaccharolyticus, and / or Intestinimonas butyriciproducens.

[0035] The present invention may also provide a method for increasing microbiome diversity as measured by fecal microbiome gene counts and / or increasing levels of beneficial bacteria, including but not limited to one or more of Faecalibacterium prausnitzii 2, Lawsonibacter asaccharolyticus, Intestinimonas butyriciproducens, Faecalibacterium, Roseburia intestinalis, and / or Eggerthella lenta, comprising administering to a subject in need thereof a composition comprising an extract obtained or obtainable from Aronia melanocarpa. In a preferred embodiment, the beneficial bacteria whose levels are increased include one or more of Faecalibacterium prausnitzii 2, Lawsonibacter asaccharolyticus, and / or Intestinimonas butyriciproducens.

[0036] In some embodiments, the diversity of the fecal microbiome is increased by at least 2%, e.g., at least 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or at least 50%, after treatment with a composition of the invention compared to the diversity of the subject's fecal microbiome before administration of the composition.

[0037] Such compositions or methods for use are hereinafter referred to as "compositions for use according to the invention" or "methods according to the invention."

[0038] Hypertension, also known as high or elevated blood pressure, is a condition in which blood vessels have persistently elevated pressure. Blood is transported from the heart to all parts of the body in blood vessels. With each beat, the heart pumps blood into the vessels. Blood pressure is caused by the force of blood pushing against the walls of the blood vessels (arteries) as it is pumped by the heart. The higher the pressure, the harder the heart has to pump.

[0039] Typically, hypertension is defined as having a systolic blood pressure of 130 or greater, or 140 or greater, and / or a diastolic blood pressure of 80 or greater, or 90 or greater. Prehypertension is defined as having a systolic blood pressure of 120-130, or 120-140, and / or a diastolic blood pressure of 80-90.

[0040] In some embodiments, treating hypertension is defined as a reduction in mm Mercure of at least 2 points, at least 3, 4, 5, 6, 7, 8, 9, 10, 11 points, from pre-treatment values ​​after a period of treatment. In some embodiments, treating pre-hypertension is defined as a reduction in mm Mercure of at least 2 points, at least 3, 4, 5, 6, 7, 8, 9, 10, 11 points, from pre-treatment values ​​after a period of treatment.

[0041] Arterial stiffness is typically measured using pulse wave velocity (PWV) and / or augmentation index (Aix). Therefore, in the present invention, reducing arterial stiffness can be measured by determining PWV and / or AIx. A decrease in PWV and / or AIx indicates a decrease in arterial stiffness.

[0042] In some embodiments of the present invention, treatment with the composition of the present invention results in a reduction in arterial stiffness of 2% or more, for example at least 3%, at least 4%, at least 5%, at least 10%, 15%, 20%, 25%, 30% or more, compared to the arterial stiffness of the subject before treatment with the composition.In some embodiments, the reduction in arterial stiffness is 2% or more, for example at least 3%, at least 4%, at least 5%, or at least 10%, 15%, 20%, 25%, 30%, compared to the arterial stiffness of the subject of a control group.In some embodiments, the control group is represented by a group of individuals known to have high arterial stiffness, for example a group of individuals with cardiovascular disease.Those skilled in the art can select the appropriate composition of the control subjects.

[0043] The diversity of microbiome depends on the number of genetically different bacterial species present in the relevant biome.In the present invention, the biome is the gut microbiome, for example, the gut microbiome as detectable in feces.Therefore, increasing the diversity of the microbiome used in the present invention is intended to mean increasing the number of genetically different bacterial species in the gut as measured by fecal microbiome gene count.For example, in some embodiments, increasing the microbial diversity in the gut comprises increasing the number of genetically different bacterial species in the gut by at least 1%, or at least 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, or at least 40%, compared with the microbial diversity in the gut of an individual before treatment with the composition of the present invention.

[0044] In some embodiments, the microbial diversity of the subject's feces is increased by at least 1%, or at least 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, or at least 40% compared to the microbial diversity present in a population of samples obtained from subjects known to have low microbial diversity.

[0045] In some embodiments, increasing microbial diversity in the gut comprises increasing the number of genes in a fecal sample of a subject by at least 1%, or at least 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, or at least 40% compared to the number of genes in a fecal sample taken from the individual prior to treatment with a composition of the invention.

[0046] In some embodiments, increasing microbial diversity in the gut is achieved when the microbial diversity in a fecal sample obtained from a subject is the same as or substantially similar (i.e., within a statistically relevant range) to the average microbial diversity of a population of healthy subjects.

[0047] In some embodiments, increasing microbial diversity in the gut comprises increasing the number of genes in a fecal sample from a subject by at least 1%, or at least 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, or at least 40% compared to the number of genes in a population of fecal samples taken from a population of subjects known to have low microbial diversity.

[0048] In some embodiments, increased microbial diversity in the gut is achieved when the gene count in a fecal sample obtained from a subject is the same as or substantially similar (i.e., within a statistically relevant range) to the average gene count in a population of healthy subjects.

[0049] In some embodiments, the average number of genes in a population of healthy subjects is about or at least 370,000, about or at least 380,000, 400,000 or at least 400,000, e.g., at least 450,000, 500,000, 550,000 or at least 600,000.

[0050] The type of bacteria present in the gut microbiome also plays an important role in gut health. In the present invention, it has been found that increasing the level of beneficial bacteria, including but not limited to one or more of Faecalibacterium prausnitzii 2, Lawsonibacter asaccharolyticus and Intestinimonas butyriciproducens, Faecalibacterium, Roseburia intestinalis and / or Eggerthella lenta, improves gut health and gut dysbiosis.

[0051] Without being bound by any theory, the increase in gene number and the increase in certain beneficial bacteria can positively affect the state of the intestine and can be used as a biomarker of certain intestinal conditions that can predict certain diseases. For example, Faecalibacterium prausnitzii has been found to be a good biomarker for distinguishing Crohn's disease (CD) and colorectal cancer (CRC) from healthy subjects (Lopez-Siles et al, The ISME Journal, (2017), 11, 841-852).

[0052] The inventors have surprisingly and unexpectedly found that increasing levels of Faecalibacterium prausnitzii 8, Acutalibacteraceae 3, Firmicutes bacterium CAG 103 and / or Bifidobacterium adolescentis can be used to predict prehypertensive subjects who may respond to treatment with the compositions of the present invention, as discussed above.

[0053] It has also been surprisingly found by the inventors that treating cardiovascular diseases and / or gut dysbiosis with an extract obtained or obtainable from Aronia melanocarpa as defined above is more beneficial in a subpopulation of mammals (i.e. humans).

[0054] The population of subjects found to be most likely to respond to treatment with the compositions of the present invention were those with lower fecal microbiome gene counts and relatively high abundance of Faecalibacterium prausnitzii 8, Acutalibacteraceae 3, Firmicutes bacterium CAG 103 and / or Bifidobacterium adolescentis.

[0055] Thus, in one aspect, the present invention provides: 1. A method for determining whether a subject is likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, the method comprising: a) determining the number of fecal microbiome genes in a fecal sample obtained from a subject; and / or b) determining the abundance of any one, two, three, or all of the following: i) Faecalibacterium prausnitzii 8 in fecal samples obtained from subjects; ii) Acutalibacteraceae 3 in fecal samples obtained from subjects; iii) the presence of the bacterium Firmicutes CAG 103 in a fecal sample obtained from the subject; and / or iv) Bifidobacterium adolescentis in a fecal sample obtained from the subject.

[0056] It will be apparent that in some cases, it will be necessary for one of skill in the art to compare the gene counts and / or abundances of particular microorganisms to a control sample(s) or to a standardized value so that a person skilled in the art can determine whether a particular subject is likely to be a responder to a treatment.

[0057] Thus, in some embodiments, the method for determining whether a subject is likely to respond to a treatment further comprises: a) comparing the fecal microbiome gene count in a fecal sample obtained from the subject with the fecal microbiome gene count in at least a first control sample or a population of control samples; and / or b) Compare the following: i) the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from the subject and the abundance of Faecalibacterium prausnitzii 8 in at least a first control sample or a population of control samples; ii) the abundance of Acutalibacteraceae 3 in a fecal sample obtained from the subject and the abundance of Acutalibacteraceae 3 in at least a first control sample or a population of control samples; iii) the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject and the abundance of Firmicutes bacterium CAG 103 in at least a first control sample or a population of control samples; and / or iv) the abundance of Bifidobacterium adolescentis in a fecal sample obtained from the subject and the abundance of Bifidobacterium adolescentis in at least a first control sample or a population of control samples.

[0058] Those skilled in the art will be fully capable of selecting an appropriate control sample, and will be familiar with positive and negative control samples. The following discussion of control samples also applies to other embodiments disclosed herein, for example in the context of arterial stiffness.

[0059] In some embodiments, the control sample is a single control sample, for example taken from a single individual. In other preferred embodiments, the control sample is actually taken from a population of control samples. Those skilled in the art will understand that the statistical power of a method increases with the number of independent samples involved. Therefore, those skilled in the art can select and use an appropriate number of control samples. For example, the population of control samples can be composed of at least 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 500, or at least 1000 different control samples.

[0060] Those skilled in the art will also know the appropriate statistical methods that can be employed on a population of control samples to obtain a single value, such as an average value, that can be directly compared with the test value.Thus, in some embodiments, the control sample(s) is actually a single average value of the relevant parameter (e.g., gene count) obtained from the population of samples.Those skilled in the art can then compare the test value (e.g., test gene count) with the average gene count from the relevant control population, thereby determining whether the test sample (e.g., gene count) is within or outside the statistically significant range of the control value.

[0061] One skilled in the art will understand that it is necessary to compare a test value or sample with at least a positive control value or sample, or at least a negative control value or sample, or preferably both a positive control value or sample and a negative control value or sample.

[0062] The inventors have surprisingly found that subjects with low gene counts and / or higher relative abundance of certain microbial species are more likely to respond to treatment with the compositions of the present invention.Therefore, in this scenario, the positive sample or sample group will be a sample or set of samples taken from a subject known to respond to treatment with the compositions of the present invention.For example, it is important that the sample from the positive control population used in generating a positive control sample set or average value is taken from the subject before administration of the composition.

[0063] Similarly, for negative control samples, the negative sample or population of samples will be a sample or set of samples taken from subjects known to be unresponsive to treatment with the compositions of the invention. As noted above, it is important that the negative control sample set or samples from the negative control population to be used in generating, e.g., an average value, etc., are samples taken from the subject prior to administration of the composition.

[0064] Thus, in one embodiment, at least a first control sample or population of control samples is a negative control sample or population of control samples, or, for example, a statistically relevant value, such as a mean value, obtained from a population of control samples.

[0065] In some embodiments, the negative control sample or population of control samples is a fecal sample collected from one or more negative control subjects who have been determined to be unresponsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the samples were collected from the one or more control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0066] In some embodiments, at least a first control sample or population of control samples is a positive control sample or population of control samples, or is a statistically relevant value, e.g., a mean value, obtained from a population of control samples.

[0067] In some embodiments, the positive control sample or population of control samples is a fecal sample taken from one or more positive control subjects shown to be responsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the samples were taken from the one or more control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0068] In some embodiments, the fecal microbiome gene count of a sample obtained from a subject is compared to the following: a) fecal microbiome gene counts in a negative control sample or a population of control samples, or a statistically relevant value, e.g., a mean value, obtained from a population of control samples, e.g., where the negative control sample or population of control samples is a fecal sample taken from one or more negative control subjects that have been shown to be unresponsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, where the sample was taken from the one or more negative control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; and / or b) fecal microbiome gene counts in a positive control sample or a population of control samples, or a statistically relevant value, e.g., an average value, obtained from a population of control samples, e.g., wherein the positive control sample or population of control samples are fecal samples taken from one or more positive control subjects shown to be responsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, and wherein the samples were taken from the one or more positive control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0069] In some embodiments: a) if the fecal microbiome gene count of the sample obtained from the subject is substantially similar to or higher than the fecal microbiome gene count of the negative control sample(s) or a statistically relevant value, e.g., the mean value, obtained from a population of control samples, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; For example, herein, if the fecal microbiome gene count of a sample obtained from the subject is at least 5% greater, e.g., at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, e.g., at least 200% greater, than the fecal microbiome gene count of a negative control sample(s), then the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; and / or b) if the fecal microbiome gene count in the test sample is substantially similar to or lower than the fecal microbiome gene count of the positive control sample(s) or a statistically relevant value, e.g., the mean value, obtained from a population of control samples, the subject is likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; For example, herein, if the fecal microbiome gene count of a sample obtained from a subject is at least 5% lower, such as at least 10% lower, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, such as at least 200% lower, than the fecal microbiome gene count of a positive control sample(s), then the subject is considered to be likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0070] One of skill in the art can determine whether a particular test value is significantly different from a positive or negative control subject, and can determine whether a particular test subject should be classified as being from a responder or a non-responder.

[0071] In some embodiments: a) i) the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from a subject is compared with the abundance of Faecalibacterium prausnitzii 8 in at least a first control sample or a population of control samples, or with a statistically relevant value, e.g., a mean value, obtained from a population of control samples; ii) the abundance of Acutalibacteraceae 3 in a fecal sample obtained from the subject is compared to the abundance of Acutalibacteraceae 3 in at least a first control sample or a population of control samples, or a statistically relevant value, e.g., a mean value, obtained from a population of control samples; iii) the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from a subject is compared to the abundance of Firmicutes bacterium CAG 103 in at least a first control sample or a population of control samples, or a statistically relevant value, e.g., a mean value, obtained from a population of control samples; and / or iv) the abundance of Bifidobacterium adolescentis in the fecal sample obtained from the subject is compared to the abundance of Bifidobacterium adolescentis in at least a first control sample or a population of control samples, or to a statistically relevant value, e.g., a mean value, obtained from a population of control samples; wherein the control sample or population of control samples is a negative control sample or population of control samples, or a statistically relevant value, e.g., a mean value, obtained from a population of control samples; wherein the negative control sample or population of control samples is a fecal sample taken from one or more negative control subjects that have been shown to be unresponsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the sample is taken from one or more positive control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; and / or b) i) the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from a subject is compared with the abundance of Faecalibacterium prausnitzii 8 in at least a first control sample or a population of control samples, or with a statistically relevant value, e.g., a mean value, obtained from a population of control samples; ii) Acutalibacteraceae 3 in a fecal sample obtained from the subject is compared with the abundance of Acutalibacteraceae 3 in at least a first control sample or a population of control samples, or with a statistically relevant value, e.g., a mean value, obtained from a population of control samples; iii) the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from a subject is compared to the abundance of Firmicutes bacterium CAG 103 in at least a first control sample or a population of control samples, or a statistically relevant value, e.g., a mean value, obtained from a population of control samples; and / or iv) the abundance of Bifidobacterium adolescentis in the fecal sample obtained from the subject is compared to the abundance of Bifidobacterium adolescentis in at least a first control sample or a population of control samples, or to a statistically relevant value, e.g., a mean value, obtained from a population of control samples; wherein the control sample or population of control samples is a positive control sample or population or control sample; Here, the positive control sample or population of control samples is a fecal sample taken from one or more positive control subjects shown to be responsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the samples were taken from the one or more positive control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0072] In some embodiments: a) a subject is considered likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa if the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from the subject is substantially similar to or higher than the abundance of Faecalibacterium prausnitzii 8 in a positive control sample(s) or a statistically relevant value, e.g., a mean value, obtained from a population of control samples, optionally wherein the abundance of Faecalibacterium prausnitzii 8 in a sample obtained from the subject is substantially similar to or higher than the abundance of Faecalibacterium prausnitzii 8 in a positive control sample(s) or a statistically relevant value, e.g., a mean value, obtained from a population of control samples; 8, or a statistically relevant value, e.g., the mean value, obtained from a population of control samples is at least 5% greater, e.g., at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, e.g., at least 200% greater, the subject is considered likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; b) the subject is deemed likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa if the abundance of Acutalibacteraceae 3 in a fecal sample obtained from the subject is substantially similar to or higher than the abundance of Acutalibacteraceae 3 in the positive control sample(s) or a statistically relevant value, e.g., the mean value, obtained from a population of control samples, optionally wherein the abundance of Acutalibacteraceae 3 in the sample obtained from the subject is substantially similar to or higher than the abundance of Acutalibacteraceae 3 in the positive control sample(s) or a statistically relevant value, e.g., the mean value, obtained from a population of control samples; 3, or a statistically relevant value, e.g., the mean value, obtained from a population of control samples is at least at least 5% greater, e.g., at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, e.g., at least 200% greater, the subject is considered likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; c) the subject is likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa if the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject is substantially similar to or higher than the abundance of Firmicutes bacterium CAG 103 in a positive control sample(s) or a statistically relevant value, e.g., the mean value, obtained from a population of control samples, optionally wherein the abundance of Firmicutes bacterium CAG 103 in a sample obtained from the subject is substantially similar to or higher than the abundance of Firmicutes bacterium CAG 103 in a positive control sample(s) or a statistically relevant value, e.g., the mean value, obtained from a population of control samples; a subject is considered likely to be responsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa if the abundance of 103, or a statistically relevant value, e.g., the mean, obtained from a population of control samples is at least 5% greater, e.g., at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, e.g., at least 200% greater; and / or d) the subject is likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa if the abundance of Bifidobacterium adolescentis in a fecal sample obtained from the subject is substantially similar to or higher than the abundance of Bifidobacterium adolescentis in a positive control sample(s) or a statistically relevant value, e.g., a mean value, obtained from a population of control samples, optionally wherein the abundance of Bifidobacterium adolescentis in the sample obtained from the subject is substantially similar to or higher than the abundance of Bifidobacterium adolescentis in the positive control sample(s) or a statistically relevant value, e.g., a mean value, obtained from a population of control samples. adolescentis abundance, or a statistically relevant value, e.g., the mean, obtained from a population of control samples is at least 5% greater, e.g., at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, e.g., at least 200% greater, the subject is considered likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; e) if the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from the subject is substantially similar to or lower than the abundance of Faecalibacterium prausnitzii 8 in the negative control(s) or a statistically relevant value, e.g., a mean value, obtained from a population of control samples, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Faecalibacterium prausnitzii 8 in a sample obtained from the subject is substantially similar to or lower than the abundance of Faecalibacterium prausnitzii 8 in the negative control sample(s) or a statistically relevant value, e.g., a mean value, obtained from a population of control samples; a subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa if the abundance of 8 or a statistically relevant value, e.g., the mean, is at least 5% lower, e.g., at least 10% higher, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, e.g., at least 200% lower than the mean value of 8 or a statistically relevant value, e.g., the mean value, obtained from a population of control samples; f) if the abundance of Acutalibacteraceae 3 in a fecal sample obtained from the subject is substantially similar to or lower than the abundance of Acutalibacteraceae 3 in the negative control sample(s) or a statistically relevant value, e.g., the mean value, obtained from a population of control samples, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Acutalibacteraceae 3 in a sample obtained from the subject is substantially similar to or lower than the abundance of Acutalibacteraceae 3 in the negative control sample(s) or a statistically relevant value, e.g., the mean value, obtained from a population of control samples; 3, or a statistically relevant value, e.g., the mean value, obtained from a population of control samples is at least at least 5% lower, e.g., at least 10% lower, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, e.g., at least 200% lower, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; g) if the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject is substantially similar to or lower than the abundance of Firmicutes bacterium CAG 103 in a negative control sample(s) or a statistically relevant value, e.g., a mean value, obtained from a population of control samples, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Firmicutes bacterium CAG 103 in a sample obtained from the subject is substantially similar to or lower than the abundance of Firmicutes bacterium CAG 103 in a positive control sample(s), or a statistically relevant value, e.g., a mean value, obtained from a population of control samples, an abundance of 103, or a statistically relevant value, e.g., the mean, obtained from a population of control samples is at least 5% lower, e.g., at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, e.g., at least 200% lower, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; and / or h) if the abundance of Bifidobacterium adolescentis in a fecal sample obtained from the subject is substantially similar to or lower than the abundance of Bifidobacterium adolescentis in a negative control sample(s) or a statistically relevant value, e.g., the mean, obtained from a population of control samples, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Bifidobacterium adolescentis in a sample obtained from the subject is substantially similar to or lower than the abundance of Bifidobacterium adolescentis in a positive control sample(s) or a statistically relevant value, e.g., the mean, obtained from a population of control samples. adolescentis abundance, or a statistically relevant value, e.g., the mean, obtained from a population of control samples, is at least 5% lower, e.g., at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, e.g., at least 200% lower, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0073] In some preferred embodiments, the abundance of each of the above microorganisms is determined. Thus, in some embodiments: i) the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from a subject is compared with the abundance of Faecalibacterium prausnitzii 8 in at least a first control sample or a population of control samples, or with a statistically relevant value, e.g., a mean value, obtained from a population of control samples; ii) the abundance of Acutalibacteraceae 3 in a fecal sample obtained from the subject is compared to the abundance of Acutalibacteraceae 3 in at least a first control sample or a population of control samples, or a statistically relevant value, e.g., a mean value, obtained from a population of control samples; iii) the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from a subject is compared to the abundance of Firmicutes bacterium CAG 103 in at least a first control sample or a population of control samples, or a statistically relevant value, e.g., a mean value, obtained from a population of control samples; and iv) The abundance of Bifidobacterium adolescentis in a fecal sample obtained from a subject is compared with the abundance of Bifidobacterium adolescentis in at least a first control sample or a population of control samples, or a statistically relevant value, e.g., the mean value, obtained from a population of control samples.

[0074] The abundance of Bifidobacterium adolescentis is believed to be particularly useful in indicating whether a subject will respond to treatment. Thus, in one embodiment, the method comprises determining the abundance of Bifidobacterium adolescentis in a fecal sample obtained from a subject, and determining whether the abundance of Bifidobacterium adolescentis is in a fecal sample obtained from a subject, and determining whether the abundance of Bifidobacterium adolescentis is in a fecal sample obtained from a subject, as follows: the abundance of Bifidobacterium adolescentis in one or more negative control samples, or a statistically relevant value, e.g., the mean value, obtained from a population of control samples; and / or The abundance of Bifidobacterium adolescentis in one or more positive control samples, or a statistically relevant value obtained from a population of control samples, e.g., the mean This includes comparing with.

[0075] In some embodiments, the method for determining whether a subject is likely to respond to treatment with the composition of the present invention simply involves determining the fecal microbiome gene count in a fecal sample obtained from the subject. In some embodiments, if the fecal microbiome gene count is less than 400,000, optionally less than 375,000, optionally less than 350,000, 300,000, 275,000, 250,000, 225,000, 200,000, less than 175,000, or less than 150,000, the subject is considered likely to respond to treatment with the composition of the present invention.

[0076] As described elsewhere herein, one method for determining the number of microbiome genes is by shotgun sequencing. Thus, in some embodiments of any aspect or embodiment of the present invention, the number of fecal microbiome genes in a fecal sample is determined by shotgun sequencing.

[0077] Similarly, in some embodiments of any aspect or embodiment of the invention, i) Faecalibacterium prausnitzii 8 in fecal samples obtained from subjects; ii) Acutalibacteraceae 3 in fecal samples obtained from subjects; iii) Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject; and / or iv) Bifidobacterium adolescentis in fecal samples obtained from subjects To determine the abundance of is performed using shotgun sequencing.

[0078] In some embodiments, responding to treatment with a composition of the invention is intended to mean responding by exhibiting a reduction in blood pressure. Thus, in one embodiment, the response to treatment is exhibited by a reduction in blood pressure.

[0079] For example, the present invention provides methods for determining whether a composition comprising an extract obtained or obtainable from Aronia melanocarpa is likely to reduce blood pressure in a particular subject, including methods for determining whether a subject is likely to respond to treatment with a composition of the present invention as described herein.

[0080] In some embodiments of the methods for determining whether a composition comprising an extract obtained or obtainable from Aronia melanocarpa is likely to reduce blood pressure in a particular subject and / or whether the subject is likely to respond to treatment with the composition, the subject is prehypertensive, e.g., the subject is: a systolic blood pressure of at least 120 mmHg, optionally at least 130 or at least 140 mmHg; and / or a systolic blood pressure of between 120 mmHg and 130 mmHg, or between 120 mmHg and 140 mmHg, and / or A diastolic blood pressure of at least 80mmHg, optionally at least 90mmHg; and / or a diastolic blood pressure between 80mmHg and 90mmHg. has.

[0081] The present invention therefore also provides a composition for use in treating and / or preventing cardiovascular disease and / or gut dysbiosis imbalance in a subject, wherein the composition comprises an extract obtained or obtainable from Aronia melanocarpa, and wherein the subject has been determined to be likely to respond to treatment with a composition according to any of the methods of the present invention.

[0082] The present invention also provides a method for treating and / or preventing cardiovascular disease and / or gut dysbiosis imbalance in a subject, the method comprising administering to a subject in need thereof a composition comprising an extract obtained or obtainable from Aronia melanocarpa, and wherein the subject has been determined to be likely to respond to treatment with a composition according to any of the methods of the present invention.

[0083] Accordingly, the present invention also provides a composition for use in treating hypertension, prehypertension, and / or alleviating hypertension in a subject, wherein the composition comprises an extract obtained or obtainable from Aronia melanocarpa, and the subject has been determined to be likely to respond to treatment with the composition according to any of the methods of the present invention.

[0084] The present invention also provides a method for reducing hypertension, prehypertension and / or hypertension in a subject, the method comprising administering to a subject in need thereof a composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the subject has been determined to be likely to respond to treatment with the composition according to any of the methods of the present invention.

[0085] The present invention may also provide a composition for use in treating gut dysbiosis imbalance in a mammal (i.e., a human) having a fecal microbiome gene count of less than 400,000, optionally less than 375,000, optionally less than 350,000, 300,000, 275,000, 250,000, 225,000, 200,000, 175,000, or less than 150,000, wherein the composition comprises an extract obtained or obtainable from Aronia melanocarpa.

[0086] The present invention may also provide a method for treating gut dysbiosis imbalance in a mammal (i.e., a human) having a fecal microbiome gene count of less than 400,000, optionally less than 375,000, optionally less than 350,000, 300,000, 275,000, 250,000, 225,000, 200,000, 175,000, or less than 150,000, wherein the method comprises administration of a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0087] The invention may also provide for the use of a composition comprising an extract obtained or obtainable from Aronia melanocarpa in a method of manufacture of a medicament for use in treating gut dysbiosis imbalance in a mammal (i.e., human) having a fecal microbiome gene count similar to or lower than the fecal microbiome gene count of a positive control sample(s), or a statistically relevant value, e.g., mean value, obtained from a population of control samples, e.g., wherein the fecal microbiome gene count of a sample obtained from the subject is at least 5% lower, e.g. at least 10% lower, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, e.g. at least 200% lower, than the fecal microbiome gene count of the positive control sample(s).

[0088] The present invention may also provide for the use of a composition comprising an extract obtained or obtainable from Aronia melanocarpa in a method of manufacture of a medicament for use in treating a gut dysbiosis imbalance in a mammal (i.e., a human) having a fecal microbiome gene count of less than 400,000, optionally less than 375,000, optionally less than this.

[0089] The present inventors have surprisingly found that supplementation with Aronia melanocarpa results in an increase in microbiome diversity in subjects, more specifically an increase in the bacteria shown in FIG.

[0090] The present invention also relates to increasing microbiome diversity as measured by fecal microbiome gene counts in subjects having fecal microbiome gene counts similar to or lower than the fecal microbiome gene counts in a positive control(s) or a statistically relevant value, e.g., the mean, obtained from a population of control samples, and / or increasing the ... in subjects having fecal microbiome gene counts in subjects having fecal microbiome gene counts in subjects having fecal micro 103 / A composition may be provided for use in increasing the level of one or more of Clostridium sp., Dysomobacter welbionis, Ruminococcus sp., Clostridiales bacterium, bacteria of the Clostridiaceae family, for example wherein the fecal microbiome gene count of a sample obtained from a mammal is at least 5% lower, such as at least 10% lower, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, such as at least 200% lower, than the fecal microbiome gene count of a positive control sample(s), wherein the composition comprises an extract obtained or obtainable from Aronia melanocarpa.

[0091] In a preferred embodiment, the increased bacteria are selected from the beneficial bacteria Faecalibacterium, Roseburia intestinalis and / or Eggerthella lenta.

[0092] The present invention also relates to increasing microbiome diversity as measured by fecal microbiome gene count in subjects having a fecal microbiome gene count of less than 400,000, optionally less than 375,000, optionally less than 350,000, 300,000, 275,000, 250,000, 225,000, 200,000, 175,000, or less than 150,000, and / or increasing the ... lactatiformans, Lachnoclostridium spp. / Clostridium sp., Acutalibacteraceae 3, Clostridium sp., Collinsella bouchesdurhonensis, Firmicutes bacterium CAG 103 / Clostridium sp., Dysomobacter welbionis, Ruminococcus sp., Clostridiales bacterium, Clostridiales bacteria, wherein the composition comprises an extract obtained or obtainable from Aronia melanocarpa. In a preferred embodiment, the increased bacteria is selected from the beneficial bacteria Faecalibacterium, Roseburia intestinalis and / or Eggerthella lenta.

[0093] The present invention may also provide a composition for use in increasing microbiome diversity as measured by fecal microbiome gene count and / or increasing levels of beneficial bacteria such as Faecalibacterium, Roseburia intestinalis and / or Eggerthella lenta in a mammal (i.e., human) having a fecal microbiome gene count of less than 400,000, optionally less than 375,000, optionally less than 350,000, 300,000, 275,000, 250,000, 225,000, 200,000, 175,000, or less than 150,000, wherein the composition comprises an extract obtained or obtainable from Aronia melanocarpa.

[0094] The present invention may also provide a method for increasing microbiome diversity as measured by fecal microbiome gene count and / or increasing levels of beneficial bacteria such as Faecalibacterium, Roseburia intestinalis and / or Eggerthella lenta in a mammal (i.e., human) having a fecal microbiome gene count of less than 400,000, optionally less than 375,000, optionally less than 350,000, 300,000, 275,000, 250,000, 225,000, 200,000, 175,000, or less than 150,000, wherein the method comprises administration of a composition comprising an extract obtained or obtainable from Aronia melanocarpa to a subject in need thereof.

[0095] The present invention also provides methods for increasing microbiome diversity as measured by fecal microbiome gene counts in mammals (i.e., humans) having fecal microbiome gene counts similar to or lower than the fecal microbiome gene counts in a positive control(s) or a statistically relevant value, e.g., the mean, obtained from a population of control samples, and / or for detecting Faecalibacterium, Roseburia intestinalis, and / or Eggerthella A method for increasing levels of beneficial bacteria such as Aronia melanocarpa may be provided, for example wherein the fecal microbiome gene count of a sample obtained from a mammal is at least 5% lower, e.g., at least 10% lower, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, e.g., at least 200% lower, than the fecal microbiome gene count of a positive control sample(s), wherein the method comprises administration of a composition comprising an extract obtained or obtainable from Aronia melanocarpa.

[0096] As will be understood by one of skill in the art, the term "obtainable from" as used herein means that the extract may be obtained or isolated from the plant, or may be obtained from an alternative source, for example, by chemical synthesis or enzymatic production, whereas the term "obtained" as used herein means that the extract is obtained directly from the plant source.

[0097] The extract obtained or obtainable from Aronia melanocarpa may be in liquid or solid form. Typically, the extract is in solid, i.e., powder form.

[0098] All references herein to extracts obtained or obtainable from Aronia melanocarpa typically refer to extracts obtained or obtainable from the fruit juice of Aronia melanocarpa, which may be concentrated.

[0099] In some aspects, the juice of Aronia melanocarpa (juice concentrate) may be extracted using only water, and this extract may be referred to as a water extract. In another aspect, the juice (or juice concentrate) of Aronia melanocarpa may be extracted with an alcohol, such as ethanol, and this extract may be referred to as an alcohol extract, such as an ethanol extract.

[0100] In a preferred aspect, the Aronia melanocarpa juice (concentrated juice) may be extracted with a mixture of alcohol and water, such as ethanol and water, which may be referred to as a hydroalcoholic extract, such as a hydroethanolic extract.

[0101] In another aspect, the Aronia melanocarpa juice (concentrated juice) may be extracted with a non-alcoholic organic solvent, such as acetone, and this extract may be referred to as an organic extract or an acetone extract.

[0102] Additionally, the polyphenols of the Aronia melanocarpa concentrated juice (or any of the aforementioned extracts) may be further purified to obtain a polyphenol-enriched extract, for example, using a polyphenol absorption column, or any other technique known in the art that provides polyphenols of high purity.

[0103] Typically, the target polyphenols are absorbed by the resin, which allows the remaining solids to pass through the column. Water and ethanol are then used to obtain an eluate, which is then concentrated to obtain the native extract.

[0104] Typically, the extract obtained or derived from Aronia melanocarpa comprises 10% or more total polyphenols (based on catechins) by weight of the extract, such as 20% or more, such as 30% or more, such as 40% or more. In a preferred embodiment, the total polyphenols (based on catechins) is 40% or more.

[0105] The composition containing the extract obtained or obtainable from Aronia melanocarpa may consist of the extract or consist essentially of the extract, or may contain the extract in combination with non-toxic pharma- ceutically acceptable excipients (or ingredients). These excipients (or ingredients) may be, for example: inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as corn starch, maltodextrin or alginic acid; binding agents, such as starch, gelatin or gum arabic; or lubricating agents, such as magnesium stearate, stearic acid, talc and mixtures thereof. In a preferred embodiment, the product does not have a carrier.

[0106] For the avoidance of doubt, preferences, options, specific features and the like set forth with respect to a given aspect of the invention are to be considered as being disclosed in combination with any or all other preferences, options, specific features and the like as set forth with respect to the same or other aspects, features and parameters of the invention, unless the context indicates otherwise.

[0107] When we use the term "consisting essentially of" or "consists essentially of", we mean that the composition or extract or juice being described must include the listed component(s) and may also include small amounts (e.g., up to 2% by weight, or up to 1% by weight, or up to 0.1% by weight, or up to 0.01% by weight) of other components, provided that any additional components do not affect the essential properties of the composition or extract. When we use the term "consisting of", we mean that the composition being described must include only the listed component(s). These terms can be applied in an analogous manner to processes, methods and uses.

[0108] The extract obtained or obtainable from Aronia melanocarpa contains / comprises polyphenols.

[0109] Typically, in a composition comprising an extract obtained or obtainable from Aronia melanocarpa, the total polyphenols (based on catechins) is 10% or more by weight of the composition, such as 20% or more, 30% or more, such as 40% or more by weight. In a preferred embodiment, the total polyphenols (based on catechins) is 40% or more.

[0110] In any of the compositions and methods for use of the present invention, the dosage of the composition for treating cardiovascular disease and / or gut dysbiosis imbalance may be about 100 to about 1000 mg / day by weight of the composition, for example about 400 to about 600 mg / day or about 500 mg / day by weight of the composition.

[0111] In some embodiments, the dosage for treating cardiovascular disease and / or gut dysbiosis imbalance is as follows: a) from about 100 to about 1000 mg / day, optionally from 100 to 1000 mg / day, or from 200 to 800 mg / day, or from 400 to 600 mg / day, by weight of the composition; b) 1000 mg / day or less, optionally 900 mg / day or less, 800 mg / day, 700 mg / day, 600 mg / day, 500 mg / day, 400 mg / day, 300 mg / day, 200 mg / day or less, or 100 mg / day or less, by weight of the composition; and / or c) 100 mg / day or more, or optionally 200 mg / day or more, 300 mg / day, 400 mg / day, 500 mg / day, 600 mg / day, 700 mg / day, 800 mg / day or more, 900 mg / day or more, or 1000 mg / day or more.

[0112] Dosage of the composition may typically provide from about 100 mg / day to about 500 mg / day of polyphenols, for example about 200 mg / day, by weight of the composition.

[0113] In some embodiments, the dosage for treating cardiovascular disease and / or gut dysbiosis provides: a) from about 100 mg / day to about 500 mg / day of polyphenol by weight of the composition, optionally from about 200 mg / day to about 400 mg / day, optionally about 300 mg / day of polyphenol by weight of the composition; b) no more than 500 mg / day of polyphenols by weight of the composition, optionally no more than 400 mg / day, 300 mg / day, 200 mg / day, or no more than 100 mg / day of polyphenols by weight of the composition; and / or c) 100 mg / day or more, optionally 200 mg / day or more, 300 mg / day, 400 mg / day, or 500 mg / day or more of polyphenols by weight of the composition.

[0114] The compositions may be taken once a day or more than once a day depending on the dosage required. Typically, the composition may be taken for at least 1 week, such as at least 6 weeks or at least 12 weeks.

[0115] In the above defined method, determining the fecal microbiome gene count may comprise performing a shotgun test on the fecal sample. Typically, in the method as defined above, the method comprises identifying the subject as likely to respond to treatment if the gene number is less than or equal to 400,000.

[0116] In the compositions and methods for use of the present invention, treating cardiovascular disease and / or gut dysbiosis imbalance as defined above may preferably be in a mammal (i.e., a human). As used in this specification and claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, "a microbe" can include a plurality of microbes.

[0117] The terms "microbiome," "microbiota," and "microbial habitat" may be used interchangeably herein and may refer to the ecological community of microorganisms that live on or within a subject. In the present invention, the microbiome may particularly relate to those found in the gastrointestinal tract.

[0118] The terms "treatment" and "treating" as used herein refer to an approach to obtain beneficial or desired results, including but not limited to therapeutic benefit and / or preventive benefit.Therapeutic benefit refers to a therapeutically appropriate improvement or effect on one or more diseases, conditions or symptoms being treated.For preventive benefit, the composition can be administered to a subject at risk of developing a particular disease, condition or symptom, or to a subject reporting one or more physiological symptoms of a disease, even though the disease, condition or symptom may not yet be manifested.

[0119] As used herein, "administer", "administering", "administration" and their derivatives refer to methods that may be used to allow delivery of an agent or composition to the site where a biological effect is desired. These methods include, but are not limited to, parenteral administration (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular, intrathecal, intranasal, intravitreal, infusion and local injection), transmucosal injection, oral administration, administration as a suppository, and topical administration. In the present invention, the preferred route of administration may be oral administration.

[0120] The term "effective amount" or "therapeutically effective amount" refers to an amount of a composition, such as a composition comprising a microorganism of the present disclosure, that is sufficient to provide a desired activity upon administration to a subject in need thereof. Within the context of the present disclosure, the term "therapeutically effective" refers to an amount of a composition that is sufficient to delay the onset of, halt the progression of, or relieve or alleviate at least one symptom of the disorder treated by the method of the present disclosure. figure [Brief description of the drawings]

[0121] [Figure 1] Study design of the clinical trial. ABPM: ambulatory blood pressure measurement; FMD: flow-mediated dilation; IPAQ: International Standardized Physical Activity Questionnaire.

[0122] [Diagram 2] Box plots showing changes from baseline in gut microbiome gene counts after 12 weeks of intervention. Significance was tested by applying unpaired Wilcoxon rank-sum tests for changes at week 12 compared to baseline in the Aronia vs. control groups. P values ​​represent change from control.

[0123] [Diagram 3]Bar graph of species significantly different between the Aronia and control groups after change from baseline. The bar graph shows the effect size of Cliff delta (absolute value >0.2), with higher color intensity representing larger Cliff delta values.

[0124] [Figure 4] Box plots showing statistical differences between the Aronia responder (R), non-responder (NR), and control (C) groups in changes in 24-h and awake SBPbr and 24-h and awake DBPbr after 12 weeks of supplementation. For each comparison, significance was tested by applying the Kruskal Wallis test with Bonferroni adjustment. SBPbr: brachial systolic blood pressure, DBPbr: brachial diastolic blood pressure.

[0125] [Diagram 5] Box plots showing statistical differences in gut microbiome gene counts at baseline among Aronia responders (R), non-responders (NR), and controls (C). Significance was tested by applying the Kruskal Wallis test and Bonferroni adjustment followed by Dunn post-hoc test.

[0126] [Figure 6-1] Wedge plots showing bacterial taxa significantly enriched in Aronia responders (R), non-responders (NR) and controls (C) at baseline. In each comparison, an upturned triangle indicates that the species is enriched in the first mentioned group (NR for a and b, R for c). Signed effect sizes are shown throughout the marker orientation, and color, hue and size represent absolute effect sizes. Solid borders indicate significance. Markers not shown had no differences in the statistical analysis. [Figure 6-2]Wedge plots showing bacterial taxa significantly enriched in Aronia responders (R), non-responders (NR) and controls (C) at baseline. In each comparison, an upturned triangle indicates that the species is enriched in the first mentioned group (NR for a and b, R for c). Signed effect sizes are shown throughout the marker orientation, and color, hue and size represent absolute effect sizes. Solid borders indicate significance. Markers not shown had no differences in the statistical analysis.

[0127] [Figure 7] Spearman correlation plot between baseline levels of B. adolescentis and chronic changes in 24-h DBP (Aronia: triangles, control: circles). On the right are box plots reporting delta changes in 24-h DBP. On the top are box plots for the log10 of baseline levels of B. adolescentis.

[0128] [Figure 8] Box plots showing the change from baseline in gut microbiome gene counts after 12 weeks of intervention in the Aronia responder (R), non-responder (NR), and control (C) groups. The significance of each comparison was tested by applying the Kruskal Wallis test and Dunn post-hoc test after Bonferroni adjustment.

[0129] [Figure 9] Species significantly different between R and NR after 12 weeks of intervention. Bar graphs show Cliff delta values ​​(an indicator of effect size) of all species that were significantly and relevantly contrasted (absolute value of Cliff delta higher than 0.3). Higher color intensity represents larger absolute value of Cliff delta. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0130] The invention will now be further described with reference to the following non-limiting examples: example

[0131] Example 1 - Preparation of a composition comprising an extract obtained or obtainable from Aronia melanocarpa. Frozen Aronia melanocarpa juice concentrate is diluted with water and applied to a resin column that absorbs polyphenols. The target polyphenols are absorbed by the resin allowing the remaining solids to pass through the column. Water and ethanol are then used to obtain an eluate, which is then concentrated to obtain the native extract. The polyphenol concentration of the extract is approximately 40%.

[0132] The native extract is then spray dried and combined with maltodextrin. The dried extract is then sieved and packaged.

[0133] Example 2 - Investigating the effects of Aronia melanocarpa extract on cardiometabolic health A two-group, double-blind, parallel, randomized, controlled trial was conducted. Participants with a systolic blood pressure of 120-140 mmHg and / or a diastolic blood pressure of 80-90 mmHg participated in four study visits defined as follows: pre-visit 1, visit 1, pre-visit 2, visit 2. A summary of the study design can be found in Figure 1.

[0134] Research method The pre-visit took place 24 hours before visits 1 and 2. During the pre-visit, participants were given a bottle to collect their urine over a 24-hour period and had a 24-hour ambulatory BP monitor (ABPM) attached to their non-dominant arm, during which the first measurement was taken.

[0135] Patients were also asked to complete a 7-day food diary prior to the visit, to avoid caffeine, alcohol, strenuous exercise and tobacco one hour before each visit, and to fast for 12 hours before visits 1 and 2.

[0136] Measurements of peripheral laboratory BP, FMD, PWV, AIx, as well as blood samples were all taken at baseline (0 hours) and then again 2 hours after acute ingestion of the interventional product, at visits 1 and 2. Fecal samples were collected during both visits 1 and 2 and immediately stored at -80°C.

[0137] Participants were instructed to take one capsule of a composition (500 mg) containing an extract derived or obtainable from Aronia melanocarpa or a placebo every morning with a glass of water, ideally with a meal. The first capsule was delivered after the first measurement at Visit 1, which represented the start of the 12-week intake. The last capsule was also taken on the final day (Visit 2) within the study unit, after the first set of deficit measurements.

[0138] Subjects were followed up via email monthly throughout the 12 weeks to ensure good compliance and to record any potential adverse events. The primary outcomes of this study were the effects of aronia berry extract (Aronia) versus placebo (control) on 24-h SBP and DBP during free exercise after 12 weeks of supplementation.

[0139] Secondary endpoints included the effect of the extract compared to placebo on clinic BP, clinic and 24-hour heart rate, arterial stiffness (measured as PWV and AIx), blood lipids (total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides), blood cortisol levels after 12 weeks of daily intake, as well as the safety and tolerability of the aronia berry extract.

[0140] Secondary objectives also included investigating the effects of Aronia extract versus placebo on FMD and blood flow velocity at 2 hours and 12 weeks after ingestion.

[0141] Tertiary outcomes included analysis of blood samples collected at all time points to assess Aronia PP metabolites, analysis of 24-h urine samples to investigate excretion of PP metabolites, and analysis of the gut microbiome of fecal samples after 12 weeks of Aronia berry extract or placebo intake.

[0142] The trial was conducted in accordance with the guidelines laid down in the current version of the Declaration of Helsinki. All procedures were approved by the Ethics Committee of King's College London (RESCM-17 / 18-5283) and the trial was registered at ClinicalTrials.gov under the reference number NCT03434574.

[0143] Twenty-four-hour free exercise peripheral (brachial) and central (aortic) BP, and heart rate (HR) were measured using an Arteriograph24™ (TensioMed, Budapest, Hungary) (Figure 2.1). Twenty-four-hour SBP, DBP, and HR were measured every 30 minutes over a 24-hour period, separating cuff application and data collection.

[0144] Office peripheral (upper arm) BP and heart rate in the supine position were measured after 10 min on the right upper arm in the supine position, in a quiet room, with the arm at heart level, legs uncrossed and supported, back supported, and bladder empty, according to the recommendations of the American Heart Association (Muntner et al. 2019), using an automated clinical digital sphygmomanometer OMRON M3 (OMRON Healthcare UK Ltd, Milton Keynes, UK).

[0145] 24-h PWV and AIx were measured every 30 minutes over 24 hours, pre-visit and during the separate visits, using the Arteriograph24™. Participants were asked to complete a 24-h activity log, indicating their daily activity levels and assessing periods of wakefulness and sleep.

[0146] Volunteers were instructed to lie down and remain silent during the measurements. PWV (units: m / s) was assessed via applanation tonometry using SphygmoCor® (Smart Medical, Gloucestershire, UK). Briefly, PWV is derived from the chain of pulse transit times from the heart to the femoral artery and from the heart to the carotid artery. The device's sensors capture the time lapse between pulse waves in both arteries, allowing the wave velocity to be determined by measuring the distance from the carotid artery to the heart and the distance from the heart to the femoral artery with a tape measure beforehand. Results with a standard deviation of less than 8% were included in the analysis.

[0147] The ascending pressure waveform originates at the aortic level, with a reflected wave that depends on the vascular structure and the general peripheral resistance. As this latter falls at the body periphery, the pressure decreases from the aorta to the wrist. Thus, the waveform has a different shape when measured at the aortic level compared to the wrist. AIx is measured as the ratio between the central pulse pressure at the aortic level and the reflected pulse pressure (called augmentation pressure). AIx is a relevant indicator of the state of arterial stiffness of an individual's vasculature.

[0148] Fecal samples were collected in OMNIgene GUT self-collection tubes (DNA Genotek Inc., Ottawa, Canada) as close as possible to each study visit and stored at -80°C until further analysis. Participants were asked to record the date of collection.

[0149] Microbiome shotgun metagenomics analysis was performed by CosmosID, Inc. (Rockville, MD, USA). Briefly, extracted DNA samples were quantified using a Qubit 4 fluorometer and Qubit™ dsDNA HS Assay Kit (Thermofisher Scientific). DNA libraries were then prepared using the Nextera XT DNA Library Preparation Kit (Illumina) and the Nextera Index Kit (Illumina) according to the manufacturer's protocol, with minor modifications. Standard protocols were used for 1 ng total DNA input. Genomic DNA was fragmented using a proportional amount of Illumina Nextera XT Fragmentation Enzyme. Combinatorial dual indexes were added to each sample, followed by 12 cycles of PCR to construct libraries. DNA libraries were purified using AMpure magnetic beads (Beckman Coulter), eluted in QIAGEN EB buffer, and quantified using a Qubit 4 fluorometer and Qubit® dsDNA HS Assay Kit. DNA libraries were finally pooled together for sequencing on an Illumina HiSeqX. Raw reads from metagenomic samples were analyzed by CosmosID metagenomics software (CosmosID Inc., Rockville, MD) to reveal strain-level microbial identification in specimens as described elsewhere (Ponnusamy et al., 2016; Hasan et al., 2014; Ottesen et al., 2016). In brief, the system utilizes a high-performance data mining k-mer algorithm and a highly curated dynamic comparator database that rapidly disambiguates millions of short reads into individual genomes or genes that give rise to specific sequences. This read querying allows for sensitive yet highly accurate detection and taxonomic classification of microbial reads.The resulting statistics were analyzed to return fine-grained taxonomic and relative abundance estimates for the microbial dataset.

[0150] Bioinformatics analysis and statistical modeling of the gut microbiome were then performed with the collaboration of INRAe-Metagenopolis (Paris, France).MetagenomicsThe metagenomic species pangenome (MSP) is a repertoire of genes that consists of genes that are present in all strains ("core genes") and genes that are present in only a subset of them ("accessory genes") (Medini, D., et al., 2005, Current opinion in genetics & development, 15(6), pp.589-594.). MSPminer (Plaza Onate, F., et al., 2019, Bioinformatics, 35(9), PP.1544-1552), a clustering tool developed by Plaza Onate et al. (n is n with a tilde), is able to group co-abundant genes into MSPs based on the largest available gene abundance table of human gut microbiota, which contains 10.4 million genes isolated from 1267 fecal samples. MSP abundance profiles were calculated as the average abundance of 100 marker genes defined as the strong basis of each MSP cluster.

[0151] Association of multiple omic data was performed through parallel and vertical integration schemes. Variable selection was performed at the single omic level, and more precisely, univariate statistical analysis (Wilcoxon test) was performed for each omic data set to examine individual molecular level differences between a given phenotype of interest (Aronia vs. control). Parallel integration treated each type of omic measurement equally, and integration identified all single significantly different features simultaneously in a joint model by collecting and correlating them. Spearman correlations were calculated between plasma and urinary metabolite concentrations, clinical outcomes, and intestinal function, and microbial species.

[0152] Research results A total of 323 volunteers visited the unit and were screened to participate in the study, of which 221 were excluded and 102 (47 males, 55 females) were enrolled and randomly assigned to one of the two intervention arms to constitute the intention-to-treat (ITT) population. Two participants discontinued the intervention for personal reasons and three were lost to follow-up, accounting for a total of 5% dropouts. Ninety-seven participants completed all visits and were therefore included in the analysis. Along with these five dropouts, five participants were excluded from the per-protocol (PP) population due to low compliance (<80%, n=1), a history of aortic stenosis (n=1), elevated GGT, ALT and TG at visit 2 with high alcohol consumption at the screening visit (n=1), abnormally elevated cholesterol levels associated with thyroid deficiency (n=1), and suspected influenza at visit 2 (n=1).

[0153] All clinical results are presented in the ITT population. All microbiome analyses were performed on the PP population. In addition, some fecal samples were not analyzed due to antibiotic intake within 3 months (n=4), missing fecal samples (n=2), and failure to pass the quality check based on hierarchical clustering (n=1). The remaining population considered for all microbiome analyses consisted of 85 individuals (Aronia, n=42; control, n=43).

[0154] No statistically significant differences were found at baseline between the control and Aronia groups, except that heart rate during sleep was higher in the Aronia group (p=0.031).

[0155] Analysis of the 7-day food diaries completed by participants at baseline (1 week before the start of the study) showed no significant differences in micronutrients, macronutrients, and PP at baseline between the diets of participants in the Aronia and control groups, except for vitamin B3 (p=0.047), which was higher in the control group (36.2 mg vs. 40.8 mg).

[0156] 24-h, awake, and sleep PWV, central AIx (AIxao), and peripheral AIx (AIxbr) were measured using Arteriograph24™ before visits 1 and 2. Twelve weeks of Aronia extract intake resulted in a significant reduction in 24-h and awake peripheral and central AIx compared to controls (Δ24-h AIxbr=-6.8%, p=0.003; Δ24-h AIxao=-3.3%, p=0.006; Δawake AIxbr=-6.1%, p=0.020; Δawake AIxao=-2.9%, p=0.034). Non-significant trends for Aronia reduction were also observed for sleep AIxao and AIxbr. A significant reduction in awake PWV was also observed with Aronia compared to controls (ΔPWV=-0.24m / s, p<0.05). Similar results were found for the BACO analysis. The results are shown in Table 1.

[0157] [Table 1]

[0158] Values ​​are expressed as mean ± SD. AIx, augmentation index; ao, aortic; br, brachial; CFB, change from baseline; CFC, change from control; PWV, pulse wave velocity. *Significant difference between baseline and week 12 at p < 0.05.

[0159] Table 1 - Effects of Aronia berry extract after 12 weeks of intake on 24-h, awake and asleep central PWV, and peripheral and central AIx in the ITT population and in each intervention group after BAB analysis.

[0160] Consideration The novel findings of this study are the observed significant differences in 24-h and awake AIxao (6.8 and 6.1%, respectively) and 24-h and awake AIxbr (3.3 and 2.9%, respectively) as well as a reduction in awake PWV of 0.24 m / s after 12 weeks of Aronia supplementation compared to placebo. PWV and AIx are gold standard techniques for the assessment of arterial stiffness, assessing both arterial structure and function. Both techniques are known to be strongly correlated with CVD.

[0161] Example 4 - Investigation into the effects of Aronia melaeocarpa extract on the intestinal flora The objective of this study was to investigate the effects of Aronia consumption on the abundance and composition of the gut microbiome and to explore the associations between the gut microbiome, Aronia polyphenol metabolites, and vascular outcomes.

[0162] Fecal samples were collected at baseline and after 12 weeks of daily intake of Aronia extract or placebo and immediately stored at −80° C. Detailed information regarding the collection and processing of fecal samples can be found in Example 2.

[0163] The richness of the samples was assessed at the gene and MSP levels. A significant difference was found in the number of genes between the Aronia and control groups when considering the change from baseline (p=0.021) (Figure 2).

[0164] These results indicate that ingestion of Aronia extract for 12 weeks resulted in an increase in the number of genes, suggesting a favorable increase in the richness of the gut microbiota after Aronia consumption.

[0165] Similar analyses of bacterial abundance and composition were performed for changes from baseline to represent differences in species between the two treatment groups after 12 weeks of daily intake of Aronia extract or placebo (Figure 4). A total of 18 and 4 species were significantly more abundant in the Aronia and control groups, respectively. Among them, Intestinimonas butyriciproducens was the most abundant bacterium in the Aronia group compared to the control group after 12 weeks of intervention. Other bacteria that were significantly more abundant in the Aronia group were: Clostridiales bacterium, Oscillibacter sp., Firmicutes bacterium CAG 103, Lawsonibacter asaccharolyticus, Oscillospirales 5, Clostridium sp., Butyricimonas faecihominis, Turicibacter sanguinis, Bacteroides dorei, Oscillospiraceae, Bacteroides xylanisolvens, Ruminococcus sp. / Blautia sp., Dialister invisus, Flavonifractor sp. / Clostridium sp., Faecalibacterium prausnitzii 2, Christensenellales, and Blautia A.

[0166] The beneficial effects of the treatment on the gut microbiome and its impact on several functionally related pathways (|cliff delta|>0.2) were investigated (Table 2). Thirteen pathways were identified as relevant and associated with the Aronia group, including seven with statistical significance. These include the production of gamma-aminobutyric acid (GABA), a neurotransmitter produced by bacteria such as Lactobacillus to lower intracellular pH; histidine degradation and reduction of pyruvate to ferredoxin, both of which are involved in the formation of the SCFA acetate; gamma-hydroxybutyrate degradation, associated with a pathway leading to the production of butyrate superoxide dismutase, an antioxidant enzyme involved in oxidative stress responses; polysaccharide A, a capsular carbohydrate found in Bacteroides fragilis that presents anti-inflammatory properties, as well as a pathway involved in the production of the SCFA propionic acid. Three pathways were associated with the control group, including two with statistical significance.

[0167] [Table 2] Table 2 - Functional pathways associated with the Aronia and control groups when considering the change from baseline.

[0168] Consideration It is well established that host health is influenced by the composition of the gut microbiome, and the role of diet on microbiota-mediated outcomes. Thus, a reduction in gene richness of the gut microbial ecosystem has already been reported in both rat and human studies (Li et al., 2017) in conditions of gut dysbiosis associated with disease (Le Chatelier, et al., Nature, 500(7464), pp. 541-546), as well as in conditions of prehypertensive and hypertensive conditions. Our analysis shows a beneficial effect exerted by the intake of Aronia extract on the composition of the gut microbiome, as observed as an increased level of gene counts in the Aronia group compared to the control group, which probably contributes to the positive improvement in arterial outcomes observed.

[0169] Furthermore, after 12 weeks of supplementation, several beneficial species were found to be enriched in the Aronia group compared to the control group. Notably, increased levels of the xylene-degrading bacterium B. xylanisolvens were reported, along with species of the butyrate-producing bacteria taxonomy, such as Faecalibacterium prausnitzii (Miquel, S., et al., Gut microbes, 5(2), pp.146-151), Lawsonibacter asaccharolyticus (Sakamoto, M., et al., International journal of systematic and evolutionary microbiology, 68(6), pp.2074-2081.), and the microorganism Intestinimonas butyriciproducens, which is associated with a healthy gut layout. Xylan, as a dietary fiber, is fermented by the human gut microbiota, resulting in the production of short-chain fatty acids. Consistently, these observations were substantiated by the expansion of a potentially functional module leading to the production of propionate, a diet-related gut microbial metabolite shown to play an important role in cardiometabolic health and hypertension (Muralitharan, RR et al., Journal of human hypertension, 35(2), pp.162-169).

[0170] Example 5 - Identification of "responder" subpopulations A high variability in vascular response to Aronia intake was found among the study population. We hypothesized that this interindividual variability in response may be related to differences in the gut microbiome at baseline. Responders (R) vs. non-responders (NR) were classified based on unsupervised clustering k-means among participants from the Aronia group (n=42).

[0171] The following variables were included in the model as relevant clinical parameters: ambulatory blood pressure (BP) (primary outcome), clinical SBP, ambulatory AIx and PWV, FMD and cortisol level. All clinical parameters were primary and secondary outcomes of the RCT, including ambulatory blood pressure (BP) (primary outcome), clinical SBP, ambulatory AIx and PWV, FMD, and cortisol level (Table 3). The cutoffs for each parameter were determined based on clinical relevance, the overall magnitude of effect and range of response in our study, and the ability to perform statistical analysis on large enough groups (Table 3).

[0172] [Table 3] ao, aortic; br, brachial; BP, blood pressure; CVD, cardiovascular disease; DBP, diastolic BP; FMD, flow-mediated dilation; PWV, pulse wave velocity; SBP, systolic BP Table 3: Summary of parameters included in the cluster analysis and details of their cut-off limits.

[0173] The best number of clusters (k=2) was defined using the NbClust() function (NbClust R package) on a matrix containing the changes in the given variables. The analysis was repeated 200 times to ensure the robustness of the observations, and 37 of the 42 volunteers were included in two clusters. The other five participants were classified as "jumping subjects" because they oscillated from one cluster to another during the repetitions.

[0174] Aronia cluster 1 consisted of 23 volunteers, and Aronia cluster 2 consisted of 14 volunteers.

[0175] To evaluate the impact of Aronia treatment in the newly defined subcohort, differences in clinical parameters between the two clusters were investigated. Aronia cluster 1 had a significant reduction in the primary outcome, 24-h SBPbr, by -6.6 mmHg and -3.6 mmHg compared to Aronia cluster 2 and the control group, respectively (p<0.01). Similar observations were found for 24-h DBPbr (reduction of -4.6 mmHg and -2.8 mmHg in cluster 1 compared to cluster 2 and the control, respectively, p<0.01). These observations suggest that subjects in Aronia cluster 1 responded to intake of Aronia extract (they were "responders", R), whereas subjects in Aronia cluster 2 were non-responders (NR) (Figure 4). Furthermore, a significant decrease in awake SBPbr and DBPbr was found in the R group compared with the NR and control groups (Figure 4; -5.3mmHg and -3.8mmHg for awake SBPbr, and -4.6mmHg and -3.3mmHg for awake DBPbr compared with the NR and control groups).

[0176] As shown above, intake of Aronia extract for 12 weeks resulted in an increase in the number of genes, implying a beneficial increase in the richness of the gut microbiota after Aronia intake. Responders had significantly lower gene counts at baseline compared to the NR and placebo groups (Figure 5).

[0177] The same conclusion was observed for species richness, but it was not significant after Bonferroni adjustment for multiple comparisons (Kruskal-Wallis p-value=0.18, Dunn post-hoc p-value (R vs. NR)=0.03, uncorrected). In addition, functional analysis of the gut microbiome highlighted that subjects belonging to the R group had significantly lower functional modules at baseline compared to the NR group (chi-squared p-value for the NR group <2.2e-16), which further confirmed that R individuals have lower richness at baseline levels compared to NR in terms of gut bacterial composition and associated potential functions.

[0178] Baseline gut microbiome composition was investigated for both R and NR subgroups of Aronia subjects. After Kruskal Wallis test with Dunn post-hoc test, the abundance of Faecalibacterium prausnitzii 8, Acutalibacteraceae 3, Firmicutes bacterium CAG 103, and Bifidobacterium adolescentis taxa were considered significantly enriched at baseline in the R group compared to the NR group. Bifidobacterium adolescentis was also enriched in the R group compared to the control group (Figure 6).

[0179] Analyses were conducted focusing only on the four species (listed above) enriched in the R cluster to predict the response to Aronia extract supplementation according to baseline gut microbiota composition variations. We observed that baseline abundance of Bifidobacterium adolescentis was significantly and negatively correlated with elevated 24-h DBP in the Aronia group (Spearman's ρ = -0.32, p = 0.05) (Figure 7).

[0180] Analysis of gut microbial composition based on variation in response to Aronia extract treatment (responders vs. non-responders) revealed that intake of Aronia extract reduced BP (24-h free-moving and awake SBPbr and DBPbr) in subjects with lower gene counts at baseline compared to controls. Of the four taxa significantly enriched at baseline in the R group compared to the NR group, a higher abundance of Bifidobacterium adolescentis was associated with a greater reduction in 24-h DBP.

[0181] The effect of Aronia extract supplementation in the R and NR subgroups was evaluated by comparing the gene counts and composition of the gut microbiome in both subgroups after 12 weeks of intervention.

[0182] Intake of Aronia extract for 12 weeks led to an increase in gene count in the R group compared to both the NR group (p=0.0008) and the control group (p=0.01) (Figure 8). This result indicates that the beneficial effect of Aronia on this parameter was stronger in individuals carrying a lower gene count at baseline.

[0183] Regarding the composition of the gut microbiome, a total of 26 and 10 species were significantly more abundant in the R and NR groups, respectively, after the intervention (Figure 9). Among the species enriched in the R group, the following were found: Gemmiger, Lachnospiraceae G, Intestimonas butyriciproducens, Eggerthella lenta, Lawsonibacter, Faecalibacterium prausnitzii (1, 9, 2, 6), Oscillospirales (3, 5), Faecalibacterium, Roseburia intestinalis, Ruthenibacterium lactatiformans, Lachnoclostridium species, Acutalibacteraceae 3, Clostridium sp., Collinsella bouchesdurhonensis, Firmicutes bacterium CAG 103 / Clostridium sp., Dysomobacter welbionis, Ruminococcus sp., Clostridiales bacterium, and bacteria of the Clostridiaceae family. In contrast, species significantly enriched in NR individuals were Streptococcus salivarius, Oscillospirales 4, Streptococcus australis, Eubacterium species, unclassified Lachnospiraceae C, Firmicutes bacterium CAG 103, Bacteroides vulgatus, Ruminococcus bicirculans, Lachnospira pectinoschiza, and Intestinibacter.

[0184] Several species enriched in the R group are known to be members of the healthy gut microbiome, such as several species of the Faecalibacterium clade, as well as Roseburia intestinalis, a fibrolytic gut commensal bacterium that can affect atherosclerosis in in vivo models (La Rosa et al., 2019), and Eggerthella lenta, a human colonic taxon that produces urolithin metabolites from the metabolism of pomegranate ellagitannins, which has recently been shown in preclinical studies to improve intestinal barrier function and be associated with lower cardiometabolic risk (Selma et al., 2018).

Claims

1. 1. A composition for use in treating and / or preventing gut microbial dysbiosis imbalance in a subject, the composition comprising an extract obtained or obtainable from Aronia melanocarpa.

2. (a) treating and / or preventing hypertension; and / or (b) treating and / or preventing prehypertension, and / or (c) reducing blood pressure; and / or (d) reducing arterial stiffness 10. A composition for use as defined in claim 1, further comprising at least one of:

3. Treating and / or preventing gut dysbiosis imbalance may be achieved by: (a) increasing microbiome diversity as measured by fecal microbiome gene count; and / or (b) increasing the levels of beneficial bacteria such as Faecalibacterium prausnitzii 2, Lawsonibacter asaccharolyticus, Intestinimonas butyriciproducens, Faecalibacterium, Roseburia intestinalis, and / or Eggerthella lenta.

10. A composition for use as defined in claim 1, comprising at least one of:

4. the extract obtained or obtainable from Aronia melanocarpa is an aqueous extract or a hydroalcoholic extract, Optionally, the composition for use according to claim 1 comprises at least 10%, at least 20%, 20%, 40%, at least 50% w / w or more polyphenols by weight of the composition.

5. 2. The composition for use according to claim 1, wherein treating and / or preventing gut dysbiosis imbalance is in a mammal having a fecal microbiome gene count of less than 400,000, optionally less than 375,000, optionally less than 350,000, 300,000, 275,000, 250,000, 225,000, 200,000, 175,000, or less than 150,000.

6. 10. The composition for use of claim 1, wherein the subject is prehypertensive, optionally wherein the subject is receiving: a systolic blood pressure of at least 120 mmHg, optionally at least 130 or at least 140 mmHg; and / or a systolic blood pressure of between 120 mmHg and 130 mmHg, or between 120 mmHg and 140 mmHg, and / or a diastolic blood pressure of at least 80 mmHg, optionally at least 90 mmHg; and / or a diastolic blood pressure of between 80 mmHg and 90 mmHg. The composition having the formula:

7. The dosage for treating gut dysbiosis imbalance is: a) about 100 to about 1000 mg / day by weight of the composition, optionally 100-1000 mg / day, or 200-800 mg / day, or 400-600 mg / day by weight of the composition; b) 1000 mg / day or less, optionally 900 mg / day, 800 mg / day, 700 mg / day, 600 mg / day, 500 mg / day, 400 mg / day, 300 mg / day, 200 mg / day or less, or 100 mg / day or less by weight of the composition; and / or c) 100 mg / day or more, or optionally 200 mg / day or more, 300 mg / day, 400 mg / day, 500 mg / day, 600 mg / day, 700 mg / day, 800 mg / day, 900 mg / day or more, or 1000 mg / day or more The composition for use according to claim 1, which may be:

8. The dosage for treating gut dysbiosis imbalance is: a) from about 100 mg / day to about 500 mg / day of polyphenols by weight of the composition, optionally from about 200 mg / day to about 400 mg / day of polyphenols, optionally about 300 mg / day by weight of the composition; b) no more than 500 mg / day of polyphenols by weight of the composition, optionally no more than 400 mg / day, 300 mg / day, 200 mg / day, or 100 mg / day of polyphenols by weight of the composition; and / or c) 100 mg / day or more, optionally 200 mg / day, 300 mg / day, 400 mg / day or more, or 500 mg / day or more of polyphenols by weight of the composition 2. The composition for use according to claim 1, wherein

9. 4. The composition for use of claim 3, wherein determining the abundance of bacteria in a fecal sample obtained from a subject is performed using shotgun sequencing.

10. 1. A method for determining whether a subject is likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, comprising: a) determining the number of fecal microbiome genes in a fecal sample obtained from the subject; and / or b) determining the abundance of any one, two, three, or all of the following: i) Faecalibacterium prausnitzii 8 in fecal samples obtained from subjects; ii) Acutalibacteraceae 3 in fecal samples obtained from subjects; iii) Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject; and / or iv) Bifidobacterium adolescentis in a fecal sample obtained from a subject Including, Optionally, a) comparing the fecal microbiome gene count in a fecal sample obtained from the subject with the fecal microbiome gene count in at least a first control sample or population of control samples; and / or b) Comparing the following: i) the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from the subject and the abundance of Faecalibacterium prausnitzii 8 in at least a first control sample or a population of control samples; ii) the abundance of Acutalibacteraceae 3 in a fecal sample obtained from the subject and the abundance of Acutalibacteraceae 3 in at least a first control sample or a population of control samples; iii) the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject and the abundance of Firmicutes bacterium CAG 103 in at least a first control sample or a population of control samples; and / or iv) the abundance of Bifidobacterium adolescentis in a fecal sample obtained from the subject and the abundance of Bifidobacterium adolescentis in at least a first control sample or a population of control samples. further comprising Optionally, at least a first control sample or population of control samples is a negative control sample or population of control samples; Optionally, the negative control sample or population of control samples are fecal samples collected from one or more negative control subjects who have been determined to be unresponsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; wherein the sample is taken from one or more control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; The method.

11. 11. The method of claim 10, wherein the fecal microbiome gene count of a sample obtained from a subject is determined by: a) fecal microbiome gene counts in a negative control sample or population of control samples, optionally wherein the negative control sample or population of control samples are fecal samples taken from one or more negative control subjects shown to be unresponsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the samples are taken from the one or more negative control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; and / or b) fecal microbiome gene counts in a positive control sample or population of control samples, optionally wherein the positive control sample or population of control samples are fecal samples taken from one or more positive control subjects shown to be responsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the samples were taken from the one or more positive control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa. The method of claim 1, wherein

12. a) if the fecal microbiome gene count of the sample obtained from the subject is substantially similar to or higher than the fecal microbiome gene count of the negative control sample(s), the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; Optionally, wherein the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa if the fecal microbiome gene count of the sample obtained from the subject is at least 5% greater, such as at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, such as at least 200% greater than the fecal microbiome gene count of the negative control sample(s); and / or b) if the fecal microbiome gene count in the test sample is substantially similar to or lower than the fecal microbiome gene count of the positive control sample(s), the subject is considered likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; Optionally, herein, the subject is considered to be likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa if the fecal microbiome gene count of the sample obtained from the subject is at least 5% lower, e.g., at least 10% higher, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, e.g., at least 200% lower, than the fecal microbiome gene count of the positive control sample(s). The method of claim 10.

13. a) i) comparing the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from the subject with the abundance of Faecalibacterium prausnitzii 8 in at least a first control sample or a population of control samples; ii) comparing the abundance of Acutalibacteraceae 3 in a fecal sample obtained from the subject with the abundance of Acutalibacteraceae 3 in at least a first control sample or a population of control samples; iii) the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject is compared with the abundance of Firmicutes bacterium CAG 103 in at least a first control sample or a population of control samples; and / or iv) comparing the abundance of Bifidobacterium adolescentis in a fecal sample obtained from the subject with the abundance of Bifidobacterium adolescentis in at least a first control sample or a population of control samples; wherein the control sample or population of control samples is a negative control sample or population or control sample; wherein the negative control sample or population of control samples is a fecal sample collected from one or more negative control subjects that have been shown to be unresponsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the samples are collected from one or more positive control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; and / or b) i) comparing the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from the subject with the abundance of Faecalibacterium prausnitzii 8 in at least a first control sample or a population of control samples; ii) comparing the abundance of Acutalibacteraceae 3 in a fecal sample obtained from the subject with the abundance of Acutalibacteraceae 3 in at least a first control sample or a population of control samples; iii) the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject is compared with the abundance of Firmicutes bacterium CAG 103 in at least a first control sample or a population of control samples; and / or iv) comparing the abundance of Bifidobacterium adolescentis in a fecal sample obtained from the subject with the abundance of Bifidobacterium adolescentis in at least a first control sample or a population of control samples; wherein the control sample or population of control samples is a positive control sample or population or control sample; wherein the positive control sample or population of control samples is a fecal sample collected from one or more positive control subjects shown to be responsive to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the samples are collected from the one or more positive control subjects prior to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa. The method of claim 10.

14. a) if the abundance of Faecalibacterium prausnitzii 8 in the sample obtained from the subject is substantially similar to or higher than the abundance of Faecalibacterium prausnitzii 8 in the positive control sample(s), the subject is considered likely to respond to treatment with the composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Faecalibacterium prausnitzii 8 in the sample obtained from the subject is substantially similar to or higher than the abundance of Faecalibacterium prausnitzii 8 in the positive control sample(s). 8 is at least 5% greater, such as at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, such as at least 200% greater, the subject is considered likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; b) if the abundance of Acutalibacteraceae 3 in the sample obtained from the subject is substantially similar to or higher than the abundance of Acutalibacteraceae 3 in the positive control sample(s), the subject is considered likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Acutalibacteraceae 3 in the sample obtained from the subject is at least 5% greater, such as at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, such as at least 200% greater, than the abundance of Acutalibacteraceae 3 in the positive control sample(s); c) if the abundance of Firmicutes bacterium CAG 103 in the sample obtained from the subject is substantially similar to or higher than the abundance of Firmicutes bacterium CAG 103 in the positive control sample(s), the subject is likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Firmicutes bacterium CAG 103 in the sample obtained from the subject is substantially similar to or higher than the abundance of Firmicutes bacterium CAG 103 in the positive control sample(s). the subject is considered likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa if the abundance of Aronia melanocarpa is at least 5% greater, such as at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, such as at least 200% greater than the abundance of 10; and / or d) if the abundance of Bifidobacterium adolescentis in the sample obtained from the subject is substantially similar to or higher than the abundance of Bifidobacterium adolescentis in the positive control sample(s), the subject is considered likely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Bifidobacterium adolescentis in the sample obtained from the subject is at least 5% greater, such as at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% greater, such as at least 200% greater, than the abundance of Bifidobacterium adolescentis in the positive control sample(s); e) if the abundance of Faecalibacterium prausnitzii 8 in the sample obtained from the subject is substantially similar to or lower than the abundance of Faecalibacterium prausnitzii 8 in the negative control sample(s), the subject is considered unlikely to respond to treatment with the composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Faecalibacterium prausnitzii 8 in the sample obtained from the subject is substantially similar to or lower than the abundance of Faecalibacterium prausnitzii 8 in the negative control sample(s). 8 is at least 5% lower, e.g., at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, e.g., at least 200% lower, than the abundance of 8, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; f) if the abundance of Acutalibacteraceae 3 in the sample obtained from the subject is substantially similar to or lower than the abundance of Acutalibacteraceae 3 in the negative control sample(s), the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Acutalibacteraceae 3 in the sample obtained from the subject is at least 5% lower, such as at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, such as at least 200% lower, than the abundance of Acutalibacteraceae 3 in the negative control sample(s); g) if the abundance of Firmicutes bacterium CAG 103 in the sample obtained from the subject is substantially similar to or lower than the abundance of Firmicutes bacterium CAG 103 in the negative control sample(s), the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Firmicutes bacterium CAG 103 in the sample obtained from the subject is substantially similar to or lower than the abundance of Firmicutes bacterium CAG 103 in the negative control sample(s). the abundance of 103 is at least 5% lower, e.g., at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, e.g., at least 200% lower, than 103, the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa; and / or h) if the abundance of Bifidobacterium adolescentis in the sample obtained from the subject is substantially similar to or lower than the abundance of Bifidobacterium adolescentis in the negative control sample(s), the subject is considered unlikely to respond to treatment with a composition comprising an extract obtained or obtainable from Aronia melanocarpa, optionally wherein the abundance of Bifidobacterium adolescentis in the sample obtained from the subject is at least 5% lower, such as at least 10% greater, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, at least 100% lower, such as at least 200% lower, than the abundance of Bifidobacterium adolescentis in the negative control sample(s). The method of claim 10.

15. i) comparing the abundance of Faecalibacterium prausnitzii 8 in a fecal sample obtained from the subject with the abundance of Faecalibacterium prausnitzii 8 in at least a first control sample or a population of control samples; ii) comparing the abundance of Acutalibacteraceae 3 in a fecal sample obtained from the subject with the abundance of Acutalibacteraceae 3 in at least a first control sample or a population of control samples; iii) comparing the abundance of Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject with the abundance of Firmicutes bacterium CAG 103 in at least a first control sample or a population of control samples; and iv) Bifidobacterium adolescentis in a fecal sample obtained from a subject is compared with the abundance of Bifidobacterium adolescentis in at least a first control sample or a population of control samples; The method of claim 10.

16. The abundance of Bifidobacterium adolescentis in fecal samples obtained from subjects is: one or more negative control samples; and / or One or more positive control samples compared with the abundance of Bifidobacterium adolescentis in The method of claim 10.

17. 11. The method of claim 10, wherein the number of fecal microbiome genes in a fecal sample obtained from the subject is less than 400,000, optionally less than 375,000, optionally less than 350,000, 300,000, 275,000, 250,000, 225,000, 200,000, 175,000, or less than 150,000.

18. The number of fecal microbiome genes in a fecal control sample or a population of fecal control samples is determined by shotgun sequencing; and / or in a fecal control sample or a population of fecal control samples, i) Faecalibacterium prausnitzii 8 in fecal samples obtained from subjects; ii) Acutalibacteraceae 3 in fecal samples obtained from subjects; iii) Firmicutes bacterium CAG 103 in a fecal sample obtained from the subject; and / or iv) Bifidobacterium adolescentis in a fecal sample obtained from a subject as determined by shotgun sequencing of fecal control samples. The method of claim 10.

19. 11. The method of claim 10, wherein response to treatment is indicated by a decrease in blood pressure, optionally wherein the method is a method for determining whether a composition comprising an extract obtained or obtainable from Aronia melanocarpa is likely to reduce blood pressure in a particular subject.

20. The subject is prehypertensive, optionally wherein the subject is a patient receiving one of the following: a systolic blood pressure of at least 120 mmHg, optionally at least 130 or at least 140 mmHg; and / or a systolic blood pressure of between 120 mmHg and 130 mmHg, or between 120 mmHg and 140 mmHg, and / or a diastolic blood pressure of at least 80 mmHg, optionally at least 90 mmHg; and / or a diastolic blood pressure of between 80 mmHg and 90 mmHg.

11. The method of claim 10, comprising:

21. 10. A composition for use in treating and / or preventing gut dysbiosis imbalance in a subject, the composition comprising an extract obtained or obtainable from Aronia melanocarpa, wherein the subject has been determined to be likely to respond to treatment with the composition of any one of claims 1 to 8.

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